PROPOSED
    WATER QUALITY
     INFORMATION
          Volume II
          OCTOBER 1973
U.S. ENVIRONMENTAL PROTECTION AGENCY
        Washington, D.C. 20460

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                          FOREWORD
     The Federal Water Pollution Control Act Amendments
require the Administrator of the U.S. Environmental
Protection Agency to publish both criteria for water
quality and information for the restoration and
maintenance of aquatic integrity, and the measurement
and classification of water [Section 3Ql*a(a)l and 2,
P.L. 92-500].

     Volume I of this two volume series contains the
criteria for water quality for the protection of
human health and for the protection and propagation
of desirable species of aquatic biota.  Volume II of
the series contains information on the maintenance
and restoration, measurement,  and the classification
of waters.  Also those pollutants suitable for maximum
daily load calculations are identified.

     Both Volumes I and II are published as proposed
documents with a 180 day period provided for public
comment*
                     I  Russell E. Train
                        Administrator

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                         Volume II
                          Proposed
               Information for Water Quality
              Environmental Protection Agency
                                                               Page


I.  Legislative Basis                                             1

II. Introduction                                                  2

III. Factors for Consideration for the Maintenance and
     Restoration of Integrity of Water and for the
     Protection and Propagation of Aquatic Life and
     Wildlife  304 (a)(2)(A)(P)                                    3

         Aluminum                                                 4
         Antimony                                                 4
         Arsenic                                                  5
         Barium                                                   5
         Beryllium                                                6
         Bismuth                                                  6
         Boron                                                    6
         Bromine                                                  7
         Cadmium                                                  7
         Chloride                                                 8
         Chromium                                                 9
         Copper                                                   9
         Cobalt                                                   9
         Fluoride                                                10
         Iron                                                    11
         Lead                                                    12
         Lithium                                                 12
         Nitrate                                                 13
         Nickel                                                  15
         Phosphorus                                              15
         Manganese                                               16
         Mercury                                                 16
         Molybdenum                                              16
         Selenium                                                17
         Silver                                                  17
         Uranium                                                 18
         Vanadium                                                19
         Zinc                                                    19


IV.  Measurement and Classification of Waters  304(a)(2)(C)      20

    A.  Measurement Techniques                                   21

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1.   Physical-Chemical Methods                           21

     Acidity                                            21
     Alkalinity
          Electro-metric Titration                       22
          Automated Methyl Orange                       24
     Arsenic                                            25
     Biochemical Oxygen Demand (BOD)
          Modified Winkler With Full Bottle             26
          DO Probe                                      27
     Organic Carbon  (Total and Dissolved)               28
     Chloride
          Titration                                     30.
          Automated Ferricyanide  Method                 31
     Colorinetry or Electrometric Determination         32
          Chlorine Requirement                          32
     Chemical Oxygen Demand  (COD)                       32
          Titration -  (High Concentration)              34
          Titration -  (Low Concentration)               35
          Titration -  (Saline Waters)                   36
     Color                                              37
     Cyanide
          Colorimetric                                  38
          Titration                                     40
     Dissolved Oxygon  (DO)
          Modified Winkler -  Full Bottle                41
          Probe Method                                  43
     Fluoride                                           45
     Hardness                                           46
     Total Hardness                                     47
     Metals  (Atomic Absorption)                         48
     Mercury
          Flameless Atomic Absorption                   51
          Cold Vapor Technique -  (Biological Materials) 52
          Gas Chromatography  - (In  Fish)                54
          Gas Chromatography  - (In  Sediment)            56
          Atomic Absorption  Spectroscopy
          for Surface  Waters                            58
     Methylene Blue Active Substances                   60
     iJitrogen                                           61
          Ammonia                                       61
               Colorimctry                              fil
               Titrinetric                              G3
               Automated  Colorimetric                   64
          Kjeldahl                                      65
               Titrimetric                              65
               Automated  Phenolate                      6G
               Colorimoteric                            68
          nitrite                      ,                 60
          Nitrate  and  Nitrite                          70

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               Automated Cadmium Reduction               70
               Automated Hydrazine                       72
          Organic plus Ammonia     .                      74
     Trisodium-tlitrilotriacetic Acid  (NTA)               75
               Zinc - Zincon                             75
               Automated Zinc - Zincon                   76
     Phosphorus                                          77
          Single Reagent .Method                          77
          Automated Single Reagent Method                79
          Automated Stannous Chloride Method             81
     Silica                               '               82
     Oil and Grease                                      83
     pi!                                                  84
     Phenolics                                           85
     Solids                                              87
          Dissolved                                      87
          Suspended                                      88
          Total                                          89
          Volatile                                       90
     Sulfate                                             91
          Colorimetry                                    91
          Automated Chloranilate                         92
     Sulfide                                             93
     Turbidity                                           94
     Temperature                                         96
     Threshold Odor                                      97
     Specific Conductance                                99
     Identification of Weathered or Unveathered          100
     Petroleum Oils                                      102
     Pesticides                                          103
          Electron Capture -  Has Liquid Chromatography  103
          Organochlorine Pesticides                      104

2.  Biological Methods                                   107

     Phytoplankton and Periphyton
     Cell Counts and Identification                      107
     Volume of Periphyton                                100
     Cell Counts and Identification of Phytoplankton     110
          Visual Observation                             110
          Visual Observation (Filter Method)             111
          Visual Observation (Counting Chamber Method)   112
     Cell Volume Estimates of Plankton and Periphyton    113
     Periphyton and Phvtoplankton (Species Connosition)  114
     Diatom Species Identification                       115
     Cell Counts and Identification of Phvtoplankton     116
     Chlorophvll a of Phytoplankton                      117
          In Vitro                                       117
          Spectronhometric                               118
     Zooplankton Volume and Species Identification       119

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              Iri situ Productivity of  Phytoplankton                   120
                   Radioactive Hethoc!                                 120
                   Oxycren Method                                      122
              Periphyton and Phytoplankton  Species Composition       124
              Chlorophyll a, h and c of  Phytoplankton and Periphyton 125
              Cell Surface Area of Phytoplankton                      126
              Diomass of Hacronhytes                                  127
              Algal r-rowth Potential                                  128
              Chlorophyll a of Phytoplankton  (Fluorescence)           130


         3.  nioassay Procedures                                      132

    B.  Classification of Uaters                                      162

V.  Constituents Suitable for Maximum  Daily Loading   304(a)(2)(D)     164

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               Information for Water Quality
I.  Legislative Basis

    flection 304 (a) (2) of. the "Federal Water Pollution
Control Act Amendments of 1972"  (86 Stat. 816; 33 U.S.C.
1314 1972), hereinafter referred to as the "Act", provides
that the Administrator  (EPA) shall, within one year of
enactment, (by Oct. 18, 1973), publish information:   (A) on
the factors necessary to restore and maintain the chemical,
physical and biological integrity of the aquatic
environment;  (B) on the factors necessary for the protection
and propagation of shellfish, fish and wildlife for all
classes and categories of receiving waters and to allow
recreational activities in and on the water;  (C) on the
measurement and classification of water quality; and  (D) on
the identification of pollutants suitable for maximum daily
load measurement correlated with the achievement of water
guality objectives.  The information shall reflect the
latest scientific knowledge.

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II. Introduction

    The national V7ater Quality Standards Program was
initiated with the passage of the Water Quality Act of  1965,
Sec 10 (c).  The Water Quality Standards are comprised of use
designations for eeich water body or portion thereof, water
quality criteria to support the use designations, and
implementation plans for scheduling the construction of the
necessary treatment facilities.  The designations by water
uses are protection and propagation of fish, aquatic life
and wildlife (fresh water and marine), water for public
water supplies, recreational, agricultural, and industrial
waters.  The Water Quality Standards prior to enactment of
FWPCA Amendments of 1972 were applicable to only interstate
waters and their tributaries.  The Act extends the coverage
to include all intrastate waters and the state standards
have since been, or are in the process of being revised
accordingly.

    The objective of the Act is to restore and maintain the
chemical, physical and biological integrity of the Nation's
waters.  The National goal, Sec 101(a)(l), is to eliminate
the discnarge of pollutants into navigable waters by 1985,
with an interim goal, Sec 101(a)(2), being to attain by July
1983, water quality which provides for the protection and
propagation of fish, shellfish and wildlife and for
recreation in and on the Nation's water.

    This Information for Water Quality is Volume II of a
two-volume publication.  It contains information on what is
found regionally in the Nation's water; on the measurement
and classification of water quality; and on the
identification of pollutants suitable for maximum daily load
measurement.  Volume I, under separate cover, is the
Criteria document.  The Criteria for Water Quality compiled
within Volume I consists of prescribed limits of
acceptability for pollutants, each followed immediately by
supporting scientific rationale.  The Information for Water
Quality is provided to assist the user of Volume I in his
application of the Water Quality Criteria.

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III.  Factors for Consideration  for  the  Haintenance  and
      Restoration of Integrity of V7ater  and  for  the  Protection
      and Propagation of Acru.atio Life  and  Uildlife

    Restoration and naintenance  of aauatic integrity
reauires the identification and  quantification of  the
elements or comnonents acting on the environment and an
understanding of the effect each has on  the  aauatic
environment.  Presented, in Table 1 are the ranges  and  mean
concentrations of inorganic pollutants found in  the  major
freshv;ater basins in the United  States.  Data are  arranged
by basins, and give the source of pollutants and dates data
vrere collected.

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Agent

Aluminum
             Sources

Natural

Weathering of rock
                                                                               Concent rat i on s
                                                                               in U.S. Waters
Man-Made

Industrial waste,
mine drainage, and
wash water from water
treatment plants
                                                                Basin Areas
                              Concentrations (tng/1)
New Jersey surface water
  1)  spring flow
  2)  summer flovi
Northeast Basin
North Atlantic Basin
Southeast Basin
Tennessee R. Basin
Ohio R. Basin
Lake Erie Basin
Upper Mississippi R. Basin
Western Great Lakes Basin
Missouri H. Basin
Lower Mississippi R. Basin
Colorado R. Basin
Western Gulf Basin
Pacific Northwest Basin
California Basin
Great Basin
Alaska Basin
                                                                                                   0.09-0.
                                                                                                   0.08-0.
                                                                                           Mean Positive:
                                                                                          33
                                                                                          31
                                                                                           0.028
                                                                                           0.022
                                                                                           0.117
                                                                                           0.030
                                                                                           0. 141
                                                                                           0.056
                                                                                           0.018
                                                                                           0.017
                                                                                           0.213
                                                                                           0.068
                                                                                           0.050
                                                                                           0.333
                                                                                           0.030
                                                                                           0.063
                                                                                           0.015
                                                                                           0.011
1968
1962-67
Antimony
                        Industrial effluents,
                        but is rapidly removed
                        by precipitation and
                        adsorption
                        Principal rivers of U.S.
                                                                                              None detected  1958-59

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                             Sources
Agent


Arsenic
                Natural
                Weathering of rocks
                                        Man-Made
                        Waste from industry
                        mining activity and
                        residues from
                        pesticides
                                                                                Concentrat ions
                                                                                in  U.S.  Waters
                                                                 Basin  Areas
                                                                               Concentrations  (mg/1)
Kansas R. at Lawrence
Kansas R. at Topeka

Kansas R. and Lawrence R.

Kansas R.

Northeast Easin
North Atlantic Basin
Southeast Basin
Tennessee R. Basin
Ohio R. Basin
Lake Erie Basin
Upper Mississippi R. Dasin
Western Great Lakes Basin
Missouri R. Basin
Lower Mississippi R. Basin
Colorado R. Basin
Western Gulf Basin
Pacific Northwest Basin
Great Basin
Alaska Basin
0.003
0.00&

0.002-0.010

0.002-0.008
                                                                                            Mean  Positive:
                                                                                                            0.031
                                                                                                            0. 017
                                                                                                            0.035
                                                                                                            0.050
                                                                                                            0.066
                                                                                                            0.308
                                                                                                            0.069
                                                                                                            0.037
                                                                                                            0. 123
                                                                                                            0.091
                                                                                                            0.053
                                                                                                            0.022
                                                                                                            0.068
                                                                                                            0.020
                                                                                                            0.034
1970
1970

1970
                                                                                                                   1962-67
Bariun
Weathering of rock
                                        Brines trom oil well
                                        waste and effluents
                                        from mining areas.
Northeast Basin
North Atlantic Basin
Southeast Basin
Tennessee R. Basin
Ohio R. Basin
Lake Erie Basin
Upper Mississippi R. Basin
Western Great Lakes Basin
Missouri R. Basin
Lower Mississippi R. Basin
Colorado R, Basin
Western Gulf Basin
Pacific Northwest Basin
California Basin
Great Basin
Alaska Basin

Coosa R. Below Rome, Ga.
                                                                                            Mean Positive:
        0.021
        0.025
        0.026
        0.025
        0.043
        0. 042
        0.039
        0.015
        0.063
        0.090
        0.060
        0.067
        0.027
        0.042
        0. OU1
        0.017
                                                                                                            0-340
                                                                                                         (max.  cone.)
                                                                                                                   1962-67

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                             Sources
Agent


Beryllium
                Natural
Weathering of
mineral beryl
                                        Man-Made
Effluent from atomic
reactors, metallurgy,
aircraft, rocket and
missile fuel indus-
tries
                                                                                Concentrations
                                                                                in  U.S.  Waters
                                                                Basin  Areas
                                                                               Concentrations  (mg/1)
Northeast Basin
North Atlantic Basin
Southeast Basin
Tennessee F. Basin
Ohio R. Basin
Lake Erie Basin
Western Great Lakes Basin
Missouri R. Basin
Pacific Northwest Basin

Monongahela R. at
  Pittsburgh, Pa.
                                                   Mean Positive:
   0.002   1962-67
   0.00012
   0.00005
   0.00016
   0.00028
   0.00017
   0.00005
   0.00023
   0.00002

   0.00122 1962-1967
(max.  cone.)
Bismuth
                Weathering of rock
                        Metallurgy; medicinal
                        Principal rivers of U.S.
                        streams in California
                            None detected
                                   0.0006-0.0008
          1958-59
Boron
                Weathering of boron-
                bearing rock
                        Industrial effluents
                        of weather prooting
                        wood, tire proofing
                        fabrics, manufacturing
                        glass and porcelain,
                        production of leather
                        and carpets, cosmetics,
                        photography, artifi-
                        cial gems, high
                        energy fuels, bacteri-
                        cides and fungicides
                        Northeast Basin
                        North Atlantic Basin
                        Southeast Basin
                        Tennessee R. Basin
                        Ohio R. Basin
                        Lake Erie Basin
                        Upper Mississippi R. Basin
                        Western Great Lakes Basin
                        Missouri R. Basin
                        Lower Mississippi R. Basin
                        Colorado R. Basin
                        Western Gulf Basin
                        Pacific Northwest Basin
                        California Basin
                        Great Basin
                        Alaska Basin

                        Colorado R. at Yuma, Ariz.
                                                                            Mean  Positive:
                                           0. 032
                                           0. 012
                                           0. 029
                                           0.02"4
                                           0.067
                                           0.210
                                           0. 105
                                           0.019
                                           0. 15"4
                                           0. 131
                                           0. 179
                                           0.289
                                           0.030
                                           0. 1<»3
                                           0. 08«
                                           0. 028
                                                                                                            1. 800
                                                                                                         (max.  cone.)
                                                                                                   1962-67

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                             Sources
Agent
Natural
Man-Made
                                                                                Concentrations
                                                                                in  U.S. Waters
Basin Areas
   Concentrations (rnq/1)
Bromine
                        RainfaLl, combustion
                        of leaded gasoline
                        containing ethylene
                        dibroroide, effluents
                        from industries pro-
                        ducing salt, chemical
                        medicinal compounds,
                        and disinfectants
                        Lake Superior, Mich.
                        Lake Superior tributaries
                        Lake Michiqan
                        Lake Huron
                        St. Clair 5.
                        Lake St. Clair
                        Detroit R.
                        Lake Erie
                        Lake Ontario
                                   0.007-
                                   0.005-
                                   0. 01 1-
                                   0.013-
                                   0.019-
                                   O.OU5-
                                   0.020-
                                   0.020-
                                   0.038-
              0.033
              0.260
              0.021
              0.029
              0.039
              0.055
              0.028
              0.05U
              0.077
      1969
Cadmium
Weatherinq of rock
Effluents from indus-
tries using cadmium
such as metallurqy,
electroplating,
Northeast Dasin
North Atlantic Basin
Southeast Basin
Ohio R. Basin
Mean Positive:
                                        ceramics, piqmentation. Lake Erie Easin
                                        photography, nuclear
                                        reactors,  insecticides
                                        and antihelminthics
                                                Upper Mississippi R. Basin
                                                Western Great Lakes Basin
                                                Colorado R. Basin
                                                Western Gulf Basin
                                                Pacific Northwest Basin
                                                Great Basin

                                                Cuyahoqa R. at Cleveland,
                                                Ohio
0.005
0.003
0.005
0.007
C.050
0.006
0.005
0,002
0.010
0.005
0.001
                                                                                                            0. 120
                                                                                                         (max.  cone.)
1962-67

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                             Sources
Agent
                Natural
                                                                               Concentrations
                                                                               in  U.S.  Waters
                                        Man-Made
                                                                Basin  Areas
                                                       Concentrations  (nxj/1)
Chloride        weathering of rock,
                aniir.
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                             Sources
Agent


Chromium
                Natural
                                        Man-Made
                                                                                Concentrat ions
                                                                                in  U.S. Maters
                                                                Basin  Areas
                                                                              Concentrations  (mq/1)
                        Industrial effluents
                        using chromium such
                        as metal pickling anJ
                        plating operations,
                        anodizing aluminum,
                        leather processing,
                        manufacture of paints,
                        dyes, explosives,
                        ceramics, paper, glass,
                        corrosion inhibitm-ii
                        and many others
                        Tennessee R. Basin
                        Ohio R. Basin
                        Lake Erie Basin
                        Upper Mississippi R. Basin
                        Western Great Lakes Basin
                        Missouri R. Basin
                        Lower Mississippi R. Basin
                        Colorado R. Basin
                        Western Gulf Basin
                        Pacific Northwest Basin
                        California Basin
                        Great Fasin
                        Alaska Basin
                                                   Mean Positive:
0.006
0.007
0.012
0.007
0.006
0.017
0.016
0.016
0.025
0.006
0.015
0.0014
0.009
                                                                                                   1962-67
                                                                St.  Lawrence  R. at Massena,
                                                                  N.Y.
                                                                                                            0.112   1962-67
                                                                                                         (max.  cone.)
Copper
Weathering of rock
Corrosive action ot
water in copper pipes,
industrial effluents
such as discharged
from textile mills,
pigmentation, tanning,
photography, engraving,
electroplating, in-
secticides, fungicides
and in many other in-
dustrial processes
                        Northeast Basin
                        North Atlantic Basin
                        southeast Basin
                        Tennessee R- Basin
                        Ohio R. Basin
                        Lake Erie Basin
                        Upper Mississippi R, Basin
                        western Great Lakes Basin
                        Missouri R. Basin
                        Lower Mississippi R. Basin
                        Colorado K. Basin
                        Western Gulf Basin
                        Pacific Northwest Basin
                        California Basin
                        Great Basin
                        Alaska Basin
                                                                                            Mean Positive:
0.015
0.017
0.01U
0.011
0.023
0.011
0.01U
0.007
0.017
0.019
0.010
0.011
0.009
0.012
0.012
0.009
                                                                                                                   1962-67
Cobalt
Weathering of rock
Industrial effluents
from plants usinq
cobalt such as metal-
lurgical firms, cer-
amics and glass manu-
                        Northeast Basin
                        North Atlantic Basin
                        southeast Basin
                        Ohio R. Basin
                        Lake Erie Basin
facturintj, the tungsten Upper Mississippi R. Basin
carbide tool industry   Western Great Lakes Basin
and others              Missouri R. Basin
                        Lower Mississippi R. Basin
                        Colorado R. Basin
                        Pacific Northwest Basin
                                                                Allegheny  R.  at  Pittsburqh,  Pa.
                                                                                            Mean Positive:
                                                                                                            0.01t
                                                                                                            0.009
                                                                                                            0.001
                                                                                                            0.019
                                                                                                            0.033
                                                                                                            0.018
                                                                                                            0.011
                                                                                                            0.008
                                                                                                            C.036
                                                                                                            0.011
                                                                                                            0.008
                                                                                            O.OU8
                                                                                         (max.  cone.)
                                                                                                                   1962-67

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                                                             10
Agent           Natural
                                         Man-Made
                                                                                 in U.:;.  .-.a'-ers
                                                                  hasin Areas
                                                                                                 Concent r if i or.:-
Fluoride        Weathering  of rock
                containing  fluoride
Industrial effluents
from plants  usinq
fluorides  in the
production ot  steel,
disinfectants,  pro-
served wood  arid
muci]a<.jes, cjldSE and
enamels, chemicals
It is aiiued  i-o many
public water supplies
to reduce  incidoi.ce
of cavitios  in t.et-th.
                                                                                                 0. 1
                                                                                                 o. u
                                                                  surface waters of
                                                                  Drinkinq water of  1t>3
                                                                                 atfas in
                                                                  HocKy Creek an^i Ejst
                                                                    Gallatin R.  (various
                                                                    locations alnnq rivers
                                                                    f rciii sources ot pol I'.jric
                                                                  Colorado R. Basin  (;nax  at
                                                                    K. iielow Littlef ield , Ariz.)
                                                                  Pacific Slope Basins  in Calif.
                                                                    (max. at colusa Trough  riear
                                                                    Colusu, cilit.)

                                                                  Great Basin excluding c;redt
                                                                    SalV Lake (max. at  !!umbolii-
                                                                    V. near Kye patch,  "Jev. )
                                                                  Paciric Slope Basiro  in
                                                                    w.isnincjton and Upne.r
                                                                  Coluirhia R. Basin  (max.
                                                                    at OruL Creek near Smyrna,
                                                                    kashi nqton)
                                                                  Pacific Slope Basins  ir. Ore.
                                                                    anri Lower Columbia 'A. Basin
                                                                    (max at V>dllii Walla K.
                                                                    near Touchet, Wash.)
                                                                  Snake R. Basin  (max. at J5nake
                                                                    P. at Kinq Hill, Idaho)
                                                                  Alaska  (max. at Tonsina K.  at
                                                                    Tonsina, Alaska)

                                                                  North Atlantic Slope  Oasins
                                                                    (max. at Wissachickon cre°k
                                                                    at Fort Washington, Pa.)
                                                                  South Atlantic Slope*  Basins
                                                                    (max. at Alafia R. at Lithia,
                                                                    Kla.)

                                                                  Ohio R. Basin  (max. at Tus-
                                                                    carawas R. at Newcorrerstcwn,
                                                                    Ohio)
                                                                  -St. Lawrence K. Bacin  (max
                                                                    at Black R. =it Elyria,
                                                                    Onio)
                                                                                                                Cct.  TJf. 1
                                                             U.a-U.9

                                                             J.1-0.1
                                                             ). 1-0.7
                                                             o.o-i. a
                                                             0. 0-17-0
                                                             C.O-U. 2    1964-t>S
                                                             0.0-1.7

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                                                             11
                             Sources
Agent
                Natural
                                                                                Concentrat ions
                                                                                in  U.S.  Waters
                                                                BdSin  Areas
                                                                                               Concentrations  (mq/1)
                                                                Missouri  R.  Basin  (nax  at
                                                                   Cedar Creek  near Columbia,
                                                                   Mo.
                                                                Hudson P. and  Upper  Missis-
                                                                   sippi R.  Basins  (max  at
                                                                   Souris  H.  near Verendrye,
                                                                   N. Dak.)

                                                                Lower Mississipfi R.  Basin
                                                                   (max at Center Creek  at
                                                                   Oronooo,  Mo.)
                                                                Western Gulf of Mexico  Basins
                                                                   (max at Double Mountain Fork
                                                                   Brazos  R.  at Justiceburq,
                                                                   Tex.)
                                                                Illinois  surtace waters
                                                                  at  Illinois R. at
                                                                  Peoria-1961)
                                                 (max
                                                            0.0-3.U    1964-65


                                                            0.0-1.2




                                                            0.0-15.0


                                                            0.0-2.1




                                                            0.0-2.U    1956-66
Iron
                         ot iron
                salts from soil and
                rock into both
                qround and surface
                waters
Aciu mine dr.iinaqe,
the effluents from
the many industries
usinq iron in their
processing,  and the
oxidation ot iron used
in such items as
ships, automobiles,
refriqerators and
many others
Northeast Easin
North Atlantic Basin
Southeast Basin
Tennessee R. Basin
Ohio R. Basin
Lake Erie Basin
Upper Mississippi R. Basin
Western Great Lakes Basin
Missouri R. Basin
Lower Mississippi R. Basin
Colorado R. Basin
Western Gulf Basin
Pacific Northwest Basin
California Basin
Great Basin
Alaska Basin

Sabine R. near Ruliff, Tex.
                           Mean Positive:
0.051
0. 019
0. 120
  .037
  .028
  .035
  .035
  .022
  ,037
  .069
0.010
0. 173
0.032
O.OU6
0.070
0.025
                                                                                                            0.952
                                                                                                         (max.  cone.)
                                                   1962-67

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                             sources
                                                                               Concentrat ions
                                                                               in U.S. Waters
Agent


Lead
                Natural
Weathering ot lime-
stone and galena
                        Man-Made
Corrosion ot leau
in pipes and near
industrial outf
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                                                            13
                             Sources
                                                                                Concentrat ions
                                                                                in U.S.  Waters
Agent


Nitrate
                Natural
                       tecal nidtoridl
                                        Effluents  trcm  plants
                                        producing  cl;emical
                                        fertilizer.  Applica-
                                        tion or chemical
                                        fertilizers  on  agri-
                                        cultural l.inds  that
                                        are then loached  into
                                        the surface  ami perco-
                                        latinq ground waters.
                                        Application  of  ferti-
                                        lizers on  residential
                                        lawns and  gardens.
                                        Municipal  treatment
                                        nlant effluent
Easin Areas
                               Concentrations  (roq/1)
nurtace waters of 98 U.S. rivers

Surface waters of U.S.
Range: 0.1-10.1
                                            0.5
                                            3.2
                                            1.1
Missouri R. water
  in Omaha, Neb.
  in Kansas City, Mo.
  in St. touis County, Mo.
  in St. Louis, Ho.

Hudson R.

Iowa

Ohio, 10» of municipal water  supply

Mahoninq R.

Ohio rural water supplies

Drinkinq water in 163 metropolitan
  areas in U.S.

Missouri wells

Lake Erie

Colorado R. basin (max. at Gila F.
  below Gillespie Dam, Ariz.)
Pacific Slope Basins in Calif,  (max.
  at Salinas R. near Sprechles, Cal.)
Great Basin excluding Great Salt  take
  (max. at Jordan R. at Salt  Lake
  City, Utah)

Pacific Slope Basins in Washington
  and Upper Columbia R. Basin (max.
  at Flett Creek at Tacoma, Hash.)
Pacific Slope Basins in Oregon and
  Lower Columbia K. Basin  (max. at
  V»alla walla R. near Touchet, Vlash.)
  1960-

Aug. 19
Sept 19
Oct  19
     19
                      1951-
        0.2-8,6 monthly
        1.8
        1.C-9.0    "
        2.1-11.3   "
                                                                                                            0.2-1.0

                                                                                                            10.0-100.0

                                                                                                            1.0

                                                                                                            10

                                                                                                            10.0-100.0

                                                                                                         Average:  2.3


                                                                                                            •5-300

                                                                                                            1.0

                                                                                                            0.00-60.0

                                                                                                            0.0-38.0

                                                                                                            0.0-17.0



                                                                                                            0.0-12.0


                                                                                                            0.0-4.2
                         19

                         19

                    July 19

                   March 19

                         19

                      1950-

-------
                                                           14
                             Sources
Agent
                Natural
                                        Man-Made
                                                                                Concentrations
                                                                                in U.S. Waters
                                                                 Basin Areas
Concentrations  (mq/1)
                                                                 Snake R.  Basin (max.  at Palouse R.      0.0-11*.0
                                                                   at  Hooper,  Wash.)
                                                                 Alaska (max.  at Trail R. near Lowing,   0.0-2.8
                                                                   Alaska
                                                                 North Atlantic Slope Basins (max. at    0-0-62.0
                                                                   Rockaway R.  at Pine Brook, N.J.)
                                                                 South Atlantic Slope and Eastern Gulf   0.0-620.0
                                                                   of Mexico Basins (max. at Planta-
                                                                   tion Rd. Canal near Fort
                                                                   Lauderdale,  Fla.)

                                                                 Ohio R.  Basin  (max.  at Great Miami      0.0-23.0
                                                                   R. at Elizabetbtown, Ohio)
                                                                 St..  Lawrence R.  Basin (max. at          0.2-113.0
                                                                   Black R. at  Elyria, Ohio)

                                                                 Missouri R. Basin (max.  at Horse        0.0-77.0
                                                                   Creek near Vale, S- Dak.)
                                                                 Hudson R.  and  Upper Mississippi R.       0.0-302.0
                                                                   Basins (max. at Blue Earth R. near
                                                                   Rapidan, Minn.)

                                                                 Lower Mississippi R.  Basin  (max. at     0.0-302.0
                                                                   Fountain Creek at Pueblc, Colo.)
                                                                 Western Gulf of  Mexico Basins  (max.     0-0-5U.O
                                                                   at Yegua Creek near Somerville,
                                                                   Tex.)

                                                                 Illinois surface waters  (max.  at        0.0-«8.i»
                                                                   Illinois R.  Meredosia-1962)

                                                                 Lehiqh R.  Easin  (max. at Black Creek    0.0-23.0
                                                                   near Weatherly-1963)
                           196U-
                           1964-
                          1956-6


                          1945-6

-------
                                                               15
                             Sources
Aqent


Nickel
Natural
Man-Made
                                                               Concentrat ions
                                                               in U.S. Waters
Basin Areas
                                                New Jersey surface waters
                                                   1) spring flow
                                                   2) summer flow

                                                New Jersey surface waters
                                                   Big Flat Brook
                                                   Calcareous - site  1
                                                   Acidic - Site 2

                                                Lower Mississippi R. Basin:
                                                   Californiaa Gulch  and
                                                   Arkansas H. at Malta, Colo.
                                                   (max. at California Gulch)
   Concentrations  (mq/1)
                                                                                                  0.002-0.009
                                                                                                  0.001-0.017
                                                                                                  0.006
                                                                                                  0.001

                                                                                                  0.0-0.069
                                                                                                                        1968
                                                                             196«-65
Phosphorus
The decomposition of
orqanic matter
Siltation.  Applied
fertilizer.  Effluents
of plants makinq
fertilizers, rr.atciies
armaments, orqanic
chemicals, ortho-
phosphoric acid, foo.i
supplement in animal
foods, water soften-
ers, anu in metalurqy
Northeast Basin
North Atlantic Basin
Southeast Basin
Tennessee R. Basin
Ohio R, Basin
Lake Erie Basin
Upper Mississippi R. Basin
Western Great Lakes Basin
Missour R. Basin
Lower Mississippi R. Basin
Colorado R. Basin
Western Gulf Basin
Pacific Northwest Basin
California
Great Basin
Alaska Basin
Mean Positive:
O.OU4
O.OU8
O.OU3
0.042
0. 130
0. 153
0.2U3
0.031
0. 353
0.081
0. 121
0, 173
O.OU7
0.083
0.037
O.OUO
1962-67

-------
                                                                16
                             Sources
Agent
                Natural
                                        Man-Made
                                                                               Concentrations
                                                                               in U.S. Waters
                                                                Basin Areas
                                                                                              Concentration:.
Manganese
Leached from rock
into groundwaters
Mining effluents from
from industries using
manganese in the
production of steel
alloys, dry cell bat-
teries, qlass and
ceramics, paints
and varnish, inks,
matches, fireworks
and in agriculture
to nourish manganese
deficient soils
Northeast Basin
North Atlantic Basin
Southeast Basin
Tennessee R. Basin
Ohio R. Basin
Lake Erie Basin
Upper Mississippi R. Basin
Western Great Lakes Basin
Missouri R. Basin
Lower Mississippi R. Basin
Colorado R. Basin
Western Gulf Basin
Pacific Northwest Basin
California Basin
Great Basin
Alaska
                                                                                           Mean Positive:
0.0035 1962-67
0.0027
0.0028
0.0037
0. 232
0. 138
0.0098
0.0023
0. 0138
0. 009
0.012
0.010
0.0028
0.0028
0.078
0.0018
Mercury
Leaching of rock
and volatilization
Effluents to air
emissions from chlor-
alkali plants, ap-
plications of pesti-
cides, fuel burning,
catalytic processes,
ore refining, sewage
treatment, waste
incineration, phosphate
rock, processing, paint
manufacture, and use and
breakage of mercury-
containing devices, and
many others
Ranqe of Hg found in all of
  the States in Oct 6 Nov. 1970
                                                                                                   0.5 ug/1-6.9 ug/1
Molybdenum
Mineral weathering
Mining and processing
molybdenum ore.
Effluents ot indus-
tries using molybdenum
in their operation
sucli as metallurgical,
glass, ceramics,
pigment producr.ion,
fertilizers and
other
Northeast Basin
North Atlantic Basin
Southeast Basin
Tennessee R. Basin
Ohio R. Basin
Lake Erie Basin
Upper Mississippi R. Basin
Western Great Lakes Basin
Missouri R. Pasin
Lower Mississippi R. Basin
Colorado R, Basin
Western Gulf Basin
Pacific Northwest Basin
California Basin
Great Basin
Alaska Basin
                                                                                           Mean Positive:
0.025
0.033
0.015
0.025
0.070
0. 068
0. 088
0. 028
0. 083
0. 095
0. 130
0. 02«
0. 030
0. PU5
0. 1U5
0. 017
                                                                                                                  1962-67

-------
                                                              17
                             Sources
Agent
Natural
Man-Made
                                                               Concentrat ions
                                                               in U.S. Haters
                                                basin Areas
                                                      Concentrations (mrj/l)
Selenium
                        Traces in ettltients
                        from plants usinq
                        silver to produce
                        sucn items as pig-
                        mentation tor paints,
                        dyes, glass, com-
                        ponents in rectifiers,
                        semi-conductors
                                                                Selected drinking water supplies in:
                                                                Connecticut                                0.010
                                                                Maine                                      0.010
                                                                New Hampshire                              0.010
                                                                Vermont                                    0.010
                                                                Delaware                                   0.010
                                                                New York                                   0.010
                                                                Pennsylvania                               0.010
                                                                Kentucky                                   0.010
                                                                Maryland                                   0.010
                                                                North Carolina                             0.010
                                                                Virginia                                   0.010
                                                                W. Virginia                                0.010
                                                                Alabama                                    0.010
                                                                Florida                                    0.010
                                                                Georgia                                    0.010
Silver     weathering of rocks
                                        Trace amounts in
                                        effluents trom in-
                                        industrial plants
                                        using silver such as
                                        electroplating plants,
                                        food and beverage
                                        plants, photographic
                                        chemicals producers,
                                        ink manutacturing,
                                        antiseptics for
                                        medicinal purposes
                                                Northeast Easin
                                                North Atlantic Pasin
                                                Southeast Basin
                                                Ohio R.  Basin
                                                Lake Erie Basin
                                                Upper Mississippi R. Basin
                                                Western Great Lakes Basin
                                                Missouri R. Basin
                                                Lower Mississippi R. Basin
                                                Colorado R, Basin
                                                Pacific Northwest Basin
                                                Great Basin
                                                Alaska Basin
                                                   Mean Positive:
0.0019 1962-67
0.0009
O.OOOU
0.0021
0.0053
0.003U
0.001U
0.0012
O.OOU3
0.0058
0.0009
0.0003
0.0011

-------
                                                             18
                             Sources
Agent:
Natural
Man-Made
                                                                                Concent.r jt ions
                                                                                in U.S.  Waters
                                                E'asin  Areas
                                                       Concentrations
Uranluai    Weathering of rack
                        Miniricj and proces-
                        sing of uranium in the
                        manufacture of atomic
                        weapons, the produc-
                        tion of radiojsotopes
                        in piles and reactors,
                        use of radioisotopes
                        in medical therapy,
                        scientific research
                        photography, and in
                        many chemical proces-
                        ses
                        Walkt-r K. Cilit.
                        Weber S., Utan'
                               Cottonwood Creek.,  Utah
                                 Creek,  Idaho
                        lllyt- Mountain, Calif.
                        Hell-Arches Naticnai Monunent,  Utah
                        Slot Sprinq-Pyramid  Lake,  Nevada
                          Truchee R. , Nevada
                        Walker H., Nevada
                        Humi;olt  R,, Nevada
                        Carson H., Nevada
                        Sevier R., Utah
                        Lake Tahoe, Calif.
                        Pyramid  lake, Nevada  (South  end)
                        Pyramid  Lake, Nevada  (North  end)
                        Walker Lake
                        Mono Lake

                        Principal rivers of U.S.

                        North Atlantic slope Basins  (3dx  at
                        Susquehanna R. at Harrisburq,  Pa.
                        and Potomac R. at Hancock, Md.)

                        St. Lawrence R. Basin
                          Genessee R. at Rochester,  N. Y.

                        South Atlantic Slope Basins  (max  at
                          Yadkin K. at Yadkin college,  N.C.
                          and St. Johns R.  at Christmas,  Fla)

                        Gulf of Mexico Basin  (max at
                          Missouri R. at Nebraska City, Neh.)

                        Colcrado R. at Yuma, Ariz.

                        Pacific Coast Basins  (max at
                          San Joaquin H. at Vernalis,  Calif.)

                        Yukon R. at Rampart, Alaska

                        Red R.  (N) at Grand Forks, N.D.
   n.0009
   0.0013
   0. 00?
   0.0003
   0. Ci, OOU
   0.OC^C
   o.oooos
   0.0046
   0.016
   O.G072
   O.GOU7
   0.0081
   0.0008
   0 . 0 U.
   O.C25
   0.077
   0. 139

none detected  1V5H-59

0.0004-0.0006  1960-61
                                                                                                            G.OOOtt
                                                                                                        19*0-61
                                                                                                         0.000<»-0.0007  1960-61



                                                                                                         0.0001-0.0006  1960-61


                                                                                                            0.007';,

                                                                                                         0.0004-0.008


                                                                                                            o.oom

                                                                                                            O.C01U

-------
                                                           19
                             Sources
Agent
     Natural
Man-Made
                                                                    Concentrat ions
                                                                    in U.S. Waters
Basin Areas
   Concentrations (mg/1)
Vanadium   weathering of rock
                             Leachinq of soils.
                             Degradation of plants
                             and animals. Effluents
                             from plants making
                             steel, glass, mordant
                             used in dyeing and
                             printing fabrics,
                             and other products
                        Northeast Basin'
                        North Atlantic Basin
                        Southeast Basin
                        Tennessee R. Basin
                        Lake Erie Basin
                        Upper Mississippi R. Easin
                        Missouri R. Basin
                        Lower Mississippi R. Basin
                        Colorado R. Basin
                        Western Gulf Basin
                        Pacific Northwest Basin
                        California Basin
                        Alaska Basin
                           Mean Positive:
                0.009
                O.U12
                0.010
                0.022
                0.05'i
                0.020
                0. 171
                0.025
                0. 10?
                0.009
                0.013
                0.030
                0.012
            1962-67
Zinc
Weathering of rock
Mining and processing
of ore.  Effluents
from plants making
Pharmaceuticals, dyes,
insecticides, and many
other products
Northeast Basin
North Atlantic Dasin
Southeast Basin
Tennessee R. Basin
Ohio R. fiasin
La k e Er i e
Upper Mississippi R. Basin
Western Great Lakes Basin
Missouri P. Basin
Lower Mississippi R. Basin
Colorado R. Basin
Western Gulf Basin
Pacitic Northwest Easin
California Basin
Great Basin
Alaska Basin
Mean Positive:
0.09f>
0. OU9
0.052
0.028
0. 081
C.205
0.045
O.C2"
0.039
0.085
O.OM
0.092
C.CUO
0.016
19b2-67
                                                                                                           C.02fi

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                                 20
IV.  Measurement and Classification of Waters   304 (a)(2)(C)

     A.  Measurement Techniques

     Methods included within the following section cover a
variety of menstirenents and techniques.  For some
pollutants, several methods are cited for measurement or
detection.  Each method is summarized and presents a
synopsis and is not intended to give a detailed
presentation.  Rather an outline of the technique is
followed immediately by the pertinent references for an
extensive review of the procedure.

         The techniques are divided into three  areas;
physical-chemical, biological, and bioassays.   It is not
within the scope of this document to detail precisely or to
recommend a specific method.  Often circumstances such as
sources and mixtures of pollutants, cost of test, and skill
of the personnel performing the test will dictate the
preferred procedure.

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                               21
                          ACIDITY
Principal Detection Techniquet  Titration

Summary of Method;  Sample is titrated to a final pH of 8.3.
Results reported as mg of CaC03 per liter.

Limitations;

    Interferences;  Hot applicable to acid samples from mine
drainage

Statistical Characteristics;           Over Observed Range of;
(By 40 analysts in 17
laboratories)

    Accuracy;           Bias              Acidity, as mg/1 C

                       +2.77%                      20
                       +0.52%                      21

    Precision:          Standard Dev.
                        1.79 mg/1 CaCOy            20
                        1.73                       21

    Time of Measurement (Maximum frequency, recovery period,
    etc.); Not stated, but fast.

Data Outputs;  Analog signal displayed on meter.

Special Sampling Requirements  (Collection, Storage,
Handling);  Refrigerate sample at 4*C.
Maximum holding time is 24 hours
References;

1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Annual Book of ASTM Standards, part  23  (Method
    designation D1067).  1970.  Society  for Testing and
    Materials.  Philadelphia, Pennsylvania.

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                                 22


                         ALiCALIMIYY


Name; of 'loasuronent Method;   Eloctronctric Titratiou
Principal Detection Technique;  Eloctrometric

Purpose of Measurement  (Important Applications);  Uriakiny
waters,ambient surface waters,' domestic and industrial
wastes, and saline waters.      - '

Summary of Method;  Unaltered sample is titrated to an
electromotrically determined end point of pil 4.5.  Sample
must not be filtered, diluted, concentrated, or otherwise
altered.

Limitations;

    Range of Applicability;  All concentrations

    Interferences;  Salts of weak acids, oils and greases

    Pitfalls; Special Precautions;   Analyze sanple as soon
    as possible after collection, preferably within a few
    hours.


Statistical Characteristics;           Over Observed Range of;

    Accuracy;           Bias              Alkalinity in rag/1 CaCU,

                       +22.29%                     9
                       - 3.19%                   113

    Precision;          Std. Dev. ,  nig/1 CaCQ^

                         1.4                       9
                         5.28                    113

    Time of Measurement  (-Maximum frequency, recovery period,
etc.);:Hot stated, but rapid.

Data Outputs;  Meter  (analog voltage)

Special Sampling Requirements  (Collection, Storagef Handling^;
Refrigerate" at 4°C.  Maximum holding time 24 hours.

-------
                                 23
References;

1.  Methods  for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center ,
    Cincinnati, Ohio.

2.  Annual Book of ASTM Standards, part 23 (Method
    designation D1067).  1970.  Society for Testing and
    Materials.  Philadelphia, Pennsylvania.

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                                     24
                      Total Alkalinity


Name of Measurement Method;  Methyl Orange

Principal Detection Technique;  Titration with colorimetric
detection of end point.

Purpose of Measurement  (Important Applications);  Surface .
waters,saline waters.

Summary of Method;  A Technicon Autoanalyzer is used,
employing methyl orange as indicator.  This is dissolved in
a weak buffer at pH of 3.1, just below the equivalence
point.  Addition of alkalinity causes loss of color
proportional.  Color measurement made by colorimeter at
approximately 550 nm.

Limitations;

    Range of Applicability;  10 to 200 mg/1 expressed as


Statistical Characteristics;

    Precision;  Std. dev. was ±0.5 mg/1 CaCOj  (in one lab)
using concentrations of 15, 57, 154, and 193 mg/1
    Time of Measurement  (Maximum frequency, recovery; period,
etc/H  30 min. warm-up, about 1 min. per deterrnintataon

Calibration Requirements;  Requires preparation of standard
curve of peak heights vs. concentration

Data Outputs;  Strip recorder  (analog voltage)
Special Sampling Requirements  (Collection, Storage, Handling)
Refrigerated at 4*C; sample analyzed as soon as possible.
References;

1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Technicon Autoanalyzer Methodology.  1961.
    Bulletin 1261.

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                                  25
                          ARSENIC

Name of Measurement Method^  Diethyldithiocarbamate method

Principal Detection Technique;  Colorinetry

Purpose of Measurement  (Important Applications);  Most fresh
water

Summary of Method;  Arsenic in sample is reduced to arsine,
AsH5 , in acid solution.  The arsine is scrubbed to remove
sulfide, and is absorbed in a solution of diethyldithio-
carbamate dissolved in pyridine.  The red complex thus
formed is measured in a spectrophotometer at 535 nm.

Limitations:
    Range of Applicability;  At or above 10 ug/1 As.   (If
    arsenic is organically bound, consult Standard Methods,
    13th Edition).

    Interferences;  High concentrations of chromium, cobalt,
    copper, mercury, molybdenum, nickel, or silver.

    Pitfalls; Special Precautions;  Difficulties may be
    encountered with certain industrial v/astes containing
    volatile substances.  High sulfur content of wastes may
    exceed removal capacity of lead acetate scrubber.

Statistical Characteristics;  In 46 laboratories:

                                       Over Observed Range Of;

    Accuracy;  Relative error = 0%          40 ug/1

    Precision;  Relative Std. Dev. = 13.8   40 ug/1 as As

Data Outputs;  Chart or meter (analog voltage)

References;

1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  UFA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Wastewater.  1971.  13th Edition.  American Public
    Health Association, Washington, D. C.

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                                   26


              BIOCHEMICAL OXYGEN DEMAND  (BOD)


Name of Measurement Method;  Modified Winkler with Full
Bottle Technique.

Principal Detection Technique;  Titration

Purpose of Measurement (Important'Applications);  Ambient
surface water, domestic and industrial waste waters
(especially sewage treatment plant effluent).

Summary of Method;  Sample of waste, diluted as appropriate,
is incubated for 5 days in darkness at 20eC.  The reduction
in dissolved oxygen concentration during this period yields
a measure of the biochemical oxygen demand.

Limitations;  Sec summary for Dissolved Oxygen  (Modified
Winkler Method).

Statistical Characteristics:
    Precision;                         Over Observed Range Of;

    Seventy-seven analysts in fifty-   An unspecified range with
    three labs analyzed samples of     mean value of 194 mg/1 BOD
    natural water plus an exact
    increment of biodegradable com-
    pounds.  At a mean of 194 mg/1
    BOD, the standard deviation was
    + 40 mg/1

Time of Measurement  (Maximum Frequency, Recovery Period, etc.);

Five to six days per determination.

Comments by Users;  Because of local conditions, types of
samples to be tested, and variabilities of the bioassay
procedures, no specific standard test for BOD has been
selected by EPA.

References:
1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Wastewater.  1971.  13th Edition.  American
    Public Health Association, Washington, D. C.

-------
                                    27
              BIOCHEMICAL OXYGEN DEMA1JD  (BOD)

Hame of Measurement Method; DO Probe

Principal Detection Teclinigue;  Electronetric method.

Purpose of Measurement  (Important Applications):  See
summary of BOD determination using Modified Winkler Method.

Summary of Method:  See BOD determination using Modified
Uinkler Method.

Limitations;   (See Summary for Dissolved Oxygen  (Probe
Method)).

Statistical Characteristics;  See DO determination using
Modified Winkler Method.

Comments by Usors^  Because of local conditions, typos of
samples to be tested, and variabilities of the bioassay
procedures, no specific standard test for BOD has been
selected by EPA.

Data Outputs^  Electrical signal displayed on meter.

Special Sampling Requirements (Collection, Storage, Handling)
Refrigerate at 4W C.  Maximum storage tine - G hourj.

References;

1.  Methods for Chemical Analysis of !7atcr and Wastes.
    1971.   EPA National Lnvironmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Uater and
    Uastcwater.  1971.  13th Edition.  7unerican
    Public Health Association, Washington, D. C.

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                                     28
            ORGANIC CARBON  (TOTAL A1JD DISSOLVED)

Principal Detection Technique;  Infrared spectres copy

Purpose of Measurement  (Important Applications);  Ambient
surface waters,domestic and industrial wastes,saline
waters; used for assessing potential oxygen-demanding load
of organic matter.

Suramary j3 f Method;  A micro sample of the wastewater to be
analyzed is injected into a catalytic combustion tube wnich
is enclosed by an electric furnace thermostated at  950 C.
The water is vaporised and the carbonaceous material is
oxidized to Carbon dioxide  (CO^.) and steam in a carrier
stream of pure oxygen or air.  The oxygen flow carries the
steam and COjr. out of the furnace where the steam is
condensed and the condensate removed.  The CO-j., oxygen, and
remaining water vapor enter an infrared analyzer sensitized
to provide a measure of CO2.  The amount of COX present is
directly proportional to the concentration of carbonaceous
material in the injected sample.

Limitations:
    Range of Applicability;  1 to 140 mg/1 total carbon.

    Lnterforonces;  Carbonates, bicarbonates

    Pitfalls; Special Precautions;  Since sample is injected
    into apparatus by syringe, the needle opening liuits the
    size of particles reaching combustor/detector.

Statistical Characteristics:
Twenty-eight analysts in twenty-one laboratories analyzed
distilled water solutions containing exact concentrations of
oxidizable organic compounds, with the following results:

Known Carbon      Precision as                     Accuracy as
Cone, as TOC,     Standard Deviation            ijias,       Bias,
mg/liter          TOC, mg/liter                  %        mg/liter

    4.9                 3.93                  +15.27      + .75
  107                   8.32                  + 1.01      +1.0J


Time of "easureraent;  Hot stated; assumed to be rapiu  (several
samples per hour).

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                                     29
Special Sampling Requirements  (Collection, Storage, Handling);
None indicated.  Maximum holding time - 7 days.

References:;

1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Wastewater.  1971.  13th Edition.  American
    Public Health Association, Washington, D. C.

3.  ASTfl Standards.  1970.  Part 23, Water,  Atmospheric
    Analysis.

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                                   30
                          CHLORIDE

Principal Detection Tochnic^ue;  Titration

Purpose of "loasurcnent  (Important Applications) ;   Drinking
v;aters, ambient surface v/aters.  Domestic and industrial
wastes, saline waters.

Summary of Method;  Dilute mercuric nitrate solution is
added to an acidified sample in the presence of mixed
diphenylcarbazone - bromophenol blue indicator.  The end
point of the titration is the formation of the blue-violet
diphenylcarbazone complex.

Limitations:
    Ran geof Applieability:  All concentration ranges of
    chloride

    Interferences;   Sulfites.  If presence is suspected,
    oxidize with hydrogen peroxide.

Statistical Characteristics;  By 42 analysts in 13 laboratories

    Accuracy;  Percent bias ranged     Over Observed Range Of:
    from 13.50 (at 10 rag/1) to         17, 18, 91, 97, 392,  398
    1.10 (at 398 mg/1).                mg/1

    Precision;  Standard Deviations    Same
    ranged from 1.32 mg/1  (at 13 mg/1
    C1-) to 11.3 mg/1  (at 393 mg/1 Cl~)

Data Outputs;  Visual observation and manual recording (or
may use automated titration, yielding an analog voltage
recorded on chart).

Special Sampling Requirements (Collection, Storage, Handling);
None indicated.  riaximum holding time - 7 days.

References:
1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and Waste-
    v;ater.  1971.  13th Edition, American Public
    Health Association, Washington, D. C.

3.  ASTM Standards.  1970.  Part 23, Water, Atmospheric
    Analysis.

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                                    31
                                CIILORID^

Mane of Measurement Mothodj  7uitorated rcrrioynnic'e Method

Principal Detection Techniqiie:  Titratior.,  usincr colorireter
Purpose of  Measurement (Inportant Applications) :
wa t drs"~ dones't'i c~ aridl"Tndus'trla!" wa"ste"&r~~sallne" w
   Ambient  surface
waters.
S urinary of Method:   Thiocyanate ion  (f?C?T)  is liberated fron nprcuric
tHi^>cyana'te~ th'ro'ucfh sequestration of rorcury by chloritV ion  to  forn
un- ionized mercuric chloride ion.   In  the  presence of. ferric  ion the
thiocyanate  SC11 forris highly colored ferric thiocyanate in
concentration  proportional to the original chloride concentration.

The deternination nay he perfomer1  autonaticallv usina a ^echnicon
AutoAnnlyzer cnployinq a colorineter with  380 nn filter for enclpoint
detection.

Limitations :

    Ranqe_of_ ApplJ.cahiJ.^i^tyj  1 to 250  rg/1 chloride

^»f-3-f*:if:fJ_Chara5^cri£>J:icR:            Over Observed Ranee Ofj

In a single  laboratory:

    PrecisJ-onj   Standard deviation      1;  100; and 250 pa/1
    +T)~'3~ncf71~ chloride                  chloride
        -j  Hen_svirenent:  About 2
    ninutesrpe'r sar,pl'e~'"v7ith 30 nin.
    warn-up.
             .   AnalofT voltaqe, recorded  on strip chart

§PP52-f J- ^J'l.'PlJ-I1^. Re3i! j-jr1*1^!1^5  (Collection, Storaae, Hand lino) :
Hone .  Tlax"inum~Tic)17l'ln~r time" -"V clays"

References :

1.  Methods  for Chemical Analysis of Water and Wastes.
    1971.  EPA  National Environmental  Research Center,
    Cincinnati,  Ohio.

2.  O'Brien, J.E.   1962.  Automatic Analysis  of
    Chlorides in Sewaqe.  Waste Engineer ina,  33:
    670-672.

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                                     32
                         CULORITir

Principa_l_ Detection Techniaue:  Colorimetry  or Electronetrio
DeTiernination

Prrnose of_ Measurement  (Inportanjt  Applications) :   To determine the
anount of chlorine to T>e add'ed" to  sample  to  acTiieve certain desired
results, such as control of colifom  densitites,  destruction of
certain chcnical compounds, or  establishment of specified chlorine
residuals .

The nethod given here is applicable to  drinkina water, surface waters,
donestic and industrial wastes, and saline water.

Sunmary of Method:  A solution  of  known chlorine content is added
incrementally ^;o a series of  sanple aliauots.  At the end of the
stipulated contact tine or when the desired  result has been achieved
the residual chlorine is measured  by  the  appropriate nethod.

Limitations :

    Range of Applicability:   Not single,  specific procedure, but
    varies wTEn purpose" or" result  to  be achieved.

    Pj.t_fa_lls_;_Rpecia2_ Precautions:  T7hen  the purpose is to obtain a
    speci"fTe7]'"c'nl.orine~'resl7lual~~'the  sane nethod of chlorine
    neasurencnt should be used  for operational control and laboratory
    testing.

    All pertinent infornation should  lie included in report of results,
    such as:  conditions of chlorination  (p!I, temperature, contact
    tine) ; nethod used for detemining  the desired result; and the
    chlorine renuired to produce result.

Statistical Characteristics:

    Accuracy_and_Precisipnj   Mature of  this  test precludes the use of
    accuracy an7i~precTslon~ statenents .
               Hot given, huh assumed  to  be r.nalocr voltage displayed
o"n ne"ter~o""chart .

References :

1.  Methods for Cher.ical Analysis  of Hater and Wastes.
    1971.  EPA national Environmental  Research Center,
    Cincinnati, Ohio.

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                                     33
2.  Standard Methods for Exanination of  Water and
    Wastewater.   1971.   13th Edition. American  Public
    Health Association,  Washington,  D. C..

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                                      34


                     CHEMICAL ox-rnr::i  DEMAND (COD)


Principal Pet ect ion^ Techn igue ;   Titratior - (Iliqh  Concentration)

Purpose of Measurement  (^Important  Applications) :   To determine
quantity o"f~oxygen required" to" oxT7!ize~ aTI~~org an ic natter ir a
wastewater sample under  specified  conditions.

Sunnary of Method:  Organic  substances  ir the  sample are oxidized by
po"tass'iui\r "eTchromate in  50%  sulfuric  acid solution at reflux
temperature.  Silver sulfate is  used  as  a catalyst and mercuric
sulfate is added to remove chloride interference.   The excess
dichromate is titrated v/ith  standard  ferrous annoniun sulfate, using
orthophenanthroline ferrous  corplex as  an indicator.

Limitations :

    Ranqe of Applicability:  Organic  carbon concentration of 15 rng/1
    "•-"••• ^" *f •" T •• m •• •• ^ ^^ •• ^•^•^^ » ™ * — •
    or higher.

    Interferences;  Chloride concentration over 2000 ng/1

Statistical Characteristics :

    Preci^io^i:  Eighty-nine  analysts:  Standard deviation of +27.5
    ng71~COD at knov/n value  270  ng/1  COD.
Special Sar.pl ing Rcguirenents^ ^Collection ,  Storaae , n.
Adcl "5" nT~ Kj SO^ per liter  oT^sanple"   MaxlnTirrT s'to'rage~lIne~-"
7 days.
References :
    Methods for Chenical Analysis  of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

    Standard Methods  for Examination of Water and
    Wastewater.   1971.  13th  Edition.  American Public
    Health Association, Washington,  D.  C.

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                                 35
                CHEMICAL OXYGEN DEMAND  (COD)

Principal Detection Technique^  Titration -  (Low
Concentration)

Purpose of Measurement  (Important Applications); Ambient
surface water,domestic and industrial wastes with low
oxygen demand characteristics.

Summary of Method;  Similar to that for COD-High Level,
except (1)that extreme care is exercised during sample
acquisition and handling and during analysis to insure that
no organic contaminants are introduced from glassware,
atmosphere, etc.,  (2) that highly pure reagents are used,
and (3) that chlorides are removed by complexing with
mercuric sulfato.

Limitations:
    Range of Applicability;  5 to 30 mg/1 COD

    Interferences;  Organic contaminants; chlorides

    Pitfalls; Special Precautions;  Volatile materials may
    be lost during sulfuric acid addition step.

Calibration Requirements;  Standardize reagents daily.

Special Sampling Requirements  (Collection, Storage, Handling)
Use glass sample bottles.  Preserve with IUSO«j.  Test
biologically active samples soon after acquisition.

References:
1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Wastewater.  1971.  13th Edition.  American Public
    Health Association, Washington, D« C.

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                                 36
                CHEMICAL OXYGEN DEMAND  (COD)

Principal Detection Technique;  Titration -  (Saline Waters)

Purpose of Measurement  (Important Applications):  COD
Determination in saline waters.

Summary of Method;  Organic and oxidizable inorganic
substances in an aqueous sample are oxidized by potassium
dichromate solution in 50 percent (by volume) sulfuric acid
solution.  The excess dichromate is titrated with standard*
ferrous ammonium sulfate using orthophenanthroline ferrous
complex  (ferroin) as an indicator.  Mercuric sulfate is
added to complex the chlorides during digestion.

Limitations:
    Range of Applicability:  Minimum of 250 mg/1 COD when
    chloride concentration exceeds 1000 mg/1.   (The removal
    of chlorides by MgSq, may not be complete in the case of
    strong brines.)

    Interferences;  Extraneous organic matter

Special Sampling Requirements (Collection, Storage, Handling);
Tf
Use glass bottles if possible; preserve with

Reference:
1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

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                             37


                                COLOR

Principal Detection^ Technigue_t  Visual  sensing

Purpose of Measur ement  ( Import an t_ Appl icat ions ) : Measures  color of
water derlveef f rom naturally occurring^ihaterials , e.g.  vegetable
residues such as leaves, bark, humus, etc.

Summary of Method;  Color is measured by visual  comparison of  the
sample with" platinum-cobalt standards.  One unit of color  is that
produced by 1 mg/1 platinum in the  form of the  chloroplatinate ion.

The Spectrophotometric  and Tristimulus methods  are useful  for
detecting specific color problems.  The usr of  these methods,  however,
is laborious and unless determination of the hue, purity,  and
luminance is desired, they are of limited value.

Limi t.at_ions_:

    Interferences!  Turbidity; highly colored industrial wastes.
    ?itlfiisi. Special S-?~-u--°"-s  Biological activity may change
    color" characteristics a? ter" sample is acquired.

C a 1 Ibr a ti pn_ Regu j.r erne n ts j  See Reference

Data Outputs^  Usually a visual observation, manually recorded.

Special Sampling Reguirements  (Collection. Storage, Handling) :
Store sample atT*C.  Maxlmum"Tiold"ing time -~^3" hours"!     """"
1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Wastewater.  1971.  13th Edition. American Public
    Health Association, Washington, D. C.

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                               38
                          CYANIDE
Principal Detection Technique;  Colorinetric

Purpose of Measurement  (Important Applications); Ambient
surface water,domestic and industrial wastes, saline
waters.

Summary of Method;  The cyanide as hydrocyanic acid  (HCIJ) is
released from metallic cyanide complex ions by means of a
reflux-distillation operation and absorbed in a scrubber
containing sodium hydroxide solution.  The cyanide ion in
the absorbing solution is then determined by volumetric
titration or coloriraetrically.  The colorirnetric measurement
employs the pyridine-oyraxolone reaction in which the
cyanide is coupled with free chlorine to form cyanogen
chloride and then with the pyridine to a glutaconic
aldehyde.  The aldehyde then reacts with l-paenyl-3-iaethyl-
5-pyraxolone to form a highly colored blue dye.

Limitations:
    Range of Applicability;  Cyanide concentrations below 1
    mg/1(sensitiveto about 0.5 ug/1).

    Interferences;  Sulfides, oxidizing substances.

    Pitfalls; Special Precautions;  Material tested may be
    highly toxic and should be treated accordingly.  Sample
    should be maintained at a highly basic pH.

Statistical Characteristics:
    Determination by 47 analysts yielded:

    Precision:     Standard Deviation    Known CM Concentration of;
                     0.020 mg/1                  0.02 rag/1
                     0.306 mg/1                  1.10 mg/1

Data Outputs;  Analog signal  (meter)

Special Sampling Requirements  (Collection, Storage, Handling);

       minimum sample size -  1 liter
       adjust pH to 11 at time of sample collection
       (using sodium hydroxide)
       analyze as soon as possible after collection
       (maximum holding time  - 24 hours)
       store at 4°c.

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                            39
References;

1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

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                              40


                           CYAi:ii)E

Principal Detection Technique:  Titration

Purpose of ileasuror.ient  (Important  Applications);  Ambient
surface water,domestic and industrial  wastes,Saline
waters.

Summary of Ilethod;  The cyanide as  hydrocyanic  aciu (IICII)  is
relt.asod from metallic cyanide complex  ions  by  means  of a
reflux-distillation operation and  absorbed  in a scrubber
containing sodium hydroxide solution.   Tne  cyanide ion in
tho absorbing solution is  then determined by volumetric
titration or colorirnetrically.  Tho  titrinetric ineasurenent
uses a standard solution of silver  nitrato  to titrate
cyanide in the presence of a silver  sensitive indicator.

Limitations:
    Range of Anplicability;  Concentrations  of  cyanide
    exceeding 1 mg/1

    Interferences;  Sulfidas, oxidizing  substances

    Pitfalls; Special Precautions;  See  summary for  Cyaniuo
    Colorimetric

Statistical Characteristics:
    Results by 47 analysts:

    Precision:     Standard Deviation    Know;:  CIJ  Concentration of
                       0.035 mg/1                 0.02  mg/1
                       0.333 mg/1                 1.10  nig/1

Data Outputs;  "lot stated.   (Probably visual  reading,
recorded manually).

Special Sampling Requirements  (Collection  Storage,  Handling);
See summary for Cyanide, Colorimetric.

Reference:
1.  Methods for Chemical /Analysis of Water  and Wastes.
    1971.  EPA national Environmental Research Center,
    Cincinnati/ Ohio.

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                           41
                   DISSOLVED OXYGKM  (1)0)

Uarte of Measurement Method!  Modified Winkler - Full Bottle
Technique

Principal Detection Technique;  Titration

Purpose of Measurement  (Important Applications); Most
ambient surface waters,most waste waters(but not donestic
sewage).

Summary of Method;  The sample is treated with manqanous
sulfate, potassiun hydroxide and potassium iodide and
finally sulfuric acid.  The initial  precipitate of manganous
hydroxide combines with the dissolved oxygon in the sample
to form a brown precipitate, manganic hydroxide.  Upon
acidification, the manganic hydroxide forms manganic sulfate
whicn acts as an oxidising agent to  release free iodine from
the potassium iodide.  The iodine, which is
stoiciiiometrically equivalent to tiie dissolved oxygen in the
sample is then titrated.

Limitations:
    Interferences;  Oxidizing or reducing materials
    (especially sulfites, thiosulfates, polythionates,
    chlorine, hypochlorite) nitrate ions, ferrous ions,
    organic matter, high concentrations of suspended  solids.

    Pitfalls; Special Precautions;  Sample may contain  low
    concentrations of ferrous ions  (less than 1 mg/1) and
    nitrates.  High concentration of either interferes.

Statistical Characteristics; Exact data not available,  the
following are approximate.

    Precision;  Reproducibility of 0.2 rag/1 of DO when  known
    concentration is 7.5 rag/1 DO.

    Time of Measurement;  Not stated, but a complex
    procedure.

Commonts by Users; Most common interferences overcome by use
of DO probe.

Data Outputs;  Visual observation or analog signal.

Special Sampling Requirements (Collection, Storage, Handling);
Grab sampling acquisition procedures are specified;also
preservative reagent to be added.  Analysis should be completed

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                               42
within 4-8 hours after acquisition of sample.

Reference^

1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

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                                 43


                   DISSOLVED OXYGEN  (DO)

Name of Measurement Method;  Probe Method

Principal Detection Technique;  Electrometric

Purpose of .Measurement  (Important Applications); Ambient
surface waters, domestic and industrial wastes.

Summary of Method;  The most common instrumental probes for
determination of dissolved oxygen in water arc dependent
upon electrochemical reactions.  Under steady-state
conditions, the current or potential can be correlated with
DO concentrations.  Interfacial dynamics at the probe-sample
interface are a factor in probe response and a significant
degree of interfacial turbulence is necessary.  For
precision performance, turbulence should be constant.  The
probe method may be used under any circumstances as a
substitute for the modified Winkler procedure provided that
the probe itself is standardized against the Winkler method
on samples free of interfacing materials.  The electronic
readout meter for the output from dissolved oxygen probes is
normally calibrated in convenient scale  (0 to 10, 0 to 15, 0
to 20 rag/1, for example) with a sensitivity of approximately
0.05 mg/1.

Limitations:
    Interferences;  Sulfur compounds and certain reactive
    gases(e.g., chlorine) may interfere.  Dissolved
    inorganic salts affect performance, but usually can be
    compensated for.  pH variation interferences with some
    probes.

    Pitfalls; Special Precautions;  Probes may be sensitive
    to temperature and may require temperature compensation.

Statistaical Characteristics;  Manufacturers'  Claims:

    Accuracy;  j^ 1% of true DO value

    Precision;  0.1 mg/1 repeatability

Time of Measurement;  Mot stated, but rapid.

Calibration Requirements;  Not stated.   (Should be
calibrated against standards with approximately the same
concentration of suspended solids as the sample).

Data Outputs;  Electrical signal displayed on moter

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                                44


Special Sampling Requirements  (Collection, Storage, Handling)
Hot stated.  Can provide continuous,  in  situ measurements.

Reference:
1.  Ilethods for Chemical Analysis of Water and Wastes.
    1971.  EPA national Environmental Research Center,
    Cincinnati, Ohio.

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                                 45


                          FLUORIDE

Maine of Measurement Method;  Automated Complexone Method

Principal Detection Technique;  Photometric, automated
(Teclmicon AutoAnalyzer)

Purpose of Measurement  (Important Applications);  Ambient
surface water,domestic ana industrial wastes, saline waters.

Sunraary of Ilethod;  Fluoride reacts with red cerous chelate
of alizarin complexone.  A positive color is developed and
intensity is measured by a colorineter at about 650 nn.
Teclmicon AutoAnalyzer is used.

Limitations:
    Range of Applicability;  0.05 to 1.5 rog/1 fluoride

    Interferences;  Aluminum

Statistical Characteristics;
    Precision;  At one laboratory, standard deviation of
    measurement was 0.018 mg/1 F when actual concentrations
    were 0.06, 0.15, 0.55, and 1.03 mg/1.

    Tine of Measurement;  12 samples per hour after 30
    minutes wanning.

Data Outputs;  Analog electrical signal recorded on
stripchart.

Special Sampling Requirements (Collection, Storage, Handling)
Not stated.  Can provide continuous, in situ measurements.

Reference:
1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

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                                46

                          HARDNESS


Principal Detection Technique;  Titrntion

Purpose of Measurement  (Important Applications);   Drinking
waters, ambient surface waters, domestic and  industrial
wastes.

Summary of Method;  Calcium and magnesium  ions  in  the  sample
are sequestered upon the addition of disodium dihydrogen
ethylenediamine tetraacetate  (Lla-EDTA) .  The  end point of
the reaction is detected by means of Chrome Black  T or
Calmagite, which has a red color in the presence of calcium
and magnesium and a blue color when the cations are
sequestered.

Limitations;

    Range of Applicability;  All concentrations of hardness.

    Interferences;  Excess concentrations  of  heavy metals
    (may be removed by conplexing with cyanide)

Statistical Characteristics;  43 analysts  in  17 laboratories
obtained the following results:

                        Precision      Over Known  Range of
    Accuracy (Bias)     (Std. Dev.)      Hardness as CaCQj

    -0.003 mg/1 (as     2.37 mg/1             31 mg/1
                CaCOj )
    -0.24               2.52                  33
     0.4                4.87                132
    -2.0                2.98                194
    -13.0               9.65                417
    -14.3               3.73                444

Data Outputs;  Not stated.  Assume visual  observation,
manual recording.

Special Sampling Iteguir orients (Collection, Storage, Handling)
Hone.   riaximum holding "time, 7 days.
References;

1.  Methods for Chemical Analysis of Water and VJastes.
    1971.  EPA National Environmental Research Center t
    Cincinnati, Ohio.

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                                47


                        TOTAL HARDNESS


Name of Measurement  Method;   Automated  Method

Principal  Detection  Technique;  Colorinetry

Purpose of Measurenent  (Important Applications);   Surface
waters and saline waters

Summary of Method;   The disodium magnesium EDTA exchanges
magnesium on an equivalent basis for any  calcium  and/or
other cations to form a more  stable EDTA  chelate  than
magnesium.  The free magnesium reacts with calmagite at a pli
of 10 to give a rod-violet complex.  Thus, by measuring only
magnesium concentration in the final reaction stream,  an
accurate measurement of total hardness  is possible.
    Method assumes the use of a Technicon AutoAnalyzer.
Colorimeter is equipped with  520 nru filter.

Limitations:
    Ranges of Applicability;  10 to  400 mg/1 expressed as
    CaC03

Statistical Characteristics:
 Precision;  Results from a single laboratory:

                        Gtd. Dev.      Known Concentration

                        + 1.5 mg/1          19 ing/1 CaCO..
                        + 1.5              120          ^
                        + 4.5              385
                        + 5.0              366

 Time of Measurement;
 12 determinations per hour.  30 minutes warm up.

Calibration Requirements;  Apparatus must be calibrated against
stock solutions of known concentration.

Data Outputs;  Analog electrical signal, recorded on stripchart,

References;

1.   Ilethods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.   Technicon AutoAnalyzer Methodology.  1960.  Bulletin No. 2,
    Channcey, New York,

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                               48
                           METALS

Name of Measurement Method;  Atomic Absorption

Parameter(s) Measured;  Metals

                        Aluminum    Copper       Potassium
                        Arsenic     Iron         Silver
                        Cadmium     Lead         Sodium
                        Calcium     Magnesium    Zinc
                        Chromium    Manganese

Principal Detection Technique;  Atomic Absorption
Spectrescopy.

Purpose of Measurement (Important Applications);  Ra^id
determination of certain metals in ambient surface  waters,
domestic and industrial water, and saline  waters.

Summary of Method;  Atonic absorption o^ectroocopy is
similar to flame emission photometry in that a sample is
atomized and aspirated into a flame.  Flame photometry,
however, measures the amount of light emitted, whereas, in
atomic absorption spectrophotometry, a light beam is
directed through the flame into a monochromator, and onto a
detector that measures the amount of light absorbed.  In
many instances absorption is more sensitive because it
depends upon the presence of free, unexcited atoms and
generally the ratio of unexcited to excited atoms at a given
moment is very high.  Since the wavelength of the light beam
is characteristic of only the metal being determined, the
light energy absorbed by the flame is a measure of the
concentration of that metal in the sample.  This principle
is the basis of atomic absorption spectroscopy.

Analytical procedures specific to each of the metals listed
above are given in the Reference cited.

Limitations:
    Range of Applicability;  Detection limits,
    sensitivities, and optimum ranges of concentrations,
    vary with make and model of atomic absorption
    spectrometer.  The following table provides some
    indication of ranges of measurement.  In many cases the
    range can be extended higher or lower by instrument
    adjustment, use of a different wavelength, or sample
    pretreatment.

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.Metal
Aluninui.i
/vrsenic
Cadmiuu
Calciun
C'nroiniun
Copper
Iron
Lead
"lagnesiun
'langaneso
Potassium
Silver
Sodium
Zinc
              Detection
                Li nit
              (IKJ/1)

                 0,1
                 0. 05
                 0.001
                 0.003
                 0. 01
                 0.005
                 0.004
                 0.01
                 0.0005
                 0.005
                 0.005
                 0.01
                 0.001
                 0. 005
r>enr;itivi ty
  (no/D

   0.4
   1.0
   0.00-1
   0.07
   0.02
   0.04
   0.006
   0.06
   0.005
   0.04
   0.01
   0.05
   0.003
   0.02
G.-.>tii..ui.i Concentration
        rinnge
         (nuj/1)
    Accuracy  anc1 Precision Data:
                         Hctal
'lotal               Concentration
                         (ug/1)

Direct  Jetornination
Cadriiun
Chroniuu
Co-ver
Iron
May no si urn
.'lanijanese
Silver
Zinc
50
50
1000
300
200
50
50
500
                                     Rolativo
                                      Error
                                     (Percant)
                                         3.15
                                         2.29
                                         3.42
                                         0.6-1
                                         £.30
                                         6.00
                                        10.57
                                         0.41
10
10
0.1
1
1
0.1
0.1
1
0.01 -
0.1
0.01 -
0.1 -
1
0.1
1000
100
2
200
200
10
20
10
2
20
2
20
200
2
                          Relative
                   Standard Deviation
                         (Percent)
                                                          21.62
                                                          26. 44
                                                          11.23
                                                          16.53
                                                          10.49
                                                          13.50
                                                          17.47
                                                           3.15
Extracted Gannlos
Cr.dniurn
Load
                          10
                          50
         3.03
        ID.00
        72.77
        23.46

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                              50
    Time of Measurements  Rapid

Calibration Requirononts;  See Reference

Data Outputs;  Electrical signals

Special Sampling Requirements  (Collection! Storage, Handling)
See Reference

References:
1.  iletliods for Chemical Analysis of Water and Wastes.
    1971.  EPA national Environmental Research Center,
    Cincinnati, Ohio.

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                               51


                          MERCURY

Marae o£ Measurement. Method;  Flaraeless Atomic Absorption

Principal Detection Technique;  Atonic Absorption
Spectroscopy

Purpose of Measurement  (Important Applications);  Ambient
surface waters, saline waters, wastewaters, and effluents.
Hay also be used for fish tissue, mud, sediments, and other
materials following proper digestion.

Summary of Method;  The flameless AA procedure  is a physical
method based on the absorption of radiation at  253.7 nm by
mercury vapor.  The mercury is reduced to the elemental
state and aerated from solution in a closed system.  The
mercury vapor passes through a cell positioned  in the light
path of an atomic absorption spectrophotometer.  Absorbance
(peak height) is measured as a function of mercury
concentration and recorded on a stripchart.

Limitations:
    Range of Applicability;  Detection limit is 0.2 ug/1
    mercury

    Interferences;  Sulfides, copper, high concentration of
    chlorides, and certain volatile organics

Statistical Characteristics;  Using an Ohio River composite
sample with a background mercury concentration of 0.35 ug/1,
spiked with concentrations of 1, 3, and 4 ug/1, the standard
deviations were ±0.14, +0.10 and +0.03 ug/1 respectively.
Standard deviation at tKe 0.35 level was 0.16.  Percent
recoveries at the three levels were 89, 87, and 87%
respectively.

Calibration Requirements;  Detailed calibration instructions
i
meluued in Reference below.

Data Outputs;  Analog electrical signal, displayed on
stripchart.

Special Sampling Requirements  (Collection, Storage, Handling)
Acidify sample to pll of 2 or lower.

Reference;

1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

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                              52
                          MERCURY

Name of Measurement Method;  Cold Vapor Technique -
 (Biological Materials)

Medium;  Fish tissue  (and other biological materials)

Principal Detection Technique;  Atonic absorption
spectroscopy

Purpose of Measurement  (Important Applications); To measure
total mercury(organic  and inorganic)in  fish and in other
biological materials.

Summary of Method;  Weighed portion of sample is digested
with sulfuric and nitric acid and oxidized overnight with
potassium permanganate.  Mercury content  is measured by  the
conventional cold vapor  (Flaineless AA) technique summarized
in Method Summary for Mercury in Water  (this compendium) and
described in References  (1) and  (2) of that Summary.

Limitations:
    Range of Applicability;  0.2 to 5 ug/g.   (May be
    extended by varying sample size or through instrument
    control).

    Interferences;  Most interferences are destroyed during
    digestion and oxidation steps.

    Pitfalls; Special Precautions;  See "Sampling
    Requi renents" below.

Statistical Characteristics:
    Precision;  The following standard deviations on
    replicate fish samples were recorded at the indicated
    levels:  0.19 ug/g+0.01; 0.74 ug/g+0.05; and 2.1
    ug/g+0.07.  The coefficients of variation at these
    leveTs were 11.9 percent, 7 percent, and 3.6 percent
    respectively.

    Time of Measurement;  Several hours.

Data Outputs;  Electrical signals displayed or recorded,

Special Sampling Requirements (Collection, Storage, Handling):
Mercury is not uniformly distributed throughout the whole
fish and it is therefore necessary to decide in advance which
part of the fish is to bo analyzed.  In the case of a large

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                             53
specimen, only a selected part of the fish may be examined.
(Ordinarily, only the edible flesh is analyzed).  In any
event, the portion analyzed should be reported.

If it becomes necessary to freeze the fish before analysis,
the sample should not be allowed to thaw before weighing as
higher results may be observed.

References:
    Mercury in Fish.  1972.  Method Description received
    from Analytical Quality Control Laboratory, EPA
    national Environmental Research Center, Cincinnati,
    Ohio.

    lithe, J.F, , F.A.J. Armstrong and M.P. Stain ton*
    Mercury Determination in Fish Samples by Wet
    Diyestion and Flameless Atomic Absoprtion
    Spectre-photometry.  Jour. Fisheries Research
    Board of Canada, 27:805.

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                                 54
                          MERCURY

Name of Measurement Method;  Gas Chronatography  (In Fish)

Medium;  Fish Tissue

Purpose of Measurement  (Important Applications); To
determine concentration of mo thy 1 mercury in fish samples of
all typos.

Gur.unary of Method; ' A measured weight of fish is treated
with acid and bromide salt.  Methyl mercury is extracted as
methyl mercury bromide with toluene.  A cleanup operation is
performed by first extracting the toluene layer with aqueous
solution of sodium thiosulfate.  The methyl mercury
thiosulfate complex formed in the aqueous layer is then
reacted with an excess of potassium iodide.  Denzone is used
to extract any methyl mercury as the iodide salt.  A portion
of the benzene extract is chromatographed directly and
response is compared to standard responses.  The iodide is
detected using gas chromatography with an electron capture
detector.

Limitations:
    Sensitivity;  0.01 ug/g

    Interferences;  Materials in sample will generally not
    cause a problem.  Toluene from extraction not removed by
    cleanup step may interfere.

    Pitfalls; Special Precautions;  Potassium iodide
    solution decomposes readily.Any free iodine may cause
    interfering peaks on chromatogram.

Statistical Characteristics:
    /vccuracy and Precision;  When seven portions of a filet
    of a large white perch were analyzed by this method, a
    mean value of 0.37 ug/g was obtained, with a standard
    deviation of 0.034 ug/g.

    Recoveries of methyl mercury chloride injected into this
    same fish at the 0.20 ug/g level averaged 95.5 percent.

    Time of Measurement;  Approximately one hour.

Calibration Requirements;  No unusual requirements.
Calibration procedure given in Reference.

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                                  55
Data Outputs;  Analog electrical signal recorded as
chromatogram,

Special Sampling Requirements  (Collection, Storage/ Handling);
Analysis should be performed oit the fish before decay has begun.
Fresh or frozen fish can be analyzed with confidence; results
on partially decomposed samples must be viewed with caution and
reported as such.

Fish samples can dehydrate rapidly unless protected during
handling.  Defrosting and refreezing a frozen sample before
analysis, and permitting a sample to stand open at room
temperature before weighing must be avoided.

Selection of portions of the fish must be of a consistent
nature and properly identified as to location on the specimen.

References:
1.  Methyl flercury in Pish.  1972.  Provision Method
    Description received from Analytical Quality Control
    Laboratory, EPA National Environmental Research
    Center, Cincinnati, Ohio.

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                                 56
                          MERCURY

Medium;  Sediment

Name of Measurement Method:  Gas Chromatography  (In
Sediment)

Principal Detection Technique;  Electron capture gas
chromatography

Purpose of Measurement  (Important Applications).;  This
method is applicable to bottom samples such as mud, sludge,
silt and gravel.

Summary of Method;  A measured weight of sediment is treated
with acid and bromide salt.  Methyl mercury is extracted
with toluene as methyl mercury bromide.  A cleanup operation
is performed by first extracting the toluene layer with an
aqueous solution of sodium thiosulfate.  Thu methyl mercury
thiosulfate complex formed in the aqueous layer is then
reacted with an excess of potassium iodide.  Benzene is used
to extract any methyl mercury as the iodide salt.  A portion
of the benzene extract is chromatographed directly and
response is compared to standard responses.  The iodide is
detected using gas chromatography with an electron capture
detector.

Limitations;

    Sensitivity;  0.001 ug/g

    Interferences;  Materials in sample generally will not
    cause a problem*

    Pitfalls; Special Precautions;  Potassium iodide
    solution may decompose to free iodine,  which may cause
    interfering peaks on the chromatogram.

Statistical Characteristics;

    Accuracy and Precision;  Ten dfferent sediment samples
    with a background level of less than 0.0005 ug/g were
    spiked with 0.010 ug/g and measured with this teat.  The
    average was 0.00963 ug/g and the standard deviation was
    0.00087 ug/g.

Calibration Requirements;  Given in Reference below

Data Outputs; Analog electrical signal recorded as
chrornatogram.

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                               57

Special Sampling Requirement  (Collection, Storage, Handling);
It should be understood that methyl mercury is generated
froia inorganic mercury by biological methylation.  Since
this can occur at widely varying rates in samples stored in
a laboratory, analyses intented to determine the methyl
mercury concentration existing at the sampling site at the
time of sampling should be performed as quickly as possible.
Storing samples frozen or refrigerated before analysis is
acceptable but is a less desirable alternative.

References;

1.  Ilethyl Mercury in Sediment.  1972.  Provision Method.
    Description received from Analytical Quality Control
    Laboratory, EPA national Environmental Research Center,
    Cincinnati, Ohio.

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                               58


               ATOMIC ABSORPTION SPECTROSCOPY

IJame of Measurement Method;  Mercury — Cold Vapor Technique

Purpose of .Measurement  (Important Applications);  This
metnod is applicable to surface waters,saline waters,
wastewaters, effluent and domestic sewage.

S ummary of Metho d;  The atonic absorption procedure is a
physical method based on the absorption of radiation at
253.7 nrn by mercury vapor.  The mercury is reduced to the
elemental state and aerated from solution in a closed
system.  The mercury vapor passes through a cell positioned
in the light path of an atomic absorption spectrophotoraeter.
(Instruments designed specifically for the measurement of
mercury using the cold vapor technique are commercially
available and may be substituted for the atomic absorption
spectrophotometer).  Absorbance  (peak height) is measured as
a function of mercury concentration and recorded in the
usual manner.

In addition, inorganic forms of mercury, organic mercurials
may also be present in an effluent or surface water sample.
The organomercury compounds will not respond to the
flameless atomic absorption techniques unless they are first
broken down and converted to mercuric ions.  Potassium
permaganate oxidizes many of these compounds but recent
studies have shown that a number of organic mercurials,
including phenyl mercuric acetate and methyl mercuric
chloride, are only partially oxidized by this reagent.
Potassuim persulfate has been found to give approximately
100 percent recovery when used as the oxidant with these
compounds.  Therefore, a persulfate oxidation step following
the addition of the permanganate has been included to insure
that organomercury compounds, if present, will be oxidized
to the mercuric ion before measurement.  A heat step is
required for methyl mercuric chloride when present in or
spiked to a natural system.  For distilled water the heat
step is not necessary.

Limitations;

    Range of Applicability;  The range of the method may be
    varied through instrument and/or recorder expansion.
    Using a 100 ml sample, a detection limit of 0.2 ug Hg/1
    can be achieved; concentrations below this level should
    be reported as 0.2.

    Interferences:  Sulfides  (interference can be eliminated
    by addition of potassium permaganate), high
    concentrations of copper and chlorides.   (Certain
    volatile organic materials may also interfere.)

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                                59
Statistical Characteristics :

    Accuracy and Precision;  Usiny  an Ohio  River  composite
    sample with a background mercury concentration  of  0.35
    ug/1, spiked with concentrations of  1,  3  and  4  ug/1,  the
    standard deviations were +0.14, +_0.10 and +0.03 u
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                               60


          METIIYLEnr BLUE ACTIVE SUBSTA-JCLS  (M1JAS)


Uarao of Measurement .'lotliod;  "lethyleno Blue "lethod

Principal Detection Technique;  Colorinetry

Purpose of Measurement  (Important Applications):
Determination of concentration of detergents, phosphates,
surfactants in drinking water, ambient surface waters,
domestic and industrial wastes.

Summary of I lethod:  The dye, muthylene blue, in aqueous
solution reacts with anionic-type surface active materials
to form a blue colored salt.  The salb is extractable with
chloroform and the intensity of color produced is
proportional to the concentration of IIHAS.

Limitations;

    Range of Applicability;  0.025 to 100 my/1 of M3AS
    (expressed as linear alkyl sulfonate - LAS)

    Interferences;  Chloride at concentrations over  1,000
    rug/1 sulfonates, carboxylates, phosphates, phenol,
    cyanates, and thiocynaides (at concentrations higher
    than normally encountered in water or wastev/ater).

Statistical Characteristics;

    Accuracy;   +1.2% to -11% bias  (conditions as given for
    Precision).

    Precision;  Determinations by 110 analysts exhibited
    relative standard deviations of 10 to 15 percent  for
    known concentrations of :1BAS ranging from 0.27 to 2.94
    mg/1 of LAS

Data Outputs;  Analog electrical signal, displayed on ranter.

References;  Methods for Chemical Analysis of Water  and Wastes,
             EPA  National  Environmental Research Center,
             Cincinnati, Ohio  (1971).

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                                 61
                          NITROGEN



Name of Measurement Method;  Ammonia Distillation Procedure

Principal Dectection Technique;  Colorimetry
   pose of Measurement  (Important Applications);  Ambient
   face waters/ domestic and industrial wastewate.rs,  and
Pur
sur
saline waters.
Summary of Method;  The sample is buffered at pH of 9.5 x*ith
a borate buffer in order to decrease hydrolysis of cyanatea
and organic nitrogen compounds, and is then distilled into a
solution of boric acid.  The ammonia in the distillate can
be determined either colorimetrically by nesslerization, or
titrimetrically with standard sulfuric acid with the use of
a mixed indicator, the choice between these two procedures
depending on the concentration of the ammonia.

Limitations;

    Range of Applicability;  0.05 to 1.0 mg/1 ammoniacal
    nitrogen

    Interferences;  Cyanates, certain alcohols, aldehydes,
    and ketones may interfere.  (Certain amines would
    interfere if distillation step were not included.

    Pitfalls; Special Precautions;  Residual chlorine must
    be removed.  If sample is preserved by a mercury salt,
    the mercury must be complexed.

Statistical Characteristics;  24 analysts in 16 laboratories
obtained;

Known Concentration    Accuracy (Bias)       Precision
in mg/1 Nitrogen       mg/1 N                (Std. Dev.) mg/1

     0.21                 -0.01              0.122
     0.26                 -0.05              0.070
     1.71                 -1-0.01              0.244
     1.92                 -0.04              0.279

Calibration Requirements;  Set up a series of standards in
Nessler tubes.  Care needed in calibrating for saline water
tests.

Data Outputs;  Meter reading (analog electrical signal)

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                                62
SjGcial Sampling requirements  (Collection.,  Storage/  Handling)
Preserve with mercuric chloride ami storo at4* C."
1.  'totiiods for Chemical Analysis of Water  and  Y7astQS.
    1971.  ]-]PA National Environmental Research  Center/
    Cincinnati/ Ohio.

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                                63
                          NITROGEN
Uaiae of Measurement Method;  Ammonia

Principal Detection Technique;  Titrimetric

Purpose of Measurement  (Important Applications):  Surface
waters, domestic and industrial wastewaters, and saline
waters.

Sunimajry of Sethodr  .The" saznpl&risUauf fered at pH of 9 ..5. .wi±h«
Derate buffer in order-to decrease hydrolysis- of cynates and
organic nitrogen ammonia in the^distillate can be determined
either coloiTOtrically- by hesslerlzation or titrintrically
with*standard/sulf uric~acid and. the use of a mixed-ihdicator,
the*choice between-these two procedures'depending on the
concentration of the ammonia.

Limitations:

    Range of Applicability;  1.0 to 25 mg/1 ammoniacal
    nitrogen.

    Interferences;  Cyanatas, hydrozine and similar
    compounds.

    Pitfalls; Special Precautions:  Residual chlorine and
    mercury must be removed.

Sampling Sampling Requirements (Collection/ Storage/ Handling)
Preserve with mercuric chloride and store at 4"C.

References;

1.  Methods for Cnemicai Analysis of Water and Wastes.
    1971.   EPA National Environmental Research Center,
    Cincinnati, Ohio.'

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                                    64
                               NITROOFN


Name of "oasurement -Method;  Ammonia

Principal Detection Method;  Automatic Colorimetric

Purpose of Measurement  (Important Applications);  Surface Waters,
salinewaters.

Summary of Method;  The intensity of the indophenol blue color, formed
by the reaction of ammonia with alkaline phenol hypochlorite, is
measured.  Sodium nitroprusside is used to intensify the blue color.

    Uses Technicon AutoAnalyzer.  Colorimeter is ecmipped with  360 or
    650 filters.

Limitation;

    Range of Applicability;  .01 to 2.0 mq/1 nitrogen as NFL,

    Pitfalls; Special Precautions;  Harked variations in pH among
    samples should be eliminated.

Statistical Characteristics;

    Precision;  In a single laboratory using surface water at water at
    ~~         concentrations of 1.41, 0.77, 0.59, 0.43 mg/1 of  NIU-N,
              the standard deviation was £.005 rag/1.             ~

Calihcation Requirements;  Requires calibration against samples, both
for fresh water and substitute ocean water of specified constituency.

Data Outputs;  Analog electrical signal displayed on stripchart

Special Sampling Requirements  (Collection, ftorage, Handling);
Preserve with mercuric chloride and refrigerate at 40*Max. holding
tine - 7 days

References;

1.  Methods for Chemical Analysis of Water and Wastes.  1971.   EPA
    national Environmental Research Center, Cincinnati, Ohio.

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                                   65
                               NITROGEN


Name of Measurement Method;  Kjeldahl

Principal Detection Technique:   Tritriraetric

Purpose of_JMeasurement  (Important Applications): .  Stir face waters,
domestic and industrial wastes,saline waters.

Summary of Method;  Total Kjeldahl nitrogen  is defined as the  sum  of
the free ammonia and organic nitrogen compounds which are converted to
ammonium sulfate under defined conditions of digestion.  The procedure
converts nitrogen components of  biological orgin such as amino acids,
peptides, and proteins to ammonia, but may not convert the nitrogenous
compounds of certain industrial  wastes such  as amines, hydrazones,
nitro compounds, and others.

    The sample is heated in the  presence of  concentrated sulfuric
acid, Itj SO./)  and HgSfy and evaporated until S0«j fumes are obtained
and the solution becomes colorless or pale yellow.  The residue  is
collected, diluted, and is treated and made  alkaline with a hydroxide-
thiosulfate solution.  The ammonia is distilled and determined after
distillation either by nesslerization or titrimetrically.

Limitations;

    Range of Applicability!  Concentration above 1 mg/1 of nitrogen.

Statistical Characteristics;  Results obtained by  31 analysts  in 20
laboratories

Known Cone, of     Precision as          Accuracy as
Nitrogen, Kjeldahl   Standard Deviation    Bias,       Bias,
mg N/liter	mg^N/lrter            mg N/liter

                                                      + .03
                                                      + .02
                                                      + .04
                                                      -.08

Special Sampling Requirements (Collection, Storage, Handling);
Unstable.  Analyze soon after sample is acquired.

References;

1.  Methods for Chemical Analysis of V7ater and Wastes.  1971.    EPA
    National Environmental Research Center, Cincinnati, Ohio.
0.20
0.31
4.10
4.61
0.197
0.247
1.056
1.191
+15.54
+ 5.45
+ 1.03
- 1.67

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                                    66
                               NITROOFH


Name of Measurement Method;  Kjeldahl

Principal Detection Technique:  Automated Phenolate

Purpose of Measurement  (Important Applications) ;  Ambient  surface
waters, saline waters, donest.i.c ana industrial  wa s t e s .

Summary of Method ;  The sample is autonatically digested with a
sulfuric acid solution containing potass.ium  sulfate and mercuric as  a
catalyst to convert organic nitrogen to ammonium sulfate.   The
solution is then automatically neutralized with sodium hydroxide
solution and treated with alkaline phenol reagent and sodium hypo-
chlorite reagent.  This treatment forms a blue  color desiqnated as
indophenol.  Sodium nitroprusside, v;hich increases the intensity of
the color, is added to obtain necessary sensitivity for measurement  of
low level nitrogen.

Utilizes Technicon AutoAnalyzer.  A colorimeter with 630 nm filter is
used.

Limitations:

    Range of Applicability;  Nitrogen concentrations from  0.05 to 2.0
    Interferences ;  Iron, chromium, or copper ions may interfere

Statistical Characteristics!  Results from six laboratories analyzing
natural waters with known concentrations of Kjeldahl nitrogen:
Known' "Conc. of       Precision as                  Accuracy as
Kjeldahl-Nitrogen    Standard Deviation       Bias         Rir»s
mg N/liter _ Kjeldahl-N, mg N/liter     %      ma N/liter
1.89
2.18
5.09
5.81
0.54
0.61
1.25
1.85
-24.6
-20.3
-23.0
-21.9
- .46
- .62
-1.21
-1.27
Time of Measurement;  About 20 samples per hour

Calibration Requirement;  Requires preparation of standardizations

Data Outputs;  Analog electrical signal recorded on stripchart

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                                   67
Special Saraplinq_Requirenents  (collection, storage/ handling);
Preserve with mercuric chloride; refrigerate at 4'C.Analyze
immediately after acquistion.

References;

1.  Methods for Chemical Analysis of V7ator and TTantos.   1971.  EPA
    national Environnental Research Center, Cincinnati,  Ohio.

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                                   68
ITane of "easurcnent Method;   Kjeld.ahl

Principal ^Detection Technique ;   Colori'-'etrir

Purpose of Measurencnt (. Important Applications) ;   Anhient surface
T 7.1. tors, saline waters, domestic anr! industrial  v;a ste s .
      ^ of "othocl;   Heo r-unnar" for ITitrorrnn,  I'jnlc'ahl,  r'
Color ir>e trie.   Tor  ?Te.nsleriz;ation, a spectronhonoter  or  color inetcr in
upod, filte>-orl  at 400  to 525 nn.
Linitations;

    Range of Applicability;   ConcGntrationn of  nitroqon helov 1 ma/1
                 Characteristics;  Sep Tethocl  Surnmary for nitrogen,
    Kjolc^ahl,  "otal  (Ti trine trie)

Data  utputs;   "eto.r  roarling (analon olectrical  sianal), recorded


                  ^eauirenonts (Collection, ftoracro,
Una tab In , analyse  iTneylip.telv after sanplc is acquired.

References;
1.  ;:ethorir>  for  Chemical Analysis of T^ter and barter;.   1971.  FP71
    national HnvironnRntal Research Center, Cincinnati,  Ohio.

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                               69
Principal Detection Technique;   Nitrite

Purpose of Measurement  (Important  Applications) ;   Anb.ient surface
waters, saline waters,  domestic  ami  industrial wa s te s .

Summary of Method ;  The diazoniun  cornound  forned  by dinzotation of
sul'f anITanide by nitrite  in water  under  acid  conditions is coupled
with IT- (l-naphtyl)-ethylenedianine to  produce a reddish -purple color
which is read in a snectro photometer at  540nn against a blanl:.
Concentration of lio^-n  is plotted  aqainst optical  density.

Limitations;

    Range of Applicability;   0.05  to 1.0 mq/1 nitrite nitrogen

    Interferences ;  Strong oxidizing or  reducing aqents or hirrh
    alkalinity


Calibration Requirements ;  Requires  use  of  standard solutions

Data Outputs;  lieter reading  (analorr
Special Sampling Requirements  (collection ,  Storage, Handling) ;
Preserve with mercuric chloride; refrigerate  at 4feC.   "axinun storage
tine - 7 days.

References;

1.  Methods of Chemical Analysis of Uater and Wastes.   1971.   EPA
    National Environmental Research Center, Cincinnati,  Ohio.

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                              70
Mane of "casurernnt Method;  nitrate-nitrite

Principal Detection Technique;  Automated  Cadmium Deduction T'ethod

Purpose of ileasureiuent  (Important  Applications);   Determines nitrates
and nitriter;,n.i.n<7lv or combined,  in  surfaco.  and  saline waters.

nunnary of Method;  The initj.nl ste^  ir?  to reduce the  nitrates to
nitritor; b^ using a cadniun-cop'inr catalyst.   The nitrites (thosr
orrrinallv present plus roducerl nitratos) are  tho.n reacted v:ith
snlfanilamdo to forn the c!iazo compound v-'l^ich in thon coupled in an
acid solution  (p!1 2.0-2.5) with N-l nanhtylethylene^iamine
hydrochlorido to forri the azo dye.  The  azo values are readily
obtainable by carryina out the procedure—first with,  and then
without, the initial Cd-Cu reduction  step.

A Technicon AutoAnalyzer is  used.   The colorireter is  equipped with
540 nn filters.

Limitations;

    Range of Applicability;  0.05  to  10.0  mg/1 nitrogen present as
    nitrate.             *"

    Interferences;  Armenia  and primary  anines; some nctal ions
    (mercury and copper )  nay produce interfering color complexes.

Statistical Charatoristics;  Three laboratories analyzed four natural
water samples containing exact concentrations of  inorganic nitrate,
with the following results:
Known Cone, of     Precision  as               Accuracy as
nitrate nitrogen    Standard  Deviation     Bias,           Bias,
mg N/liter	mg IJ/liter	%	mg N/liter

    0.29                  0.012              + 5.75     +.017
    0.35                  0.092              +18.10     +.063
    2.31                  0.313              + 4.47     +.103
    2.48	0.176	- 2.69     -.067	

Time of Hoasurenent;  Not stated,  but assumed to be rapid (i.e.,
several samples per hour.

Calibration^Reguireinonts:   Calibration  against standard solution is
necessary."Analysis of  saline waters requires calibration against
substitute ocean water.

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                                  71
Data Outputs:  Analoa electrical  signal disnlayml on stri^chart.
 ne     fanplincr ^eguirnnonts  (collection ,  storage, handling); S
in not ntnble since anin^  in  natural v;aterc. nay rnr.ct v.rth nitrites.
     n shonlrl IP annl^^er1 an soon  an possible after acnnqition.
References:
1.  netliotiR for Chonical  Annlvsis o^ T1ator nrv1 fantos.  1971.  EPA
    national Environmental  Research Con tor, Cincinnati, Ohio.

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                               72


                               NITROGEN


Name of Measurement Methodt  Nitrate and Nitrite

Principal Detection Technique;  Automated Hydrazine Reduction Method

pvirpose of Measurement__(Important Applications) t  Ambient  surface
waters and domestic or industrial wastes.

Summary of Method;  This method, using the Technicon AutoAnalyzer,
determines NO«-N by the conventional diazotization-coupling reaction.
The NQj-N is reduced with hydrazine sul^ate in  another portion of the
sample and the. nitrite thus formed is determined in the usxial manner.

    Subtraction of the NQ^-N orginally present  in the sample from the
    total NCU-N will give the orginal NOa-N concentration  in terms  of
    NO^-N.  -        "                  *

    Utilizes Technicon AutoAnalyzer.

Limitations;

    Range of Applicability;  Nitrite or nitrate nitrogen in
    concentrations of 0.05 to 10.0 ing/liter.

    Interferences;  The following substances may interfere
    (concentration below which interference will not occur are listed
    in Reference);  chlorides, phosphates, sulfides, ammonical
    nitrogen, manganese, calcium, and ferric ions, and ABP.

    Pitfalls; Special Precautions;  Toxic reagents

Statistical Characteristics;  Nine laboratories analyzed four natural
water sample's containing exact concentrations of inorganic nitrate,
with the following results:
Known Cone, of     Precision as                Accuracy as
Nitrate Nitrogen    Standard Deviation    Bias,         Bias,
mg N/liter	mg N/liter	%    	mg N/liter
0.29
0.35
2.31
2.48
0.053
0.050
0.250
0.217
-0.8
+1.9
+3.0
-1.2
.002
.007
.07
.03
In a single laboratory, using surface water samples at concentrations
of 0.1, 0.2, 0.8, and 2.1 mg-N/1, the standard deviations were +0.04,
+0.05, and +0.05, respectively.

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                                73
Time of Measurement;  About 20 samples per hour

Calibration Requirements;  See Reference.

Data Outputs;  Analog electrical  sirrnal displayed  on  stripchart.

Special Sampling Requirements  (collection, storage/ handling);
Preserve sample with mercuric chloride.  Pefriqernte  at  4*?C.  Haxinum
holding tine - 7 days.

References;

1.  .Methods for Chenical Analysis of T-Tater and TJastes.   1971.
    National Environmental Research Center, Cincinnati,  Ohio.

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                                 74
                               NITROGEN


Name of Measurement Method;  Organic Plus Ammonia

Principal Detection Technique;  Automated Phenolate Method

Purpose of Measurement  (Important Application):  Ambient  surface
waters and saline  waters.

Summary of Method;  Organic nitrogen is determined by manually
digesting the sample with potassium persulfate and sulfuric acid to
convert the organic nitrogen, and any ammonia present, to ammonium
sulfate.  Subsequently, the automated phenol-hypochlorite procedure  is
used to measure the ammonia nitrogen.  Nitrate-nitrite nitrogen is not
measured by this procedure.

Utilized Technicon AutoAnal^'zer with colorimeter eauinped with 650 nr>
filter.

Limitations;

    Range of Applicability;  Nitrogen concentrations of 1.0 to 10.0
    mg/1                 ""

    Time of Measurement;  Not stable.  Estimated to be between one and
    two hours per determination.

    Calibration Requirements;  Calibrate against standard solutions

Data Outputs;  Analog electrical signal recorded on stripchart.

Special Sampling Requirements (collection,storage,handling);
Preserve with mercuric chloride.  Refrigerate at 4*jC.  Maximum storago
time - 7 days.

Reference;

1.  Methods for Chemical Analysis of Water and TTantes.  1971.  EPA
    National Environmental Research Center, Cincinnati, Ohio.

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                                  75
                                             ACID
Name of T'oasureraent^ethod;   Zinc-Zincon  .Method

Principal Detection Technique;  Colorinetry

Purpose of Measurement  (Important Applications)   Ambiont surface
waters  (non-saline) .

Summary _of Method ;  Zinc forms a blue-colored complex  vith  2  carboxy-
2 • -hydroxy^S f -sul f o f ormazy Ibenzone  (Zincon)  in  a  solution buffered  to
pll 9.2.  When IITA is added,  the Zn-Zincon  complex is broken vhich
reduces the optical density  in proportion  of the  anount of  IITA
present.

A photoneter enuipped with 620 nn filter is  used.

Limitations;

    Range of Applicability;   0.5 to  10.0 mg/1 1TTA

    Interferences ;  Cations  of conrcon netals (calciun, naqnesium,
    zinc, copper iron, nanganesp, et al) .

    Pitfalls; Special Precautions;   Fost interfering cations  can be
    renoved by ion-exchanae  resin.

Statistical Characteristics;

    Precision;  In a single  laboratory,  using spihed surface  water
    samples at concentrations of 0.5/2, 6,  and 10 mg/1 NTA,  standard
    deviations were 4-0.17, 4-0.14, +0.1,  and  4-0.16, respectively.

Calibration Requirements;  Require stock solution

DataniOutj3uts;  Analog electrical signal  displayed on meter.

Special Sampling Requirements  (collection  storage , handling) ;  Sample
should Ee analyzed as soon as possible after acquistion,  since IITA  is
biodegradable .

References;

1.  Ilethods for Chemical Analysis of Uater and  Wastes.   1971.   EPA
    National Environmental Research  Center,  Cincinnati,  Ohio.

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                                 76



                            _jTjrr-,^TjJnTni.7V.crTic  AC in  (IITA)


Tlane of r.easurement Method;  Automated  Zinc-Zincon J'ethod

Principal Detection Technique;  Color ipo try

Purpose of "easurenent  (Important Applications) ;   Ambient surface
v/atorn  (non-nnline) .

Summary of "ethod;  Zinc forms a blue-colored  corplcr with 2-carboxv-
2 ' -hvdroxy-5 ' sulfof ornazylbFinzene  (7incon)  in  a  solution buffered to
nil 9.2.  Uhen TITA is added, the Zn-Zincon conplex if? broken which
reduces the ontical density in proportion to the anount of ITT7\
present.

A Tehonicon AutoAnaly^er is used.  The  colorineter irs equipped with
600 to 625 nn filter.

    Limitations!
          of Applicability;  0.0^  to  l.o mn/i  Or 0.5  to 10.0 mq/1 TITA,
    denendinq on tyr»c of nanifold  sy
    Interferences;  Common cations  (calcium,  naqnesinm,  copper iron,
    nangaqnese) .  Constituents of sewaqe  cause  sor.e interference also.

    Pitfalls; Special rrecautions;  Not applicable to saline water.

Statistical Characteristics;

Reproducihility;  In a single laboratory,  using surface  water san.ples
at concentrations .of 0.1, 0.10, 0.27,  and 0.44  ma/1,  the standard
deviations v?ere +0.01, +0.004, +0.005, respectively.   At
concentrations of 1.3, T. 0,  5.8, and  7.4  nq/1,  the standard deviations
v;ere +0.05, +0.05, +0.07, and +0.1, respectively.

Time of Measurement;  About  13 samples per hour.

Calibration Requirements ;  Requires standard  solutions.

Data Outputs ;  Analog electric signal, recorded automatically.

Special Sampling Requirements Collection,  Ptorage, Tandling) ; Analyze
soon after sample collection because  of biodeqradability of HTA .

Reference;

1.  Methods for Cheraical Analysis of  Water and  Wastes.   1971.  EPA
    National Environmental Research Center, Cincinnati,  Ohio.

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                               77


                              PHOSPHORUS


Name of Measurement .Method;  Single Reagent Method

Principal Detection Technique;  Colorimetry

Purpose of Measurement  (Important Applications);  Ambient surface
waters, drinking water, domestic and industrial wastes, saline  waters
(may also be applicable to sediments, sludges, and algae blooms).

Summary of Method;  Ammonium molybdate and potassium antimonyl-
tartraie react in an acid medium with dilute solutions of phosphorus
to form an antimony-phospho-molybdate complex.  This complex is
reduced to an intensely blue-colored complex by ascorbic acid.  This
color is proportional to the phosphorus concentration.

Only orthophosphate forms a blue color in this test.  Polyphosphates
(and sone organic-phosphorus compounds) may be converted to the
orthophosphate form by sulfuric-acid-hyc'.rolysis.  Organic phosphorus
compounds may be converted to the orthophosphate form by sulfuric acid
hydrolysis.  Organic phosphorus compounds may be converted to the
orthophosphate form by persulfate digestion.

A spectrophotometer of filter photometer suitable for measurements at
880 nm is used.

Limitations;

    Range of Applicability;  0.01 to 0.5 mg/1 phosphorus

    Interferences;  High concentrations of iron.  Also, mercuric
    chloride (when used as a preservative).

    Pitfalls; Special Precuations;  Avoid use of commercial detergents
    for cleaning analyzed natural water samples containing exact
    concentrations of organic phosphate, with the following results;
Known Cone, or    Precision as                    Accuracy as
Total Phosphorus    Standard Deviation    Bias,       Bias,
mg P/liter	mg P/liter             %        mg P/liter
0.110
0.132
0.772
0.882
0.033
0.051
0.130
0.128
+ 3.091
+11.99
+ 2.96
- 0.92
+.003
+ .016
+ .023
-.008

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                                 78
Twenty-six analysts in sixteen laboratories  analyzed  natural  water
samples containinq exact concentrations of orthophosphatc,  with the
following results:
Known Concen. ofPrecision an             Accuracy  as
Orthophocphate      Standard Deviation    Bias,        Bias,
nq P/liter          nq P/liter      	%	ng P/liter
0.020
0.038
0.335
0.383
0.010
0.003
O.Olfl
0.023
-4.05
-P. 00
-2.75
-1.76
-.001
-.002
-.000
-.007
Calibration Requirements;  Requires standard  solutions

Data Outputs;  Teeter  (analoa voltaqe), nanually  recorded

Special Sampling Requirements Collection,_ Storagey T^andling) ;  If
stored nore than 0-10 hours, add mercuric "chloride or preservative anc!
refriqerate at 4^C.

References;

1.  Methods for Chemical Analysis of TJater and Hastes.   1971.
    national Environmental ?.e??earch Center, Cincinnati,  Oliio.

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                                 79
                              PHOSPHORUS


Name of neasurement_fTethocl:   Automated flingle  Reagent Method

Principal Detection Technirruet  Colorinetry

Purpoi^e_of Measurement  (Important^pplications) ;   Ambient surface
waters^ "dome stic and industrial\vastes, saline  waters.   (May also
prove applicable to sediments, sludges.)

Summary of .Method;  Ammonium molybd.ate and  potassium antinonyl
tartrate react in an acid medium with dilute  solutions  of phosphorus
to form an antinony-phosphomolybdate cormlex.   This complex is reduced
to an intensely blue-colored comple:: by ascorbic  acid.   The color in
proportional ot the phorsnhoruf? concentration.

A Technicon AutoAnalyzer is used with a colorimeter ecmipped vr.ith G50
nm filter.

Limitations;

    Range ofApplicability;  0.01  to 1.0  mg/1 phosphorus

    Interferences;  Iron in hi
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                                 80
Data Outputs;  Analog electrical siqnal recorded on strinchart.

Special_f!anpling_Reguirement Collection,_rtorage, Handling) ; Avoid
benthic^leposits during collection.If not analyzed sane day as
collected, preserve with mercuric chloride and refrigerate  at

References:

1.  Methods for Chemical Analysis of Water and Uastos.  1971.  EPA
    National Environmental Research Center, Cincinnati, Ohio.

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                                 81



                              PHOSPHORUS


Name of Measurement Method;     Automated Stannous Chloride Method

Principal Detection Technique;  Colorinetry

Purpose of f'-easurenont  (Important  Applications);  Ambient  surface
waters, don^stic and industrial wastes.   (May prove applicable  to
sedinent, sludge and alqae blooms).

nummary of Method;  Phosphorus is  determined by manually digesting  the
samples with ammonium persulfate and sulfuric acid to convert the
various forns of phosphorus to the orthophosnhate form; and
measurement of this orthophosphate on a Technicon AutoAnalyzer, using
(NIL Jjjfto^with SnCl reduction to form color complex.

Limitations;

    Range of App1i cab i1i ty;  0.01  to 1.0 mq/1 phosphorus

    Pitfalls; Special Precautions;  Avoid benthic deposits when
    acquiring sample.  Do not use  commercial detergents for cleansing
    apparatus.

Statistical Characteristics;  On a single laboratory, using surface
v;ater sanples at concentrations of 0.06, 0.11, 0.48, and 0.62 ing
P/liter, the standard deviation was +0.004  (AOC Laboratory).

    Tine of Measurement;  About 15 samples per hour

Calibration Requirements;  Requires standard solutions.  See Ref.   (1)
for details.

Data Outputs;  Analog electrical signal recorded on stripchart.

Special Sampling Requirements Collection, Storage, Handling); Avoid
benthic deposits wJion sampling.  If sample is not analyzed on day of
acquisition, preserve with mercuric chloride and refrigerate at

References;

1.  Methods for Chemical Analysis  of T7ator and Wastes. 1971.  EPA
    National Environmental Research Center, Cincinnati, Ohio.

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                                   82
                                 SILIPA

Principal Detraction Tehrr.irmo!
     ro Q-P 'Vi^rm. recent  (Important Applications) ;  Drinkinrr v.v.ter,
anhiont surface voters,  domestic ant1 industrial wastes, saline waters.
        of Kethod ;  A vell-n.ixed  sample in filtered throuah a 0.45u
nonbranc filter.  Tho filtrate, unon the addition of nolyhflate ion in
acidic fiolutior,  forns  a  qroftninh-i-ynllow color connlc:: proportional to
tho flirnolvGd silica in thn  sf^nlo.   Thf» color co"nlr>:: ir then
measured spectronhoton.etricnllv.

Linitations ;

    Range of Applicability;   2 to 25 mg/1 silica

    Interferences ;  Excessive color  or turbidity

            Characteristics;  Photonetric evaluations by the anino-
naphtho-sulfonic aci.d procedure  have an estimated precision of +0.10
nq/1 in tho range from 0  to  2  rvf/1  (APT.M) .   Photonetric evaluation of
the silica-riolyhdate color in  the range from 2 to 50 na/1 have an
estimated precision of approximately 4 percent of the rruantity of
silica neasured
Data Outputs ;  Analoa electrical  sirmal

References;

1.  Hethods for Chenical Analysis of T'Tater and Pastes .
    1P71.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  APT1" Standards.  1970.   Part  23, Atmospheric Analysis
    Method DC 5 9-6 8.

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                                    83


                            OIL AND GREASE

Name of Measurement Method;  Extraction/Gravimetric

Purpose of Measurement (Important Applications); Measures hexane-
extractable matter  (animal fats, non-volatile hydrocarbons, waxes,
grease$) in surface waters, industrial wastes, and domestic sewage.

Summary of Method;  The sample is acidified to a low pH  (4.3) and
extracted with hexane using a Soxhlet extraction.  The solvent is
evaporated from the separated extract and the residue weighed.

Limitations;
    Range of Applicability;  5 to 1000 mg/1 of extractahle matter

Statistical Characteristics;  Precision and accuracy data not
available from reference below.

Time of Measurement;  Not stated, but several hours.

Data Outputs;  Read fron analytical balance.

References;

1.  Methods for Chenical Analysis of Water and Wastes.
    1971.  EPA national Environmental Research Center,
    Cincinnati, Ohio.

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                                  84
                                  PH

Name of Measurement Method;  Electronic trie

Principal Detection Technique^  Electronetric

Purpose of Measurement  (Important Applications) ! DrinMnq water,
ambient surface water's, domestic and industrial waters, so linn  waters,

Summary of Method;  The pll of a sarmle is an eloctronotric
measurement, using either a glass electrode in combination x\'ith a
reference potential (saturated calomel electrode) or a combination
electrode (glass and reference).

Limitations:
    Range of Applicability;  Not stated, but assumed broad.

    Interferencest  Oil and grease.

Statistical Characteristics;  Forty-four analysts in twenty
laboratories analyzed six synthetic water samplers containing exact
concentrations of hydrogen-hydroxyl ions, with the following results;

Known Cone.           Precision as          Accuracy as
as pll units        Standard Deviation     Bias,      Bias,
                        pH units            %          pF units

    3.5                 0.10              -0.29      -0.01
    3.5                 0.11              -0.0
    7.1                 0.20              +1.01      +0.07
    7.2                 0.18              -0.03      -0.002
    8.0                 0.13              -0.12      -0.01
    8.0                 0.12              +0.16      +0.01

    Time of Measurement;  Not stated, but rapid  (several per hour)

Calibration Requirements;  Instrument must be initially standardized.

Comments by Users;  Pield pR measurements made with comparable
instruments are reliable.

Data Outputs;  Analog sianal displayed on meter.

Special Sampling Requirements (Collection, Storage, Handling)^
Analyze"~as soon as possible after collection.  No holdinq.

References;
1.  Hethods. for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Wastewater.  1971.  13th Edition.  American Public
    Health Association, Washinqton, D. C.

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                                   85
                              PHENOLICS

Name oj: f'eajsuremervt Method;  4-AAP Method with Distillation

Principal Detection Technique* t  Colorimetry

Purpose of Measurement  (Important Applications); Drinking water,
ambient surface waters, domestic ana industrial wastes, saline waters.

Summary of Method:  Phenolic materials react with 4-aminoantipyrine in
the presence of potassium ferricyanide at a pH of 10 to form a stable
reddish-brown colored antipyrine'dye.  The amount of color produced is
a function of the concentration of phenolic material.

Limitations:
    Range of App1icabi1ity;  5 to 1000 g/1 phenol  (with solvent
                             extraction)50 to 5000 g/1 phenol  (without
                             solvent extraction)

    Interferences;  None stated.  pH must be controlled

    Pitfalls; Special Precautions;  Method does not differentiate
    different types of phenolic materials.

Statistical Characteristics;  The following results were obtained by
analysts at six laboratories:

             Known                Standard Deviation
         Concentration                (ug/1) with
           of Phnnol                   Solvent
              (g/1)                    Extraction	

              9.6                        0.99
             48.3                        3.1
             93.5                        4.2

Calibration Requirements;  Color response varies with type of  phenolic
material.Phenol is used as standard.

Data Qutnuts;  Meter reading (analog electrical signal).

References;

1.  Methods for Chenical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Uastowater.  1971.  13th Edition.  American Public

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                               8G
Health Association,  TTaRhin
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                                  87
                                SOLIDS
                              (DISSOLVED)

Principal Detection Technique:  Filtration/Hravinetric

Purpose of Iteasurenent  (Important Applications)^ Ambient surface
waters, domestic and industrial wastes,saline waters.

Summary of Method;  A well mixed sample is filtered through a standard
glass fiber filter.  The filtrate is evaporated and dried to constant
weiqht at 180°C.

Limitations;

    Range of Applicability;   10 to 20,000 mg/1 solids

    Pitfalls; Special Precautions:  Mineralized waters containing
    calcium, magnesium, chloride and/or sulfate may be hygroscopic and
    will require prolonged drying and quick weighinrr.  Samples
    containing bicarbonates require careful dryina at 180 C to insure
    conversion to the carbonate.

Statistical Characteristics:  Precision data not available at this
time.  References belovr.  Accuracy data on actual sample cannot be
obtained.

Data Outputs:  Read from analytical balance.

SpecialSampling Requirements  (Collection. Storage, Handling);
Sample should be analyzed as  soon as possible after acquisition.

References:
1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

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                                  88


                                SOLIDS
                              (SUSPENDED)

Principal Detection Technique;  Filtration/nravimotric

Purpose of Measurement  (Important Applications) ; Ambient surface
waters, donestic and industrial wastes, saline waters.

Summary of Method;  A well-mixed sample is filtered through a  standard
glass fiber filter, and the residue retained on the filter is  dried  to
constant weight at 103-105°C.  Non-filterable solids are defined as
those solids which are retained by a standard glass fiber filter and
dried to constant weiqht at 103-105°C.

Limitations:
    Range of Applicability;  20 to '20,000 mg/1 solids

    Pitfalls; Special Precautions;  Too much residue on the filter
         entrap water and require polonged drying .
Statistical Characteristics t  Reproduction of the data not available
at this time.  Accuracy data on actual samples cannot be obtained.

Data Outputs;  Analytical balance readimr.

Special Sampling Requirements (Collection, Storage, Handling) ;
Preservation of sample is not practical; analysis should begin as soon
as practical.  Non-homoqeneous particulates  (sticks, fish, etc.)
should be excluded from sample.

References;
1.  .Methods for Chemical Analysis of Water and T-Jastes.
    1971.  EPA national Environmental Research Center,
    Cincinnati, Ohio.

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                                 89
                                SOLIDS
                                (TOTAL)

Principal Detection Technique:  Filtration/Gravimetric

Purpose of Measurement  (Important Applications); Ambient surface
waters, domestic and industrial wastes, saline waters.

Summary of Method;  A well mixed aliquot of the test sample is
quantitatively transferred to a pre-v»eighed evaporating dish and
evaporated to dryness at 103-105°C.   (Total Solids are defined as the
sum of the homogeneous  suspended and dissolved materials in a sample),

Limitations;
    Range of Applicability;  10 to 30,000 mg/1 solids

    Pitfalls; Special Precautions;  Floating oils and greases, if
    present, should be dispersed with a blender.  Large floating
    particles should be excluded.

Statistical Characteristics;  Precision and accuracy data not
available at this time.

Data Outputs;  Read from analytical balance

References;

1.  Methods for Chenical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

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                                  90
                                SOLIDP
                               (VOLATILE)

Principal Detection Technique;  Ignition/Filtration

Purpose of Measurement  (Jimnortrint Applications) ; The  test  is  useful  in
obtaining a rbucrh annroximation of the amount of oraanic matter
present in the solid fraction of sewage, activated sludge,  industrial
v/astes, or bottom sediments.

Summary of "ethod;  The residue obtained fron the deternination  o^
total, suspended, or dissolved solids is ignited at 550°C  in  a muffle
furnace.  The loss of weiaht on ignition is renortect  as mg/1  volatile
solids.

Linitations;

    Pitfalls; Special Precautions:  The test is subject to  nany  errors
    due to loss of water of crystallization, loss of  volatile ornanic
    natter prior to combustion, incomplete oxidation  of certain
    conple:: orrranics, and decomposition of mineral snlts during
    combustion.

Statistical Characteristics:
    Renroducibility^  A collaborati^'e studv involving  three
    laboratories examining four samples by means of  ten  replicates
    showed a standard deviation of +11 pa/1 at  170 mg/1  volatile
    solids concentration.

Data Outputs;  Read from analytical balance.

References:
1.  Methods for Chemical Analysis o^ Tator and !7antes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    "antevrater.  1971.  13th Edition, American
    Public Health Association, T'anhinaton, P. C,

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                                  91
Principal Detection Technique;  Colorinotry

Purpose of Measurement  (Important Applications)! DrinMng water,
ambient surface waters, domestic and industrial was t e s.

Summary of Method:  Sulfate ion is converted to a barium sulfate
suspension under controlled conditions.  The resulting turbidity  ic
determined by a photoelectric colorimeter or spectophotoneter and
compared to a curve prepared from standard sulfate solutions.

Limitations:
    Ranae of Applic_a_biIj^ty;  All concentrations of sulfate  ion.
    Higher concentrations" must be diluted such that sample  aliquot
    contain less than 40 nq/1 sulfate.

    Interferences:  Suspended matter and color interfere.   Correct by
    running blanks from which the barium chloride has been  omitted.

Statistical Characteristics t  Thirty-four analysts in 16 laboratories
analyzed six synthetic water samples containing exact concentrations
of inorganic sulfate with the following results:

Known Cone, of       Precision as                Accuracv as
    Sulfate       Standard Deviation          Bias          Bias
    ma/liter           mg/liter                 %        ma/liter

      8.6               2.30                  -3.72         -.3
      9.2               1.70                  -0.26         -.8
    110.n               7.86                  -3.01       -3.3
    122.0               7.50                  -3.37       -4.1
    188.0               9.58                  •'•P.O/'         +.1
    199.0              11.8                   -1.70        -3.4

Time of reasurenent;  Rapid

Data Outputs;  Meter output (analog electrical sianal), manually

References;

1.  Methods for Chemical Analysis of "ater an<3 Wastes.
    1971.  EPA national Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Uastewater.  1971.  13th Edition.  American
    Public Health Association, Washington, D. C.

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                                  92
                                RULFAT

Harm of veasurerient He they'll   Autonnted CMoranilate J'ethod

Principal Detection ^echnirme;   Color.lr.otrv

Purpose of '•eppurenent  (Important Applications):  AnMont surface
waters", donestic and industrial wastes,  saline waters.

Runr.nry of I'.et3iorT;  TTion  solid  bariun chloranilate in added to a
solution oontpinina sulfate,  bariun sulfate is procinitatec!, releasing
tho l\irrhly colored acid chloranilate ion.   The color intensity in the
resulting chloranilir acid is proportionate to the amount of sulfate
nresent.

I'ether* ernloy.n Technicon  AutoAnal^'zor vrith colorin.eter eruipned \rith
    nn filters.
Limitations;

    Range of Applicability;   10  to 400  mg/1 sulfate

    Interferences;  Calciiim,  aluninum,  iron interfere, but nay he
    removed by ion exchange.

Statistical Characteristics;   In a single laboratory (A<">c) , using
surface water samples at concentrations of 39,  111, 188, and 194 mg
SO^/1, the standard deviations were +0.6, +1.0,  +2.2, and +0.8,
respectively.

    Tine of Measurement;  About  15 samples per  hour.

Calibration Requirements;  Requires preparation of standard solutions
and calibration curve.

Data Output;  Analog electrical  signal, recorded on stripchart.

Special Sampling  Requirements  (Collection, Storage, Handling);
Refrigerate at*4  C.

References:
1.  Methods for Cherdcal Analysis  of  !
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                                  93
                                SULFIDE

Name of Measurement Method:  Titrinetrie  (Iodine) Method

Principal Detection Technique;  Titration

Purpose ofMeasurement^(ImportantApplications) ; Drinkinrr waters,
ambient surf ace vjaters7 Domestic ~and industrial wastes, saline waters,

Summary of Method;  Sulfides are stripped from the acidified  sample
withTan inert gas and collected in a zinc acetate solution.   Excess
iodine added to the zinc sulfide suspension reacts with the sulfide
under acidic conditions.  Thiosulfate is used to measure unreacted
iodine to indicate- the quantity of iodine consumed by  sulfide.

Limitations:
    Range of Applicability;  Concentrations of  sulfide above  one mg/1

    Interferences;  Sulfites, thiosulfates, hydrosulfates,  and other
    reel need sulfur compounds may interfere.

    P_it falls; Special Precautions;  Sample should have minimal contact
    with air or oxyqon.

Statistical Characteristics;  Precision and accuracy  for  this method
have not been determined, but it is claimed that the  iodimetric
titration of the zinc sulfide is quite accurate.

Djita Outputst  Not stated.  Assumed to be visual observations manually
recorded.

Special Sampling Requirements (Collection, Storacre, Handling);
Minimize contact with air.Preserve with zinc  acetate unless
analysis is performed, immediately.

References:
1.  Methods for Chemical Analysis of Water and Wastes.
    1971.  EPA National Environmental Research Center,
    Cincinnati, Ohio.

2.  Standard Methods for Examination of Water and
    Wastewater.  1971.  13th Edition.  American Public
    Health Association, Washington, D. C.

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                                  94
                              TURBIDITY

Name of Measurement net-hod;  Hephelometric

Principal Detection Techninue;  Photometric  (TTephelometer)

Purpose of Measurement  (Important Applications): This method is
applicable to surface and saline waters in the ""range of turbidity fron
0 to 40 Jackson units.

Summary of Method;  The nethod is based upon a comparison of the
intensity^ of liqht scattered hv the sample under definite conditions
with the intensity of light scattered by a standard reference
suspension.  The hiaher the intensity of scattered liaht, the higher
the turbidity.  Readinqs, in Jackson units, are made in a nepholometer
designed accordinq to specifications outlined in Reference cited
below.  A standard suspension of Formazin, also prepared under closely
defined conditions, is used as the turbidity reference suspension for
water because it is more reproducible than other types of standards
previously used for turbidity standards.  Desiqn criteria for
instruments are given in Reference.

Limitations: Range of Anplicability;  0 to 40 Jackson units.
    Sensitivity should be 0.02 units or less in waters havinq
    turbidity of less than one unit.

    Interferences;  Floating debris, coarse sediments, air bubbles, or
    colored material on solution may interfere.

    Pitfalls; Special Precautions;  Care should be exercised in
    selection of instrument that meets criteria given in Reference.

Statistical Characteristics:
    Accuracy and Precision:  Data not available at time of publication
    of Reference.

    Time of Measurement:  Not stated.  Assumed rapid but not
    instantaneous, because sufficient time must be allowed after
    shaking sample for air bubbles to disappear but not for all
    suspended particles to settle.  (Five minutes or less).

Calibration Requirements;  Follow instrument manufacturer's
recommendations,but reliance on manufacturer's solid scatterinq
standard is not always acceptable. ' See Reference below.

Data Outputs:  Photoelectric detector (analog sianal) with ammeter
readout.

Special Sampling Requirements (Collection, Storage, Handling);

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                                  95
Samples taken for turbidity measurements should be analyzed as soon as
possible.  Preservation of samples is not recommended.

References:
1.  .Methods for Chemical Analysis of Water and Wastes.  1971.  EPA
    National Environmental Research Center, Cincinnati, Ohio.

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                             TEMPERATURE
Principal Detection Technique;  Thermonetry

Purpose of Measurement  (Important Applications):  Drinkinq waters,
ambient surface v/aters, cloncstic and industrial wastes,  saline waters.

Summary of Methodt  Temperature measurements nay be made x-rith any good
grade of mercury-filleri or dial-type centigrade thermometer, or  a
thermistor.

Limitations;

    Range of Applicability;  Thermometers or thermistors can be
    obtained to give valid results over almost any ranne of ambient
    temperatures.


    Pitfalls; rnecial Precautions;  Measurement device should be
    checked against a precision thermometer certified by the National
    Bureau of Standards.

Statistical Characteristics;  There is no accentable procedure for
determining the precision and accuracy of this test.

Time of Measurement;  Rapid

Data Outputs;  Usually a visual reading manually recorded.
Instruments producing analog voltages automatically recorded are
readily available.

References;

1.  i"ethods for Chemical Analysis of Water and Vastcs.   1971.  EPA
    National Environmental Research Center, Cincinnati,  Ohio.

2.  Standard Methods for Examination of TTater anc? Nastewater.  1971.
    13th Edition.  American Public Health Association, Washington, D.C

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                                 97
                            THRESHOLD ODOR
Name of Measurement Method;  Consistent Scries Method

Principal Detection Technique*;  Personal Sensing

Purpose of Measurement  (Important Applications);  This method is
applicable to the determination of threshold odor or finished waters.
surface waters, domestic and industrial wastes, and saline waters.

Summary of Method!  The sample of water is diluted with odor-free
water until a dilution that is of the least definitely perceptible
odor to each tester is found.  The resulting ratio by which the sample
has been diluted is called the "threshold odor number"  (T.O.).

Peoole vary widely as to odor sensitivity, and even the same person
will not be consistent in the concentrations he can detect from day to
day.  Therefore, panels of not less then five persons, and preferably
10 or more, are recommended to overcome the variability of usinq one
observer.

As an absolute minimum, two persons are necessary:  one to make the
sample dilutions and one to determine the threshold odor.

Limitations t

    Range of Applicability;  Highly odorous samples are reduced in
    concentration proportionately before being tested.  Thus, the
    method is applicable to samples ranging from nearly odorless
    natural waters to industrial wastes with threshold odor numbers in
    the thousands.

    Interferences;  Chlorine in tap water and some wastewaters may
    affect results.  It is sometimes desirable to determine the odor
    of the chlorinated sample and the same sample after removal of the
    chlorine.  Removal can be accomplished by use of sodium
    thiosulfate in stoichiometric proportions.

    Pitfalls? Special Precautions;  It is important to check a blank
    to which a similar amount of a dechlorinating agent has been added
    to determine if any odor has been imparted.  Such odor usually
    disappears upon standing if excess reagent has not been added.


Statistical Characteristics;

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                                  98


    Accuracy and Precision;  Data not  available  at  the  tine of
    publication of Reference cited below.

    Tine of 1 Teasurenent_s:  Several hours.

Calibration Recniirenents;  See Reference

Data Outputs;  Personal detection of odor

Special Sampling Requirements  (Collection,  Storage,  Handling);
Water sampTos ~"must bo collected in glass bottles \*ith glass or Teflon
lined closures.  Plastic containers are not reliable for odor samples
and nust not be used.  Complete tent as soon as  possible after sannle
collection.  If storage is necessary,  fill  container to the top and
refrigerate.

References;

1.  .Methods for Chenical Analysis of TJater  and Wastes.
    1971.  EP7\. National Environmental  Research Center,
    Cincinnati, Ohio.

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                                 99
                         SPECIFIC CONDUCTANCE
Principal Detection Technique;  Electrometric

Purpose of Measurement  (Important Applications) ;   Drinking  water,
ambient surface waters, domestic and  industrial wastes,  saline  waters.

Summary of Method; The  specific conductance of a  sample  is  measured by
use of a self-contained conductivity  meter, Wheatstone bridged-type, or
equivalent.  Samples are preferably analyzed at 25°C.  If not,
temperature corrections are made and  results reported at 25°C.

Statistical Characteristics;  Forty-one  analysts  in  17 laboratories
analyzed six synthetic  water samples  containing known concentrations
of inorganic salts, with hnown values of specific conductance.   The
followino results were  obtained;
Known Cone, of
Specific Conductance
 mhos/en

         ion
         106
         80S
         848
        1640
        1710
Precision as
Standard Deviation
 mhos/cm

      7.55
      8.14
     66.1
     79.6
    106
    119
     7\ccuracv as
  Bias          Bias,
    %         mhos/cm
•2.02
•0.76
•3.63
•4.54
•r>.36
•5.08
 -2.0
 -0.8
-29.3
-3H.5
-37.9
In a single laboratory  (7*OC) , using surface water  sanples with  an
average conductivity of 536 mhos/en at 25°C,  the standard deviation
was +6.

Calibration Requirements;  Peter reading  (analog electrical  signal,
manually recorded).

References;

1.  Methods for Chemical Analysis of Water and \7astes.   1971.   FPA
    National Environmental Research Center, Cincinnati,  Ohio.

2.  Standard Methods for Examination of Water and  T-Tantevater.   1971.
    13th Edition.  American Public Health Association, Washington,  D.C.

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                                 100
           IDrNTIFICATTON OF WTATJTFRTD OR I^TT-TTATITFREn  OILS



Medium;  V7ater, sediments, and tissue

Name of fleasurenent Method;  Woods Hole Oceanographic  Institute  .Method

Principal Detection Technirmes;  Has chromatography  (flane  ionization)

Purpose of Measurement  (Important.Applications);   Passive tagging of
oils                      J"

Summary of Method:  Oils are dissolved in carbon  disulfide  and
injected into the gas chromatocrraphy rn.  The  column is  50  feet  x 0.02
inches (open tubular support coated—SCOT)  column packed with  nonpolar
liquid silicone OTr-ini/ rated at 25,000 effective plates.   Oil
chronatoqrans are compared visually, and certain  features are
abstracted, tabulated and connared to the chronatograns  generated by
candidate unvreathernd oil sanples.

LinLtations;

    Interferences!  nannies which have undergone  unusual bacterial
    alteration contain hirrh levels of indiaenous  hvdrocarbons;
    nixtnres of oils and samples having under  gone umisually prolonged
    vreatherina reauire special treatment.

    Pitfalls; Special Precautions;  Tabulated  indices  of oils  that
    exhibit considerable v/eathering should  not be compared  directly
    vrith the indices of umreathered oils.

Statistical Chracteristics;

    Accuracy;  16 out of 17 unveathered oils used for  this  test
    acconpanied 35 sinulated oil spill samples.   Correct "definite
    correlation" was achieved in 74 percent of the cases, and  only one
    "probable correlation" vras incorrect.

    Precision;  Tabulated indices of individual oils remain quite
    stable firon column to column.

Calibration Requirements;  I'.ethod requires  frequent evaluation of
system performance by Tnjoctinq a standard  oil that yields  a
repeatable knovjn chromatogram.

Data Outputs;  Analoa instrument readina, graphically  recorded.

Special Sampling Requirements  (Collection,  Storage, Handling);

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                                   101
Sample should be refrigerated if held for more than a tow days.  Glass
or metal containers are required for sampling.

References;

1.  Zaifirion, 0. M. Bluner and J. Myers.  1972.  Correlation of Oils
    and Oil Products by Gas Chromatography.  !>7oods Hole Oceanographic
    Institution Report 62-55.

2.  Kawahara, Fred K.b  1969.  Laboratory Guide for the Identification
    of Petroleum Products, Federal Water Pollut5.on Control
    Administration, Division of Water Quality Research, Cincinnati,
    Ohio.

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                                  102
                            PF.TP.OLW  OILF
llane c>* Measurement Method;  Quantitative  Analysis  of  Oil in VTater
Diversion

Prj . ncir>a 1 De t e c t ion Techniones;   Infrared  Spectrophotoretry

Purpose of ^easnxenont  (Important Applications);   Identification of
the particular oTl^and  its cruantitative  determinations in the water
column are essential to proper I1"  monitor and  assess potential
biological damage renultinrr fron  oil  spill incidents.
	of Ilethod;  Halt and acid  added  to  the  sample  which is then
extracted v/ith carbon tetrachloricle or Freon  113,  using a separatory
funnel.  The extract is measured by infrared  spectrophotometry.

Limitations;

    Sensitivity;  Minimum detection limit   0.05  mg/1

    Interferences;  (Freon or CCl^) - Any solvent  extractable
organics.    ~~

    Pitfalls; Special Precautions:  Carbon  tetrachloride has a TLV of
    10 ng/1.:

Statistical Characteristics;

    Time of .Measurement:

Ca 1 ibr at ion Renu i rements;  Freon is not  usable  for preparing II?
standards of heavy oils  (2).

Comments by Users;  Carbon tetrachloride is more efficient than Freon
ll3 for extracting high concentrations of viscous  oils (1000
centistokes at 100 F) from water dispersion.

Data Outputs;  Analog instrumental reading.

Special ^amplinrr  Perruirements  (Collection,  Storage. Handling);
Sample is collected in a glass-stoppered bottle ancl acldified at the
tirae of collection.

Reference s_;

1.  Kawahara, Fred K.  1969.   Laboratory Guide for the Identification
    of Petroleum  Products, Federal Water Pollution Control
    Administration, Division of Water Quality Research, Cincinnati,
    Ohio.

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                                   103
Principal Detection Techniques:  Electron Canture, Cas Linuicl
Chronatography

Purpose of Measurement  (Important Applications);  To determine
presence and identification of pesticides in waters.

Summary of Method;  The v;ater sample is subject to multiple extraction
by ethyl ether/hexane.  The combined extracts are dried vrith anhydrous
sodium sulfate, evaporated under a stream of nitrogen, and further
concentrated by heating.  Aliquots of about 5  ul are used for the
initial electron capture gas liquid chronatorrraphy.  Further
identification may be necessary through microcoulometry of thin layer
chromatography.

Limitations;

    Interferences;  If interferences are indicated in initial
    chronatograns, it may be necessary to conduct a Floricil clean up
    on the extract.

Calibration Requirements;  See Reference

Data Outputs;  Analog electrical signal displayed as chromatogron.

Special Sampling Requirements  (Collection. Storage* Handling);  Sampling
collection glassware should be scrupulously cleaned.  If storage is
necessary samples should be kept in cool, dark place or preferably in
a refrigerator.

Reference;

1.  Thompson, J. F. (Editor).  1971.  Analysis of Pesticide Residues
    in Human and Environmental Samples.  EPA Primate and Pesticides
    Effects Laboratory, Perrine, Florida.

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                                  104
                              PESTICIDES
          Parameter(s) Measured;  Organochlorine Pesticides
    Aldrin
    BCH
    Chlordane
    ODD
    DDE
DDT
Dieldrin
Endosulfan
Endrin
Heptachlor
Heptachlor Epoxide
Chlorodane (tech)
riethexychlor
Perthane
Sulphenone
   under some
  circumstances;

Keithane
Strobane
Toxaphene
Principal Detection Techniques:
              Electron Capture  (EC)
              Microcoulometric Titration (MC)
              Electrolytic Conductivity (ECD)
Purpose of :ieasurement_(Important Applications);  For identification
and quantitation of various organocnlorine pesticides, certain
degradations products, and related compounds.

Summary of Method;  The method offers several analytical alternatives,
depending on the complexity of pesticides in the sample and the nature
and amounts of interferences.  In general, the pesticides in the
aqueous sample are extracted by organic solvents.   (e.g., hexane or
hexane/ethyl ether mixtures), and the extract concentrated by careful
evaporation of the solvent.  Removal of interferences (when necessary)
and pre-separation of pesticide mixtures are accomplished by column
chromatography, thin-layer chromatography, or liquid-liquid
partitioning.  Identification of pesticides in the mixture is made by
selective gas chromatographic separations through the use of two or
more unlike columns.  Detection and measurement of pesticides in the
sample are accomplished by electron capture, rnicrocoulometric, or
electrolytic conductivity techniques.  Quantitative results are
obtained by measurement of areas under peaks in the resulting
chromatogram.

    The reference cited describes the use of an effective solvent for
extraction of pesticides from aqueous samples.  It provides
information on the selection of appropriate clean-up procedures and
detectors for various types of pesticide mixtures, and suggests
techniques for confirming qualitative identifications.

Limitations:

    Range of Applicability;  Specific concentration ranges for various
    pesticides in environmental samples are not cited, but the
    detection capabilities of four types of detectors are indicated
    below.

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                                    105


    Sensitivity and Detection Limits;


         Electron Capture Detector picoqram  (110  grar1)
              quantities of many orqanochlorine pesticides

         f'icrocoulonetric Titration  5-20 ng o^ orqanochlorine
              pesticides

         Electrolytic Conductivity Sensitivity Detector
              2 to 3 tines as qreat as microooulometric procedure

         Flane Photonetric Detector  Sub-nanogram quantities
              of sulfur and phosnhorxis

         Sample responses less then tvo tines the detector  noise level
    (II) should be reported as negative; responses greater than  2F
    should bo quantified if possible.

    Interferences;  Polychlorinatod biphenyls; phthalate esters,
    organonhosphorus pesticides.  The presence of the  latter two
    classes of connounds are implicated in snnples that resnond to
    certain detection technioues but not to others.

    Pitfalls; Special Precautions;  Contaminants in solvents,
    reagents, glassware, and other equipnent nay yield results  that
    cause misinterpretation of chromatograms.

Statistical Characteristics;

    Accuracy;  Hot stated as percent relative error, but given  as
    percent recovery in Ref.

    Precision;  Results are given for four specific pesticides.  Those
    for aldrin are as follovrs;

                        Mean recovery       Precision  (ng/liter)


    llo cleanup


Florisil cleanup


Calibration Requirements;  Procedtiros are detailed in Reference.
(ng/liter)
10.42
79.00
17.00
64.54
Overall Sinqle Cperatio
4.86
32.06
9.13
27.16
2.59
20.19
3.48
R.02

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                                      106
Connonts by Usf»rs;  Reconnendod  for  une  only b",  or under close
supervision of, experienced residue  analysts.

Data Outputs;  Analog electrical  signals recorded on strincharts.

Special Sampling Reguirenents  (Collection,  fltorago/ ITand.ling);  T^ide-nouth
ipllnq
:h Tef
bottlon vrith Teflon-lined  screv  caps  are used for sanplo collection.

References;

1.  !*ethoc!s for Organic Pesticides  in Uater and WaFtevater.  1P71.
    EP.A National Environmental Research Center, Cincinnati, Ohio.

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2.  BIOLOGICAL  METHODS

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                                    107
                     PHYTOPLANKTON AND PERIPHYTON
                    CELL COUNTS AND IDENTIFICATION
Principal Detection Technique;   Visual Observation

Purpose of Measurement  (Important Applications);  Provide information
on the standing crops; indicator organisms and community diversity.
»
Summary of Method;  A 24x60 nun, No. 1 coverglass is placed diagonally
across the S-R cell and a large bore pipet or eyedropper is used to
transfer a one/ml aliquot of a well-mixed sample into the open corner
of the Sedgwick-Rafter chamber.  The S-R cell is allowed to stand for
at least 15 minutes to permit settling.  The analysis may then proceed
in either of two ways:  (1) depending on the density of organisms, two
to four "strips" the length of the cells are examined and all forms
that are totally or partially covered by the whipple-grid are
enumerated; or  (2) a minimum of ten random whipple fields are examined
in at least two identically prepared S-R cells  and forms that are
totally or partially covered by the whipple grid are enumerated.

Limitations;

    Range of Applicability;  Phytoplankton and  periphyton

    Interferences:  In samples where algae concentrations are extreme
    or where turbidity is high the sample must  be diluted or
    concentrated.

    Pitfalls; Special Precautions;  The depth of the counting chamber
    precludes the use of the 45x or lOOx objectives.  Collection of
    phtoplankton by nets or pumps is not recommended.

Statistical Characteristics;

    Precision;  Provided reasonably reproducible information when used
    with a calibrated microscope with an eyepiece measuring device.

    Time of Measurement;  1 hour

Calibration Requirements;  Each combination of  oculars and objectives
is calibrated against a stage micrometer.

Data Outputs;  Manual count

Special Sampling Requirements  (Collection, Storage, Handling);
The pretreatment of the sample depends on the concentration of
organisms present.  When phytoplankton densities are less then 500/ml,
approximately 6 liters of sample are required.

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                                    108
References;

1.   Biological Field and Laboratory Methods.  1972.  iiPA National
    Environmental Research Center, Analytical Quality Control
    Laboratory, Cincinnati, Ohio.  Preliminary Draft.

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                                    109
                         VOLUME OF PERIPHYTON
Principal Detection Technique;  Volumetric

Purpose of Measurement  (Important Applications);  Estimate of the
standing stock of periphyton.

Summary of .-lethod;  A known volume of water is added to a thoroughly
drained sample.  The difference between the volume of the periphyton
samples plus the added water and the volume of water alone is the
volume of the total amount of periphyton in the sample.

Limitations;
    Range of Applicability;  Periphyton/ only when large growths of
    periphyton permit removal of excess water readily.

    Pitfalls; Special Precautions;  Excess fluid should be removed
    from the sample.

Calibration Requirements;  Graduated cylinder should be used for
volume measurements.~

Data Outputs;  Volume measurement (visual)

Special Sampling Requirements (Collection/ Storage,  Handling);   The
preferred preservative is neutral or slightly basic formalin.

References;

1.  Biological Field and Laboratory Method.  1972.  EPA National
    Environmental Research Center, Analytical Quality Control
    Laboratory, Cincinnati, Ohio.  Preliminary Draft.

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                                   110
           CELL COUNTS AND IDENTIFICATION OF  PHYTOPLANKTON
Principal Detection Techniques;  Visual observation

Purpose of Measurement  (Important Applications);   Provides  information
on standing crop^indicator organisms and community diversity.

Summary of Method;  The sample is introduced  into  cells which  are
precisely matched glass slides with a finely-ruled grid on  a counting
plate fitted with a specially ground cover slip.  All forms which fall
within the griclded area of the cell are identified and counted.  The
number of the various organisms found in the  gridded  area of the cell
is multiplied by the appropriate factor to obtain  the count.

Limitations;

    Pitfalls; Special Precautions;  The analyst  is advised  to  follow
    carefully the specific directions accompanying the chamber or
    cell.  Collection of phytoplankton by nets or  pumps is  not
    recommended.  For statistical purposes a  normal sample  must be
    either concentrated or a large number of  mounts per sample should
    be examined.

Statistical Characteristics;

    Time or Measurement;  1 hour

Data C)utputs;  Manual count

Special Sampling Requirements  (Collection/ Storage, Handling;
Depends upon the location and type of sample. See  Reference.  Samples
must be concentrated when densities are below 500/ml. Approximately 6
liters of sample are required.

References;

1.  Biological Field and Laboratory Methods.  1972.   EPA National
    Environmental Research Center, Analytical Quality Control
    Laboratory, Cincinnati, Ohio.   (Preliminary  Draft).

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                                     Ill
           CELL COUNTS AND IDENTIFICATION OF PHYTOPLANKTON
Principal Detection Technique;   Visual observation (Filter Method)

Purpose of Measurement (Important Applications);  Provides information
on standing crop^indicator organisms and community diversity.  Method
permits the use of high magnification for enumeration of small
plankton.

Summary of Method;  A water sample of known volume is passed through
the membrane filter under a vacuum.  The filter is allowed to clear
and the organisms enumerated.  The occurrence of each species in 30
random fields is recorded and multiplied by a conversion factor to
obtain the total count of each species.

    Interferences;  Significant amounts of suspended matter may
    obscure or crush the organisms.

    Pitfalls; Special Precautions;  In coastal and marine waters the
    filter is rinsed with distilled water to remove salt.  Collection
    of phytoplankton by nets or pumps is not recommended.

Statistical Characteristics;
    Precision;  + 16% CL

    Time of Measurement;  Relatively rapid processing of samples, (1
    hour) although total time required for the complete analysis is
    48-50 hours.

Calibration Requirements;  Use stage and ocular micrometers

Data Outputs;  Manual count

Special Sampling Requirements (Collection, Storage, Handling;
Depends upon the location and type of sample taken.See Reference.
Samples must be concentrated when phytoplankton densities are less
than 500/ml.  Approximately 6 liters of sample are required.

References;

1.  Biological Field and Laboratory Methods.  1972.  EPA National
    Environmental Research Center, Analytical Duality Control
    Laboratory, Cincinnati, Ohio.   (Preliminary Draft).

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                                  112
           CULL COUNTS AND IDENTIFICATION OF PilYTOPWviJKTOIJ
Principal Detection Technique;  Visual observation   (Countiny Chamber Method^

Purpose of Measurement  (Important Applications);  Provides information
on algal standing crop^indicator organisms and  community diversity.

Summary of Method;  The method uses an inverted microscope in
conjunction with a cylindrical counting chamber with a clear glass
bottom.  A sample is transferred to the counting chamber and allowed
to settle.  The chamber is placed on the microscope stage and examined
using either the 20x, 45x or  lOOx oil immersion lens.  Either the
strip or random field counts  are made.

Limitations;

    Interferences;  None identified

    Pitfalls; Special Precautions;  Collection  of phytoplankton by
    nets or pumps is not recommended.

Statistical Characteristics:

    Time of Measurement;  4 hours per 10 mm of  sample height in the
    chamber is allowed for sedimentation.  Count requires 1 hour.

Calibration Requirements;  Use stage and ocular micrometers

Data Outputs;  Manual count

Special Sampling Requirements  (Collection, Storage, Handling);
Depends upon the location and type or sample taken.  Se«* Rpfpr*»nrw».

References;

1.  Biological Field and Laboratory Methods.   1972.  UFA National
    Environmental Research Center, Analytical  Duality Control
    Laboratory, Cincinnati, Ohio.   (Preliminary Draft).

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                                  113
           CELL VOLUME ESTIMATES OF PLANKTON AND PERIPHYTOH
Principal Detection Technique;  Visual

Purpose of Measurement  (Important Applications);  Standing crop
estimate gives an indication as to water productivity.

Summary of Method;  An aliquot of sample is concentrated and examined
wet at a 1000 x magnification with a microscope equipped with a
calibrated ocular micrometer.  The optical measurements are made with
the micrometer.  The average volume per organism is determined and
multiplied by the number of organisms per millilitar.

Limitations;

    Range of Applicability;  Phytoplankton, bacteria, periphyton

Calibration Requirements;  The exact magnification with any set of
oculars must be calibrated.

Data Outputs;  Visual examination

Special Sampling Requirement  (Collection, Storage, Handling):
The preferred preservative is neutral or slightly basic formalin.

Reference:
1.  Biological Field and Laboratory Methods,  1972.  LJPA National
    Environmental Research Center, Analytical Duality Control
    Laboratory, Cincinnati, Ohio.  {Preliminary Draft).

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                                   114
                     PERIPIIYTON ATID Pr^
                         (Species Composition)
Principal Detection Technique;  Visual examination

Purpose of Mea5urement  (Important Application);  Thr  identification  of
individual species of phytoplankton nay provide  information on
indicator organisms, species diversity, and therefore the  decree  of
pollution.

£unnary of "nthocl;  Folio'v.rv an initial examination  of  the sample to
obtain" rr> -rtirato of population etc., the phytoplankters  are identified
to the desired taxonomic level and tallied under a  standard system.


Limitations;

    Range of Applicability;  Plankton, periphyton

    Pitfalls; Pnocial Precautions!  Examination  if?  preferably done
    before sample is preserved.The beginner  is strongly  vjarned
    against the deceiving and nonvalid simplicity often  found in  the
    identification of plankton.

Statistical Characteristics;

    Time of .Measurement;  1 hour

Data Outputs;  Numbers of species and the number of individuals per
species

Special .Sampling Requirement  (Collection, Storage,  Hand ling);
In cases; \7liere it has been shown that preservation  has no  effects on
the identification of organisms, a preservative  should be  used if
storage is necessary.

References:

1.  Biological Field and Laboratory I'ethods.   1972.   EPA national
    Environmental Research Center /Analytical  Quality Control
    Laboratory, Cincinnati, Ohio.   (Preliminary  Draft).

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                                 115


                    DIATOM SPECIES IDENTIFICATION



Principal Detection Technique;  Visual Observation

Purpose of Measurement  (Important Applications);  Provides information
on indicator organisms .and community diversity.

Summary of Method;  The diatoms are concentrated by centrifugation
followed by sedimentation.  The diatom concentrate is placed on an 18
mnrcpverglass and dried on a hotplate at 95°C followed by the
oxidization of organic matter.  The hot coverglass is inverted and
placed on a drop of hyrax on a 25x75 mm microscope slide.  A
protective coating of clear lacquer is sprayed on the frosted end of
the slide and the excess hyrax is scraped from around the coverglass.
Two hundred fifty diatoms are identified and counted at high
magnification under oil.  If the slide has very feu diatoms the
analysis is limited to the number of cells encounted in 45 minutes of
scanning.

Limitations;

    Range of Applicability;  Diatoms

    Interferences;  Silt

    Pitfalls; Special Precautions;  If the dried sample is obscured by
    soluble solids,the sample should be washed with distilled water.

Statistical Characteristics;

    Time of Measurements;  1 hour

Calibration Requirements;  Ocular and stage micrometer

Data Chitputs;  Number of individuals per species

Special Sampling Requirements (Collections, Storage, Handling);
Depends upon the location and type of sample taken/ see Reference.  If
plankton counts are less than 1000 per ml, the diatoms should be
concentrated from a larger volume of sample (one to five liters) by
allowing them to settle out.

References;

1.  Biological Field and Laboratory Methods.  1972.  LPA National
    Environmental Research Center, Analytical Duality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                   116
           CELT. COUIITF  A*TD  IPniTIFIC^TIOlT OF PITyTOPLATT7rTO!I
Principal Detection Techniques;   Visual observation

Purpose of Measurement  (Important Applications);   Provides information
on standing crop indicator" organisms and community diversity.

Summary of Method;  An  aliquot  of well  nixed sample is introduced into
one of the '2x"5~~nm channels  on either side of the  circular Palner-
Ilaloney Hannop lank ton cell  <-:'.th  cover slip in place.   After 10
minutes, the sanple is  examined  under the high-dry objective  (45x)
and at least 20 v;hippie fields  arc counted.

Limitations;

    Range of Applicability!  The circular chamber used v/as especially
    designed for enumerating nannoplanJ'ton with a hiqh-dry obiective
    (45:0.

    Pitfalls; Special Precaiitions;   The cell should not be used for
    routine counting unless the  samnles have counts exceeding
    20,000/ml.  Collection  of phytoplankton by net or punps is not
    recommended.

Statistical Characteristics;

    Time of Iteasurenent;  1 hour

Calibratic-n Requirements;   The microscope is calibrated using an
ocular and stage micrometer,


Comments by Users;

Data Outputs;  .Manual counts

Special Sampling Requirement  (Collection, Storage, Handling);
Depends upon the location and type  of" sample.  See Reference.  .Camples
must be concentrated when densities are below 500/ml.   Approximately G
liters of sample are reauired.

References;

1.  Biological Field and Laboratory Methods.   1972.   EPA National
    Environmental Research  Center,  Analytical  Quality Control
    Laboratory, Cincinnati, Ohio.   (Preliminary Draft).

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                                  117
                    CHLOROPHYLL a OF
                              (In vitro)
Name of feanuromnnt Method;   Monochromatic Method

Principal Detection Technique*;  Fnectrophotometric

Purpose of Measurement  (Important Applications);  All algae contain
chlorophyll a.  Measurement of this pigmpir.t can yield an estimate o*
standing crop.

Summary of .Method t  Chlorophyll a can he cr.tinatnd independently of
the other chlorophylls by measuring the optical density of the pigment
extract of C>C<5 nr only, and inserting it into the followino equation:
C*,= 13.4D665.   (P665 is the optical density, corrected for turbidity
bv substractinn the OP nm blank).
                     7eo
Linitations;

    Pitfalls; Special Precautionn;  Precautions should bo taJ^en to
    min in i zo evapora tio n.  Phebphyt in    a natural dooradation product
    of chlorophyll, has an absorption peaJ- in the sane region of the
    visible spectrum as chlorophyll a and could be a source of error
    in chlorophyll determinations.

Calibration Requirements;  No unusual requirements

Data Outputs;  Analog or digital electrical signal displayed on peter,
on tape or recorder.

Special Sampling Requirements  (Collection, Ptoragn. Handlina);
Spec
The
    type of sampling equipment used is highly dependent upon where and
how the sample is being taJ'en.  If the analysis will be delaved, storo
the sample frozen in the darh to avoid, photochemical breakdown of tho
chlorophyll.

References;

1.  Biological Field and Laboratory .Methods.  1972,  EPA rational
    Environmental Research Center, Analytical Puality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                  118
                    CHLOROPHYLL a OF PHYTOPLANKTON
Principal Detection Technique;  Spectrophotometric

Purpose of Measurement  (Important Applications);  All algae contain
chlorophyll a_.  Measurement of this pigment can yield an estimate of
standing crop.

Summary of Method;  Chlorophyll a_ can be estimated independently of
the other chlorophylls by measuring the optical density of the pigment
extract of 665 nm only, and inserting it into the following equation:
Ca,= 13.41)4,6,*-  (D^^r is the optical density, corrected for turbidity by
subtracting the OD 750 nm blank).

Limitations;

    Pitfalls; Special Precautions;  Precautions should be taken to
    minimize evaporation.Pheophytin    a natural degradation product
    of chlorophyll, has an absorption peak in the same region of the
    visible spectrum as chlorophyll a, and could be a source of error
    in chlorophyll determinations.

Calibration Requirements;   No unusual requirements.

Data Outputs;  Analog or digital electrical signal displayed on meter,
on tape or recorder.

Special Sampling Requirements (Collection, Storage, Handling);
Tho type of sampling equipment used is highly dependent upon where and
how the sample is being taken.  If the analysis will be delayed, store
the sample frozen in the dark to avoid photochemical breakdown of the
chlorophyll.

References;

1.  Biological Field and Laboratory Methods.  1972.  EPA National
    Environmental Research Center, Analytical Uuality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                119
            ZOOPLANKTOH VOLUME AND SPECIES IDENTIFICATION
Purpose of iMeasurement  (Important Applications)  ileasurement of
zooplankton.  Volume provides an index to the standing crop  (biomass)
of natural zooplankton population.

Summary of Method;  The sample is screened, placed into a conical
container graduated in milliliters, allowed to settle for five
minutes, and the settled volume is recorded.  The sample is then
stirred and a one ml subsample is withdrawn from the container with a
Stempel pipette.  The subsample is examined under a dissecting
microscope for enumeration of zooplankton.

Limitations;

    Range of Applicabilityt  Rotifera, cladocera, copepods and other
         large zooplankton forms.

Statistical Characteristics;

    Tine of Measurement;  Less than 1 hour

Data Outputs;  Visual observation, manual reading

Special Sampling Requirements (Collection, Storage, Handling);
Depends upon location and type of sample taken.  See Reference.

Reference;

1.  Biological Field and Laboratory Methods.  1972.  EPA National
    Environmental Research Center, Analytical Duality Control
    Laboratory, Cincinnati, Ohio.   (Preliminary Draft).

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                                120


                III SITU PRODUCTIVITY OF PHYTOPLAMKTOH



Principal Detection Technique;  Radioactive assay

Purpose of Measurement (Important Applications)!  Phytoplankton
productivity measurements indicate the rate of uptake of inorganic
carbon by phytoplankton during photosynthesis and are useful in
determining the effects of pollutants and nutrients on the aquatic
community.

Summary of Method;  A solution of radioactive carbonate is added to
light and dark bottles which have been filled with aarnples taken from
preselected depths in the euphotic zone.  Following in situ incubation
(up to four hours) the plankton is collected on a membrane filter,
dried in a desiccator, and assayed for radioactivity.  The quantity of
carbon fixed is proportional to the fraction of radioactive carbon
assimilated.  If measurements are required for the entire photoperiod,
samples may be taken at overlapping four-hour periods or the data may
be adjusted based on the solar radiation profile.

Limitations;

    Range of Applicability;  Phytoplankton

    Pitfalls; Special Precautions;  Carbon in the filtered sample
    should yield the number of counts required for statistical
    significance.

Statistical Characteristics;

    Sensitivity;  Method is more sensitive than the oxygen method, but
    fails to account for the organic materials that leach from cells
    during the incubation period.

    Time of Measurement;  Sample should be incubated for at least two
    hours.

Comments by Users;  Method affords a direct measurement of carbon
uptake and measures only photosynthesis.

Data Outputs;  Instrument count

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                               121
References;

1.  Biological Field and Laboratory Methods.  1972.  EPA National
    Environmental Research Center/ Analytical quality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

2.  Standard Methods for the Examination of Water and Wastewater.
    1971.  13th Edition.  American Public Health Association,
    Washington, D. C.

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                                      122
                IN SITU PRODUCTIVITY OP PHYTOPLANKTON
Name and Measurement Method;  Oxygen method

Purpose of Measurement  (Important Applications);  Phytoplankton
productivity measurements indicate the rate of uptake of inorganic
carbon by phytoplankton during photosynthesis and are useful in
determining the effects of pollutants and nutrients on the aquatic
community.

Summary of Method!  Samples are taken from preselected depths in the
euphotic zone and placed in duplicate  clear, darkened and initial
analysis bottles.  The duplicate clear and darkened bottles are
suspended at the depth from which the samples are taken and allowed to
incubate.  At the end of the exposure period the dissolved oxygen of
the samples is measured.  The increase in oxygen concentration in the
light bottle during incubation represents net production and the loss
of oxygen in the dark bottles is an estimate of respiration.

Limitations;

    Range of Applicability;  Phytoplankton

    Pitfalls, Special Precautions;  The incubation period should not
    be long enough to allow oxygen gas bubbles to form in the clear
    bottles or dissolved oxygen to be depleted in the dark bottles.
    The solar radiation profile in addition to the photosynthetic rate
    during the incubation period should be used to adjust data to
    represent productivity for the entire photoperiod.

Statistical Characteristics;

    Time of Measurement;  Samples should be incubated for at least two
    hours.

Comments by Users;  Chief advantages of the oxygen method are that it
provides estimates of gross and net productivity and respiration and
that the analysis can be performed both with inexpensive laboratory
equipment and common reagents.

Data Outputs;  Visual observation (titrimetric)  or analog or digital
electrical signal (electrometric DO method).

Special Sampling Requirements (Collection, Storage/  Handling);
If the dissolved oxygen is not measured immediately, it should be
fixed and protected from direct sunlight.

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                                    123


References;

1.  Biological Field and Laboratory Methods.  1972.  EPA National
    Environmental Research Center, Analytical Duality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                    124


           PHYTOPLAUKTOIJ AND PLRIPHYTON SPECIES COMPOSITION



Phytoplankton and Periphyton Species Composition

Principal Detection Technique;  Visual examination

Purpose of Measurement (Important Applications);  The identification
of individual species of phytoplankton may provide information on
indicator organisms, species diversity, and therefore the degree of
pollution.

Summary of Method:  Following an initial examination of the sample to
obtain an estimate of population density, etc., the phytoplankton are
identified to the desired taxonomic level and tallied under a standard
system.

Limitations;

    Range of Applicability;  Plankton, periphyton

    Pitfalls; Special Precautions;  Examination is preferably done
    before sample is preserved.  The beginner is strongly warned
    against the deceiving and nonvalid simplicity often found in the
    identification of plankton.

Statistical Characteristics;

    Time of Measurement;   1 hour

Data Outputs;  Number of species and the number of individuals per
species


Special Sampling Requirements (Collection, Storage,^ Handling]I ;
In cases where it has been shown that preservation has no effects on
the identification of organisms, a preservative should be used if
storage is necessary.

References;

1.  Biological Field and Laboratory Methods.  1972.  EPA National
    Environmental Research Center, Analytical Quality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                    125
               CHLOROPHYLL a, b, AND c OF PHYTOPLANKTON
                            AND PERIPHYTON
Name of Measurement Method;  In Vitro - Trichromatic Method

Principal Detection Technique;  Spectrophotometrie

Purpose of Measurement  (Important Applications);  All algae contain
chlorophyll.Measurement of these pigments can yield an estimate of
standing crop and taxonomic congestion.

Summary of Method;  The sample is concentrated/ macerated, steeped in
90% acetone, at 4*C for 24 hours, and clarified by centrifugation.
The optical density of the decanted extract is determined at 750, 663,
645, and 630 nm using 90% acetone blank.  The 750 nm reading is used
to correct for turbidity and the other readings are inserted into
SCOR/UNESCO equations for calculating chlorophyll a, b, and c_.

Limitations;

    Pitfalls; Special Precautions;  Precautions should be taken to
    minimize evaporation.Phebphytin, a natural degradation product
    of chlorophyll, has an absorption peak in the same region of the
    visible spectrum as chlorophyll a and could be a source of error
    in chlorophyll determination.

Statistical Characteristics;

Calibration Requirements;  No unusual requirements.

Data Outputs;

Data Outputs;  Analog or digital electrical signal

Special Sampling Requirements (Collection, Storage, Handling);
If the analysis will be delayed, store fro2en.The stored samples
must be kept in the dark to avoid photochemical breakdown of the
chlorophyll.  Maximum storage time is 30 days.

References;

1.  Biological Field and Laboratory Methods.  1972.  LPA National
    L'nvironmental Research Center, Analytical Duality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                   126


                  CELL SURFACE AREA OF PHYTOPLANKTON



Principal Detection Technique;  Visual observation

Purpose of Measurement (Important Applications);  Determination of
cell surface area of phytoplankton is another indicator of plankton
abundance.

Summary of Method;  Measure the dimension of several representative
individuals of each major species  microscopically.  From the linear
dimensions compute the average surface area per species.  Multiply the
area per species by the number of organisms per mi Hi liter.

Limitations;

    Range of Applicability;  Phytoplankton.


Calibration Requirements;  Ocular and stage micrometric


Data Outputs;  Visual observation

Special Sampling Requirements (Collection, Storage, Handling);
Depends upon the location and type of sample taken, see Reference.

References:

1.  Biological Field and Laboratory Methods.  1972.  EPA National
    Environmental Research Center, Analytical Quality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                   127
                        BIOMAES OF MICROPHYTES



Name of Measurement Method;  Dryweight biomass

Principal Detection Techniques;  Gravimetric

Purpose of Measurement (Important Applications)!  Estimates of growth
rates related to pollution such as nutrient stimulation, can be
accomplished by standing crop estimates at predetermined intervals.

Summary of Method;  A sample is taV-en from a small defined area with
conspicuous borders.  The wet weight of material is obtained after the
plants have drained for a period of time.  The sample is then dried
for 24 hours at 105°C and reweiohed.  The dry weight of vegetation per
unit area is then calculated.

Data Outputs;  Mechanical scale reading

Special Sampling Requirements (Collection. Storage, Handling);
Before beginning a miantitative investioation it is desirable to have
a statistical study design which will assist in determining the best
sampling procedure, sampling area size, and number of samples.

References:

1.  Biological Field and Laboratory Methods.  1972.  FPA national
    Environmental Research Center, Analytical Quality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                    128
                        ALOAT, GROWTH POTENTIAL
Name of Measurement;  Bottle test method

Principal Detection Techniques;  Volumetric, Gravimetric, etc.
(depends on method used to indicate algal rrrowth).

Purposeof Measurement (Important Applications);  Alqal assays are
conducted to determine the effects of various discharges on the growth
of algao or to determine whether or not varionr, compounds or water
samples are toxic or inhibitory to algae,

Summary of M.ethnd;  7^fter inoculation of test organisms the sample is
incubated at 24 4- 2°C under cool white fluorescent lighting.  During
incubation, algaT grov/th is determiner! by cell counting, "or algal
hionass or the chlorophyll analynis methods at predetermined
intervals.  The collected data are used to determine the maximum
standing crop and the maximum specific growth rate.

Limitations;

    Pitfalls, Special.. Pro cautions;  T'Then trace nutrients are being
    studied special galssware such as Vycor or polycarbonate
    containers should be used.

Statistical Characteristics;

    Precision;  Excellent aareer.ent in data using this method was
    obtained by eight participating laboratories.

    Time of .Measurement!  The analysis continued until there is less
    than 5 percent per day increase in biomass.

Calibration Requirements;  Fee individual method in Reference cited
below.

Data Outputs;  Visual counts

Special Sampling Requirements  (Collection, Storage, Handling);
Sample pretreatment is~'"required when determinations are being made
6f  (1) growth limiting soluble nutrients or  (2) amount of algal
biomass then can be grown from all nutrients in waters.

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                                    129
References;

1.  Biological Field and Laboratory Methods.   1972.  FPA national
    Environmental Research Center, Analytical Quality Control
    Laboratory, Cincinnati, Ohio.  (Preliminary Draft).

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                                    130
                    CHLOROPHYLL a OF PlIYTOPLAiJiC'TON
                             (Fluorescence)
IJane of Measurement Ilethod;  In vivo method

Purpose of Measurement  (ImportantApplications);  All algae contain
chlorophyll a_.  Measurement of this pigment can yield an estirate of
standing crop.

Limitations;

    Rangu of Applicability;  Scale deflection should be between 15 and
    90 units.

    Interferences;  Phaeohytin

    Pitfalls; Special Precautions;  This method ia less efficient than
    the extraction method, yielding about one-tenth as much
    fluorescence per unit weight as the same amount in solution.

Statistical Characteristics;

    Sensitivity;  Method is more sensitive than the spectrophotometer
    method.

    Precision;  The precision of the method using natural population
    shows for ten samples, a maximum variation of 15 percent.

    Time of Measurement;  Less than 2 hours.

Calibration Requirements;  The fluorometer should be calibrated with a
chlorophyll extract which has been analyzed with a spectrofluorometer.

Comments by Users;

Data Outputs;  Analog or digital electrical signal

Special Sampling Requirements (Collection, Storage. Handling);
Prolonged storage or extracts(greater than 5 days;under
refrigeration and darkness should be avoided.  Fluorescence of
extracts kept at room temperatures and in light are stable for at
least ten hours.

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                                    131


Hcferences;

1.  Yentsch, C. G. and D. W. Menzel.  1963.  A Method for the
    Determination of Phytoplankton Chlorophyll and Phaeophytin by
    Fluorescense.  Deep-Sea Research 10;  221-281.

2.  Biological Field and Laboratory Methods.  1972.  iiPA National
    Environmental Research Center, Analytical Duality Control
    Laboratory, Cincinnati, Ohio.   (Preliminary Draft).

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3.  BIOASSAY PROCEDURES

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                                     132
                            3.  BIOASSAYS
    The purpose of this section is to discuss some available bioassay



procedures for determining "safe" levels of pollutants.  Bioassays are



essential to evaluate a qiven pollutant in terms ot existing water



quality, including environmental variables as well as pollution



already present.  Pertinent to this stance is the fact, that the



majority of specific pollution problems are those involving discharge



of unknown and variable composition.  Almost without exception, more



than one toxicant or stress is present.  It is believed that "safe"



levels of toxic pollutants, predicated upon bioassays with application



factors, adequately protect, aquatic life, if the basic physical



parameters, e.g., dissolved oxygen, temperature-, and pH are within the



limits recommended.  Tf the latter parameters are outside recommended



limits, appropriate alterations in the criteria for toxicants must be



made.







Acute ly^Lethal^Versus^^Sate^Cgncentrat ions








    Harmful effects of pollutants may be divided into a number of



categories which overlap somewhat; most could be described accurately



by using one or two of the following terms, all of which have good

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                                    133
dictionary meanings.  In the present context of toxicity to aquatic

life, the terms might be briefly defined as follows:



              t
    acute - involvinq a stimulus, severe enough to bring about a

response speedily, usually within four days for fish;




    subacute - involving a stimulus which is less severe than an acute

stimulus, and which produces a response in a longer time; may become

chronic




    chronic - involvinq a stimulus which is lingering or continues for

a long time; often signifying periods of about one-tenth of tne life

span or more;




    lethal - causing death, or sufficient to cause it, by direct

action;




    sublethal - below the level which directly causes death;




    cumulative - brought about, or increased in strength, by

successive additions.




    threshold - the point at which a physiological effect begins to be

produced.

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                                     134
    These and other terms have sometimes been used in careless or
conflicting ways, but they should be used with precision.  In general,
two broad categories of effects may be distinguished; acute toxicity,
which is usually lethal, and chronic toxicity, which may be lethal or
sublethal (1).

    Most of the available acute toxicity data are reported as the
median tolerance limit (TLm)  or median lethal concentration (LC50) .
Either symbol signifies the concentration at which 50 percent of the
test organisms survive within a specified time span, usually in 96
hours.  The customary 96-hour (four day)  time period is recommended as
adequate for most routine tests of acute toxicity with fish and the
lethal threshold concentration attained within this time is usually
reported (2).  Mortality tests of a week or longer are necessary at
times to determine the threshold concentrations.  The lethal threshold
concentration, as well as absence of any threshold concentration,
should be reported.  The lethal threshold concentration is more useful
for comparative purooses than the arbitrary LC50.

    This system of reporting is merely a convenient reference point
for expressing the acute lethal toxicity of a given toxicant tor the
test animal.   Usually, the "safe" levels (those which permit
reproduction, growth, and all other normal life-processes in the
fish*s natural habitat)  are much lower than the LCSO.

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                                     135
    Data on both lonq-term effects and safe levels are available for



only a few toxicants.  Some information is now becoming available on



the effect of toxicants on reproduction, a very important aspect of



all long-term toxicity tests.  Also important is the sensitivity of



the various life staqes of organisms.  Many organisms are most



sensitive in the larval, nymphal, molting, or fry stage; some may be



most sensitive in the egg and sperm stage.








    It would be desirable if a single, universal, rapid, biological



test could be used to directly measure sublethal effects of a



pollutant.  A large number of sublethal responses of fish have been



used, i.e.  specific physiological and biochemical changes in various



body systems, and histological studies.  A review of these shows that



unfortunately, no single test is meaningful for all kinds of



pollutants (3).








variabl,e_ConcentraMon








    Criteria for continuously acceptable concentrations must be lower



than the intermittent higher concentrations that may be reached



occasionally but briefly without causing damage.  One way in which to



resolve this difficulty is to use both maximum short-time



concentrations and also a more restrictive range of safe



concentrations for continuous exposure.  It is recognized that



extremes do limit organisms, but, within these extremes there are

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                                     137
ranges of concentration that can be tolerated, and are sate for
prolonged periods of exposure.  Most of r.h^> criteria recommended in
Volume I of this report are those thouqht to be safe for continuous
exposure.

    In field situations and industrial operations, average 21-hour
concentrations can be determined by using a small water pump to
collect small aliquots of a few ml every few minutes, or to collect a
slight trickle continuously.  After 21 hours, the sample is mixed and
analyzed and the concentration found represents the average
concentration.  Samples obtained this way are more reproducible and
easier to secure than t.he maximum instantaneous concentration.  But
average concentrations are of little significance if fish are killed
by a sharp peak of concentration; hence the need to determine maximum
concentrations.

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                                     138
                         METHODS OF BIOASSAY
    The use of bioassays to det^rmiac.- the  to::icitv of. «« material  or
waste  is the most effective and accurate  method of predicting or
assessing potential danger.  In use of a bioasray, no assumptions need
be made concern inq the chemical structure  or form of the pollutant,
nor does the investigator have to hnov; the constituent substances.
The effects of water quality on toxicii-y also rray be measured.  of a
toxicant in water, the more precise the assay can be.

li233say^_to_Determine_AcuteJ.Y_Lethal_Concent rations

    While there are many types of assays,  two are in general use: the
static bioassay in which the organisms are held in a -t-.ank of standing
test-solution, which may or may not be changed during the period of
the test; and the continuous flow or flow-through bioassay in which
the test solution is renewed continually,  occasionally the difference
between the two types is not very great, but sometimes one has clear
advantages over the other.

    A recommended outline of methods for routine bioassays is given by
"Standard Methods" (U) .  Methods for more  searching tests and for
research purposes have been critically reviewed  (3,5,6).  The items
which should be described in bioassay reports have been published
(7,8).

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                                     139
    The procedure for acute bioassay with fish is becoming
standardized.  Almost, without exception, present-day tests
incorporate:

    1) a series of test containers, each with a different but constant
concentration of the toxicant;

    2) a qroup of similar fish, usually ten, in each container;

    3) observations of fish mortality or other detrimental responses
during exposures which last between 1 day and about 1 week, often four
days;  and

    1) final results expressed as concentration tolerated by the
median or "average" fish.

    Beyond this, there are many other factors which make for good
practice, and these are summarized in the references cited above.  In
essence, the items cited below serve as a checklist of good bioassay
procedure.

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                                     140
Species

    A fish or other aquatic organism of local importance snould he
used in bioassays.  Preferably the test organism should be a game or
panfish, since they are generally among the more sensitive species.
Ability to duplicate an experiment would be enhanced if one used a
selected strain of fish (9).  A selected strain would also enable one
to determine the difference between toxicants more reliably and to
better determine the cause for variation in results due to different
test apparatus.  However,  a selected strain does account for different.
sensitivities due to the response to other stresses such as disease,
population stress, etc., which may be effecting a resident population.

    Differences between fish species in susceptibility to a toxicant
are generally less *-han miqht be expected - sometimes no greater than
the variability within a single species tested in different types of
water.  For example, trout and carp are about equally resistant to
some toxicants if tests continue for several days, giving the "coarse"
species time to react  (10,11).  On the other hand, larvae and
juveniles are most often more sensitive than adults and may be
preferred for the test procedure.  For projecting entire ecosystems,
criteria suitable for the selected test fish will often also be
suitable for other aquatic animals and plants.  Thera are exceptions
to this generalization.  For example, ccpper is guite damaging to
algae and molluscs, and insecticides are especially dangerous to

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                                     141
aquatic arthropods.  There are data to allow one to predict these
situations.  When they are expected, bioassays should also be run with
invertebrates and algae  (12).
    When possible, the bioassay should te conducted in waters obtained
from the water body of concern since characteristics of me water,
e.g., hardness, are included in the test results.  Work ot less
immediate application, for example, when attemptinq to predict the
toxicity of a new waste under various conditions, a selection of water
types should be tried.  *here is considerable merit in using both hard
and soft, synthetically prepared dilution waters (13)  or varying the
salinity of marine systems.  There is great danger in using "tap
water" for dilution, if it has been chlorinated. Even d ech lor i nation
procedures do not always remove the last traces of chlorine, and much
of the past toxicity research is accordingly suspect.

    Tap water can contain dangerous concentrations of copper, zinc or
lead from plumbing.

    Variations in other physical-chemical characteristics of water
such as temperature, organic content and pH commonly atfect toxicity
of pollutants.  Effects of five such environmental entities on the
lethal threshold of ammonia were illustrated a decade ago (13, 11).

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                                     142
    Water hardness is particularly important in toxicity of metals.



Hydrogen ion concentration is a most important modifying factor for



ammonia and cyanide.  Higher temperatures sometimes increase toxicity



of a pollutant, but recent work shows that phenol, hydrogen cyanide,



ammonia, and zinc may be more toxic at low temperatures (15).







    In static tests there should be two or three liters of water per



gram of fish, changed daily, or increased proportionally in volume for



the number of days of the test.  In continuous flow tests, the flow



must provide at least two or three liters of water per gram of fish



per day, and it must egual test-volume in five hours, giving 90



percent replacement in half a day or less.







Acclimation







    Acclimation of the test organism to the new environmental



situation encountered before the bioassay begins may have a marked



effect upon the outcome.  Abrupt changes of quality of holding water



should be avoided.  Time for acclimation of the organisms to the new



test conditions including temperature, should be as generous as



possible and dependent on life-span, at least two weeks is recommended



for fish.







Test Methods

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                                     143
    These must be adequately described when the results are given and



the easiest way is to follow one of the recommended bioassay



procedures in Table 1.







Controls







    Adequate and appropriate control tests must always be performed



(2).  Survival of control organisms is a minimum indication of the



quality of test organisms.  In addition, levels of survival and health



in holding tanks should be indicated and the conclusions recorded.

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                                     144

                               TABLE 1

          LITERATURE SOURCES FOP BIOASSAY AND BIOMONITORING
              PROCEDURES WITH VARIOUS AQUATIC ORGANISMS
Kind of       Type of
      §E	Response
                    Appropriate Situations
                   	for_use	
                              Reference
Fish and
Macroin-
vertebrates
96 hour lethal
concentration
To measure lethal toxicity
of a waste of known compostion,
to serve as a foundation
for extrapolating to pre-
sumably safe concentrations.
Fish and
Macroin-
vertebrates
Lethal threshold
concentration
For research applications
for document lethal thres-
holds.
 2,  5
Fish and
inverte-
brates
Incipient lethal
temperatures and
ultimate incipi-
ent lethal
temperatures
Research to determine
lethal temperature ranges
of a given species.
17,  18

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                                     145
                           TABLE 1 (CONT'D)
Kind of
              Type of
 Appropriate  Situations
	for_Use	
                                                                Reference
Fish
              Respiratory
              movements as
              acute sublethal
 Quick (1  day)  indication  of
 possible  sublethal  effects.
 Research  and  Monitoring.
19
Pish (iAe_.
fathead
minnows,
brook trout,
bluegill)
              Reprodution,
              growth, and
              survival
 Chronic tests  for  research
 on  "safe"  concentrations.
20-25
Daphnia
              Survival 6
              Reprodution
 Rapid completion  of  chronic
 tests for  testing special
 susceptibility  of crustaceans
25
Diatoms
              Survival, growth
              and reproduction
A  sensitive,  rapid, chronic
test  for  research, pre-
diction,  or monitoring*
26

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                                     146
                           TABLE 1  (CONT'D)
Kind of



Organism
              Type of
Appropriate Situations



      	for Use
                                                                Reference
Marine



Crustacean



Larvae



Mollusks
              Survival, growth



              and development



              through immature



              stages.
A sensitive, rapid, chronic



test, for research, pre-



diction, or monitoring*
27
*  Requires an operator with some specialized biological training.

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                                     147
                  Environment
    The supply of dilution wat>?r must be adequate to maintain constant
test conditions.  In both static and continuous-flow tests, a
sufficiently large volume of test water must be used, and it must be
replaced or replenished frequently.  This is to provide oxyqen tor the
organism, dilution of metabolic wastes, limit changes in temperature,
pH, etc., and to compensate for degradation, volatilization, intake
and sorption of the toxicant.  In static tests, there should be 2 or 3
liters oxygen per gram of fish, changed daily or else the volume
proportionally increased for the number of days of the test.  In
continuous-flow tests, the flow must: (a)  provide at least 2 or 3
liters oxygen per gram of fish per day, and  (h) flow must equal test-
volume in five hours or less, giving 903? replacement in half a day or
less.

    The problem of maintaining dissolved oxygen concentrations
suitable for aquatic life in the test chamber can be difficult.  The
suggestions above on test-volume and replacement times should provide
for adequate oxyqen in most cases.  However, with some wastes,
insufficient oxygen may be present in the test water because BOD or
COD may consume much of the available dissolved oxygen.  Aeration or
oxygenation may degrade or remove the test material.  Devices for
maintaining satisfactory dissolved oxygen in static tests have been
proposed and used with some degree of effectiveness, and have been
described (28).

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                                     148
    Periodic measurements of concentration of the toxicant should b
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                                     14P
time versus LCSO could then be constructed on loqarithroetic axes.  The
lethal threshold concentration could then be estimated in many cases
(3, 5, 6)  and this usually provides a more valid single number for
description'of acute toxicity than the arbitrary-time 96-hour LC50.

Test_Organis m s
    For important bodies of water, there is a strong case for testing
several kinds of aquatic organisms, not only fish.  A comparative
study of 20 pollutants on fish, snails, and diatoms found that no
single kind of organism was most sensitive in all situations  (10).
    The short-term bioassay method for fish can also be used  for many
of the larger invertebrate animals.  Few changes in method are
necessary except those of size, and a much greater volume of  test
water and/or rate of flow in relation to weight of th» animals, since
their metabolic rate is hiaher on a weight basis.
Metho^s^or_ysin2_Diatoms_as_fiioa^say__gr.ganisms_i.n_the Laboratory
    Algae and other microorganisms are uspful for bioassays because a
chronic test can actually be done in a week, since cycles of
reproduction and growth occur  in that time  (34).  It is important in
selecting one to remember that a kinetic  rather than a two-point
measure of growth rate is necessary to avoid errors introduced by lag
and recovery ohenomona commonly encountered.

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                                     150
    Short-term or acute toxicity tests do not indicate concentrations
of a potential toxicant that are harmless under conditions of long-
term exposure.  It is desirable, therefore, to multiply the 96-hour
values by factors, to estimate concentrations of the constituent in
question that is safe in the receiving water.  Such factors have been
called application factors.  An application factor is not a safety
factor since it makes no extra allowance of safety for unknown
factors.  It is merely a fractional or decimal factor applied to a
lethal or effective concentration in a short  term test, in order
to estimate the safe concentration as accurately as possible.

    Ideally, an application factor should be experimentally determined
for each pollutant.  To do this, it wculd first be necessary to
determine the lethal concentration of the constituent (s) in question
according to the bioassay procedures outlined above.  To obtain the
application factor it would be necessary to determine the truly "safe"
concentration of the same constituent(s)  on the same species by
thorough research on physiological, biochemical, and behavioral
effects, and by studying growth, reproduction, and production in the
laboratory and field.  The safe-to-lethal ratio obtained can then be
used in the future as an application factor, to work from measured
LCSO's of similar kinds of water, to predict the safe concentration.
Such bioassays should be repeated at least monthly.

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                                     151
    For example, if the 96 hour LC5C is 0.5 tng/1 and the concentration

of the constituent found to be safe is 0.01 mg/1, the ratio would be:



         Safe Concentration        0.01       1
         	   =    	   =  	  =  0.02
         96-hour LC50             0.50       50

    In this instance, the safe-to-lethal ratio is 0.02, which can be

used as an application factor in other situations.  Then, in a given

situation involving this constituent, the safe concentration in the

receiving stream would be found by multiplying the four-day LC50 by

0.02.



    This predictive procedure based on lethal concentrations is useful

since the safe level of many pollutants is not known, because of the

uncertainty about the toxicity of mixed pollutants because ot the

difference in sensitivity among fish and fish food organisms.  The

various considerations involved in developing application factors are

discussed in the literature (25).  Results of studies in which

continuous exposure was used reveal that the safe-to-lethal ratio

which permits spawning ranges over nearly two orders of magnitude (20-

21) .  It is recognized that exposure in most natural situations will

not be constant and that higher concentrations usually can be

tolerated for short periods.



    Lethal threshold concentrations, which may reguire more than 96-

hour exposure, may be beneficially used  to replace 96-hour LCSO's

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                                     152
in the above procedures, and there is a current trend to use such
threshold concentrations (2U).
Mixtures of Two or More Pollutants
    For the lethal action of a mixture, the t.oi-al toxicity may be
estimated by expressing the actual concentration of each toxicant as a
proportion of its lethal threshold concentration (= 96 hour LC^
usually)r then addinq together the resulting number for all the
toxicants.  If the total is 1.0 or greater, the mixture is predicted
to be lethal.

    The system of "adding up" different toxicants is based on the
premise that their lethal actions are simply additive, one with the
other.  This simple rule has been found to govern the combined lethal
action of many pairs and mixtures of quite dissimilar toxicants and is
true for such diverse toxicants as copper and ammonia: and zinc and
phenol in both laboratory  (36-38) and field studies (39).  The method
of addition is useful and reasonably accurate for predicting
thresholds of lethal effects in mixtures.  Also, there are cases where
the combined actions of two or more toxicants exceed the simple
additive effects of the individual toxicants.  This phenomenon is
commonly referred to as synergism.  No technique is known for
predicting increased lethality of mixtures of toxicants acting
synergistically.  Conversely there is evidence of mixtures of two or

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                                     153
more compounds havinq a lethal effect that is less than the single
additive effects of -t-.he individual toxicants.  This phenomenon is
commonly referred to as antagonism.  Techniques for predicting the
lethality of mixtures of antagonistic consitituents also are not
known.
    Sublethal or chronic effects of mixtures are of great importance
for water quality criteria.  Sublethal concentrations of different
toxicants should also be assumed to be additive in effect.   Here
again, it would be expected that for any giver toxicant there would be
gome low concentration which would have no deleterious etfect on an
organism, and would not contribute any sublethal toxicity to a
mixture.  Research is extremely scanty on this topic.

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                                     154
1.   Alderice, D. F. 1967.  "The detection and measurement
         of water pollution - biological assays".  Canadian
         Dept. of Fish.  Kept. No 9.  33-39


2.   Sprague, J. B. 1969.  Measurements of pollutant
         toxicity to fish.  I,  Bioassay methods for acute
         toxicity.  Water Res.  3_:   793-821


3.   Sprague, J. B.  1971.  Measurement of pollutant toxicity
         to fish.  III.  Sublethal effects and safe con-
         centrations.  Water Pes.  5:  245-266


U.   American Public Health Association, American Water
         Works Association, and Water Pollution Control
         Federation  (1971).  Standard Methods for Examination
         of Water and Wastewater.  13th ed.  American
         Public Health Association, Washington, D. C.
         87U p.


5.   Sprague, J. P., p.  p. Eicon and R. L. Saunclers, 1965.
         Sublethal copper - zinc pollution in a solnon river:
         A field and laboratory study, Air Water Pollution
         9_:  531-543

6.   Sprague, J. B.  1971.  Measurement of pollutant toxicity
         to fish.  II.   Utilizing and applying bioassay

-------
                                    155
         results.  Water Res.  U:  3-32







7.  Cairns, J.  1969.  Fish bioassays - reproducibility



         and rating.  Rev. Biol.  7:   7-12







8.  Cope, 0. B.  1861.  Standards for reporting fish



         toxicity tests.  Progr. Fish Cult.  23:  187-189







9.  Lennon, R. E.  1967.  Selected strains of fish as



         bioassay animals.  Progr. Fish Cult.  29:  129-132







10. Ball, I. R.  1967a.  The relative susceptibilities of



         some species of freshwater fish to poisons.



         I.  Ammonia.  Wat*»r Research 1,  767-775







11. Ball, I. R.  1967b.  The relative susceptibilities of



         some species of freshwater fish to poisons.



         II.  Zinc.  Water Research 1,   777-783







12. Patrick, R., J. Cairns and A. Scheier.  1968.



         The relative sensitivity of diatoms, snails and fish



         to 20 common constituents of industrial wastes.



                      Cult.  30,  137-140
13. Lloyd, R.  1961.  The toxicity of ammonia to  rainbow

-------
                                     156
         trout (Saimo aairdnerii^ Richardson) .   Water Waste



         Treat.  fl:   278-279







1U. Lloyd, R.  1961.  Effect of dissolved oxygen con-



         centrations on the toxicity of several poisons  to



         rainbow trout  (Saljrio gairdnerii Richardson) .



         J. Exp.  Biol.  38  447-455








15. U.K. Ministry of Technology.   1969.  Some effects of



         pollution on fish.  II.   Notes on Water Pollution



         NO. U5:    5.








16. Brown, V. M.   1968.   The calculations of the acute



         toxicity of mixtures of poisons to  rainbow trout.



         Water Res.  2:   723-733








17. Fry, F. E. J.  19U7.  Effects  of the environment on



         animal activity,  u. of Toronto studies, Biol.



         Serv.  No. 55.  Pub.  Ont. Fish Pes. Board No.  68.'



         62.








18. Brett, J. P.   1952.  Temperature tolerance  of young



         Pacific salmon, genus  Oncorhynchus J. Fish.



         Res. Board Can.  9:  265-323

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                                     157
19. Schamhurg F. D., T. E. Howard, C.  C.  Walden   1967.



         A method to evaluate the effects of  water pollutants



         on fish resperat.ion.   Water Res.  1:  731-737







20. Mount, D. I.  1968.  Chronic toxicity of  copper to



         fathead minows  (Pi.me2ha.ies  promelas, Rafinesquo) .



         Water Res.  2:  215-223







21. Mount, D. I. and C. E. Stephen   1967.  A  method of



         establishing acceptable toxicant limits for fish:



         malathion and butoxyethanol ester of 2, 4-D.



         Trans. Amer. Fish Soc.  9ji:   185-193







22. Brungs, W. A.  1969.  Chronic toxicity of zinc to the



    fathead minnow, PimeEhalas  B!2me,l.a.§ Rafinesque.



    Trans. Amer. Fish Soc.   98:  272-279







23. McKim, J. M. and D. A. Benoit   1971.   Effects of



         long-term exposures to copper-on survival, growth,



         and reproduction of brook  trout. (Salvelinus fontinalis)



         J. Fish. Res. Board Can.   28:  b"55-CG2







2«». Eaton, J. G,  1970.  Chronic malathion toxicity to



         the bluegill (Leponms  SiaciSClJiUSS' Rafinesque)



         Water Res.  U:  673-G84

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                                     158
25. Anderson, B. G.  1950.  The apparent thresholds of
         toxioity to Daphnia magria for chloride of various
         metals when added to Lake Erie waters.
         Trans, Amer. Fish Soc.  78:  96-113

26. Patrick, R.  196R.   Standard methods for tests for
         evaluating inhibitory toxicity of industrial waste
         waters in American Society for Testir.q and Materials
         Book of Standards, part 23:  Industrial Water; Atmospheric
         analysis  (American Soc.-for Testing and Materials,
         Philadelphia,  Pa.)  pp 657-665

27. Woelke, c. E.  1967.  Measurements of water quality
         criteria with Pacific oyster embryo hioassay.
         Amer. Soc. Test. Mater. Spec. Tech. Publ no. U17:   112-120

28. Doudoroff, P. , B.  G. Anderson, G. E. Burdick, P.S.  Galtsoft,
         W. B. Hart, R. Patrick, E. R. Strong, E. W. Surber; and
         M. W. VanHorn.  1951..  Bioassay methods for the evaluation
         of accute toxicity of industrial wastes to fish.  Sewaye
         Indust. Wastes.  23;  1380-1397

29. Brunqs, W. and D. I. Mount, 1967.  A device for continuous treatment
         of fish in holding chambers.  Trans. Am. Fish. Soc.

-------
                                     159
         96: 55-57








30. Stark, G. T. C.  1967.  An automatic dosir.q aoparatus made



         with standard laboratory ware.  Lab. Pract. 16: 594-595








31. Mount, D. I. and Warner. R. E.  1965.  A serial-dilution



         apparatus for continuous delivery of various conentrations



         of materials in water.  U.S. Publ.Health. Service,



         Pub no. 999-WP-23, 16pp.








32. Mount, D. I. and Brunqs, W. A.  1967.  A simplified



         dosing apparatus for fish toxicology studies.



         Water Research 1: 21-29.








33. Litchfield, J. T. and F. Wilcoxon,  1949.  A simplified method of



         evaluating does-effect experiments.



         J. Pharmac.  Exper. Therapeutics,  ^6:  99-113








3«». Patrick, Ruth,  196U.  Tentative method of test for



         evaluating ingihitory toxicity of industrial waste



         waters.  American Soc. of Testino and Materials.



         pp 517-525








35. Henderson, C.  1957.  Applicaiton factors to be applied



         to bioassays for the safe disposal of toxic wastes.

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                                     160
         Biological Problems in water Pollution.  Trans, of 195h



         Seminar U.S. Public Health Servic_, P.A. Taft Sanit. Enynq.



         Center, Tech. Dept.  W60-3 : 31-37








36. Herbert, D. W.  M. and Van Dyke, Jennifer M.  196U.  The



         toxicity to fish of mixtures of poisons-IT.  copper-



         ammonia and zinc-phenol mistures.  Ann. Anpl. Biol.



         53 : 415-5121








37. Jordan, D. H. M. and Lloyd R.  196U.  The resistance



         of rainbow trout (Sa^mo Gairdnerii, Richardson) and roach,



         (Rutilus rutilus L.) to alkaline solutions.



         Tnt. j. Air Wat. Pollut. 8 : 405-U09








38. Brown,  V. M. Jordan, D.  H. M. and Tiller, B. A.  1969.



         The acute toxicity to rainbow trout, of fluctuating



         concentrations and mistures of ammonia, phenol and



         zinc.  J.Fish Biol. 1:1-9








39. Herbert. D. W.  M.  1965.  Pollution and fisheries.



         In Ecology and the Industrial society, 5th Symposium



         British Ecological Society,  pp 173-195.  Blackwell



         Scientific, Oxford.








40. Lloyd,  R. and Orr, Lydia, D.  1969.  The diuretic

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                           161
response by rainbow trout, to sub-lethal concentrations



of ammonia.  Water Research 3;  335-3UU.

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                                    162
    B.   Classification of Waters 30U(a)  (2) C




         Classification of waters, depending upon the suitability for
                      •
the protection of human health and for the propagation ot fish and

aquatic life is categorized as follows:




         1.    Protection^gf^Human Health




              a.  Acceptable:  Waters which are suitable

for contact recreation.




              b.  Treatable  (Acceptable with treatment):

Waters which, although not suitable for contact recreation,

are acceptable for drinking water supply intake and can be

treated to acceptable potable water meeting human health

standards.




              c.  Unacceptable:  Waters which contain

substances such as hydrocarbons  (grease, oils)  and hazardous

substances which cannot be removed and thereby render the

water unsafe for recreation or drinking water intake.

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                                    163
         2.   Protection of Fish and Aquatic Life







              a.  Acceptable:  Waters which are suitable



for the propagation of the most important sensitive species



of aquatic biot.a.







              b.  Unacceptable:  Waters that contain



concentrations of pollutants that, will not sustain "the most



important sensitive species" of aquatic biota.

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                                     164
V.  Identification  of Pollutants Suitable  for Maximum Daily Loads



    30U (a) (2)D







    All pollutants  described by Volume I,  Water  Quality Criteria, are



EStentially  suitable for maximum daily load  restriction; however, the



existence  of Water  Quality Standards is  a  prerequisite tor making this



determination.   Only those pollutants which  have a  speciiic limitincr



value  in the standards or those pollutants whose effects are



specifically limited in the standards are  suitable  for maximum daily



load.
«U.S. COVBRNMENT PRINTING OFFICEM971 54b-3i.VI4* 1-1

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