UNITED STATES STF1EL,  LORAIN,  OHIO,  WORKS,  BLACK- RIVER SURVEY:

ANALYSIS FOR HEXANE ORGANIC EXTRACTA3LES AND FOLYNUCLEAR AKCMATIC HYDROCARBONS
                    Herbert J.  Brass,  Supervisory Chemist
                          Walter C.  Elbert, Chemist
                           Mary Ann Feige,  Chemist
                          Edward M.  Click,  Chemist
                         Arthur W.  Lington, Chemist
                       Organic Chemistry Laboratories
              National Field Investigations Center - Cincinnati
                  Office of Enforcement and General Counsel
                    U. S. Environmental Protection Agency
                              5555 Ridge Avenue
                           Cincinnati,  Ohio 1*5268

                                OCTOBER  197U

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                         TABLE OF CONTENTS


                                                       Page Numbei

SUMMARY                                                    1

INTRODUCTION                                               3

EXPERIMENTAL                                               5

    A.  HEXANE EXTPACTA3LES (OIL AND GREASE)               5

    B.  LIQUID CHRCMATOGRAFHIC FRACTIONATION               6

    C.  GAS CHROMATCGPAPHY                                10

    D.  GAS CHROMATCGRAPHY/MASS SPECTROMETRY (GC/MS)      12

RESULTS                                                   15

    A.  WATER SAMPLES                                     15

    B.  HEXANE EXTRACTABLES (OIL AND GREASE)              17

    C.  POLYNUCLEAR AROMATIC HYDROCARBON ANALYSIS IN
        SEDIMENT SAMPLES                                  20

        1.  Liquid Chromatographic Separations
            of PAH            '                           20

        2.  Gas Chromatographic Analysis                  25

        3.  Gas Chromatographic/Mass Spectrometric
            (GC/MS) Analysis          "                    33

DISCUSSION                                                6k

    A.  SURVEY FINDINGS                                   6k-

    B.  OTHER COMMENTS                                    66

REFERENCES                                                6?

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                              SUMMARY




     Three water and twelve sediment samples,  collected at the U.  S.




Steel, Lorain, Ohio, works, were analyzed as part of an extensive




biological and chemical survey of the Black River in the vicinity  of




the U. S. Steel site.  Two water samples, taken from blast furnace




effluents, were free of contamination.  The coke sewer (002)  sample




did show the presence of small amounts of organic compounds,  possibly




paraffins, cycloparaffins, and/or olefins.  The concentration of any




individual component was less than 20 ppm (wt/v).  Small amounts




(<1 ppm) of polynuclear aromatic hydrocarbons (PAH) may have  been




present in the coke sewer sample.  Water sampleswere analyzed by gas




chromatography/mass spectrometry (GC/MS).




     The sediment samples were extracted with ji-hexane to determine




organic solvent extractables - "Oil and Grease".  Values ranged from




810 to 26,700 mg/kg based on dried sample weight.  Highest values  were




at sampling sites in close proximity to the plant.




     PAH were determined in sediment samples by gas chromatography (GC)




and GG/MS.  Liquid chromatography (LC). was used to separate PAH from other




classes of organic  compounds in hexarie extracted  residues.  Quantitation of




ten PAH in eight samples was then made by GC.   These PAH  represented an en-




velope of polynuclears  of  increasing  ring complexity ranging  from phenan-




threne and anthracene  (molecular weight  178) through two  dibenzanthracenes

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(MW 278). Several of the PAH quantitated are known carcinogens.  Values of




benz[a]- plus benz[e]pyrene ranged from 0.91 nig/kg (sample 19^66) to




19 mg/kg (sample 19^72).  Values for dibenzanthracenes ranged from 0.37




(19U671 to 12 mg/kg (19472).  Sample 19^72, collected just downstream




from plant outfall 002, contained the largest concentrations of PAH;




this sample was examined in detail "by GC/MS.  A "broad spectrum of PAH




was seen to be present: these included unsubstituted as well as methyl




substituted polynuclears.   Specialized GC/MS data system techniques




were used to determine the presence of PAH.  Isotope abundances were




used to confirm molecular formulas.

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                           INTRODUCTION



     At the request of EFA Region V,  NFIC-Cincinnati's Organic Chemistry




Laboratories vere ad-ted to analyze twelve sediment and three water samples, •



collected by NFIC-Cincinnati personnel, at the U. S. Steel, Lorain, Ohio,



works; this plant discharges into the Black River.  This request is part



of an extensive biolo-rical and chemical survey of the Black River in the



vicinity of the U. S. Steel site in conjunction with EPA's pending court



case against U. S. Steel.  The sediment samples were to be analyzed for



compounds extractable into an organic solvent; this procedure is better



known as an Oil and Grease Determination.  In addition the sediments




were to be scanned for Polynuclear Aromatic Hydrocarbons (FAH).  Certain



PAH, for example benz[a]pyrene, are known to be powerful carcinogens.  The



three water samples were to be analyzed for the presence of organic com-



pounds and in particular PAH.  Chain of custody procedures were followed



in handling the samples.




     The request for PAH analysis arose out of the survey, conducted by



Mr. Allen Lucas of these laboratories,  in which substantial concentrations



of benz[a]pyrene (BaP) were found to be present in sediment samples collected



at a U. S. Steel - Calumet site.  Dr. Paul Clifford, who analyzed these



samples, reported concentrations of BaP ranging from 1 to 380 mg/kg (dry



sample weight) using gas chromatography/mass spectrometry (GC/MS) and



fluorescence spectroscopy as analytical tools.



     The approach taken in our laboratory to analyze for PAH in the Lorain



sediment survey was different than in the Calumet case.  In the latter

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instance FAji v.-ere found as part of a general scan for organic compounds;




phenanthrene, anthracene, and EaP were then selected as measures of PAH




and carcinogenic content.  In the Lorain case it was decided to analyze




the samples for a broad spectrum of PAH from phenanthrene and anthracene




(KW=178) to the dibenzanthracenes (MW= 2?8).  It was felt that it vould




be of greater value scientifically to examine selected sediments for many




individual FAH than all samples for, say, EaP.  In fact a detailed PAH




scan was obtained for eight of the twelve Lorain sediment samples.

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                           EXPERIMENTAL


A.  HEXAiYE EXTRACTA^LES (OTL Ai-JD GREASE)


     The method used for hexane organic extractables is essentially the

                                                   2
same as outlined in tr.e Standard Methods nrocedure.



     Samples were weighed into individual wide mouth porcelain crucibles.  Before


removal each sample was_agitated in its original container to assure'adequate


mixing.  A control was prepared.  It contained 25 g of silica gel and 25 ml


of water.  Samples and the control, ca 50 g T"ret weight, v/ere air dried at


30°C overnight in the individual crucibles.  They were then crushed


to fine particles and allowed to dry an additional two hours.  Each residue


was placed in a soxhlet thimble and extracted in individual soxhlet apparat-


uses into "distilled-in-glass" n-hexane for 10 hours.  Hexane was evaporated


to 15 ml by allowing a stream of nitrogen to flow over the solvent in the


extraction flasks.which were heated by a steam bath held at 80°C.  The re-


maining sample and solvent from each flask were transferred to appropriate


size vials.  The extraction flasks were washed with small portions of n-hexane


which were transferred to the vials.  No apparent residue remained in the ex-


traction -flasks.  A stream of nitrogen was allowed to flow over each sample


vial, at room temperature, until the hexane solvent had evaporated.  Checks


were made to be certain that samples had reached a constant weight indi-


cating complete hexane loss.  Oil and grease is reported as mg, organic


residue per kg, dry sample.   It should be noted, that in all procedures,


attempts were made to keep sample exposure to light in the laboratory to a


minimum to prevent decomposition of compounds to be analyzed for in the residue.

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3.  LIQUID C?I?.CMA7C'J?--.?r:IC F?ACTICNATTO:i




     Polynuclear aro-acic hydrocarbons were separated 'from other classes




of organic compounds in the hexane extractable residues by a liquid




chromatographic ' LC' procedure given by Hoffmann and Wynder.     In order




to determine recovery of FAH  from  the  LC  column  an  experiment  was




performed.




     Florisil, oO/ICO :nesh, was heated for an hour at 110°C and cooled




in a dessicator.  Forty grams of the activated Florisil was weighed and




placed in a column chroraatography tube whose dimensions were hO X 2 cm




(id).  The volume occupied by the Florisil was 20.5 X 2 cm (id).  Aluminum




foil was placed around the column to prevent any possible photochemical




reactions.  With the stopcock in an open position, hexane was poured into




the column.  (it is imperative that the solid packing be uniform and with-




out air bubbles).   The initial hexane elutant was discarded.  The volume




of eluting solvent passing through the column was determined to be 37 ml.




Five ml of a 1 mg/nl standard of BaP in cyclohexane was applied and ad-




sorbed onto the column.




     The first eluting solvent was n-hexane - 200 ml.  The flow rate was




375 ml/hour.  Twenty-five ml fractions were collected and viewed under




ultraviolet (UV) light (long wavelength - 365 nm) and observed for fluor-




esence.  No fluorescence was observed'for all eight fractions.




     The second eluting solvent was an 8:1 hexane/benzene (V/V) solution.




The volume of this solution to be applied was determined experimentally.




A critical variable which affects the total volume of the solvent used

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is the flow rate (i.e., slower flow rates require less eluting volume).



A flow rate of 3^0 ml/hour was established.  Fifty ml' fractions were



collected in 100 ~1 reagent bottles.  Each fraction collected was covered



with aluminum foil and viewed under UV (365 nml light.  The total volume



after which no further fluorescence was observed was 700 ml.  The frac-



tions were pooled and the total volume (TOO ml) was concentrated in a



1 liter round bottom flask by use of a Euchi ROTAVAPOR-R.  After the sol-



vent was evaporated to approximately 30 ml, the concentrate was trans- .



ferred to a 250 ml wide-mouth flask.  The original container was washed



with 8:1 hexane/benzene and the washings transferred to the 250 ml flask.



Under a steam bath (at c_a 70°C) and a gentle stream of nitrogen, the volume



was further reduced to 2-5 ml.  This solution was transferred to a 15 ml



centrifuge tube and the 250 ml flask washed with 8:1 hexane/benzene;



washings were added to the centrifuge tube and this volume was reduced to



ca 2 ml and stored in a refrigerator until ready for analysis by GC.



     Samples were chromatographed in a fashion similar to that in the BaP



recovery experiment.  The Scheme (Page 8} outlines the procedures used.  The



samples (i.e., hexane extractable organic residues) were dissolved in 6 ml



n-hexane and applied to:Florisil columns.  n-Hexane was the first solvent;



it was employed to elute non-polar paraffinic type material.  Depending on
                                     /
                                    /

the sample, 250 to 1000 ml of n-hexane were used. The n-hexane fraction



is defined as fraction A.  For one sample, number 19U?0, where a duplicate



oil and grease determination was made, ten 100 ml fractions were collected

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           FLOWCHART OF TYPICAL  SAMPLE  WORKUP
                                                       fR~~l
                                                       I LJi  I
                                                       \\i || II \ i-ifi^mlf t
                 8=1
           HEXANE/
           BENZENE
{SAMPLE  IN

|e Ml ti*-HEXANE
 FLORISIL
ICoL. CHROM
 N-HEXANE ML
'25O-1OOO
                   1-3
             LITER
B
CONCEN.
                            ML
B  P
N) T RQMETHANF
                                     EXTRACTION
                              HEXANE

                                LAYER
                             EVAP. DRYNESS
                             PVEOISSOLVE
                             N—HEXANE
                                                       GC
                                                      GC/MS
                        SCHEME

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in 125 ml Erlersneyer flasks.  No fluorescence was observed indicating no



"breakthrough of PAH.  Fractions 2,  U,  6, and 10 were concentrated to ca



5 ml on a steam bath under a stream of nitrogen.  The concentrates of



these fractions were examined by gas chromatography.



     After ri-hexane, an 8:1 n-hexane/benzene (V/V) solution was used to



elute fraction B which contains the PAH.  As fractions were collected



they were checked for fluorescence.  Certain of the samples less concen-



trated in PAH, particularly in higher molecular weight PAH, displayed no



fluorescence after c_a one liter of solvent had been eluted from the column.



For these samples elution was terminated when fluorescence was no longer



observed.  Other samples, for example 19^72, displayed fluorescence after



elution by 3«5 liters of solvent.  For this sample eluate from 0-2500 ml



and 2500-3500 ml were separated.  •



     Fraction B was concentrated to 10 ml by previously described methods.



It was then extracted four times with 10 ml portions of nitromethane (NM). '



The NM extracts (fraction B_T in Scheme) were pooled and evaporated to



dryness and redissolved in n-hexane—1 to 5 ml depending on the sample.

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c.  GAS CHROMATC:RAPHY

     A gas chrorcatcgraphic (GC) method was employe^ to analyze for specific


PAH, based oh the system of Lao et al.   A 10 ft X 1/8 in stainless steel

column was employed which was packed with 6% Dexsil 300 on 80/100 mesh Supel-

coport.  A similar column packed with 3$ Dexsil 300 was equally efficient


and eluted compounds in a shorter time.  A Varian lUOO GC equipped with a

flame ionization detector (FID) and a Hewlett Packard Model 7100B, 1 MV

recorder, were used.  Injector temperature was 225°C and detector tempera-


ture 325°C.  After injection the column-was held at l65°C for two minutes

then programmed at U°C per minute to 295°C and held until the

dibenzanthracenes had eluted.  Total analysis time for the 6$ arid 3% Dex-

sil columns were ca 120 and 80 minutes, respectively.  The carrier gas

was heliuin and its flow rate 30 ml/min.  The cylinder pressure regulator
                                                                         V
was set to deliver 80 psig to"minimize pressure drop accross  the  column


during the temperature program.  Extreme care had to be taken to  prevent


helium leaks in the carrier gas line.  A Gow-Mac Model 21-110 Leak Detector

was useful in detecting leaks in gas  lines,

     Hamilton 10 nl syringes were employed for injection of standards and

samples onto the GC column.  Standards (1000 ppm) were prepared in Burdick


and Jackson distilled-in-glass ri-hexane using PAH obtained from commercial

sources.  Perylene was dissolved in benzene due to its low solubility in

n-hexane.  For quantitation, a standard mixture of ten PAH (see Results)

was prepared in ri-hexane.  The standard mixture consisted of phenanthrene,
                                10

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anthracene, fluorar.thene, pyrene, benz[a]anthracene, benzfajpyrene,



benz[e]pyrene, perylene, dibenz[a,h]anthracene, and dibenz[a,c]anthracene.



The concentration of each PAH in the standard was 50 ppra (wt/v) except for



dibenz[a,h]anthracene v/hich was 25 ppra.  Concentrations of PAH in the




samples were calculated by comparison with the standard mixture.  Peak



heights were used to compare the earlier eluting PAH (through benz[a]anthra-



cene); peak areas were used for quantitations of BaP and BeP and later eluting




compounds.  In calculating concentrations of PAH components, corrections



were applied for volumes of samples and standards analyzed and for total



sample volumes.  For comparison, the standard mixture was analyzed prior



to sample analysis.  When more than one sample was to be analyzed, the



standard was run after every 2-3 samples.



     In analyzing the paraffinic fraction A, a Perkin-Elmer Model 900 gas



chromatograph equipped with a dual FID system and a Honeywell potentiometric



recorder was employed.  The f>C was operated in the dual differential mode.



Two 6 ft X 2 mm (id) glass columns packed with 3% OV-1 on 60/80 mesh



Supelcoport were employed.  Injector and detector temperatures were 220°



and 320°C, respectively.  The column was held at 80°C for five minutes after



injection and then programmed at 10°C/minute to 220°C.  The temperature was



then held for ca 10 minutes.
                                   11

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D.  GAS CHHOMATOCRA~-Y/:.IASS S?ECTRO>-ETRY (GC/MS1


     Analysis of standards and samples by OC/MS employed a Finnigan Model


1015 quadrupole GC/MS equipped with a System Industries Model 250 data


system.  Inasmuch as the C-C on the Finnigan unit is a modified Varian lUOO


instrument, columns could be interchanged with the GC possessing the FID.


     The mass spectrometer (MS) was scanned by applying mass set voltages


to the quadrupole rods from a 15-bit digital-to-analog converter.  At a


given mass set voltage, only ions of a specific mass-to-charge ratio pass

                                                                 7
through the quadrupcle field to the electron multiplier detector.   The high

                                                              _y
mass range of the MS was used; the preamplifier setting was 10   amp/volt and


2600-2800 volts were applied to the electron multiplier.


     Direct aqueous injection of water samples by GC/MS was accomplished

                                Q
using the method of Budde et al.   A 6 ft X 2 mm id, glass column was packed


with Chroraosorb 101 and conditioned at 270°C for two days.  Five ul of


sample was injected and the column oven held at 80°C for two minutes and


then programmed at h° or 8°C/minute to 220°C.  The final temperature was


held so that the total analysis time was 35-UO minutes.  The MS ionizer .and


data system were turned on after water had eluted from the GC column and been


pumped from the ion source.  It was allowed to scan from 33-200 amu.


     Water samples were also extracted and the extracts examined by GC/MS.


One liter of sample was extracted twice with distilled-in-glass methylene chlor-


ide.  The extracts were combined and dried by passage through two inches of
                                 12

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prewashed (with methylene chloride)  anhydrous sodium sulfate-in a glass col-


umn.  After the combined extracts had been dried, 100 ml of distilled-in-


glass acetone was poured through the column. The extract plus acetone was evap-


orated to 5 mi employing a Kuderna-Danish apparatus.  In this way the


methylene chloride (BP 39.8°C) was removed, leaving the sample dissolved in


acetone (BP 56.1°C).


     A 6 ft X 2 mm (id) glass column packed with 1.5$ OV-17 + 1.95$ QF-1


on Supelcoport, 100/120 mesh, was used to analyze extracted water samples.


It was conditioned at  250°C for two days prior to analysis.  Two to four ul


of sample was injected onto the column and .the MS .was "turned on" after  the


solvent, acetone, had been pumped from the ion source, ca 1 to 2 minutes,


and allowed to scan from 33-^50 amu.


     For sediment samples the 3$ Dexsil column described in the section on


gas chromatography was used.  At the high temperatures needed for these


analyses (ca 3CO°C) GS/MS separator and transfer lines had to be adjusted


to 280°C and 230°C, respectively.


     The System 250 data system has certain special features which were

                             p.
applicable to these analyses.7  These include choosing mass set voltages


so that one can acquire data for selected ions of interest.  This technique


has the advantage of allowing the'detection of compounds at 10-100 times

                                    /

lower concentrations than by normal''scanning techniques.  Another feature


is being able to search the reconstructed gas chromatogram (RGC) for a


limited number of ions.  The RGC is a plot of ion current, normalized to 100$


for the spectrum yielding the highest'ion current against spectrum number.
                               13

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The limited mass search allows the operator to ignore background and




search for contributions to the ion current by selected ions of interest



as a function cf spectrum number.
                                1U

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                              RESULTS




A.  WATER SA..-r?LZS




     Three water samples were received.  The first was taken at the 1, 2,




5 blast furnace (east clarifier) ,  the second, at the 3, U blast furnace (west




clarifier), and the third at the 002 discharge coke plant sewer.  The samples




were scanned for organic compounds, using the laboratory's gas chromatograph/




mass spectrometer (3C/MSX- unit.  Two approaches were taken.  First, a direct




aqueous injection of sample was made into the GC/MS.  This allows the




determination of lev.- molecular weight, volatile, organic compounds of




varying types.  The second procedure involved extraction of the sample into




raethylene chloride,  addition of acetone, evaporation, and concentration (see




Experimental Section).




     The water samples  were found to be essentially free of contamination.




The direct aqueous injection technique showed that the two blast furnace




effluents were free of lower molecular weight compounds though one of them




probably contained a small amount  of acetone.  The coke plant sewer sample




did show small quantities of organic compounds; the concentration of any




individual component was likely less than 20 ppm (wt/v).  While individual




components were not identified, two or three classes of compounds were



found possibly to be present; these are paraffins, cycloparaffins, and olefins.




These classes were searched for by allowing the computer to scan the GC/MS




reconstructed gas chromatograph for the presence of particular ions -




m/e U3 and 57 for paraffins and m/e 55 and 69 for cycloparaffins and




olefins.
                                15

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     Few organic compounds were present in samples extracted with methylene



chloride, even though the extraction procedures concentrated the samples



some 200 fold.  Blast furnace effluents showed no detectable compounds,



while the coke plant sewer again showed small GC/MS peaks.  For this



sample the GC/MS was set to acquire data solely for ions corresponding to



base peaks, which are also the molecular ions, of certain PAH.  The ions



were m/e 128 (naphthalene), 166 (fluorene), 1?8 (phenanthrene and anthracene),



202 (fluoranthene and pyrene), 228 (benz[a]anthracene and chrysene), and



252 (benz[a]pyrene and benz[e]pyrene).



     GC/MS peaks were obtained, for the coke sewer' extracted sample cor-



responding to m/e 166, 1?8, and 202 with 1?8 being the largest.  These data



indicate the presence of low concentrations (
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B.  HEXANE EXTRACT.-^ LES (OIL AND GREASE)




     Table I presents data for the n-hexane extraction of organic compounds-



from the twelve sedir.ent samples collected at the Lorain site.  Table II



gives a description of the sampling sites.  Values ranged from 810 to 26,700



mg/kg based on the -.-.-eight of samples dried overnight at 30°C.  A blank



containing 25 g of silica gel and 25 ml of water yielded no significant



oil and grease.  Five duplicate determinations were made and yielded excel-



lent reproducability.
                                17

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                                                 TABLE I

                                          "OIL AND GREASE" DATA
                                         Lorain Sediment Samples
                                                                   Weight Organic
Sample
Number
Blank *
1Q1*65
19'i66
19*167
19li67(A)
19U68 '
191i69
i9i(69(A)
191*70
191*70 (A)
19^71
19^72
191*72 (A)
19^73
191171*
19'*7U(A)
19^75
19W6
Wet Sample
V/t . , Grams

70.1*6
50.83
53-02
52.98
51.96
60. 1*3
57.82
52.67
1*9.70
1*9-57
55.53
55-27
1*9.08
61*. 79
62.3U
50.83
73-05
Dry Sample
Wt., Grams

30.79
25.62
28.8U
28.85
33.35
3**.53
33.30
32.69
30.83
20.1*6
36.71*
36.09
29.08
39-26
36.31
31.26
37.25
Percent
Volatiles

56.1*
'19.6
1*5-6
1*5.6
35.9
1*2.9
1*2 ".5
38.0
38.0
58.8
33.9
3^.8
39-6
39-5
1*1.8
38.7
1*9.1
Residues ,
Grams
0.0031
0.0258
0.0595
0.1170
0.0956
0.051*3
0.3683
0.3319
• 0.3080
0.3007
0.2889
0.9539
0.9919
0.5297
0.9758
0.9560
0.2065
0.2061
Oil and Grease, pprn
mg/kg Dry Semple

837
2,320
if ,050
3,310
1,620
10,700
9,960
9,lf20
9,750
1'* , 100
25,900
27,1)80
18,200
2l|,800
26,1*10
6,580
5,530
* Blank contained 25 g. Silica Gel and 25 ml. water

A - Duplicate determinations
                                                   18

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                                  TABLE   II

                 DESCP.IPT:::: c? BLACK RIVIR SAKPLIIIG STATIONSa
Sample
Iromber
19U65
19U66-
19U67
19^68
19^69
19^70
Sample
Station
1
2
3B
UB
5C
6B
?o
-
-
0.
1.
2.
2.
ile
ir.t


C
0
~>
75
Site Description
Lake water Treatment Intake
Mouth of Harbor: Lake Erie -Black


River
Center of Black River (River Mouth)
Center of River
Right Hand Side of River, Looking
Middle of Turning Basin Opposite

Upstream
Outfalls
                                   003 and

19U71      7B          2.9         Head of River Turning'Basin - End of
                                   Commercial Navigation

19^72      8B          3.U         ca 500 ft. Downstream from Outfall 002

19U73      9B          k.O         Near Plant Water Intake

19V7U      10          5.0         ca 0.2 mi. Downstream from Outfall 001

19^75      11          0.2         0.2 mi Upstream on French Creek

19^76      12          5.3         ca 0.2 mi. Upstream from Outfall 001
a
 'All samples taken from the surface of the sediment layer.
                                      19

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C.  POLYNUCLEAR AROMATIC HYDROCARBON ANALYSIS IN SEDIMENT SAMPLES



     The organic residues obtained in the Oil and Grease determinations



were used to analyze for PAH.  These residues are complex mixtures of



organic compounds of varying classes and a method of separating the PAH



(fraction) was necessary.  The largest body of information on the separa-



tion and analysis of PAH, other than in the air pollution field, lies in



the tobacco smoke chemistry area.  PAH determinations in organic residues



obtained from sediments should be.similar to determinations of PAH in



tobacco tars.  Separation of PAH was affected by the method of Wynder and



Hoffmann.     Several gas chromatographic (GC)  ~.  .and gas chromatographic/



mass spectrometric (GC/MS) " methods of measuring PAH are available.  Tech-



niques for GC and GC/MS analysis advanced by Lao et al.  appeared to be



most applicable and were employed.



     1.  Liquid Chromatographic Separations of PAH



         PAH were separated by the liquid chromatographic (LC) procedures



outlined in the Scheme and detailed in the Experimental Section.  Paraffinic



type compounds are eluted from an LC Florisil Column by n-hexane (fraction A)



The next solvent is an 8:1 hexane/benzene solution which elutes the PAH



(fraction B).  B is concentrated and PAH extracted into nitromethane



(fraction Bj^) to separate the PAH from-other organic compounds that are



eluted in fraction B.




     Prior to analysis of the sediment organic residues an experiment was



devised to measure PAH recovery from the LC column.  Five ml of a 1 rag/ml



(1000 ppm) benz[a]pyrene (BaP) solution' was applied to a column and eluted
                                20

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as in the Scherae except that the nitromethane (KM)  extraction was omitted.


The recovery of EaF was 97$.


     It was deterained that 1 liter of n-hexane was likely a sufficient


amount of solvent to elute fraction A.  For sample  19^70 1 liter of


n-hexane was eluted from the LC column with ten 100 ml fractions "being


collected.  Fractions 2, h, 6 and 10 were concentrated and examined "by

gas chromatography.using OV-1 as the liquid phase;  OV-1 is efficient for


non-polar materials.  Fractions 2 and U contained a broad envelope of


paraffinic type material which eluted from the OV-1 column as temperature


was programmed.  Fraction 6 was free of GC peaks until a high column


temperature was reached, ca 200°C.  Reasonably, this indicates high mol-


ecular weight compounds which were late eluters from the LC column.


Fraction 10 contained no GC peaks, thus signifying  a complete separation


of total fraction A.


    .Table III lists the volumes of n-hexane used to elute fraction A in


each sample.  Prior to GC analysis of the n-hexane  cuts of sample 19^70,


it was felt that 200-UOO ml of n-hexane would be sufficient to elute a


total fraction A.  Clearly this is not the case. Fortunately, however, any


carryover of paraffinic material into Fraction B did not appear to adversely


affect the final analysis; the nitromethane extraction apparently is almost
                                      /
exclusive for PAH, leaving behind other organic compounds in the hexane


layer.  For example GC/MS analysis of sample 19U?2  (see below) revealed


only PAH; few other types of compounds were present.  An exception was
                                21

-------
            VOLL7-ES  C? ^-HEXAiTE USED TO ELUTE FRACTION A


 Sample  'lumber                           Volume n-Kexane, ml

      19^66                                      1000

      19U67                                       ^00

      19^69                                       Uoo

      19^70                                       Uoo

      19U71                                      1000

      19U72                                       200

      19U7U                                       200

      19^76                                      1000

      19^70 (Duplicate)3"                         1000
aA second "Oil and Grease" determination was made on  this  sample
 see Table I.
                                 22

-------
sample igUyk where an exceedingly high paraffin content (see explanation



below) did interfere somewhat with PAH analysis.  Elution of fraction A



with greater than 200 nl of ji-hexane was obviously necessary for this




sample.



     No fluorescence was observed in the 1 liter of n-hexane eluted for



sample 19^7U.  However, a low level of fluorescence was detected for



samples 19^66 and 19^76.  The possibility does exist that small amounts



of low molecular weight PAH were lost.



   .  One to three liters of 8:1 hexane/benzene was used to elute Fraction B.



Table IV gives .the volumes of solvent used.  Elution of samples 19^66 and



19U76 was terminated at 1600 and 1250 ml respectively since fluorescence



was no longer observed in solvent passing through the column.  For sample



19^72 2.5 liters of solvent were eluted; this was followed by elution with



an additional liter which was separated from the initial 2.5 liters.  Each



was concentrated and extracted with nitromethane (NM); the MM extracts



(fraction B__) were evaporated to dryness, redissolved in n-hexane



and analyzed by GC.  All  PAH through the dibenzanthracenes (MW 278) were




found to be in the initial 2.5 liters fraction.  Since fluorescence was.



observed in the 2.5 to 3.5 liter fraction, including the last 100 ml of



this fraction, higher molecular weight strongly fluorescing PAH are no



doubt responsible for fluorescence at high elution volumes.  Sample 19^72



contained the highest amounts of PAH.  Thus, it was acceptable to term-



inate elution  after 2 liters of solvent on all other samples (see Table IV).
                                23

-------
                             TABLE IV

       VOLUME OF 8:1 HEXANE/BENZENE USED TO ELUTE FRACTION B



Sample Number                           Volume 8;1 Hexane/Benzene,  ml

     19U66                                        I600a

     19^67                                        2000

     19U69                               .         2200

  .   19U70                                        2100

     19^71                                        2000

     19^72                                        3500

     19^7U                                        2050

     19^76                                        1250a
ft
  No fluorescence observed when these volumes had eluted.

-------
     2.  Gas Chronatographic Analysis



         Excellent separations  of PAH  were achieved employing Dexsil 2%



and 6% columns.   Figure 1 shows a gas  chromatographic  trace representing



a typical analysis of a ten component  PAH standard mixture.  Table V



gives the structures and properties of these compounds.   Figure 2 shows



a gas chromatographic trace representing the analysis  of sample 19^72;



the sample represents the first 2.5 liters of 8:1 hexane/benzene from



the LC column (see page 9).




     In Figure 1 it can be seen that phenanthrene and  anthracene (peaks 1



and 2) are partially separated, benz[a]pyrene (BaP.) and benz[e]pyrene



(BeP) (Peaks 6 and 7"1 are not separated and dibenz[a,h]- and dibenz-[a,c]-




anthracene (peaks 9 and 10) are not- separated.  These  observations repro-



duce the data of Lao et al.



     Components  in the standard mixture were chosen to represent a PAH



envelope that might be observed in sediment samples.  Phenanthrene and



anthracene are three ring PAH with molecular weights (MW) of 178.  The



dibenzanthracenes are five ring PAH with MW's of 278.   In between there



can be PAH with varying number of rings, ring structures, and substitu-



tions on the rings.  Over 100 PAH are  known between MW 178 and 278. •



Effective separations are both critical and difficult; with caution they




can be accomplished  and compound assignments be made.



      Figure 2 shows  assignment of PAH to components of the standard mix-



ture.  In addition to comparing gas chrbmatograms  of  standards  and samples,



a comparison with the data of  Lao  was useful.in  chromatogram  interpretation.
                                25

-------
                                               26

FIGURE I.  Recorder trace of detector response in gas chromatographlc analysis of PAH standard
           mixture using 6$ Dexsil 300 column; Attenuation and amplifier Bettings are given in
           the figure as IB recorder chart speed; Peak numbers correspond to the following PAJ1:
           1. phenanthrene; 2. anthracene; 3. fluoranthene; *4. pyrene; 5. benzfa]anthracene;
           6 and 7. benz[a]pyrene -and benz[e]pyrene; 8. perylene; 9 and 10. dibenz[a,c]anthracene
           and dibenz[a,h]anthracene.

-------
FIGURE II.  GC trace for analysis  of sample 19!472,  0-2.5 liter, 8:1 hexatie/benzene fraction.
            Attenuation, amplifier,  and  recorder settings are as indicated in the figure.
                                                                                   * T » 1 * .«.«.• «.« 1 * • * » •

-------
                                                   28
                                              TABLE  V
                STRUCTURES AND PROPERTIES OF CERTAIN POLYNUCLEAR AROMATIC HYDROCARBONS
          PAH
           Molecular
Formula      Weight
                                                            Structure
                                                           Carcinogeni c
                                                             Behavior
Phenanthrene



Anthracene



Fluoranthene



Pyrene



3enz[a]anthracene



Eenz[a]pyrene



Benz[e]pyrene



Perylene
                          Vw       178
                          ci4Hio
                          C16H10
                          C16H10
                          C18H12
                          C20H12
                         C20H12
                         C20H12
Dibenz[a,c]anthracene     CppHlU
Dibenz[a,h]anthracene
178



202



202



228



252



252



252



278



278
                                 oo

                                   O
                                                                           o
                                                                            ++++

-------
a Ratings of carcinogenic cehavior obtained from reference 20';  (-)
  indicates not carcinogenic; ( + ) uncertain or weakly carcinogenic,
  ( + )  carcinogenic; (-+,  +++, +-H-+) strongly carcinogenic.
                                 29

-------
     The first -.r.a.jor pe?-k (labeled 1 and 2) in Figure 2 represents phenan-



threne and anthracene.  The assignment of these compounds is clear.  However,



since only one 1C pea.-: appeared it is probable that either phenanthrene or •



anthracene predc~ir.2.-es.   One cannot distinguish unequivocally between the two



and quantitation IT.UST; be  nade as the sum of phenanthrene and anthracene;' thi6



situation obtains for all other samples.  For sample 19^-72 GC/MS experiments



suggest that the r.a.jcr peak (l and 2) is predominantly phenanthrene with a



minor contribution from anthracene (see Figure 5, page k2).



     The peaks labeled 3 and h can be assigned to fluoranthene and pyrene.



Further support for these (and other) assignments -will be given in the



GC/MS data to be presented below.  Peak 5 certainly represents at least



two overlapping peaks.  This is consistent with the observation of Lao



et al  which notes that benz[a]anthracene is not separated from chrysene



and triphenylene using the Dexsil GC system.  Peak 5, then, should be



assigned to these three compounds; no other PAH of MW 228 elutes near these com



pounds  and GC/MS data show that the peak is due to PAH having molecular


                                                               po
weights of 228.  Benz[a]anthracene and chrysene are carcinogens   while



triphenylene is not.



     Peaks labeled 6 and 7 are assigned to EaP and EeP; peak 8 is assigned



to perylene.  GC/MS data show these peaks to have molecular weights of



252.  The doublet consisting of a large and a small peak elating before



BaP and BeP also have molecular weights of 252 (see below).  Based on the



data of Lao  the compounds represented by these peaks may be the benzofluor-



anthene isomers.
                                30

-------
     Peaks 9 ?-r.d -"'' ^"e assigned, to the dibenzatythracenes.  Other PAH

                                          g
vhich elute i:i "he ~.iben~an'hracene region  and vrhic'n may be present in


certain sairrples are ^he cenzocnrysenes, picene, the benzoperylen.es,


o_-phenylenepyrer.e , ar.d the bensotetraphenes .


     Table VI reports the concentrations of'FAH determined in sediment


samples (from frac'icr.s ET_ ; .  Matching of individual components in -the


samples was not always as sinple as in 19^72 described above and in Fig-


ure 2.  For samples vhere certain components could not be compared  with the


standard mixture a value is not reported in Table VI.


     Samples 1QU66 and 19^76 display the lowest levels of PAH.  This is


consistent with their Low oil and grease values and the  fact that fluor-


escence terminated after 1-1.5 liters of solvent in collecting  fraction B.


It is also consistent with  the locations of these two sampling  sites.


Sample 19^-72 being just downstream from outfall 002 shows by far the high-


est concentrations of PAH.  No explanation can be offered for the absence


of lower molecular weight PAK in sample 19^71-


     Sample 19U71* was different than all other sediments collected.  Frac-


tion A showed an apparently high paraffinic content.  This was  observed by


analysis of fraction A employing the Dexsil column.  Fractions  A of several


samples were scanned for qualitative information as to paraffinic content.


Sample 19U?1*- contained at least a ten fold higher paraffinic level  than any


other sample examined.  In addition examination of Fraction ETT for PAH


showed a different shaped GC elution pattern.  The GC trace in  Figure 2

-------
                                                    TABLE  VI

                           CONCENTRATIONS OF PAH DETERMINED IN LORAIN SEDIMENT SAMPLES


                                                Concentration, mg/kg (dry sample weight)a
Sample
Number
19*i66
19*i67
19*l69
19)i70
191*71
19)172
19!i7*l
19)176
Phenanthrene
& Anthracene
0.^3
2.8
2.6
3-2
b
18
~3b
0.68
Fluoranthene
b
2.5
3-9
3.3
e
29
b
0.75
Pyrene
b
2.9
5.2
h.k
e
30
b
0.61
Benz[ a] anthracene
0.83d
i.6d
2.9d
2.8d
3-id
2*1
b
1.2d
Benz[a]pyrene &
Benz[elpyrene
0.91
1.5
It. 6
2-9
li.Y
.19
1 . 1
1.5
Perylene
0.10
o.oH
e
e
3-9
2.0
e
0.35
Dibenzanth race nes
0.67C
0.37
2.6
e
3-9
I?.
c
o.66c
Q
  Data calculated by comparison of PAH in the sample fraction B T  and a standard mixture  of ten PAH.



  Overlapping peaks in the gas chromatogram precluded analysis of  this PAH.



  Other major peaks occurred in the dibenzanthracene area of the GC.   Concentration  reported  represents only

  GC peak at retention time of dibenzanthracenes in the standard mixture.



  The GC peak clearly represents more than one compound, as reported  in reference 6.   The  values  given  are  for
  the total PAH present based on the benz[a]anthracene standard.


g
  Less than detectable.

-------
for 19^72 is typical cf samples analyzed except 19^7^.  Consistent with



our observations concerning sample 19V/U is the fact that U.S. Steel



apparently discharges waste oils from outfall 001; sample I^kjk was taken



0.2 miles downstream from this outfall.



     3.  Gas Chromatcgraphic/Mass Spectrometric (GC/MS) Analysis



         Sample 19^72 was examined in detail by GC/MS.  In addition the ten



component FAH solution was analyzed.  Both analyses employed the 3$ Dexsil



column.  The "real--cine" and reconstructed gas chromatogram (RGC) for



the standard mixture showed, separations to be identical to these shown



in Figure 1.  Mass spectra for the standard compounds were consistent


                           6 21 22
with spectra for known PAH. '   '



     Figure 3 shows the "real-time" gas chromatograra for the analysis



of sample 19^72.  (All further mass spectral data reported are for this



sampled.  The GC is essentially the same as the trace shown in Figure 2.



In GC/MS analysis the total ion current is plotted, as in Figure 3, after



each spectrum scan during the  run.  Over sixty peaks can be counted.



Figure U shows the reconstructed gas chromatogram (RGC) of the same



run shown in Figure 3-   The data system has normalized the largest peak



in the RGC to an amplitude of  100.  Because of the way the data system



acquires and processes  information relative peak heights between the



"real-time"  and reconstructed  chromatograms are not always the same.



Further explanation cannot be  given in this section,  owing to  space limi-



tations.
                               33

-------
FIGURE 3:   "Real-tine" gas chroraatograa of GC/MS analysis of sample 19^72,




           Times in minutes and seconds written above each peak represent




           retention tines of the eluting components.






FIGURE U:   Reconstructed gas chroinatogram for GC/MS analysis of sample




           19^72.






FIGURE 5:   Reconstructed gas chromatogram for .GC/MS analysis of sample




           19^72 employing a limited mass search for ions m/e 178, 202,




           228, 252, and 278.






FIGURE 6:   Reconstructed gas chromatogram for GC/MS analysis of sample




           19^72 employing a limited mass search for 'ions m/e 192, 216,




           2U2, 266, and 292.

-------
 35




FIGURE  3
                                   J. CJ Id W{3 3
                                                                      I!

-------
 36
FIGURE 3 (COOT)
                                                   .40-14*103

-------
 37






FIGURE 3 (CONT)

-------
                                                                              38

                                                                           FIGURE
        C-131TZ 3>CE>OL IfSrOS
  8
  B_
  S.
  P.
  8_

£
  R.
  8.
       !„..,... ,,.,,,,.MI..,,,.,,,	p.,.,....,...,!...,,,...,.., ...,,..,	,
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-------
                                                           39




                                                          FIGURE U (CONT)
310
       '
                                                                                     « sa          a   i

-------
                           Uo



                          FIGURE  U (CONT)
•T-i'-l	l""i""l	'"I	I	'"I	|'"i'"|

-------
     Figure 3 gives ?.r. RGC limited mass search fcr the parent ions cor-




responding to the cc-pcunds in the PAH standard mixture.  The ions




causing the RGC-peaks are indicated above each individual peak.  A peak




corresponding to n/e 278 is not observed since it is small compared to




the 202 peak on which the RGC is normalized.  The mass spectra of PAH




ajjnost always display intense parent ions.  The parent is usually the




base peak.  Thus performing a limited mass search for the parent is an




effective way of defining particular PAH.  A cluster is often present




about the parent representing the P-l, 'P-2, P-3, P+l, and P+2 masses; - that




is the parent less  one,  two,  and three and plus one and two arr.u.




     Figure 6 gives an RGC limited mass search for the methyl-substituted




derivatives of compounds in the PAH standard mixture."  Thus the unsubsti-




tuted parents -t-lU amu were searched.  Again, the molecular ions causing the




RGC peaks have been indicated.




     A limited mass search for dimethyl derivatives, the unsubstituted




parents +2$ amUj was r.ade but  the data were not conclusive and are not being




presented.




     Searching for unsubstituted ions in the standard and their-methyl




derivatives produced thirty-three separate GC/MS peaks indicating at least




that many compounds.  In certain cases two or more PAH were present for




a specific HGC peak indicating more than one PAK was eluting from the




Dexsil column into the ion source at the same time; this observation




is quite reasonable.  Searching for the dimethyl derivatives produced an




additional eight distinct GC/MS peaks.
                                Ul

-------
                                                                           1*2
                                                                          FIGURE 5
       t-12-rrz 3>CEX1U KB-SG
 8.
 S_
 P
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                                                                                                                        ZED  zco 270  CDS  zsi 200  310
                                                                                                                        .AH-iu HMO a

-------
                                                                         FIGURE  5 (CONT)
lea
• llllillll I	I i I  I   I'" I'  I'  'I 	 I  I "'I	I 'I' I   '  I
 jC3  3Q3  1C3  118  tZD  130  Vt9  1S3 1CO T78  K
S  SIO  SZ8  ba £«  SSO  ECO  £78  SSO  EO 633 «13  6Z3

-------
                                                             uu
                                                            FIGURE 5  (CONT)
i i  |  •  i
i|i if• ji
 era  «23 ccs  era  'twa csa 7ca  TIB  •KD '"no  713  7S9 7C9  770  780
                    I
I""!—-T I	       «  |   •  |  •  |  *  I  '   »
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                                                                                                            310 2O5

-------
 8.

9s.
                                                          FIGURE 6
 B
 8.
      LIMITED MASS
        H/C  iqa.au,
'-r-r.
 0
so'  to
              M/QQI
w'  aa'  in no iza  li "Aw ioi ici ni  iw  isi
                                                                                  210
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-------
                                                              U6




                                                            FIGURE 6(CONT)
It.
««
                                                                   '«" TO""'-!^'
                •*fflM«M»M  •_„..»».•  tWM-M*nM
                                                | MMMIflM
                                                                                          OT bo  s
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-------
     Mass spectral iata shew that the sample extract contained almost exclu-




sively ?AK.  The largest peal-: in the RGC (Figure k) was not a PAH but a




phthalate derivative with a strong base peak of m/e lU9.  The presence of




this compound cannon ~e explained.




     Presentation of all niass spectra is precluded by space limitations.




In Figure 5, however, it can be seen that peaks for phenanthrene +




anthracene (m/e 173), flucranthene (first m/e 202), pyrene (second large




m/e 202), benz[a]anthracene and probably chrysene and tetraphene (large




broad m/e 228), benz[a]- and benz[e]pyrene (large peak of second m/e 252




doublet1) and perylene (last m/e 252 peak) are present.  Comparison should




be made with Figure 2.




     Presentation of selected spectra is useful.  Figures 7 through 11




show the mass spectra of major RGC peaks (Figure 5) having m/e 252 as




the base peak - the benzfluoranthene, benzpyrene, perylene area.




Figures 12 through 15 show the mass spectra of major RGC peaks (Figure 6)




having m/e 2U2 as the base peak.  These likely represent methyl-PAH which




are not identified.  Other unsubstituted PAH having molecular weights of




2^2 are possible but unlikely.  It should be noted the computer was




allowed to subtract background from spectra of interest.  Thus in Figure 7




spectrum 526-520 signifies that spectrum 520 has been subtracted from 526.




     Another method in identifying and confirming molecular formulas by




mass spectrescopy employs isotope abundances of the common elements.  For




PAH one need consider only carbon and hydrogen.  For every carbon atom,

-------
FIGURES 7,  8,  9,  10,  11:   Mass  spectra of PAH having n/e  252  as  the base




                          peak  in sample  19^72.








FIGURES 12, 13,  1^,  15:    Mass  spectra of FAH having m/e  2U2  as  the base




                          peak  in sample  19^72.

-------
                       sc«  -   sj
                                                                              FIGURE  7
  B
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0.
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                                                                                                    2ZO  Z30
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-------
                                                                    50

                                                                  FIGURE 8
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                                                                             51

                                                                          FIGURE" 9
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-------
                                                                            52


                                                                        FIGURE 10
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-------
                                                                        53

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                                                                    56

                                                                  FIGURE  lU
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     E-15TTZ >CEX1L 1GS-23S
                            JJH  JrJ.itl	t
SO  CO   TO   SO
in no  >a>  «, ',« ',»  in no  ira  i» zw ttB'ea za T«  CT
,' i..
cm cw
                                                                                                                           a.a
                                                                                                                                    aad'

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                                                                           57




                                                                         FIGURE 15
     TITIT1





     T.'..VL  10" C''K
                     r»i*4t> sr.nKxiL I»S-«»S»«/N MOLD  in<-i>*/v  tnoiw
  8
  3.
|SJ


hSL
|SJ
       STCTPUI oretn icz  -



       C-1M7Z XXX1L 1CS-235
                                                                        b
                                                                                                            n  i"
                                                                                                              W

•: . -.
                                                                ls8
*" ***  M  "*
                                              fc8 3CO  «10  3M 3»

                                                                                     	..l.

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                                                      13
ca 1.08$ of the nolecules contair.ir.fi carbon possess a   C atom and produce a




P + 1 (parent -1) pesJc. Za.ch atom in an ion contributes an amount to the




intensity of the isotope peak that is equal to the relative abundance


                            2k
of the isotope of that atom.   "Thus if a strong molecular ion is present,




as in the case of PAH, the P+l abundance is useful in identifying a par-


                          ?\±

ticuiar molecular formula.    The P+2 is also considered, for there exists.




a probability that two  JC atoms exist in a single ion.  Values of P+l




and P+2 have been calculated for the various -.olecular formulas and are



                    23 25
presented in tables.




     Table VII list ?+l and P+2 isotope abundances for selected PAH of




interest.  The values are theoretical except for two molecular formulas




which represent experimentally determined literature data.  Table VIII gives the




P+l and P+2 values for the PAH in the ten component .standard mixture.




Excellent agreement is observed.




     Table IX gives the values found for certain major PAH present in




sample 19^72.  The values were obtained from spectra defined by the




reconstructed gas chromatograms shown in Figure 5 for unsubstituted and




Figure 6 for methyl substituted PAH.  Agreement for the unsubstituted PAH




is good.  In a sample containing often overlapping GC peaks one would not




expect agreement to be ideal.  This is especially true for the P+2 isotope



abundances which is a second-order effect.  In addition,  spectra were defined




by limited mass reconstructed gas chromatographic searches.  Overlapping




compounds present which are not seen in the limited mass search can easily

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interfere with isotope abundar.ee calculations.  The total amounts of




methyl substituted PAH present are considerably smaller than unsubsti-




tuted polynuclears.   Thus, one would not expect isotope abundance calcu-




lations to be nearly as satisfactory for these cases; this is exactly




the situation.
                                 59

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                             TABLE VII
             ISOTOPE A2UIvDANCES FOR MOLECULAR  FORMULAS
             CORKESFO;TDING TO PAH OF
•Formula
C1UH10
C15H12
cl6Hio
C17H12
C18H12
C19H1U
C20H12
C22"lU
Molecular
Weisht
178
192
202
216
228
2U2
252
278
P+l
15.29
16. Uo
17.^5
18.56
19.6*1
20.76
21.9
23.5
P+2
1.09
1.26
1.U3
1.62
1.82
2.0^
2.U
3.2
9.
  Values are theoretical (reference 23) except  for  MW 252 and 278;
  these values are experimental findings  (reference
                                  60

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                            TABLE VIII

            ISOTCFE Ar.lJT-rDA.NCE FOR PAH STANDARD MIXTURE
Compound Assigned
Phenanthrene )
Anthracene )
Fluoranthene
Pyrene
Eenz [a] anthracene
Benz[a]pyrene )
Benz[e]pyrene )
Dibenz [a, c] anthracene )
Dibenz[a,h]anthracene )
Formula
C1UH10
ClUH10
C16H10
C16H10
C18H12
C20H12
C22Hli|
Molecular
Weight
178
178
202
202
228
252
278
P+l
15. 2a
15. 5a
17.6
17.7
19-7
21.1b
23. 5b
P+2
l.3a
l.Ua
1.6
!•?
2;i
2.2b
3.1b
Q
  Phenanthrene and anthracene were slightly separated  as  in  Figure  U.
  Values for each side of the peak were calculated.


  Values given are averages for several sections of each  peak;  each
  peak represents two compounds J  see Figure 2.
                                 61

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                          TABLE IX

 ISOTOPE A?.UT!DA::CIS  rOR  CERTAIN PAH FOUND IN SAMPLE

                              PAH
Spectrum                 Parent Peak           P+l            P+2
A. Unsubstituteci
211-206
215-206
337-333
3**3-333
353-3^9
1*33-1*30
1*36-1*30
526-520
51*0-520
563-557
566-557
582-576
78U-775
821-805
B. Methyl Substituted
268-262
278-262
372-369
375-369
378-369
386-383
393-390
1*6-1*1*2
1*57-1*53
1*60-1*53
1*62-1*53

178
178
202
202
202
228
228
252
252
252
252
252
278
278

192
b
216
216
216
216.
216 ,'
2U2
2l*2
2l*2
2U2

15. U
15.6
18.3
16.8
17.9
20.1
20.0
20.3
21.9
19.6
20. U
21.5
26.7
19-3

18.2
b
16.0
16.6
16.0
b
b
19-6
20.7
22.1*
23.2

1.1
1.2.
1.1*
0.9
2.0
l.U
1.2
2:r
1.9
1.9
1.9
2.0
e
e

1.2
b
c
U.5
c
b
b
c
c
c
<~c
                                62

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                          TABLE  IX  (CONT)


a.  Refer to Fig-i^re ?  fcr unsubstituted  FAH and Figure 6 for methyl
    substituted "A:-!.

b.  Overlapping of tv;o or :nore ccmpour.ds prevented analysis by this method,

c.  P+2 value sns.ll cr net nresent.
                                  63

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                            DISCUSSIC::



A.  SURVEY FI:.^::;^



     Water sa.-r.ples vere :'cu::d not to be contaminated to any significant



degree.  The cc>e rev.-er ''CC2^ did show small quantities of organic com-



pounds and likely ccr.«ai::ed lover molecular weight polynuclear aromatic



hydrocarbons 'PAH;.



     The data of Table I shows the presence of substantial amounts of



hexane extractable organic residues in sediment samples.  All Lorain



sediments vrere surface samples.  ji-Hexar.e was vised as the extracting



solvent for two reasons.  First it was felt that use of a halogenated



solvent such as Freon would later interfere with gas chroinatographic/



mass spectrcmetric analysis of the residues for PAH.  In addition,



n-hexane was the extracting solvent used in the U. S. Steel Calumet survey



and it was felt that comparisons between the two surveys would need to be



drawn.


                 26
     Calumet data   show "oil and grease" values  as high  as'120,000 ppm



for one sample and over 60,000 ppm for many others; some of these were



core samples.  Lorain data show, overall, slightly lower oil and grease



values.  This could be consistent with the fact that the Black River has



recently been dredged.



     Reasonably, the highest "oil and grease" values in the Lorain survey



were found in the vicinity of the plant discharge points.  The lowest



values determined were at the harbor mouth and 2-3 miles downstrean from



the outfalls.  The lev; value for sample 19U68 cannot be explained.

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     It is terr.p^ir1.:; zo try to drav: analogies between amounts of organic



residue and particular classes of organic compounds.present in sediment



samples.  In certain cases this can no doubt be done; however, caution



must be exercised.  ~or example while samples 19^72 and 19^?U yielded



similar amounts of organic residue, their chemical compositions are



quite different.



     Table VI and ether data presented in the Results section shows



that a broad spectrum, of FAK is  present in the 'sediment samples.  The



highest concentrations are in the vicinity of the plant outfalls with



the levels in 1QV72 being 2-10 fold greater, depending on specific PAH,



than for any other sample analyzed.  Lowest values are found for sample



19U66 (harbor  mouth), 1QU67 (center of the river mouth), and 19^76



(0.2 miles upstream from outfall 001).  It is reasonable that PAH were



found in all samples since the sampling sites were in close proximity



to the plant.  A lake effect is known to occur carrying water upstream


                                         27
at least five miles from the river mouth.     Because this survey rep-



resented the first attempt in this laboratory to quantitatively analyze



for many PAH, one probably should place wide tolerance limits on the



accuracy of the data in Table VI.  The data certainly reflect  the levels



of PAH in the analyzed, samples; however, errors of + 15$ would not be



unexpected.
                                65

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     Many of tr.e ~AH round in the sediment samples are without question

carcinogenic.  T'r.ese include not only compounds reported in Table VI

(see also Table "•/'• .   Since ?. broad envelope of PAH was  shown' qualita-

tively to be present, rr.ar.y unidentified compounds  containing  four or

more rings, are r.o doubt carcinogenic.  In particular are the methyl sub-

stituted. FAK which have been shown to possess a high degree of carcino-

genicity.5'20


3.  OTHER COMMENTS

     The general philosophy taken at the outset of the survey was to

analyze for a broad envelope of PAH rather than for one or two compounds.

This approach proved to be successful and we believe that future surveys

should be handled in a similar fashion.


     The analysis for complex mixtures of PAH constitutes one of the most •

difficult determinations in organic chemistry.; the data presented in this

report are clear evidence of this fact.  Optimum conditions for separating

PAH from other classes of organic compounds had to be determined.  GC and

GC/MS analyses techniques had to be perfected.  The PAH fraction (B,.,) is

complex and requires careful and detailed analysis and interpretation.

These criteria were met and meaningful data were obtained.

     In subsequent'surveys, in other geographical  locations,  our capa-

bilities will be extended.  For example, we intend on obtaining a stock of
                                     /
PAH primary standards available from'the Canadian Air Pollution Control'
                                                   r,0
Directorate, Department of the Environment, Ottawa.    These will allow

us to quantitate upwards of 50 PAH.
                                66

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                              REFERENCES

  1.  P.  R.  Clifford,  ''Organic Analysis of the Grand Calumet Oil and
      Grease Samples.'1 I-fr'IC-Cincinnati Report, January,  1973.

  2.  "Standard Methods for the Examination of Water and Wastevater,"
      Thirteenth Edition, I-'.J. Taras ,  A.  E. Greenberg, R. D. Hook, and
      M.  C.  Rand,  Eds., American Public Health Association, Washington,
      D.  C., 1971, pp. U09-U13.

  3.  D.  Hoffmann, ar.d  E. L. wynder,  Cancer, 27, 8U8 (1971).

  U.  D.  Hoffmann and  E. L. Wynder,  Anal. Chem. , 32, 295- (1960^ .

  5-  £.  Wynder and D. Hoffmann, "Tobacco and Tobacco Smoke," Academic
      Press, New York, 1°67 pp. 330-3UU.

  6.  R.  C.  Lao, P.. S. Thomas, H. Oja, and L. Dubois, Anal. Chem., 1+5,
      903 (1973).

  7.  J.  W.  Eichelberger, L. E. Harris,  and W. L. Budde, ibid , U6, 227
      (197*0.

  8.  W.  L.  Budde et al ibid. , in press

  9.  "System 150 GC/MS Data Processor:   System Specifications," System
      Industries,  Inc., Sunnyvale, California,
 10.  J. B. Knight,  "Computerized GC/MS For Quick Qualitative Identifi-
      cation of Hydrocarbon 'Types Included in Chromatographic Peaks,"
      Finnigan Corporation Application Tip No. U3, Finnigan Corporation,
      Sunnyvale, California, 1972.

11.   K. Bhatia, Anal.  Chem., U3, 609 (1971).

12.   W. Lijinsky,  I.I. Dorasky, and J. Ward, J. Gas Chrom., 3, 152 (1965).

13.   V. Cantuti, 0.- P. Cartoni, A. A. Leherti, and A. G. Tori, J.
      Chromatography, 17, 60 (1965' .

lU.   N. Carugno and S. Rossi, J. Gas/Chroin. , 5_, 103 (1967).

15.   H. J. Dawson,  Anal. Chem., 9, 1852 (196U) .

16.   H. J. Davis,  Talanta, 16, 621 (1969).
                                 67

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17.  K.'j. Davis,  Anal.  Chem.,  Uo,  15S3  (1968).

10.  D. A. Lane,. H.  K.  Mre,  and M.  Katz,  it id.,  U5,  1776  (1973)

19.  R. F. Skinner,  J.  B.  Knight, D.  M.  Taylor,  and  E.  J.  Eonelli,  "The
     Determination of Folycylic Aromatic Hydrocarbons in  Air Pollution
     Samples," l-'ir.nigan Corporation Application  Tip  No. 52,  Finnigan
     Corporation,  Sunr.yvale, California.,  1973-

20.  "Particulare Folycyclic Organic  Matter,"  National Academy of Sci-
     ences, Washington,  D. C.,  (1972),  Chapter 2.

21.  L. Dubois, R. C. Lao, and  R. S.  Thomas,  "Mass,  Ultraviolet,  and
     Fluorescence Spectrometric Data  of Polycyclic Hydrocarbons Found
     in Urban Air and Cigarette Smoke,"  l6th  Spectroscopy Symposium,
     Montreal, Canada,  1969.

22.  "Atlas of Mass Spectral Data," E.  Stenhagen,  S. Abrahamsson, and
     F. W. McLafferty Eds.,  Wiley-Interscience,  New  York,  1969.

23.  R. M. Silverstein and G. C. Bassler, "Spectrometric  Identification
     of Organic Compounds,"  Second  Edition, John Wiley, New York, 1967,
     Chapter 1.

2U.  S. R. Shrader,  "Introductory Mass  Spectrometry," Allyn and Bacon,
     Inc., Boston, 1971, Chapter.1.

25.  J. H. Beynon and A. E.  Williams, "Mass and  Abundance Tables  for Use
     in Mass Spectrometry,"  Elsevier, Amsterdam, 1963.

26.  "Grand Calumet River Sediment  Measurements  -  Collection and  Analysis,"
     NFIC-Cincinnati Report, 1972.

27.  T. Braidech, private communication.

28.  J. L. Monkman,  "Annual Report, International  Bank of Polycyclic
     Aromatic Hydrocarbons," World  Health Organization, Lyon, France,
     December, 1973.
                                68

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