AICE* SURVEY OF USSR AIR POLLUTION LITERATURE
Volume X!
A SECOND COMPILATION OF TECHNICAL REPORTS
ON THE
BIOLOGICAL EFFECTS AND THE PUBLIC HEALTH ASPECTS
OF ATMOSPHERIC POLLUTANTS
Edited By
M. Y. Nultonson
The material presented here is part of a survey of
USSR literature on air pollution
conducted by the Air Pollution Section
AMERICAN INSTITUTE OF CROP ECOLOGY
This survey is being conducted under GRANT 1 ROl AP00786 - APC
OFFICE OF AIR PROGRAMS
of the
U.S. ENVIRONMENTAL PROTECTION AGENCY
•AMERICAN INSTITUTE OF CROP ECOLOGY
809 DALE DRIVE
SILVER SPRING, MARYLAND 20910
1972
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BIBLIOGRAPHIC DATA
SHEET
1. Report No.
APTD-1067
3. Recipient's Accession Nc
4. Title and Subtitle A.ICE Survey of USSR Air Pollution Literature
Volume XI - A Second Compilation of Technical Reports on the
Biological Effects and the Public Health Aspects of Atnos-
pheric Pollutants
5. Report Date
January 1972
6.
7» Author(s)
M. Y. Nuttonson (Ed.)
8- Performing Organization Rept.
No.
9. Performing Organization Name and Address
American Institute of Crop Ecology
809 Dale Drive
Silver Spring, Maryland 20910
10. Project/Task/Work Unit No.
11. Contract/Grant No.
1 R01 AP00786 APC
12. Sponsoring Organization Name and Address
U. S. ENVIRONMENTAL PROTECTION AGENCY
Office of Air Programs
13. Type of Report & Period
Covered
14.
15. Supplementary Notes
16. Abstracts Ihirteen reports were translated Iron Russian. Seven are rrom;Akademiya Medit-
sinakikh Nauk SSSR. "Biologicheskoe deystvie i gigienicheskoe znachenie atmosfernykh za-
gryazneniy". Red. V.A. Ryazanova. Vypusk 11, Izdatel'stvo "Meditsina" Moskva, (1968)'.
Maximum Permissible Concentrations of Noxious Substances in the Atmospheric Air of Popu-
lated Areas, V.A. Ryazanov, p.201-204; Combined Effect of Hydrogen Fluoride and Sulfur
Dioxide on the Body of Man and Animals, Z. Ya. Lindberg, p.32—43; New Data for the Vali-
dation of the Mean Daily Maximum Permissible Concentration of Hydrogen Fluoride in Atmos
pheric Air, M.S. Sadilova, E.G. Plotko, and L.N. Yel'nichnykh, p.5-15; Sanitary Evalua-
tion of Fluorides Readily Soluble in Biological Media, M.S. Sadilova and E.G. Plotko,
p.16-26; Biological Effect of Poorly Soluble Fluorides, M.S. Sadilova, p.26-32; Reflex
Effect on the Human Organism of Low Concentrations of Acetic Acid and Acetic Anhydride
Present Separately and Together in Atmospheric Air, M.T. Takhirov, p.73-91; Threshold
Concentrations of Paraffins in Short-Term and Long-Term Inhalation, M.L. Krasovitskaya
and L.K. Malyarova, p.43-50. The remaining six translations are from: Akadexiya Medit-
sinskikh Nauk SSSR. 'Biologicheskoe deystvie i gigienicheskoe znachenie atmosfernvkh za-
gryazneniy". Red._ V.A. Ryazanova. Vypusk 10, Izdatel'stvo "Meditsina" Moskva, (19B7):
Basic Proolems of Sanitary Protection of Atmospheric Air, Prof. V.A. Ryazanov, p.5-15;
Material for Standardization of the Maximum Permissible Concentration of Hydrogen Fluor-
ide in the Air of Populated Areas, M.S. Sadilova, p.186-201; Biological Effect and Hy-
gienic Evaluation of Pollution of Atmospheric Air with Phthalic Anhydride, L.P. Slavgor-
orskiy, p.86-96; Data for a Sanitary Assessment of Methanol in Atmospheric Air, R. UBay-
dullayev, p.65-74; Data for the Validation of the Maximum Permissible Concentration of
Ammonia in Atmospheric Air, M.M. Sayfutdinov. p.108-122, (1967); and Pollution of Atmos-
pheric Air with Vapors of Hydrolytic Ethyl Alcohol and its Effect on the Organism, R.
Ubaydullayev, p. 74-86. ,
17. Key Words and Document Analysis. 17a. Descriptors
Air pollution Acetic acid Alcohols Health
Hydrogen fluoride Hydrocarbons , Ammonia
Sulfur dioxide Phthalic acids Ethyl alcohol
17b. identifiers /Open-Ended Terms
Translations
17c. COSATi Field/Group
13B
18. A vailability Statement
Unlimited
19.. Security Class (This
Report)
UNCLASSIFIED
w Securicy Class (This
Page
UNCLASSIFIED
21. No. of Pages
154
22. Price
FORM NTiS-35 (10-701 I USC OMM- D C 4 032 9-P 7 I
/
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TABLE OF CONTENTS
Page
PREFACE v
/'Maps of the USSR
Orientation /. vii
Climatict Soil and Vegetation Zones .. viii
Major Economic 'Areas ix
Major Industrial Centers x
Principal Centers of Ferrous Metallurgy and Main
Iron Ore Deposits xi
Principal Centers of Non-Ferrous Metallurgy and
Distribution of Most Important Deposits of
Non-Ferrous Metal Ores xii
Principal Centers of the Chemical Industry and of
the Textile Industry xiii
Principal Centers of Wood-Working3 Paper, and Food
Industries xiv
Main Mining Centers xv
Principal Electric Potier Stations and Pouer Systems xvi
MAXIMUM PERMISSIBLE CONCENTRATIONS OF NOXIOUS SUBSTANCES IN
THE ATMOSPHERIC AIR OF POPULATED AREAS
V. A. Rvazanov ^ 1
BASIC PROBLEMS OF SANITARY PROTECTION OF ATMOSPHERIC AIR_
V. A. Ryazanov .1^........... 6
COMBINED EFFECT OF HYDROGEN FLUORIDE AND SULFUR DIOXIDE ON
THE BODY OF MAN AND ANIMALS
Z. Ya. Lindberg 14
NEW DATA FOR THE VALIDATION OF THE MEAN DAILY MAXIMUM PERMISSIBLE
CONCENTRATION OF HYDROGEN FLUORIDE IN-ATMOSPHERIC AIRJ ,
M. S. Sadilova, E. G. Plotko, and L. N. Yel'nichnykh 24
SANITARY EVALUATION OF FLUORIDES READILY SOLUBLE IN
RTOLOGICAL MEDIA j '
M. S. Sadilova and E. G. Plotko 33
ill
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Page
BIOLOGICAL EFFECT OF POORLY SOLUBLE FLUORIDES '
M. S. Sadilova . 42
MATERIAL FOR STANDARDIZATION OF THE MAXIMUM PERMISSIBLE
' CONCENTRATION OF HYDROGEN FLUORIDE IN THE-AIR OF,
POPULATED AREAS
M. S. Sadilo.va 48
REFLEX EFFECT ON THE HUMAN ORGANISM OF LOW CONCENTRATIONS OF
ACETIC ACID AND ACETIC ANHYDRIDE:PRESENT SEPARATELY AND
TOGETHER IN ATMOSPHERIC AIR
M. T. Takhirov 61
THRESHOLD CONCENTRATIONS OF PARAFFINS IN SHORT-TESM^AND'
r LONG-TERM. INHALATION
M. L. Krasovicskaya and L. K. Malyarova 78
BIOLOGICAL EFFECT AND HYGIENIC EVALUATION OF POLLUTION OF
ATMOSPHERIC AIR WITH PHTHALIC ANHYDRIDE
L. P. Slavgorodskiy 85
-DATA FOR A SANITARY ASSESSMENT OF METHANOL IN ATMOSPHERIC AIR
R. Ubaydullayev J.... 93
DATA FOR THE VALIDATION OF THE MAXIMUM PERMISSIBLE CONCENTRATION
OF AMMONIAj'-IN ATMOSPHERIC AIR
M M <5oir*utdinov 101
POLLUTION OF-ATMOSPHERI-G-AIR WITH VAPORS OF HYDROLYTIC ETHYL
r ALCOHOL AND ITS EFFECT ON THE ORGANISE
R. Ubaydullayev ; 113
LITERATURE CITED IN 1968 PAPERS 123
LITERATURE CITED IN 1967 PAPERS 128
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PREFACE
The present volume constitutes the second* compilation of technical
reports resulting from a number.of investigations of the biological effects
of specific air pollutants which have been conducted at various public
health institutes and in departments of public health of some of the univer-
sities of the USSR.
The great strides in the development of industrial chemistry in the
country have stimulated studies of the biological effects of chemical air
pollutants as well as studies dealing with public health implications of
these pollutants. Such studies assume an ever-increasing importance in
the Soviet Union.
Professor Ryazanov, in his comprehensive survey "The Basic Problems of
Sanitary Protection of Atmospheric Air", which is presented in this volume,
points out that industrial emissions and the automobile exhausts have become
the primary problem of atmospheric pollution. He suggests that studies of
the biological and toxicological effects of the newly produced chemical
compounds as well as the establishment of their maximum permissible concen-
tration must be intensified and; accelerated as much as possible.
The material included in this volume deals with the biological effect
of low concentrations of chemical toxic substances
(1) emitted from oil refineries,
(2) contained in the discharges of aluminum and superphosphate plants,
(3) contained in the discharges of the industries involved in the
hydrolytic cleavage of wood, and
(4) contained in the emissions from a number of new substances which
are either already employed in the current industrial use or are still
under tests for use in industry.
The results of the above studies provide in the USSR a basis for the
establishment of a series of new maximum permissible concentrations for new
toxic substances in the atmospheric air and constitute the scientific criteria
for assessing the degree of pollution of the air medium. They also form the
foundation for a number of ameliorative sanitation measures to be undertaken.
Some background information on the distribution of the Soviet industry's
production machine may be of interest in connection with that country's present
• Ir.e first compilation of this nature hes been published ir. VoIubh'VIII of ^he AICE Survey of
USSR 4ir Foliation Literature.
V
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and potential pollution problems and investigations. The planned distri-
bution of production in the Soviet Union favors effective exploitation
of the natural resources of the USSR, especially in its eastern areas where
enormous natural resources are concentrated, and has led to the creation
of large industrial centers and complexes of heavy industry in many of
the country's economic areas (see page ix). The many diverse climatic con-
ditions of the country and its major economic areas as well as the geograph-
ical distribution of the Soviet Union's principal industrial and mining
centers and of its principal electric power stations and power systems can
be seen from the various maps presented as background material in this volume.
It is hoped that the papers selected for presentation in this volume
will be conducive to a better appreciation of some of the air pollution
investigations conducted in the USSR. As the editor of this volume I wish
to thank my co-workers in the Air Pollution Section of the Institute for
their valuable assistance.
M. Y. Nuttonson
January 1972
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U.S.S.FL
ORIENTATION M
— Internafonal Boundary
160 • r?o
'F- ~ xS
— Intentional Boundary
— — ¦— Union Republic (SSR)
Aulonomoin Republic (ASSR>
O C *¦ * *N
j
Tundra
¦: *V V*\ ytv /7*»S X\ - /'
K Taiga /
/
V WESTet ) .•?-]/ \
'Krasnoyarsk
/I;
/<¦ I x
rJ u. i
V.-0l»nU,ae ^
\ ,-¦'
Mountains
' **/ **h
+*&
MONGOLIA
Middle
7 AFCANISTAN
CHINA
ADMINISTRATIVE DIVISIONS
SSR
1. RSFSR
2 Kjreto-Finnith SSR
3. Estonian S.S R
4. UtvianSSR
5 Lithuanian S S R
6 While Ruwsian SSR
7 Ukrainian S. S N
8 Moldavian S S R.
9 Georgian SSR
Aimenian SSR
Ajertoydihan $ $.ft
Kazakh SSR
Uttefc S S R
Turkmen S S.R
15 Tadihih SSR
16 KirfiiSSR
A.S.S.M.
A KomiASSH
8 Udmurtskaya ASSR
C Miripkiyi ASSR
0 Chuvattakaya ASSR
L Mofdovtkjya ASSR
f Tatar^kaya ASSR
G Bethkir?kaya ASSR
H Dafestantfcayt ASSR
J Seve/rvOwitinflutya ASSR
K KabardinOmya ASSR
L Abkhuskjya ASSR
M. Adrterskaya ASSR
N Nakhichevanskaya ASSR
0 Kara Kalpaklkaya ASSR
P BuryatMongtf'ikaya ASSR
Q Yakutskaya ASSR
-------
CLIMATIC ZONES AND REGIONS* OF THE USSR
X ARCTIC OCEM? '.
=n
i
i c-Y
<-v.
> * vV"- \ i s*^T~r< \
' 'v^SS^0/* v^./s V?
I
^ "\ \S7>v'/ A^i
?^SEAjOF OKHOTSK
* X^&T ' ^ \
j 'S\
I/-X,/
CJ
Zones: I-arctic, Il-subarctic, IH-temperate , IV-subtropical
Regions: 1-polar, 2-Atlantic, 3-East Siberian, 4-Pacific, 5-Atlantic,
6-Siberian, 7-Pacific, 8-Atlantic-arctic, 9-Atlantic-continental forests,
10-continental forests West Siberian, 11-continental forests East Siberian,
12-monsoon forests, 13-Pacific forests, lA-Atlantic-continental steppe,
15-continental steppe West Siberian, 16-mountainous Altay and Sayan,
17-ir.ountainous Northern Caucasus, 18-continental desert Central Asian,
19-mountainous Tyan-Shan, 20-western Transcaucasian, 21-eastern Transcau-
casian, 22-mountainous Transcaucasian highlands, 23-desert south-Turanian,
24-mountainous Pamir-Alay
(After 3. P. Alisov, "Clisate of The USSR", Moscow 19i>6)
SOIL AND VEGETATION ZONES IN THE U.S.S.R.
.ox ¦: ¦«.
i! 1.1! is1: , (I
¦ fa>nGyar$w' I
fter A, Levrishchev, "Economic Geography
:f the U.S.S.R.", Moscow I960)
viii
-------
MAJOR ECONOMIC AREAS OF THE U.S.S.R.
*,ki j-ubiniV
I Noilh-Weiletn VI Volga XI Bailie
II c«n(i«l Vi: Urali x:i Sojlh-W«
III Ce-if-al Cncmoiem V;il W«,l S.bn« XIII Oor.eli-D
IV Volga-Vyalka IX Eail S'berian XIV Sou!h«m
V North Caucasian X Far Fallen XV Tismcauc
XVI Kaiakhtlart
XVII Cen'fa* Ajior-
XV::i Byclorjjsian
PLANNED DISTRIBUTION OF INDUSTRIAL PRODCCrlCN IN ORDER
IC BRINC IT CLOSER TO RAW MATERIAL AND FCSL SPACES
An example cf the planned distribution of industrial production ir. the L'SSR is the creation of large
industrial centers ar.d complexes of heavy industry in cany of the country's economic areas: the Norch-flest
(Kirevsk, Kandalaksha, Vorkuta), the Urals (Magnitogorsk, Chelyabinsk, Nizhny Tagil), Western and Eastern
Siberia (Novosibirsk, Novokuznetsk, Kemerovo, Krasnoyarsk, Irkutsk, Hratsk), Kazakhstan (Karaganda, Rudnv,
Balkhasn, Dzhezkazgan).
Large industrial systems are being created - Kastanai, Pavlodar-Ekibastuz, Achinsk-Krasnoyarsk,
3ratsk-Taisnet and c number of others. Ferrcus and non-ferrous metallurgy, pulp ar.d paper, hydrolysis ar.d
saw-niiling industries are being established in the Bratsk-Taishet industrial system. The Acr.insk-Kras-
ncyarsk industrial syster. is becoming or.e of the largest centers cf alu-inum ar.d chemical industries, ar.d
production of ferrous metals, cellulose, paper, and oil products.
Construction of the third netallurgical case nas beer, launched ir. Siberia, ar.d a new base of ferrous
metallurgy, using the encr-cus local iron and coal resources, has been created in Kazakhstan. A high-
capacity power syster. is being organized in trio same areas. Non-ferrous metallurgy is being further
developed in Kazakhstan, Central Asia and in Transbaikai areas. The pulp and paper, as %ell as the timber,
industries are being developed st a fast rate in the forest areas cf Siberia ar.d the Far East.
Ferrous metallurgy is also developing in the Europecr part cf the country by-utilizing the enormous
iron ore resources of the Kursk Magnetic- Anosaly and the Ukrainian deposits. Large new production systems
nre under construction in the North-West, along the Volga, in the Northern Caucasus and the Ukraine.
(After A. Lavrisfcchev, "Economic Geography
of the U.S.S.R.", Moscow 1969)
ix
-------
THE MAJOR INDUSTRIAL CENTERS OF THE USSR
^^JyMiiukoQ Uen'"g™d
^'Arkhangelsk
~ MOSCOW.
O'Mage
ostov-on-Don
Yakutsk
o** Kuibyshev s\
o
verdk>v
-------
PRTMPTPAT. PFNTTTPC
V KFRROIK MPTA1 T TTPfTV
J ^Oler^gofsk
"i
Vorkuta
Korihunov
s (i
ftP«l'Oviic
Lgifing?'
5^ Kolpino WK^s" rf AK1'£«S.,oy /
•.. ,-vv lepa'\ Chusovii^^'hny Ta(i1
¦••'¦ ¦ " \_-T lekIFos!a! , 7 0®pAl«pdj£v
moscoww /0-Ji!levskOry va^^5Sv»'Tov'^
OorkyAe'^>u,al,k )
. . — ZfalouslCX ©Chslyabrnik
/ UkkK ^1. ¦_ . t \ByBlorefsj<®XM^nttogorsk
fKudny
CherepoveU
Krlinoyarik
,l./\ v.- • i .
UPo!uoocK>oy«
OVOliblrs
NOVOK
TaiMagol : •;
>
r . / kma.
u^epfo * A ,—
-^"V ? Kramalorsk
, , u-uon«'A
K - K.on\lan ii flov « n
M 'M^Uvitli
mat
Lipefsk
T«rnvl«u
Novolroihk
¦Karaganda
AAlaiu
WMargarels
:%£
;g^®
:.;"K/'iv6i Rog
Aktyubinsk
aganrog
Zaporpt
talon
3ckabadQ
^ Co-rp1«!e cycl« m«l«liur9f
Stvel imc'Hng ard n-elal
if »ollwg
© Sm«Hing of letroa'loys
Mining e(
A *r>s ore*
cok!-g eo*l
ki manghAete ef»'s
"efrc vtk - Zahoi*al$V
Dash
Koirfomolik
or-Amj
IfAIK IRON ORE DEPOSITS IN THE U.S.S.H
^»ui»0901^
^.(Torshuno'3
i -V
V///W'
xi
(After A. iavrishchov, "Eccnamic Geography of
the U.S.S.R.", Moscow 1969)
-------
PRINCIPAL CENTERS OF NON-FERROUS ''METALLURGY IN THE U.S.S.R.
DISTRIBUTION OF MOST IMPORTANT DEPOSITS OF 'TON-FERROUS METAL OPES
C CO'd
Pi P^alinun
Cooper ores
O Tin ores
{^) Complex ores
xii
(After A. Lavrishchev, "Econsr.ic Geogrspr.;,
of the I.'.-S.S.R.", Moscow 1969)
-------
PRINCIPAL CENTERS OF THE CHEMICAL INDUSTRY IN THE U.S.S.R.
^ r w. j. i C
° • V- \ /"N* » ) ^ '
t.i\
• G^c^-afc'v4-''
• vv c.fv
>-pt> \
o »a. "u
-T .r)
x Jk j> c h ^ ^
t ©£ ^ '*
v%
i SS:<^Sf,k.
c«I inc
d -nui.i.-y
O P'cdui'icn of i y r> I "i e I - - r.bbe'
O Prcducl:QP cf minctfl feHili
PRINCIPAL CENTERS OF THE TEXTILE INDUSTRY IN THE UTS.S.R
JC^-KJWC ,_/Jl y'
^QK i-v«h , (Q'
V.1 - .
JPB Z jyr*e, 0
?Oiid (£J>M
'\§ /
\ Ycacyevsk f
thhf^ - C"A
xiii
(/.fear A. L";v»-'r,rol.?v, "Soonomio Geography
ci tne I'.S.S.B.", Mcsiow 19&9)
-------
PRINCIPAL CENTERS OF WOOD-WORKING AND PAPER INDUSTRIES IN THE U.S.S.R.
KOVCS
Kai'rvng
ii ®Odiiss» m5sC
©Kharkov.:
Dnepropei'oy,
ner
Astrakhan
oronais
kom«3iii
Sk#.,'i ¦'
ovqubrtkrfltfV .$fL Cllf^MB.'e'skf- ...1: ••\--:sY'./:• \ -»sa' i£*v/»vi,--ik"r^-i* -
\ .'. ; L : "^KerlM ¦ r- £ ' • -Vv/,^9 ^ akFa irs<
man
aivosfa
T^^hkr il
r.au^ry:
Timber.saw! ig and wood-working
© Paper
C ? Pr nc'pal lumbering areas
Fores's
¦¦mw&y'
IDC3 km
£ Food hdyilry
Q Ffo^r-milling indui'ry
<2) A»e*t hd^jlry
Q Fish arid |iih packing indjjlry
(After A. Lavrishcbev, "Ecor.onic Gcccrap.iy
of -.h« U.S.S.R.", Moscow 19^9/
xiv
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THE MAIN MINING CENTERS OF THE USSR
^Arkhangelsk
Yakutsk
.rasnoyd'sl
habarovsk-
^—O^^-lrkuhi
araganda^M
•Tashkent
MINING
Oil refining
(After A. Efimov, "Soviet Industry", Moscow 1968)
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PRINCIPAL ELECTRIC POWER STATIONS A.ND POI-7ER SYSTEMS IN THE U.S.S.R.
""
Ulorwsfcaya —jr—A/
Ntvjkiyc
Kirovfk
. . . KurnskayA*,
khangenk
-sflufintVh
jherepove!
Vorkuta
*
UkMa
*
North-Weil
and Wesl
.Norilsk
*
Caucasus
Khanlaijkaya
Al-ower Tungujka
Cneboks
Upper Kama
payTovik-K
thelskay
Yakutsk
*•20%?
Bcloyarsky
MukhluyaJ^J
Ounovskaya
Iriktin*
Astrakha
r
Chulman ^
KofnsomoHk
Boguchany
Mamakan
Guryev
aknal
Tseunogra
Khabarovsk
drnalil
—fj K ra snovodsk
* *
Nebit Dag
Irkutsk
Kamenogors
ukhtarmirifkava^• *
•' s^h «ba
"lv*l,CVIidiv5*tok
lzA=f=A:N
Na^o
.•••.v.-::::-:-;::?
::\*rH+r .. ..
. • .{jos
Golpvnaya
Principal Electric Power Stations
Thermal Hydro-power
J^l xjt in operation
A iSl under construction
" " and planned
Groups ol eleclric
power stations
Operating atomic electric power stations
Areas of operation ol single power grids
European part of the U.S.S.R.
Central Siberia
Areas of operation of integrated power grids
Northern Kazakhstan
Central Asia
, „ Figures indicate following power stations:
' Baltic 9 Dnieprodior/hinsk 17 Shatura
2 Narva lOOnieproges 18 Elektrogorsk
3 Kegum tl Kakhovka 19 tvankovo
4 Plavinas 1? Starobeshevsk 20 The 22nd C.P.S.U. Congress
5 Novaya Byetorusskaya 13 Zuyevskaya HEPS on the Volga
6 Dubossary 14 Shterovka 21 The Lenin HEPS on the Yolga
7 Kanev 15 Krasnodar 22 Chardarinskaya
8 Kremenchug 16 Kashira 23 Chirchik-Borsu
24 Nurek
25 Ragunskaya
26 Varzob
?7 Toklogul
29 Alamedi
3fr0 ]
360 km
(After A. Lavrishchev, "Economic Geography
of the U.S.S.R,", Moscow' 19&))
-------
MAXIMUM PERMISSIBLE CONCENTRATIONS OF NOXIOUS SUBSTANCES IN THE
ATMOSPHERIC AIR OF POPULATED AREAS*
*(V. A. Ryazanov)
From Akademiya Meditsiriakikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova,
Vypusk 11, Izdatel'stvo "Meditsina" Moskva, p. 201-204, (1968).'
Pollutant
Concentration, mg/m^
Maximum single
Mean daily
1.
Nitrogen dioxide
0.085
2.
Nitric acid (based on HNO3 molecule
0.4
(based on hydrogen ion)
0,006
3.
Acrolein
0,30
4.
Alpha-methylstyrene
0.04
5.
Alpha-naphthoquinone
0.005
6.
Amyl acetate
0.10
7.
Amylene
1.5
8.
Ammonia
0,20
9.
Aniline
0,05
10.
Acetaldehyde
0,01
11.
Acetone
0,35
12.
Acetophenone
0.003
13.
Benzene
1.5
14.
Gasoline (low-sulfur petroleum
gasoline in terms of "C")
5,0
15.
Shale gasoline (in terms of "C")
0,05
16.
Butane
200.0
17.
Butyl acetate
0.10
18.
Butylene
3,0
19.
Butyl alcohol
0.3
20.
Butyl phosphate
0.01
21.
Valeric acid
0.03
22.
Vanadium pentoxide
0,085
0.4
0.006
0.10
0.04
0.005
0,10
1.5
0.20
0.03
0.35
0,003
0.8
1,5
0,05
0.10
3.0
0,01
0,002
Approved by the Assistant Chief Public Health Physician of the USSH on 12 September 1967* 692-6/
- 1 -
-------
23. Vinyl acetate
24. Hexamethylenediamine
25. Bivinyl
26. Diketene
27. Dimethylaniline
28. Dimethyl sulfide
29. Dimethyl disulfide
30. Dimethylformamide
31. Dowtherm
32. Dichloroethane
33. 2,3-Dichloro-l,4-naphthoquinone
34. Diethylamine
35. Ieopropylbenzene
36. Iaopropylbenzene hydroperoxide
37. Caprolactam (vapors, aerosol)
38. Caproic acid
39. Malathion
40. Xylene
41. Maleic anhydride (vapors, aerosol)
42. Manganese and its compounds (in
terms of Mn02)
43. Butyric acid
44. Mesidine
45. Methanol
46. Metaphos
47. Metachlorophenyl isocyanate
48. Methyl aerylate
49. Methyl acetate
50. Methyl mercaptan
51. Methyl methracylate
52. Monomethylaniline
53. Arsenic (inorganic compounds other
than arsine, in terms of AS)
54. Nitrobenzene
55. Parachloroaniline
56. Parachlorophenyl isocyanate
57. Pentane
58. Pyridine
59. Propylene
60. Propyl alcohol
61. Nontoxic dust
62. Metallic mercury
0.20
0.001
3.0
0.007
0.0055
0,08
0.7
0,03
0.01
3.0
0.05
0.05
0.014
0.007
0.06
0.01
0. 015
0.2
0,2
0. 015
0.003
1.0
0.008
0,005
0. 01
0.07
9-10"6
0.1
0.04
0. 008
0, 0015
100. 0
0.20
0.001
1.0
0.007
0.005
0.2
0.05
0,01
0. 01
0.5
0. 005
0. 003
0.008
0.0015
0. 15
0. 0003
- 2 -
-------
1
2
3
63.
Soot (carbon black)
0.15
0.05
64.
Lead and its compounds (other than
tetraethyl lead) in terms of Pb
0.0007
65.
Lead sulfide
0.0017
66.
Sulfuric acid (based on H2SO4 molecule)
0.3
0.3
(based on hydrogen ion)
0.006
0.006
67.
Sulfur dioxide
0.5
0.05
68.
Hydrogen sulfide
0.008
0.008
69.
Carbon disulfide
0.03
0.01
70.
Hydrochloric acid (based on HCl molecule
) 0.2
0.2
(based on hydrogen ion)
0, 006
0.006
71.
Styrene
0.003
0,003
72.
Thiophene
0-6
73.
Toluylene diisocyanate
0.05
0.02
74.
Toluene
0.6
0.6
75.
Trichloroethylene
4 ". 0
1.0
76.
Carbon monoxide
3-0
1.0
77.
Acetic acid
0.2
78.
Acetic anhydride
0.1
79.
Phenol
0. 01
0. 01
80.
Formaldehyde
0. 035
0. 012
81.
Phosphoric anhydride
0. 15
0. 05
82.
Phthalic anhydride (vapors, aerosol)
0.10
83.
Fluorine compounds (in terms F)
Gaseous compounds (HF, SiF4)
0. 02
0. 005
Soluble inorganic fluorides (NaF,
Na2SiF6)
a 03
0. 01
Sparingly soluble inorganic fluorides
(AlF3, Na3AlF3, CaF2>
0.2
0,03
In the combined presence of gaseous
fluorine and fluorine salts
0.03
0,01
84.
Furfural
0.05
0.05
85.
Chlorine
0.10
0.03
86.
Chlorobenzene
0.10
0.10
87.
Chloropropene
0.10
0.10
88.
Hexavalent chromium (in terms of CrC>3)
0.0015
0.0015
89.
Cyclohexanol
0.06
0.06
90.
Cyclohexanone
0.04
0.04
91.
Carbon tetrachloride
4.0
92.
Epichlorhydrin
0.2
0,2
93.
Ethanol
5.0
5.0
94.
Ethyl acetate
0.1
0.1
- 3 -
-------
95. Ethylene
96. Ethylene oxide
3.0
0.3
3.0
0.03
REMARKS
1. In the combined presence in atmospheric air of several substances
possessing a summation effect, the sum of their concentrations as calculated
by the formula below (§ 2) should not exceed 1 for:
a) acetone and phenol
b) Sulfur dioxide and phenol
c) sulfur dioxide and nitrogen dioxide
d) sulfur dioxide and hydrogen fluoride
e) sulfur dioxide and sulfuric acid aerosol
f) hydrogen sulfide and dowtherm
g) isopropylbenzene and isopropylbenzene hydroperoxide
h) furfural, methanol and ethanol
i) strong mineral acids (sulfuric, hydrochloric and nitric) in
terms of the hydrogen ion concentration (H)
j) ethylene, propylene, butylene and aniylene
should not exceed 1.3 for:
a) acetic acid and acetic anhydride
should not exceed 1.5 for:
a) acetone and acetophenone
b) benzene and acetophenone
c) phenol and acetophenone
2.
Formula for the calculation:
'~X
a
m,
+
1
m5 »>i.
where X is the unknown total concentration:
~ ~ is the concentration of the substance being determined,
-j 1- divided by the corresponding maximum permissible con-
rn' centratlon for isolated action.
a
m,
3. In the combined presence in atmospheric air of:
a) hydrogen sulfide and carbon disulfide
b) carbon monoxide and sulfur dioxide
c) phthalic and maleic anhydrides and alpha-naphthoquinone, the
maximum permissible concentrations for each of them individually
are retained.
-'4 -
-------
4. In the combined presence in atmospheric air of parachlorophenyl
isocyanate and metachlorophenyl isocyanate, temporarily^ until a method of
their isolated determination is developed, the standardization should be
made on the more toxic substance, i. e., parachlorophenyl isocyanate.
5. The maximum permissible concentrations of noxious substances in
the atmospheric air of populated areas as.formulated in December 1966
(No. 655-66), should be considered obsolete.
- 5 -
-------
BASIC PROBLEMS OF SANITARY PROTECTION OF ATMOSPHERIC AIR
Prof. V. A. Ryazanov
From Akademiya Meditsinskikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 10, Izdatel'stvo "Meditsina" Moskva, p. 5-15. (1967),
The problem of atmospheric air pollution, which became very acute about
100 years ago, went through its major historic phase in the last century.
At the present time, the state of the atmosphere has changed so much, and
the sources of its pollution have become so complex, that the very problems
that must be dealt with have become completely different and immeasurably more
complex.
We cannot subscribe to the point of view of our foreign colleagues who
consider the problem of purity of urban air with great pessimism and point to
the inevitability of an increasing atmospheric pollution and the necessity
for mankind to "get used to" or "adapt" to this "disease of civilization,"
since in their opinion nothing else can be done.
Many years of observations and reflection on this problem lead to the
opposite conclusion. However, the striking changes which have occurred in
this problem as a whole should be taken into consideration in order to arrive
at a correct evaluation of the prospects and lines of its development.
Whereas the last century and the first half of the 20th century were
typically characterized by atmospheric pollution caused by the use of mineral
fuel in home heating stoves, fireplaces, chimneys, boilers, and electric
power plants, and the chief components of this pollution - sulfur oxides,
soot, and ash - were the most widely distributed and dangerous, this situa-
tion began to change in the middle of the 20th century. In many countries,
particularly in the USSR, radical steps were initiated to eliminate the
smoke pollution of the atmosphere: home heating stoves and small boilers
were replaced by central heat supplied by heat and electric power plants,
the process of coal combustion was improved so as to raise the efficiency of
the furnaces and decrease the unburnned component, ash collectors with a higfi
degree of purification were constructed, the sulfur content of the fuel was
restricted, and finally, the mineral fuel was replaced by natural gas at
the major heat and electric power plants as well as in consumer use. In
addition, technological and utility processes were electrified with substi-
tution of smoking fuel by electric power. The problem of "smoke" properly
speaking had been solved theoretically by the middle of the 20th century and
also in practical applications in many cities of economically advanced
countries.
- 6 -
-------
There is no doubt that for young developing countriesno other means
of solving the problem of "smoke" can be invented than the one that has
been successful in economically more developed countries (introduction of
a district heating system, installation of gas, electrification). We are
skipping a number of secondary measures whose introduction plays.an auxil-
iary part in the solution of this problem.
In addition to the fuel combustion process, other processes had a
definite but more limited significance in air pollution. For example, the
metallurgical industry contains major sources of polluted air, but their
influence is confined to certain areas. Atmospheric air is frequently
polluted in a given metallurgical district, but outside the limits of the
latter pollution is absent. Dangerous concentrations of atmospheric pol-
lution were produced by nonferrous metallurgical plants. When toward the
end of the 19th century the oxide deposits were exhausted, the industry had
to switch to poor polysulfide ores. This led to a very heavy pollution of
atmospheric air with sulfur dioxide, causing the destruction of green vege-
tation extending many miles outward from the nonferrous metallurgical plants.
Polysulfide ores produced a dust of very complex composition: it included
numerous metallurgical compounds, many of which had a high toxicity. The
pyrometallurgical process involved the conversion of all these extraneous
metallic elements into fumes, which in turn increased the dispersity and
hence the danger of the dust, and the oxidation of these elements with the
formation of oxides was usually accompanied by an increase in their toxicity.
The development of metallurgy of light metals (aluminum, magnesium,
beryllium) was associated with the emission of specific pollutants such as
fluoride compounds, which resulted in the appearance of fluorosis among the
juvenile population, and with the formation of tars containing enormous
amounts of 3,4-benzpyrene and beryllium, which produced an extremely dangerous
disease, berylliosis, among the surrounding population.
However, all these grave calamities suffered by the population as a
result of the development of the metallurgical industry continued to remain
a local phenomenon of regional importance, whereas the smoke produced by the
combustion of fuel constituted a national calamity.
It is completely understandable, therefore, that although the solution to
the control of smoke is still largely theoretical, it is nevertheless of major
and fundamental importance-
In a scientific-theoretical sense, the enterprises of the metallurgical
industry as sources of atmospheric pollution also constitute a practical area
that is slowly yielding to the pressure of science. Theoretically, this
problem has been largely solved. The solution found in this case consists in
the complex utilization of the raw material. Instead of the emission into the
atmosphere of enormous amounts of sulfur dioxide formed in the course of smelt-
- 7 -
-------
ing of metals, these concentrated gases, completely suitable for production of
sulfuric acid, should be regarded as the raw material for sulfuric acid plants,
which should be built with nonferrous metallurgy as their base. Instead of
using polymetal1ic ore, the latter should first be subjected to a fine selective
separation by flotation, and each fraction should be sintered and melted in
separate metallurgical furnaces. The dust residue should be trapped in modern
cloth and fiberglass filters. A metallurgical center should consist of a
complex of plants or shops producing dozens of different products and utilizing
without residue everything of value contained in the ore.
Moreover, it should be noted that nonferrous metallurgy now has a powerful
competitor in the form of plastics, which are rapidly displacing nonferrous
metallurgical products from the consumer market because of their low cost, ease
of fabrication, strength, anticorrosive properties, and a number of other quali-
ties which make plastics irreplaceable, even though many of them are initially
made as substitutes. The age of nonferrous metals is being superseded before
our eyes by the age of plastics, to which the future belongs.
By the end of the first half of the 20th century, in addition to the fumes
resulting from fuel combustion, an increasingly important role in the smoke
pollution of urban atmospheric air had begun to be assumed by the exhaust gases
of motor vehicles. For example in the U.S.A., during the period from 1920 to
1950, i.e., in only 30 years, the number of automobiles increased fivefold,
from 10 to more than 50 million. Motor transport is one of the sources of
urban pollution of atmospheric air with various noxious and offensive fumes,
carbon monoxide, hydrocarbons, aldehydes, and tar compounds, including
carcinogens, products of decomposition of antiknock agents containing a finely
divided smoke of lead oxides, etc. However, the middle of the 20th century
is best characterized by the appearance of the photochemical fog, an entirely
new factor in man's environment.
The photochemical fog, which was incorrectly named "smog", was first
noticed in Los Angeles in 1940. By 1945, the Los Angeles smog had become a
serious problem.
A characteristic feature of the photochemical fog is its formation as a re-
sult of photochemical reactions taking place in the open air in the presence of
sunlight. These photochemical reactions involve organic substances emitted with
the exhaust gases of automobile engines. The most important role in these
processes is thought to be played by olefins, particularly pentene and hexene.
An important part is also attributed to nitrogen oxides, emitted from the exhaust
pipes of engines. A ring reaction takes place with the formation of ozone.
In the presence of light and hydrocarbons, nitrogen dioxide is reconverted into
nitric oxide, and the reaction proceeds as long as sunlight is present. Ozone
in turn reacts with the olefins.
As a result of complex and diverse reactions resulting in the formation of
free radicals, ozonides, and various peroxides, a variety of organic substances
- 8 -
-------
are formed in the atmosphere which differ considerably in chemical activity..
Irritation of the mucous membranes of the eye associated with lachrymation,
damage to vegetation, corrosion of rubber products, and a decrease of visi-
bility are observed. The main complaints of the population concern the
lachrymating effect of the photochemical fog. Prolonged efforts to determine
precisely what specific compound is responsible for the phenomenon of the
photochemical fog have been unsuccessful thus far. It is possible that each
of its effects is due to a different chemical agent. A new compound named
peroxyacetyl nitrate has been successfully extracted from atmospheric air.
It displays the same effects as the photochemical fog, but has not yet been
proven to be the active agent of the Los Angeles smog. In any event, it has
now been established that the photochemical fog is formed as a result of pol-
lution of atmospheric air with exhaust gases in the presence of sufficient
insolation and temperature inversion.
The appearance of the photochemical fog in Los Angeles for the first tine
was attributed to the presence of specific conditions: the lack of public
urban transportation and the consequent excessive saturation of the city with
private automobiles whose number is measured in the millions, the tendency
of this entire region toward anticycIonic weather, and the abundance and
constancy of solar radiation. Wherever such conditions exist, such adverse
consequences are possible in various degrees. In the last few years, in addi-
tion to Los Angeles, the photochemical fog has begun to appear in many other
cities of the U.S.A., including San Francisco, Washington, New York, etc.
The areas covered by the photochemical fog expand each year, and the number of
cities affected by it is growing. . The photochemical fog is becoming the most
"modern" disease of American cities.
It would be difficult to assume that the harmful influence of the photo-
chemical fog is limited solely to the lachrymating effect, although the latter
alone is enough to make life miserable for the entire population (in Los Angeles,
a fog of this type is observed for up to 100 days per year). This is most
probably associated with chronic illnesses that thus far have not been success-
fully identified.
The expanding use of automobiles throughout the world is causing the ad-
verse effects of exhaust gases to increase steadily; phenomena analogous to
the photochemical fog may also arise in other countries. In any case, the
time has come to tackle this problem in the Soviet Union as well.
The A. N. Sysin Institute of General and Communal Hygiene of the USSR
Academy of Madical Sciences has begun some exploratory studies along these
lines. The chemist V.A. Popov has adopted the pheolphthalein method used in
the United States for determining the so-called oxidants in atmospheric air.
Tentative studies in Moscow, Baku and Batumi have shown that in the summertime,
i.e., in the presence of sunlight, oxidants, which are products of photochemi-
cal conversions of exhaust gases, may be detected in the air of these cities.
_ 9 _
-------
Although their concentrations are very slight (approximately eight times
lower than in American cities), they nevertheless approach the limit per-
missible by the California standard. As the number of automobiles grows,
the concentrations of oxidants will rise, and if no decisive steps are taken
to reduce the emission of exhaust gases, it will be impossible to avoid the
appearance of the disease of American cities in the Soviet Union as well.
The universal presence of motor transport and its volume, the steady
growth of the number of automobiles, emission of noxious substances in the
zone where people breathe, ability of the components of these gases to under-
go various photochemical conversions with the participation of highly reactive
free radicals - all of these factors make the pollution of atmospheric air
with automobile emissions the most threatening factor from the standpoint of
modern sanitation. It is also necessary to consider that the exhaust gases
contain many other ingredients such as aerosols of lead compounds, carbon
monoxide, and carcinogens.
It may be stated therefore that at the present stage, the urgency of
this problem has supplanted the importance earlier justifiably accorded by
hygienists throughout the world to the combustion products of mineral fuel.
This problem should become one of the first priorities in our scientific
and environmental efforts.
However, the means of solving this problem do exist. Suffice it to
mention the coordination of intracity transportation, checking of the oper-
ating condition of cars, the removal of transit transport beyond the city
limits, conversion of automobile engines to liquefied gas operation, etc.
The main objective is of course the conversion of automobile transportation
to electric power. Scientific research efforts should be concentrated
primarily in this direction. Our country, which is building interstellar
spacecraft, is certainly capable of constructing electric cars, that meet
all the requirements of economy, comfort, convenience of use, and hygienic
safety.
We shall not stop to discuss the problem of radioactive contamination of
atmospheric air, first because this is an entirely specialized problem, and
secondly because the road to the solution of the problem of protection of
atmospheric air from radioactive contaminants is a matter of international
policy, not medicine.
•v
However, it is indispensable to consider that aspect of the problem
which is connected with the development of industrial chemistry, a most acute
and topical problem of the modern era.
The role of chemistry in the acceleration of the building of Communist
society is very prominent. The 2 3rd Congress of the Communist Party of the
Soviet Union set forth high rates of development of the chemical and petro-
chemical industries. Chemistry and its products widely permeate all aspects
- 10 -
-------
of production and living conditions of the population. The production of raw
materials, semifinished products and finished articles from synthetic materials
will become on of the most common, complex, and largest sources of pollution of
the envirnoment and particularly atmospheric air, and will present the danger of
a harmful effect on man if hygienists, chemists, and engineers do not study
these compounds from the standpoint of their nature, biological characteristics,
and danger for man, and do not learn to determine them (which will require
specific and high sensitive methods of investigation) and to prevent their
presence in atmospheric air in concentrations adverse to man.
Interest in the pollution of the environment with chemical substances
has been increased throughout the world. In March 1963, the world Health Organ-
ization convened in Geneva a special international symposium on michrochemical
pollutants of the envirnoment. According to the data of this symposium,
hundreds of new organic compounds are being synthesized every year throughout
the world. Many of them find practical applications and constitute a source
of environmental pollution. The mere enumeration of the various areas of appli-
cation of new chemical compounds would require considerable space. They in-
clude various fertilizers, pesticides, defoliants, plastic articles used in
machine building, in housing, and in municipal services, new fabrics for cloth-
ing, shoes, objects for everyday use, packaging, effective washing agents,
detergents, bonding materials, synthetic drugs, vitamins, antioxidants for
fats, liquid fuel additives, etc.
The acuteness of the situation is made worse by the fact that the majority
of these organic compounds have not been studied from the toxicological and
physiological points of view, and no methods have been developed for their
determination in complex mixtures and low concentrations.
The possibility of their combined action, transformation in the environ-
ment, and the lack of their study from the standpoint of hygienic, carcinogenic,
and teratogenic effects compound the situation considerably.. Moreover, the
planners require data on the harmlessness of the new substances, which will
come in contact with the workers and engineers in the course of production,
with the population around the plants, and with millions of consumers utiliz-
ing the products of industrial chemistry in all parts of the country.
For this reason, chemistry should now be at the center of attention of
—public health science and in particular, atmospheric sanitation. The objectives
are large and, therefore, sometimes appear unreachable. However, this spurious
appearance should be resolutely rebuffed. It is necessary to mobilize all the
available resources, plan the sequence of the work closely, and allocate the
efforts appropriately.
It is perfectly obvious, for example, that additional resources should be
directed primarily at reinforcing scientific groups that have already formed
and proven themselves. Where qualified personnel, modern equipment and an
experimental base already exist, it is easier to achieve a marked increase in
the yield of practical research than in cases where all of this must be
created from scratch. It is therefore necessary to give support as rapidly
- 11 -
-------
as possible to scientific centers that are already doing work in the areas of
toxicology and chemistry, and in the area of new synthetic compounds and
pollutantB of the environment, including atmospheric air, associated with
their production.
Problems subject to investigation should be planned in a more serious and
active manner. Accelerated research methods should be adopted, with priority
given to those research areas that are absolutely indispensable for the formu-
lation of practical conclusions; studies that are less urgent and merely con-
firm the conclusions reached should be postponed.
The work of graduate students should be switched broadly to problems of
industrial chemistry, particularly synthetic chemistry. Priority and even
express handling should be accorded to the publication of materials dealing
with research in the area of industrial chemistry.
Atmospheric pollutants associated with the development of industrial chem-
istry have a number of characteristics which permit one to formulate the trend
and prospects of measures aimed at preventing their harmful action and the ways
of eliminating them.
For the most part, these are organic compounds characterized by a considera-
ble volatility, a distinct odor, and an irritant action on the mucous membranes,
particularly those of the upper respiratory tract. They can be neutralized in
the following ways.
1. Method of combustion in special furnaces. Unfortuantely, their combus-
tion sometimes requires the consumption of fuel because of the low concentration
of pollutants and an insufficient heat of combustion. A drawback of this method
is the destruction of valuable organic compounds instead of their recovery and
utilization in the national economy. However, in cases where no other means of
neutralization of these compounds exist, the combustion method remains available.
2. Compounds of this type usually condense readily, and hence, are capable
of being separated by means of units producing low temperatures. Once the
bulk of a compound has been separated by the condensation method, the remaining
amounts, which are no longer of any value, can be subjected to combustion.
3. Compounds of this type undergo purification by means of sorption fairly
readily, this being followed by desorption of the products obtained and regener-
ation of the filters. This is the chief method of recovery, and should be
widely employed in practice as one of the most universal means of controlling
the emissions of organic compounds.
The combination of these three methods makes it possible to find a satis-
factory solution to the most diverse emissions of synthetic chemistry.
In the case of substances marked by a low threhold of action, it is neces-
sary to find ways of eliminating them from the chemical process, with substitu-
tion of less toxic compounds or the introduction of a technology based on
fundamentally different methods not requiring the use of toxic substances.
- 12 -
-------
Thus, for example, the high-temperature heat carrier Dowtherm, widely
employed in synthetic chemistry and creating large zones of atmospheric pol-
lution around synthetic fiber plants, can be replaced by electric heating.
The development of this progressive method should be given considerably more
attention in the plans of the technological institutes concerned.
By comining the various enumerated trends in the prophylaxis of atmos-
pheric pollutants consisting of emissions of synthetic chemistry (altering the
chemical process, replacing noxious reactants by harmless ones, recovery,
combustion of residual effluent, etc.), one can always find the necessary
means of neutralizing industrial emissions. However, this requires two conditions:
1. Hygienists and toxicologists should become involved in the work of
scientific technological institutes dealing with the development of technolo-
gical schemes for new branches of industry.
2. Studies on the biological and toxicological effects of new chemical
compounds being placed in production, with the establishment of their maxi-
mum permissible concentrations and with the development of specific and highly
sensitive methods of their determination, should be intensified and acceler-
ated as much as possible.
As is evident from the above, the problem of atmospheric pollution has
become considerably more complex and difficult during its one-hundred-year
history. New problems have arisen which have gradually superseded the routine
considerations that used to prevail in this area during the first half of the
20th century.
Industrial chemistry and the struggle with automobile exhausts have be-
come the primary concern. In order to supply answers to the vital practical
questions, the priority of research in the area of industrial chemistry, must
be ensured.
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end of the
volume in the 1967 bibliography.
- 13 -
-------
• COMBINED EFFECT OF HYDROGEN FLUORIDE AND SULFUR DIOXIDE
ON THE BODY OF MAN AND ANIMALS
Z. Ya. Lindberg
Riga Medical Institute
From Akademiya Meditsinakikh Nauk SSSR. "Biologicheskoe deystvle i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 11, Izdatel'stvo "Meditsina" Moskva, p. 32-43, (1968).«¦
For quite some time, we have studied the pollution of the environment
by discharges of a large superphosphate plant producing superphosphate,
sulfuric acid (by the Mills-Packard process), and as a by-product, ammonium
fluoride.
The numerous studies have demonstrated that the superphosphate plant
constitutes a major source of pollution of atmospheric air with sulfur
dioxide, sulfuric acid aerosol, nitrogen oxides, and fluorine compounds
whose concentrations at a distance of 2000 m from the plant exceed the maxi-
mum permissible levels. The main ingredients of the discharges of the super-
phosphate plant are hydrogen fluoride and sulfur dioxide.
Our purpose was to study the combined action of low concentrations of
hydrogen fluoride and sulfur dioxide and to obtain data for substantiating
the maximum permissible concentrations of these substances when they are
jointly present in atmospheric air.
The isolated action of sulfur dioxide as well as hydrogen fluoride has
been adequately studied thus far, and the maximum permissible concentrations
of each of them in atmospheric air have been established and approved. In
order to substantiate the highest permissible single concentration of a com-
bination of these two substances, we determined the thresholds of olfactory
perception and reflex effect of sulfur dioxide and hydrogen fluoride on the
light sensitivity of the visual system.
F. I. Dubrovskaya established the olfactory threshold of sulfur dioxide
for the most sensitive persons at the level of 1.6-2 cig/rn^. According to
the data of M. S. Sadilova, this threshold for hydrogen fluoride is at a
level of 0.11-0.03 mg/m^. The determination of the threshold of smell of
the gaseous mixture was preceded by a separate verification of the thresholds
of olfactory perception of sulfur dioxide and hydrogen fluoride.
The observations were made on 17 persons. The threshold of smell of
sulfur dioxide for the most sensitive persons (eight people) was established
- 14 -
-------
at a level of 1.6 mg/m^; the concentration of 1.3 mg/nw was imperceptible.
The threshold of smell for hydrogen fluoride for the most sensitive persons
(six people) was established at a level of 0.04 mg/ra^, the maximum imper-
ceptible concentration being 0.02 mg/m^. Thus, the results of our studies
are close to the data of F. I. Dubrovskava and M. S. Sadilova. Data on
the determination of the odor threshold for mixtures of the concentrations
of sulfur dioxide and hydrogen fluoride studied are listed in Table 1.
Determination of the Odor Threshold of
Hydrogen Fl-jcride-Sulfur 3icxi.de Mixtures
Con centre ti or.
in ng/m'
Fractions of Threshold
During Isoleted Action
Sub of
fractions
of Thres-
hold for
Isolated
Action
Number of
Persons
Hydroger.
Fluoride
Sulfur
Dioxide
Hydrogen
Fluoride
Sulfur
Dioxide
1
J)OCC
0.5-i 1
•H
O C® C
UrMOJO
0.04
1,6
0,04 .
0,04 _ 1
1,6 ~ 1
2
17
—
0,03
'0,03 __
1,2 i0,04 1,0
0,2
— 0,70
1,45
.17.
—
0,02
0,
0,02
0.04 = 0,0
0.8 „
1,g -°,D
1
12
5
0,01
0,G5
0,01 „
0,04 °'2:i
0,G5
MT-=0-4
0,65
17
It is apparent from Table 1 that the minimum perceptible concentrations
in the mixture are 0.02 mg/m3 hydrogen fluoride and 0-8 mg/m3 sulfur dioxide
for a sum of relative concentrations equal to 1. The odor of the mixture is
not perceived if the sum of the relative concentrations is less than 1.
The results obtained suggest that a complete summation of the action cf
hydrogen fluoride and sulfur dioxide is noted in this case.
We studied the reflex effect of small hydrogen fluoride and sulfur
dioxide concentrations on the functional state of the central nervous system
by determining the light sensitivity of the eye under dark adaptation con-
ditions .
F. I. Dubrovskaya (1957) established that sulfur dioxide in a concen-
tration of 0.6 rag/m^ causes threshold changes (an increase) in the light
sensitivity of the eye. For hydrogen fluoride (according to the data of
M. S. Sadilova, 1965), the concentration having a threshold effect or. the
course of the dark adaptation curve is 0.03 rng/m^, while 0.02 mg/m3 is
inactive.
Our observations were made on four persons 26-38 years old, most
sensitive to the threshold of smell, with a normal visual acuity.
- 15 -
-------
A procedure recommended by V. A. Ryazanov, K. A. Bushtuyeva arid Yu. V. Novikov
(1957) was employed.
To measure the light sensitivity of the eye, we used an "ADM" adaptometer.
The hydrogen fluoride-sulfur dioxide mixture was supplied in the 15th minute
of dark adaptation for 5 minutes. The light sensitivity of the eye was
determined every 5 minutes up to the 30th minute and in the 40th minute. The
tests were made three times with each concentration.
We carried out observations with the following concentrations: sulfur
dioxide 0.3 mg/m^, hydrogen fluoride 0.015 mg/m3> sulfur dioxide + hydrogen
fluoride in concentrations of 0.25 + 0.01 mg/m3, and sulfur dioxide + hydro-
gen fluoride in concentrations of 0.3 + 0.015 mg/m3.
In all four subjects, the concentrations of sulfur dioxide (0.3 rag/m^)
and hydrogen fluoride (0.015 mg/m^) and also the sum of sulfur dioxide and
hydrogen fluoride (0.25 + 0.01 mg/m^) did not cause any changes in the light
sensitivity of the eye.
During inhalation of sulfur dioxide with hydrogen fluoride in concen-
trations of 0.3 + 0.015 mg/m^, the light sensitivity of the eye in the 20th
and 25th minutes increased considerably when compared with the normal sensi-
tivity in three subjects (Fig. 1) and in the 20th minute in one subject.
U5000
U0000
35000
I 30000
• H
§
$ 25000
§ 20000
c .
>> J 5 000
•w*
>
'0000
c
Q)
V,
£ 5000
te
0
5' to' >5' to' es" JO' 35' «0'
Time, ninutes
Fig. 1. Change in the light sensitivity of the eye in
subject L.
Pure air
HP-0.0!5 ng/n3
—• — •— <50?"0.3 iqg/r.^
— HF-0.0t v%lr?>'SQ}-0,?5 mg/m^
— — —HF-O.OlS^l^'SOz -03
- 16 -
-------
Statistical treatment of the data obtained showed that the apparent
changes of the light sensitivity of the eye were significant. Thus, the
threshold of the reflex effect of the sulfur dioxide - hydrogen fluoride
mixture on the light sensitivity of the eye was 0.3 + 0.015 mg/tn3. The
fractions obtained by dividing the concentrations studied into threshold
concentrations during isolated action add up to 0.74 for the first mixture
and to 1 for the second mixture. The only active mixture was the second
one, whose inhalation caused first an increase and then a decrease in the
light sensitivity of the eye. A mixture of sulfur dioxide and hydrogen
fluoride of lower concentrations with a sura of the parts of threshold con-
centrations equal to 0.74 was found to be inactive for all the subjects.
In this case, there is a total summation of the effects of each substance.
In order to detect the resorptive effect of low concentrations of
hydrogen fluoride and sulfur dioxide in a mixture, we subjected 90 white male
rats weighing from 80 to 95 g to chronic exposure for three months. The
animals were divided into six groups.
The concentrations of the substances in the chambers during the period
of exposure are shown in Table 2.
Table 2
Concentrations of Sulfur Dioxide ar.d Hydrogen Fluorid-.
in ExpericentaJ Chanbers
N'uaber
.of
Specified Concen-
trations in bb/b?
Actual Concentrations in mg/ra'
Chamber
Sulfur
Dioxide
Hydrogen
Fluoride
Sulfur Dioxide
Hydrogen Fluoride
1
II
Ml
IV
V
5,0-
0,15
0,25
5,0
. 0,3
0,01
0,01
0,3
5,01 ±0,063
0,147 + 0,0004
0,249 + 0,003
4,99 ±0,043
0,297 + 0,0708
0,0'99 + 0,00039
0,0099 + 0,00038
0,3 + 0,0048
VI
Control
For one month prior to the exposure, the general state of health of the
animals was observed, and the following indices were determined: their
weight, the motor chronaxy of antagonist muscles, cholinesterase activity,
amount of coproporphyrin in the urine, morphological composition of peripheral
blood, and the amount of calcium, inorganic phosphorus, sugar, and catalase
in the blood. These indices were also studied during the exposure of the
animals. Rats of the third, fourth and control groups were healthy, active,
and gained weight normally in the course of the experiment.
Rats of the first, second and particularly fifth group were less active.
Beginning with the fourth week of exposure, the fur of the rats in the second
- 17 -
-------
and fifth groups lost its natural luster and bristled up. Irritation of
the mucosa of the eyes was observed in rats of the fifth group.
The motor chronaxy of the flexors and extensors of the shin was deter-
. mined by means of an 1SE-01 pulsed electronic stimulator. The tests were
carried out on five rats of each group once every 10 days under the same
conditions.
Fig- 2 shows the chronaxy of antagonist muscles of rats exposed to
hydrogen fluoride, sulfur dioxide and their mixture (average data for the
groups).
Reliable changes in the ratio of chronaxies
of extensors and flexors were noted in rats of
the second and fifth groups in the fourth week
of exposure, and in rats of the first group,
in the fifth week. In rats of the third and
fourth groups, a change in the ratio of chron-
axies of antagonist muscles was observed in
the third month of exposure.
Cholinesterase plays an important part
in the process of functional activity of the
nervous system. The cholinesterase activity
in the blood was studied on a photoelectro-
colorimeter in five rats of each group once
every ten days by using the Fleischer-Pope
method as modified by N. N. Pushkina and
N. V. Klimkina. The blood for the study
was taken from the tail vein by incising
the tail, asepsis rules being observed.
The cholinesterase activity was expressed
in micromoles of acetylcholine per 2 ml of
blood.
The change of cholinesterase activity
in rats of different groups is shown in
Fig. 3.
123U561Q910 lit? 13
Extensors
Flexors
Fig. 2. Motor chronaxy of antagonist
muscles in rats of different groups.
In animals of the first, second and fifth
groups, beginning with the third week from the
start of exposure, a statistically reliable
increase of cholinesterase activity was observed, and at the end of the
experiment, there was a decrease of this activity.
- 18 -
-------
<« 0.8
&
CO
U
-------
pneumonia (Fig. 6)
.5
1-4
o.x>
u
O Vj
a o
o
u »
o o+>
o Ojz
rMtf)
Cm >H
a*
1."
'¦2
'0
. as
»
05
,/ v Z\
A, T
v / \ / / —^
/ ! V /
0
2 3 U 5 6
_ ¦ Time,, weeks
rirst series '
Second series -»-»*¦ • Fourth series
Third Series Fifth series
9 >0 ti V
.Sixth (control)
series
rig. 4. Content of coproporphyria in the urine of animals of all
groups.
Rats of the fifth group had an inflammatory infiltration in the
stroma of the kidneys, primarily around the glomeruli (Fig. 7). Histo-
chemical analysis of the respiratory organs and kidneys in rats of the
first, second, and, particularly, fifth groups showed the accumulation
of neutral mucopolysaccharides in the walls of the bronchi and in the
glomeruli of the kidneys.
J*-?
*4t S « , • V V, T ^
¦ v< :
t* i>'
ik
Fig, 5. Hats of fifth series. Changesin the lungs. Magnifi-
cation ^0 x 10. Inflammatory infiltration,in the stroma,
rteralveolar septa (interstitial pneunonia), sosae
thickening of mte
tises emphysema and edema.
Peribronchitis
- 20 -
-------
'iT aL i»' vp,'v-
'5* •-4;vV>'sr1?: ^
. y/*-* ^ £(&z&~ "\i
r -f* i. •*
«4ss
,_ W Sw*
. fj&k iM $«¦>*
•* *W?-VAf ' -:..'
¦ft. . j-^c? ¦ •• ¦ ¦¦ :¦>
- • "** v* *
BF ^ ^ T • . i.-^ '*..•,
I'r- > * •. «- v' w- •- / ,7: .y.4
<""^1
,,»
J-ot'
Fig. 6. Rats_ of fourth series. Changes in lur.gs. Occasional
emphysema, thickening of interalveolar septe »ith cellular infil-
tration (as reaction to irritation).
»Js* ••••'• ;f; \ • «
jhvN'I *" v% ^J*5%'•• . V -"• *- ;•-'
€r^ ** * '«»..~ •-•*"- •' * - /• : - • •: ,*5
• i.-'» • .? * !»t .'- 4- *';*# -«-'•• r- -*.•,-*? ¦ «< " > -¦; . ,• • . *
,V iv'-*-' ^
*^*ssnA»*sS* ^-•-;v.r;
~1
¦I
• -1
4
— ,r-» * ~
%.%¦ fS^i^' .;^:'.': •>*<'-V
->Xik,A'Jrlhi i,!rA
Fig, 7. Rats of fifth series. Changes in kidneys. Magnification
4C x 10. Photograph shows inflaianatory infiltrate in strosa,
chiefly around glomruli.
- 21 -
-------
fi->^ ...%• .•
6 . .--v .
• '"k,••. » *'
i
,.J
Fig. S. Strietion cf enamel of lower inoisors ir. experimental
rats of fifth series.
An increase of fluorine was found in the bones of rats of the second
and fifth groups. The amount of fluorine in the bones of rats of the
second group was 6.6 mg, in rats of the fifth group 6.8 mg, and in the con-
trol, A.8 mg per 100 g of dry substance. In rats of the second and fifth
groups, early signs of fluorosis were detected in the form of striation of
tooth enamel (Fig. 8).
Thus, a prolonged inhalation of a mixture of 0.3 mg/m^ hydrogen fluoride
and 5 mg/m3 sulfur dioxide caused considerable changes in the rats: in the
ratio of motor chronaxies of antagonist muscles; in cholinesterase activity;
in the excretion of coproporphyrin with the urine; an increase in the number
of leucocytes; a decrease in the number of erythrocytes and amount of blood
hemoglobin; and also changes in the lung tissue and in the kidneys.
In rats that inhaled a mixture of 0.01 mg/m^ hydrogen fluoride and
0.25 mg/m3 sulfur dioxide (inactive concentration of the mixture for short-
term inhalation) and also in rats exposed to a mixture of hydrogen fluoride
and sulfur dioxide at the level of maximum permissible concentrations
(0.01 mg/m3 for hydrogen fluoride and 0.15 mg/m^ for sulfur dioxide), the
changes in cholinesterase activity, in the excretion of coproporphyrin with
the urine and in the morphological composition of the blood were insignificant.
Changes in the ratio of chronaxia of the antagonist muscles were reliable.
A histological examination of the organs of the animals of these series showed
significant changes in the lungs in the form of a thickening of the alveolar
septa, areas of interstitial pneumonia, and emphysema.
Thus, the studies showed that the combined presence of sulfur dioxide
and hydrogen fluoride in the atmospheric air of populated areas at the level
of the existing mean daily maximum permissible concentrations for each sub-
stance is inadmissible.
-------
The total mean daily concentration of sulfur dioxide and hydrogen
fluoride present together, expressed in fractions of the maximum per-
missible values established for each of them, should not exceed unity.
Conclusions
1. The effect of complete suamation was established in a study of
the reflex and resorptive effect of a mixture of sulfur dioxide and hydro-
gen fluoride.
2. When sulfur dioxide and hydrogen fluoride are present together
in atmospheric air, their highest single and mean daily maximum permissible
concentrations, expressed in fractions of the individual maximum permissible
concentrations, must not exceed unity.
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end of the
volume in the 1968 bibliography.
- 23 -
-------
NEW DATA FOR THE VALIDATION OF THE MEAN DAILY MAXIMUM PERMISSIBLE
CONCENTRATION OF HYDROGEN FLUORIDE IN ATMOSPHERIC AIR
M. S. Sadilova, E. G. Plotko, and L. N. Yel'nichnvkh
Sverdlovsk Institute of Labor Hygiene and Occupational Diseases
* Prom Akademiya Meditsinakikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 11, Izdatel'stvo "Meditsina" Moskva, p. 5-15, (1968).
In the available literature on the toxicological characteristics oi
inorganic fluorine compounds, there are no convincing data on changes
induced in the organism by the inhalation of comparatively low fluorine con-
centrations, and no data whatsoever on the threshold and inactive concentra-
tions. The permissible fluorine concentrations of atmospheric air adopted
earlier were not subjected to experimental verification and no account was
taken of possible differences in the biological action of the various
fluorides (S. V. Miller, 1955). And yet, the national economy plan specifies
a further development of aluminum plants, the production of superphosphates,
the expansion of the production of fluorine salts and of concerns for the
enrichment of fluorspar and other branches of industry utilizing fluorine
compounds. In view of these objectives, the problem of a scientific valida-
tion of the safe level of fluorine content in the air of populated areas
assumes a major importance.
The present report and two other studies of the present collection
present the results of several series of investigations which provide a
toxicological assessment and make it possible to determine the parameters
of the noxious effect on the organism of gaseous and pulverized fluorine
compounds marked by different solubilities in biological media.
In 1965, on the basis of experimental studies, we proposed a permiss-
ible mean daily concentration of hydrogen fluoride (HF) in atmospheric air
at a level of 0.01 mg/m^, which was shown by tests to be the maximum inactive
concentration.
In order to refine certain problems of the mechanism of action of
fluoride ions and to study the phosphorus-potassium metabolism by using
isotopic tracing, we continued the studies with HF at the level of the con-
centrations investigated earlier: 0.10-0.03 and 0.01 mg/m^. In the course
of exposure of the animals, the following quantities were also refined:
1) activity of alkaline phosphatase of the blood; 2) degree of excretion of
fluorine from the body through the kidneys and the gastrointestinal tract;
3) accumulation of fluorine in the teeth and bone tissue.
- 24 -
-------
All the conditions of the previous (^irst) experimental series were
observed: 1) the HF concentrations in the chambers during the period of
exposure were 0.10 it 0.002, 0.03 i 0.001 and 0.01 — 0.0009 mg/m^; 2) the
experiment involved female rats two months old kept on the same food and
water diet as the animals of the first series; 3) the animals were sub-
jected to a five-month round-the-clock exposure followed by a one-month
period of recovery.
In the first series of studies, inhibition phenomena in the central
nervous system were observed in animals of the first and second groups.
In order to determine better the mechanism of the inhibitory effect of
fluorine ions on the central nervous system, we studied the activity of
brain and blood cholinesterase, the content of pyruvic acid in the blood,
the content of sulfhydryl groups in the brain, and the state of nerve cells
and interneuronal connections.
It is well known that cholinesterase, which decomposes acetylcholine
into choline and acetic acid, plays an important part in the synaptic
transfer of nerve impulses. The activity of pure cholinesterase present
in the gray matter of the brain was studied at the end of the exposure and
at the end of the one-month recovery period (the activity of the enzyme was
determined in 7-8 animals of each group after a Fleischer-Pope homogeniza-
tion as modified by N.. N. Pushkina and N. V. Klimkina). Our studies did
not show any differences in the activity of the brain enzyme in the
"fluorinated" and control animals. However, we did record a statistically
reliable depression of the blood cholinesterase in animals of the first
and second groups in the first two months of exposure, at its end, and
after the recovery period.
In the normal process of synthesis of acetylcholinelike substances
in cells, conjugated synthetic processes take place between choline, which
is a product of phosphatide metabolism, and a product of carbohydrate
metabolism capable of acetylating choline. The formation of acetylating
agents in the conversion cycle of carbohydrates proceeds via the stage of
pyruvic acid (Kh. S. Koshtoyants). Our determination of pyruvic acid by
the Friedemann-Haugen method confirms the presence of a "fluoride block"
in animals of the first and second groups. An increased content of pyruvic
acid was also observed one month after the end of exposure (Table 1). These
data attest to the depression of carboxylase, an enzyme decomposing pyruvic
acid.
Many researchers have shown that an enormous number of enzymes partici-
pating in the conversion cycle of pyruvic acid and also cholinesterase
require for their activity the presence of sulfhydryl groups in the protein
component of their molecules. The binding or blockage of sulfhydryl groups
depresses the activity of many enzymes of protein and carbohydrate metabo-
lism. According to Koshtoyants, they are of major importance in normal
- 25 -
-------
processes of stimulation and inhibition of the nervous system, since the
activity of the receptor is determined by the presence of free sulfhydryl
groups in the protein molecule. In the determination of the content of
sulfhydryl groups by the Kolthoff and Harris method, we observed their
decrease in the brain tissue at the end of exposure in animals that inhaled
HF in a concentration of 0.10 mg/m^. After the one-month recovery period,
the content of sulfhydryl groups returned to normal.
After the completion of exposure of animals in the first group, a study
of the histological specimens of the cerebral cortex showed a distortion .of
the nerve cells, disappearance of the Nissl substance, and pyknosis and
lysis of the nuclei. Extended areas with damage to the apical dendrites
were observed. The dendrites had beaded enlargements, and were broken down
into fragments: the specimens showed black dots that were not connected to
each other by strands of cytoplasm. Histopathologic changes of the nerve
cells and dendrites in animals of the first group were also observed after
the one-month recovery period.
There were also changes in the cerebral cortex in animals of the
second group. However, these changes were less pronounced. Nerve cells
with granular cytoplasm containing clear vacuoles were found only occasionally.
Cells were observed whose dendrites had enlarged beads. Changes of neurons
in the presence of 0.03 mg/m^ HF were reversible in character.
In animals inhaling HF in a concentration of 0.01 mg/m^, no pathologic
changes were detected in the neurons.
Table 1
Concentration of Pyruvic Acid (in milligraa-percer.t) in the
Blood of the Animals
Group of
Animals
At the End of
Chronic Exposure
At End of Or.e-Month
Recovery Feriod
Statistical Criteria
n
X±Sx
. t
P .
n
X±Sx j l
P
First
Second
Third
Control
6
6
6
6
4,8±0,05
4,2 + 0, 1c
3,6 + 0,2f
3.5 + 0.14
8,74
3,41
a,32
<0,001
<0.01
>0,05
6
6
6
6
4,6^0,14J5,85
4,2 + 0,1613,6:
3,7±0,1G 1 ,
3,4±0,15j —
<0,001
<0,01
>0,05
Thus, the shifts in the functional state of the central nervous system
which we observed in the first series of experiments resulted from the
inhibitory influence of fluoride ions on enzyme systems involved in the trans-
fer of nerve stimulation and from the histopathologic damage to nerve cells
and interneuronal connections.
- 26 -
-------
Disturbances in the phosphorus-calciur metabolism and in the architectonics
of bones are attributed by researchers to the inhibition of phosphatase activity.
From esters of phosphoric acid phosphatase liberates inorganic phosphorus, which
combines with calcium to form the phosphorus-calcium salts necessary for build-
ing bone tissue. Kutscher and Wust, Hassart and Dufait, Smith, and others ex-
plain the mechanism of phosphatase depression by fluorine by the fact that
magnesium, which enters into the enzyme complex and is the enzyme activator,
forms a nondissociable form on combining with fluorine. The degree of phos-
phatase inhibition does not always correspond to the concentration of fluorine
ions; high concentrations either do or do not cause a lesser inhibition effect
than intermediate or relatively low concentrations. On the basis of their
studies, Reiner, Tsuboi, and Hudson believe that fluorine, by undergoing poly-
merization in the organism, forms a dimer and a tetramer. The dimer is formed
when low concentrations of fluorine enter the organism, and a tetramer is
formed in the presence of high concentrations. The dimer combines with the
active center of the enzyme. The tetramer also combines with the active
center, but may be replaced by a substrate, and the enzyme inhibition effect
decreases at high fluorine concentrations. We studied the activity of alka-
line blood phosphatase under dynamic conditions according to G. K. Shlygin
and S. Ya. Mikhlin in six animals of each group. In the presence of hydrogen
fluoride in a concentration of 0.10 mg/m^, the inhibition of alkaline phos-
phatase of the blood was observed only during the second month of exposure of
the animals. A more pronounced effect was detected as a result of inhalation
of HF in a concentration of 0.03 mg/m^: inhibition of the enzyme was observed
during the second, third, and fifth months of exposure. Even as low an HF
concentration as 0.01 mg/m3 can depress the enzyme activity: a statistically
reliable shift was observed at the end of the fifth month of exposure.
We studied the rate of phosphorus and calcium metabolism in the body by
means of isotopic tracers. The radioisotopes p32 and Ca^S in tracer doses
were introduced into the animals intraperitoneally at the end of chronic
exposure and at the end of the one-month recovery period. Twenty-four hours
after the introduction of the isotopes, the animals were sacrificed by decapi-
tation. The analysis was performed on the teeth, humerus, femur, blood,
and liver. The teeth and tubular bones were thoroughly cleaned to remove the
soft tissues. All the specimens were then weighed, brought to a constant
weight at 105°C., and ground into a powder. From each specimen, portions
weighing 30 mg were evenly spread over aluminum targets. The activity was
measured with a type B counter. The total activity produced by the radiation
of p32 and Ca^5 was determined first, then the activity of Ca^5 was completely
cut off with an aluminum filter, and the activity due to P32 was calculated.
In calculating the activity, necessary corrections were introduced for the
natural decay of the isotopes, absorption in the specimens, etc. The isotopic
tracer method revealed a statistically reliable retardation of the inclusion
of phosphorus and calcium in the biological substrates studied. More pro-
nounced changes were found in the phosphorus metabolism in animals of the
first and second groups; phosphorus was included more slowly in all the speci-
mens studied than in the control. Even when the lowest EF concentration
- 27 -
-------
Inclusion of Radioisotopes P52_and Ca^5 in Tissues and Blood of Animals at find of Kiw-Vonth'-:xr^:::v
to HF (in 1 g of Moist Tissue in Percent, of Activity introducr-d into Body)
Group of Animals
Biosubstrate
Isotope
•Statistical
Criteria
first
1
Second |
Third ' j
Control
Teeth
pai
.
X±Sx
I
7
0,75+0,12
4,24
<0,001
6
1,00+0,05
2,56
<0,05
8
1,20 + 0,10
1,00
>0,05
,s
1,35 r. 0.12
Ca<»
n
X±Sx
t
P
8
3,00±0,36
1.87
>0,05
7
3,34 + 0,25
1,35
>0,05
7
3,88+0,26
0,01
>0,05
6
3,82-0,25
Femur
psi
n
x±s*
t
P
7
0,43±0,05
6,66
<0,001
7
0,52 + 0,11
3,18
0,01
8
0,72x0,02
3,35
<0,01
8
0.93 ± 0,06
Ca4S
n
X±Sx
t
P
8
1,66 + 0,11
5,18
<0,001
7
1,88+0,22
2,54
<0,05
7
2,54i0,11
0,74
>0,05
S
2.SS-0.16
Humerus
p 32
n
X±Sx
t
P
7
0,45 + 0,05
4.08
<0,002
7
0,56 + 0,09
2,36
<0,05
8
0,69 + 0,04
1,83
>0,05
8
0,85±0,08
Ca<»
n
X±Sx
t
P
8
1,96±0.14
3,44
<0,01
6
2.1.7+0,28
2,00
>0,05
8
2,59 + 0.13
1,19
>0,05
8
2,91 ±0,24
z
31cod
pjj
n
X±Sx
t
p
7
0,060±0,006
7,00
<0,001
6
0,060 + 0,010
5,00
<0,001
6
0,090 + 0,005
2,86
<0,05
8
o,:;o+o,oo5
•
Ca«»
X±Sx
t
P
8
0,030+0.001
1,40
>0,05
6
0,030+0,004
1,20
>0,05
6
0,040 + 0,007
0
>0.05
8
0,040-0,007
Liver
P32
.
j X + Sx
1 t
'
7
0,61+0,05
4,07
<0,002
6
0,70+0,04
3,54
<0,01
• 8
0,78 + 0,05
2,30
; <0,05
i
s
I 0,96 + 0,06
!
- 28 -
-------
(0.01 mg/m3) was acting on the organism, a delayed inclusion of radio-
active phosphorus was noted in the femur, liver and blood. Disturbances
in the calcium metabolism were noted only in animals of the first and
second groups (Table 2). Changes in the phosphorus-calcium metabolism
continued to be observed in all three "fluorinated" groups of animals and
after the one-month recovery period. It is necessary to postulate that the
observed mottling of tooth enamel and the histologic changes in the bone
tissue in animals of the first and second groups resulted from a disturbance
of the phosphorus-calcium metabolism.
Repeat determinations of the fluorine content of the urine in the
second series of experiments confirmed a high level of excretion of fluorine
from the body. A higher fluorine concentration, which occurred during the
second month of exposure in the urine of certain groups of animals, was re-
tained until the end of the inhalation period. The fluorine content of the
urine was almost directly proportional to the concentration of hydrogen
fluoride in the air inhaled. A reliable increase of fluorine in the urine
and in animals of the third group was observed.
We were interested in the problem of the level of fluorine excretion
from the body through the kidneys and through the intestines. These studies
were made at the end of the chronic exposure on animals of the first group
and on the controls. It was found that in animals subjected to the inhala-
tion of hydrogen fluoride, over 78% of the fluorine is excreted through the
kidneys, and 15% through the intestines. In the control group, the reverse
relationship was observed. These data lead to the conclusion that the
gaseous fluorine that has entered the body is excreted through the kidneys
as a result of a complete absorption in the respiratory organs (I. D. Gadaskina)
and penetration of the general circulatory system. The fluorine retained in
the body is deposited in the teeth and bone tissue. A higher degree of fluor-
ine accumulation was observed after exposure to the higher hydrogen fluoride
concentration (Table 3).
After the chronic exposure was completed, histological analyses were
darried out which showed that HF in 0.10 mg/m^ concentration causes changes
in all the internal organs.
In the study of the upper respiratory tract, the main changes were
observed in the mucous membrane and submucous layer. The mucous membrane
is thinned out in some places and swollen in others. In some areas the
epithelium is absent, cast off, or lies in the lumen of the air tube.
Leucocytes, erythrocytes and lymphocytes are visible among cells of the cast-
off epithelium. Large leucocytic clumps are present on the mucous membrane
that remains. In the submucous layer, the capillaries and vessels are greatly
enlarged. There are hemorrhages. The same type of changes were observed in
the bronchial epithelium. In some areas, the cast-off epithelium fills the
bronchial lumen, obstructing the bronchi completely. In the submucosa of the
- 29 -
-------
Table 3
Fluorine Content of Teeth, Humerus and Femur (in Milligram-Percent of Moist Substance) after Exposure
of the Animals to Hydrogen Fluoride.
Teeth
Humerus
Femur
Animal
i
Statistical Indices
n
X+Sx
t
p
n
X±Sx
t
P
n
X±S*
t
p
First
8
75,5 + 2,19
23.66
<0,001
8
73,1 ±1,67
27,76
<0,001
8
80,3+2,76
25,5
<0,001
Second
7
33,0+1,47
8,96
<0,001
7
37,9±1,96
10,31
<0,001
7
23,9+3,49
3,33
<0,01
Third
8
17,3+1,38
0,91
>0.05
8
18,5 ±2,29
1,96
7>0,05
8
13,2±2,12
1
>0,05
Control
8
15,6+1,27
"""
8
13,3 ± 1,36
—
8
12.3±0,61
—
-------
bronchioles, a polymorphous cellular infiltration is visible which sometimes
destroys the epithelium and comes out into the bronchial lumen. An acute
hyperplasia of the peribronchial lymphatic nodes resulting in the deforma-
tion of the bronchi was detected. The hyperplastic nodes extend under the
mucous membrane, causing it to bulge out in the lumen and form outgrowths,
thus destroying the muscle layer of the bronchi, causing its segmentation and
atrophy. This in turn causes the formation of bronchiectases. The inter-
alveolar septa are enlarged because of the dilatation of the capillaries and
their congestion with erythrocytes, and also the appearance of lymphocytes
and histiocytes in them. The space of many alveoli is filled with a transudate
containing cast-off epithelial cells, leucocytes, and erythrocytes. In some
areas, there are so many erythrocytes in the alveolar lumen that the contents
assume a hemorrhagic character. A perivascular edema is observed. In isolated
cases, bronchopneumonia is superimposed on the process described.
In the liver, kidneys, adrenal glands, and spleen, congestion phenomena
are observed in the form of an expansion and overfilling of capillaries with
erythrocytes and the formation of hemorrhages. Degeneratively dystrophic
changes are represented by pyknosis of the nuclei of renal convoluted tubules
and of liver and Kupffer cells. In addition, a development of infiltrative-
proliferative processes around the liver vessels is noted. The formation of
clear foci consisting of erythrocytes, cellular fragments with pyknotic nuclei,
and reticuloendothelial cells was observed in the liver.
Congestion in the red pulp, a certain loss of follicles and an increase
of the reticuloendothelial elements in then were observed in the spleen.
Changes were also noted in the cardiac muscle, i.e., the disappearance
of transverse striation and disjunction of the myofibrils.
A hydrogen fluoride concentration of 0.03 mg/m^ causes changes in the
liver only. The nature of the injury is similar to the above. No histopath-
ologic changes in the internal organs were detected following exposure to a
hydrogen fluoride concentration of 0.01 mg/m^.
Conclusions
1. Round-the-clock exposure to hydrogen fluoride concentrations of
0.10 and 0.03 mg/m3 causes inhibition in the central nervous system, decreases
the activity of a number of enzymes, impairs the phosphorus-calcium metabolism,
and causes the accumulation of fluorine in the body and damage to the internal
organs and bone tissue.
2. A hydrogen fluoride concentration of 0.01 mg/m^ should be regarded as
the threshold concentration. Its effects on the body of the animals showed
changes in the phosphorus metabolism only (inhibition of alkaline blood
- 31 -
-------
phosphatase and a delayed inclusion of radiophosphorus in bone tissue, liver,
and blood at the end of a five-month exposure of the animals).
3- The mean daily maximum permissible concentration of hydrogen fluoride
in the air of populated areas, adopted earlier as 0.01 mg/m^, should be lowered
to 0.005 mg/m3.
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end of the
volume in the 1968 bibliography.
- 32 -
-------
•SANITARY EVALUATION OF FLUORIDES READILY SOLUBLE
IN BIOLOGICAL MEDIA
M. S. Sadilova and E. G. Plotko
Sverdlovsk institute of Labor Hygiene and Occupational Diseases
"From Akademiya Meditsinakikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova
Vypusk 11, Izdatel'stvo "Meditsina" Moskva, p. 16-26, (1968).
Having studied the nature of the action of hydrogen fluoride, we con-
sidered it necessary to investigate fluoride salts as well, since we sus-
pected that gaseous and powdered fluorine compounds may differ in the
degree of their toxic effect on the body. These differences may be due to
different levels of retention of fluorides in the respiratory organs and
th£ir different solubilities in biological media.
It is known from studies made by I. D. Gadaskina and T. A. Shtessel'
that HF is completely absorbed in the respiratory organs. The behavior of
fluorine salts present in a finely dispersed state in air is partially ind
cated in Table 1, taken from a handbook on the toxicology of radioisotopes
Table 1
Distribution of Fine Particles in the Respiratory Org&ns of a
"Standard" Person.
Compounds
Nature of
Distribution
Headily Sol-
uble Com- Other Co>-
pounds, $> pounds, %
Excreted with exhaled air
Deposits in upper respiratory trest
- then ingested
Deposits in the lungs
25
25
50
251
50
2aJ
1 This part assimilated in the tody
2 Of this, one-half is excreted by the lungs ar.d ingested in
the first 24 hours, sc that the fraction of swallowed particles
increases to 62.9$. The remaining 12.5# reach the tody fluids.
It is evident from Table 1 that for one and the same concentration
of gaseous and powdered fluorine compounds in air, different amounts of
F~ will enter the human body.
-------
Table 2
Solubility of Fluorides in Different Media (Average Data for
Three Observations)*
riae .of
Withdrawal of
Samples for
Analysis
3inger-Tyrode
sol'jtion (pK = 7.2)
G
astric Juice
CpH = 2.5)
NaF
cT
<
| Na.AlF,
tT
5
u.
e)
Z
e?
<
U.
<
Z
M
Ci.
u.
Percent of Amount of Fluorine Introduced
After 1
hour
94.9
3.9
0,0
0.0
48,4
0,0
0,0
0,0
> 2
hours
98,8
3,9
2.9
0,0
48,4
0,0
0,0
•0,0
> 5
hours
98.8
3.9
2,9
0,0
48,4
0,0
0,0
0,0
» 24
hours
96,6
5,9
3,9
0,0
82.5
0,0
0,0
0,0
» 5
days
100,0
5,9
12.9
1,9
82,5
9.7
4,8
4,8
» 10
S
100.0
21,2
29,9
3,9
100,0
9.7
4,9
21,1
» 20
1
100,0
38,7
37,5
21,2
100,0
9,9
9,7
35,0
^ Tht' minimum a-otm^s were taken for the study of solubility -
0.16 mg of F in 100 nl of solution.
The data we obtained on the degree of solubility of different fluorides
in biological media are listed in Table 2.
The highest solubility in biological media was displayed by sodium
fluoride. It is completely soluble in weakly alkaline media. In acid
medium (gastric juice) during the first five hours, which are of practical
importance, the solubility of NaF was also higher - 48.4% of the fluorine
introduced. As the period of exposure increased, the solubility of NaF in
the gastric juice increased to 82.5-100%.
Aluminum fluoride and cryolite are very similar in their degree of solu-
bility in the Ringer-Tyrode solution. During the first 24 hours, the solubil-
ity of aluminum fluoride and cryolite is equal to 4-6% of the amount of
fluorine introduced; calcium fluoride is insoluble. As the period of contact
increases, the solubility of aluminum fluoride and cryolite increases, and
on the 20th day reaches 38%; the solubility of calcium fluoride is slightly
lower. Aluminum fluoride, cryolite and calcium fluoride are insoluble in
gastric juice during the first 24 hours. These data permit the assumption
that the NaF retained in the body dissolves completely in certain biological
fluids. Aluminum fluoride, cryolite, and particularly calcium fluoride will
not dissolve completely in biological fluids and hence may have a lesser
toxic effect than HF or NaF.
To study the fluorine salts, we set up a special experimental apparatus
which enabled us to obtain a condensation aerosol. The experimental influence
of fluorine condensation aerosols on the body frequently corresponds to actual
conditions (aluminum and other industries where high temperature processes
- 34 -
-------
are employed). The experimental assembly was equipped with automatic
blocking of the air supply instruments and with signaling that came on when
the operating conditions of its main units were impaired.
Table 5
Lignt Sensitivity of ^he Eye During Inhalation of Sodium Fluoride
_ J>J! Sut.'ect L.
h
S -
S
o
O"-* s
SR -P
Statistical
Criteria
Observation lime ir. Minutes^
20
25
30
40
Pure Air
X + S.v
126,3 + 7,6
¦ 183,0 ±21 ,c
223,5i 17,9
281,9 + 31,5
10,07
X±Sx
t
P
198.1+8,3
6,35
0.01
195.4±1'1,!
0,51
>0,05
324,4 + 48,0
1,96
>0,05
399,4±18,8
3,2
<0,05
0.05
X±Sx
t
P
154,1+1,1
3,61
0,02
166,1-2,0
0,78
>0,05
215,3± 16,5
0,34
>0,05
299,3 + 40,3
0,34
>0,05
0,03
X + Sa
t
P
134,0 + 3,9
0.S9
>0,05
210,1 +9,2
1,41
>0,05
259,7+ 10,&
1,73
>0,05
304,5+ 17,3
0,63
>0,05
percent of 15-r.inute dark adaptation, taken as ICO.
In order to standardize the highest single maximum permissible concen-
tration of NaF, the reflex effect of the latter on the light sensitivity of
the eye was studied by using the common method of dark adaptation in three
persons with normal vision. The study of the reflex effect of NaF showed
that a concentration of 0.07 mg/m^ causes an increase of the light sensi-
tivity of the eye in all the subjects, and a concentration of 0.05 rag/m3
does so in one person only. A concentration of 0.03 mg/ir.3 of NaF did not
cause any deviation of dark adaptation (Table 3).
To compare the results of the study with those obtained in the experi-
ments with HF, a chronic exposure of the animals to sodium fluoride was also
carried out round the clock over the course of five months. The NaF concen-
trations were checked four times a day. The withdrawl of air samples from
the chambers for the purpose of analysis for the degree of absorption of F~
was carried out by using dinitrocellulose membrane filters No. 2 in combin-
ation with absorbers filled with double-distilled water. Fluorine was
determined by the alizarin-thorium procedure (sensitivity of the method,
0.001 mg/m^). Since in the chronic exposure of animals the HF concentra-
tion of 0.01 mg/m^ was found to be the threshold value, only two concentra-
tions, 0.10 and 0.03 mg/m^, were studied in the experiment with NaF, which
- 35 -
-------
is incompletely retained in the body and in our view should be less toxic.
The actual NaF concentrations in the chambers during the period of exposure
are indicated in Table 4.
lable U
Sean Daily NaF Concentrations (in terns of F~)
During the Period of Exposure.
Chamber No. 1; Given Concentration
0.10 mg/a*
Chamber No. 2; Given Concentra-
tion 0.03 mg/m3
Variation Linits
%
Variation
Limits
%
0,07
0,09—0,11
0,12—0,15
0,8
86,9
12,3
0,028-0,033
0,034-0,040
0,041—0,050
0,050
56,0
25,4
13,8
3,8
X±Sx
0.10 i 0,007
X±Sx
0,036 ±0,003
NaF dust, which entered the exposure chambers, must be classified as
highly dispersed: particles of up to ly comprise 75.0-89.2%, and those of
l-2y, 10.8-25%. The natural content of F~ in the food and water was the
same as in the experiment with HF (the daily intake of F~ with the food was
0.045 rag, and with water, 0.007 mg).
During the first month of exposure, as in the case of HF, some of the
animals of the second group were, stimulated. The "fluorinated" and control
animals did not differ in weight over the course of the experiment.
Group I (NaF-0.10 flR/n*)
Duj;in| Period
posure
Duricg Recovery
Period •
0.015
Group n (NaF-0.03 tng/r.'
OOtO
0.005
I
| 0.0)0
I 0.005
I
1
Control
/
---i—
Observation Time, Months
1. Average data for the chronaxy of Antagonist
muscles in each group.
1 - extensors; 2 - flexors
- 36 -
-------
The functional state of the nervous system was studied by the method
of motor chronaximetry on ten animals of each group. In animals of the
first and second groups, a slight but statistically significant prolongation
of the chronaxy of the extensors and flexors was recorded. No disturbances
of the ratio of chronaxies of antagonist muscles were noted (Fig. 1). Data
from the measurement -of motor chronaxy attest to a certain inhibition of the
processes in the central nervous system, under the influence of KaF. Under
the influence of similar HF concentrations, a prolongation, equalization and
inverse ratio of the chronaxies of the antagonist muscles were observed in
the animals.
At the end of exposure of animals of the first group, morphologic
changes were observed in the cerebral cortex: deformation of dendrites,
nerve cells of enlarged volume with clarified cytoplasm and vanished Nissl
substance. In addition, there was a wrinkling of the nerve cells and
pyknosis of the nuclei. The process was focal in character. After the one-
month recovery period, altered neurons occurred much more seldom than
immediately after the exposure. An HF concentration of 0.10 rag/m^ caused
more profound destructive changes in the cerebral cortex, up to and includ-
ing the destruction of nerve cell nuclei and neuronophagia.
During exposure to NaF in a concentration of 0.03 mg/m^, the animals
of the second group showed sligiht changes of neurons which could be regarded
as functional and reversible, i.e., they were not observed one month after
the exposure.
The content of active sulfhydryl groups in the brain tissue, determined
by the Kolthoff and Harris*method, was the same in the "fluorinated" as in
the control animals.
A study of blood cholinesterase activity on eight animals of each group
using the Fleischer-Pop^'method showed that both NaF concentrations, like
HF, had an inhibiting effect on this enzyme beginning with the first month
of exposure (Fig. 2).
Sodium fluoride also has an inhibitory effect on the activity of blood
alkaline phosphatase (the phosphatase activity was determined on six animals
of each group according to G. K. Shlygin and S. Ya. Mikhlin). An NaF concen-
tration of 0.10 mg/m^ causes an inhibition of the activity of the enzyme in
the animals over the entire period of exposure. During exposure to 0.03
mg/m^ NaF, there was an obvious tendency toward a decrease in the activity of
the enzyme, but statistically reliable differences from the control occurred
only during the first two months of exposure (Fig. 3).
Having established the fact of the inhibitory effect of NaF on phospha-
tase, which is involved in the phosphorus and calcium metabolism and in the
processes of bone formation, we thought it desirable to study the phosphorus-
* [ -Irarslator's r.ote: Kcl'tgoff ar.d Garis, Fleysher and Po'jp, according to tne transliteration- of
*• R-ssisr. refersnctf. j
- 37 -
-------
calcium metabolism as well, by using the method of isotopic tracers. Upon
intraperitoneal -administration of the P^2 and Ca^5 radioisotopes, it'-was
noted that the phosphorus metabolism tended to speed up in the bone tissue
and humerus and that there was a retardation of the calcium metabolism in
the femur. After the one-month recovery period, the phosphorus-calcium
metabolism returned to normal- The content of stable P and Ca elements in
the bones, determined by spectral analysis, did not change under the in-
fluence of the NaT concentrations studied.
vuring
Recovery
Period
Daring exposure
Observation time, r.or.ths
Fig. 2. Blood cholinesterase activity or txperisental animals.
1 - NsF - 0,10 ag/n5; 2 - NaF - 0.03 cg/mJi. -3 - control.
During exposure
Before
exposure
'ft
During
Recovery
Period
I
tlVF
-JiUl£3_
Observation tioe, months
Fig. 5. Activity of blood alkaline phesuhatese of experi-
mental animals.
Notation same as in Fig. t\
- 38 -
-------
During the exposure, observations were made every month to determine
the excretion of fluorine from the body through the kidneys, and at the
end of the exposure, the deposition of fluorine in the bone tissue was
determined (fluorine was determined by ion exchange chromatography). It
was found that in the first and second groups of animals during the ex-
posure, the fluorine content in the urine was respectively 4-5 and 2-3
times that of the control group (Fig. 4). However, the fluorine that had
entered the body was not excreted, and it accumulated in the skeleton.
After the exposure, the fluorine level in the bone tissue of animals of the
first group surpassed the control level by a factor of 5-7, and that of the
second group, by a factor of 1 1/2-3 times, but it was 2-3 times lower than
during exposure to hydrogen fluoride (Table 5). There was no damage to the
tooth enamel under the influence of the NaF concentrations studied. Histo-
logical analyses of the bone tissue established certain disturbances in
animals of the first group only. Focal changes of osteocytes in the form of
a faint color and lysis of the nuclei and also an irregular and frequently
substantial deposition of lime were occasionally observed in the dense sub-
stance of the humerus, femur and pelvic bone. After the one-month recovery
period, the altered nuclei of osteocytes had almost disappeared, but an
irregular deposition of lime was observed as before.
2
3efcre exposure
During exposure
123
11 . ///
!V
Dunr.z,
Recovery
Period
.'/l Yl! Vill
Observation time, ment.is
f'.ig. 4, Fluorine content in tile urine of experimental animals,
with confidence limits.
N'otstisn same as in Fig. 2.
The state of the respiratory organs deserves some attention. In con-
trast to the experiment with HF (an irritant gas with a low pH), the upper
respiratory tract remains unchanged during inhalation of a finely dispersed
sodium fluoride dust. The latter penetrates into the deep reaches of the
respiratory organs, causing injury to the lungs at the 0.10 mg/m^ concentra-
tion. The interalveolar septa are enlarged as a result of being infiltrated
by lymphocytes, histiocytes and epithelioid cells. The capillaries of inter-
alveolar septa are expanded and overfilled with erythrocytes. Numerous fine-
focus hemorrhages and areas of desquamative alveolitis are observed. Pulmon-
ary emphysema is pronounced. Some changes in the lungs were also detected
one month after the exposure was discontinued (enlargements of the interalveolar
- 39 -
-------
Table 5
Fluorine Content Cin Milligram-Percent of Moist Substance) in Teeth, Huaerus and Feit-jr.
Group cf Animals
Teeth
Humerus
Femur
Statistical Criteria
X±Sx
X±Sx
X±Sx
First
Second
Control
52,4 ±5.41 8.06
17,5+2,4 "3,64
7,2+1.51 —
<0,001
<0,01
6 42,5 + 6,5 5,29
7 [22,4 + 2,9 4,86
7 7,9 + 0,7! —
<0,00!
<0.001
35,5±4,6
11.7x0.9
6,8± 1,4
5,97
2,58
<0,001
<0,05
septa and emphysema). The 0.03 mg/m^ NaF concentration does not cause any
serious changes in the lungs.
Histological analysis of the liver in animals of the first group
showed a varied staining of the protoplasm of the liver cells. The tissue
was mottled in appearance - clear foci with unstained cytoplasm alternated
with normally stained areas. Clear nodules from fragments of hepatic cells
and reticuloendothelial type cells were observed.. There was a lymphohistio-
cytic infiltration around the vessels. No appreciable histomorphologic
changes in the liver were found in animals of the second group.
15
10
V.
•H
8-3. '¦*
U B
10
05
0
c
. 4-J O
S r-i
O
4_> U
§£
CJ
After chronic
exposure
HI-
m
i
%
i ^ j
At end of re-
covery period
/ 3
Fig. 5» Content of SB groups in
the liver tissue of experimental
animals. Notation same as in
Fig. 2.
During both periods of observation, i.e., at the end of the exposure
and at the end of the one-month recovery period, the content of active SH
groups in the liver of "fluorinated" animals was lower than in the controls.
However, statistically reliable differences were established only in the
first group at the end of exposure (Fig. 5).
- 40 -
-------
The heart, kidneys, adrenals, and spleen were not damaged by the action
of the sodium fluoride concentrations studied.
Conclusions
1. The threshold of the reflex effect of NaF on the human body
established by the method of dark adaptation is 0.05 mg/m^.
2. NaF concentrations of 0.10 and 0.03 mg/m^ during round-the-clock
chronic exposure of the animals have a generally toxic effect and cause
the accumulation of fluorine in the bone tissue. The extent of the changes
observed in the body depends on the NaF concentration in air.
3. Because of the neutral properties of sodium fluoride dust and its
incomplete absorption in the body after penetration through the respiratory
organs, its toxic influence is less than that of similar hydrogen fluoride
concentrations.
4. The highest single NaF concentration in the air of populated areas
should not exceed 0.03 mg/m^, and the mean daily concentration should not
exceed 0.01 mg/m^.
LITERATURE CITED
Note: References mentioned in this paper ate to be found at the end of
the volume in the 1968 bibliography.
- 41 -
-------
BIOLOGICAL EFFECT OF POORLY SOLUBLE FLUORIDES
M. S. Sadilova
Sverdlovsk Institute of Labor ! .ys^iene and Occupational Diseases
From Akademiya Meditsinakikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneriiy". Red. A. Ryazanova.
Vypusk .11, Izdatel'stvo "Meditsina" Moskva, p. 26-32, (1968)
Aluminum fluoride, a typical representative of poorly soluble fluorides,
was selected for the study. The experiments were carried out with a conden-
sation aerosol of AlF^ (at a sublimation temperature of about 700°C).
To substantiate the highest single maximum permissible concentration of
AIF3, the light sensitivity of the eye was studied on three persons. An
AlFj concentration of 0.3 mg/m^ was found to cause a reliable increase of
the light sensitivity of the eye in all the subjects; a 0.1 mg/m^ concentra-
tion was found to be inactive. Data on the light sensitivity of the eye for
one of the subjects are listed in Table 1.
As in the experiment with HF and NaF, a chronic exposure of the animals
to AIF3 was carried out around the clock over the course of five months.
The concentrations of fluorine in the chambers were checked three times a
day. The withdrawal of air samples from the chambers was performed in the
same way as in the case of exposure of the animals to HF and NaF. Consider-
ing the fact that aluminum fluoride forms the hydrated complex ions AlF^"
and AIF2, which are insoluble in water and decrease the effective quantity
of the fluoride ion in the sample by 30%, in determining the AlFo concentra-
tion we introduced a correction factor of 1.33. The correction factor was
established on the basis of experiments on the determination of the fluoride
ion content in AIF3 by the method of volatilization and dissolution of AIF3
collected on a filter in a 0.2 N solution of alkali.
The animals were exposed to highly dispersed AIF3 powder: in the air
of the exposure chambers, particles up to ly comprise 67.6%; l-2y, 29.9%;
and 2-4y, 2.5%.
The experimental animals (white female rats two months old) were divided
into three groups, with 20-23 rats in each: the first group included animals
exposed to aluminum fluoride in 0.10 rag/m^ concentration, the second group to
0.03 mg/m^, and the third group was the control. The aluminum fluoride con-
centrations in the experimental chambers during the period of exposure are
listed in Table 2.
- 42 -
-------
Table 1
Light Sensitivity of the Eye During Inhalation of
Aluminum Fluoride in Subject L.
| AIF3 Concen-
tration in
mg/m
Observation Time in V.inutes*
Statis-
tical
Criteria
20
?5
30
40
Pure
Air
X±Sx
126,3±7,6
183 ±21,5
223,5± 17.9
281,9 + 31,5
10,3
X±Sx
¦ t
P
163,6±7,2
3,58
<0,05
169,2+12,7
0,55
>0,05
20S,2±8,9
0,86
>0,05
258,4 ±20,8
0,62
>0,05
0,1
X±Sx
¦ t
P
123,0± 1,7
0,42
>0,05
149,1+4,2
1,50
>0,05
190,9+17,4
1,30
<0,05
237,9 + 26,1
1,07
>0,05
llr. percent pf lS-miftL'te dark adaptation taken as 100..
Table 2
^lasiaun Fluoride Concentration (in Terms of F~)
During Exposure Period.
Chamber No. 1; Specified
Concentration, 0.10 rag/='
Chamber No. 2; Specified
Concentration 0.05 mg/m5
Variation Limits
%
Variation Limits
%
0,06-0,08
! 0,09—0,11 .
[0,12—0,10
13,8
77.7
8.5
1
!
OOO
000
PI I 1
OOOO
• •
OOO
Cl« 4- to
6,9
70,0
21,6
1.5
X + Sx 0,010 + 0,0013
X + Sx 0,036 + 0.0005C
The fluorine content of the food was the same as in the experiments
with HF and NaF. In the course of the experiment it was found that AIF3
in the concentrations studied does not affect the weight of the animals.
In animals of both experimental groups, a slight but statistically
reliable prolongation of the chronaxies of extensors was noted (by only
1 uF, compared with the control). The method of functional loading was
used to enhance the toxic effect. At the end of the exposure, each animal
was given 0.003 ml of a 40% alcohol solution per gram of body weight, but
- 43 -
-------
no statistically reliable differences were observed in the neuromuscular
excitability of the "fluorinated" and control animals. A very slight pro-
longation of the chronaxy of antagonist muscles, observed during the period
of exposure, immediately disappeared when the action of AIF3 was discontinued.
Thus, whereas serious disturbances of the subordinational influences were
found in the central nervous system, observable even during the recovery
period, less significant changes of the motor chronaxy were found during
exposure of the animals to NaF, and they were still less significant during
the action of AIF3, which also produces an inhibition effect on the blood
enzymes later than NaF does.
AIF3 has an inhibitory influence on the activity of the blood cholines-
terase, studied on eight animals of each group by the Fleischer-Pope method,
only with the 0.10 mg/m^ concentration, beginning with the end of the third
month of exposure (Fig. 1). An inhibition of the activity of alkaline phos-
phatase, determined according to G. K. Shlygin and S. Ya. Mikhlin in seven
animals per group, was observed during the fourth and fifth months of exposure
under the influence of AIF3 in concentrations of 0.10 and 0.03 mg/m^ (Fig. 2).
The mineral metabolism, studied at the end of the experiment by isotopic
tracer and spectral analysis methods, is not disturbed during exposure of the
animals to low concentrations of aluminum fluoride; tooth enamel is not
damaged either.
Before Exposure
J00
75
50
25
o
o
/
During Exposure
IS
I m
u
m
!V
VI
During
Recovery
Period
IP--
VI!
Observas. Ion Time, Months
Fi$. 1. Astivity of blood choline stern se of experimental
p.nnnls.
1 - AIF3 0.10 ng/m'; 2 - C.03 sg/m^; 3 - control
The fluorine content in the urine of animals subjected to the inhala-
tional action of aluminum fluoride during the period of exposure was higher
than in the control by a factor of 1 1/2 (Fig. 3), but it was considerably
lower than for animals in the experiment with HF and even NaF.
- 44 -
-------
.5
K 4J
S.
Q.
200
>50
tOO
50
Before
Exposure
t?3 I
iiitl
During Expose.-»
1l
//
///
IV
VJ
During
Recovery
Period
Vll
Observation Time, V.onths
Fij. 2. Activity of blood alkaline phosphatase of experimental
animals.
Notation same as in Fig. 1.
Our experiments have shown that during inhalation of HF, the excretion q
of fluorine by the body takes place mainly through the kidneys (about 90%);
during inhalational exposure to AlF^, the excretion of fluorine is greater
through the intestines (about 62%) than through the kidneys (28%). These
differences may be explained by the fact that HF is completely absorbed in
the respiratory organs, enters the blood, exerts a generally toxic influence,
accumulating partly in the bone tissue, and the remaining fluorine is ex-
creted from the body through the kidneys. AIF3 is swallowed in considerable
amounts from the upper respiratory tract and enters the stomach. Insoluble
in acid media, it passes into the intestines with the food, and thence,
judging from the solubility of AIF3 in the Ringer-Tyrode solution, may be
absorbed in the blood in small amounts only; most of the unabsorbed AIF3,
however, is excreted from the body with the stool.
'0.0
8.0
. 60
1.0
' 2.0
0
Before
Exposure
t?3
ife
During Exposure
ft
ftt,
11
/// IV
V
''Jfeco^ery
Period
tk
VI VI!
Observation Tims, Months
Fig. 3. Fluorine content in the urine of experimental animals.
Notation same as ::i Fig, 1,
Because of its poor solubility in biological media, AlF^ has a slight
cumulative capacity. The level in the bone tissue during exposure of the
animals to AIF3 is considerably lower than during exposure to HF and NaF.
After the animals were exposed to AlF^ in 0.03 mg/m^ concentration, statis-
tically reliable differences in the accumulation of fluorine (as compared
with the control) were observed only in the teeth. In tubular bones (shoulder
and hip) only a tendency toward an increase of the deposition of fluorine
- 45 -
-------
was observed (Table 3).
Fluorine: Content (inJSilligras Percent of.Moist Substance) in Teeth, Humerus and Femur.
Group of
Aftiaals
Teeth ,, 1 Humerus | Femur
Statistical Criteria
n
X i Sx
t
P
n
X ±Sx
t
P
it | A' ± Sx
!
i
t ! P
1
First j 9
. Second 9
Control | 8
22, 8 + 3,£
16.72:1.4
!1,8±0,7
2.85
3,13
<0,02 9
<0,01 t 9.
" 1 8
24,9±1,7
22,1x2,3
18,5± 1.9
2,50
2,21
<0,05
>0.03
1
9 |25,0i:2.3
.8 116.1 ±2,0
8 il4,0±l,2
1
4,23 | <0,C01
0.90 1 >0,05
0 Analysis of the individual indices shows that the fluorine content of
the bone tissue of the controls and "fluorinated" animals is frequently
the same.
At the end of the exposure, we determined the content of SH groups in
the liver tissue by the Kolthoff-Harris*method, but no reliable changes were
found.
A histological analysis of the internal organs and bone tissue of the
animals did not reveal any appreciable changes either. Slight changes in
the lungs in the form of focal enlargements of interalveolar septa were ob-
served in only a few animals of the first group (at the 0.10 mg/m^ AlF^
0 concentration).
Thus, during round-the-clock exposure of animals to concentrations of
0.10 and 0.03 mg/m^, AIF3 has only a slight generally toxic effect. The
0.03 mg/m3 aluminum fluoride concentration may be regarded as a subthreshold
value that causes no accumulation of fluorine in the body and produces no
histopathologic changes in the organs and tissues.
During inhalational action on the body, gaseous compounds of fluorine
(HF) are first in toxicity, next are fluorine salts, which are highly soluble in
biological media (NaF), and last are poorly soluble fluorides (AlF^, Na3Alf&,
CaF2).
Conclusions
•
1. The threshold of the reflex effect of fluorides poorly soluble in
biological media, established by the adaptometric method, is 0.3 mg/m^, and
the concentration of 0.03 mg/m3 during chronic exposure of the animals is the
subthreshold value.
¦a
* ^Translator's note: Kol'tgcff and Gsris, according to the transliteration cf Russian reference. ]
- 46 -
-------
2. In atmospheric air, the highest single maximum permissible con-
centration of fluorides (AIF3, Na^AlFg, and CaF2) sparingly soluble in
biological media is recommended at the level of 0.2 mg/m', and the mean
daily value, 0.03 mg/m^.
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end of the
volume in the 1968 bibliography.
- 47 -
-------
MATERIAL FOR STANDARDIZATION OF THE MAXIMUM PERMISSIBLE CONCENTRATION OF
HYDROGEN FLUORIDE IN THE AIR OF POPULATED AREAS
Candidate of Medical Sciences M. S. Sadilova
Sverdlovsk Institute of 'Work Hygiene and Occupational Pat.holoev
From Akademiya Meditsinskikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 10, Izdatel'stvo "Meditsina" Moskva, p. 186-201, (1967).
The literature in the Soviet Union and abroad contains extensive material
on the toxicology of fluorine and the clinical course of industrial fluorosis
(Ye. Ya. Girskaya, 1959, and others). The effect of fluorine in drinking
water has also been adequately studied. However, the importance of the rela-
tively low fluorine concentrations present in atmospheric air as a result of
industrial discharges has been insufficiently studied thus far.
For a number of years, we conducted studies to determine the level of
pollution of atmospheric air in the areas around the aluminum and cryolite
industries of the Urals. The zonal distribution of fluorine in air was in-
vestigated in connection with the setting up of industrial discharges at
various heights (M. S. Sadilova, 1958, 1959, 1964).
Simultaneously with the study of the air medium, an investigation was
made into the health of children residing within the effective range of
fluorine-containing industrial discharges. It was found that in children
subjected to the influence of fluorine compounds, the level of general
morbidity and diseases of respiratory organs is higher, and an affection
specific for fluorine is observed - mottling of tooth enamel and a higher
fluorine content in the urine. In children who live in regions where
aluminum plants are located, an increased deposition of fluorine in the bone
system was noted. These observations showed that fluorine in atmospheric air
should be regarded as one of the major factors in the pathology of the juvenile
population (M. S. Sadilova, 1957, 1962; A. F. Aksyuk and G. V. Bulychev, 1962).
Further growth of the aluminum, cryolite, superphosphate, and other in-
dustries requires the elucidation of questions concerning the safe concentra-
tion of fluorine in atmospheric air.
It should be noted that S. V. Miller (1955) has proposed maximum per-
missible concentrations of fluorine in the air of populated areas on the basis
of calculations. In validating the standard, Miller used the maximum per-
missible concentration of fluorine in drinking water, 1.5mg/l, assuming that
the fluorine compounds in water and air are equally toxic, that the amount
-------
of drinking water consumed is 1.5-2 1 per day, and that the amount of air
passing through the lungs la 15-20 per day; the author thus calculated
that the concentration of fluorine in inhaled air should not exceed 0.15
mg/m^. In other words, Miller considers a fluorine concentration of 0.15
rag/m^ (in terms of the ion) as the threshold value. Considering the fact
that fluorine has a significantly unfavorable effect on domestic animals
in areas around plants with fluorine-containing discharges into the atmos-
phere (because of the penetration of fluorine not only with inhaled air,
but also in large quantities with grass and vegetation), Miller recommends
a maximum permissible concentration with a 15-fold margin coefficient: a
highest single concentration of 0.03 mg/m^, and a mean daily concentration
of 0.01 mg/m3, both of which were adopted.
In our experiment, we studied the influence of several concentrations
of hydrogen fluoride, which is the most toxic compound present in industrial
discharges of plants. Further stages of the investigation will deal with the
influence of hydrogen fluoride and fluorine-containing dust (present both
separately and together). An experimental device vras used to determine the
threshold of olfactory perception of hydrogen fluoride in accordance with a
procedure adopted in the Soviet Union. The constancy of the concentrations
was checked by collecting samples of inhaled air before and after the study.
The threshold of olfactory perception was determined in 17 subjects with
a normal sense of smell. A total of 672 determinations were made, and hydrogen
fluoride concentrations from 0.22 to 0.02 mg/m^ were studied. The minimum
perceptible concentration ranged from 0.03 to 0.11 mg/m^. For the majority
of the subjects (10), it amounted to 0.03 mg/m^.
The maximum imperceptible concentration was 0.02 mg/m^ (Table 1).
Table 1
Concentrations of hydrogen fluoride studied in the determ-
ination of its odor threshold.
Number of
Subjects
Nuraber of
Observations
Concentrat
Threshold
ion. mt/m3
Subliminal
1
43
0. Ill
0,054
1
37
0,106
0,054
2
76
0,052
0,042
3
123
0,042
0,032
10
393
0,030
0,020
The next stage of our study consisted in the determination of the effect
of hydrogen fluoride on the central nervous system via the receptors of the
upper respiratory tract. We studied the change in the light sensitivity of
the eye of three persons with normal vision, using a procedure commonly
- 49 -
-------
employed In hygiene. Concentrations of 0.02, 0.03, and 0.06 mg/m^ were studied.
Data of the observations showed that the inhalation of hydrogen fluoride in a
concentration of 0,03 mg/m^ and especially 0.06 mg/m^ causes a marked increase
in the light sensitivity of the eye. A concentration of 0.02 mg/m^ as well as
inhalation of pure air did not cause any change in the light sensitivity of
the eye (the results are statistically significant). Fig. 1 shows data on the
light sensitivity of the eye for one of the subjects.
12000-
W00C-
33000-
3Q0GG -
* 2700Q-
ZiOOC-
•2 2/009-
/
I ISCOOr
c 12009
.a
9000r
JJinutes
Fig. 1. Light sensitivity of the eye in or.e of the sub-
jects d-jring inhalation of pure air (l). sr.d hydrogen
fluoride in concentrations of 0.02 ag/«* (2), 0.03 ag/m'
(3) and 0.06 ng/m' (k).
Thus, the odor threshold and threshold of the reflex effect of hydrogen
fluoride on the functional state of the cerebral cortex, determined by the
method of adaptometry, are at the same level, 0.03 mg/m^. These data make it
possible to classify hydrogen fluoride among substances with a trigeminal
effect.
A chronic round-the-clock exposure of the experimental animals was carried
out over the course of 5 months.
The animals were exposed in exposure chambers. The hydrogen fluoride was
dispensed by a special device whose glass section was coated with paraffin (in
order to prevent the glass from corroding). Distilling flasks were filled
with hydrofluoric acid, whose vapors, appropriately diluted with pure air
- 50 -
-------
(18 1/min), were supplied to the exposure chambers. The air leaving the
chambers was purified by passing through glass vessels filled with water.
Before the start of the experiment, the concentrations studied - 0.10,
0.03, and 0.01 mg/nr - were adjusted so that they were constant in the ex-
posure chambers. The work involved .-in producing different concentrations
of hydrogen fluoride proved to be extremely time-consuming, since the degree
of dilution of hydro-fluoric acid had to be determined empirically. The
initial solution taken was 40% hydrofluoric acid. It was found that in order
to produce a concentration of 0.10 mg/nr in the chamber, hydrofluoric acid
must be diluted with water in the proportion of 1:27; for a concentration of
0.03 mg/m3, 1:500; and for a concentration of 0.01 mg/m3, 1:40,000. The
samples of air from the chambers were collected in a filter holder on an ash-
less membrane filter No. 2, and then in two absorption units connected in
sequence and containing doubly distilled water. Fluorine was determined by
the alizarin-thorium procedure of S. K. Chirkov (1957). The sensitivity of
the procedure was 0.001 mg/m3. During the exposure, the concentrations of
hydrogen fluoride in chambers 1 and 2 were checked five times a day, and in
chamber 3 four times. The predetermined concentrations in the chambers
showed only insignificant fluctuations.
The experiments were performed on four groups of white female rats
with 19-22 rats in each group. Animals two months old were used.
Group I - concentration of hydrogen fluoride 0.10 i 0.002 mg/m3. Group
II - 0.03 t 0.0001 mg/m3. Group III - 0.01 t 0.00009 mg/m3. Group IV -
control.
The fluorine content in the food ration of the rats was normal: wheaten
bread 0.13 mg 7., black bread 0.36 mg 7«, cabbage 0.17 mg 7., oat grains 0.17 mg 7.,
dry matter, milk 0.14 mg/1. The animals drank boiled tap water with a fluorine
content of 0.7 mg/1. Before and after the experiment, the general state of
the animals, their weight, and the composition of peripheral blood were ob-
served. It was noted that the majority of the animals of group II during
the first month of exposure were excited (the animals bit each other and also
showed aggressiveness toward the experimenter). Subsequently they became as
quiet as the animals of the other groups. No weight changes were detected.
The composition of peripheral blood (hemoglobin, erythrocytes, reticulocytes,
leucocytes, differential leucocyte count) did not show any significant devia-
tions from the norm.
In setting up the chronic experiment, particular attention was paid to
the study of the functional state of the central nervous system of the ex-
perimental animals. We studied the state of the central nervous system by
using the methods of conditioned reflexes and chronaximetry. The method of
neurohistology was also employed. The tests were conducted on 8-11 animals
of each group.
- 5! -
-------
.In the study of the conditioned reflex activity against a background of
chronic exposure, use was made of an accelerated variant of the motor-food
method for small animals. In animals in Kotlyarevskiy's chamber, two positive
conditioned reflexes were developed - to a bell and to red light, and one
negative reflex - to a buzzer. After reinforcing the; positive reflexes, a
stereotype was developed. The stereotype included seven signals that were
alternated in a definite sequence: bell, light, light, bell, buzzer, bell,
The influence of fluorine on the conditioned reflex activity was
estimated from the rate of formation and reinforcement of the reflexes, the
number of correct responses, and the length of the latent period. In the
development of the stereotype, account was also taken of the state of the
differential inhibition, successive inhibition, and character of the inten-
sity relationships of reflexes to stimuli of different intensities. The
chronic action of hydrogen fluoride in concentrations of 0.03 and 0.01
mg/m^ was also studied against a background of functional loads.
The studies showed that hydrogen fluoride in a concentration of 0.10
mg/m^ causes serious disturbances in the conditioned reflex activity of ex-
perimental animals: a retardation of the reinforcement of positive reflexes,
an insufficient stability of the latter, and a lengthening of the latent
period (Table 2).
light.
Table 2
Formation and Stability of Positive Conditioned Reflexes in Experi-
mental Aniroals Under the Influence of Hydrogen Fluoride
Con— Statis-
cen- ticai
tra- IrrSices
tior...
Bell
Light
Appeared) J?s_ ;tage of ; appeared,
I forced '»2rf«i°gcl ' forci
ng/m*
I forced '»<.p-[«wgcl ' forced Beflexes
0,10 M + m 4.2+0,5:15.1 + 2,9 6,3+0,8 5,7+0,838,l;r7,6 17,0+1,3
t 1.6 (c)» I 3,6 (b) 5,7 (c) 4.1(c) 4.3(c) 9.8(c)
0,03 Mirm 4,3±0,3;9.5±2,3
t i 3.0(b) 2,1 (a)
5,5+1,7 3,9+0,4 11,3-4,5 6,3+1,0
2,3 (a) 2,3 (a) 1,5 (o) 4.0 (b)
0,01 M±m 3,3 = 0,14,7^1,1 1,3 + 0.4 2,4 + 0,2 4,3+1,0 1,8+0,4
t | 1,6(0) 0,2(0) 0,2 (o) 0 (o) O.i (o) ( 0.6 (o)
Control M + m 3,4 + 0,1 4,5 = 0,6 1,4 + 0.3 2,4 ' 0.2 4,5 + 0,9'2,1 -0,3
~ Degree of significance: a - 95$; b - 99%; c - 99.9#;
o - not significant
- 52 -
-------
In the development of the stereotype ir> these animals, there was recorded
a large percentage of cases of successive inhibition and disinhibition of
differentiations as compared with the groups of animals exposed to lower con-
centrations of hydrogen fluoride and with the control group. Also observed
were disturbances of the intensity relationships of conditioned reflexes to
the weak and strong stimuli (appearance of equalizing and paradoxical phases,
Table 3).
Table 3
Effect of Hydrogen Fluoride on the Conditioned Reflex Activity of
Experimental Animals in the Process of Formation and Heinforcsmer.t.
of the Organist..
Stat is-'
Concentration, mg/ir.' 1
Indices
tical
Indices
0.10
0.03
0.C1
Control
Latent period:
To bell
M + ni
r
1,37 r
0,03
16.7(c)*
0,63 ±
0,04
2.0(o)
0,69 +
0,02
1,1 (0)
0,65
0,03
To light ,
Disturbance cf
Intensity
Relationships:
M + m
t
2,09 +
0,05
I i,5 (c)
1,38 ±
0,03
0.7(e)
1,49 ±
0,03
1,8 (0)
1.41 x
0,03
Equalizing phase
M ± m
t
3,3 + 1,6
i ,3 (o)
3.0 ± 0,9
l.0(o)
Her
1,8 ± 0,7
Paradoxical pnase
M -fc m
l
6,3 + 1,40
4,5 (c)
2,7 + 0,9
3.0 (b)
Hct
HeT
Loss of reflex:
To bell
M + m
1
3,0+0,45
6.6 (c)
Hct
0,11 '
0,!
1,1 (0)
Hei
To light
Successive
Inhibition:
To bell
M + tn
1
At + m
t
13,0 L 1.2
10,4 (c)
23.3-1-
4,02
5,7 (c)
2,6 + 2,6
3.6 (b)
13,3 +
3,3
3.9 (c)
0,33 ~
0,18
0,1 (0)
0.33 -b
0.18
0,6 (0)
0,36 ±
0,12
O.H.0,13
To light
M + in
i
'31.0 + 5,1
5,9
17,7 +
4,7"
3.3 (I.)
0,77 -
0,29
0,32 (0)
;0,9 + C.3
1
* Degree of significance: a - b - 99$; c - 99.9^i
o - r.ot significant.
- 53 -
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Changes in the conditioned reflex activity were also noted in animals
subjected to the action of hydrogen fluoride in a concentration of 0.03 mg/m^,
but they were observed with particular clarity against the background of
functional loads (24-hour starvation). After the 24-hour starvation, animals
of this group showed an increase in the number of losses of conditioned re-
flexes to light and an increase in cases of successive inhibition.
A concentration of 0.01 mg/m^ caused no changes in the conditioned reflex
activity of the animals.
Toward the end of the one-month recovery period, the conditioned reflex
activity of the animals subjected to the action of hydrogen fluoride in a con-
centration of 0.10 mg/m^ improved somewhat, but remained altered relative to
the control. The conditioned reflex activity of the animals which inhaled
hydrogen fluoride in a concentration of 0.03 mg/ra^ was restored completely.
The motor chronaxy of antagonist muscles was measured on the shin of the
right hind limb. The tests were performed with a GIF condenser chronaximeter
an average of three times a month at a fixed time and under the same conditions.
The nerve cordB were stimulated by the unipolar method. Average data on the
measurement of the chronaxy of antagonist muscles according to groups are
shown in Fig. 2.
In rats of groups III and IV (control), the subordinational influences
of the brain were within the normal range.
In the experimental animals of groups I and II, changes in chronaxy were
observed immediately after the start of exposure and were manifested in a pro-
longation, equalization, and reversed ratio of the chronaxies of antagonist
muscles. It should be noted in this connection that the prolongation of the
chronaxies in animals of groups I and II was characteristic chiefly of flexors,
whereas a prolongation of the chronaxies of extensors was observed in 38.4
and 30.87. of the cases respectively. The results of the tests are statisti-
cally significant.
The changes which we observed in the activity of the central nervous
system may be regarded as a manifestation of a deep inhibition encompassing
the complex system of cortical and subcortical subordination centers. In
the experimental animals of groups I and II, there was established a statisti-
cally significant depression of the activity of cholinesterase, the enzyme
involved in the transmission of nervous excitation.
Neurohistological studies* of the brain were conducted in the area of
motor and sensory systems. Interneuronal connections and the state of the
nerve cells were studied in this case.
* I he histological examine*icns were per-crnied by 0, K, Sht:irkma (Sverdlovsk Medical -nstitutey.
- 54 -
-------
8 ,
i z j i s s 7 a
o.ow
aoos
_ . _ ^ « s e 7 e
Tine of observation (con-hs)
Fig, 2. Chronaxy of antagonist muscles in rats
during their exposure to Hydrogen fluoride in
groups I, n, HI, and IV,
1 - extensor muscles; 2 - flexor muxcles;
AB - period of exposure.
The following symptoms were observed in animals of group I: hyperemia of
vessels of the membranes and substance of the brain, perivascular edema, thick-
ening of dendrites with the formation of beaded swellings on them and the dis-
appearance of cytoplasmic outgrowths, the so-called spines. With regard to
the nerve cells, a change of their structure consisting of an uneven staining
of the Nissl substance, its partial lysis, thickening, swelling, and shriveling
of the cytoplasm were established. In some nerve cells, death of the nuclei
and neuronophagia were observed. At the end of the three-month recovery period,
some shifts tending to offset the process were noted: the destructive changes
in the nerve cells were less distinct, and the spines on the dendrites were
seen more distinctly. The process was not completely compensated, since in
some nerve cells profound destructive changes were observed, i. e., cells with
lost nuclei. These data account for the fact that the conditioned reflex
- 55 -
-------
activity of animals of group I during the recovery period continued to be
altered. For this group of animals, disturbances of the nervous activity
cannot be considered to be exclusively functional.
Some authors (De Eds, 1936; Roholm, 1937; Thomas, Wilson, and De Eds,
1935) attribute the changes in bone tissues characteristic of fluorosis to
a disturbance of the phosphorus-calcium metabolism as a result of the inhib-
itory action of fluorine on the enzyme phosphatase. These authors postulate
that phosphatase participates in processes of bone formation by liberating
inorganic phosphorus from esters of phosphoric atid; this phosphorus combined
with calcium ions forms phosphorus-calcium salts necessary for building bone
tissues. Roholm (1937) ascribes in these processes a prime importance to
alkaline phosphatase. However, literature data on the effect of fluorine on
the activity of phosphatases are very contradictory.
Our observations on alkaline phosphatase were carried out on 6 animals
of each group. The phosphatase activity was determined by using the procedure
of G. K. Shlygin and S. Ya. Mikhlin. It was found that the initial period of
increase in the activity of the enzyme in animals of groups I and II is follow-
ed by a decrease in this activity. The activity of alkaline phosphatase in
animals of group III, exposed to hydrogen fluoride in a concentration of
0.01 mg/m^, was at the level of the control.
Despite the fluctuations in the activity of alkaline phosphatase during
certain periods of exposure, the inorganic phosphorus content of the blood
was practically stable and did not exceed the limits of the physiological
norm. This is apparently because the phosphorus level in the blood is not
maintained by the activity of phosphatases alone. It is known that the
activity of phosphatases even in a healthy organism is not a constant, while
the content of inorganic phosphorus fluctuates within very narrow Units.
Probably the mechanism by which inorganic phosphorus is maintained at a
constant level in the blood is complex and varied. It has been established
that phosphoric acid salts can immediately enter the blood from their reserve
in the bones when the inorganic phosphorus content of the blood has dropped.
Since the literature contains indications of a decrease in the activity
of bone phosphatase in rats acted upon by fluorine, we also checked its
activity in our experimental animals after the end of exposure. Bone phos-
phatase was determined in the tibia of 6-11 animals of each group using A.
Bogdanskiy's procedure with slight modifications by Ye. P. Yeremin and Z. A.
Kasparskaya (1950). Our studies did not show any change in the activity of
bone phosphatase under the influence of the hydrogen fluoride concentrations
studied. As soon as clinical symptoms of fluorosis became known, it was
commonly accepted that one of its early manifestations was the mottling of
tooth enamel.
- 56 -
-------
&
Before the start of the experiment, the condition of the teeth was checked
in animals of all the groups, and was subsequently checked regularly in the
course of exposure. The tooth damage was evaluated on a 4-point scale. Our
observations (Table 4) confirmed that the mottling of tooth enamel is indeed
an early specific manifestation of morphological changes under the influence
of fluorine. In white rats, the color of healthy teeth is yellowish, and
the teeth are shiny and resemble amber. In 5 animals of group I, as early as
after the first month of exposure, the teeth became transparent, whitened, and
lost their shine. As the period of exposure grew longer, the extent of damage
increased, and toward the end of the 5th month, all the animals showed a
moderate form of damage: the upper and lower incisors resembled marble, they
were, rough, streaked, and the ends of the incisors were worn down.
In animals of group II, the changes in the teeth were less marked and
appeared later than in group I.
Teble b
.V.cttling of Tooth Enamel in Experimental Animals
Number
?f
Animals
Observed
19
20
22
Degree of Damage
lis# of Observation
During the Pgnod of
Group I
Very little
Li^le
Uoder&tely
Markedly
Group II
Very little
Little
Moderately
Markedly
Group III
Very little
Little .
Moderately
Markedly
Number of Animals
with Danaged Teeth
9 2
4 ! 2
2 1 15
2
17
19
At the end of exposure, histological analyses of animals in groups I
and II showed a thinning of the enamel and dislocation of the enamel prisms.
In the animals of these groups, definite changes in the bone tissue were also
noted. In animals of group I, a narrowing of the bone-marrow channel of tubu-
lar bones was observed, and the outlines of the periosteum were irregular. In
the pelvic bones there was disturbance to the structure of the osteons*, lysis
* Editor's .iote: probatly bone tissues.
- 57 -
-------
of osteocytes, and in the region of the periosteum, a focal resorption of the
osseous substance with growth of connective tissue. Calcium was deposited
unevenly in the bones. In animals of group II, the changes were less pro-
nounced and were localized primarily in the lumbar section of the Spinal
column and in the pelvic bones. An irregularity of the periosteum, focal
disorganization of the structure of osteons, and an irregular deposition of
calcium were noted.
The changes observed in the teeth and bone - tissue constitute an irre-
futable proof of a disturbance of the phosphorus-calcium metabolism under the
influence of hydrogen fluoride.
H. Christiani attaches considerable Importance to the level of deposition
of fluorine in the bone skeleton, assuming that an increased fluorine content
of bones constitutes a diagnostic means of detecting early symptoms of fluorine
intoxication. He also holds that the severity of osseous changes is related
to the extent of deposition of fluorine. After the completion of exposure,
we determined the deposition of fluorine in the teeth and tibia of 6 animals
of each group. The teeth and bones were carefully separated from the soft
tissues, dried to a constant weight, ground up, then a sample of 0.03 g was
taken, calcinated, and dissolved in 3 drops of lcl hydrochlpric acid, then
passed through an ion exchange column with an AV-17 anion exchanger. The
amount of fluorine evolved from the column was then determined by the alizerin
thorium procedure. The analyses^showed that hydrogen fluoride concentrations
of 0.10 and 0.03 mg/m^ cause the accumulation of fluorine in the organism.
We can draw a certain parallel between the osseous changes and the level
of deposition of fluorine in the bones, and In this respect our data are in
accord with H. Christiani's point of view.
Table 3
Fluorine Content.in tbe. Tooth arid Bone .Tissues of Experimental
Animais Cm milligram-percent cf dry substance
Group of
Animals
Teeth
M+m
1
Tibia
M+ m
1 .
II
III
Control
: 10.5*5,60
18,2+1,20
15,2 + 2,11
12.2-tO.9S
17.27 (c):
3'. 87 (b)
1,27 (o)
121 ,3*8,70
51, <1+3,20
28,9 + 2,85
23.3 + 4,0
9.57 (?)
5,12 (r)
1.11 fol
* Degree of confidence: b - 99&i c - 99-9$; o - not sign-ficsnt
During the experiment, we followed the fluorine content in the urine.
Fluorine in the urine is also determined by ion exhange'chromatography, which
was first used for this purpose in the U. S. A. (Harold, Nielsen, Logan, Utan,
1960; Talvitie, Brewer).
- 58 -
-------
An increase of the fluorine concentration in the urine was noted starting
with the 2nd month of exposure, and its highest level, which substantially
exceeded the control in all the experimental groups, was established toward
the end of exposure. Despite the high degree of excretion of fluorine, the
latter, as indicated above, was deposited in the bone system.
Histological analysis of the viscera (after the completion of exposure)
in animals of group I showed in the upper respiratory tract, desquamation of
epithelial cells with the baring of the basal layer,'hyperemia of the vessels,
and focal inflammatory infiltratiop with lymphocytes, leucocytes, and histio-
cytes. Symptoms of acute focal inflanmation of the nasal mucosa were pro-
nounced in all the animals, but in different degrees. Symptoms of bronchitis
and peribronchitis were observed in the bronchi. The vessels of the lungs
were hyperemic, and symptoms of focal pneumonia were noted in two cases out
of 5. In the liver there was protein and fatty degeneration, and in the
kidneys, protein degeneration and an increased permeability of the arterioles.
In the spleen, hyperemia of the vessels of the red pulp was noted, and in
the white pulp there was a reduction of certain lymphoid follicles and
hyperplasia of reticular cells. The walls of the central artery of the spleen
were thickened as a result of plasma infiltration. Changes in the myocardium
were characterized by the disappearance of cross striation and separation of
myofibrils.
In animals of group II, the changes were less pronounced. In some areas,
the nasal mucosa was thinned (the structure of the cells was not distinctly
visible), and in some cases, on the contrary, proliferation of the epithelium
was noted. Hypersecretion and desquamation of the epithelium was present
in the bronchi. Hyperemia was noted in the lungs and in parenchymatous organs.
In animals of group III, the histological analysis did not uncover any
changes.
Conclusions
1. The threshold of odor and the threshold of the reflex effect of
hydrogen fluoride on the human organism, established by the method of dark
adaptation, are both at the same level, 0.03 mg/m^.
2. In a five-month exposure, hydrogen fluoride concentrations of 0.10
and 0.03 mg/m^ cause a number of disturbances in the organism of warm-blooded
animals:
a) Phenomena of inhibition in the central nervous system, with the .
0.10 mg/m^ concentration causing irreversible destructive changes in the
nerve cells;
b) Change in the phosphorus-calcium metabolism;
- 59 -
-------
c) Accumulation of fluorine in the bone system;
d) Histopathological changes in the teeth, bone system, and viscera.
3. The extent of the changes observed in the organism is related to the
concentration of hydrogen fluoride In the inhaled air.
4. A hydrogen fluoride concentration of 0.01 mg/m3 causes no changes in
the organism of the experimental animals.
5. Our investigations showed that:
a) A hydrogen fluoride concentration of 0.15 mg/m^ (in terms of the
fluoride ion 0,147 mg/m^) cannot be regarded as the threshold concentration;
b) A hydrogen fluoride concentration of 0.03 mg/m^ cannot be the
maximum permissible highest single concentration.
6. The highest single concentration of hydrogen fluoride must not ex-
ceed 0.02 mg/m^, and the mean daily concentration, 0.01 mg/m^.
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end
of the volume in the 196 7 bibliography.
- 60 -
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REFLEX EFFECT ON THE HUMAN ORGANISM OF LOW CONCENTRATIONS OF ACETIC ACID
AND ACETIC ANHYDRIDE PRESENT SEPARATELY AND TOGETHER IN ATMOSPHERIC AIR
M. T. Takhirov
A. N. Sysin- Institute of General and Communal Hygiene,
Acedeny of Medical Sciences of the USSR, and Tashkent Medical Institute
"From Akademiya Meditsinakikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 11, Izdatel'stvo "Meditsina" Moskva, p. 73~9l, (1968).
Acetic acid, CH^COOH (me thanecarboxylic acid) , is a monobasic organic
acid of the fatty series. It has a pungent acid odor; it is miscible with
water and organic solvents (alcohol, ether) in all proportions, solidifies
at a low temperature, its melting point is +16.7°C., boiling point 118.1°C.,
and specific gravity at 20°C. , 1.049. Acetic anhydride, (CH^CO^O, is an
organic compound, the simplest of the carboxylic acid anhydrides, a color-
less liquid with a sharp stifling odor; its boiling point is 139.6°C,
melting point 73°C., and specific gravity at 20°C., 1.082. It dissolves in
cold water to the extent of 12% and dissolves well in alcohol and ether.
Acetic acid has been known in Europe since the ninth century, and is
widely employed as a mordant or solvent in the textile industry, in dye
works, in the production of linoleum, cellulose acetate, alkylacetates, and
in many organic syntheses (production of esters, acetates, etc.).
Acetic anhydride is used in the chemical industry for the production
of cellulose acetate and many drugs and in the manufacture of certain
explosives.
Sources of atmospheric pollution with acetic acid and acetic anhydride
may be plants that either produce or consume them in their technological
processes. The literature contains no data on acetic acid or acetic anhydride
as atmospheric pollutants, but data from sanitary studies of the air of
industrial buildings confirm the possibility of such pollution. 6
According to the data of Ye. N. Kuprits and B. S. Shender, a study of an
acetic acid plant revealed acetic acid vapors in concentrations from 62 to
120 mg/m3 in the air of a plant section. S. L. Danishevskiy (1951), in a
study of plants producing acetic acid and acetic anhydride by dehydration of
ketene, found vapors of ketene, acetic acid and acetic anhydride in total
concentrations ranging from 1 to 70 mg/m^.
High concentrations of acetic acid vapors (125-440 mg/m^) were found in
1957 by Ghiringhelli Di Fabio in the production of cellulose acetate.
G. N. Nazyrov studied the air of sections in three hydrolysis plants where
- 61 -
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acetic acid was formed as a by-product. According to his data, the highest
concentrations of acetic acid vapors in the air of a plant section amount
to 50 mg/nP. Since acetic acid and acetic anhydride in the air of plant
buildings and in atmospheric air are in the form of vapors, they enter the
human body mainly via the respiratory tract. Acetic acid vapors cause
considerable irritation of the mucous membrances of the respiratory tract
and eyes, resulting in lacrimation, cough, and restriction of breathing.
The problem of the toxic effect of acetic acid has been insufficiently
treated in the literature.
: • The effect of high concentrations of acetic acid is discussed in
the works of Flyuri and Tsernik, N. V. Lazarev, Ghiringhelli Di Fabio,
B. V. Vladykin, A. I. Shibkov, G. G. Zakharov, Jamado, M. G. Ibragimov and
Ye. Z. Lisnyanskiy, and others. The chronic action of acetic acid vapors
causes affections of the nose, nasopharynx, mouth, larynx and also conjunc-
tivitis and bronchitis among workers. The concentrations responsible range
from 62 to 125 mg/m^ (Shender, Ye. N. Kuprits, Ghiringhelli Di Fabio). The
maximum permissible concentration of acetic acid' for plant buildings adopted
in the Soviet Union is 5 mg/m3. In the USA, it is five times as high as
the concentration adopted in the USSR for acetic acid and is equal to 10 parts
per million (by volume), i.e., to 24.5 mg/m3.
Literature data on the toxicity of acetic anhydride are very limited.
The toxic effect of acetic anhydride vapors is qualitatively analogous to
that of acetic acid, but is stronger and more dangerous because of the
removal of water from the tissues (F. Flyuri, F. Tsernik, N. V. Lazarev).
The maximum permissible concentration of acetic anhydride for plant shops
has not been established.
Thus, neither in the Soviet nor in the foreign literature have we
found any studies dealing with the influence of low .concentrations of acetic
acid and acetic anhydride vapors (of the order of the maximum permissible
values for plant sections or lower) on the human and animal organism. Their
odbr threshold concentrations have also been unknown. In view of this fact,
we decided to substantiate experimentally the highest single maximum per-
missible concentrations of acetic acid and acetic anhydride present separate-
ly and together in the air of populated areas.
To determine acetic acid under experimental conditions, we used a color-
imetric method developed by Yu. V'. Dyuzheva (1960). The method involves a
preliminary transfer of acetic acid into ether and its determination with
hydroxylamine and ferric chloride. The sensitivity of the method is 5 tug per
3 ml. The method is nonspecific: the total monobasic carboxylic acids are
determined; esters interfere with the determination. However, these impuri-
ties were absent in our studies. The air was drawn at a rate of 0.5-0.6 1/rain
through two absorbers with porous plates No. 1 filled with 2 ml of alcohol.
- 62 -
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The samples may be withdrawn by using ASM silica gel with a grain size of
1-2 mm (2 g of silica gel is placed in one modified Zaytsev absorber) at a
rate of 5 £/min (M. V. Alekseyeva).
The method of determination of acetic anhydride in air was developed
by Ye. V. Deyanova (1964). The method is based on the preparation of
hydroxamic acid by the reaction of acetic anhydride with hydroxylamine
hydrochloride and involves the colorimetric determination of hydroxamic
acid with ferric chloride. The sensitivity is 0.01 mg per 3 ml (according
to the author). In accordance with the author's recommendation, we raised
the sensitivity of the method to 0.005 mg in a volume of 3 ml. Specificity:
acetic acid does not interfere with the determination. The air samples
were drawn at a rate of 0.8-1 l/min into two absorbers connected in succes-
sion and containing porous plate No. 1, filled with 4 ml of a 1% alcohol
solution of hydroxylamine. In the determination of the combined action of
acetic acid and acetic anhydride, each substance was determined separately.
Table 1
Thresholds of Olfactory Perception of Acetic
Acid_ and Acetic Anhydride.
Number of
Concentration,
mg/m5
1
ri
as w
C-IJj
»"4 OW
ri oj
¦H
a a:
"Subjects
Minimum
Perceptible
In-
per-
cep-
tible
Acetic Acid
I
3
5
10
5
6
1,57
1,52
¦ 0,92
0,61
0,70
0,00
1,22
0,92
0.81
0,60
0,60
0,51
24
69
119
2c 3
102
162
Total
30
731
Acetic Anhydride
1
2
7
6
5
1,20
1,03
0,80
0,00
0,40—0,51
1,03
0,80
0,60
0,51
0,41
24
49
195
137
122
Total
21
533
- 63 -
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In order to determine the highest single maximum permissible concen-
tration of acetic acid and acetic anhydride in atmospheric air, we used
the methods recommended by the department of sanitary protection of atmos-
pheric air. We studied the threshold of olfactory perception of acetic acid
vapors in 30 persons. Ten acetic acid concentrations ranging from 5.51 to
0.51 mg/m3 were investigated. In all, 731 observations were made. The
threshold of olfactory perception was determined for acetic anhydride in
21 persons. Eleven acetic anhydride concentrations were studied, ranging
from 4.50 to 0.41 mg/m3, and 533 determinations were made. The results are
given in Table 1.
As is evident from this table, the threshold of olfactory perception
of acetic acid in the most sensitive persons is 0.60 mg/m3, and that of
acetic anhydride, 0.49 mg/m^. Concentrations of 0.51 mg/m^ acetic acid
and 0.41 mg/m3 acetic anhydride were found to be imperceptible.
The threshold of the reflex effect of low acetic acid and acetic
anhydride concentrations on the light sensitivity of the eye was determined
with an ADM adaptometer using a common procedure. Three persons 17, 18, and
30 years participated in the study. In all, 120 observations were carried
out.
The data obtained show that the inhalation of acetic acid vapors in
a concentration of 0.60 mg/m^ causes a marked increase of the light sensi-
tivity in the 20th minute in all the subjects. A concentration of 0.48
mg/m3 caused a statistically reliable increase of the light sensitivity in
two persons. A concentration of 0.37 mg/m3 was found to be inactive in all
three subjects.
Results of adaptometric studies with acetic anhydride showed that a
concentration of 0.50 mg/m3 causes a statistically reliable change in the
course of the dark adaptation curve in all three subjects, a concentration
of 0.36 mg/m^ in only two subjects, and a concentration of 0.25 mg/m^ did
not produce any changes in the subjects.
Results of the effect of the substances studied on the light sensitivity
of the eye are presented in Table 2.
Thus, the threshold of the reflex effect of acetic acid on the functional
state of the cerebral cortex, determined by the adaptometric method, is equal
in the most sensitive individuals to 0.48 mg/m3, and the threshold of acetic
anhydride, 0.36 mg/m3. The inactive concentration for acetic acid is 0.37
mg/m^, and for acetic anhydride, 0.25 mg/m3.
The next stage of our study consisted in determining the threshold of the
effect of acetic acid and acetic anhydride vapors on the electrical activity
- 64 -
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Table 2
Light Sensitivity of Eyes in the 20th and ?5th Minutes of Adaptation During Inhalation of Acetic Acid and
Acetic Anhydride Vapors in Percent of 15th Mirfute.
Minute
Pure
Acetic Aoid Cencentrat-ion mg/m^
Pure
Acetic Anhydride Concentration
rag/ra?
Subject
of Test
Air
0,60
0,48
0,37
Air
0.50
0.36
0.25
N.
T.
A.
20-
25-
20-
23-
20-
25-
146,0
187,0
176,5
242.8
139,5
204,8
254.0(c)
225.8(c)
269.0(c)
190.8(c)
177.0(c)
184,1 (o)
246,0 fc)
192,8 (o)
217,1(c)
239.1(o)
152,0(o)
198.2(o)
149,0 (o)
193,0(o)
178,5 (o)
237.5(o)
146,0(o)
194,9(o)
144,6
190.3
173.8
235.1
145.2
208.9
223.4(c)
217.0(c)
214.7(c)
197.7(c)
174.0(c)
247.4(b)
211.6(C)
217.4(c)
206,2 (c)
225.6(b).
149,2(o)
206.9 (o)
147.5(0)
192.0 (o)
170.2 (o)
244.5 (o)
141.2(0)
204,4 (o)
Note. Confidence factor: a - 9^, b - 99$i c - 99-9^t o - unreliable.
-------
of Che brain by means of the electrocortical conditioned reflex method
described by K. A. Bushtuyeva, Ye. F. Polezhayev and A. D. Semenenko (1960).
Our observations were carried out on an eight-channel Hungarian elec-
troencephalograph (Orion Budapest). The biocurrents were recorded from the
temporal and occipital parts of both cerebral hemispheres of the subjects,
using the bipolar method. The experiment involved five persons with a
normal function of the olfactory system and a distinct alpha rhythm.
We studied the influence of inhalation of three acetic acid concen-
trations (0.36, 0.29, and 0.18 mg/m3) as well as three acetic anhydride
concentrations (0.25, 0.18, and 0.11 mg/m3) on the change of the electrical
activity of the brain. The tests were conducted four times for each con-
centration, with alternation of the test with pure air. In all, 140 obser-
vations were made.
The data obtained were treated and checked for statistical reliability.
The material of the electroencephalograms was treated in the following
manner: in each pairing of light and gas in the first ten seconds (before
the light was turned on), the duration of desynchronization was calculated
in seconds (desynchronization smaller than 40% of the amplitude of the basic
rhythm and lasting less than 0.5 second was not considered in the calcula-
tion) , both during t'ne action of pure air and in the presence of the speci-
fied concentration of acetic acid and acetic anhydride.
The statistical treatment was carried out in two variants:
In the first variant, the values of the duration of desynchronization
were summed up for all 25 pairings during the action of both pure air and
the substance studied;
In the second variant, the values of the duration of desynchronization
of the alpha rhythm were also summed up, but only if they blended with
desynchronization caused by the light, also over 25 pairings, in the presence
of both pure air and the substance studied.
Statistical treatment according to the two variants gave almost the
same results.
Data obtained by studying the influence of acetic acid vapors on the
electrical activity of the brain showed that of the five subjects tested,
the threshold of formation of the electrocortical conditioned reflex is an
acetic acid concentration of 0.36 mg/m3 for two subjects, and 0.29 mg/m3
for the remaining three (Fig. 1). The 0.18 mg/m3 concentration was found
to be inactive for all the subjects.
- 66 -
-------
: : !— 1 i
a IRII-AcA(l-))
•mi ; ! n.
Fig. 1, Electroencephalogram of subject I.. B. Conditioned reflex
desynchror.izaticn occurred during inhalation of acetic acid ir. s
concentration of ®.29 z%/m* (at the 19th peirir.^).
1 - electroencephalogram from the left temporel-occipital area;
2 - electroencephalogram from tka right temporal-occipital area;
a - mark indicating light was turned on; b - aark indicating supply
of gas.
Results of the study of the influence of acetic anhydride vapors showed
that in three subjects, the conditioned electrocortical reflex was formed at
acetic anhydride concentrations of 0.25 mg/m3; the 0.18 mg/m3 concentration
produced a conditioned reflex desynchronization in two subjects (Fig. 2).
The 0.11 mg/m^ concentration was found to be inactive for all the subjects
(Fig. 3).
The changes we obtained were statistically reliable. In attempting to
give a general evaluation of the observations made in the study of the con-
ditioned reflex effect of acetic acid and acetic anhydride on the electrical
activity of the brain, we note that the conditioned reflex (or desynchron-
ization of the alpha rhythm) was developed in the course of a single experi-
mental day, usually at the 7th-9th pairing of light and gas (acetic acid and
acetic anhydride) and was retained until the 22nd-23rd pairing, after which
it became extinguished (Fig. 4 and 5). Desynchronization of the alpha rhythm
appeared with a minimum latent period (2-5 seconds) and continued until the
light was turned on.
Thus, we have established that in the most sensitive persons, the thres-
hold of the conditioned-reflex change of the brain's electrical activity for
acetic acid lies at a level of 0.29 mg/m3 and for acetic anhydride, at 0.18
mg/ra^. The inactive concentration for acetic acid was found to be 0.18 mg/m^
and for acetic anhydride, 0.11 mg/m^.
Combined data on the odor thresholds and reflex effects of acetic acid
and acetic anhydride are listed in Table 3.
- 67 -
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~u "ir-u
llllllf1'il/ s; ll'l1"!!" "I r '-'.At" .!t c 11'fill!l
f .'.'li.i'l fii
"" ° 1 ZhII-AcAn(7)
*11 11 1l
Fig. 2. Electrcsncephalograi of subject Zh. N, Conditioned-
reflex cesjr.chrcnization occurred during inhalation of acetic
anhydride ir. a ccn cent ration of 0.15 ng/ir,J (7th pairing).
Notation sair.e as in "ig, 1. .
J If J J *J J- b u ~l L "*« "U U
fei!1,Si»11 --Is,ii.'.ain;¦11 'V.!' 1 ' <«n; :,i I'
:v vY'
' ! "
Zhi:i-Ac*n(n)
'vr-j~L
Fig. 3. Electroencephalogram of subject Zl;. N. Conditioned-
reflex desynchronization did not occur during inhalation of
acetic snnydrice in a concentration of 0.11 rr>rh~i> (11th pairing).
Notation sar.e as ir. Fig. 1.
- 68 -
-------
ots
S.0
3.0
20
3.0
1.0
Pairing
Fig. 4. Change ir. the electrical activity of the brair.. in
subject I. R. during inhalation of different concentrations
of acetic acid.
1 - pure air( 2 - C.18 mg/m 3j 3 - 0.29
- 69 -
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Table 3
Thresholds of the Effect of Acetic Acid and Acetic Anhydride on Man
Substance
Olfactory
Perception
Lijht Sensi-
tivity cf
the eye
Electro:oTt
tjcal condi-
tioned re fie;
£
O
—4
to »
G
•H
CO
+J
Si
X
.1
zc
is
-C
t-.r-
£1
i
m Q. d>
L£ o «-4
a o x>
Active
Inactive
<11
>
• ' H
O
•C
Inactive
Concentrations, ng/m^
Aeetic Acid
Acetic Anhy-
dride
0,60
0,49
0,51
0,41
0,48
0,36
0,37
0,25
0,29
0.18
0,18
0,11
0.2
0.1
*
Acevie
¦ Aoid
0,16
0,19
0,25
0,15
0,15
0,145
htiims
Acetic
Anhy-
dride
0,35
0,24
0,18
0,18
0,087
0,06
San of Fractions j>f
Effect Threshold
0,94
0,80
1.06
0,81
0,99
0,83
Sum of fractions of mximuc permissible concentration:
active - 1.62; inactive - 1.32.
On the basis of all of the above data, we propose 0.2 mg/m^ as the
highest single maximum permissible concentration of acetic acid in atmos-
pheric air, and 0.1 mg/m3 for acetic anhydride.
In recent years, the combined action of low concentrations of atmos-
pheric pollutants has been widely investigated. Studies have been made
on the combined action of sulfur dioxide and sulfuric acid aerosol
(K. A. Bushtuyeva, 1961), chlorine and hydrogen chloride (V. M. Styazhkin,
1962), carbon disulfide and hydrogen sulfide (B. K. Baykov, 196 3), carbon
disulfide, hydrogen sulfide and Dowtherm (Kh. Kh. Mannanova, 1964), acetone
and acetophenone (N. Z. Tkach, 1965), phenol and acetophenone (Yu. Ye.
Korneyev, 1965), etc. The authors showed that the character of the effect
of a mixture of two and three atmospheric pollutants on the human and
animal body is determined as a total or partial summation. In one case,
during inhalation of a mixture of hydrogen sulfide, carbon disulfide and
Dowtherm, a certain potentiation of their effect was noted. For this
reason, the next objective of our study was to investigate the combined
action of acetic acid and acetic anhydride on the human organism. The
- 70 -
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study of this combination is of major sanitary importance: the source of
3.0
. i.O
tj
s
t>
0»
in
a
Ota
a. * 0
a
Pairing
Fig. 5. Change in the electrical activity of the brain of subject
Zh. N. during inhalation of different concentrations of acetic
anhydride.
..1 !- pure air; 2 - 0.11 ng/m3; 3 - 0.18 ng/m*
study of this combination is of major sanitary importance: the source of
their combined discharge into the atmosphere are plants producing acetic
acid and acetic anhydride, and also cellulose acetate plants, whose number
and output are increasing considerably, particularly because of a growing
•consumption of cellulose acetate in the production of rayon, plastics,
synthetic lacquers and other products.
In order to determine the character of the combined action of acetic
acid and acetic anhydride, after determining the threshold of smell of each
ingredient separately, we began the determination of the odor thresholds of
acetic acid and acetic anhydride in their mixture.
Seven mixtures of different concentrations of acetic acid and acetic
anhydride were studied in 20 subjects. In all, 338 observations were made.
- 71 -
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Results of the studies are listed in Table 4.
Table ••
Odor thresholds of Acetic Acid ar.d Acetic Anhydride for Their
Combined Presence.
N-Tiber of
Subjects
Thre sholc. C or.cen-
trations ir. mg/m3
Sue cf
Fractions
Subthreshold Con-
centrations in
np/m3
Sum of
Fractions
CHjCOOll —¦ (CHjCO),
CHjCOOl I + (CHjCOlj
2
0,60-1-0,25
1,05
0,20+0,45
0,80
3
0,37+0,28
0,98
0,29+0,24
0,50
4
0,63-1-0.25
0,96
0,20+0,45
0,77
2
0,29-! 0,24
0,96
0,19+0,24
0,80
3
0, 164-0,35
0,94
0,20+0,25
0,62
3
0,40 4-0,25
0,90
0,30 !-0,?5
0,76
1
0.30 ] 0,25
0,78
0.204-0,55
0,65
2
"r—
0,60+0,25
0,71
The studies performed showed that the odor thresholds for the most
sensitive persons correspond to the following concentrations of the mixture
of acetic acid and acetic anhydride: 0.16 + 0.35 mg/m3 and 0.29 + 0.24 mg/m3.
The imperceptible concentration of acetic acid in. the mixture was 0.19 mg/m^,
and of acetic anhydride, 0.24 mg/m3.
The material obtained should be treated from the standpoint of determ-
ination of the nature of the combined effect of acetic acid and acetic anhy-
dride, i.e., it should be determined whether we are dealing with synergism,
potentiation, or antagonism. In order to obtain comparable data, the con-
centrations studied should be expressed in fractions of the threshold values
for each of them. In other words, in order to determine the character of
the combined action of these two substances, the acetic acid and acetic
anhydride concentrations studied should be split into their threshold concen-
trations for each subject and summed up. Thus one can obtain the total con-
centrations of the mixture in fractions of the individual effect threshold.
As is evident from Table 4, the odor was perceived in cases where the sum of
the relative concentrations of acetic acid and acetic anhydride was close to
unity (0.90-1.05). The odor of the mixture was imperceptible if the sum of
the relative concentrations amounted to 0.82 and lower. For only one sub-
ject was the index of the total concentration of the active mixture equal to
0.78, and that of the inactive mixture, to 0.65. She was more sensitive to
the mixture of these gases.
The next section of our study involved the determination of the effect
of low acetic acid and acetic anhydride concentrations present together on
- 72 -
-------
the reflex change of the light sensitivity of the eye. We studied the
following mixtures of acetic acid and acetic anhydride: 0.31 + 0.25 mg/m^;
0.25 + 0.18 mg/m3; and 0.15 + 0.18 mg/m3. Three persons participated in
the experiment. In all, 60 determinations were carried out.
The effect was evaluated in the same manner as in the determination of
the threshold of olfactory perception. The data obtained are listed in
Table 5 and Fig. 6.
130
>20
§ 110
O
>
• H
1/2 //
I SO
o
80
GO
c
50
>>
>
30
UO
lime ir. minutes
Fig. 6. Change of the light sensitivity of the eye in
subject T. 3. during inhalation of acetic acid ar.d acetic
anhydride.
1 - acetic acid C.15 + acetic anhydride' 0.18 mg/a';
2 - acetic acid C.25 + acetic anhydride 0.18 mg/n3;
5- acetic acid C.31 + acetic anhydride'0.25 eg/"*? ^ - P'^rt air
From Table 5 and Fig. 6 it is also evident that the reflex change of
the light sensitivity takes place in the most sensitive persons at the con-
centration of a mixture of 0.25 mg/m^ acetic acid and 0.18 mg/m3 acetic
anhydride. The concentration of a mixture, of 0.15 mg/m3 acetic acid and
0.18 rag/m3 acetic anhydride was found' to be inactive with respect to the
course of the curve of dark adaptation of the eye.
The index of total concentration of the active mixture (with respect
to the adaptometrie thresholds) is 1.06, and that of the inactive mixture,
0.81. Thus, an effect of complete summation is observed during the combined
effect of acetic acid and acetic anhydride- on the light sensitivity of the eye.
- 73 -
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Table 5
Threshold of Reflex Change of the Light :iensitivity of the Lye in
the 20th Minute During Inhalation of a Mixture of Acetic Acid and
Acetic Anhydride.
Subjects
Threshold Concen-
trations, ng/m'
Sum of
Fractions
of Adap-
tometry
Threshold
Subthreshold Cqa<
centrations, mg/mJ
Sun:
of Frac-
tions of
Adaptor
etric
Threshold
CH.COOH + (CH.CO),
CH.COOH i- (CII.CO).
M.
T.
A.
0.25+0,18 (e)
0.25+0,18 (c)
0,31+0,25 (b)
1.00
1.01
1,01
0,15+0,18 (o)
0,15-' 0,18 (c)
0,25-0,18 (o)
C .81
0,81
0,77
Note. Confidence factor: '.j - 99*, c - ':>9.9*, o - unreliable
The third stage of our study dealt with the effect of subthreshold con-
centrations of the acetic acid-acetic anhydride mixture on the electrical
activity of the brain as determined by the method of the electrocortical
conditioned reflex.
The following mixtures of vapors of acetic acid and acetic anhydride
were investigated: 0.18 mg/m3 + 0.13 mg/m3; 0.15 mg/m3 + 0.087 mg/n»3; and
0.145 mg/m3 + 0.06 mg/m3. In all, 75 observations were made. Five persons
with a distinct alpha rhythm participated in the experiment. In all the
subjects, the threshold of formation of the electrocortical conditioned
reflex was determined for acetic acid and acetic anhydride taken separately.
According to the results of the determination of the threshold of the
electrocortical conditioned reflex during inhalation of the mixture of
acetic acid and acetic anhydride vapors and according to the statistical
treatment of the data, in three out of the five subjects the threshold of
formation of the electrocortical conditioned reflex corresponded to a mix-
ture of 0.18 mg/m3 acetic acid vapors and 0.13 mg/m3 acetic anhydride vapors,
with a total concentration index of 1.02 (relative to the individual thres-
hold in the study of the isolated effect of these substances). For two
subjects, the threshold mixture consisted of 0.15 mg/m3 acetic acid vapors
and 0.087 mg/m3 acetic anhydride vapors (the sum of the relative concentra-
tions of this mixture was 0.99 for them). The subthreshold (inactive) mixture
in the electroencephalographic tests for three subjects consisted of 0.15
mg/m3 acetic acid and 0.087 mg/m3 acetic anhydride (the total concentration
index of the inactive mixture was 0.75). For the remaining two subjects,
the index of the total concentration of the inactive mixture was 0.83, for
an inactive concentration of the mixture of acetic acid - 0.145 mg/m3 and
acetic anhydride - 0.06 mg/m3.
Fig. 7 shows graphs of the change of the brain's electrical activity
during inhalation of different concentrations of a mixture of acetic acid
- 74 -
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and aceclc anhydride.
OtS
3.0
S 10.
o.
Old
° 3.0
.2 20
Pairing
Fig. 7. Change in the electrical activity of the brain of subject
Zh. N. during inhalation of different concentrations of a cixture
of acetic acid and acetic anhydride.
1 - pure air? 2 - 0.145 ~ 0.06 tog/m5; 3 - 0.15 ~ 0.C87 ng/n3
Thus, analysis of the results obtained from the determination of the
threshold of the electrocortical conditioned reflex during the combined
action of acetic acid and acetic anhydride vapors, both for the determina-
tion of the threshold of olfactory perception and of the threshold of reflex
change of the light sensitivity of the eye, indicates a complete summation
of the effect of these substances, i.e., the sum of the active concentrations
expressed in fractions of the threshold values is close to unity, and the sum
of the inactive concentration is less than unity (0.83).
In converting the threshold concentrations of the acetic acid-acetic
anhydride mixture (based on the electroencephalograph!c test) to values rela-
tive to the maximum permissible concentrations of the ingredients for isolated
action, the minimum total active concentration in fractions of the maximum
permissible concentrations of the components was found to be
0.15 mg/m3 0.087 mg/m3 . _ __ ...
0.2 .g/.3 + 0.1 »g/m3 " 0-" + 0.87 = 1.62.
- 75 -
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The maximum inactive concentration is the total concentration at a
level of:
0.145 ^ 0,06 m8/m3 = 0>?2 + Q>6() = 1>32>
0.2 mg/m3 0.1 mg/m3
It follows that for the combined presence in atmospheric air of acetic
acid and acetic anhydride vapors, their total concentration expressed in
fractions of the recommended maximum permissible concentrations of each of
the substances for isolated action should not exceed 1.3.
Conclusions
1. The threshold of olfactory perception of acetic acid in the most
sensitive persons is 0.60 mg/m3, and that of acetic anhydride, 0.49 TCg/m.3.
2. The threshold of reflex change of the light sensitivity of the eye
during inhalation of acetic acid is 0.48 mg/m3, and that of acetic anhydride,
0.36 mg/m^.
3. The threshold of formation of the electrocortical conditioned reflex
for acetic acid is 0.29 mg/m3, and that of acetic anhydride, 0.18 mg/m3.
Concentrations of 0.18 mg/m3 acetic acid and 0.11 mg/m3 acetic anhydride did
not have a reflex effect on the electrical activity of the brain.
4. The proposed highest single maximum permissible concentration of
acetic acid in atmospheric air is 0.2 mg/m3, and that of acetic anhydride,
0.1 mg/m3.
5. During the combined action of acetic acid and acetic anhydride vapors
(according to data from the determination of the threshold of smell, adaptom-
etry, and electroencephalography), an effect of complete summation takes place,
6. The threshold mixture in electroencephalographic tests consists of
acetic acid (0.15 mg/rj3) and acetic anhydride (0.087 mg/m3) with a total
concentration index of 0.99. The total concentration index of the inactive
mixture is 0.83 (acetic acid 0.145 mg/m3 and acetic anhydride 0.06 mg/m3).
7. When acetic acid and acetic anhydride vapors are jointly present in
atmospheric air, their total concentration expressed in fractions of the
adopted maximum permissible concentrations of each of the substances should
not exceed 1.3.
- 76 -
-------
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end of
the volume in the 1968 bibliography.
- 77 -
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THRESHOLD CONCENTRATIONS OF PARAFFINS IN SHORT-TERM
AND LONG-TERM INHALATION
M. L. Krasovitskaya and L. K. Malyarova
Ufa Institute of Hygiene and Occupational Diseases and Perm Medicsl Institute-
From Akademiya Meditsinakikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 11, Izdatel'stvo "Meditsina" Moskva, p. 43-50, (1968).
In connection with the rapid rate of development of petroleum refining
and petrochemical branches of industry and an ever-increasing role of large-
capacity refineries and petrochemical enterprises, a major importance is
assumed by the study of the biological effect of hydrocarbons and their
sanitary standardization in atmospheric air.
The chief atmospheric pollutants in the area of a refinery are ali-
phatic hydrocarbons (olefins and paraffins).
The present article is devoted to the study of paraffin hydrocarbons
as atmospheric pollutants.
The data available in the literature (Kochmann, 1923; Eulenberg, 1925;
Nelson et al. 1943; N. V. Lazarev, 1954; Z. Kh. Filipova, 1961) indicate a
relatively slight toxicity of paraffin hydrocarbons. Nevertheless, their
sanitary standardization is considered necessary for the following reasons.
1. In areas around petroleum refineries, paraffins are the chief
hydrocarbon pollutants. A number of studies dealing with the content of
hydrocarbons in atmospheric air around petroleum refineries have been pub-
lished (B. P. Gurinov, 1958; R. S. Gil'denskiol'd, 1958, and others).
However, a sanitary evaluation of the data obtained and further observations
are complicated by the lack of sanitary standards.
2. In areas around petroleum refinery plants, paraffin hydrocarbons
enter into the composition of organic pollutants of the atmosphere and make
up a significant part of the "sum" of hydrocarbons widely employed in sani-
tary practice. Because of the great variety of hydrocarbons in atmospheric
air, the inconstancy of their composition and varying toxicity, the concept
of a "sum" has no sanitary meaning and requires an interpretation.
Paraffins (methane hydrocarbons) are open-chain saturated compounds.
Paraffins with up to four carbon atoms are gaseous substances, those with
5 to 15 carbon atoms are liquids, and higher representatives of the series
(with C-^g) at room temperature are solids. Paraffin hydrocarbons are narcotics
- 78 -
-------
whose narcotic effect increases with the number of carbon atoms in the
chain. In the body, paraffins do not undergo chemical transformations
and do not accumulate.
In dealing with the problems of atmospheric sanitation, the hydrocar-
bons of greatest interest are the gaseoux ones and the first liquid homologs,
since they are the chief atmospheric pollutants and, because of their physi-
cal properties, spread over considerable distances.
We studied the reflex effect of butane and pentane, the most toxic lower
hydrocarbons of the series found in atmospheric air.
The thresholds of olfactory perception and thresholds of the effect on
the electrical activity of the brain were determined. The study of the
threshold of olfactory perception was made on twelve practically healthy
volunteers ranging in age from 18 to 48 years.
The experiment with every concentration was repeated three times. We
studied the concentrations of 777, 498, 355, 242 and 305 mg/m3 for butane
and 328, 257, 217 and 155 mg/m3 for pentane. In all,. 327 determinations
were made. The results are shown in Table 1.
Tabic 1
Thresholds of Olfactory Perception
of A;iphatic,Kydrccarbons (Butane,
(Per.tune).
Concentration in mg/n'
of
Maximum
Minimum
Subjects
i
Inactive
Active
Per.tane
4
155
217
6
217
257
2
257
328
Butane
9
242
305
3
305
355
Thus, it was found that the odor threshold concentration in the most
sensitive persons was 217 mg/m3 for pentane and 305 mg/m3 for butane.
The threshold of the effect on the electrical activity of the brain
was studied by the electrocortical reflex method on four subjects whose
threshold of smell had first been determined. The results are given in
Table 2.
- 79 -
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"a. Is 2
Threshold Concentrations Causing. r arsatior. of Elsc-
trocortica] Conditioned Reflex During Inhalation.
Sub,-eot
Hydrocarbon Concentrations.
zx/m?
Pentane
Butane
ISO
130
100
350
320
2S0
200
S. A.
b
-l
-I-
I. R.
+
+
—
~n
4-
—
—
V. T.
+
—
-t
-I"
—
Kh. E.
+
+
¦ "
Conventional synbols: * electrocortical reflex
forrwd; - not formed.
The minimum concentrations causing the formation of the electro-
cortical reflex during short-term inhalation lie at the imperceptible
level and are 130 mg/m^ for pentane and 280 mg/m^ for butane.
In view of the fact that hydrocarbons of the paraffin series are
constantly present in atmospheric air, we studied the character of their
combined action. The tests were conducted on three persons whose thresholds
of olfactory perception and of the isolated effect of these substances on
the electrical activity of the brain had first been determined. The effect
of the hydrocarbons was determined in the following combinations:
Combination I: maximum inactive concentration of pentane + maximum
inactive concentration of butane;
Combination II: 1/2 maximum inactive concentration of pentane + 1/2
maximum inactive concentration of butane;
Combination III: 1/2 maximum inactive concentration of pentane + .1/4
maximum inactive concentration of butane (Table 3).
Hence, all the combinations of paraffin hydrocarbons Tor which their
sum is greater than or equal to 1 (in fractions of the maximum inactive
concentration of each substance) cause a conditioned-reflex desynchroniza-
tion of the a rhythm.
With a combination of pentane and butane whose concentrations (in
fractions of the maximum inactive value) added up to less than 1, no elec-
trocortical conditioned reflex could be formed.
- 80 -
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In order to study the chronic
effect of paraffin hydrocarbons, white
male rats weighing 85-95 g were subjected
to a chronic round-the-clock exposure.
Exposure to the first liquid homolog,
pentane, was tested.
The choice of concentrations was due
to the following considerations: 30 mg/m^
is one-third the highest single (proposed
mean daily) concentration; 100 mg/m^ is at
the level of the highest single concentra-
tion; 300 mg/m^ corresponds to the maximum
permissible value for plant shops; 800
mg/m-* was taken in order to obtain a pro-
nounced effect.
The following tests were used in the
study: weight and behavior of the animals, blood pressure, chronaxy of
antagonist muscles, cholinesterase activity of the blood, and determination
of the number of blood leucocytes. Before the end of the exposure, the
adscrptive capacity of certain tissues and organs was studied by vital
staining.
The exposure was carried out for 117 days
with the exception of series IV of the experi-
ments (exposure to pentane in a concentration
of 800 mg/tn^). The exposure of animals of this
group lasted 66 days until the appearance of
distinct changes.
At the end of the experiment, some of the
animals were killed for the purpose of vital
staining of the organs and histopathological
analyses; the remaining animals were left alive
for observation for 12 days during the recovery
period. In the course of the entire exposure,
the behavior of the experimental animals was no
different from that of the intact animals. The initial weight of animals of
all groups was practically the same (the differences were statistically unre-
liable). Observations of the weight dynamics did not show any changes at sub-
sequent stages of the exposure either.
The initial blood pressure level in all the rats was in the range of the
physiological norm for this species of animals and amounted to 113-110 mm Hg
in the various groups. A statistically reliable drop of the blond pressure
level was observed after a one-month exposure in animals exposed to pentane in
Table 3
Results of Study of the Com-
bined Effect of Pentane and
Butane on the Fprratlon pf.
tw Electrocortical Conditioned
Reflex.
Sub-jest
s
P°r.:.6ne ~
Butane
S.A.
J.rt.
V.T
1 + 1
+
-1-
+
T
-h
+
lh + 1ii
—
—"
Note. Concentrations^ of
pent.ane and Tmtare given in
fractions of maximum inactive
values.
Issle ft
Pentane Concentration During
Chronic Exposure
Cor.centratiohs, dr/i
Series of
Specified
Experi-
Actual
ments
I
30.0
25,5
II
100.0
U6±2,5
III
300
332 ±7
IV
800
800 ±20,7
V
Control
- 81 -
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concentrations of 800 and 332 mg/m^. The drop of the blood pressure level
became more pronounced in subsequent stages of the exposure. Pentane in a
concentration of 116 mg/m3 also showed a hypotensive effect after a two-
month exposure. The blood pressure level in animals exposed to the action
of pentane in a concentration of 25 mg/m^ remained unchanged.
A distinct distortion of the chronaxies of the flexors and extensors
was established after two months of the experiment in animals exposed to
pentane in concentrations of 800, 332 and 116 Pentane in a concen-
tration of 25 mg/m^ did not cause any change of subordinative chronaxy
(Table 5).
Table 5
Ratio of Chronaxias of Antagonist Muscles in Experimental and
Contrcl Aninals Before and During tr.e Experiment.
Dp.te of
StiAiy
Series I
30 !Dg/m3
M x m
Scales 11
116 ^/m3
M t
Sories IT.
52 ng/m3
M ± "»
Series IV
SC^/n.5
Control
7/X 1965 r.
10/X
10/X
10/I 1966 r.
l/II
1.69 + 0,28
1,484-0,12
l,44±0.12
1.69 + 0,21
1.56 ±0,19
1 ,67 + 0,22
!,18 ± 0.22
0,75*0,08
0,45 + 0,05
0,87 ±0,11
1,45+1,12
1,14-^0,12
0,71 +0,12
0,98-.0,08
0,72-i 0,19
1,82 + 0,32
1,29+0,27
0,54+0,12
1,49-0,19
1.36 4 0,34
1,4 4 0,12
1,66 4 0,15
1,47 + 0,12
The cholinesterase activity was determined by using-a modified Fleischer-
Pop^" method. During the entire exposure, the cholinesterase activity of the
blood (in micromoles of decomposed acetylcholine) in all the experimental
animals did not differ appreciably from the .control. A statistically reliable
difference as compared with the initial level was not found in any of the
observed groups either.
The number of blood leucocytes in the experimental and control animals
during the exposure did not exceed the limits of the physiological norm and
amounted to 8825-15580.
The method of vital staining was determined by the degree of absorption
of the dye by the various organs (method of D. N. Nasonov and V. Ya. Alek-
sandrov).
Combined data on the accumulation of the vital dye (neutral red) by the
various organs are listed in Table 6.
As is evident from the above data, the degree of absorption of the vital
dye by the brain tissue in animals of series II and III of the experiments is
approximately twice (1.9-2.3) as high as in the intact animals. According to
the remaining data, the differences are insignificant.
* [Irsnsletc-r's note: Flevsher arc Poup, aissrdir.g to the transliteration of Russian reference.]
- 82 -
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Index of Accumulation of Vital Dye (in riiligraias per gr%m of
tissue Height) in Experimental and Control Animals.
Series.of 1/ 1
Expen- CConpari-
\^-ments tration
Ot^an 25*-j ,,
sg/a?;
IX
(Cor: ?en-
tration
rag/mi)
III
(.Concen-
tration
532
mg/a')
Control
I»ain
Heart
Liver
24,7 + 3,3
106+6,4
2604-50,5
67±9,0 | 53+8,5
108+7,3 jl03±4,7
247 + 48,3 I70±16
29x8,6
9,6±7,3
155+16
All the tests were carried out two weeks after the end of exposure.
Tests that revealed changes in the course of exposure established a return
to the physiological norm at the end of the recovery period.
Studies of atmospheric air in areas around petroleum refineries estab-
lished the content of the paraffin hydrocarbons (Table 7).
Table 7
Content of Aliphatic Hydrocarbons in the Atmospheric Air ol' a
Resident Ul ^rea.
Component
' SingleCor.centrations
in rag/m3
Mean Daily Concen-
trations in rag/c3
Jlaxirar.
liininum
Average
IVaxinmn
iir.imun,
Average
Sthar.e
Propane
Butane
Pentane
2.5G
12,9
* 4,05
33,6
0.58
0,40
0,32
0.52
1.4
3.05
1,88'
7,45
2,15
21
2,76
4,05
0,85 •
0,305
0,74
0,70
1 .4!
3.59
1 ,09
2,70
Conclusions
1. The lower paraffins (butane, pentane) have an olfactory effect whose
threshold for the most sensitive persons is 305 and 217 mg/m^ respectively.
2. The minimum concentrations at which the formation of the electrocor-
tical reflex is possible in the most sensitive persons are 130 mg/m3 for pen-
tane and 2 80 mg/n^ for butane. The maximum inactive concentrations were 100
and 200 mg/m3 respectively.
- 83 -
-------
3. The highest single maximum permissible concentrations of paraffins
which are proposed are below the thresholds causing olfactory perception
and reflex shifts in the most sensitive persons: 100 rag/m^ for pentane and
200 mg/m^ f0I- butane.
4. In the combined action of butane and pentane, an additive effect
takes place, established by the most sensitive test, the EEG reflex. Thus,
when these substances are jointly present in atmospheric air, the permissible
concentrations are such that when expressed in fractions of the maximum per-
missible concentration, they do not exceed a sum of 1.
5. After a prolonged exposure of the experimental animals to the action
of paraffins (pentane), the blood pressure decreases, the subordinative
chronaxy is distorted, and the adsorptive capacity of the nervous tissue
(brain) changes. The changes already take place at pentane concentrations
of 116 and 332 mg/m^.
6. Pentane in a concentration of 25 ing/m^ during chronic round-the-
clock exposure does not cause any appreciable shifts in the body of the
animals. This concentration is proposed as the mean daily maximum permis-
sible concentration of pentane in atmospheric air.
7. Petroleum refineries pollute atmospheric air with paraffins hav-
ing up to C5 carbon atoms.
8. The results of the studies demonstrate the usefulness of the concept
of the "sum" of hydrocarbons and the need for a differentiated definition of
diverse class
-------
BIOLOGICAL EFFECT AND HYGIENIC EVALUATION OF POLLUTION OF ATMOSPHERIC AIR WITH
PHTHALIC ANHYDRIDE
L. P, Slavgorodskiy
Ukrainian Scientific Research Institute of Communal Hygiene
From Akademiya Meditsinskikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 10, Izdatel'stvo "Meditsina" Moskva, p. 86-96, (1967)
Phthalic anhydride is the anhydride of ortho-benzenedicarboxylic acid.
Its boiling point is 284.5°C, and its melting point, 128-131°C. It has the
odor of bitter almonds.
Phthalic anhydride sublimes readily. Its solubility in water is 0.6 g
in 100 ml of water at 253C. When dissolved in hot water, it converts into
phthalic acid. It is soluble in carbon disulfide, alcohol, ether, benzene.
Phthalic anhydride is marketed in the form of flat, white, frequently rhombic
crystals.
Phthalic anhydride is used in various branches of industry. It is widely
employed in the production of dyes: phthaleins, rhodamines, indigo, anthrani-
lic acid, derivatives of anthraquinone, phthalocyanines. Esters of ortho-phtha-
lic acid are used as plasticizers in the production of glyptal, polyester, and
epoxy resins. Phthalic anhydride is used in the pharmaceutical and leather
industries. *
Its ability to enter into a series of characteristic reactions makes it
widely applicable in chemical laboratories.
There are several methods of preparing phthalic anhydride. The most popular
method used in industry is based on the oxidation of naphthalene in the presence
of a catalyst at 350-500"C. The preparation and use of phthalic anhydride are
associated with its pollution of the atmosphere.
The waste gases of phthalic production after purification in scrubbers are
discharged into the atmospheric air. Part of the phthalic anhydride is discharged
with ventilation gases. In the literature accessible to us, we were unable to
find any studies dealing with the presence of phthalic anhydride in the air
around industrial enterprises.
The effect of phthalic anhydride on the human and animal organism has been
studied by many investigators. Several authors (Friebel et al., 1956; L. A.
Titunov and A. A. Denisenko, 1957; Yu. K. Korotkova, 1957, 1960; S. N. Kremneva
and M. S. Tolgskaya, 1961) studied the toxicity of phthalic anhydride introduced
into the gastrointestinal tract.
- 85 -
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Kremneva and Tolgskaya (1961) administered intratracheally a 2% emulsion of
phthalic anhydride to white rats under anesthesia. Doses above 30 mg/kg were
found to be absolutely fatal. Death of the animals occurred within the next
few hours or within a day.
An inhalational exposure to dust and vapors of phthalic anhydride was
carried out by Friebal et al. (1956) on guinea pigs. The concentrations of
vapors were maintained at the level of 644.5 mg/m , and those of dust, at 8.5
mg/nP.
In such concentrations, the product irritates the conjunctiva and respira-
tory tract. An admixture of naphthoquinone and maleic acid increases the
irritant effect.
An inhalational exposure of 10 guinea pigs to phthalic anhydride dust for
30 days, 15 minutes once a day, at a concentration of 600 mg/m^, was carried
out by Korotkova (1959). The concentrations were determined gravimetrically.
During the exposure, coughing and sneezing were observed in the animals. In
three guinea pigs after the first exposures, edema of the eyelids, hyperemia
of the conjunctiva of the eyes and liquid discharges from the nose were
observed. In some cases, a weight loss was noted. Autopsy revealed a hyperem-
ia of the mucous membrane of the trachea and bronchi, adhesions in the pleural
cavities, and a moderate hyperemia of the internal organs.
Kremneva and Tolgskaya (1961) carried out a static exposure for 6 months,
6 days a week and 3 hours a day, to phthalic anhydride concentrations of 5-12
mg/m . At the end of the 6th month, the level of arterial pressure was 70% of
the normal. Eosinophilia was observed in the majority of cases over the entire
course of exposure. Concentrations of 1-2 mg/m^ were found to be inactive under
the same conditions.
It is evident that only high concentrations of phthalic anhydride during
periods of moderate duration were studied. The effect of low concentrations
in a round-the-clock exposure was not studied on the experimental animals.
The maximum permissible concentration of phthalic anhydride for industrial
buildings was adopted at a level of 1 mg/rn^, and has not been established for
atmospheric air.
The wide use of phthalic anhydride in industry, the inadequate study of
its toxic properties, and the presence of sources of atmospheric pollution
indicate that the sanitary evaluation of this compound is quite urgent. The
purpose of the present study was to give an experimental validation of the
maximum permissible concentration of phthalic anhydride in atmospheric air.
This was done by determining the odor threshold and studying the reflex
effect of phthalic anhydride on the light sensitivity of the eyes.
- 86 -
-------
Under the conditions of the experiment, the determination of phthalic
anhydride in a current of air was carried out by using a spectrophotometry
method proposed by M. D. Manita.
The threshold of olfactory perception of phthalic anhydride was determined
on 27 practically healthy persons (374 observations). The results are given
in Table 1.
Table 1
threshold of Olfactory Perception of Fftthalic Anhydride
N umber
of
Sub-'ects
I
Number of (Mir.imum Peroep-
3bservations tible Concen-
tration, jng/cK
Uwiijsuia Imper-
ceptible Con-
centration,
6
100
0,32
0,22
4
66
0.53
0,32
17
208
0,72
0,53
It is evident from Table 1 that the minimum perceptible concentrations
of phthalic anhydride in different persons ranged from 0.32 to 0.72 mg/m^.
The threshold of odor perception in the most sensitive persons was 0.32 mg/m^.
The maximum imperceptible concentration was found to be 0.22 mg/m^. The
concentrations studied had no irritant effect on the subjects.
In recent years, the study of the effect of vapors of various toxic
compounds on the light sensitivity of the eyes has been widely used for validat-
ing the highest single maximum permissible concentrations.
The effect of phthalic anhydride on the light sensitivity of the eyes was
studied on three persons by means of an ADM adaptometer, using a widely accepted
procedure. The light sensitivity was measured up to the 40th minute every 5
minutes. The effect of each concentration on the light sensitivity of the
eyes was studied no fewer than 3 times (Table 2).
Thus, a phthalic anhydride concentration of 0.96 mg/m caused significant
changes in the 20th minute of dark adaptation in all the subjects. A concentra-
tion of 0.55 mg/m was found to be inactive.
In 2 out of 3 subjects under adaptometric observations, the threshold of
odor perception was observed at a level of 0.72 mg/m , and in the third subject,
the perception threshold was not determined.
Based on what appeared to be a primarily trigeminal action of phthalic
anhydride, the data of our studies make it possible to recommend the highest
single maximum permissible concentration of phthalic anhydride in atmospheric
air at the level of 0.2 mg/m .
- 87 -
-------
±BLit '2
Results of Adaptometr ic >s
Subject
Concentrations of
Phthalic Anhydride,
tight Sensitivity
in the 20th fiinute
of Dar|^Acaptaticn
E.
Pure Air
0,50-0,55
0,96-1 ,09,
1 ,49-1,79
23 533.
21 066 (o)
30 700 (b)
45 899 (c)
P.
Pure Air
0,50—0,55
0,95-1 .09
} ,64- -1,79
41 330
39 933 (o)
59 433 (b)
77 100 (c)
P.
Pure Air
0,50—0,55
0,96—1,09
1,64—1,79
21 067'
21 333 (o)
35 367 (a)
41 633 (b)
Note. Degree of significance: a - 95$j b - 99$;
c - 99.9$; o - insignificant.
In order to validate the mean daily maximum permissible concentration of
phthalic anhydride, we studied the influence of its low concentrations in a
70-day round-the-clock inhalational exposure. The experiments were conducted
on 60 white male rats in 100 1 chambers. Each chamber was supplied with 20-23
1/min of air to which was first passed through FPP-15 filters. Group I of the
rats was exposed to phthalic anhydride in a concentration of 1.52 mg/m3, group
II, to 0.54 mg/m3, group III, to 0.18 mg/m3. The animals of group IV were the
controls. The concentrations were chosen on the basis of the following consid-
erations. The concentration of 1.32 mg/m3 was taken as the value close to the
maximum permissible concentration for the air of industrial buildings. The
concentration of 0.18 mg/m3 in the subliminal value from the standpoint of
olfactory perception and is close to our recommended highest single maximum
permissible concentration. The concentration of 0.54 mg/m was chosen as one
that is frequently encountered at a distance of 500 m from the phthalic anhy-
dride plant in the atmospheric air of residential areas.
During the chronic experiment, observations were made on the general con-
dition of the animals, their weight, motor chronaxy of antagonist muscles,
cholinesterase activity, and morphological composition of the blood. The
behavior and activity of the rats of all three groups did not differ from
those of the controls. Rats of all the groups gained weight in uniform fashion.
- 88 -
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Studies by numerous authors have shown the high sensitivity of the study
of the ratio of motor chronaxy of antagonist muscles as a method for character-
izing the functional state of the central nervous system (Yu. N. Uflyand,
1941; A. N. Magnitskiy, 1948; A. F. Makarchenko, 1956). In the establishment
of the mean daily maximum permissible concentrations of noxious substances in
atmospheric air, this index was first used and widely employed by the staff
of the department of communal hygiene of the Central Institute for Advanced
Training of Physicians. We used this test to study the effect of low concen-
trations of phthalic anhydride on the organism of the experimental animals.
The studies were carried out on an ISE-01 electronic pulse stimulator. The
chronaxy of the antagonist muscles was determined on the right hind leg in 5
rats of each group once every 10 days under the same conditions. A distur-
bance of the normal ratio of motor chronaxy of flexors and extensors in rats
of group I occurred on the 31st day of exposure an3 returned to normal two
weeks after the end of exposure. In rats of group II, the changes were less
pronounced. In the animals of group III, no such changes were observed (Fig. 1).
msec A 3
00
aw
aos
020
aos
:0 20 S! ta SO E1 v
Tine (days)
Kig, 1. Effect of phthalic anhydride cn the motor chronaxy
of antagonist muscles of rats in group T,(a); group H (b);
group III (c) and group IV (d/. Solid line refers t: ex-
tensors and dashed line to flexors; A3 - period of exposure.
- 89 -
-------
Numerous experiments established that the cholinesterase activity changes
in many pathological conditions of the organism. This method found applica-
tions in sanitary standardization practice. G. I. Solomin (1961), V. I.
Filatova (1962), D. G. Odoshashvili (1962), and V. A. Chizhikov (1963) confirm
that the cholinesterase content changes under the influence of chemical sub-
stances.
The cholinesterase activity of whole blood was stuided by a chemical method
of A. A. Pokrovskiy's (1953), modified by A. P. Martynova (1957). The cholin-
esterase activity during the experiment was evaluated from the change in the
time of decomposition of acetylcholine. The determination was made on rats of
each group once every two weeks. On the 42nd day of exposure, a depression of
cholinesterase activity occurred in animals of group I. The time of acetylcho-
line hydrolysis increased to 49 minutes. In rats of group II, statistically
significant but less pronounced changes were also observed. The time of acetyl-
choline hydrolysis for animals of group III did not differ from the control
(Fig. 2).
According to the literature data, phthalic anhydride acts on the blood-
forming system (k. Ye, Bakaleynik, 1960; S. N. Kremneva and M. S. Tolgskaya,
1961; V. S. Anatovskaya, 1961). This served as the basis for our study of
the morphological composition of the blood. The blood analysis included
determinations of the amount of hemoglobin, erythrocytes, thrombocytes, leuco-
cytes, and a differential leucocyte count. On the 70th day of exposure, we
obtained a change in the thrombocytes in animals of groups I and II. The
leucocyte count did not change appreciably. The content of hemoglobin and
erythrocytes fluctuated within normal limits.
Thus, in the course of chronic exposure it was possible to establish a
change in the ratio of the motor chronaxy of antagonist muscles and in cholin-
esterase activity, and an increase in the thrombocyte count in rats of group
I exposed to phthalic anhydride in a concentration of 1.32 mg/np. Less
pronounced changes and later ones were observed in animals of group II. In
the course of the two-week recovery period, the disturbances which occurred
returned to normal. No deviations could be established in rats of group III.
The data obtained from the chronic exposure were subjected to statistical
treatment and confirmed our conclusions. On the basis of the chronic exposure,
we propose a mean daily maximum permissible concentration at the level of the
highest single concentration, 0.2 mg/m^.
Studies of pollution of atmospheric air with emissions from the phthalic
anhydride plant were conducted by using a polarographic method (I. G. Kogan,
1961).
The samples were collected on an FPP-15 filter mounted in a special holder
and in two absorbers connected In series and containing porous plate No. 2.
The absorbing liquid was 96° ethanol.
- 90 -
-------
10
c
e es
i so
in
¦o iff
o
l"
£ If
-
I
£
-
•
\
/
\
\
\ \
-
-
' i.
-
1 I 1 1
rl,
es
1 ¦
Background 1 2 3 & 5 S
Study
2. Effect of phthalic annydride or. -..he cr.olin-
;erase activity of the blood.
- group^I; 2 - group II; 3 - group III; 4 - group IV
vcontrol;-, AB - period of exposure
To determine the possibility of the spread of vapors and condensation
aerosol of phthalic anhydride in the atmosphere, we conducted studies around
plants producing phthalic anhydride. The output of the plants was over 10,000
tons of phthalic anhydride per year. Data on the degree of pollution with va-
pors and condensation aerosol of phthalic anhydride are present in Table 3.
From the latter it is evident that only at a distance of 1000 m were the
concentrations obtained lower than our proposed maximum permissible concentra-
tion (0.2 mg/m^).
Table J
Foliation of atmospheric air with pnthalic ar.hycride in the arsa
of the chemical plant complex on '.he leeward sice.
DistEr.ce from
Source of Dis-
charge, m
250
500
1 000
I 500
Number
of
Collected
Samples
Number of
Samples
Above the
Sensitivity
Ll^lt of
—Beth3d
Kaximua
Average
wpr.centra- ,j Con centra-
tior.s, mg/nJj tior.s, mg/m-
17
37
32
10
9
22
17
3
0,134
0,652
0,1510
0, o.>~>
0,031
0.061
0.084
0.017
A sanitary protective zone of no less than 1000 m must be provided for
this plant. The sanitary protective zone (500 m) adopted in accordance with
the standards of the building code for enterprises with an output of over
10,000 t must be considered inadequate.
- 91 -
-------
Conclusions
1. The threshold of olfactory perception of phthalic anhydride for the
most sensitive persons is 0.32 mg/m^, and the subliminal concentration is
0.22 mg/ra^.
2. A study of the curve of dark adaptation during short-term inhalation
of phthalic anhydride indicates that the threshold at which it is affected is
0.92 mg/rtP, and the subthreshold concentration is 0.55 rag/m-*.
3. The highest single maximum permissible concentration which must be
recommended is 0.2 mg/m^.
4. Chronic round-the-clock exposure to vapors and condensation aerosol
of phthalic anhydride in concentrations of 1.32 and 0.54 mg/nt^ for 70 days
causes significant changes in the ratio of chronaxies of flexors and extensors,
changes in the activity of whole blood cholinesterase, and an increase in the
thrombocyte count. A concentration of 0.2 mg/nr* was found to be inactive and
may be recommended as the mean daily maximum permissible concentration.
5. The atmospheric air around phthalic anhydride plants with a capacity
of 10,000 tons per year is polluted. In the presence of purification equip-
ment, phthalic anhydride is observed in concentrations below our recommended
maximum permissible value only at a distance of 1,000 m. For this reason,
the sanitary protective zone for this plant should be no less than 1000 m.
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end of
the volume in the 1967 bibliography.
- 92 -
-------
DATA FOR A SANITARY ASSESSMENT OF METHANOL IN ATMOSPHERIC AIR
Candidate of Medical Sciences^R. Ubaydullayev
A. N. Sysin Institute 9f General and Coimnunsl Hygiene of the Acadesy of Mrdical Ssiences of the USSR,
and Uzbek Scientific Research Institute of Hygiene, Sanitation, nnd Occupational Diseases
From Akademiya Meditsinakikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 10, Izdatel'stvo "Meditsina" Moskva, p. 65-74, (1967).
At the present time, methanol is obtained in huge amounts by synthesis,
and also by hydrolysis of plant raw material. Methanol finds extensive
applications in the manufacture of organic dyes, chemicopharmaceutical prep-
arations, formaldehyde, and other chemical compounds.
The chief sources of pollution of atmospheric air with methanol are
paint-and-varnish plants, wood-processing plants, metal-working shops,
furniture factories, and many other enterprises.
In the nature of its action, methanol is a nerve and vascular poison
with a marked cumulative property.
L. I. Kaza (1925) and V. M. Rozhkova (1948) held that an essential
role in the mechanism of action of methanol is played by the primary pro-
duct of its oxidation in the organism - formaldehyde, which paralyzes
cellular respiration and inhibits the oxidation processes.
The action of low methanol concentrations on the human and animal
organisms was first studied by Chao Cheng-ch'i (1959). The author experi-
mentally determined the thresholds of the odor of methanol (4.1 ng/m^) and
of its reflex effect on the light sensitivity of the eye (3.3 mg/m^), and
also carried out a chronic exposure of white rats to methanol vapor in
concentrations of 50 and 1.77 mg/m^ in the course of three months for 12
hours a day.
On the basis of these studies, the highest single and mean daily
maximum permissible concentrations of methanol in atmospheric air were es-
tablished at 1.5 and 0.5 mg/m3 respectively.
However, in his investigations, this author did not use the most sensi-
tive method, which is the study of the electrical activity of the cerebral
cortex for the purpose of validating the highest single maximum permissible
concentration. Moreover, a continuous, round-the-clock exposure was not
carried out in the chronic experiment. Therefore, we decided to refine
these norms by using the most modern experimental methods.
To determine methanol in atmospheric air, we used the method of
M. V. Alekseyeva (1963).
- 93 -
-------
We began Che study of the problem by determining the odor threshold of
methanol in 25 persons aged 18 to 40 years. Nine concentrations from 12.2 to
3.9 rag/m^ were tested (Table 1).
Table 1
Results of Determination of the Threshold
of Olfactory Perception of Methanol Vapor
Number
of
Subject.
Minimum Percep- i Maximum Inper-
tible Concentra- i cep^ible Conceq
: tion, mg/m3 | tration, ng/m-
10,3
8.4
7.5
6.5
5.6
4,5
9.7
7,5
6.5
5.6
4.5
3,9
As is evident from Table 1, the odor threshold in subjects with different
sensitivities ranged from 10.3 to 4.5 mg/m . The threshold concentration in
the most sensitive persons was found to be 4.5 mg/m^, and the maximum impercepti-
ble concentration, 3.9 mg/nH. Thus, our data almost coincided with those of
Chao Cheng-ch1i (4.1 mg/m ). We then studied the effect of low concentrations
of methanol on the light sensitivity of the eye by means of an ADM adaptometer
on three subjects ranging in age from 18 to 25 years. The study was made daily
at a strictly defined time once a day under identical conditions for each
subject.
A physiological background was developed in each subject, i.«, a standard
curve of dark adaptation, during inhalation of pure air between the 15th and
20th minute for 8 days. Concentrations of 4.11, 3.53, and 3.06 mg/rn^ were
tested. The 4.11 mg/m^ methanol concentration caused a marked change of the
light sensitivity of the eye in all the subjects, and the 3.53 mg/rir concentra-
tion did so in only one subject. For the latter, the 3.06 mg/nP concentration
proved to be inactive (Fig. 1).
The effect of low methanol concentrations on the electrical activity of
the cerebral cortex was studied on the 6 persons with the lowest odor threshold
(Table 2) by the method of A. D. Semenenko (1963) with the aid of an 8-channel
electroencephalograph. The test was conducted no fewer than 4 times with each
concentration and was regularly alternated with pure air.
The studies showed that methanol in a concentration of 1.46 mg/m^ affected
the magnitude of the amplitude ot the alpha rhythm and caused statistically
significant changes in all 6 subjects, whereas a concentration of 1.17 mg/m
did so in only two subjects.
The inactive concentration for all the subjects was 1.01 mg/m^. The re-
sults of the studies are summarized in Table 3. They show that the highest
single maximum permissible concentration of methanol established earlier at
1.5 mg/nP should be reduced to 1 mg/m .
- 94 -
-------
1S0 000
uoooo
^ >20000
2 too 000
Z BO 000
\r.
!' eoooo
| id ooo
J to 000
10 15 20 25 30 40
lime, sin.
Fig, l4 Changes in the light sensitivity of
the eyes during inhalation of nethanol vapor.
1 - pare airjT2 - concentration 4.11 ing/m';
3 - 3.53 3g/m^i 4 - 3.06 ng/m'
Table 2
Results of Study of the Threshold of the Reflex Effect
of Methanol Vapor on the Electrical Act ivity of the
Cerebral Cortex,
Subject
Methanol Concentration.
1.46
1.1*
1.01
G. K.
+
.i-
R. K.
-)-
—
T. A.
' —l_
Not Studied
M. K.
•4-
—
L. T.
+
l. a.
-p
Not Studied
Note. + statistically significant changes,
- insignificant changes.
To check the mean daily maximum permissible concentration of methanol, a
round-the-clock exposure of 45 white male rats weighing 100-120 g (three groups
of 15 animals each) was carried out in the course of 90 days; group I was ex-
posed to methanol vapors in a concentration of 5 mg/m3, which slightly exceeded
the odor threshold, group II to 0.5 mg/m3, which was the existing mean daily
maximum permissible concentration, and group III was the control.
During the entire period of exposure, the air temperature and methanol
concentration in each chamber were measured daily. In group I, the average
concentration was 5.31 - 0.62 mg/m3, and in group IX, 0.57 mg/nt3 * 0.59 mg/m3.
Xn the course of chronic exposure, the animals of all the groups were
healthy, active, and gained weight moderately. However, toward the end of the
period of exposure, a slight lag was noted in the weight of animals of the control
group.
- 95 -
-------
Table 3
Results of Study, of the Odor Threshold and Reflex Effect of Methanol
on the Human OrgEniso
3ata of Chao
Chfine-fth'&Z Our Data
C
concentrations, rg/m i>
Threshold
Minimum
Active
'Maximum
Inactive
Minimum
Active
Maximum -
Inactive
Olfactory Perception
4,1
3,
i
!
4,5
3,9
Light Sensitivity
of the Eye
3,3
! 2,4
i
3.53 |
3,06
Electrical Activity
of Cerebral Cortex
Not studied
I
i
1.17 ¦]
1,01
The study of the motor chronaxy of antagonist muscles was carried out on
5 rats in each group once every 10 days. The results show that a prolonged
inhalation of the methanol vapors in a concentration of 5.3 mg/m in rats of
group I starting with the 6th week of exposure causes statistically signifi-
cant changes in the chronaxy of antagonist muscles with the appearance of a
reversed ratio of their indices. Toward the end of the recovery period, the
ratio of the chronaxy of flexors and extensors returned to normal.
23/tV 3/v >3 22 P S/vi >9: 29 9/vn 13 29 n/vm 23 i/u
Dates of study
Fig. 2. Change ir. tt:e ratio of cnronaxy of antagonist zuscles
ir. rats. -Juring ir.halaticn of methanol vapors.
A ar.d fi - period of exposure; 1 - pure air; 2 - concentration
5.31 isg/oJ; 3 - concentration C.57 lag/m'-
- 96 -
-------
No changes in the chronaxy indices occured in rats of group II (Fig. 2).
Several authors (M. I. Gusev, 1960; K, A. Bushtuyeva, 1964; B. M. Mukhitov,
1961, and others) observed changes in the excretion of coproporphyrin with
the urine in animals exposed to a prolonged action of certain noxious chemical
compounds. We also used this method in our studies. The daily portion of
urine from each group of animals (5 rats in each) was collected in special re-
ceiver chambers made of glass. The extraction of porphyrins from the urine
was carried out by Fischer's method, and coproporphyrin was determined quanti-
tatively on an SF-4 spectrophotometer in the wavelength range of 400-410
The coproporphyrin was determined once every two weeks. In rats of group I
(5.31 mg/m ), the excretion of coproporphyrin with the urine per 100 g of
weight starting with the 7th week of exposure decreased sharply and remained
at this level until the end of exposure (Fig. 3). Normalization occurred after
20 days of the recovery period. In rats of group II (0.57 mg/rn^), no signifi-
cant changes were observed.
— - 3
° OS
Exposure
27/IV 4/f ti/U 2S/V 8/V! !2fW S/tl! 191*11 J/Mt H/\/m 25/m 5//X
Dates of study
Fig. 3. Excretion of coproporphyrin with the urine in rats
during inhalation of methanol vapors.
Notation sase as ir. Fig. ?.
We studied the activity of whole blood cholinesterase by using A. A.
Pokrovskiy's method (1953) modified by A. P. Martynova (1957).
The cholinesterase activity was determined in 5 rats of each group twice
a month. Before the exposure, the original levels of acetylcholine hydrolysis
amounted to an average of 38-39 minutes. In rats of group I, starting with
the 6th week of exposure, the time of hydrolysis increased to 41 minutes, i.e.,
a decrease in cholinesterase activity was noted. At the end of exposure,
- 97 -
-------
it amounted to 43 minutes (Fig. 4), and returned to normal at the end of the
recovery period. In rats of group II, no significant changes were observed.
Minutes . .
•
? B
-
—
y""~
/
/
i
—} •
-
Exposure
i i . i i r j
¦
x 28/IV 13/V ?l/v 7/vi 2t/V! 5/vn 19/vn 2/vm 12/vl/t 1/lK
Dates of sti;dy
Fig. t. Cnange in the activity of who'e blood eholi.nesterase
in rats during inhalation of methanol vapors.
Notation same as in Fig. 2.
The literature contains many papers indicating a change in the total amount
of blood protein or in its individual fractions in various diseases and during
the action of certain chemical compounds on the organism (R. Ubaydullayev,
1961; V. A. Chizhikov, 1964; P. G. Tkachev, 1964; Granati and Sekavo, 1956;
Mario and Carlo, 1957).
In a chronic experiment, we studied the effect of low methanol concentra-
tions on the protein fractions of the blood serum of rats by using paper
electrophoresis. The protein fractions of 5 rats of each group were studied.
The blooed was taken from the tail of an empy stomach every 15 days.
In group I, changes appeared 7 weeks after the start of exposure. This
was associated with a decrease in the content of albumins and an increase in
the amount of gamma globulins and beta globulins. There was no change in alpha
globulins as compared with the control. In rats of group II, the fractional
composition of the proteins remained unchanged (Table 4).
No changes were observed in the total proteins of the rats. Results of
the study show that a continuous exposure of white rats to methanol vapors
in the course of 90 days caused changes in animals of group I only (5.31 mg/m^).
The 0.57 mg/nr* concentration was found to be inactive, On this basis,
we consider it possible to recommend 0.5 mg/rn^ as the mean daily maximum
permissible concentration.
- 98 -
-------
Table !(
Total proteins and protein fractions of the blood seruo during
chronic exposure of white rats to methanol vapors.
Globulins
Period of
Exposure
Group
Total
Pro-
teins
AI bu-
mins
Alpha
Beta
Gamma
Albumin-
Globulin
Ratio
Before
exposure
I
II
III
(Control!
9,02
9,31
8,91
41,8
44,8
45,1
24,6
23.2
23.3
20,7
19,4
19,2
12,9
12,6
12,4
0,72
0,81
0.83
On 45th day
of exposure
I
II
HI
(Control)
8.79
9.01
9.02
34,7
44.1
43,6
24,0
21,5
20,9
24,0
20,2
20,9
11,3
14,2
14,6
0,53
0,79
0,87
On 90th day
of exposure
I
II
III
(Control)
9,17
9,16
8,89
32.8
42,0
42.9
23,3
21,8
21,8
25.8
21.9
21,2
18,t
14,3
14,1
0,49
0,72
0,76
On 20th day '
of recovery
period
I
II
III
(Control)
9,27
9,05
8,79
39,4
42,2
44,2
25,3
20,2
20,2
21,2
22.1
22,9
14,1
15,5
12,7
0.65
0,73
0,80
Table 5
Pol lutior., of atcospheric air with methanol vapors around the hydrol-
• ysis plants of Uzbekistan,
Distance
From
Source of
Discharge
Number
of
Samples
Single Concentrations,
Number of
with Cone
Samples
»rt ration,
tn7!
Maximum
Miniram
1 and
k bove
Jnder 1
Andizhan Plant
100 :
16
2,26
0,46
10
6
200 i
18
1,67
0,11
5
13
300
27
0.33
0,05
—
27
500 j
18
0.14 .
—
18
Fergana P
lant
100 ' :
17
4,49
0,89
16
1
200
15
2,2
0,1
9
6
300 !
25
0,83
0,11
—
25
500 |
19
0,3
—
—
19
Yengiyul'
Plant
100 1
14
1,15
0,1
,3
11
200 . 1
24
0,55-
1
—
24
300 1
14
0,05
—
— •
14
- 99 -
-------
In order to make a sanitary evaluation of the pollution of atmospheric
air around the Andizhan, Fergana and Vangiynl' hydrolysis plants of Uzbekistan,
we studied the range of pollution of atmospheric air with methanol vapors.
The tests were taken in May and August 1962 at distances from 100 to 500 m
oh the leeward side of the source of the discharge (Table 5).
It is evident from Table 5 that at distances of 100 and 200 m from the
andizhan and Fergana plants, the methanol concentrations exceeded the recom-
mended maximum permissible concentration 1 mg/nP). At the Yangiyul' plant,
this excess was observed only at a distance of 100 m. At larger distances
from the plants, all the concentrations were below 1 mg/nr*.
Conclusions
1. Atmospheric air around hydrolysis plants producing methanol is
polluted by its vapors at a distance of up to 200 m.
2. A study of the effect of methanol on the human organism showed that
in the most sensitive persons, the threshold of olfactory perception of
methanol is 4,5 mg/m^, the threshold of reflex change of the light sensitivity
of the eye is 3.35 mg/nP, and the threshold of action on the electrical acti-
vity of the brain is 1.17 mg/m .
3. The highest single maximum permissible concentration of methanol in
atmospheric air should be no higher than 1 mg/nP.
4. Chronic round-the-clock exposure to methanol in a concentration of
5.3 mg/m^ over the course of 90 days caused changes in the normal ratio of
chronaxy of antagonist muscles, activity of whol£ blood cholinesterase,
excretion of coproporphyrin with the urine, and protein fractions of the blood
serum in the experimental rats.
A concentration of 0.57 mg/m^ had no effect on the rat organism.
5. The mean daily maximum permissible concentration of methanol which
can be recommended is 0,5 mg/m^.
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end of
the volume in the 196 7 bibliography.
- 100 -
-------
DATA FOR THE VALIDATION OF THE MAXIMUM PERMISSIBLE CONCENTRATION
OF AMMONIA IN ATMOSPHERIC AIR
Aspirant (graduate student) M. M. Sayfutdinov
Uoscow Scientific Research Institute of Hygiene in. F. F. Erisman
From Akademiya Meditsinskikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V. A. Ryazanova.
Vypusk 10, Izdatel'stvo "Meditsina" Moskva,^p. 108-122*,s(1967) .
Ammonia is the simplest compound of nitrogen and hydrogen, a colorless
gas with a sharp suffocating odor and a pungent taste. Under pressure or on
cooling, it passes to the liquid state. It is soluble in ether, alcohol, and
fats. It dissolves readily and in large quantities in water, forming ammonium
hydroxide. It reacts actively with acids and other compounds. Ammonia is
obtained chiefly from nitrogen and hydrogen (I. D. Fotonich, 1965; S. A, Beskov,
1962).
Ammonia is a valuable nitrogen-containing liquid fertilizer and also the
main raw material in the production of nitric acid and ammonium fertilizers.
It is used in considerable amounts in the manufacture of ammonium hydroxide
and ammonium chloride. Ammonia is widely used in refrigeration engineering,
in the production of soda, and in nitriding steel products.
The chief sources of discharge of ammonia into atmospheric air are
nitrogen fertilizer plants, enterprises producing nitric acid and ammonium
salts, refrigeration units, coking and leather plants, and livestock breeding
farms.
The largest of these sources are nitrogen fertilizer plants and coking
sections of ferrous metallurgical enterprises. Small amounts of ammonia
reach the air from the soil and from open water reservoirs.
However, the literature available to us gives scant data on the quanti-
tative content of ammonia either in the air of industrial buildings or in
the atmospheric air of populated areas.
According to the data of V. A. Ryazanov (1961), at the site of one of
the largest chemical plants around the ammonia synthesis section, single
concentrations from 20.1 to 57 mg/m-* were observed at a distance of 10 tn,
and from 0.1 to 0.2 mg/m^ at a distance of 1 km. The mean daily concentra-
tions at different points of the plant site ranged from 0.216 to 3.309 mg/m^.
In the air of the town located at a distance of 2 km from this chemical plant,
the ammonia concentrations did not exceed 0.004-0.005 mg/m^.
- 101 -
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To study the degree of .pollution of atmospheric air with ammonia around
metallurgical plants, the Novolipetak Metallurgical Plant, the largest source
of discharge of ammonia into atmospheric air, was investigated. The coking
and nitrogen fertilizer sections are located on the territory of the plant.
In the coking section, ammonia is present in the composition of the coke-oven
gas and of the tar water. Ammonia reaches the atmospheric air as a result
of the discharge of coke-oven gas at vatious points of the plant through
leaks in gas lines and units (scrubbers, exhausters). The gases are also
emitted from open reservoirs (storage tanks, measuring tanks) where ammonia
water or tar water is stored.
The nitrogen fertilizer plant synthesizes ammonia from coke-oven gas
supplied by the coking plant. The entire technological process of this plant
takes place in a closed sy9tem; the discharge of the gas into atmospheric air
takes place during repairs, during'purging of the system and dispensing of the
finished liquid ammonia into tanks, and also during distribution among the
consumers.
We studied the atmospheric air around the Novolipetsk Metallurgical Plant
in September-October 1963 and in May-June 1964 and 1965. The latter study was
made in connection with the start-up of a new nitrogen fertilizer plant synthe-
sizing liquid ammonia. The air samples were taken on the leeward side of the
emission source at a level of 1.5 m above ground. A total of 400 air samples
were taken.
Results of the studies are given in Table 1.
As is evident from Table 1, the highest concentrations of ammonia were
found at distances of 500 and 1000 m. In 1965, as a result of the starting
of a nitrogen fertilizer plant synthesizing ammonia, the concentration of the
latter at close distances increased even more.
Tstle 1
Pollution of Atmospheric Air with Ammcnis Around the
Novolipetsk Metallurgical Plant.
Ammonia
1 i
s
i*j
Distance
I9<>3 r.
1964
l
1965
r-
Fror.
' i
Maximum
JCaxixjit
Average
y.axinrjc
Average
Average
500
] ,40
0,31
3.60
0,62
4,68
2.19
1 000
3,5
0,5
0.39
0,18
1 ,73
0.91
3 000
2,25
0.15
1,30
0.26
0,39
0.21
5 000
1.0
0,16
1.30
0,18
0,28
0,22
7 000
0,20
0.09
1 ,20
0,19
0,17
0,16
10 0U0
0,13
0,12
- 102 -
-------
A number of authors studied the pollution of atmospheric air around more
minor sources of discharge of ammonia.
Z. D. Markova (1941) detected ammonia in concentrations of 0.02 to
0.05 itig/m3 in the residential areas of Rostov-on-Don, whereas in the city
parks, the concentration did not exceed 0.02 mg/m^.
V. A. Kononova and V. B. Aksenova (1963) found ammonia in amounts from
0.015 to 0.057 mg/m^ in the air around livestock farms in a zone of up to
150 m.
According to the data of American authors, in areas with-pure air, the
concentration of ammonia in Chicago ranged from 0.0058 to 0,0143 mg/m^ (cited
by V. A. Ryazanov, 1961). According to the data of Stoklaz, the concentration
of ammonia in pure air varies from 0.02 to 0.04 mg/m^, which according to
Ryazanov (1961) is exaggerated. Thus, from the literature data cited it i9
evident that, depending on the source of. pollution, the concentration of
ammonia in atmospheric air ranges from 0.015 to 0.057 mg/m^, and in air where
special pollution sources are absent, the content of ammonia ranges from
0.003 to 0.005 mg/m^.
Ammonia is an irritating gas which affects primarily the mucous membranes
of the respiratory tract and the central nervous system. This effect of
ammonia is due to its high solubility on the moist surfaces of mucous membrar.es
and its ready absorption into the blood stream.
Cases have been described involving an acute poisoning with ammonia as
a result of the rupture of cylinders during production or when pouring
ammonium hydroxide in'everyday use (R. N. Vol fovskaya and G. N. Davydova,
1945; Ye. I. Lyublina, 1948; V. K. Trutnev and N. V. Velikorussova, 1955;
K. V. Yegorov, 1959, and others).
Vol'fovskaya and Davydova (1945), Trutnev, Lehmann (1886-1889), Horvath
(1926, 1929) and others classify ammonia among suffocating poisons with a
marked inflammatory-necrotic effect. Vol'fovskaya and Davydova also admit
the possibility of a resorptive effect.
Injury to the nervous system is manifested in the loss of consciousness
and a strong excitation to the point of violent delirium.
According to the observations of I. P. Pavlov (1896), ammonia in a dose
of 50 mg per kg of animal weight caused drosiness and ataxia, and when the
dose was increased by a factor of 2%, convulsions followed by death of the
experimental animals were observedi Studies made by V. V. Pravdich-Meminskiy
(1958), Ye. A. Vladimirova (1938), E. E. Xosyakov (1962), N. B. Kozlov (1962),
E. E. Martinson (1962) and others confirmed Pavlov's observations.
- 103 -
-------
Studies made by Recine (1956) showed that in poisoning with ammonia,
because of its action on prothrombin, the coagulability of the blood is
impaired, and the content of residual nitrogen in the blood increases.
An increased content of ammonia in the blood and tissues is accompanied
by changes in the physicochemical properties and structure of the tissue
proteins, a disturbance of the ion exchange and acetylcholine exchange, and
a depression of tissue respiration (N. B. Kozlov, 1962).
The chronic action of gaseous ammonia in low concentrations is manifested
in the form of inflammation of the upper respiratory tract, conjunctivitis,
and a lowering of the resistance of the organism to infectious diseases.
Workers of the municipal sewer network, who because of the requirements
of their work are exposed" for long periods of time to an atmosphere contain-
ing small amounts of ammonia and hydrogen sulfide, have displayed chronic
hypertrophic catarrhs changing into atrophic ones (M. D. Ayzenberg, 1927;
T. V. Ass, V. V. Vol et al., 1926).
Abadie, Trachta, and Trousseau (cited by R. M. Zhmudskaya, 1933) re-
vealed conjunctivitis involving ulcerations of the cornea in workers exposed
for a long time to the action of low concentrations of ammonia.
According to the data of Henderson and Haggard (1930), the threshold of
perception of ammonia for man is 37 mg/m^. According to I. M. Alpatov (1964),
the threshold of the reflex'effect of ammonia for man is 22 mg/rn^. According
to Lehman (1886), Silverman, Whittenberger, and Muller (1949), the highest
permissible concentration of ammonia for lengthy exposure is 69mg/m^. The
maximum permissible concentration of ammonia for the air of industrial build-
ings, adopted in 1930 in accordance with a proposal of the Moscow Institute
of Labor Protection, is 20 mg/nP.
Despite a considerable number of sources of discharge, ammonia as an
atmospheric pollutant has been inadequately studied. The existing studies
give contradictory data. The maximum permissible concentration of ammonia in
atmospheric air has not been established thus far. The object of our study
was to give an experimental validation of the highest single and mean daily
maximum permissible concentrations of ammonia in atmospheric air.
The gas in the experiment was obtained by using a 25% solution of
ammonia in a distilling flask.
The concentrations of ammonia in air were determined by a colorimetric
method based on the yellowish-brown coloration of solutions formed when
ammonia acts with Nessler's reagent. The sensitivity of the method is 0.3yg
of ammonia in the volume analyzed.
- 104 -
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To validate the highest single maximum permissible concentration of
ammonia in atmospheric air, we determined the threshold of olfactory per-
ception by using a method proposed by the Committee on Sanitary Protection
of Atmospheric Air (1957). The tests were conducted on 22 practically
healthy persons aged 17 to 48 years. A total of 432 tests with concentra-
tions from 5 to 0.4 mg/m^ were performed (Table 2).
Table 2
Threshold of Olfactory Perception of \rjnonia.
Number of
Subjects
Number of
Observations
Concentration _cf_NH_j,_jng/mJ
Minimum I Kaxinun
3ercep-„ible Imperceptible
36
138
200
58
1,95
0,98
0,70
0,50
I
1.0
0,75
0,55
0,45
As can be seen from Table 2, the threshold of olfactory perception of
ammonia in the most sensitive persons is 0.5 mg/m^, and the subthreshold
concentration is 0.45 mg/m^.
Numerous studies established that fragrant substances in concentrations
of undetectable odor can cause definite reflex responses in the human organism.
Such reflex changes arise in the visual system and in the cerebral cortex,
and we utilized them in the study of the subsensible effect of ammonia on
the human organism.
Adaptometric studies were performed with the aid of an ADM adaptometer
on 3 subjects whose threshold of olfactory perception of ammonia was first
determined.
The tests were conducted with concentrations of 0.65 and 0.51 mg/m^
for one subject and 0.51, 0.45, and 0.32 mg/m^ for the other two. This
distribution was due to the subjects' different thresholds of perception of
ammonia odor. The effect of each concentration was studied no fewer than
three times. The data obtained show that ammonia in a concentration of 0.45
mg/m^ decreases the light sensitivity of the eyes in two subjects (D. N. and
L, N.), whereas a concentration of 0.32 mg/m^ was found to be inactive for
them (Fig. 1). In the third subject (Sh. L.), the threshold concentration
for the change of the light sensitivity of the eyes was found to be 0.65
mg/m^, and the subthreshold concentration, 0.5 mg/m^.
To determine the threshold of the electric activity of the brain, we
used the method of recording the flare-up of the alpha rhythm during the
action of intermittent light timed to the intrinsic potentials of the brain.
- 105 -
-------
The work was performed in the department of communal hygiene of the Central
Institute for Advanced Training of Physicians.
*0 000
30 000-
f///
10 000
5 to >5 20 25 JO 35 ii>
Minutes
Fig. 1, Change in the light sensitivity of the eye
in subject D. N. during inhalation of different
concentrations of anxonia.
1 - pure air; 2 - concentration 0.32 ag/ni*; 3 - con-
centration 0.45 mg/n;3; 4 - concentration 0.5 mg/=-'.
The studies were conducted on 5 subjects aged 18 to 24 years, whose
ammonia odor threshold was first determined. The changes in biopotentials
were recorded on an 8-channel Kaiser electroencephalograph. The total
activity of the human cerebral cortex was analyzed with the aid of a multi-
channel integrator of B. N. Balashev's system (1964). The biocurrents were
taken off in the bipolar manner. Rhythmic photic stimulation was carried
out by means of a photostimulator at a frequency of 8 flickers per second
and an intensity of 0.1, 0.2, and 0.6 J. Three observations were carried
out with each concentration, and 2-3 observations with pure air for the
control. The total bioelectric activity for the entire period of action
of the light was calculated by means of an integrator. . The results ob-
tained were expressed in percent. The average acticity calculated for the
first three minutes was taken as 100%.
In 3 subjects with an odor threshold of 0.55 mg/m^, a concentration of
0.35 mg/m^ had a substantial effect on the electrical activity of the brain;
in two subjects (Kh. V. and K. V.), an attenuation of the total bioelectric
activity was observed during the first minute of supply of the gas, and a
reinforcement was observed in subject D. M. In all three, an ammonia concen-
tration of 0.22 mg/m^ caused no changes (Fig. 2).
In two subjects (D. N. and I. N.) with an odor threshold of 0.76 mg/m^,
the active concentration was found to be 0.44 mg/m^, and the inactive concen-
tration, 0.32 mg/m^. Ammonia had the most pronounced effect in the fourth
-106 -
-------
minute of the experiment with a gradual attenuation in the 5th-6th minute.
A more pronounced effect on the organism when ammonia first begins to act
is also observed when the threshold of olfactory perception is established
and when the light sensitivity of the eyes is studied.
03
¦V
=
¦P c
0)
E o.
5 e 7 8 9 10 II
Mir.utes
Fig. 2. Amplitude of breir. potentials in subject K. V.
during inhalation cf different concentrations of srjronia.
1 - pure air: 2 - concentration 0.22 ag/ni?; 3 - concentra-
tion 0.35 «g/m3. Arrows indicate the time of inhalation
of the &as.
Table 3 shows the thresholds of the reflex effect of ammonia on man.
Thus, the concentration of 0.2 mg/mwhich was found to be the sub-
threshold value according to the most sensitive method, may be recommended
as the highest single maximum permissible concentration for atmospheric air.
Table 5
Thresholds of the Reflex Effect of Arrnonif..
Effect Studied
Concentration, ms/m5
Threshold ^Subthreshold
Olfactory perception
Change of the light sensi-
tivity of the eyes
Change of the electrical
activity of the brain
0.50
0,45
0,45
0.32
0,35
0,22
In order to establish the mean daily maximum permissible concentration
of ammonia in atmospheric air, we studied its chronic effect on the animal
organism. A continuous round-the -clock exposure of white rats to gaseous
ammonia was carried out for 84 days in the following concentrations: group
I, 20 ± 0.1 mg/m^ (level of the maximum permissible concentration for indus
trial buildings), group II, 2.0 ± 0.061 mg/m^, and group III, 0.2 ± 0.0072
mg/m^ (level which we propose for the highest single maximum permissible
concentration for atmospheric air); group IV was the control.
- 107 -
-------
Sixty white male rata weighing 105-150 g were selected for the chronic
exposure. The latter was carried out in 100-1 chambers. Purified air mixed
with gaseous ammonia in definite concentrations was supplied to the chambers
at a rate of 28-30 1/min. Samples of air for analysis were taken from the
chambers every day.
In the course of the chronic experiment, the general condition of the
animals, their weight, state of the latent reflex time, porphyrin metabolism
and ammonia content In the urine were observed. In the blood of the animals,
the cholinesterase activity, oxidation-reduction function, number of fluo-
rescent* leucocytes, hemoglobin, erythrocytes, and nucleic acids were deter-
mined. At the end of the exposure and recovery period, part of the animals
were killed and subjected to anatomico-pathologic analyses. The results of
the experiment were evaluated by calculating the reliability between the
indices of the control and experimental groups of animals, obtained by statis-
tical treatment using the range method.
In the course of the experiment, the rats were healthy and active in all
the groups, and no weight lag was observed in comparison with the controls.
Measurement of the latent tine of the reflex response permits an evaluation
of the functional state of the central nervous system for different actions
of toxic substances on the organism (A. A. Minayev, 1965; A. P. Fomin, 1965).
We used for this purpose a "chronoreflexogenometer" instrument proposed by
S. I. Gorshkov (1964). The time (in sigmas) of the appearance of the motor
response to the action of the pain stimulus (electric current) was determined.
Results of the studies are given in Table 4 and Fig. 3
In our studies, a significant shortening of the reflex time was observed
during the first month of. exposure only in group I during the action of ammonia
in a concentration of 20 mg/m^.
We determined the cholinesterase activity of the blood by the Fleisc'ner-
Pauns procedure modified by N. K. Pushkina and N. V. Klimkina (N. N. Pushkina,
1963). A depression of cholinesterase activity occurred in rats of group I.
No changes were observed in the other groups (Fig. 4).
The oxidation-reduction function of the blood serum was determined by
using a modified method of Tumberg (Yu. L. Anin, 1964). The method is based
on the change in the color of methylene blue with blood serum during boiling
on a water bath and the determination of the time of its total bleaching.
* Editor's r.ote: "or the Russian use of the terns "luminescent" and "luminescence" ir. this p'.per, we have
substituted "f.'j orescent" and "fluorescence", an the basis of the definitions of these terms.
- 108 -
-------
A lengthening of the time of total bleaching as compared with the control
group occurred in rata of groups I and II (Table 5, Fig. 5). This phenomenon
characterizes a depression of the oxidation-reduction processes in the animal
organism.
Sigma s
80
70
i
E*
S.
SO
so
-
/
1 s
J
\ 2
\
_
V
/
1 '
1
Periods of study
?ig. 3. Change in the latent reflex time in rats under
the influence of different concentrations of ammonia.
1.- group I (20 og/m'); 2 - group II (2 ir»j/ra-/);
5 - group III (0.2 - control group IV;
AE - period of exposure
Table ^
Charjge in th? latent Reflex Time injtats
Group
/
Periods
<20
" T
(2 me/r.-)
111 7
(0.2 -g/m')
IV
(control
Before exposure
1st rr.cnth exposure
2r.d month exposure
3rd month exposure
Recovery
period
71,4(o)
57.6(c)
66,3(0)
68.3(o)
68,0(o)
69,2(o)
66,4(o)
68,0(o)
68,4(o)
68.8(c)
70,4(o)
67.8(a)
67 ,C(o)
68.4(o)
68,0(o)
71.3'
67.0
68,3
68.1
68.2
«ote. Degree of significance:
99.9%; o - insignificant
One of the methods of determining the early qualitative changes in the
formed elements of white blood cells is fluorescent microscopy. The method
is based on the ability of nuclear nucleoproteins of degenerating cells to
combine with acridine dyes in a different manner than the nucleoproteins
of undamaged cells (M. K. Meysei' and V. A. Sondak, 1956). However, during
the exposure we failed to observe any degenerative changes in the leucocytes
in the blood of the experimental rats as compared with the control group.
- 109 -
-------
Nor were we able to detect any changes by studying the blood for the content
of erythrocytes, hemoglobin and total quantity of nucleic acids.
m
m
2i0
tyxu 20/XH 30/Xu 23!< 15/H Sim jo/in tl/iy 25! 5/'
Dates of study
Fig, 4. Change of cholinesterase activity in the blood
of rats under the influence of ammonia.
Notation same as in Fig. 3.
Table 5
Change of the Oxidatior.-Reduction Function of the Blood Serar.
in Rats (.Ti=e in Minutes)
G_rou£
Periods
(20 mg/n5)
H ,
(2 Eg/m5)
III , :v
10.2 mg/m5) (Control)
Before exposure
Exposure
Recovery period
period
I0,53(o)
19,10(c)
12,66(o)
10,93(o)
15.0(c)
12,73(0)
I0,4€(o) 1 10,66
12,20(0) 12,05
12,63(o) 12.66
Note. Confidence factor: c - '99.9%; o - insignificant.
In order to characterize the chronic action of ammonia on the organism
of the experimental animals, we also studied the porphyrin metabolism.
Yu. K. Smirnov (1953) showed that excitation of the nervous system
causes an increase in the porphyrin metabolism, and inhibition causes a
decrease. M. I. Gusev (1960) was the first to study the porphyrin metabolism
in connection with a sanitary standardization of lead in atmospheric air.
Subsequent studies by G. I. Solomin (1961), Li Sheng (1961), V. I. Filatova
- 110 -
-------
(1961), B. M. Mukhitov (1963), and others confirmed the high sensitivity
of this method. The content of coproporphyrins in the urine was determined
spectrophotometrically. It was found that during the exposure, an increase
in coproporphyrin occurred only in group I of the animals (Fig. 6).
fi B
.
/ '
¦ • •.
-
1
"A
» *
i V
f
1
u
ji
/
*V
- J Wll
\
¦ _ K
1 . . 1
1 » < t
1 1 I
U/JU1 20/XH 30/M ,V//. 15/11 s/m JD/m 12Itv ZS/IV S/v
Dates of study
Fig, 5. Kffect of ammonia on the oxid3tion-reriuction
function cf the blood serum of rats.
Notation same as in Fig. 3.
Simultaneously, we determined the content of ammonia in the urine of
the animals. An increase in the ammonia content was also noted only in
group I.
From the round-the-clock chronic exposure of white rats which was per-
formed, it was found that ammonia in a concentration of 20 mg/m^ caused a
shortening of the time of the reflex response,- a depression of cholinesterase
activity and oxidation-reduction function of the blood, and also an increase
in the excretion of coproporphyrin and ammonia in the urine.
Ammonia in a concentration of 2 mg/m^ caused only a depression of the
oxidation-reduction function of the blood serum. A concentration of 0.2 mg/ni^
was found to be inactive.
Histopathological examinations did not show any changes in the internal
organs and central nervous system in animals of the experimental groups as
compared to the control group.
- Ill -
-------
Mg per iOC g of weight
aso
aso
ato
Dates of study
Fig, 6. Effect of ammonia on the coproporphyrin exchange of rats.
Notation same as in Fig. 5.
Conclusions
1. A whole series of enterprises including metallurgical plants whose
composition includes coking and nitrogen fertilizer sections constitute major
sources of pollution of atmospheric air with ammonia.
2. The subliminal concentration of ammonia which did not cause a change
in the biopotentials of the brain, equal to 0.2 mg/m^, is proposed as the
highest single maximum permissible concentration in atmospheric air.
3. A similar concentration of ammonia (0.2 mg/m^) during around-the-
clock chronic exposure was found to be inactive and can therefore also be
recommended as the mean daily maximum permissible, concentration in atmos-
pheric air.
LITERATURE CITED
Note: References mentioned in this paper are to be found at the end
of the volume in the 196 7 bibliography.
- 112 -
-------
POLLUTION OF ATMOSPHERIC AIR WITH VAPORS OF HYDROLYTIC ETHYL ALCOHOL AND ITS
EFFECT ON THE ORGANISM
Candidate of Medical Sciences R. Ubavduliayev
A, N, Sysin Institute of General end Communal Hygiene of theUSSR 4c»de=y of Medical Sciences and
Izfcek Scientific Research Institute of Hygiene, Sanitation, and Occupational Diseases
From Akaderaiya Heditsinskikh Nauk SSSR. "Biologicheskoe deystvie i
gigienicheskoe znachenie atmosfernykh zagryazneniy". Red. V, A. Ryazanova.
Vypusk 10, Izdatel'stvo "Meditsina" Moskva)/:p. 74-86„ (196?)
Hydrolytic alcohol is ethyl alcohol (ethanol) containing different impuri-
ties of acids (in terras of acetic acid, 0.036-0.12 g/1), unsaturated compounds
(in terms of allyl alcohol, 0.07-0.89 g/1), 3-6.2 g/1 of methyl alcohol, car-
bonyl compounds (in terms of acetaldehyde 1.7-21.6 g/1), 0-0.24 g/1 of furfural,
and higher alcohols (in terms of isobutyl alcohol, 2.3 g/1).
Ethanol is a transparent, colorless, volatile liquid with a specific odor
and a boiling point of 78.4°C. Ethanol is miscible in all proportions with
water, ether, and chloroform; it dissolves inorganic salts, particularly
chlorides, nitrates, and acetates, essential oils, and some fatty oils.
Ordinary ethanol is obtained by alcoholic fermentation of such starch-
containing materials as potatoes, cereals, rice, and also molasses; synthe-
tic ethanol is obtained from ethylene and acetylene.
In the ethanol-producing industry hydrolytic alcohol has become the most
common product. Its production is based on the fermentation of hexose sugar,
obtainable from hexose-containing plants. Ethanol finds applications as a
solvent in the manufacture of lacquers and varnishes, and is used as the
starting material in the synthesis of many organic compounds, in the prepara-
tion of synthetic rubber by Lebedev's method, as a fuel for internal combustion
engines, in the food industry, in medicine, and in many other branches of the
national economy.
In the nature of its action, ethanol is a narcotic and a nerve poison.
In high concentrations, it first causes stimulation, and then paralysis of the
central nervous system. Prolonged chronic exposure to large doses may cause
serious organic diseases of the nervous system, digestive tract, cardiovascu-
lar system, liver, etc.
There are no literature data on the effect of low ethanol concentrations
on man and animals during inhalation. Its maximum permissible concentration
in atmospheric air has not been established. Nor are there any data on the
pollution of atmospheric air with ethanol around the plants producing it. The
object of the present study was to elucidate these questions.
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The determination of ethyl alcohol in atmospheric air was made with the
aid of I. A. Pinigina's method (1961), based on the reaction of alcohol with
the vanadium-hydroxyquinoline complex with the formation of an orange color.
The sensitivity of the method is 0.002 mg in a volume of 2.5 ml. To obtain
the desired concentration in the experimental mixture of air and ethanol,
we passed air through a distilling flask filled with hydrolytic alcohol.
Before the start of the experiments, the constancy of the ethanol concentra-
tions in the cylinder was studied for several days, and found to remain at the
same level with only slight fluctuations. To determine the odor threshold,
25 persons aged from 18 to '40 years were chosen. A total of 385 determinations
of the odor threshold were made with 7 concentrations (from 14.8 to 6.3 mg/m3)
(Table 1).
Table 1
Results of Determination of the Odor Threshold
of Hydrolytic Ethyl Alcohol
Number of
Subjects
Cor.centration, m/m*
Minimum
Perceptible
Maximum_
Imperceptible
i
10.2
9,4
7
8,7
8.0
7
8,0
7.3
4 ,
7.3
7,0
6
7,1
6.3
Thus, the threshold of olfactory perception of hydrolytic alcohol in the
most sensitive persons of this group was found to be 7.1 mg/m^, and the sub-
threshold concentration, 6.3 mg/m^. We then determined the effect of low
ethanol concentrations on the light sensitivity of the eye by the dark adapta-
tion method. A total of 42 tests on persons aged 18 to 27 years were carried
out. Concentrations of 8.29, 6.97, and 6.12 mg/m^ were studied.
An ethanol concentration of 8.29 mg/m^ caused a change in the course of
the dark adaptation curve in all four subjects. For the most sensitive two
person, the minimum active concentration of ethanol with respect to the light
sensitivity of the eyes was found to be 6.97 mg/m^. The inactive concentration
for these persons was 6.12 mg/nP (Table 2).
In studying the effect of low ethanol concentrations on the electrical
activity of the cerebral cortex, we used A. D. Semenenko's method (1963) of
a reflex response involving a flare-up of the alpha rhythm in man during simul-
taneous stimulation of the subject with intermittent light whose frequency
corresponds to his rhythm and with sound whose intensity was varied continually
Superimposed on the background of the functional load was a reinforcement of
the intrinsic alpha rhythm of the subject, and under the influence of the inhaled
gas mixture with the active ethanol concentration, there was a change in the
character of the recorded surves, indicating a change in the dynamics of the
nervous processes occuring in the cortex of the cerebral hemispheres.
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Tatle 2
Threshold of Reflex Variation of the l.ight Sensitivity of the Ejre
Duri Inhalation of Vapors of !lydrolyt\c Ethyl Alcohol (in 20th
minute in percent of 15th -in'-it*.;
Subject
Pure Air
8,29
Coneentra
6.97
tion. mg/r
u.12
Z
~
Sut-
thres-
t.old
I. L.
3. Ye.
Ch. A.
E. V.
158,6 (o)
156,0 (o)
133,5 (o)
148,0 (o)
194.8(b)
226.7(b)
223,6 (c)
214.4(b)
150,0 (o)
155.6 (o)
151,9 (b)
130.7 (b)
— 1 8,29
— j 8,29
134,3 (o), 6,97
147,8 (o) 6,97
6,97
6,97
6,12
6,12
Note. Degree of significance: e - 95&; b - 99%; s - 99.
o - insignificant.
The curves were recorded on an eight-channel Hungarian electroencephalo-
graph of the Orion Budapest Co. The study was made on five persons with the
lowest odor threshold under the same conditions and at the same time.
Ethanol concentrations of 6.14 and 4.9 mg/m^ were studied. The experiment
was conducted no fewer than 4-5. times with each concentration. Results of the
study and analysis of the statistical treatment of the data show that a concen-
tration of 6.1 mg/m3 was active for all five subjects. The inactive concentra-
tion was 4.9 mg/m (Table 3).
Results of all the studies of the reflex effect of ethanol are summarized
in Table 4,
On the basis of the study of the reflex response of man, we propose 5 mg/m3
as the highest single maximum permissible concentration of hydrolytic ethanol
in atmospheric air.
To validate the mean daily maximum permissible concentration of ethanol
in atmospheric air, we conducted a 90-day round-the-clock dynamic exposure of
45 white male rats weighing 100-120 g, which were divided into three groups.
In selecting the concentrations for the exposure, we used the results of
the study of the reflex effect of ethanol on the human organism. Group II of
rats was exposed to ethanol vapors in a concentration of 5 mg/m^ at the level
of our proposed highest single maximum permissible concentration, and in group I,
the ethanol concentration was five tiroes as high.
Group III of the rats was the control group. To evaluate the effect of
ethanol vapors on the organism of the animals, we studied the behavior and
weight dynamics of the animals, motor chronaxy of antagonist muscles, excre-
tion of coproporphyrins with the urine, activity of blood cholinesterase, to-
- 115 -
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tal proteins, and protein fractions of the blood serum. All the numerical data
obtained were subjected to statistical treatment.
Table 3
Effect of Low Concentrations of Hydrolytic Etnanol on
the Electrical Activity of the Cerebral Cortex
Subject
6,1 ag/W
Hemisphere
4.9 mg/m3
Hemisphere
Left
Right
Left
Eight
I. s.
~
L. U.
-r
+
—
S. K.
+
—
—
L. B.
4-
"t-
—
U. K.
+
~r
—-
—
Mote: + statistically significant changes,
- insignificant changes.
Table <*
Effect of Low Concentrations of Hydrolytic Ethanol
on Receptors of Respiratory Organs.
Function
oncentration, ms/m*
Threshold. Subthres
Odor perception
7.1
6,3
Light sensitivity of eye
6,9
6,1
Slectrical activity of
4,9
cerebral cortex
6,1
The experimental mixture of air with a given ethanol concentration was
supplied to the chamber at the rate of 35 1/min. Such a rate of supply of air
produces the most favorable conditions for the animals (V. A. Popov, 1964).
The actual ethanol concentrations in the chambers were: in group I, 29.25 -
2.1 mg/in^, and in group II, 5.59 - 0.45 mg/m . During the exposure, the animals
of all the groups were healthy, active, and gained weight moderately. However,
a slight weight lag was observed in rats of group I. We determined the chrona-
xy of antagonist muscles every 10 days in five rats of each group at the same
time and under the same conditions, using an ISE-01 electronic pulse stimulator
(Table 5 and Fig. 1).
The above data indicate that in rats of group I, a distrubance of the normal
ratio of the chronaxy of extensors and flexors occurred on the 6th day of exposure.
At the end of the recovery period, all the ratios returned to normal.
In rats of group II, there were no statistically significant changes as
compared with the animals of the control group. The activity of whole blood
cholinesterase was determined by the method of A. A. Pokrovskiy (1953) and
A. P. Martynova (1957) in five rats of each group.
- 116 -
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y\
w
Dates of study
Fig. 1, Change ir. the ratio of ohronaxy of antagonist muscles
during inheletion of vapors of hydrolyiic ithanol.
A-E - period of exposure; 1 - pure airj 2 - concentration
29.25 ng/i5; 3 - concentration 5.5? og/m}.
Taole 5
Ratio of Cfcronaxy of Antagonist Muscles in Rats During Ir.halR-
ticn of Vapors cf Hytfrolytie Sthanol
Periods
Date of
Study
Greun
1
11 • 1 Control, III
Ratio of Chronaxia of Extensors to
Chronaxy of Flrxcrs
Before exposure
23/IV
1,62 (0)
1,53 (0) j
1,65
3/V
! ,29 (0)
1,33 (0) .
1.45
13/V
1,30 (0)
1,43 fo) j
1,58
Exposure
22/V
1,55(0)
1,50 (o)
1 . o4
31/V
1,26 (0)
1,58 (0) |
1,48
9/VI ,
1,37 (0)
1,53 (0) 1
1 ,33
19/VI
0,82 (c)
1,42 (o) '
1 ,41
29/VI
0,93 (c)
1,49 (o) ;
1,54
, 9/VH
0,91 (b)
1,35 (0}
1,57
18/VII
1,1 (0)
1,42 (o) !
i, 14
29/V! 1
0,91 fa)
I ,26 to) j
1 , 14
i 1/VJll
0,79 (0)
1 ,56 (0) 1
1,39
Recovery
23/VIH
1,08 (0)
U5 (o) [
1,22
L/1X
1,23 (0)
1,09 (0) !
1,23
N;te. Degree of significance: a - 9%5 0 - 99.9%; c - 99.9$;
0 - insignificant.
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Table 6
Change of Cholinesterase Activity in Rats During Inhalation of
Vapors of Hydralytic Ethanol (in minutes of 1-ydrolysis of
acetylcholine).
Period
Date
(29.?
Groups
1! ,
ng/n5)
Control
ill •
Before exposure
28/IV
37,6 (o)
3?,0 (o)
39.2
£
13/V
37,8 (o)
38,8 (o)
39,2
X
24/V
39,8 (o)
31,4 (o)
38,0
7/V1
39,0 (o)
39,2 (o)
38,0
0
21/Vl
42.0 (c)
38,6 (o)
38.2
s
5/Vll
-12,2 (b)
33,8 (o)
38,8
u
19/VII
39,8 (o)
37,4 (o)
38, S
r
2/Vlli
41,8 (b)
37,8 (o)
38,0
e
12/Vill
¦12,2 (a)
37,8 (o)
37,6
Recovery
I/IX
38,7 (o)
38,0 (o)
37,4
Note. Degree of significance: ¦: - 95$; - 99^>; c - 99.9&;
o - insignificant.
Minutes
Exposure
¦ 1 I I 1 I I LJ—
2t.In i3b tin
-------
ani-nal organism. After 20 days of the recovery period, the time of acetylcho-
line hydrolysis returned to normal. No changes were observed in rats of
group II as compared to the control.
nig
o as
27/W i/V '//V 2S/V S/Y! tt/vt 6/vn I5im. 3/HH :S/HU ?S/W> 5/:/
Dates of study
Fig. 5. Change in the excretion of coproporphyria with the
urine during inhalation of vapors of hydrolytic ethanol.
Notation same ,as in Fig. 1.
We evaluated the effect of low concentrations of ethanol vapors on the
metabolism from the change in the porphyrin metabolism in the animal organism.
The coproporphyrin was determined by Fischer's method. The extraction of
porphyrin from the urine was carried out with ether, and the quantitative
determination, on an SF-4 spectrophotometer in the 400-410my region. The
daily portion of urine from each group of animals (with 5 rats in each group)
was collected in special glass containers.
Coproporphyrin was determined once every two weeks, and its amount was
recalculated in terms of 100 g of the animals' weight. In rats of group I,
the excretion of coproporphyrin with the urine starting with the 6th week
of exposure decreased sharply, and remained at this level until the end of
exposure (Fig. 3), Normalization occurred at the end of the recovery period.
No statistically significant changes were found in rats of group II as
compared with the control group. In the chronic experiment, the effect of
low concentrations of ethanol vapors on the total proteins was also studied,
and their fractional composition in the blood serum was determined with the
aid of paper electrophoresis. The blood was taken on an empty stomach, from
the tail every 15 days. In rats of group I, by the end of the 2nd month of
exposure, the content of albumins decreased, and the gamma globulin fraction
increased. This increase was statistically significant and lasted until the
end of exposure. Even during the recovery period, the albumin-globulin ratio
did not return to normal (Table 7 and Fig. 4),
- 119 -
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'00 -
/3/y 26/* to/vi 23/V! 7/rn P/"U t/m 13/vm i/>i
Dates of study
Fig, k. Change in the protein fractions of the blood serum
during inhalation of hydrolytic ^thanol vapors.
Notation sane as in Fig. 1.
No changes whatsoever were observed in rats of group II. No
change was noted in the total proteins of the rats in any of the groups.
Thus, continuous exposure for 90 days had a marked effect on animals of
group I (29.1 * 2.1 mg/nr), but had no effect on those of group II (5.59 -
0.045 mg/nr*). On the basis of these data, we can recommend that the mean
daily maximum permissible concentration of hydrolytic alcohol be taken at
the level of the highest single concentration, 5 mg/m3. In order to obtain
a sanitary evaluation of atmospheric pollution with vapors of hydrolytic
ethanol, we conducted a survey of three hydrolysis plants in Uzbekistan. A
total of 186 air samples were collected at distances of 100, 200, and 300
meters on the leeward side of the source of discharge. The samples were
collected by suction into two absorbers containing activated carbon. From
10 to 40 1 of air was collected in each sample at a rate of 1 1/min. The
air temperature during the sampling ranged from 20 to 353C, and the relative
humidity, from 25 to 70%; the barometric pressure was 730-750 mm Hg, and the
wind velocity, from 1 to 5 m/sec. During the sampling, the weather was dry
and clear (Table 8),
It is evident from Table 8 that at a distance of 100 m and farther from
the source of discharge around the three Uzbekistan hydrolysis plants studied,
the ethanol concentrations were below our proposed highest maximum permissible
concentration of 5 mg/m .
- 120 -
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Table ?
Change in the Total Proteins and Protein Fractions of the Blood
Serum During Chroni~ Exposure of White Hets to Vapors of Hyaro-
lytic Rtt-inol
Period
Group
Total
Proteins
Albumins
21
Alpha
obulins
1
Betta jGarrusa
Albuein-
Globulir.
Ratio
Before exposbre
I
II
HI
(Control
,9,09
9,05
8.91
)
42.3
42,6
45,1
23.8
22.9
22,9
19,5
20,4
19,4
14,4
14,1
12,6
0,73
0,74
0.83
On 50th day
of exposure
1
I 1 8,89
II j 9,04
HI , 9,88
(Control^
|
32,3 1 24,6
42.6 | 21,6
43.7 , 20,8
26,0
21,4
20,9
17,1 0,48
1 1.1 : 0.71
14,6 0,77
j
On 90th day
of exposure
I
II
III
(Control.
9,40
8,63
8,73
i
35.3 | 23,4
45.4 { 21,1
42,9 | 21,8
23,5
21,1
21,!
16,8
12,4
14,2
0,54
0,84
0,76
Recovery
X
9,38
8,67
8,79
37,9
44,8
44,2
24.0
21.1
19,8
22,6
21,8
23,4
15.5
12,3
12.6
0,60
0,83
0,80
Hesults of Study of the Pollution of Atmospheric Air with Sthanol
Vapors Around Hydrolysis Plants in Uzbekistan
Distance
Pros
Discharge
m
Number
of
Samples
Single Concen-
trations, ng/m3
N-j-ber of Samples
centratior. of,
with Con-
mg/r.3
V.axizum
Minimum
2 ar.d
above
Frc~
1 to 2
Beiow '¦
Fergana Plant
100
26
2,79
0,55
6
9
1 11
200
17
0,16
! 1"
300
27
0,018
—
—
27
Andizhan Plant
100
25
4,52
0,45
1 10
10
: 5
200
16
1,68
—
1 2
! !•'<
300
27
0,37
—
i __
i ¦ —
1 26
Yangiyul' Plant
100
44
2,2
4
10
10
200
25
0,5 1
—
2">
- 121 -
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Conclus ions
1. The atmospheric air around the surveyed Uzbekistan hydrolysis plants
is slightly polluted with vapors of hydrolytic ethanol.
2. A study of the biological effect of hydrolytic ethanol vapors showed
that the threshold of olfactory perception in the most sensitive persons is
7.1 mg/m^. The threshold of change of the light sensitivity of the eyes is
6.97 rag/m3, and the threshold of the reflex effect on the activity of the cere-
bral cortex is 6.1 rag/m3. The maximum inactive concentration according to the
most sensitive test is 4.9 mg/m .
3. The highest single maximum permissible concentration of hydrolytic
ethanol may be established at a level of 5 mg/m3.
4. Chronic round-the-clock exposure of the experimental rats to ethanol
vapors in a concentration of 29.25 mg/m3 for 90 days caused changes in the
normal ratio of the chronaxy of flexors and extensors, cholinesterase activity,
excretion of coproporphyrin with the urine, and in the relative amounts of the
protein fractions of the blood serum. Ethanol in a concentration of 5.59 mg/
had no effect on the rat organism.
5. The mean daily permissible concentration of hydrolytic ethanol in
atmospheric air based on the data of the chronic experiment may be recommended
at the level of the highest single concentration, 5 mg/m3.
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Note: References mentioned in this paper are to be found at the end of
the volume in the 196 7 bibliography.
- 122 -
-------
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• AySpoBCKan . H. B kii : ripcAc.ibtio jonycTinibie Koimcinpaanu
aTM0C(}tepHUx 3arpn3iicHiifi. B. 3. A\.. 1957, CTp. 44—62.
£ w *: e b a IO. B. B kh.: yncMbie aanucKii Mockobckoto nny. . Spiic.Maiia B. 5.
At., i960.
E ji $ ii m o b a E., fl y tii k ii ii a H H. Tur. » can., I9GG, 2, 85—86.
)KH/i0 8a H. A. Hir. n can., 1959, 12. 18—23.
3 a k y t ti h c k it ft JX. 11., n a p $ e n o b IO. J1, C e a ii b a it o o a /I. H.
CnpasoMMifK no TOKCiiKOAoriiu paanonKTnoMux liaoTonoc. Me.ani3,
1962.
- 124 -
-------
3apoilo b c k h ft Tl. 0. BecTH. AMH CCCP, 19G4, 9, 8G—91.
3 a p o a o a c k if i"i n. . Bccth. AMH CCCP, 1901, 4, 9—19.
H fi p a r ii m o b M. T., Jl n c h h ii c k ii i'i E. 3. B kit.: Tpyau oTO.ia-
piiuro/ioroa BauKitpcKoft ACCP. B. 3. y$a. 1902, crp. 16—22.
H M a uj e b a H. B. Tur. i? caH., 19G3, 2, 3—8.
H ui h mom Jl. M. B kh.: XV HayMiian cccciih MeanumicKoro iihcth-
TyTa. Te3HCbi aok/isaob. Kyi*i6uujeB, 1951, 57—58.
H u k o b ii 4 A. A;, B ii h o r p a a o b a B. A. B kii.: Teaiicw AOK/iaaos
, ' HayiHOfi ceccc.ii HoBOCHfiiipCKoro camiTapiioro iincTiiyyTa. Hobocii-
CHpcK, 1956, crp. 23—26.
K a fo >k h w ii f\. H., T o p a kf h s M. n. B km.: MaTep:iaapMaKo.norntt aueTo;})CKOHa.
&HCC. AOKT. cn B. 1878.
K a n fl'a h c k ii ft C. fl. BecTH. AMH CCCP, 1962, 9, 10—21.
KHa6eiiro$ B. I\ Ttir. Tpyaa.H Tex. 6c3on., 1934, 4, 22—26.
KopOaKoea A. H., KpeMHCBa C. H., Ky^iariiHa H. K.,
- y A a H o b a M. n. B kii.: Ilpo.Mbiiii^eiiHaH tokchko-iohih. M., I960,
crp. 232—238.
KopxeeB 10. E. Tiir. h can., 1965, 9, 15.
KouitorhuX. C. lOCjMefiHhifi cfiopniiK AH CCCP. M., 1947,437—464.
Kpa skob C. B. r^a3 ii ero pa6oTa. M.—Jl„ 1950.
KynpHU E H. Tur. Tpyaa h npo$. 3a6o.ieBaHiin, 1925. I, 80—81.
Jlaspoe E. A., ft h o b e k a a B. M. B kii.: BirraMHiiu. T. 2. Knen,
1956, crp. 61—64.
Jl as apes H. B. B kh.: Bpeairue BetnecTBa b npOMUtiweHitocTit.
M. I. rocxiiMH3/iaT. J\., 1951, ctp lJ2— I3l, 323.
Jlasapes H. B. B kii.: BpeA'iwc BcmccTaa b npoMuui/iemiocTH.
4. I—II. Jl., 1963.
Jl H IIIsh. rnr. a can , 1961. 8, 11.
/lioOHMoaa M n. CfiopniiK 3KcnepHMc(iTa.it>'ibix Hcc/ieAOBaiiiifi no
npoMUUJfleiiHtiN waM. B. XXV. .fl., 193G. CTp. 29—36-
MaHiiaHOoa X. X. B kii.: npeAc.ibno AonycTiiMHc KonuciiTpauHM
aTMocc{iepi:w.\ 3arpn3iieintii. B. 8; M, 190!, CTp. No—160.
MepKos A. M. 06tnan Teopna h meTOAHKa caiiHTapuo ctaTHCTHie-
CKOro licc.ieAOBainin. M.. 1900
Mhji/icp C. B. B kii.: npcAe.tbiio AonycTitMMe KoimciiTpaumi amo-
ca, 1964, CTp. 4—20.
MHxe^bcOH M. S. jXet'icTBiis iiapKoniKOD na xo/imiscTepasy. Jl.,
1948.
MyxiitoB 5. M. B kii.: npeae/tbiio AonycTiiMbie KOHucnipamm aT«o
• cepn'bix 3arpH3iiciiiit"i. B. 7. M., 19G3, cip. 76—98.
Mutiihk n., reHKii.tr C. B kh,: Tpyaw XI cte3Aa TcpaueoTos
CCCP. rocMCAHia, 1932, CTp. 253.
H a 3 u p o u T, H. McAimiiiiCKim >k>pna/t yjCcKiicrana, 1960, 8, 28—32.
Heft Ma ii KD. B„ Cmoahii B. B. B kii. Tpyau Bcecot03noro cone-
LuaiinH no aounite ct nw-nii ii ra30D. M.. 19G1
Hobkkob 10. B. B kii.: IlpcAc/ii.iio aonycTiiMbic KoituciiTpaUHii atMo-
c4>epnu.K 3arpfl3iiCHin"i. B. 3. M., 1957, ctp. 85- 107.
- 125 -
-------
0 a t> ui a h c k h ft M. H., /I h x a m e b a B. B. Tpyaw PocroncKoro'iia-
Aofiy MeAiimiiiCKoro HiicittTyTa. C6. 2, PoctoB iia-Jloiiy, 1935,
crp. 120.
IlflBJioB H. n. /Ickuiih no H3Hin. H32. AH CCCP, 7!.—M, 1952.
O n a p h ii A. H„ E d p c mi o b a T. .H. Bcctii. AH CCCP, 1917, 5®. io3.
IletpOB A. M. B kii.: Tpyau /IcHimrpaiicKoro caii.-rnr. mcx hiictk-
TyTa. A., I960, CTp. 153.
norocHH y. T. Fiir. h can., 1965, 7, 3.
F1 o k p o b c k ii ft A. A. Boen. uca. xcypit., 1953, 9. 61—65.
II peo6 p a )K e k c k ii i'i B. C. BecroAMH CCCP, 1964, 10, 7—16.
flyiiiKHHa H. H. BuoxiiMtwcKHe MeToau ucaie/waaium. M., Mca-
IW3, 1963.
ribRHKOs B. B. JKypii. oCiuch xhmiui, 1933, 3, 6.
nuuKiiii B, H. Becnt. AMH CCCP. I9G3. 4, 20-28.
PcBKosa H. B. rnr. tpyfla h npo4>. 3a6o.ieBanitfi, 1955, 2, 17—2?.
PaJCH6ayn H. fl. fnr. h can., 1945, 1—2, 23—29.
PojeHUBUT T. 3. B kii.: PecJicpaTbt naytitux pa6oi /IciniiirpaacKaro
liHCTiiTyia nirHeiibi TpyAa 3a 1953 r, ,1., 1954, CTp. 148—152.
PB38KOB B. A. B kh.: npeae.nbno AonycTiiMwc KotmeitTpamm aT.«o-
ctjiepuux aarpasHciiHi'i. B. I. M, 1956. CTp. 26—39.
P » 3 a h o b B. A. P\'kcboactbo no xoMMyitajibiiofi rHriiene. T. I. M.,
1961.
Pusihob B. A. FHr. ii caH., 1961, 6, 3—8.
PasaiioB B. A. B kh.: Dpcae^iMio jionycTiiMbie Koitixeiitpamui aTMo-
c<$epHbix 3arpa3iiCHHH. B. 8. M. 1964, CTp. 5—21.
Pb3 a iiob B. A., ByuiTyesa K- A., Hobhkob JO. B. B kh.:
npeaeflbiio aonvcTsisiue KoiiueiiTpau»n atMoc<|>epnbix 3arpfi3HCHiii!.
, B. 3. M., 1957, CTp. 26-39.
C a a h a o b a M. C., C e ji * ii k h h a K. FI, UI t y p k ii h a O. K. r«r.
h can., 1965, 5, 11—15.
CmapcKH'H. B. B kh.: Matepiia.iy XIII naymioii ceccim CBepj-
AOBCKoro HHCTiuyTa nirHeitw Tpyaa h npo^namiornii. Caepiioacx,
1964.
CeflHHaH. A. Hir, it can,, 1962, 5, 41.
CeMeneHKoA. U,. Tnr. h can., 1963, 7, 49—55.
CeueneHKo A. iX . Ba/iauiesB. H., A p 3 a m a c u. e b E. B. B kh.-.
Bcec^K)3Haa Koii(J>cpeiiuitH no BonpocaM riinieHU Boaw n camiTap-
Hofi oxpaiiN BoaooMOB. Te3HCbi iioK-iaaciB. M., 1963, crp. 33—35.
C K m a k o b n B. Bonp niiTaniia, I960, 19, 6, 69—71.
C m it p ti o b lO. K. Hapyuienim noptfiipitiioBoro o6Mena npit 3adoJie-
BaHiiitx KepBHoft cncreMbi. ABTope4>. a>icc. M., 1953.
CMQJiHteB E. II. Apx. nat., 1961, 23, 5, 59—64.
C o n o m h it T. H. rnr. it can., 1961, 5. 3—8.
C n h p it h A. C. BitoxiiMitn, 1958, 23. 6, 656.
T h t a e b A. A , /I a p c k it ft 3. T., B o p it c o u a T. Fl, H a a e >k a «•
* a E. A. /laOop. Ae.io, 196-1, 4, 201—205.
T k a i H. 3 Titr. h can., 1965, 8, 5.
T»aieB FI. T. B kii.: npcACAbi'.o aonyctHMbie Koim.ci:Tpamui aTMo-
ctjepitbtx 3arpsi3ncmirt. B. 8. M , 1954, cip. 41—58.
y 6 a fi a. y n -i a c b P. Hir. it can., 1961, 7, 3—11.
y 6 s in y .1 Ji a e b P. JlaGop. Ae.to, 1934, 5, 300—302.
y .1 b it a 10. A\. Tcopan ii npaKTincn xpoiiaKCiiveTpti'ioBaiiiin. Jl., 1911.
y NitTe.ib H. JI., X a c m a h 3. ^1. Bcctii. AMH CCCP, 19Q4, 3. 23-36.
ii a a t o b a B. H Dir. ii can., 1DG2, 11, 4.
- 126 -
-------
Ouflimnoaa 3. X. P kh : MaTepiia^u iiavmioii Kon^icpcM'jim, hocbh-
uieiinori Donpocav: ninicnu Tpy,i3. npo4>;iaTo.iorini n nyovuu/icti-
HOli TOKCIlKO.IOnill B HClJlTRIIOl'l II HC1 C> IIM11'I OCKOl'l npOMUUMeilllU-
crn ycha, dp. 170—177.
H3iio;i. xcypii. CCCP, XXXIII, I, 1947,
17-27.
MhjkhkobB. A. B Kit.: npeae.ibno sonyciitMue KoimciiTpamui aTiio-
c4>cpnbix 3arpH3iienHit. B. 8. M., 1964. CTp. 21—40.
Ill h 6k as A. H. K yMemiio 06 oTpaB-ieiimi yKcyctioii kiic.iotoi'i ii.hi ee
scceHUiicfi. Kaaaiib. 1915.
"Ill n e ft (j> m a h O. A1. I"nr. it can., 1961, 5, 14—18.
lliTecce/ib T. A. apMaKo/i. h tokchkoh., 1941, 4. 3, SS—59.
Illy^bra T. M. B k»i.: FIpcae^ibHo aonycTHMue KoimeHTpamm aTMO-
c
-------
LITERATURE CITED IN 196 7 PAPERS
u) On-HCCittvuHon
Ar ec b a-M a i'i ko b a O. T. Bcctii. oTopsmaiip., 1951, 3, 3—8.
AfiseH6epr M. JX. PyccKan OTo.i.ipimro.ionifl., 1927, 5. 491—502 '
A kc »k A. 4>., By .1 u m e b f\ B. Frr. « c; h., J2, I9G2.
A^exceeBa M. B. Otii)sjc.U'»mc aTMOctpepiiux 3arpnanoiim". 1959.
A^iexceeea M. B OiipcAC-iemic aTMOC(j)fp!:iis sarpsuHcmift, 19oJ.
A/ieKce ea a M. B. ripeae-ibno JonyCTUMwe KOiiaeiiTpnu.r,: aivnz-
4>epHhix 3arpH3iieiUiH, 1964. s. 8, dp. 174—176.
A«inaTOB H. M. rwr. tpyja n npoc|>. aafSo-ieaaisna, 1964, 2, 14—IS.
A.ibaepH ZI. E. B kh.: Coape.Memiwe Bonpocu nepBiiava 3 ({unito-
jiotkh h naTo.ionni. M., .1955, cip. 41—46.
A^ibnepH JX¦ E. XojiHHepririesrKKe npoueccbi 2 naio-ioniii M.. I9C3.
AnaTODCKan B. C. Tc3i!cw .aohvtaaoB. MaTcpna.ibr irayiium u'toii
no TOKCHKofloriiii DbieoKOMO.ieKy.ijipiiyx ccc^itirciiMi':. iW-.;i.. il-Ct.
cip. 28—29
Ahuh JO. Jl. JIa6op. AC-10. 1964, I, 19—2!.
AhhikobC B. ["nr. h caH., 1952, 10, 7—12
Ah ox it ii n. K. npoS.ieMa ueiupa h nepit$ep;i;! a (t>H3;tanornii nepa-
HOH iaeHTe.1bHOCT!l ropbKHi't, 1935, CTp. 162.
Acc T. B., Bo.i B. B. h ap. Tpy.au n ¦MaTepaa.iu yKpatmcKoro ro-
cyaapcTaeHHoro HMCTHiyta pa6bneii vejutr.r.iN, 1926, ctp 11—55.
BafiKOB B. K. Tur. h can., 1953, CTp. 3—8.
DaHKOB B. K. B kh.: flpcjie.ibHo aonycniMbie xoimeiiTpaiiiiii amo-
ci}>epHUA 3arp»3iie!iiiH. 1964, ib. 8,.crp. 127—138.
Batca.ieHHiiK K. E. B kii-: Tes.icw 10-iV nayHHOii ceccm; Cucpi-
JiOBCKOro !ihctht>t3 riiriienu tpyaa ;i npoio)ia.ibiioi": naro.u-
rwi. Caepa-iOBCK, .1960, cip. 59—60.
E a km a it C. M. Jia6op. ae.io, 19^8, 5. 13—15.
BaKMaH C. M. Bpa>i. ae.io, 1950. 11. 110—105.
B a n a xo b c k h ft C. H. h B.i.ia xosckhh H. C. Mei«viu x-i-
• MHiecKoro -aHa-niaa KpoBii. M., 1953, ct?. 723.
BcJteHbKHH M. Jl. 3-1CMeHTbl KO.IimCCTOeilHOil C'UCHKII (f>ap::a!o.lM.
¦riwecKOro alexia. Haj., AM JlniBiiiioKOH CCP. Para, 195''
BecKOB C. /I. OciioBu .xiiMU'ieCKCfl Te.\!io.io?iii. Al, 1962.
CexTepeBa H. 11. h ycoa B. B. >i3i;c.i. >.>ypH. CCCP, I960,
46, I, 108—III.
BoHrpaA 3. M. n Ui.Tnn»u B. . Tur Tpy.ia h npoi>. sauo-
^toBaHHH, 1960, 2, 9.
B o h r p a a 3. M. h 1U .1 « n h ii B. . Te3HCbi .T;K.ia,ioB H.iyii'mi
KOHifepeimHii ropbKoscKoro fii!CTiiT>Ta r.ir,iu ipyaa « hporfi-
3d6o.icsan;ifi, tiocBnmeimue iiTorasi HaoiHO-.tcc.ieaoBaTc.ii.CKOii
pafioTbi HHCTHTyra 3a 1957 r, Fopi>K!3, 1958, CTp. 20.
BopuceHKOBa P. B. 3KcnepMMCHTa.ii>Hb:p ircc/KUOoamta no rt-
rHeiiKsecxoi'f xapnxTepiicTiiKc npo;i3BOACTwc.:j!Ofi :iu.ni (feppocina-
bob. /luce. xanA. M., 1952.
. Bop hcob a M. K- B kh.: npc.ic.iLiio jonycniMue KoirneHipamt.i ai
MOC(|)ep!ib]x 3arpH3iieiiitit,. 1960, d. 4, crp. 61—75.
ByuiTyeBii K. A. riirHCHH'iccK.m oac!i:
-------
Byiiitycoa K. A. B kh.; npi'jc.iMio aonycniMue Ktntui.iiTiia.uiii ;it-
>iio:(|>cpiii>i„ CeMeiieitxo A. H..
THr. H Cdii., I960, I, 57—61.
Bw.\obck3k C. At., T ii h j 6 v p r C. A., X a ,i« 3 o a a 0. H.
Mctojw onpe.vMemiH spejtmx EcmeCTB a iB03.nxe k jpyrux cpj-
aa\. H. 1. 1950, crp. 104-—107.
B a ii y .in k fl. Xiimiis MoitouepoB. T. I. Hep. c MciucKoro. rto.x
pea. anax H. Jl. KHyiiHxaa. Ii3j. HuotrrpaHHofi .iiucpaTypw. AV,
1960.
Be.unKO G>. K. B kh : KparKaa siiMiisecKaR sstmiK.'ione.aHa. M-,
J96I, T. I, CTp. 90—91. •
Bounap A. O. BttojioriHecKO * po.ib MiiKpo».ie.MeHT03 b o?ra!iii3Me
xiiBOTHux h «e-ioBeKa. At., 1953, CTp. 61—65.
Bo/iKOBa A. n. rnr. « caH , 1959, [, 80—82.
BcMKOsa H. H. 4>r.3j:o.i. xya>:. CCCP, 1951, 5, 691--693.
B o ji b $0 b c k a 8 P. H. ii .1 a b bi a o b a r. H. OSopimx Ha-.-'i-
•Hbix pa6oj 3a roan OTenecxacMiioii do'ihw. At, 1915, CTp. 155—15'J.
BpeaHbie iBemecTBa 3 npo\ibCKoro hhct;it\t:i i?m. Q. 0. Spucsima, 19-31.
reHAepceii, XarapA B kh : BpejHue ra3bi b npoMbiui.-iciiiio.-
cm M.—Jl., 19C0, cip. 1G9.
FiKec C. r. HepBHas oiicTexia 11 BHyrpeHima ceKpeiiim, 1955.
rei|>Tep Jl. H., liiyjib'Man E. H., fl o .i a x C A . riii.ib-
jw a n H. H., 3apeuKan A. II. 11 K.iefiH C. M. Tpv&u
BopoHe>kCKOro .vejiimiHCXoro iwcTJiTVTa, 1935. r. V.
P H^JieK A. 11 LUtepH E. S.ieKTpoitHbic cneKTpi>: norvromcHMii
oprammecKiix coeiiiiHeHHf!. Hep c aHr.i. not pea Jl. A. D.homci;-
' eJiwia. H3.fi. HiiocrpaHiiofi .itiTepaiypbi M., 1957.
rxpcKan E. fl. B kk.: Bonpocw mnieiiw Tpyaa, npo^naTtxnofiifii 11
npoMbiuj.ieHHofi HKTOKcifKauiiii. CBepa.iDBCK, 1959.
P Hte^uoK fl. H., TepuiKOB fl. A. 3p rrporpaMMu khk Memi
KfliiHHiecKoro Hcc.tMDBainiH Kpoaii. H3J CKdupcxoro OT/IC.ICIUIH
AAtH CCCP. KpacHoapcK. 195'J.
r ji a y 6e pat a 11 C. E. B Tpyjax Bopone>KCKoro Mej;m:nicKoro ii:i-
ctHTyTa, 1957, t. XXIX, crp 31—34.
I" 0 .1 m 11 H ck a a M. T. ;i3iio.r ;KvpK. CCCP, 1961, XVII, I, II.
roJty6eo'A. A. Tpyflbi h ayMKOu ceccini Jle.-tHiirpaacKoro ii!ictiitvt3
rwriieifbi ipy.ia ii-npcx+>3ai5o-ieR3nitfi, nocBHiaeuHKe uioraM pa6oTU
3a 1957 r. JI . J959, crp. 146—153.
roJibj6epr AI- C. CaniiTapiinn oxpaiia ar.MOc4>ep:ioro 303flv.xa.
At., 1951 .
Fo.ibACepr At. C. Caniirapiinn oxp^ia nr.Moc^epiroro B03.ty.\a iin-
ce-ieintbix mcct. At, I960,
l'opn Jl. 3. Fur tpyaa h npoipr.afio.iccannn, 1963. II. 44—51.
- 129 -
-------
r o p ui k c b C. H. O coapc.MeiiMux jictoj.tx ninieiiniccKiix hcc.ic.io-
BatiHH. M., 1964, cTp. 9—24.
PoijiMeK^ep B. A. Tur. m caH., 1900, 4 . 9—11.
f. o $ m ex.1 e p B. A. B Kit.: npc.ic.ibuo .ion>cTt!Mbic Ko;mciiT;>nun:i
3Txoc4>epiibix 3arp&3"e!Mfi. M., IC6I, d. VIM.
rotuea A. H. Bonp. Mex .mimiih, 1958, t. 4.
I'piiroposa O. n. Bonp. o.xp. suit. irjcr. 1953. 10, 50, 55
PpiiropoBa 0. in. Mexoc.iOBauKan iiMn.iTpiin, 1957, 2. 233.
TpiiropoBa 0. FI., Tiitob T. 11. Boiip. o.xp mst. i; acr, 1957
1. 31. 34.
T piiroposa O n. Banpocu -o.xp. mnr. ir ,ier., I9G2, 9, 13, 17.
rpHuinyH ^1.5. Bo-iuuiiie 303iuioepHhix 3arps3iteHHii. M., 1960, CTp.' 7—38.
Tycei M. H. B kh.: iripeae.iwio aonyrrimhie KOHueHTpamw awioc-
epHh»x 3arps3HeH;n1. Mearna. M., 1!>60, b. 4. ctp. 26.
ryces M. H. h Cmhphob 10. H. B kh.: .ripeae-ibHo AonycniMWC
KOHaeHTpaumi aTMOc<}>epHb« 3arp«neHnii. M.. 1960, b. 4, cto.
139-142.
Fvccb M. H., 4 e .1 n k a ii o b K. H. Tnr. >t can., 1963, 5, 3—8.
R >i n t p h e a B. R. Bkj.i.i. aKCnep. 5no.i. ii Mex, 1939, 8, 6.
H m ii t p.n e b a H. B. .TaSop. ae.io, 1^63, 6.
KrvHOB A. A. B c6.: K-niHnKO-ninieHntecKiie iicc.ieAOsanim no tok-
chhcokkm BcmecTaaM, npinieiiHcuyvj s iiobwx npoti3BoacTBax.
Jl., 1940, b. 2
CropoB A. n. Mop^o.ionmecKnn ana/ins Kpoan Mc.iriu. ,M., 1954.
EropoB K. B. TavAbt Acxpa.\nHCKoro .uea.mnnCKoro iiHcriiTyi;i,
1958, t. 14, CTp. 205
Ept'MHH B. n. Kaenapcxan 3. A. Bno.xit.Miin, 1950, t. in, u. 2
)K r y h n. B. JlaOop. Aeno, 1962, 4, 23.
yj ck a h P. M flpeae-ibiio aonycTiiMbie xoimeiiTpaunn haobh-
tmx napoB, raaoa it nu.iii « aoaayxe paoomtx ¦noMeweinifr. 1933,
b. 32, dp. 52. .
3 a k p jkc b ck i! ft E. 6., Bacit.inia Jl. V /IfOMiiuecuenTHaH
jiifKpoCKoniiH b K.i;tHitKO-reMaTo.iorit'iecKii\ itcc.iejoaastiiHX. MeA-
raa. M., 1963.
II m a uj e b a H, B. Hir. n can., 1963. 2, 3—8.
iluKosxi A. A. B iiHorpaaoB-1 B. A. Te3;icw aoiaaAoa no -
vhhoA ceccni! Hoboc;i6hpcko.-o iia\MHO-ncc.ieiOBaie-ibCKoro cainr-
iapiioro itiicniryTa 15—18 Maa 1966 r. HoaocnSnpcx, 1956.
CTp. 23—26.
Kaaac Jl. H. A?m:b o$Ta.ibMO.i»rnit, 1925. t. I. CTp 505—526.
Ka.iafiyiOB H. H. ycnexn coBp. 6»o.i.. 1940. 12, b 1
KatieHcxnii C. C. AVarepua.iw a.ih cjiapuaKO-iontu aueroijicsioiia.
/luce. aokt. ens. 1889.
KanxaeiB 3. A. fnr. m can., 1963, 12.
Kora h A. X. Bio.i.i. 3Kcncp. Gho.i. ii mcvu 1959, 4, 8, 10, 10D —
113.
Koran H. B. rio.iHp»rpn4>tiMsckiu'i aitn.tHS b npoMbiiu.icnnocaim-
tapnoii xnxtttn. Msanu. .M, 1961, CTp. 118—120.
- 130 -
-------
K 0 XC € B H li K O B B. A., Al C 1U C p C K i! fl P. M. CoflpCMClIH b!C MCTu-
flbi ann/noa s.ieKTpojime^a.iorpaMM. Mejriis'. M., 1963.
Kos.tob H. B, A3Tope4>epai aiiccepraumi Ha concKamie yxeHoii ere-
¦nen,i aoKtopa Mea;tai:iiCKiix Havx. Mhhck, 19G2.
KxiiioHOBa B. A., Akcchobs B. B. fur. n can., 1963, 7, 7—11.
KooCaKOBa A. H„ Kpe.\iOH Haynnofi cecrr.i
HHCTHiyia ninieHhi rpyaa h npo^aaSo.-icaaHsiH AMH CCCP, no-
CBBiueHHbie 40-.ieTHio Be.iiiKOi"i OKrR6pbCxoii couiia-nicTiPiecKoff rc-
BQjiioufitt, 16—21 neKaGpH. 1. HI, M., 1937.
KopoTKoea JO. K. Bccthiik OTopimo.iap., 1959, 3, 69—72.
KopoTKOBa 10. K. ripoMb:iii.T?:iHasi TOKCHKo.iorn». M, 1960, ct;>.
141—150. ,
KpaBKOB €. B. ii3;io.i. jkvp:i. CCCP, 1940, 23, 4, 313—322
KpacKHHa H. A., PyTpo-sa H. M. Tpyau Mocnoacxoro nay i-
•HO-HCc^eaoBaTe.ibCKoro micTiiTVTa anirjeMiio.ioniK h MHKpoCiio-
jiophm. HstMyno-iorim h npo({)ii.mKTiiKa K-.itue^uwx sui$eiC4Hii- 1962,
a. 9, cip. 180—186.
KpacoBMUKaa C. 3. ycnexii cospesieHHof: Ciio.ioniH, 1951, 32. 2,
166—192. 23 (H. H. Iilvpunm, C. O. Myp'inKoaa, H. A. Be.ioo,
1957):
K p a c o a h u k a n M. J1, 3 a n o p o ;k e u T. C. B kh.: MaTep.in.in •
Hay4.
JlaaapeB .H. B. Bpenibie BeiueCTBn a npoxbiiu.ieiiHoeTu, 1963.
/lapHOiiOB /]. O. runicua Tpyja w texHMKH OcaonacnocTii, 1954. V
/I e b h n C. H. B kh-.: Cscpj.ioscKan oG.mcrwafi xoM$epei!UHfi iicspo-
naTOJioroa, nciitnaTpoB h Hefipoxupypros, 1950.
/leBH«a 3. iH. CSopnux pa6or TOKCiiKg.iorn'iecKoft viaOop.iTop 11
iiHCTHTyra ninieKU Tpvja ii npac{>3a0o.ie3atu:i't, 1948, t. XII, .le-
HHHrpaji.
/lepiiep H. n., B p y c h .1 o b c k it fi E. C A.i.TepnmccKue 30111110-
({)H.ibiihie aaSo.icnaHHR. Kuea, 1961.
J1 k 111 3 h. B kh.: ripoje.ibno aonycriiMUe Ko;iiaeinpau'itn ar.Moci|»e?-
hux" 3arpH3iieii;ifl. B VII. M., 1963.
- 131 -
-------
J1 k> 6.1 ii a E. H. B kh.: apM.iKo.ioriis ;i TOKnisononm, 1950 13
3, crp. 33-37.
JlioQ.iJina E. H. B kii.: I1cc.ie.io3 .item b oS.iacTii npoMbfui.TMiiioii
TOKCUKo-iorim. 19-18, 12, 5, 74.
M a r h h u k ;i fi A. H. CyGopaiiHamia b Hepanoi'i circieMs it ee jiia-
leHHe b 4»!3;io.iorHH n naTo.ionm. M., 19-18.
MaKapos n. O. AieKsaT.-ina onrmecKas \po-iaxcHH a jjxuiio.ii-
thm k K.iKHHKe. /!., 1932. __
M a k a p m e r k o A. . H3MeHCinm Hep3iioi"i ci-rrcsibi npii i'i7ro>;c;t-
*aumi ¦MaprajiueM. Knea, 1956.
M a k c it x O'B iiM H- B. ^Kvpii. bucui. Hepa. aesT., 1954 4 G
909—SI 2.
M a p t bi-ii o b a A. H. Bonpocw r;irHciibr Tpysa b npOitjaoACTo; no-
bw.\ biuob cHHTenmeoKiix bo.ioko'.i {xanpoHoeoro h asiH.voro).
KaHi. jKCc., 1957. .
Ma pTU ho 8 a A. n. THr. xpy.ia » npoduaOo.ieaaHufi, 1957. 1
23—29.
Maraiiaesi E. H.. Tiir. ipyaa ir npo63£6o.ieBaH;iii, 1961, 7,
41—44.
MeficeflbM. H„ Cohosh k B. A. BuoifmiiiKa, 1956, r. 1.
B«n. 3, ctp. 262—274.
Me pko b JI. M. 06maa Teopiis :i Meroausa caHHTapHo-CTanicTii-
HeCKOro KCC-ieAooaHiiH. I-tax 2-e. M., 1963.
M h>1 /i e p C. B. B c6.: Ilpeje.ibiio jonyCTiriye xoiiueiiTpamui aT«o-
ctJwpHw.x aarpfUHe-niii. MeAri!3; 1955.
M k h a ei A. A. CSoptntK tpyaoa Kon4)epe:ijiiii mo.icuhx nirnein-
ctob 41 caHiiTapHbtx apaieii. M., 1965.
H. A. Tpyju naviHofi cecc;m .leHiiiirpa.icKoro nayiiiii-
MCMeflOBaTc.iLCKoro iiiicTinyTa ritr. Tp\_:;! n npoep3aGo.i..T.,-.n:iii.
nocBJimcimoii HTora>i paCoTw sa 1956 r. Jl , 1958, CTp, 237- 2-i3
' M h a u a k an mi A. B. B kii.: ripe.ic.ibMO aonycTiiMbic «omie:srpau:t'i
araoofiepiibix 3arpfl3ncsi:iri. M„ 1964, b. 8, CTp. 89—118.
M y p o a a h H a « C. H. C6op;-fi!K nayiHbix pafior Mockosckdto r,."5-
^acTHOro Hayimo-ncc^eiior.aTe.iboKoro caHinapHO-niriicriii'iccK r *
micTKTyTa. M., 1948, t. 1, cip. 82—124.
MyxHTOB B. M. Tiir. h can., 1962,-6, 16—24.
My.iHToa B. M. B kh,: flpejeijuo aonyCTiiMue Koiiue;trp.in.r:
aT«oci{)cpKbi.x 3arpH3iiciiin"i. B , VII, M„ 1903.
Muthiik II. OapMaKonoriiH n TcKCHKO.ioriiH, 1940, 3, 6, J9--,(i.
H a b p o u k ii i'i B. K. Bccth. AMI I CCCP, 1900. 3, 57—67.
Ha3apeHKO B. A. Tpyaw GnoreoMiMH'iecxoil Jiaflopa-ropitH All
OCCP. 1937, 4, 265 -271.
HexoTOpue npoS.ieMbi rurneiiw Tp)ua ;i npo$ecc;tona.ibiioA nn:r>-
jioniH. JIoa pea. 3. P. Miipys.uepa. Me.ini3. At., 1950.
HHKOJiiee H. M. Cod, mcj. 19>1. 18, 4, 38.
M B. B kii.: .Merojhi onpe.ic.icn-.i* Bpeaiibix aemccrn
B Bosjyxe h apyntx cpc.inx. 4. I. M.. 1969, crp. 238 —240.
Hobhkob 10. B. B kh.: ripcie.ibiio jonycT.tMbte xoMueiupaami ai-
iioc^epHhix 3arpn3;ieiiii'i, 1957, b. 3, -crp. 85—108.
O A o ui a ui b it .1 ir Jl. I'. Tur. n can.. 1962, 4, 3—7.
0.1« mi it e b a O. H , P e t ii e p B M., P y c mi o b a A. H B k;i :
Bonpocu rurueiiu Tpyja ii npo;|inaTO.ioriui np.i padoTax c 6^1130-
jiom h ero roMo.nornuii. HncniTyT ycoscpuie.'iCTaoaainm Dpaneii
«m. KwpoBa. Jl., 1958, b. 17, CTp. 8—22.
- 132 -
-------
0.1 b ui a ii c k h fi M. if. M n x a m e b a B. B. Tpyaw Pocra.TCKoro-
H3-floHy Me.iituiiiiCKoro imcnima. 1935, 2, crp. *120—127.
O p .1 o b a A. A., M a 3 y is n a a f. H. Cos. wca., I960, 9.
OCHOBll 3!!,l'JKpillic.lOIlll!. Iloj p€->, H. S\. /\pa3HIII!a, At. . M'.'-
pewiiHCKoro, 19S3.
II a B.i oe M. fl rio.inoe cofipamie Tpyjoi, 1949, t. 5, ctp. 112—137
h dp. 142—153.
ria.ibueB JO. II. Ti:r. n can., 1964. 2, 28—33.
fl a xo.\i bi h ee A 11. fur. h can.. I960, II, 77—84.
rieTpoa A. A. ycnexii xh«;ih. 1953, 22, b. 8.
rieTpos B. H. Dir. h ca*., 1950. 2, 60—62
n ct p y h b k ;i ii a A M. B kh.: ripaKTimecKan 6uoxiiMim. M.,'1951,
crp. 120.
II >i o t ii h k o b a M. M. B kh.: npcae.ibHo aonycTitMbie Konueiirpa-
uhh aTMOCcfiepHbix 3arpR3nemiS. Tloa pea. npo'ijj. B. A. Pn3ano3A,
1S60, .b. 4, CTp. 75—91.
SloKpoBCKJift A. A. iBoe>i. .siex "iKypii., 1953, 9, 61—65.
Fl o k p o e c k h fi B. A. ToKciiKo.tonin ii ruriiriia npoiiSBo.iCTBa cim-
TeTKiecKoro vavvywa. M., 1955.
rioKpoBCKKfi B. A. ; Bcecoioi:ioii ko :cpnoro eosjyxa, 1964.
PowKOBa B. M riaTO.iorii!!, k-ihhhkb h TepanKH OTpae-ieHiiii.
Hhcc. Jl., 4948.
Pokhubiit T. 3. Pe<£epaiw Kaynuux paOoi JlciimirpaACKoro mi-
CTHTyia nintenu Tpyja sa 1953 r. .H.. 1954, CTp. 148—162.
PoiiTOaK A. M„ Z1 c j a 6 p k ui o h jih U. M. AoK.ia.iw AH
CCCP, 1959. 124, .V? 4, CTp. 957.
Poiuhk H. B. Bpa-i ae.io, 1952, 9, 819—S24.
Pouik ii H. B. rwr. h can., 1952. 11, 4?--53.
P o in h m H. B Tiir. Tpy.ia h npo4>3a6o.ienaHiiii. 1962. 5. 17—22.
P o ui h m M B B kh.: ' ToKCMKO.tor,in pt'jKux xcta-l.iOB. M., 1963,
CTp. 83—95.
PycaKOBa A. B., F .1 a ] o b a O. H. K-hhi. >iex, 1943, XXf, 12.
P A 3 a h o b B A. Fur. ii can,, 1951, 6.3—8.
P r 3 a ii o b B. A. B kh.: npeie,u»no ..icmycriiMbii: KOir.ieiiTpniuni ar-
Hoc^epiibix 3arp»3iieiiiirt. Ms.t. «MeAmuinn», 196-1, b VII!
- 133 -
-------
Pa 3 hob B. A Pvko30Jctbo no KasrMyiin.ibifoft riiriiciie, 1961, t. I.
CTp. 158—159. *
P h 3 a ii o b B. A. ritr. ii can., 19-19, 5, 3.
P 8 3 3 ii o a B. A. Cainirapaasi ovpaiM anioctficpiioro uo3.iv\.i. 1951
P » 3 a hob B. A., B y in t >¦ e b a K. A., HobhkouIO. B B km.:
npejieuno jonycriiMbie KoiiiieiiTpauiiii arMocicpnux 3arpfi3iieit;iii
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CaaH/iofi3iio.Tortii! Tpyjn.
npofnaTO-iormi ii npoMuw.ieHHoi'i TOKC.iKo.iontit. CBepxioecK,
1962. t. VI.
CajiHJioBa AV C CfiopiiMK ypa.Tbcsoro $n.nia.tn AKaaeMiiit H:i-
yK CCCP, yp2.ibCKnff rocyjapcraeii'ibiH YimsepciiTeT. 1964.
Ce.mHK.Hiia K- II- Tur. n can., 1961, 10. 6—12.
Ce-Meitc iiko A. Zl. rur. it can., 1961, 5, 3.
CeneHtJiKO A. A- Fur. ii caH., 1963, 7, 49.
CesiCHeHKO A. A- B iMETepna.iax XIX Mockobckoh uaysiio-npaK-
tKnecKofi KOHiJjepeiimMi no npoS.ieuaM npOMbtime;inofi rnriieiiw
M„ 1-963.
C e m e h e H k o A. A IlepBaH no3o.iwcpcit-
UHH no HToraM iiaymiux HCMieaoBatmii 3a 1963 r. llH.-tMTsra o;V
inert ii KOMMyHa.nbHOM riirnetib< mm. A. If CwcHiia AMU CCCP. M .
1954, dp. 34-35
C e p r e a b O. C, K -i!' m e h k o A. A. BecTHHK penTrsHQ.iori:!i it pa-
aHOflorHM, 1957, 5, 76—81.
C e p e hc< h-P o h uj e h. BitoxitMii'ieoKite MeT0Jbi_ jstflriiOJa ii nc-
c-iejxoBaiiHB Mej. M3x Byxapecr. 1963, CTp. 259.
C e h e ii ob H. M.» Fl a b .1 o b H. B s e a c m c k it ft E. E. 3>ji-
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Cokojiod E. H. B c6.: OpiieimiposomifetH petf.ieKC it op;teimipa-
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C.i oh ii m A. R. B kh.: Ohut loyaium pery.umm'i i^ii;iio.iontiii'CKitx
. t^yHKUMH. Al.—J].. 1953, t. 2.
Co.io.vtHH T. H. Fitr. it can., 1961, 5. 3.
Co.iom hii T. M. B kh.; flpeic.tbito aonycTitMbie KoiiuciiTpaunit ariio-
c^epiiw^ 3arpst3i[emiii Al., 1962, b. 6.
Co.iomh ii T. H. rnr. it can., 1961, 3, 3.
C n m h y. E. H. B kh.: .Tnrireiia ii TOKC;tKO.ior:m hobuv nccTmtiMo::
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CrcK.iosa P. H. B c5.: OpiieiiTHpoBOntuft pcfi.icKC it opiieimtpo-
BO:iHO-Hcc.ieaoDnTC.ibCKaH jcsne.TbitocTb. Al, 1958, CTp. 183—191.
C t o b h o b c k it ft A. O. r»r. it can., 1961, 10, 70—72
CmatKHH B. M. B kh.: ITpcAe-ibtfo .tonycTUMux KoimetiTpamtu ht-
MOC^JcpHbi.x 3arpa3:t;iiiirt. Al., 1952, d. 6, crp. !)6—103.
- 134 -
-------
Ta.\Hpon sM. T. B x»i.: npeje.it.-io joaycTiiMMe KOHueJirpau«;t ar-
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T h y epoi:min no aonpocaM npouimvieHHoft tokciiko-
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Tkjud n. T. Fur. ii can, 1903, -1, 3.
ToAopoB H. K^iiHimecxne .laSoparopHue .:cc.iea,093H;if! a nei.i.ir-
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T p o n $. C. B kh,; Bonpocbi furkeHu Tpjua, npo;pecc-.ioHa.nt,-!:o,"i nn-
TO/ionm ii TOKCifKO-ionin b npOMWiu.ieiiiiccTjt CeepA-iozcKOii 06-
.laCTii, 1955.
y 6 a ft y .1.1 a e 3 P. B kh.: npeje.ibHO jonycnnrbie KOHi;eiirpau;i.;i
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.yfiaftay^^aeB P. Tiir. h can.. 1961, 7, 3.
y»apoBCKas O. At. flpofi.ieMbi 3HjOKp;i!io.ior;ui it ropMOitorepn-
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y ji » m -A. 10. M- Teopitn n npaxriiKa xpoiiaKCHMetpii;!. /I , 1941.
Tsiiioit h ¦:-ie«|>Tex:iMii,iecKOi> npoMuutoen-
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e .u> a m a. H 10. T. B jot.: npe.ie.ibiio aonycTUMhie Ko:ine>iTpau m
aTMOc^epHux aarpjuHeHisft. AV, 1962, e. 6, dp. 103—127.
4>o.mhh A. fl. C6opHiiK TpyaoB xou^epemum Mo.io.abix rnriiemi-
ctob h caM'itTapHbtx BpapeDA-iHH B. C. Tpyju BopowoKCKoro McawuHHCKoro Mitcri!T>T2
h oC^aciHofi caHiiTapHO-rtinieHimecKOf! .m6opaTopi;t3Ho.i. >xypn. CCCP, 1937, 22, I. 129—137.
3c xii it H. A, BuflaBCKan. P. Al. npoG.iv.MU 3iuoKpHho.io.~nii
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3ckhk H. A., M » i a 0 .1 o i H B. Ew.i.i. 3KCnep. Gho.1. h Mex,
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#poc:iaBCKafl P. H., P o 3 o b c k u fi H M. B kh.: Bto? -j: Bce-
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