CHEMISTRY LABORATORY MANUAL
        BOTTOM SEDIMENTS

          COMPILED BY

      GREAT LAKES REGION
COMMITTEE ON ANALYTICAL METHODS

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                                      TABLE OF CONTENTS
                                                                                Page
             FOREWORD                                                            iii

             ACKNOWLEDGEMENTS                                                    iv

             INTRODUCTION                                                        1

             CHEMICAL OXYGEN DEMAND                                              5

             CYANIDE                                                             7

             IRON
                       Total and Soluble Ferrous" Iron                           12
                       Total Ferrous  Iron by Titration                           16

             METALS
                       Atomic Absorption                                        18
                       Polarographically                                        21

             NITROGEN
                       Ammonia                                                  28
                       Nitrate and Nitrite (AutoAnalyzer)                        32
                       Nitrate (Manual)                                          36
                       Organic and Total Kjeldahl                               38

             OIL AND GREASE                                                     k2

             OXYGEN DEMAND AND CHLORINE  DEMAND
                       Immediate Dissolved Oxygen Demand and                    kk
                            Biochemical  Oxygen Demand
                       Oxygen Uptake                                             hj
                       Chlorine Demand                                          53
             •PESTICIDES
                      Preparation of Sample                                    56
                      Gas Chromatography with Electron Capture                 59
                      IMicrocoulometric Gas Chromatograph                       64
                      Thin Layer Chromatography                                69
                      Infrared                                                 75

I           PHENOL                                                             77
            PHOSPHORUS
                      Total Soluble                                            79
                      Total                                                    82

            SOLIDS - Total (%) and Volatile ($)                                85

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                              TABLE OF CONTENTS








SULFIDE




SILICA




MANGANESE




OXIDATION-REDUCTION POTENTIAL MEASUREMENTS


Page
87
92
98
101













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                                  FOREWORD


     Water pollution control agencies during the past two or three years
 have become concerned over the U. S. Army Corps of Engineers dumping dredg-
 ings from shipping channels and harbors into the deep waters of the Great
 Lakes.  The state and federal agencies and the Corps of Engineers recognized
 the necessity of knowing the chemical content of these dredgings or bottom
 sediments.


     The Corps of Engineers then requested that the Federal Water Pollution
 Control Administration sample and analyze all bottom sediments in areas to
 be dredged, prior to dredging.  As a result of this request, the Lake Michi-
 gan Basin Office developed analytical methods suitable for the analysis of
 these bottom sediments.  It was not long before other FVPCA Basin Offices
 in the Great Lakes Region requested the Lake Michigan Basin Office's methods.
 Other Pasin Offices soon modified the Lake Michigan Basin Office's methods
 to suit their particular need.


     In October 196? chemists of several agencies in the Great Lakes Region
 were requested to develop uniform sampling and analytical methods.  A meeting
 including FVPCA Pasin Offices, U. S. Army Corps of Engineers,  International
 Joint Commission and U.S. Lake Survey was held in Cleveland to discuss
 sampling and analvtical methods and establish uniform methods.  A committee
 was appointed to compile a chemistry manual for bottom sediments.  The
 committee consisted of members from all of the above agencies.


     After extensive correspondence, exchange of methods among the committee
 members and several committee meetings,  the present manual was compiled.


     This manual is being used by all of those agencies  mentioned above in
 the Great Lakes Region and many other federal,  state, municipal,  consulting
firm and university laboratories.
                                            Frederic D.  Puller
                                            Chairman and Editor
                                            Lake Michigan Basin Office
                                    111

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                           ACKNOWLEDGMENT
In addition to those chemists mentioned above, credit is given to all
chemists in the region for their contributions of thoughts and suggestions.
                                     iv
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Various tests as given in Standard Methods for the Examination of Water
and Wastewater have been modified to better suit Great Lakes Region's
objectives and needs.  In some instances, new methods have been devised
as superior to those in Standard Methods.                                       •

For these endeavors by members of the many laboratories in the Great
Lakes Region, full credit is given.
                                                                                I
Original Methods

     Total Ferrous Iron, by C. Ross and C. Potos,
     Lake Erie Basin Office

     Pesticides, by W. D. Johnson and G. Frye                                    m
     Lake Michigan Basin Office                                                  •

     Chromium by Polarography, by H. S. Whitworth and E. T. King                 —
     Lake Michigan Basin Office                                                  •

Modified Methods

     Toxic Metals, including preparation of sample and methods                   |
     for Cu, Cd, Ni, Zn, Pb, Cr, Al, and Mn, by E. T. King,
     R. S. Whitworth and S. Zarbin                                               _
     Lake Michigan Basin Office                                                  •

     Cyanide, by E. T. King, Lake Michigan Basin Office

     Sulfide, by E. T. King and S. Zarbin, Lake Michigan Basin Office            •

     Total Phosphorus and Total Soluble Phosphorus, by F. D. Fuller,             •
     R. H. Hall, and M. Huston, Lake Michigan Basin Office                       J

     Silica and Manganese, by A. C. Smith, Lake Erie Basin Office                _

     Total Iron, by R. S. Whitworth, Lake Michigan Basin Office                  ™
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                            INTRODUCTION TO
                         BOTTOM SEDIMENT ANALYSIS
1.   General Discussion

     Bottom sediment analysis gives information as to the chemical and
     biochemical characteristics of the overlying water mass and the
     immediate underlying sediment itself.

     The productivity of a stream or lake is reflected in the chemical
     composition of the bottom sediments.

     Comprehensive bottom studies shov potential pollutional loadings
     of toxic materials, nutrients, and organic substance.

2.   Sampling

     2.1  Bottom sediment samples are collected with a Petersen or
          Shipek dredge lowered slowly in order to cause the least possible
          disturbance to the surface sediment (water-sediment interface).
          A satisfactory core sampler would be more desirable.

     2.2  When hopper dredging is employed, inflow grab samples are taken
          from two forward and two aft  inflow chutes over an eight hour
          period and composited.  The composite is thoroughly mixed and
          samples withdrawn.  Overflow samples are collected at four over-
          flow weir locations and composited in the same manner.  Sampling
          from the hopper itself is accomplished by allowing a 20-quart
          bucket to remain in the bottom of the hopper at four locations
          for five minutes and compositing these grabs similar to the
          inflow and outflow samples.

     2.3  When bucket dredging is used, five samples from each compartment
          would be taken after the barge is filled.  A one-inch diameter
          aluminum tube, ten feet long will be used to take samples.  One
          composite is made for each barge and two barges per week.

     2.k  When hydraulic dredging is employed, grab samples are taken from
          the discharge pipe and composited.  Bie sample is taken from the
          well-mixed composite.

3.   Preservation and Storage

     All samples should be well iced and no preservative added.  If the
     samples can not be analyzed the day they are collected,  they should
     be frozen as soon as possible.  Sample containers should be filled
     only three-fourths full to prevent breaking, when frozen.

k.   Preliminary Preparation of Bottom Sediment Samples

     The sample is passed through a 2 mm (approximately 10 mesh) sieve
     by rubbing with a rubber stopper, if necessary.  This removes large
     pieces of foreign material such as stones and twigs.

     The sieved sample is blended to a homogeneous mixture in a
     blender for 2 minutes at high speed.  The sample is now ready for
     analysis.

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SCHEMATIC ANALYSIS OF SEDIMENT
E^, and pH performed in field
SIEVED BLENDED NO ADD SAMPLE
PHENOL •£ 10~50g I 1 10 p METALS
~vr» — .3~lg «5~lg /1/VTV
— 5-10ff .S-.8e > „ =
O T f^ rr G» T f\f9 li
_^..S-l2 lOff
-N p* * "^ 	 	 	 	 > CYANIDE
1 10 g
Residue Dry in oven at 103°-105°C - overnight
digested
Org.-N Percent: Solids

ignite residue at 600°C for one hour
I
Volatile Solids

Decomposition and dehydration of residue using HC1-HNO
i
Silica

Filtrate
Iron Manganese

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FIGURE 1 M

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Introduction to Bottom Sediment Analysis

5«   Schematic Analysis

     If the sample has been frozen, thaw the sample and proceed with
     preparation of sample.  Several portions of the wet, sieved,
     blended sample are weighed depending on the parameter concentration
     expected (Figure l). The amount of sample necessary for any given
     test is determined by the analyst based on test sensitivity and
     concentration in the sample examined.

     5-1  One weighed portion of wet sample is placed on aluminum
          strips for the BOD test.

     5.2  A portion of the sample is weighed for the COD test.  Fifty ml
          of distilled water is added, followed by three drops of con-
          centrated sulfuric acid.  The acid preserves the sample until
          the analysis can be completed.

     5-3  A portion of the wet sample is weighed for nitrate determination.
          Approximately 50 ml of distilled water are added and the sample
          preserved with sulfuric acid for later analysis.  The procedure
          used for nitrates is the Modified Brucine Method as in Standard
          Methods, 12th Ed., pp 393~39^ or the automated method using a
          reduction column, sulfanilic acid and naphthylamine hydrochloride.

     5A  A portion of the wet sample is weighed for NHo-N and Org-N and
          approximately 50 ml of water are added with three or four drops
          of concentrated sulfuric acid for preservation and ammonia
          stabilization.  The sample is analyzed using the distillation
          method for ammonia nitrogen followed by a Kjeldahl digestion
          on the residue with subsequent neutralization and distillation.
          An aliquot of the distillate is nesslerized and read on a
          spectrophotometer and reported as Org-N.  NOTE:  Samples are
          distilled into 0.02 N H2SO^.

     5.5  A portion of the wet sample is weighed and percent total solids,
          volatile solids, silica, manganese, and iron are determined.
          Total solids are determined by drying the sample overnight at
          103°-105°C.  The fixed solids are determined by ignition in a
          muffle furnace at 600°C for an hour.  Volatile solids are
          determined by calculation.  A silica analysis is then made on
          the residue (Fixed Solids).  An aliquot of filtrate from the
          silica determination is analyzed for iron using the Phenanthroline
          Method described in Standard Methods, 12th Edition, pp 156-159.
          Another aliquot of the filtrate is analyzed for manganese using
          the Persulfate Method described in Standard Methods, 12th Edition
          pp U89-^92 with modifications as shown in procedures.

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Introduction to Bottom Sediment Analysis
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     5.6  A portion of the wet sample is weighed for determination of
          phosphorus (as P^.).  Due to the incomplete oxidation of the              •
          large amounts of organic matter present by the persulfate-               •
          sulfuric acid mixture,  a dry ash method using Mg (1^)3)2 '6H:0
          was found to accomplish complete oxidation.  After ashing,
          the Potassium Antimonyl Tartrate Method is used and the                  •
          resulting color development read on a spectrophotometer .                 •

     5-7  A portion of the wet sample is weighed for oil and grease                •
          analysis.  A procedure, which is essentially the same as in              |
          Standard Methods. 12th Edition, pp. 531-532, is employed.

     5.8  For special studies, a portion of the wet sample is weighed              •
          for long-term BOD determination in the same manner as the
          5-day BOD analysis.  The long-term BOD's are checked for DO
          content at various intervals over a period of 30 days   A DO             •
          analyzer is used to measure this parameter.  One-inch glass              ™
          rods are added to the DO bottle after each electrode insertion
          to return liquid level to a point where a water seal can be
          maintained upon cap replacement.  By the use of a DO analyzer,
          a long-term BOD analysis can be made using a single bottle
          throughout the entire period of 30 days .                                 •

     5.9  A portion of the wet sample is weighed for determination of              *
          cyanide.  The procedure used for cyanide is Colorimetric
          Method as in Standard Methods, 12th Ed., pp 1*55-457, with                •
          minor modifications as described in procedure.                           •

     5.10 A portion of the wet sample is weighed for determination of              •
          phenol.  The method used for phenol is essentially the same              |
          as in Standard Methods, 12th Ed., pp 517-519-

     5.11 A portion of the wet sample is weighed for determination of              •
          metals.  The procedures used for metals are new or modified              *
          methods developed by the Lake Michigan Basin Office - JWPCA.

     5.12 A portion of the wet sample is weighed for determination of              1
          sulfide.  The procedure used for sulfide is a modified method
          developed by the Lake Michigan Basin Office - PWPCA.                     •

     5.13 A portion of the wet sample is weighed for determination of
          chlorine demand.  The method used for chlorine demand is
          essentially the same as in Standard Methods, 12th Ed., pp 112-114.       •

6.   References
     Standard Methods for the Examination of Water and Wastewater,  12th ed.,
     APHA,  Inc.,  N.Y., 1965.
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               CHEMICAL OXYGEN DEMAND OF BOTTOM SEDIMENTS



1.   General Discussion

     (See Standard Methods, 12th Ed., p. £10-11, 196$)


2.   Apparatus

     Reflux apparatus, consisting of 250 ml Erlenmeyer flasks with ground-
     glass 2li/U° neck  and. 300 mm jacket Liebig, West, or equivalent
     condensers** with 2U/UO ground-glass joint, and a hot plate with
     sufficient power to produce at least 9 watts/sq. in. of heating
     surface, or equivalent, to insure an adequate boiling of the contents
     of the refluxing mixture«

3.   Reagents

     3.1  Sulfuric acid reagent, cone. H2SO^.

     3.2  Silver sulfate is dissolved in the HgSO,  acid.

     3.3   Mercuric sulfate, analytical grade  crystals


     3.1i  Ferroin indicator solution, 0.025 M:       Dissolve I.lj85 g
          1,10-phenanthroline monohydrate, together with  0.695 g FeSOi .
          7^0 in water and dilute to 100 ml.   This indicator solution
          may be purchased already prepared.


     3<>5  Standard potassium dichromate solution, 0.500N:   Dissolve
          120259 g I^CrpOy, primary standard grade, previously dried
          at 103°C for 2 hours, in distilled water and dilute to 1000 ml0


     3.6  Standard ferrous ammonium sulfate, 0.500N:  Dissolve 98 g
          Fe(NH^)2(SOj4)2.6H20 in distilled water.  Add 20 ml cone.
          H2^°li» cool and dilute to 1000 ml.  This  reagent must be
          standardized daily.

     3o7  Standardization of ferrous ammonium  sulfate: Dilute 25 ml
          standard potassium dichromate solution to approximately 250 ml.
          Add 50 ml cone. ^SOi  and allow to cool.   Titrate with the
          ferrous ammonium sulfate titrant,  using 2 or 3  drops of ferroin
          indicator.

               Normality - "*• K2Cr2°7 * "2*
                           ml
                *  Corning 5000 or  equal
               *#  Corning 2360,  9l5h8, or equal

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Chemical Oxygen Demand of Bottom Sediments 6
u.

















5.











6.



Procedure

U.I Add 25 ml K2Cr207 (0.2500N) to a suitable sized sample (0.5g-l«0g)of
bottom sediment that will consume one-half of the Kj^C^O^.
U.2 Add loO g HgSO^ and 0.75 g Ag^O^
Uo3 Add 50 ml of distilled water.
UoU Add 75 ml cone o KgSO^ (cautiously) and mix thoroughly.
U«5 Add boiling stones and reflux for 2 hours.
U.6 Cool and rinse condenser with distilled H20 (approximately
50 ml) while condenser is still connected to flask.
U«7 Disconnect and add approximately 150 ml distilled HpOo Cool
to room temperature and then titrate (after adding 3 to 5
drops ferrion indicator) to end point (the color changes
sharply from blue-green to reddish-brown).
U.8 For standard, use 25 ml K2Cr2Oy + 50 ml conc0 H2SO^ and dilute
to approximately 250-300 ml. Cool and titrate.
For blank,use 25 ml K2Cr2Oy, 75 ml H2SO^, 50 ml distilled
H20, 1 g HgSOjij, 0.75 g Ag2SOi|; reflux for 2 hours and continue
in the same manner as the sample.

Calculations
5.1 Wet basis
mg/kg« (a-b)c* x 8 x 1000
grams sample

5»2 Dry basis
mg/fcgs mg/Jcg wet basis
% solids

* a « mis of Fe(NH. )9(SCO2 required for blank
b - " " " M sample
c « normality of Fe(NH^)2(SO^)2
Reference
Standard Methods for the Examination of Water and Wastevater,
12th ed., APHA, Inc., N.Y., 1965, 510-511-

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                     CYANIDE IN BOTTOM SEDIMENTS


1. Scope


   Principle:  Simple cyanide salts will form hydrogen cyanide under
   acidic conditions.  Hydrogen cyanide has a boiling point of 26°C
   and can be easily isolated by distillation.  On the other hand, some
   complex cyanides are not easily decomposed and others, such as
   cobalticyanides are essentially non-responsive to this test.
   However, the conversion can be hastened by the addition of magnesium
   and mercuric salts.

2. Summary of Method

   The liberated cyanides are converted to sodium salts and reacted
   with chloramine T under slightly alkaline conditions to form cyanogen
   chloride.  A pyridine-pyrazolone reagent reacts with the cyanogen
   chloride to form a blue dye, the absorbance of which is measured at
   620 mn.  Increased sensitivity can be obtained by extracting the dye
   into n-butyl alcohol and reading the absorbance at 630 mji.


3. interference

   Most of the substances known or thought to interfere can be removed
   via the distillation procedure.  Common interferences are heavy metal
   ions, thiocyanates, cyanate, and other substances which may form cyanide.
   Sulfides and fatty acids interfere with the analysis and are not
   eliminated by distillation.  However, these materials are not often
   found in surface waters.  If they are known to be present, procedures
   for their removal are listed in the 12th  Edition Standard Methods. 1965.

U. Minimum Detectable Concentration

   The minimum detectable concentration is 0.02 mg/1 as cited in the
   12th Edition Standard Methods.  1965.
5. Apparatus


   5.1  Modified Claissen flask, 1,000 ml.


   5.2  Allihn condenser. (200 mm)


   5.3  Gas washer bottles (Sargent #39623) or equivalent.


   5.4  Suction flask, 500 ml.


   5.5  Water aspirator, air inlet tube.


   5-6  Heating element, for modified Claissen flask or bunsen burner.

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Cyanide in Bottom Sediments

     5.7  pH meter.                                                                 |

     5.8  Reaction tubes (test tubes).                                              •

     5.9  Spectrophotometer or filter photometer providing a 1 cm light
          path (or longer) and a wave length of 630 ntt.


 6.   Reagents

     6.1   Sulfuric  acid,  cone.                                                     •

     6.2   Sodium hydroxide,  IN: Dissolve k g NaOH in 100 ml of vater.

     6.3   Mercuric  chloride  solution:   Dissolve  3^ g of HgClg in 500 ml           •
           of distilled vater.

     6.k   Magnesium chloride solution:   Dissolve 51 B of MgClo^HgO in            I
           100 ml of distilled vater.

     6.5   Acetic acid, glacial.                                                    •

     6.6   Chloranine T, 1.0^ aqueous  solution.   (Prepare daily.  If
           reagent does not dissolve readily, degra
           is indicated and it should be replaced.)
      reagent does not  dissolve  readily,  degradation of the reagent            •
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6.7   l-phenyl-3-a-^nyl~5-pyrazolone,  saturated aqueous solution (7 g/l)       •
      (Eastman Reagent #1397).                                                  |

6.8   Bis-pyrazolone: Dissolve  0.025 g in 25 ml pyridine.  Prepare             _
      fresh daily.   (Eastman  reagent #6969).                                   I

6.9   Mixed reagent  immediately prior  to  use; nix 125 ml of reagent
      #6.7 vith 25 nl of reagent #6.8.                                         •

6.10  Stock cyanide  standard:   Dissolve 0.251 g KCII in one liter of
      distilled vater (add  two  pellets of sodiun hydroxide for                 •
      stability).  Solution contains 0.1  ag/ml cyanide.  Ihis is a             •
      primary standard and  must be  prepared, fresh each veel;.

6.11  working Standard:  Dilute stock standard 100 fold, 1 ml = 1 ^g.            •
       Take 0 - 5 ml  of the resulting solution for standard curve.                •
       Prepare fresh  daily.
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 Cyanide  in Bottom. Sediments



     6.12   5# disodium hydrogen phosphate.



     6.13  N-butyl alcohol - reagent grade.

     6.1k  The buffer solution is prepared by adding 35-0 ml 0.1 N.
           NaOH to 50.0 ml 0.1M KH2K>4 and diluting to 100 ml.  !J3iis
           buffer gives a pH of J.2.

           Kie water used in preparing both solutions should be free of
           carbon dioxide.  This can be accomplished fairly well by heating
           distilled water to boiling in a flask and cool while stoppered
           lightly.  If stopper is tight while water is hot, cooling may
           freeze the stopper.


7«   Procedure

     7«1   Distillation:   An  aliquot of a well-mixed bottom sediment sample,
           containing not more  than 500 mg of cyanide (lOg) is added to a
           Claissen flask and brought to approximately 500 ml  with distilled
           water.  Fifty  ml   of 1.0 N sodium hydroxide and 75 ml  of distilled
           water are added to the gas washer receiving vessel and the unit is
           assembled.  Air, drawn through the unit via vacuum aspiration, is
           adjusted so that its flow rate is approximately one bubble per second
           through the receiving unit, or as many as necessary to prevent flow
           back.


           After the air  inlet  tube is  positioned,  1.0 ml of mercuric chloride
           and 4.0 ml of nagnesium chloride solutions are added through the
           tube and two minutes are allowed for mixing. After rinsing the tube with
           distilled water,  5 ml  of  concentrated sulfuric acid are then added
           through the tube;  at this point, if it is not known vhcthar the pH is
           2.0 or lower,  the  pH should be determined on a separate yolame of the
           sample with the same amount of sulfuric acid added as in the distilled
           sample.


           Heat is applied to the system at a rate that will produce boiling,
           as soon as possible but will not allow a flow back.  Refluxing,
           which should begin within 30 minutes, is carried on for l£ hours.
           Occasionally   during distillation  the air flow rate may have to
           be readjusted  to prevent the reverse flowing of the sample solution.

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Cyanide in Bottom Sediments                                                          •

7.   Procedure (continued)                                                             _

    7.1  (continued)                                                                  *

         When the heating  is discontinued,  the air flow proceeds  for an              •
         additional 15 minutes for cooling  purposes.                                  |

    7.2  The contents of the gas washer vessel are drained into a 250 ml              M
         volumetric flask  and the accessible components of the distillation          •
         unit rinsed twice with distilled water.   The washings are added  to
         the distillate and the solution brought  to volume.

         Pipette a 15 ml aliquot-or other suitable volume- of the solution           •
         into a 25 ml volumetric flask.

         Neutralize to phenolphthalein end-point  using acetic acid (1:4 or           |
         weaker).

         Add 0.2 ml Chloramine-T, stopper,  mix, and after one minute, add            •
         2 ml of the buffer solution.                                                 ™

         Add 5 ml of the mixed pyrazolone,  dilute the solution to volume              I
         with distilled water, stopper, mix, and  allow to stand for 20 minutes       •
         for color development.

         The optical densities of the standard and samples are read against          •
         a reagent blank at 620 my on a spectrophotometer within one hour,
         at maximum.

    7.3  Alcohol extraction:  The sensitivity of detection is slightly               ™
         increased by determining the optical density of the colored dye
         in butyl alcohol.  Some interferences which may pass through the            •
         distillation may  also be eliminated.                                        |

         After color development in the aqueous system, add 2 ml  of disodium         _
         hydrogen phosphate solution and 10 ml of n-butyl alcohol.  Mix and          •
         withdraw an aliquot of the alcohol layer.  Measure optical density          *
         at 630 my.
7.4  Samples are to be compared with distilled standards,
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Cyanide in Bottom Sediments

8.     Calculation

       8.1  Wet Basis:

            mg/kg CN~ =  jSt

             9.
                                                      ?-D'        -,   x 1,000
                                                      (grains sample    '
                                                       in aliquot)
       8.2  Dry Basis

                  CN" =


       References
                                            vet basis
                                         Solids  (decimal fraction)
                    9.1  Standard Methods  for the Examination of Water and Wastevater,
                         12th  ed., APHA, Inc.,  N.Y.,  1965,
                    9.2  Ludzack,  et  al.,  Analytical Chemistry 26:  1?8^ ( 195*0 •

                    9.3  Jarrell,  R.,  Report,  Colorado River Basin  Project Laboratory.

                    9.4  Recovery  of  Cyanide by a Modified Surf ass  Distillation,
                         Charles T. KLly,  Unpublished.

                    9.5  Babcock,  R.,  American Oil Co.,  Unpublished Report.

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           TOTAL AND SOLUBLE FERROUS IRON IS BOTTOM SEDIMENTS
1.   General Discussion
2.2  Hot Plate.

2.3  Centrifuge and 50 ml tubes with caps.

2 A  Magnetic Stirrer .
                                                                                        I
     lol  Principle:  Iron is brought into solution and reduced to the                  |
          ferrous state by boiling with acid and hydroxylamine, and
          treated with 1,10-phenanthroline at pH 3.2-3.3.  Three                        _
          molecules of phenanthroline are required to chelate each atom                 •
          of ferrous iron to form an orange-red complex.  The colored                   •
          solution obeys Beer's law; its intensity is dependent upon
          pH and is stable for at least 6 months.  A pH between 2e9 and                 •
          3»lj insures rapid and maximum color development in the                        |
          presence of an excess of phenanthroline.

     1<>2  Interference:  Among the interfering substances are strong                    •
          oxidizing reagents, cyanide, nitrite, and phosphates - poly-                  •
          phosphates more so than orthophosphate; chromium; zinc in
          concentrations exceeding ten times that of iron; cobalt and                   •
          copper in excess of 5? mg/1, and nickel in excess of 2 mg/1.                   ||
          Bismuth, cadmium, mercury, molybdate, and silver precipitate
          phenanthroline.  The initial boiling with acid reverts polv-                  _
          phosphates to orthophosphate and removes cyanide and nitrite,                 •
          which would otherwise interfere.  The addition of more hvdroxyl-              •
          amine will eliminate errors caused bv excessive concentrations
          of strong oxidizing reagents.  In the presence of interfering                 •
          metal ions, a larger excess of phenanthroline is required to                  |
          replace that which is conplexed by these interferences.  With
          excessive concentrations of interfering metal ions, the
          extraction method is preferred.                                               •

          If much color or organic matter is present, it may be necessary
          to evaporate the sample, gently ash the residue, and then re-                 •
          dissolve in acid.  The ashing may be carried out in silica,                   |
          porcelain, or platinum crucibles which have previously been
          boiled for several hours in 1 + 1 HC1.                                        _

     1,3  A separate sediment sample must be taken for ferrous iron,  preserved          "
          with 12 ml cone. HC1 per 300 ml bottle, if sample is not
          preserved by freezing.                                                        •

2 .   Apparatus

     2.1  Colorimetric equipment:  One of the following is required:                     •

          a.  Spectrophotometer, for use at £10 j^ providing a light
              path of 1 inch or longer.                                                 •

          b.  Filter photometer, providing a light path of 1 inch or
              longer and equipped with a green filter having maximum
              transmittance near
                                                                                        I

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                                                                  13
Total and Soluble Ferrous Iron in Bottom Sediments



     2.5  Acid-washed glassware:  All glassware must be washed with
          cone. HC1 and rinsed with distilled water prior to use,  in
          order to remove the thin film of adsorbed iron oxide which
          is frequently present as a result of employing the glassware
          for other purposes.


3.   Reagents

     All reagents must be low in iron.  Iron-free distilled water is
     required.  Glass-stoppered bottles are recommended for storage.
     The hydrochloric acid, ammonium acetate solution, and stock iron
     solutions are stable indefinitely, if tightly stoppered.  The
     hydroxylamine and phenanthroline solutions are stable for several
     months.  The standard iron solutions are not stable and must be
     prepared fresh,   as needed, by diluting the stock solution.
     Visual standards in nessler tubes are stable for 3 months if pro-
     tected from light.

     3.1  85$ Phosphoric Acid.

     3.2  Hydrochloric Acid, cone,

     3.3  Hydrochloric Acid, 1/1 (oxygen free


     3.4  Hydroxylamine solution:  Dissolve 10 g N^OH.HCl in 100 ml
          distilled water.
     3»5  Ammonium acetate buffer solution:  Dissolve 2^0 g NHjjCpHoO,, in
          1^0 ml distilled water.  Add 700 ml glacial acetic acid and
          dilute to 1 liter.  (Since even good grade NHj^H-^ contains
          a significant amount of iron, new reference standards should
          be prepared with each buffer preparation.)


     3.6  Phenanthroline solution:  Dissolve 0.1 g 1,10 phenanthroline
          monohydrate, C^Ho^o^O in 100 ml distilled water by stirring
          and heating to 80°Cj do not boil.  Discard the solution if it
          darkens.  Heating is not necessary if 2 drops of cone. HC1 are
          added to the distilled water.  (Note that 1 ml of this reagent
          is sufficient for no more than 0.1 mg Fe.)


     3.7  Stock iron solution:  The metal (a) or the salt (b) may be used
          for the preparation of the stock solution, which contains
          0.200 mg Fe per 1000 ml0


          a.  Use electrolytic iron wire, or "iron wire for standardizing,"
              to prepare the solution.   If necessary,  clean the wire with
              fine sandpaper to remove  any oxide coating and  to produce a
              bright surface.  Weigh 0.2000 g wire and place in a 1-liter
              volumetric flask.  Dissolve in 20 ml 6N H2SOj1 and dilute  to
              the mark with iron-free distilled water.

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                                                                                     I
Total and Soluble Ferrous Iron in Bottom Sediments


          b.  If ferrous ammonium sulfate  is preferred,  add slowly
              20 ml cone. H2SO]L to $0 ml distilled water and dissolve
              l.UOU g Fe (NH^)2(SO|1)2.6H20.  Add dropwise 0.1N KMnO^                •
              until a faint pink color persists. Dilute with iron-                 •
              free distilled water to 1,000 ml and mix.

     3.8 Standard iron solutions:  These  should be  prepared the day                |
          they are used.

              Pipst £0.00 ml stock solution into a 1-liter volumetric               I
              £**1 f\ ev Ix nvNs»] rl ^ "1 1 1 4- rt 4- j-\ 4- Vl /•* W*^ •y«l*' Y_»n 4- V\ "I V»y-\Vl _ "f*T»*"»rt f^^ C* 4" ^ "1 H £\/3                 ^^*


                                                                                     I



     3.9 Ammonium hydroxide (l:l).

     3.10 Standard potassium dichromate solution, 0.25 normal (use                  •
          oxygen-free H20 ) .                                                          *

     3«11 Diphenylamine sulfonate indicator solution.  Prepare by                   I
          dissolving 3-17g of barium diphenylamine sulfonate in                     •
          distilled water and adding a solution of l.Tg  of sodium
          sulfate decahydrate (Na2SOj,.  • 10H20) to precipitate the                   •
          barium and dilute to one liter vith  oxygen-free H20.                      |

     3.12 Oxygen-free water.                                                         _

     3.13 Ferrous ammonium sulfate approximately 0.25N (use oxygen-free             ™
          a.
              flask and dilute to the mark with iron-free distilled
              waterj  1.00 ml = 10.0 HgFe.

          b.  Pipet 5.00 ml stock solution into a 1-liter volumetric
              flask and dilute to the mark with iron-free distilled
              water;  1.00 ml = l.OOu-g  Fe.
U.   Procedure - Total Iron                                                         I

     ii.l  Weigh 10 grams of a well-mixed sediment sample into kOO ml beaker
          and add 5 ml HN03, 085 ml H202 (302), plus 1 ml NaN03  (h%),               •
          and evaporated to drynesso                                                I

     li.2  Ash in muffle furnace at !?50°C for ^o minutes.                            •


     ii.3  Cool, add 1|0 ml HC1 (1:1), cover with watch glass, and heat
          gently for 20 minutes.                                                    _


     U.U  Allow to stand 5 minutes to cool (or longer) and filter into              *
          a 100 ml volumetric flask.


     U.3>  Dilute to volume, mix, and take 50 ml or an aliquot contain-              I
          ing 0.2^ rng iron (diluted to 50 ml) into a 100 ml volumetric
          flask.                                                                    •

     U.6  The sample is then made alkaline to phenophthalein (l drop)
          with ammonium hydroxide (1:1).

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                                                                     15

Total and Soluble Ferrous Iron in Bottom Sediments


     k.7  Add 2 ml HC1, 2 ml hydroxylamine hydrochloride  solution  and boil,
          reducing the volume to approximately 20 ml.


     k.8  Add 10 ml  acetate buffer*, 10 ml phenanthroline solution and
          dilute to  100 ml.  Mix thoroughly and  allow 15  minutes for
          maximum color development.  Read adsorbance against  a blank
          at a wave  length of 510 n^ on a spectrophotometer.
     4.9  Calculations are as shown in"6" below.

Jj.   Procedure - Soluble Ferrous Iron


     5.1  Weigh 10 grams of a well-mixed sediment sample  into  a 250 ml
          beaker, add 100 ml  of oxygen-free distilled water and digest
          on hot plate for 30 minutes.


     5.2  Filter into a 100 ml volumetric flask.


     5.3  Dilute to  volume, mix and take 50 ml or an aliquot containing
          0.25 mg of iron (diluted to 50 ml) into a 100 ml volumetric
          flask.


     5.4  Then proceed as in total iron sections 4.6 and  k.8


6.   Calculation
     6.1  Wet basis
                               O.D. Sample
                  O.D. Std.    (grams sample
                                in aliquot)
     6.2  Dry basis

               _ mg/g wet basis
                   solids (decimal fraction)
* Check pH of solution sample on pH meter.  Range should be 2.9-3.!*-
  units for good color development.

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                                                                                   I
Total and Soluble Ferrous Iron in Bottom Sediments
     Procedure - Total Ferrous Iron by Titration
                                                                                   I
         Weigh an 8-10 gram sample of veil-mixed sediment in a                     |
         150 ml beaker and add sufficient hydrochloric acid to
         extract the expected iron concentration, 10 ml of 85                      «
         percent phosphoric acid, plus enough oxygen-free water                    •
         to bring total volume to approximately 50 ml.

         (a)  For each mg of total iron expected add 1.2                           •
              mg of HC1 plus a 10 percent excess.  (20 ml  1/1 HCl)              I

         (la)  In moderately dilute solution and at room temp-                      •
              erature, dichromate does not react with chloride ion.                •

         (e)  Phosphoric acid is added for two reasons:

                   To complex any ferric ion initially present in                  •
                   the specimen, thus preventing its reduction to
                   the ferrous state by hydrogen sulf ide during                    •
                   extraction .                                                     |

               (2)  To prevent the indicator from changing color                    •
                   before reaching equivalence point by increasing                 •
                   oxidation potential to —  .85.

     7«2 Boil on a hot plate for 15 minutes.  Take care to avoid                   I
         frothing by adding more deoxygenated water or by removing                 •
         from heat and stirring.

         Note:  Volume must be 50 mis or less at conclusion of                     |
                15-minute boiling period.

     7-3 Transfer and rinse with small aliquots of deoxygenated water              •
         into a 50 ml centrifuge tube with cap and centrifuge for                  ™
         10 minutes .

     7.4 Decant solution into a  500 ml Erlenmeyer flask containing                 8
         approximately 200 ml of oxygen-free water.  Rinse centri-
         fuge tube with small aliquots of water.                                   •

     7.5 Add 1 ml of indicator solution.  Titrate with standard
         dichromate until the first permanent coloration appears.
                                                                                   I

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                                                                                 17
             Total and. Soluble Ferrous  Iron in Bottom Sediments
            8.    Calculation
                                         * C x 33.9
                                                    X 1000
 I                    Iron,  mg/kg = *—-   D
 «                    A • milliliters KpCr_07 solution required to titrate sample
 *                    B = milliliters K2Cr20  to titrate blank
 8                    C = normality of K2Cr20
                      D = grams sample used (dry basis)
 •          9«  References
 _              9«1   Patterson, A., Thomas, H.C., Quantitative Analysis,  Henry Holt
 •                    and Co.,  New York 1952,  pp 278-287.
                9-2   Fisher,  R.B., Quantitative Chemical Analysis, W.B. Saunders Co.,
 •                    Philadelphia, 1957,  PP 28^-5.
                9-3  Standard Methods for the Examination of Water and  Wastewater,
 •                   12th ed.,  APHA, Inc.,  N.Y.,  1965,  156-159.

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                                                                                    I

                     METALS IN BOTTOM SEDIMENTS                                     •

                          Atomic Absorption
1.   General Discussion                                                             •

     Metal complexes are freed by digestion of bottom sediment samples
     with concentrated HNO^ acid H202 (30$).  A wide range of elements
     can be determined by atomic absorption with the selection of the               •
     proper source (hollow cathode lamp).                                           •

     1.1  Principle:  The principle of atomic absorption and its application        •
          to the analysis of metals is based upon the element of interest           |
          being dissociated from its chemical bonds and placed into an
          unexcited, un-ionized "ground" state.  It is then capable of              •
          absorbing at discrete lines of narrow band width radiation                •
          provided by a hollow cathode lamp with a cathode made of the
          element being sought.  The absorption of the light by the metal
          in the ground state is related to the concentration of the metal          I
          being sought.                                                             I

     1.2  Interference:  The atomic absorption method is relatively free            •
          of interferences.                                                         •

2.   Apparatus                                                                      _

     2.1  Atomic absorption spectrophotometer.                                      "

     2.2  Muffle furnace.                                                            •

     2.3  Pipettes.

     2.4  250 ml beakers.                                                            j|

     2.5  Watch glasses.

     2.6  250 ml volumetric flasks-                                                 •

     2.7  Polyethylene bottles (60 ml).                                             •

     2.8  Centrifuge (20,000 rpm).

3-   Reagents                                                                       •

     3-1  Nitric acid (cone, metal-free).

     3.2  Hydrogen peroxide (30$).                                                  •

     3.3  Hydrochloric acid (cone, metal-free)•
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                                                                  19


Metals in Bottom Sediments- Atomic Absorption


     3-4  Ammonium chloride.


     3-5  Calcium nitrate.

     3.6  Redistilled water or double deionized water  (metal-free).


     3-7  Bottled acetylene gas.


     3-8  Bottled nitrous oxide gas.


     3 '9  Compressed air.


it-.   Preparation of Standards


     4.1  Using a commercial certified atomic absorption standard,
          containing 1,000 mg/1 of the metal, prepare  an appropriate
          dilute stock standard.  Standards may be prepared in the
          laboratory using pure metals .


     U.2  Make up working standards to approximate the concentration
          in the sample using the dilute stock standard .


5 •   Preparation of Samples

     5-1  Place 2.5 g    of a well mixed bottom sediment sample in a
          250 ml beaker, add 10 ml of cone. MO- acid  and 0-5 ml of
               (30$) and evaporate to dryness.
     5-2  Ash at UOO-lt-25°C for one hour in a muffle  furnace and cool.


     5-3  Add 25 ml of acid mixture  (200 ml of cone. HNO^>, 50 ml cone. HC1
          and 750 ml of redistilled  water), 20 ml of lO^NH^ci and 1 ml
          of Ca (N0_,)2 • UH20 (11.8   g/100 ml).  Heat gently for 15 minutes
          and cool for five minutes  or longer.


     5.^  Transfer sample to centrifuge tube and centrifuge for 10 minutes
          at 20,000 rpm.  Transfer the supernatant to a 250 ml volumetric
          flask.  Rinse the residue  in the centrifuge tube twice with
          redistilled water and add  washings to supernatant and dilute to
          volume.  Then transfer sample to a small plastic bottle.


6.   Method for Cu, Cd, Ni, Zn, Cr ,  Pb, Mn, Al and Other Metals


     Follow procedures as outlined in manufacturer's manual.

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                                                                   20
Metals in Bottom Sediments -Atomic  Absorption
8.   Atomic absorption is the preferred method because  it  is much  faster,
     more  accurate  and there are less  interferences.
                                                                                    I
                                                                                    I
7-   Calculations                                                                   •

     7-1  Wet Basis

          By use of table from manufacturer's manual, convert percent               •
          absorption to absorbance.


          mg/kg - -~-—	—'——7~r~ X dil. factor X absorbance of sample X         •
           ^      absorbance of std.                                   *

                  1,000	                                                    •
                  gof sample/L                                                  I
     7-2  Dry Basis

             I    . mg/kg vet basis	                                       •
          mg/ s  " $ solids (decimal fraction                                       •


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                                                                      21
                           METALS IN BOTTOM SEDIMENTS
                                Polarographically

1.   General Discussion
       'sediment
     Metal complexes are freed by digestion of /   with HNOo acid.
     Simultaneous determination of copper, cadmium, nickel, and zinc
     is possible, also a v/ide range of other elements can be determined
     polarographically with the selection of the proper supporting
     electrolyte.

     1.1  Principle:  Cations are reduced on the surface of a dropping
          mercury electrode to form an amalgam in a current producing
          reaction.  This current is reflected in a galvanometer and
          may be recorded as a measurable wave which is proportional to
          the concentration of a specific cation that is being reduced
          at its specific half-wave potential.  The wave height produced
          by an unknown is compared to that made by a known standard
          for determining the amount of an unknown metal in the sample.

     1.2  Interference:  Any volatile or organic interference is
          eliminated by the ashing process in which carbon is destroyed.
          In addition, iron, chromium, turbidity,  chlorides, oxidizing,
          and reducing substances, and dissolved oxygen are major inter-
          ferences that are likely to be encountered,  The following
          procedures are designed to eliminate or reduce to a minimum
          these types of interferences.

     1.3  Precautions:  The one most precautionary measure to take is
          the washing of all glassware and other containers with hot
          HC1 (l:l) and rinse with distilled water.  Etched beakers
          should be avoided since they could possibly retain metals.
          Any wasted mercury should be removed to  prevent the poisonous
          fumes from volatilizing into the air.

2 o   Apparatus

     2.1  Sargent Model XV Recording ^olarograph
     2.2  MicroRange Extender
     2.3  Muffle Furnace
     2.1i  Pipettes (Kohr and Volumetric)
     2.^  600 ml Beakers
     2.6  Watch Glasses (to cover 600 ml beakers)
     2.7  10 ml Beakers
     2.8  Hot Plate
     2.9  ^O ml Volumetric Flask
     2.10 Folded Filter Paper (Whatman #12,  18.$ cm or equivalent)
     2.11 Polyethylene Bottles  (60 ml,  2 liter,  and wash  bottles)

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                                                                       22          •
Metals in Bottom  Sediments -Polarograpihically


3.   _Reagents (Not mentioned in procedures)

     3.1  Mercury (high-grade, metal- free)                                         •
     3e2  Nitric Acid (cone., metal-free)                                          •
     3.3  Hydrochloric Acid (conce, metal -free)
     3oU  Redistilled Water (metal-free, stored in polyethylene bottles)           •
     3.5  Sodium Sulfite, reagent grade                                            |
     3.6  Bottled Nitrogen Gas (water-pumped)

U.   Preparation of Standards                                                      •

     li.l  A stock standard solution io  made by dissolving the salts of            _
          the metals in water or the metals in nitric acid in such                 •
          proportions that 1.0 ml of standard equals 00010 ng of each              ™
          of the six metals (Cu, Cd, Ni, Zn, Pb, and Cr).

     U.2  A working standard is  prepared by diluting 100 ml of stock              I
          standard to 1 liter.  £.0 ml contains 0.005 mg of each of the
          metals .                                                                  •

     U.3  A 5.0 ml portion of the working standard  is  pipetted into a
          10 ml beaker and evaporated to dryness.  This standard is                H
          treated exactly as the samples and the comparative wave heights          •
          are  used in calculating the amount "of the particular metal              *
          in the samples.
                            Sediment                                               I
5«   Preparation of Bottom /   Samples for Metals on the Polarograph               •

     5.1  To a 10 gram sample of well-mixed bottom mud in a 600 ml beaker,         •
          add 10 ml HNO-j (cone.) and 0.5 ml K202 (3Q3) and evaporate to            |
          dryness .(If sample is not very uniform, use 20 grams.)

     £.2  Ash at Ii50-500o for 30 minutes in muffle furnace and cool.               I
                                      ml                                           •
     5.3  Add 25 ml acid mixture , 200/HNO-j cone. + 50 ml HC1 cone.  +
          750 ml redistilled water ,  heat gently 15 minutes and cool               I
          5 minutes or longer.                                                     •

     5.1  Filter and dilute the filtrate to 50 ml in volumetric flask,             •
          then transfer to a snail plastic sample bottle.                          |

     5»5  Pipette three 5 ml portions into 10 ml beakers and evaporate             •
          to dryness for analysis .                                                 •

6.   Method for Cu, Cd, Ni, and Zn on the Polarograph

     6-1  Reagents:   Prepare all reagents in metal-free distilled water.            •
      a.  Electrolyte:   Dissolve 10? grams  of NHrCl,  ll.li grams  (NHrJpCO^,          •
          and 6? ml NHj|(OH)2 in distilled water.   Add 10 ml of 0,2%                I
          trlton-X and  dilute to 1 liter.  The strength  of this  solution
          is 2. CM NH^Cl, OclM (NHj^gCO^,  and 1M NH^OH.                             •

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                                                                     23
Metals in Bottom Sediments - Polarographically
      b.   Sulfuric Acid (l:£).

      c.   Sodium Sulfite Solution (saturated)

      d.   Water-pumped Nitrogen Gas (This gas passed through a saturated
          solution of sodium sulfite just prior to use.)

 6,2  Procedure
•                 a.   To the dried sample or standard,  add 2  drops H^SO^'L:^)  and
          heat gently until near dryness.

     b.   Heat strongly until fumes cease.

     c«   Cool, add £«0 ml electrolyte and allow 5 minutes contact time,

     d.   Police the beaker and transfer sample to cell.

     e.   Add 5 drops sodium sulfite solution and allow nitrogen gas*
          to flow through cell for one minute.
                   f.    Add Mercury pool  and,  if  any  air  bubbles are present, swirl
•                      gently to remove,,

                   gt    Record.polarogram.

I                 h.    Recommended settings:  Sensitivity  0.002 (if range extender
B                      is available,  otherwise use 00003), voltage range 0 to -2.
                        Compensation and  damping  should not be used unless necessary.

         7.   Fethod for Lead on the Polarograph

I                   The normal interference  of  iron is minimized by the use of hydro-
                   xylamine hydrochloride in this method.

H            7.1  Reagents

•                 a.   £-—  Electrolyte - 0.1M KC1

I                 b«        Hydroxylamine-hydrochloride -

                   Co        Hydrochloric  acid  - 1:25
      cJ0       V/ater-pumped nitrogen gas (passed through saturated
              sodium sulfite just prior to use)
     *- The nitrogen gas is first washed by passing through a
       flask containing sodium sulfite (saturated).

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Metals in Bottom Sediments . Polarographically
7,2  Procedure
                                                                                        I

          a.   To the dried sample or standard, add 1.0 ml KG1 (1:25) and
               heat gentlv for about 3 minutes (heat must be such that                  •
               solution does not go to dryness ) .                                        •

          b.   Add 2 drops hydroxylamine-hydrochloride solution and heat
               3 more minutes (again solution must not go to dryness),,  If              I
               a yellowish color due to high iron content persists, add                 I
               hydroxylamine hydrochloride dropwise until the color disappears
               and heat for 1 more minute.                                              •

          c.   Remove, cool, and add 3>»0 ml electrolyte.

          d.   Police, transfer to cell, and allow nitrogen gas to flow                 I
               through sample for £ minutes.                                            •

          e.   Add mercury pool and record polarograin, using nitrogen blanket.          •

          f.   Recommended settings:  Sensitivity -0.001, compensation and
               damping as needed.

 8.   Method for Total Chromium (10)                                                     I

     This method is based upon the oxidation of chromium by alkalibromite               •
     and the simultaneous precipitation of the normally interfering iron,               I

     8 .1  Reagents :

          a.   Oxidizing solution - 100 ml Oef>?T NaOH solution mixed with                •
               £ ml saturated bromine water.

          b.   Electrolyte - 0.2N NaOH solution am5 0.1 g gelatin/liter.                |

          c.   Sodium sulfite - saturated anhydrous solution.

          d.   Nitrogen gas - water pumped.                                             I

     B <>2  Procedure                                                                     •

          a.   To the dried sample or standard, add O.U ml oxidizing
               solution and heat for 2 minutes on hot plate just below
               boiling o                                                                 I

          bo   Remove and allow to cool for £ minutes or longer.

          c.   Add £.0 ml electrolyte and transfer to cell, policing.                   •

          d.   Add 2 drops sodium sulfite solution and allow nitrogen gas
               to flow through sample for 5 minutes.                                    •

          e.   Add mercury pool and record polarogram, using nitrogen
               blanket .                                                                 •

          fc   Recommended settings: Sensitivitv -0.003, compensation -0,
               damping -0.
                                                                                        I

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•    Metals in Bottom Sediments - Polarographlcally
                                     •bottom sediments
      9*   Calculation of Metals in /    (or any solid material) in mg/kg
•         9.1  Wet Basis:
                mg/kg c ^ S. sample           H* sample	x O.OC£  xl	x 10oo
                         S. standard         H« standard           1    (grams sample
I                                                                       in aliquot)
•              where S » sensitivity
•                    H * wave height in mm
                  0.005> B mg metal in 5 ml std. (assuming £ ml std. and sample
I                          are used; this figure is changed if different amounts
                          of std. are used)

I
•               solids = decimal fraction of solids in wet sediment
                Example
I              wet basis
                                0.005   x  g  x  C^  x,	-^X1000
 _                              0.002      25        1     (grams sample
 I                                                         in aliquot
 "         9.2  Dry Basis:
                |mS/kgc ,^g/^gT/?et basis
                        % solids(deciroal fraction)
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                                                                                    I
 Metals in Bottom Sediments - Polarographically
                              REFERENCES
I

I
 1.  Report of the Advisory Committee on Revision of the Public Health
     Service 19^6 Drinking Water Standards.  (Mimeo.)                               _

 2.  Doudoroff, P. and Katz, M.  Critical Review of Literature on the               ™
     Toxicity of Industrial Wastes and Their Components to Pish.  II.
     The Metals, as Salts.  Sewage and Industrial Wastes, 2J5, 802 (1953).           •

 3.  Kroner, R. C., Ballinger,  D. G., and Kramer, H.P.  Evaluation of
     Laboratory Methods for Analysis of Heavy Metals in Water.  Journal             •
     of American Water Works Association, ;?2, 117 (i960).                           •

 k.  Love,  S. K.  Analytical Instruments Used in the Modern Water Plant
     Laboratory.  Journal of American Water Works Association, fr3, 731 (1951).      •

 5.  Wyatt, J. W.  Instrumentation in the Laboratory.  Journal of American
     Waterworks Association, *£, !38 (1953).                                       •

 6.  Butts, P. G., and Mellon,  M. G.  Polarographic Determination of Metals
     in Industrial Wastes.  Sewage and Industrial Wastes, 23, 59 (l95l)»            •

 7.  Lingane, J. J.  Systematic Polarographic Metal Analysis Characteristics        *
     of Arsenic, Antimony, Bismuth, Tin, Lead, Cadmium, Zinc, and Copper, in
     Various Supporting Electrolytes.  Industrial and Engineering Chemical          I
     Analysis.  Ed. 1^, 583 (19^3).                                                 •
 8.  Manzel, R. C., and Jackson, M. L.  Determination of Copper and Zinc            •
     in Soils and Plants.  Analytical Chemistry, 23, l86l (l95l).                   |

 9.  Lingane, J. J., and Kolthoff, I. M,  Polarographic Study of the Reaction
     of Chromate Ion at the Dropping Mercury Electrode.  Journal of American        I
     Chemical Society, 62, 852 (19^0).                                              •

10.  Standard Method for Polarographic Determination of Lead and Chromium           •
     in Zinc.  ASTME-68-56.  American Society of Testing Materials.                 |

11.  Hawkins, R. C., and Thode, H. G.  Polarographic Determination of               •
     Copper, Lead, and Cadmium in High Purity Zinc Alloys.  Industrial and          •
     Engineering Chemical Analysis, Ed. 16, 71
                                                                                    I

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 |            Metals  in  Bottom Sediments  - Polarographically


 I                                        REFERENCES  (continued)


              112.  McFarren, E.  F.,  Campbell, J.  E., and Engle,  J.  B.   Variations in
                  Zinc and  Copper  Content of Bivalves,   (in Press).

              113.  Official  Methods  of Analysis of  the Association  of  Official
                  Agricultural  Chemists,  Ninth Edition,  page  333,  (i960).

              14.  Standard  Methods  for the Examination  of Water and Wastewater,
 •                12th ed., APHA, Inc., N.Y., 1965, 458-466.



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            AMMONIA HITROGEN IN BOTTOM SEDIMBIIS  - DISTILLATION                     •


1.   General Discussion                                                            ™

     1.1  Principle:  Free ammonia nitrogen can "be recovered by                    •
          distillation of a sample at pH 7A.  Since natural waters                •
          exhibit varying pH values and buffering properties, a phos-
          phate buffer is applied to maintain the required pH during               •
          the distillation process.  The free ammonia distillate is                I
          collected in boric or sulfuric acid solutions to minimize
          ammonia losses.                                                          •

          Samples containing high ammonia concentrations (5-10 mg/l)
          should be analyzed following the procedure described in                  _
          "Standard Methods for the Examination of Water and Wastewater"           I
          12th 2d., 1965, Method A, page 191, Sec. k.k.                            U

          The concentration of ammonia in the distillate det^.Tnines                •
          the final method for neasurement.                                        •

          If the sample contains from 0.01 to 0.2 icg/1 ammonia                     •
          nitrogen, the distillate from 500 ml of sample is collected              •
          in 0.02N  H2SOj,.  The acid solution is concentrated on a
          steam bath to 5° ml producing a 10:1 concentration.  The                 •
          ammonia in the concentrated solution is then reacted vith                I
          Kessler's reagent to form a characteristic yellow-brown
          color vhich is measured at a wave length of ^25 xr^i.                      •

          Samples vhich contain from 0.2 to 1.0 mg/1 are distilled
          Into dilute boric acid solution.  The acid distillates                   _
          are diluted to the original 500 ml volume and a 50 ml                    •
          aliquot is taken for Nesslerization as outlined above.

          Samples that contain more than 1.0 mg/1 arrjronia nitrogen                 •
          are distilled iaio boric acid and can then be titrated                   •
          vith standard sulfuric acid using an appropriate indicator.

     1.2  Interference:  Ammonia recovery will be low on water samples              |
          containing nore than 250 mg/1 calcium unless the treatment
          prescribed in Sec. k.2 is followed.  The calcium and the                 •
          phosphate buffer react to precipitate calcium phosphate,                  |
          releasing hydrogen ions and lowering the pH.  A number of
          aliphatic and aromatic amines,  organic chloramines,  acetone,              _
          aldehydes, and alcohols,  and other undefined organic                     I
          compounds, cause trouble in direct nesslerization.  Compounds
          of this type have been found to yield a yellowish or greenish
          off color or a turbidity following the addition of nessler               I
          •ycpcrpn-f-. t.n fl-i c-M 11 n"hp<; on! T pr-hpfl frnm phlnrinat.pd sarvnl es .               •
reagent to distillates collected from chlorinated samples,
                                                                                   I

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                                                                29

Ammonia Nitrogen in Bottom Sediments - Distillation
         tChe titration procedure  is  also subject  to  amine  inter~
         ference because  the  standard  acid  can  react with  such
         alkaline bodies.  Eovever,  the  titration procedure  is
         free of interference from neutral  organic compounds.
         Sulfid.e Ms elso been reported  to  cause  turbidity
         following nesslerization, a condition  which may be
         avoided by adding  lead ce.rbona.te to  the  flask prior
         to distillation.  Volatile  substances  such  as formal-
         dehyde can be removed by boiling at  low  pH, after which
         the scciple GCJI be  distilled and nesslerized in the
         normal panne r .

2.  Apparatus

    2.1  Distillation apparatus:  A glass flask of 800 ml capacity
         attached to a vertical condenser is so arranged that the
         distillate falls directly into the collecting glassware.

    2.2  Digestion apparatus:  Provided with a suction take-off
         to remove water vapor and sulfur trioxide fumes .

    2.3  Colorimetric equipment:  A spectrophotometer for use at
             to 425 m, providing a light path of 1  inch or  longer.
    2.4  Kessler tubes:  Matched 50 ml* tall forms.

3.  Reagents

    3.1  Ammonia-free water.

    3.2  Phosphate Buffer Solution pH 7.4:  Dissolve l4.3 g
         anhydrous potassium dihydrogen phosphate,  KH2POlj., and
         68.8 g   anhydrous dipotassium hydrogen phosphate
         and dilute to 1 liter with ammonia-free water.

    3.3  Boric Acid Solution:   Dissolve 20 g   anhydrous boric
         acid HoBO^ in ammonia-free water and dilute to 1 liter,

    $.k  Sodium Hydroxide-Sodium Thiosulfate reagent:   Dissolve
         500 g   NaOH and 25 g   ITa2S2Oo • 5H20 in  ammonia-free
         vater and dilute to 1 liter.

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                                                                                   I
Ammonia Nitrogen in Bottom Sediments - Distillation


    3«5  Phenalphthalein Indicator Solution:  Dissolve 5 £
         phenalphthalein in 500 ml 95 per cent ethyl alcohol
         and add 500 ml distilled water.   Then add 0.02N NaOH                      •
         dropwise until a faint pink color appears.                                •

    3.6  Mixed Indicator:  Mix 2 volumes  of 0.2 per cent                           •
         methyl red in 95$ alcohol with 1 volume of 0.2 per cent                   |
         methylene blue in 95 per cent ethyl alcohol.   This
         solution must be made fresh every 30 days.                                •

    3-7  Standard Sulfuric Acid Titrant 0.02N:  In this strength
         1.00 ml equals 0.28 mg N.

    3.8  Kessler reagent:  Dissolve 100 g   mercuric iodide,  Hglp                   •
         and 70 g:   potassium iodide,  KI,  in a small quantity of
         ammonia-free water, and add this  mixture slowly, with                     •
         stirring,  to a cool solution of  160 g:  NaOH in §00 ml                     I
         ammonia-free water.  Dilute to 1  liter and store in dark
          ?yrex bottle out of sunlight. Stable about one year.                      •
          Caution Toxic)                                                           •

    3.9  Standard Ammonium Solution:   Dissolve 3.8l9 g   anhydrous
         ammonium chloride,  NH- Cl,  dried at 100°C in ammonia free                   •
         vater and dilute to 1,000 ml: 1 ml = 1.0 mg N-(1022 mg NH^).              •
         Then dilute 10.0 ml stock ammonium chloride solution to
         1,000 ml with ammonia-free water.   1 ml = 10.00 ng N                      •
       (!2.2MgNH3).                                                             I

    3.10  standard Organic Nitrogen Solution:   Dissolve 1.0503 g-                    _
         of glutamic acid dried at 100°C in ammonia-free water and                  I
         dilute to 1,000 ml = 1 ml = 0.1 mg N.
Procedure

U.I  Add 500 ml distilled water, 10 ml phosphate buffer solution
                                                                                   I
         and a few boiling chips  to  an 800 ml flask and stean put                   •
         the entire distillation  apparatus until the distillate shows               |
         no  trace  of ammonia.

      2  Place a previously prepared add sample in" a                               |
         800 ml kjeldahl  flask and add 500 ml of ammonia-free distilled
         vater. * Neutralize to about pH 6.6 using a pH meter if
         necessary then add 10 ml phosphate buffer solution.  Add a                 I
         few boiling chips.                                                        ™

      Boil to remove H?S, if present.                                              •
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31
Ammonia Nitrogen in Bottom Sediments - Distillation
4.3 Distillation: Distill over 300 ml at the rate of 6-10 ml/min
collecting the distillate in 50 ml of -02N H2S01»..* Place
distillate in a glass stoppered 500 ml graduate cylinder and
bring up to $00 ml with ammonia free water and mix.
U.lt- Color Development: Place 50 nil portion of the distillate in
a 50 ml nessler tube. Add 1 ml of nessler reagent and mix
thoroughly by inverting the tube Blx times. Allow to stand
20 minutes and read at ^25 mp. .
4.5 A standard and blank should also be run in the same manner
as the sample.
5. Calculation
5.1 Wet Basis:
mg/fcg= u\g in Std. x O.D. Sample x 1000
O.D. Stdo ( g Sample
p n -n. T> .. in aliquot)
5.2 Dry Basis: H
mD./, - mg/k-K wet basis
1"e""v ' % Solids (decimal fraction)
*Boil to remove HpS if present.
6 . Reference

Standard Methods for the Examination of Water and Wastevater,
12th ed., APHA, Inc., N.Y., 1965, 191.

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                                                                                  I
     NITRATE AND NITRITE NITROGEN IN BOTTOM SEDIMENTS-AUTOANALYZER


1.  General Discussion                                                            •

    1.1  Principle:  The nitrate in bottom sediment is reduced to                 •
         nitrite by passing the sample stream through a ten-inch                  I
         column of zinc granules (20 mesh) in the presence  of sodium
         acetate and hydrochloric acid buffer.  The hydrogen gas is               •
         allowed to escape through a T vent.  The sample stream is then           J
         joined by the color reagent where the nitrite is determined by
         the formation of a reddish purple azo dye, at a pH 2.0 to 2.5,           .
         by coupling of diazotized sulfanilic acid with naphthylamine             •
         hydrochloride.  The stream then passes through a delay column            ™
         to allow more time for color development, and then through the
         colorimeter where the optical density is measured at a wave              •
         length of 520 millimicrons.                                              •
    1.2  Interference:  See Standard Methods, 12th Edition, 1965,
         page 205.

    1.3  Minimum Detectable Concentration:  0.01 ing per liter can be
         ru.HJ.muiu .ut;i,t:c:i/a.u>j.c ounccin/i a.i»j.uu;  v.wx ing ±>cr J.XUCM cttfi  ue              •
         detected with a reasonable degree of precision and accuracy.             I


2.  Apparatus
    Technicon Auto-Analyzer, consisting of large sampler, proportioning
    pump, colorimeter and recorder. See flow diagram in Figure 1.

3-  Reagents

    3.1  Sodium acetate solution M/4:  Dissolve 3^ g NaCpH 0  »3E0 in
         distilled water and dilute to 1,000 ml.          ^
         and naphthylamine hydrochloride reagent.  Protect from light
         and refrigerate to increase stability.
                                                                                  I
    3.2  Sodium acetate hydrochloric acid buffer solution:  Mix 100 ml
         of HC1 (1/99) with 1,000 ml of sodium acetate solution M/U.            •

    3.3  Sulfanilic acid reagent:  Completely dissolve 3.0 g of
         Bulfenilic acid in 350 ml hot distilled water, cool, add 100 ml          •
         concentrated hydrochloric acid,  dilute to 500 ml with distilled          |
         vater and mix thoroughly.

    3.4  Naphthylamine hydrochloride reagent:  Dissolve 3.0 g naphthyl-           I
         amine hydrochloride in a solution containing 150 ml distilled
         vater, 5 ml concentrated hydrochloric ecid and 250 ml of 95$             m
         ethyl alcohol.  Dilute to 500 ml with distilled water and mix            I
         thoroughly.                                                              ™

    3.5  Color reagent:  Mix equal volumes of sulfanilic acid reagent             •
                                                                                   I

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                                                                      33

Nitrate and Nitrite Nitrogen in Bottom Sediments- Auto-Analyzer


    3.6  Zinc metal:  20 mesh granular reagent grade; vash with
         1/99 HC1, rinse with distilled water followed with a
         rinse of ethyl alcohol, dry and rinse with chloroform.

    3.7  Stock nitrate solution:  Dissolve J.2180 g anhydrous KNO~
         in distilled water and dilute to 1,000 ml.  This solution
         contains 1.00 mg of nitrate nitrogen per ml.

    3.8  Lower working standards are prepared from the stock solution.
         Concentrations dependent upon anticipated levels to be found
         in sample.

U.  Procedure
Place a previously prepared sample into a 200 ml pyrex
volumetric flask and add distilled HpO to approximately half-
                                                     SO, .
                                             p
         fill volumetric flask and add 5 or 6 drops of cone.
    k.2  Boil for 15 minutes to dissolve all nitrates from the sediment
         and cool.

    ^.3  Centrifuge at 2,000 RPM for 5 or 10 minutes, then decant into
         volumetric flask.  Wash residue with distilled water by
         thoroughly mixing residue with a stirring rod, follow by
         centrifuging .

    k.fy  Decant wash solution into volumetric flask and repeat a second
         time.

    U.5  Make up to volume with distilled water.

    lj.6  Filter aliquot through membrane filter (having a porosity
         of O.^5f0   into test tube.

    k.f  Place samples in Technicon Auto- Analyzer sampler with one
         distilled water wash between each sample.

    b.8  Permit 3° minutes for instrument warm-up and establish a
         base line.

    H.9  Set the instrument for two minute sampling  time and slightly
         greater than two minutes  for relay time.

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                                                                  34


Nitrate and Nitrite Nitrogen in Bottom Sediments- Auto-Analyzer                   •


5-  Calculation                                                                   •

    5.1  Mg/1 in aliquot may be calculated by comparing peak height
         of sample with standards, employing a chart reader.

                      '"g/1 in aliquot _        x Dil. Factor  x 1000          •
                      g/L of sample in sample" vol."
          dry basis:                                                              •
                                                                                  I
                                basts) _
                        solids (decimal fraction)

         Nitrite may be determined on the auto-analyzer by by-passing              |
         the zinc column and manually according to Standard Methods,
         12th Edition, 1965, pages 205-208.                                        .

6.  References

    6.1  James E. O'Brien and Janece Fiore Dio of Laboratories and                 •
         Research, New York State Department of Health                             "

    6.2  Standard Methods  for the  Examination of Water  and Wastevater,              •
         12th ed.,  APHA,  Inc., N.Y.,  1965, 205-208.

    6.3   L. J. Kamphake,  Robert A. Taft Sanitary Engineering Center,
         Cincinnati, Ohio.
                                                                                   I
        Chemistry of the Soil. ACS Monograph No. 160, 2nd Ed.,                     m
        p. 264 (1964).Reinhold Pub. Co., N.Y.                                    •

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                                                                    36

        NITRATE NITROGEN IN BOTTOM SEDIMENTS - BRUCINE METHOD


1.  General Discussion
3.   Reagents
                                                                                    I
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                                                                                    I
     1.1  Principle:  See Standard Methods, 12th Ed., 1965, Method B,
          page 198-199.

     1.2  Interferences:  See Standard Methods, 12th Ed., 1965,                     I
          Method B, page 198-199.

2.   Apparatus                                                                      •

          Spectrophotometer for use at 1*10 mu, providing a light path
          of 1 inch or longer.
                                                                                    I



                                                                                    I
     3.1  Stock nitrate solution:  Dissolve 0.7218 g anhydrous potassium
          nitrate, KNO^, and dilute to 1,000 ml with distilled water.               •
          This solution contains 100 mg/1 N.                                        I

     3.2  Standard nitrate solution:  Dilute 100 ml stock nitrate solution          •
          to 1,000 ml with distilled water; 1.00 ml s 0.010 mg N.                    J

     3.3  Brucine-sulfanilic acid reagent:  Dissolve 1 g brucine sulfate            _
          and 0.1 g sulfanilic acid in approximately TO ml hot distilled            •
          water.  Add 3 ml concentrated HC1, cool, and make up to 100 ml.           "
          This solution is stable for several months.  The pink color
          that develops slowly does not affect its usefulness.                      •

     3A  Sulfurie acid solution:  Carefully add 500 ml of cone. H2SO.  to
          75 ml distilled water.  Cool to room temperature before use.              m
          Keep tightly stoppered to prevent absorption of atmospheric               •
          moisture.

k.   Procedure                                                                      •

     U.I  Transfer a portion of the previously clarified sample - 10 ml to
          one of the matched 1" colorimetric tubes.                                 •

     4.2  If less than a 10 ml aliquot is taken for analysis, add sufficient
          distilled water to make the volume 10 ml.                                 •

     k.3  Add 2.0 ml of 30$ NaCl to each tube and mix.                              ™

     k.k  Transfer the rack containing these tubes to an ice water                  I
          bath.                                                                     •
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                                                                    37


Nitrate Nitrogen in Bottom Sediments - Brucine Method



     k.5  Add 10 ml of the sulfuric acid solution to each tube (one
          tube at a time) mix and return to the ice bath.  Allow the
          tubes to reach thermal equilibrium before proceeding to
          the next step.


     k.6  Add 0.5 ml of brucine reagent (caution - poison) to each
          tube, mix thoroughly and return to the rack.


     U.7  Transfer the rack, with tubes, to a water bath and
          hold for 20 min. at a temperature just below boiling.


     U.8  At the end of 20 min. return the rack to the ice bath and
          cool to room temperature .


     4.9  Read the absorbance of the sample versus a blank in a
          spectronic-20, set at UlO n^ within 30 rain.


     4.10 Prepare a calibration curve using standards which were
          run concurrently with the samples .
5-   Calculations
     5.1  Wet Basis:
                                 OB Sample
                        _
                      OD Std     (grams sample

     5.2  Dry Basis:              in
             me/ke  -  rag As wet basis _
              6/^    % Solids (decimal fraction)


6.   References


     6.1  Standard Methods for the Examination of Water and Wastewater,
          12th ed., APHA, Inc., N.Y., 1965, 198-199-

     6.2  Chemical Analysis for Water Quality, A Training Course Manual,
          U.S. Dept. of Interior, FWPCA, Aug. 1966, pp 13, 11-13, 12.

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                                                                  38
     ORGANIC AND TOTAL KJELDAHL NITROGEN IN BOTTOM SEDIMENTS


    Organic nitrogen may be determined by digestion of the sample after
removal of free ammonia, with subsequent distillation and titration with          •
standard acid or as the difference between the value obtained for total           |
kjeldahl nitrogen and that for free ammonia.

                                                                                  I
1.  General Discussion
    1.1  Principle:  The kjeldahl method, using mercuric sulfate
                  as a catalyst, converts organically bound nitrogen in           I
         the trinegative state to ammonium bisulfate by digestion with            •
         sulfuric acid to which potassium sulfate has been added to raise
         the boiling point to 3^5° - 370°C.  The temperature should not           •
         exceed 3828C or loss of nitrogen will result.  After dilution,           |
         the solution is made alkaline with sodium hydroxide and the
         ammonia is distilled into 0.2N Ho SOj, or boric acid solution.            m
         If distillate is collected in 0.02N  H2SO]| acid the ammonia              •
         nitrogen should be determined by nesslerization and if distillate        ™
         is collected in 2% boric acid, the ammonia nitrogen should be
         determined by titration with 0.02N HgSO^,  using a mixed indicator.       •

    1.2  Interference:  In the presence of large quantities of nitrogen-
         free organic matter, it is necessary to add an additional 50 ml          •
         of sulfuric acid-mercuric sulfate-potassium sulfate solution for         •
         each gram of solid material in the sample.

    1.3  Storage:  Because organic nitrogen in unsterilized sewage and            •
         sludges is continually ammonified, the determination must be made        •
         on a freshly collected sample.  If the analysis cannot be made at
         once, the sample must be preserved with sufficient sulfuric acid         •
         to obtain a concentration of 1,500 mg/1 H2SOif or mo re (.8 ml/l).          |

2.  Apparatus                                                                     •

    2.1  Digestion apparatus, provided with a suction take-off to remove
         water vapor and sulfur trioxide fumes.

    2.2  Distillation apparatus:  See Nitrogen (Ammonia), Standard Methods        •
         for the Examination of Water and Wastewater. p. 187-193.
3.  Reagents

    3.1  Ammonia-free water
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                                                                  39
 Organic and Total Kjeldahl Nitrogen in Bottom Sediments



     3.2  Sodium Sulfate  (low in nitrogen).

     3.3  Mercuric Sulfate.

     3.k  Litnus Paper.


     3.5  Sodium hydroxide -sodium  thiosulfate solution:  Dissolve 500 g
          NaOH and 25 g Na2S203  •  5^0 in distilled water and dilute to
          1  liter.

     3.6  Phenolphthalein indicator solution:  Either the aqueous (a) or
          alcoholic  (b) solution may be used.

          a.  Dissolve 5  g phenolphthalein dlsodium salt in distilled
             water  and dilute to  1 liter.  If necessary, add 0.02N NaOH
             dropwise until a faint pink color appears.

          b.  Dissolve 5  g phenolphthalein in 500 ml 95 per cent ethyl
             alcohol or  isopropyl alcohol and add 500 ml distilled water.
             Then add 0.02N NaOH  dropwise until a faint pink color appears.

     3-T  Mixed  indicator:  Mix 2  volumes of 0.2 per cent methyl red in
          95  per cent alcohol with 1 volume of 0.2 per cent methylene
          blue in 95 per  cent ethyl alcohol.  This solution must be made
          fresh  every 30  days.

     3.8  Standard sulfuric acid titrant, 0.02N.  In this strength, 1.00 ml
          =   0.28 mg N.   Other strengths of standard acid may be used.

     3 . 9  Sulfuric acid,  cone .

     3.10 Dissolve 20 g anhydrous  boric acid, ^BO^, in ammonia-free
          water  and  dilute to 1 liter.

     3-11 Antifoam.


*.    Procedure

     k.l  Take either the residue  from the free ammonia analysis for
          organic nitrogen or a new sample for total kjeldahl nitrogen
          (1 or  2 g) .
         Add 35 ml of cone. HaSO^ plus 15 g Na2SOif, plus 3 g mercuric sulfate
         and a little antifoam and allow to boil until white fumes appear.

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                                                                 40                 I

Organic and Total Kjeldahl Nitrogen in Bottom Sediments                             •


    4.3  After fumes appear,  allow to digest for 30 minutes turning the
         flask from time to time.  Cool to room temperature.                        I

    4.4  To cooled sample,  add 500 ml  of  ammonia-free water, 0.3 ma
         (l dropper full) of  phenolphthalein or litmus paper, and then               •
         add 50 to 75 ml  of  50$ NaOH-thiosulfate  solution  being careful             |
         not to mix.  Place on distillation rack and mix.   If the solution
         is not red or litmus paper is not blue, add more NaOH until a               _
         red color appears  or litmus is blue.                                        I


    4.5  Distill over 300 ml  collecting the distillate in 50 ml  of                •
         2$ boric acid.  To this add 10 drops of mixed indicator and                I
         titrate with 0.02N H2S04 which has been standardized against
         0.02N Na2C03.

    4.6  Blanks and standards should be run in the same manner as the               •
         samples .

5«  Calculation                                                                     |

    5.1  Wet basis (titration)                                                      _

         mg/kg  *1 of 0.0? NH2S04 X 0.28
                    g   sample

    5.2  Dry basis                                                                   •

                   kg wet basis _                                           _
                  Solids (decimal fraction)                                          •

    5.3  Wet basis (coloriraetric)

         mg/kg » mg in Std x P.P. Sample x 1000                                      I
                 O.D. Std   (g   Sample
    5.4  Dry basis           in Aliquot)                                             |

         mg/kg B mg/kg wet baais
                                                                                     •
                                ^
                 56 solids  decimal fraction


6.   Nitrogen (Total  Kjeldahl) - (Bottom Sediments)

                                                                                     |
         Total kjeldahl nitrogen includes ammonia and organic nitrogen, but
     does not include nitrite and nitrate nitrogen.  The method is the same
     as  that for organic nitrogen, except that the ammonia removal step is            •
     omitted.  The procedure therefore begins with Sec. 4.  Place an appropriate       •
     aliquot of a well-mixed sample in an 800-ml kjeldahl flask. (l or 2 g)
                                                                                     I

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•               Organic and Total Kjeldahl Nitrogen in Bottom Sediments
                 7.     Reference
                       Standard Methods for the Examination of water and Wastewater.
                       12th ed.,  APHA,  Inc.,  N.Y.,  1965,  187-193.
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                                                                 te               I

                   OIL AND GREASE IN BOTTOM SEDIMENTS

1.  General Discussion                                                            •

    1.1  Definition:  Grease is defined as that material extracted by             •
         hexane from an acidified sample which would not be voltatilized          |
         during the procedure; it includes soaps, fats, waxes, and oils.

    1.2  Principle:  Drying of acidified sludges by heating leads to              •
         low results.  Magnesium sulfate monohydrate is capable of
         combining 75 per cent of its own weight in water in forming
         the heptahydrate.  Magnesium sulfate monohydrate can be used             I
         to dry sludge.  After drying, the grease can be extracted with           •
         hexane.

    1.3  Interference:  Elemental sulfur; certain organic dyes;                   •
         oxidation of extract and loss in weight of residue due to
         volatilization of low boiling components.                                •

    lA  Sampling and Storage:  Every possible precaution must be taken
         to obtain a representative sample.  When analyses can not be
         made immediately, samples may be preserved with 1 ml cone                •
         HgSOjj. for each 80 g of sample, or by freezing.                           •

2.  Apparatus                                                                     I

    2.1  Extraction apparatus, soxhlet or A.S".T.M. apparatus-.

3.  Reagents                                                                      •

    3.1  Hydrochloric acid cone.

    3.2  Magnesium sulfate monohydrate:  Prepare Mg SO^ • B^O by                  ™
         drying overnight a thin layer of Mg SO^ • T^O in an oven
         at 103°C.                                                                •

    3.3  M-Hexane, boiling point 69°C.

    3A  Grease free cotton:  Nonabsorbent cotton after extraction                •
         with N-hexane.
k.  Procedure
                                                                                  •
    U.I  In a 150 ml beaker weigh a 20 g sample of wet sludge, of
         which the dry-solids content is known.                                   •

    k.2  Acidify to a pH 2.0 (generally 0.3 ml cone HC1 is sufficient.
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 Oil and Grease in Bottom Sediments
    It.3  Add 25 g magnesium sulfate monohydrate.  Stir to a smooth
         paste and spread on the sides of the beaker to facilitate
         subsequent moisture removal.  Allow to stand until solidified
         15 to 30 minutes.

    k.k  Remove the solids and grind in a porcelain mortar.

    4.5  Add the powder to a paper extraction thimble.  Wipe the
         beaker and mortar with small pieces of filter paper moistened
         with hexane and add to the thimble.  Fill the thimble with
         small glass beads.

    k.6  Extract in a soxhlet apparatus using hexane at a rate of
         20 cycles per hour for 4 hours.

    k.J  If any turbidity or suspended matter is present in the
         extraction flask,remove by filtering through grease-free
         cotton into another weighed flask.  Rinse flask and cotton
         with hexane.


    k.8  Distill hexane from the extraction flask in water at 85°C.
         Dry by placing on a steam bath and drawing air through the
         .flask with a vacuum for 15 minutes.

    k.$  Cool in a desiccator for 30 minutes and weigh.

5.  Calculation

    5.1  Wet basis

              ,B mg of residue   x iQOO
                grams of sample

    5.2  Dry basis

           /    me/kg wet basis
         m&' kglS  $ solids (decimal fraction)

6.  Reference

    Standard Methods for the Examination of Water and Wastewater,
    12th Ed., AFHA,  Inc.,  N.Y.,  1965, '531-532.

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1.  General Discussion
    1.2  The extremely high dilutions used will help to minimize any
         interferences due to toxic materials.

2.  Apparatus
                                                                                   I
     IMMEDIATE DISSOLVED OXYGEN DEMAND AND BIOCHEMICAL OXYGEN DEMAND                 _
                             OF BOTTOM SEDIMENTS                                     •
I
    1.1  See Standard Methods for the Examination of Water  and Waste-
         Water,  12th Edition, page *H5.•
I
    2.1  Incubation bottles, 300 ml bottles with water seal.  Ground-              •
         glass stoppered bottles should be cleaned with a good                     •
         detergent or dichromate cleaning solution and thoroughly
         rinsed v-i-i-.h distilled HoO and drained.  Satisfactory water                •
         seals are obtained by adding water to the top of the BOD                  |
         bottles each morning.

    2.2  Air incubator, thermostatically controlled at 20°C ± 18C                  I
         (All light should be excluded to prevent formation of DO
         by algae in the sample.)

3-  Reagents                                                                       •

    3.1  Distilled water used for solution and for preparation of the              •
         dilution water must be of the highest quality, distilled from             |
         a block tin or all-glass still, contain less than 0.01 mg/1
         copper, and be free of chlorine, chloramines, caustic                     •
         alkalinity, organic material, or acids and have a specific                •
         resistance of 750,000 OHMS or higher.  The distilled water
         used for dilution water should be aged or stabilized in the
         dark for 30 days prior to using it.                                       •

    3.2  Phosphate buffer solution:  Dissolve 8.5 g potassium
         dihydrogen phosphate, KH^^k > 21-T5 g dipotassium hydrogen                •
         phosphate, ^HPOj^ ; 33.4 g disodium hydrogen phosphate hepta-              |
         hydrate, Na2HPO^ • 71^0 and 1.7 g ammonium chloride, NH^Cl,
         in about 500 ml distilled water and dilute to 1 liter.                    .
         The pH of this buffer should be 7.2.                                      •

    3.3  Magnesium sulfate solution:  Dissolve 22.5 g MgSOj,.  • 7H20
         in distilled water and dilute to 1 liter.                                 •

    3A  Calcium chloride solution:  Dissolve 27.5 g CaClg in distilled
         water and dilute to 1 liter.                                              •

    3.5  Ferric chloride solution:  Dissolve 0.25 g of FeCl^  . 6H20 in
         distilled water and dilute to 1 liter.                                    «




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Immediate Dissolved Oxygen Demand and Biochemical Oxygen remand of
Bottom Sediments


     3.6  BOD standard:  Dissolve 0«l50 g each of reagent grade glucose
          and glutamic acid which have been dried at 103°C for 1 hour
          in distilled water and dilute to 1 liter.

     3.7  Seeding material:  A satisfactory seed may be obtained by-
          using the supernatant liquor from raw domestic sewage which
          has been stored at 20°C for 2h hours.

U.   Procedure

     liol  Preparation of dilution water:  The distilled water used should
          have been aged in cotton-plugged bottles for a sufficient length
          of time to insure stabilisation and to give blank depletions of
          not more than 0.2 mg, preferably 0.1 mg, oxygen.  Usually 30
          days are required.  The water should be aerated just prior to
          use to insure saturation at 20°C.              Add 1 ml each
          of phosphate buffer, magnesium sulfate, calcium chloride, and
          ferric chloride solutions for each liter of dilution water.
     k.2  Seeding:  Add proper amount of seed (to give 2 mg/1
          depletion), which has been filtered through
          two thicknesses of Kimwipes for each liter of water.  The
          amount of seed required may vary with the source of seed and
          will have to be established through experience0

     lj.3  Weigh appropriate size sample directly into the BOD bottle*
          suggested weight 6 grams.

     luU  Fill BOD bottle with dilution water and place in incubator.

     iu5>  Make sure that there is a water seal in the neck of each
          bottle when placed in the incubator.  Each morning replenish
          the water seal on all bottles.

     Iu6  Determination of initial DO:  Determine the initial DO of
          the dilution water by means of the azide modification of the
          iodometric method0  This DO value is used as the initial DO
          of the sample.  This method is used because it is not possible
          to obtain an exact initial DO of the bottom sediment due to
          the rapid consumption of oxygen.
          (a)lf a DO probe is used, IDOD and BOD can be run on the same bottle.
     lu?  Incubation:  Incubate the blank (dilution water) and the diluted
          samples for £ days at 20°C.  Then determine the DO in the
          incubated samples using the above methods.  Those dilutions
          showing a residual DO of 2 mg/1 and a depletion of 2 mg/1 are
          considered the most reliable.

     Ijo8  Dilution water control:  Fill two BOD bottles with unseeded
          dilution water.  Stopper and water-seal one of these for
          incubation.  The other bottle is for determining DO prior to
          incubation.  The DO results on these two bottles are used as
          a       check on the quality of the unseeded dilution water.
          The depletion obtained should not be more than 0.2 ml and
          preferably not more than Oel ml.

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 Immediate  Dissolved Oxygen Demand  and  Biochemical  Oxygen Demand  of       ^6          •
 Bottom Sediments
      12th ed., APHA,  Inc.,  N.Y.,  1965,
                                                                                     I
      Uc9  Glucose-glutamic  acid check:   To  check the  dilution water,
           the seed material and the technique  of the  analyst, make up               •
           a two  percent dilution of the  primary BOD standard.  Fill                  J
           three  bottles and incubate at  20°C for 5>  days.  A  two  percent
           dilution of the BOD standard solution should  give  a BOD of                 _
           218 mg/1 with a standard deviation of +_ 11  mg/le   If the                   •
           results  deviate from this value appreciably,  the quality of               •
           the seed, water and the technique is questionablec

 5>.    Immediate Dissolved Oxygen Demand                                               •

      5,1  This test is carried out in the same manner as  the 5-day                   •
           BOD test, except that the sample  is  held  for  only  l£ minutes               J
           and the  DO is determined.

      5.2  Calculation:                                                              I
           Wet basis      /_.    n \                                                  •
           I DOD  mgAg c  ^  0 "  3-  XO-3 ___   x 1000
                          grams  of sample in aliquot                                 •
           Dry basis                                                                 *
           I DOD  mg/kg * mg/kg I POP  (wet basis)                                    _
 6.    BOD Calculation    * solids (decimal  fraction)                                 |

      6.1  Wet basis                                                                 _

                       *                                  x 1000                      I
                    grams of sample in aliquot
      6,2  Cry basis                                                                 •

           mgAg " mg/kg (wet basis) _
                   $ solids" (decimal fraction)                                        I

      6e 3  Definitions

           DQ * DO of  original  dilution water.                                        |

           D^ « DO of  diluted sample  15 min0 after preparation.                       _

           Dp « DO of  diluted sample  after incubation (after 5 days),                 ™

7-    Reference                                                                      •

      Standard Methods for the  Examination of Water and Wastewater.
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                                                                  vr

            OXYGEN UPTAKE OF BOTTOM SEDIMENTS
General Discussion


The oxygen demand of bottom sediments is a complex phenomenon
occurring at the water-sediment interface, where chemical and
biochemical oxidation and reduction reactions take place at
various rates depending upon conditions existing at a particular
time.

Since bottom sediments tend to be of a reducing character, water
flowing over these sediments lose oxygen to the sediment inter-
face, which becomes an oxidized microzone of ferric oxide.  Under
these conditions, the water-sediment interface is a brown color
and the radox potential in this system is greater than 0.2 volts.
When stagnation progresses and conditions become anaerobic, the
radox potential is less than 0.2 volts and the brown color dis-
appears .


The presence or absence of the oxidized microzone makes an
immense difference in the amount and nature of materials that are
leached from these sediments and the rate of oxygen uptake from
the waters above.


The amount of oxygen consumed by bottom sediments is often great
enough so that it causes the overlying waters to become void of
oxygen several feet above the water-sediment interface.  There-
fore, it is important to know the characteristics of these bottom
sediments and their oxygen uptake rates.


Apparatus


2.1  Laboratory Oxygen Analyzer.

2.2  Magnetic Stirrer with one-inch teflon coated magnets.

2.3  Incubator,  20°C.


2.k  Wide-mouth Cylindrical Jars with screw cap and sealed probe;
     minimum mouth opening of 11 cm; height 25 cm;  for use in
     making D£ uptake apparatus (see attached diagram).

2.5  Glass Petri Dish with cover.


2.6  Glass Petri Dish Support.


2.7  Asbestos Sheet,  15 x 15 x 0.5 cm.

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Figure No. 1
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Oxygen Uptake of Bottom Sediments

3.  Reagents

    3.1  Distilled water:  Water used for solutions and for preparation
         of the dilution water must be of the highest quality,  distilled
         from a block tin or all-glass still, contain less than 0.01 mg/1
         copper, and be free of chlorine, chloramines,  caustic  alka-
         linity, organic material, or acids.

    3.2  Phosphate buffer solution:  Dissolve 8.5 g potassium dihydrogen
         phosphate, KH^POjj; 21.75 g dipotassium hydrogen phosphate,
                 33.If g disodium hydrogen phosphate heptahydrate,
                '7H20, and 1.75 8 ammonium chloride, NHj^Cl, in  about
         500 ml distilled water and dilute to 1 liter.   The pH  of  this
         buffer should be 7.2 without further adjustment.  If dilution
         water is to be stored in the incubator, the phosphate  buffer
         should be added just prior to using  the dilution water.

    3.3  Magnesium sulfate solution:   Dissolve 22.5 g MgSO^-THgO in
         distilled water and dilute to 1  liter.

    3.1*.  Calcium chloride solution:   Dissolve 27.5  g anhydrous  CaCl2 in
         distilled water and dilute to 1  liter.

    3,5  Ferric  chloride solution:   Dissolve,  0.25 g FeCl3'6H20  in
         distilled water and dilute to 1  liter.

    3.6  Acid and alkali solutions,  IN.   For  neutralization of  waste
         samples which are either caustic  or acidic.

    3.7  Sodium  sulfite solution,  0.025N.   Dissolve 1.575 g anhydrous
         NagSO  in 1,000 ml distilled water.   This  solution is  not
         stable  and should be prepared daily.

    3.8  Seeding material:   A satisfactory  seed may be obtained by
         using the supernatant liquor from  raw domestic sewage  which
         has  been  stored at 20°C for  2k hours.

    Procedure

    k.I   Preparation  of dilution water:   The  distilled water used
         should  have  been stored in cotton-plugged  bottles  for  a
         sufficient length of time to become  saturated with DO.  The
         water may also be aerated by shaking  a partially filled
         bottle  or with a. supply of clean compressed air.   Situations
         may  be  encountered where it  is desired to  use stabilized
         river water  to check stream  performance with laboratory
         procedure.   The  distilled water used  should be as  near 20°C
         as possible  and of the  highest purity.  Place the  desired
         volume  of distilled water in a suitable bottle and add 1 ml
         each of phosphate  buffer, magnesium  sulfate, calcium chloride,
         and  ferric chloride solutions for  each  liter of water.

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                                                                                    I
Oxygen Uptake of Bottom Sediments
                                                                                    I
    k.2  Place a covered petri dish containing a weighed amount of
         well-mixed sediment in p
         oxygen uptake apparatus.
     well-mixed sediment in place, as shown in the sketch of                    I
     Fill the jar with dilution water, measuring volume  of
     added water, then remove the petri dish cover.

     Insert oxygen probe with cap after standardizing probe
         as shown below:
                                                                                    I
         jam ex o y-xjgeu pjuue WJLUII cap «u i/cz Bi/anucLrui^ing piuoe                      •
         against Winkler DO method, following the manufacturer's                     I
         manual and place sample in a 20°C incubator.

    4.5  Seal system and with magnetic stirrer set to give approxi-                  I
         mately the same flow occurring in the lake or streams.  Take                •
         dissolved oxygen readings at various time intervals.  In
         general, time intervals of 0, 0.5, 1, 2, 3, k, 5, 6, 8, 10,                 •
         15, 20 and 30 days are satisfactory.  The frequency of read-                |
         ings is dependent upon the rate of oxygen uptake.

5-  Calculation of oxygen uptake                                                     •

    Wet basis                           Dry basis

    A x 1000  ** mg/kg                    wgAg  B rcgAg wet  basis                       •
    B                                            % solids (decimal fraction;

    A « Initial reading - Final reading                                             |

    B * Weight of  sample in grams per liter                                          _

    Oxygen uptake may be calculated on the basis  of weight or unit area              ™
    by substituting surface area of sediment exposed to the dilution
    water in place of B (the weight of sediment)  in the above formula.               •

6.  Calculation of K^ values
6.1  From the 5-d.ay BOD values (y), calculate the daily differences             •


                                                                                I
 t              Op-demand           Differences



0.5                                    0.72                                  I
 1                0.72                                                       •
 1.5                                   0.48
 2                1.20                                                       •
 2.5                                   0.35                                  •
 3                1-55
 3-5                                   0.27
 U                1.82
 U.5                                   0.20
 5                2.02
                                                                                     •
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                                                                                   51

            Oxygen Uptake of Bottom Sediments
                6.2  Plot the differences on semi-log paper with differences
                     on the log scale and time on linear scale.   The time
                     coordinates are 0-5, 1«5> 2.5,  etc.

                6.3  Draw a straight line through the points,  extending the
                     line to t = 0 (left margin).

                6.4  Determine the slope of this  line by using values at 5 days
                     and 0 days.

                     5  days = 0.175 and 0 days =  0.74.
     Slope X 100 = % remaining in 5 days = 23.7$


6.5  Using the formula, log % BOD remaining = 2.0-K.j^t; K^ can be
     calculated as follows:


          log     23.7   r2.0  -1C 5
                   1.374 r2.0  — KJ5
                   K,     =2.0  -1.374
                               5

                   K.J,     =0.13


6.6  The formula Y r L  (l-10~ 1 ) and the above data will permit
     the calculation of.L,  L being the ultimate BOD, Y the 5~day
     BOD, and (l-10"%^Hhe fraction oxidized.


     Fractions oxidized =  1—fraction remaining


                        =  1-0.237


                        =  0.763

     Therefore 2.02 :   L X 0.763
•                                L =   2.02
                                       0.763
                                  L -   2.7 mg/1

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                                                                      52

Oxygen Uptake of Bottom Sediments
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    References

    7-1  Sheehy, J. P.  "Rapid Methods for Solving Monomolecular                  •
         Equations," J.W.P.C.F., 32:646, June 1960.

    7-2  Moments Methods:  Moore, E.W., Thomas, H.A., and Snow, W.B.,             |
         "Simplified Method for Analysis of BOD Data,"  Sewage and
         Industrial Wastes, 22; 10, 1950.                                         .

    7-3  Tsivoglou, E.G., "Oxygen Relationships in Streams,"  Technical
         Report W-58-2, Robert A. Taft Sanitary Engineering Center, 1958.

    7.4  Muhlman, F.W., "Oxygen Demand of Sludge Deposits,"                       I
         J. Sewage Works, 10, 613 (1938).

    7.5  Rudolfs, Willed,  "Relation between Biochemical Oxygen                   |
         Demand and Volatile Solids of Sludge Deposits in the
         Connecticut River.  J. Sewage Works, U, 317 (1932).                      _

    7.6  Beckman Instructions #1223, Beckman Instruments, Inc. (1962).


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                                                                     53
                         CHLORINE DEMAND OF BOTTOM SEDIMENTS

1.  General Discussion

   "The chlorine demand of a water is caused Toy such inorganic reductants
    as ferrous, manganous, nitrite, sulfide, and sulfite ions.  Ammonia
    and cyanide consume considerable chlorine during the free residual
    chlorination process.  Chlorine substitutes on phenols and other
    similar aromatic compounds to form chloro derivative compounds, but
    may also oxidize the aromatic compounds when larger amounts of chlorine
    are added.  It may also react with ammonia and naturally occurring
    amino compounds to form chloramines with an active or oxidizing chlorine
    atom.  The destruction of the chloramine compounds can be achieved by
    the addition of more chlorine and subsequently with the addition of
    enough chlorine a free available residual (hypochlorous acid) may be
    attained.

   "The chlorine demand of water is the difference between the amount of
    chlorine applied to a treated supply and the amount of free, combined,
    or total available chlorine remaining at the end of the contact period.
    The chlorine demand of any given water varies with the amount of chlorine
    applied, time of contact, pH, and temperature.  For comparative purposes
    it is imperative that all test conditions be stated.  The smallest amount
    of residual chlorine considered significant is 0.1 mg/1 Cl.  Presented
    here is  a method for laboratory use and a field procedure which gives
    less exact results.

     1.1  Principle:


          The laboratory method  is designed to determine the  so-called
          immediate demand as well as other demands at longer contact
          periods.  Chlorine demand determinations are made to determine
          the amount of chlorine that must be applied to a water to pro-
          duce a specific free,  combined, or total available chlorine
          residual after a selected period of contact.  If the amount of
          chlorine applied to waters containing ammonium or organic
          nitrogen compounds is not sufficient to reach what is termed
          the "breakpoint" chloramines and certain other chloro derivatives
          which react as combined available residual chlorine are produced.
          When sufficient chlorine has been added to reach the breakpoint,
          which depends on pH, ratio of chlorine to nitrogenous compounds
          present, and other factors, subsequent additions of chlorine
          remain in the free available state." (Standard Methods, 12th
          Edition, pp.112-113)

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                                                                                   I
Chlorine Demand of Bottom Sediments                                                •
2.   Reagents
     2.1  Acetic acid, cone.                                                       I
     2.2  Potassium iodide, crystals.
     2.3  Standard thiosulfate,  0.037 N.

     2.4  Solution of starch, 5 grams of starch per liter of water.
          detecting the end point in the presence of turbidity pro-
          duced by the sediment.
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     2.5  Chlorine solution adjusted so that 1 ml contains approximately
          1 mg of chlorine.                                                        •

     2.6  Aged distilled chlorine free water.

3.   Apparatus                                                                     •

     3.1  Stirrer and magnetic stirring bars.

     3.2  500 ml erlenmeyer flasks-                                                |

     3-3  50 ml buret.                                                             _

     3-4  Aluminum foil.                                                           •

4.   Procedure                                                                     I

     4.1  Weigh 1-2 grams of well-mixed bottom sediment onto prepared
          aluminum strips.                                                         •

     4.2  Wash sediment from aluminum strips into «nerlenmeyer flask
          containing 200 ml  of aged chlorine free water by moving
          aluminum strip up and down below surface in flask.                       I

     I}.. 3  Pipet 20 ml  of standardized chlorine solution into sample
          with rapid stirring to insure instantaneous mixing and then              •
          stir slowly in darkness for exactly 15 minutes.                          |

     4.14.  Add 5 ml  of acetic acid, approximately 1 gram of potassium              _
          iodide, 1 to 2 ml  of starch solution and titrate with                   •
         0.0370 N thiosulfate to the point of disappearance of the                 ™
          blue color.  Note - it is necessary to add the starch at
          the beginning of the titration due to the difficulty in                  •
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 I
 —             Chlorine Demand of Bottom Sediments

                5-   Calculation
 I                  One ml of 0.037 N thiosulfate is equivalent to 1.3010 mg chlorine.
                     15-1 Wet Basis:
                            /.   _,  ,    ,    mg Cl added — mg Cl residual  ., ,„.-.
                         ing/kg Cl demand =  "B ^ Qf sedim^t -  X 1000
 I                  5.2 Dry Basis:
                            /     mg/kg vet basis
                         mg/Kg *
                                  ^ s0ids (decimal fraction)
 |              6.   Reference
                     •Standard Methods  for the Examination of Water and Wastewater,
                     12th  ed., APHA, Inc.,  N.Y.,  1965,  112-113.

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                                                                       56
                    PESTICIDES IN BOTTOM SEDIMENTS
1.   General Discussion
     2.4  Thin layer chromatography equipment.

     2.5  Gas chromatograph equipped with microcoulometric titration
          cells.
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                      Preparation of Sample
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     1.1  Principle:  Pesticides sorbed on particulate matter may be
          desorbed by the continuous Soxhlet Extraction of a ground and             •
          dried bottom sediment sample with suitable organic solvent(s).            |
          The extract containing the pesticide(s) is cleaned up and
          analyzed by gas chromatography, thin layer chromatography, and            _
          infrared spectrescopy.                                                    I

2.   Apparatus

     2.1  Soxhlet extractor, flask capacity (500 ml), with Allihn                   I
          condenser.

     2.2  Extraction thimbles, Whatman single thickness, 43 x 123 mm or             •
          a convenient size.

     2.3  Chromatographic tube 20 x 400 mm.                                         I
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3.   Reagents

     3.1  Hexane, redistilled in glass b.p. 68° - 69°C.

     3.2  Ethyl ether, redistilled in glass b.p. 34° - 35 °C.                        •

     3.3  Chloroform, redistilled in glass b.p. 60° - 6l°C.                         •

     3.4  6 parts ethyl ether and 94 parts hexane.

     3-5  15 parts ethyl ether and 85 parts hexane.                                 •

     3.6  30 parts ethyl ether and 70 parts hexane.

     3.7  50 parts ethyl ether and 50 parts hexane.                                 |

     3.8  Florisil 60-100 mesh commercial products of this grade may
          be reactivated by heating 5 hours at 130°C.                               •
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                                                                        57


Pesticides in Bottom Sediments,  Preparation of Sample



k.   Procedure


     k.l  Preparation for Extraction


          a.   Allow the bottom sediment sample to dry thoroughly at
               room temperature for about a week or cover samples with
               open mesh cheesecloth and accelerate moisture removal
               employing a cooling fan.   This is best accomplished by
               breaking up the large particles and spreading out in a
               glass dish or aluminum foil.


          b.   Grind up sample with a mortar and pestle and pass it
               through a 30 mesh sieve.


          c.   Select a 100 g   aliquot  and place in a pre-extracted
               thimble.  Place a small wad of pre-extracted glass wool
               or cotton on top of the sample.


     k.2  Soxhlet Extraction


          a.   Using 300 ml chloroform,  extract bottom sediment sample
               for 18 hours (at 60°C).


          b.   Transfer extract to 250 ml beaker and concentrate on
               a steam bath to about 10  ml; add one drop 0.005$ paraffin
               oil in hexane prior to evaporation.


     ^.3  Wet Extraction Procedure


          Extraction of soils and bottom sediments.  Weigh a lOOg sample
          into a liter erlenmeyer flask.  Add distilled water to effect
          a slurry.  Add 2 ml of extraction solvent (hexane/IPA,  3:l)
          per gram of sample and shake vigorously for 20 minutes using a
          wrist action shaker or equivalent.  Decant and collect the
          hexane phase into a separatory funnel.  Repeat extraction of
          the bottom sediment/aqueous phase two more times quantitatively
          decanting the hexane portions  each time into the separatory
          funnel.  Wash any remaining alcohol from combined hexane extracts
          with water,  dry over sodium sulfate,  and concentrate to an approp-
          riate volume.

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Pesticides in Bottom Sediments, Preparation of Sample
     k.k  Column Cforomatography


          a.   Prepare a five inch column with 3A" at top and 1/2" at
               bottom anhydrous sodium sulfate plus 15 gram Florisil
               which has been activated 5 hours at 130°C and stored in
               a desiccator to cool; or for samples containing large
               quantities of waxes and pigments, use a column containing
               a mixture of Nuchar C-190 (5g) plus Celite (lO grams).
               Column may be reduced to 3/8" diameter x 12" (made from
               glass tubing drawn to approximately l/l6"tip) packed with
               1/2" x 1/2" glass wool; reagents must be reduced by a
               factor of 10.
          b.

          c.




          d.




          e.
 Pre-wet the column with 50 ml of hexane.

 Place the extract on the column and elute into a 200 ml
 volumetric flask with 200 ml 6%, 15% and 30$ or $0% ethyl
 ether in hexane.

 Concentrate the sample from step c to approximately 5 nJ-
 in a 250 ml beaker; then transfer to a 15 ml centrifuge
 tube and concentrate to a volume of 0.1 to 1 ml.

 The following pesticides are eluted by the above mixtures
 of ethyl ether and hexane.
Lindane
BHC
Kelthane
Aldrin
Heptachlor
DDE
TDE (DDD)
DDT
Perthane
Heptachlor or
Methoxychlor
Toxaphene
Strobane
Chlordane
Endosulfan I
             ETHYL ETHER IN HEXAHE


               15*

               Dieldrin
               Endrin
               Endosulfan II
               Lindane
               Kelthane(pos s ible
               trace of total)
30% or 50%

Generally elute
Thiophosphate Pesticides

50$ E. Ether elutes
Guthion
epoxide
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                                                                      59
                         PESTICIDES IN BOTTOM SEDIMENTS

                   Gas Chromatography with Electron Capture

1.   Nonpolar Column

     1.1  General Discussion:   This procedure is  applicable to the
          analysis of most chlorinated hydrocarbons,  a number of esters
          of the phenoxy-alkanoic acid herbicides,  and a few organic
          phosphates.

     1.2  Apparatus:

          a.   Gas chromatograph equipped with dual glass columns  and
               electron capture detector,  spare electron capture detector
               recommended.

          b.   Strip chart recorder — 1 mv.

          c.   Microsyringes—10- and 50 ul capacity.


          d.   Glass Columns:   Non-Polar  6 feet by  1/4"  O.D.  packed
               with Gas Chrom Q 60/80 mesh, coated  with  5$ by weight
               of OV-17.

          e.   Nitrogen tank equipped with regulator valve and tubing
               to connect with gas chromatograph.
     1.3  Reagents:

          a.
     Stock pesticide standards are made to contain 1 ng/ml
     of lindane; heptachlor; aldrin;  dieldrin;  endrin;  o,p-DDT;
     p,p'-DDT; DDD, DDE, methachlor,  and chlordane in hexane.

b.   Pesticide working standards - Pesticide stock standards
     are diluted with hexane to give the following concentrations:
               Standard 1

               Lindane
               Heptachlor
               Aldrin
               DDE
               o, p-DDT
               p, p'-DDT
                    ng/nl
                    0.5
                    1.0
                    1.0
                    7.0
                    8.0
                    8.0
Standard 2

Dieldrin
Endrin
ngAa
3.0
8.0
                                                    Standard 3
Chlordane     10.0
Methoxychlor  10.0

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                                                                        60


Pesticides in Bottom Sediments,  Gas Chromatography with Electron Capture            •


     l.k   Column Conditioning:

           a.   Install the packed column in the gas chromatograph                  •
                without connecting to the detector device.   Heat the
                column to 270°C without gas flow for two hours.   Allow              •
                the column to cool to 225°C and maintain this temperature           |
                for 10 minutes;  then adjust carrier gas to  about 50 ml per
                minute.  After one hour,  increase column temperature to             •
                2ifO°C and condition the column for 2k-k8 hours.   Cool the           I
                column down to operating temperatures l80-215°C  and hook up
                to the detector system.  Columns prepared and conditioned
                in the prescribed manner should yield good  chromatograms            I
                without major losses of heptachlor or DDT with no further           •
                treatment.  Even with the most advanced equipment,the
                analyst is reminded that the system can be  no better than           •
                the condition of the column.  Injection of  samples which            jj
                have not been adequately prepared will undoubtedly shorten
                the life of the column and yield poor quality chromatograms.         _

           b.   Conditioning of the column can also be done in a separate           *
                oven capable of maintaining temperatures up to 270°C and
                equipped to maintain a flow of carrier gas  through the              •
                column.  This permits conditioning of a standby  column              |
                while the instrument is being used for residue analysis.

           c.   At the end of the preliminary conditioning  period,  the              I
                chromatograpnic system should be evaluated  using pesticide
                mixtures and standards injected singly.  Recovery of
                chlorinated hydrocarbons should be 75$ or higher, and               I
                peak shapes should be symmetrical.  Evaluation of the               •
                chromatographic column can also be done by  using the
                pesticide endrin as a guide.  It has been shown  that columns         •
                not properly conditioned give two peaks for this pesticide          |
                at low temperatures owing to catalytic decomposition,  but,
                upon further conditioning,  it will give only one peak.              «
                Double peaks, however, still can be obtained at  high temp-          •
                eratures as a result of thermal decomposition.  Individual
                laboratories must judge whether or not the  chromatographic
                column is operating at maximum efficiency.   Before the              I
                column is ready for residue analysis,  it should  give high           I
                recoveries of most pesticide standards, good resolution,
                and symmetrical peaks.                                              •
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Pesticides in Bottom Sediments, Gas Chromatography with Electron Capture
           d.   If further conditioning of the column is indicated,  the
                column should be heat-conditioned for an additional twenty-
                four hours before reevaluation.  It may also be necessary
                to make several injections of 25 ul each of the strong
                pesticide mixture.  This should be followed up with several
                injections of 5 ul each of the moderate pesticide mixture;
                if this still does not give a satisfactory chromatogram,
                injection of purified sample extracts may be necessary.
                If these materials are available, it is recommended that
                several 10 ul injections be made in an attempt to condition
                the column for residue analysis.

           e.   Once a column has been properly conditioned and is ready
                for residue analysis, it may be necessary to make several
                injections of the standard pesticide mixture each morning
                prior to running sample extracts.

     1.5   Procedure:

           a.   After the nonpolar column has been thoroughly conditioned
                as described in Section 1.4, on conditioning of the column,
                it is ready for use.  The operating parameters for this
                column are as follows:  column temperature, 195°C; injection
                block, 230°C; electron capture detector, 200°C; the flow  rate
                of nitrogen through the column is maintained at 120 ml/min.,
                or other optimum conditions.

           b.   Optimum operating conditions for the electron capture
                detector must be determined in the individual laboratory  by
                varying the voltage applied to the detector to find a region
                where sensitivity is good and response is linear.

           c.   For the operation of the gas chromatograph, follow instructions
                in the manufacturer's manual.

           d.   To obtain relative retention times on this column using the
                electron capture detector, individual pesticides, mixtures
               (containing aldrin), and unknowns should be injected at the
                1  to 10 ng level.

           e.   Relative Retention Time (R.R.T.) s
                Pesticide's Actual Retention Time in Minutes
                Actual Retention Time of Aldrin in Minutes
                (Sulphenone is usually the referenced pesticide when computing
                 R.R.T. of thiophosphate pesticides.)

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                                                                                       I

Pesticides in Bottom Sediments, Gas Chromatography with Electron  Capture              •


2.   On the Polar Column                                                               _

     2.1   General Discussion: This procedure is applicable to  the                     ™
           analysis of chlorinated hydrocarbons, a few organic  phosphates
           and some of the phenoxyalkanoic acids and their esters.   This              •
           column has been selected to give a different pattern of  separation         |
           of the pesticide standards in order to facilitate  identification.

     2.2   Apparatus:                                                                  •

           a.  (Same as for nonpolar column except the column)

           b.   Glass columns - 6 feet in length by 1/4" O.D. packed with              •
                Gas Chrom Q 60/80 mesh coated with 5$ QF-1 plus 3%  DC-200.

     2.3   Reagents: (Same as given for non-polar column)                              |

     2.4   Column Conditioning:  (Same as for nonpolar column)                         _

     2.5   Proc edure:

           a.   The operating conditions for the polar column are as                  I
                follows:  column temperature, l80°C; injection  block, 230°C;           I
                detector, 200°C; flow rate of nitrogen at 100 ml/min.  The
                optimum operating voltage for the direct current  mode of              •
                operation of the electron capture detector should be deter-           •
                mined by injecting a standard solution of lindane and adjust-
                ingthe voltage until maximum sensitivity and  a  linear res-             _
                ponse is obtained.  If the two are not compatible,  a voltage          •
                •i c r»Vi/~i<2*ar» -hViQ-h wf 1 1 irl f*~\ H n ~lTYi«=»nT» Y»£ic:Tw~in <=:#=» n n  "hVifa  vssncr^ of*           *B
                is chosen that will yield  a linear response in the range of
                1-10 ng of  standards.   Then make unknown sample injections
                in the same manner as  standards.
3.   Calculations

     Snploy the  calculations  given under Microcoulometric Section
     "Measurement  of Unknown,"
I
                The polar  column  should be used to assist in identification
                of unknown compounds  chromatographed on the nonpolar column.          «
                This type  of  confirmation  is  of great importance when using           I
                the electron  capture  method since some non-pesticide impurities
                in sample  extracts  give high  responses with this detector.
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                                                                      63


Pesticides in Bottom Sediments, Gas Chromatography with Electron Capture



4.   References

     4.1   Burchfield, H.P. and Johnson, D.E., 1965.  Manual of Methods
           for the Analysis of Pesticide Residues.  Southwest Research
           Institute  (Prepared for DHEW, PHS), Vol. I and II.

     4.2   Organic Pesticide Subcommittee of Committee on Methods
           Validation and Analytical Quality Control, FWPCA, 1968.
           Provisional Interim Official Methods for Chlorinated Hydro-
           carbon Pesticides in Water and Wastewater by Gas Chromatography,
           1968.

     4.3   Breidenbach,  A.W.,  et. al.  1966.  The Identification and
           Measurement of Chlorinated Hydrocarbon Pesticides in Surface
           Waters.  U.S. Department of Health, Education,  and Welfare,
           Public Health Service WP-22 November.

     4.4   Burke, J. and Giuffrida, A.L.  1964.  Investigation of Electron
           Capture Gas Chromatography for the Analysis of Multiple
           Chlorinated Pesticide Residues in Vegetables.  J. Assoc.
           Off. Agr. Chemists  47:326.

    4.5    Hundley,  J. C.  et. al.,  1964.  Pesticide Analytical Manual
           (Vol.  II).  U.  S. Department  of Health, Education, and
           Welfare,  Food and Drug Administration.


     4.6   Shuman, H. and Collie, J.R.,  1963.  Gas Chromatographic Columns
           for Optimum Recovery of Chlorinated Pesticides.  J. Assoc. Off.
           Agr. Chemists. 46:  992.


     4.7   U.S. Dept. of the Interior, FWPCA, 1968.  Report on Insecticides
           in Lake Michigan prepared by Pesticide Committee of the Lake
           Michigan Enforcement Conference,  Donald I. Mount, Ph. D.,
           Chairman, National  Water Quality Laboratory,  Duluth,  Minn.

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1.   General Discussion
     3.2   Stationary phase DC-200,  QF-1,  0V IT,  OV-101,  etc,

     Procedure
           ^Recommended condition may be adjusted depending upon
            choice of column.
                                                                                      I
                       PESTICIDES IN BOTTOM SEDIMENTS                                 •

                      Microcoulometric Gas Chromatograph
                                                                                      I
     Principle:  The microcoulometric titration cell used as a gas                    •
     chromatographic detector has for its basis the coulometric                       ™
     titration of resultant ions with electrically generated ions.
     Inorganic ions are produced when the effluent from the gas                       •
     chromatographic column enters the combustion furnace where any                   I
     organic pesticide hydrocarbons present are broken down into
     inorganic species.                                                               •

2.   Apparatus
     2.1   Dohrmann Model C-200 microcoulometer equipped with microcoulometric        •
           titration cells.                                                           •


                                                                                      I
           titration cells.

     2.2   Glass,  aluminum,  or stainless steel 6 ft.  column.

3.   Reagents

     3.1   Solid support Chromosorb W,  P, G or Gas Chrom Q,  or                       •
           Anachrom ABS (acid washed dimethyl-dichlorosilance treated.
           AW-DMCS).
                                                                                     I
           Recommended Operating Conditions*

           a.   Furnace temperature 850°C.                                            •

           b.   Inlet temperature 300 °C .

           c.   Og flow rate 50 cc/min.                                               |

           d.   N2 sweep rate 20 cc/min.                                               M

           e.   Column temperature 190 °C or other temperature.

           f .   Plow rate (Na) 100-200 ml/min.                                        •

           g.   Diameter 1/8 inches (inside diameter) .
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                                                                     65
Pesticides in Bottom Sediments,  Microcoulometric Gas Chromatograph
     4.2   Measurement of Standard Chloride Solution

           a.   Set FUNCTION switch to BIAS READ and adjust BIAS
                MILLIVOLTS with BIAS SET knob to 12 divisions to the
                left of zero.  This supplies 2kO bias millivolts to
                the cell.

           b.   Adjust stirring rate in titration cell to about 1/2
                full position (NOTE:  Be sure the magnet does not
                decouple.)

           c.   Set FUNCTION switch to GENERATOR READ.  Generator
                volt meter should read 0 or to the left of 0.  If the
                meter reads to the right of 0, add fresh electrolyte
                to the cell and flush through both stopcocks with
                electrolyte (70$ acetic acid) until the meter reads
                zero or to the left of zero.  (NOTE:   Final level of
                electrolyte should be at black mark on cell.)

           d.   Turn recorder power and chart on and adjust needle on
                recorder to zero with RECORDER ZERO knob on coulometer
                amplifier.  (NOTE:  Needle on recorder is at 0 when
                integrator draws a straight line.)

           e.   Set FUNCTION switch to OPERATE.  Set RANGE OHMS to
                500 or desired setting and vait until cell generates
                as much titrant as it needs to reach null.

           f.   Using a 10 ul syringe, inject a few micrograms of
                standard chloride solution directly into the titration
                cell through the filling funnel.  (NOTE:  Tip of needle
                must be below the level of the electrolyte before
                injection.)

     4.3   Measurement of Area Under Peak

           a.   Each traverse of the integrator pen represents 10 disc
                integrator units (DU).  Find the total number of DU
                under the peak.

           b.   1 DU = rrrrT square inch

                Calculate the area in square inches under the peak.

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66
                                                                                   I
Pesticides in Bottom Sediments,  Microcoulometric Gas Chromatograph


          c.   Calculate the micrograms of chloride represented by                 •
               this area using the following formula:

               UR chloride *  Peak area (in set, in.) x
               ug cnxoriae
          d.   Calculate the recovery of chloride ion in the titration
               cell using the following formula:

               % Recovery        ,   ^g chlofde recovered
               Titration Cell =  (as measured area under p	
                                     ug chloride injected                          •

               If % recovery is below 90%, clean electrodes in NH^OH               •
               and rinse thoroughly with distilled water before                    •
               returning to titration cell.  Check % recovery again
               before injection samples into the gas chromatograph.

     k.k  Measurement of Standard Pesticide Solution                               •

          a.   With RANGE-OHMS at 000 and the vent closed, zero the                •
               recorder.  Increase RANGE-OHMS to 050 or other desired              I
               range-OHMS and wait until cell reaches null.

          b.   With VENT open inject 200 ng of aldrin into the gas                 I
               chromatograph.  Depress MARK button.                                ™

          c.   After one minute, close vent and wait for aldrin peak.              •

          d.   Using the disc integrator, calculate the area under the
               peak.                                                               •

          e.   Calculate the micrograms of aldrin represented by this
               area using the following formula:  (or by calculation
               given under injection of unknown)                                   •
                          , _  Peak Area (in sg. in.) x *4t-2	
               ug  compoun  -  Range OHMS x ($> chlorine in compound)               _


               (Reference  7.6 gives the percentage chloride composition
               of some common chlorinated hydrocarbons.)                           «

          f.   Calculate the recovery of pesticide in the gas chromato-
               graphic system using the following formula:
                                      |ig of pesticide recovered                    I
               nt          j. 4. -, - (as measured by area under peak)                 •
               % recovery total	ug of pesticide injected	L

          g.   Measure peak area using triangulation and the half-width             •
               methods and compare the results with integration.
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                                                                    67


Pesticides in Bottom Sediments, Microcoulometric Gas Chromatograpn



     4.5  Measurement of Unknown


          a.   With RANGE-OHMS at 000 and vent closed, zero the recorder.
               Set RANGE-OHMS to desired ohms and wait until cell reaches
               null.


          b.   With VENT open, inject appropriate amount of unknown.
               Depress MARK button.


          c.   After one minute, close the vent and wait for unknown
               peak(s).


5.   Calculations

               (A unk)           ,
     u£ -ngS'H)(A STP) F. V. x Kr
     kg   (J.1 inj. x g sample


          [>g » Microgram
          ng = Nanogram
          A unk ~ Area Peak of Unknown
          A STD " Area Peak of Standard
          Hi inj. = Microliters extract injected
          g sample = Grams of Sample
          F. V. = Final volume in milliliters

6.   Recovery


     6.1  Weigh 100 g of bottom sediment sample into a biologically
          clean sampling bottle.


     6.2  To 50 ml hexane in a small beaker, add a known amount of
          pesticide standard (in the concentration range of interest).


     6.3  Carefully add the pesticide standard solution (with stirring)
          onto the sample.


     6.4  Allow the solvent to evaporate at room temperature (stir
          periodically; this may take a day or two).


     6.5  When solvent has completely evaporated, transfer sample to
          Soxhlet extractor and carry out procedure for analysis as
          in Section 4.2.


     6.6  Calculate the recovery using the following formula:

          at          - amount of pesticide found by MCGC
          % recovery —   ' * '    '  *       "
          '         J   known amount of pesticide added

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                                                                             I
                                                                    68

Pesticides in Bottom Sediments, Microcoulometric Gas Chromatograph                 •
7.   References

     7.1  Barry, H. C. et. al.,1965.Pesticide Analytical Manual
          (Vol. I revised).  U.S. Department of Health, Education,
          and Welfare, Food and Drug Administration.
                                                                             I
7.2  Breidenbach, A. W. et al., 1966.  The Identification and                 _
     Measurement of Chlorinated Hydrocarbon Pesticides in Surface             •
     Waters.  U.S. Department of Health, Education, and Welfare,              ™
     Public Health Service WP-22 November.

7.3  Hundley, J. C. et. al.,1964.  Pesticide Analytical Manual                I
     (Vol. II).  U. S. Department of Health, Education, and
     Welfare, Food and Drug Administration.                                   •

7.4  Burchfield, H. P. and Johnson, D.E., 1965.  Manual of Methods
     for the Analysis of Pesticide Residues.  Southwest Research
     Institute (Prepared for DHEW, PHS), Vol. I and II.                       •

7-5  Burke, J.  and Giuffrida, A. L., 1964.  Investigation of
     Electron Capture Gas Chromatography for the Analysis of                  •
     Multiple Chlorinated Pesticide Residues in Vegetables.                   |
     J. Assoc. Off. Agr. Chemists 4-7:326.

7.6  Dohrmann Instruments Company, 1966.  Preliminary Operation               •
     Instruction for Dohrmann Microcoulometric Titrating System.              ™
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                                                                    69


                     PESTICIDES IS BOTTOM SEDIMENTS


                        Thin Layer Chromatography



1.   General Discussion


     Thin layer Chromatography (TLC) is one of the most widely used
     analytical and clean up techniques employed in Chromatography.
     Separation of compounds having various polarities, water solubilities
     and having different carbon chain lengths can be separated by TLC.


     1.1  Principle:


          a.   Separation and purification of a chemical mixture is
               effected in thin layer Chromatography by forced move-
               ment of a spot of material with the aid of a suitable
               migrating solvent through a thin layer of sorbent
               material which has been applied to an appropriate
               support.  The mechanism of separation is one or more
               of the following:  adsorption, partitioning or reversed
               phase partitioning.


          b.   Adsorption is useful for the separation of lipophilic
               organic compounds of low or medium polarity.  The
               adsorption material used is silica gel or aluminum oxide.


          c.   Partitioning is useful for the separation of water soluble,
               inorganic compounds or quite polar organic materials.
               (Paper was formerly used, but now, cellulose crystalline
               powdsror ground fibers is being used; although unactivated
               silica gel, carrying an absorbed phase of water can be
               used.)


          d.   Reversed phase partition TLC is good for resolving closely
               related lipophilic materials like an homologous series
               differing only in carbon chain length.


2.   Apparatus


     2.1  Plates, 200 mm x 200 mm (8" x 8") glass.


     2.2  Chamber, glass developing, with lid, 8%" x V x 8^-".


     2.3  Spreader, variable thickness.


     2.4  Spotting template.


     2.5  Chromatography sprayer.

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                                                                    70             I


Pesticides in Bottom Sediments, Thin Layer Chromatography                          •


     2.6  Desiccator to accommodate 200-mm plates.                                 I

     2.7  UV light box.

     2.8  Micropipets, lul-5nl- lOu.1, 100|al.                                        |

     2.9  Eye droppers.                                                            _

     2.10 Spatulas.                                                                •

     2.11 Graduated centrifuge test tubes, capacity 15 ml.                         •

     2.12 Glass wool, pre-extracted with chloroform.

     2.13 Centrifuge.                                                              I

     2.Ik Cylinder, 100 ml.

     2.15 Drying rack to hold 8" x 8" glass plates.                                •

     2.16 Volumetric flask, 10 ml.

3.   Reagents

     3.1  Pesticide standards.

     3.2  Khodamine B base, spirit soluble 0.10 rag/ml in ethanol.

     3.3  Aluminum Oxide G.                                                        •

     3.4  All solvents are redistilled before use.                                 •

     3.5  Hexane: ethyl ether; 4:10 mixture.

     3.6  Carbon Tetrachloride.                                                    •

     3.7  Hexane.

k.   Procedure                                                                     •

     4.1  Preparation of Plates                                                    •

          a.   Weigh a 40 gram aliquot of Al20o in a 500 ml centrifuge
               bottle and add 80 ml 0.03 W HWO^.  Shake well and centri-
               fuge at 1200 rpm for 1-2 mTnutes.                                   •
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                                                                   71

Pesticides in Bottom Sediments,  Thin Layer Chromatography
          b.   Decant the supernatant liquid into  a 100 ml  graduated
               cylinder,  and record the volume  (35-^0 ml  should be
               recovered).


          c.   Add 80 ml  distilled 1^0,  break up cake on  the bottom
               of the bottle with a glass  rod,  shake well and  centri-
               fuge as before,  and decant  and record the  volume of
               the supernatant  liquid recovered (70-80 ml).


          d.   Repeat the washing step with  two additional  80  ml
               portions of HgO.   Centrifuge  and decant as before.


          e.   Weigh the  A^O,  and H/jO which has been retained
               (approximately 100 g).


          f.   Add 10 ml  1.0$ AgN03 and enough  1^0 to make  the total
               weight 120 g,  and shake well,  place in applicator and
               prepare plates.

          g.   Let plates dry on mounting  board for approximately
               15 minutes.

          h.   Place in a drying rack in a vertical position at 100°C
               for 30 minutes.

          i.   Remove the plates and store in a desiccator.


          Spotting Plates


          a.   Position plate on Kimwipe or  clean  surface and  scrape
               1/V area  on two  sides of plate.


          b.   Insert (5  (il)  capillary spotting  tube in small  beaker
               of hexane  and touch to folded Kimwipe until  all hexane
               has been removed  from capillary.  Repeat this five times.

          c.    Place template with graduated edge  over the  aluminum
               oxide AgKOo impregnated plate about 1" from bottom.

          d.    Mark this  position with pencil on both sides of plate.


          e.    Rinse capillary twice with  standard pesticide solution
               removing contents each time on kimwipe.

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                                                                    72


Pesticides in Bottom Sediments, Thin Layer Chromatography                           I


          f.   At graduation 1 of template, touch capillary filled
               with standard once lightly on plate removing approxi-
               mately 1/4 of contents of capillary (plate position #l).

          g.   At position #2, spot two times putting 1/2 the contents              I
               of the capillary on the plate.  Permit each spot to dry
               before respotting.
     4.4  Identification and Estimation Using Rhodamine B
                                                                                    I
     h.   At position #3,  spot the contents of the capillary (5i-il)
          standard by lightly touching the capillary to the plate,
          removing about 1/4 of the contents with each spotting.
          Permit the spots to dry before respotting.

     i.   At positions #4 - #6 spot sample labelled "unknown,"
          repeat spotting procedure as described in items f - h.              •
          Repeat above procedure for other pesticide solutions and            •
          standards.

4.3  Spot Development                                                         |

     a.   Place plate, coated surface facing solvent saturated paper,         _
          in developing tank containing 1 cm depth of carbon tetra-           •
          chloride with marked spotting line down; develop to the
          10 cm line.

     b.   Lift off top of jar, mark solvent front on side of plate            •
          and remove plate, lifting plate with finger tips of both
          hands.  Dry in air for 10 minutes.                                  •

     c.   Place in an ultraviolet light box and expose for 15 minutes.

     d.   Measure the distance of centers of the spots from starting          •
          line and compute the R-p values.  Record data on data sheet.         ™
                                                                              I
     a.   Repeat spotting procedure described above employing aluminum
          oxide plain plates.  Spot the standards and bottom sediments        •
          extract sample and develop in solvent tank.                         I

     b.   Place plate in hood and spray with Rhodamine B 0.1 g/1 in
          ethanol.  The spray should be a fine mist giving an even,           •
          smooth lifirht r>ink color to surface of ulate.                        B
                smooth  light  pink color  to  surface  of plate.
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                                                                    73

Pesticides in Bottom Sediments, Thin Layer Chromatography



          c.   Place plate in dark and observe spots with UV light.


          d.   Measure distance (cm) of spotting line to solvent front.
               Measure distance of spots from starting line.

5.   Calculations


     5.1  Calculate Rj, of spots which appear in samples.

               R    Distance of spot from starting line	
                f * Distance of solvent front from starting line


     5.2  Identify unknowns by comparing R,, value with standards.


     5-3  Estimation of concentration in unknown spots may be done
          by comparing with standard pesticide spots.


6.   Preparation of Thin Layer Spots for Gas Chromatography


     6.1  Marking off Spot Area on Plate


          a.    Place template below lower spot line.  Mark surface with
               spatula across plate.


          b.    Move template above these spots and similarly mark  surface.


          c.    Move template to a line above the pesticide spots and mark
               surface.


          d.    Select the lowest concentration of pesticide spots  showing
               best color development,  such as,  1.25, 2.50, 5-0 or 10 |al
               spots.


          e.    Turn template perpendicular to selected spot;  mark  surface
               across plate.
•                     f.   Move template to other side of spot and mark surface.

                  6.2  Removal of Spot from Plate
          a.    Insert a tuft of glass wool and position at the tapered
               end of medicine dropper.


          b.    Mark the medicine dropper to identify the pesticide spot
               collected.

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                                                                    7-             I


Pesticides in Bottom Sediments, Thin Layer Chromatography                          •
               Connect vacuum tubing over tapered end of medicine dropper.
     Administration.
I
          d.   Collect all of adsorbent surface enclosed between boundaries
               of marked off area by sucking the adsorbent and pesticide           •
               by vacuum and depositing in glass wool of medicine dropper.         I

     6.3  Elution of Pesticide

          a.   Mark a 10 ml volumetric flask according to the pesticides           •
               studied.

          b.   Place paper clip over tapered end of medicina dropper               |j
               approximately 2/3 distance from large end.

          c.   Insert tappered end of dropper into volumetric flask                •
               supported by paper clip.

          d.   Elute pesticide with 5 nil of k:i hexane-ether mixture.              I

          e.   Fill to mark with hexane.

          f.   The elute is now ready for further identification by                |
               gas chromatography.

7.   Reference                                                                     •

     Barry, H. C., et. al. 1965 Pesticide Analytical Manual (Revised Vol.1).
     U.S. Department of Health, Education, and Welfare, Food and Drug              •
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                                                                    75


                   PESTICIDES IK BOTTOM SEDIMENTS


                              Infrared



1.   General Discussion

     This method is suitable for mg amounts; but with beam condenser
     or internal reflectance accessories, ng quantities may be analyzed.


2.   Apparatus


     2.1  Twelve ton press with a KBr pellet maker.


     2.2  Agate mortar and pestle - approximately 4-0-75 mm OD.


     2.3  I. R. spectrophotometer with beam condenser, internal
          reflectance, and scale expansion.


3.   Reagents


     Potassium bromide, infrared quality.


4.   Procedure


     4.1  Grind 50-100 mg of KBr (dried overnight at 150°C) and mix
          50 ng of sample or standard.  (This may be done under IR lamp.)


     4.2  Press into KBr-disc.  (Disc should be clear.)


     4.3  Dry 2 hours in a vacuum oven at 50 °C and 1 mm/Hg.


     4.4  Place disc in spectrophotometer and record IR spectrum.


     4.5  Run standards in the same manner.

5.   Calculations


     Compare IR fingerprints with standards for positive identification.

6.   References
|               6.1   Breidenbach, A.W., et.al. 1966.  The Identification and Measurement
                      of  Chlorinated Hydrocarbon Pesticides in Surface Waters.  U.S.Dept.
I                      of  Health,  Education, and Welfare, Public Health Service WP-22
                      November.
     6.2  Burchfield,  H.P.  and Johnson,  D.E.,  1965.   Manual of Methods for
          the Analysis of Pesticide Residues.   Southwest Research Institute
          (Prepared for DHEW,  PHS),  Vol. I and II.

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                                                                                     I
Pesticides in Bottom Sediments, Infrared
I
     6-3  Parkin-Elmer Corp., 1967.  How to Collect Fractions from the               —
          Model 800 Series Gas Chromatograph.  GC Newsletter, Vol.. 3,                I
          No. 1, Norwalk, Connecticut.

     6.if-  Sadtler, T. and P. Sadtler, 1965.  Improved KBr Techniques                 •
          presented:  Pittsburg Conference on Analytical Chemistry and               •
          Applied Spectroscopy, Sadtler Research Labs., Inc., 331^ Spring
          Garden Street, Philadelphia, Pa.                                           •




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                                                                 77
                     PHENOL IN BOTTOM SEDIMENTS


1.  General Discussion


    1.1  Principle:  Phenolic types of compounds but not paracresol
         and similar parasubstituted phenol's react with amino anti-
         pyrine at a pH 10.0 ± 0.2 in the presence of ferricyanide
         to form an antipyrine dye.  The dye is extracted with
         chloroform from the aqueous solution and the absorbance
         read at h60 m^i •


    1.2  Interference:  Most interferences are eliminated or reduced
         to a minimum by using the distillate of the original sample.


    1.3  Minimum Detectable Concentration:  The minimum detectable
         quantity is 0.5 microgram with a 25 ml extraction and when a
         5 cm cell is used in the  photometric measurement.


2.  Apparatus


    2.1  Distillation apparatus, all glass.  A suitable assembly
         consists of a 1 liter pyrex distilling apparatus with Grahm
         condenser.


    2.2  Separatory funnels,  Squibb form with teflon stoppers 1,000 ml
         capacity.


    2.3  Spectrophotometer.


3«  Reagents


    3.1  Phenol Stock Solution:  Dissolve 1.0000 g of phenol in
         distilled water and dilute to 1,000 ml with distilled water.
         1 ml s 1 mg phenol

    3.2  Phenol Working Solution:  (Must be prepared daily)  Pipette
         20.0 ml of stock solution into a 1 liter volumetric flask
         and dilute to volume with distilled water.   1 ml =  0.020 mg


    3.3  Phenol Standard Solution:  Pipette 25.0 ml  of  working
         solution into a 500 volumetric flask and dilute to volume
         with distilled water.  1 ml =  0.001 mg


    3.^  Ammonium Chloride Solution:  Dissolve 67.5  g of NH^Cl in
         570 ml of  cone.  NH^OH and dilute to one liter  with distilled
         water.


    3»5  Aminoantipyrine Solution:  Dissolve 2.0 g of k- - amino-
         antipyrine in distilled water and dilute to 100 ml volume.


    3.6  Potassium  Ferricyanide  Solution:  Dissolve  8.0 g KoFe(CN)6
         in distilled water and  dilute to 100 ml.

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                                                                      73
Phenol in Bottom Sediments

       3.7  Chloroform:  Reagent grade of chloroform (redistilled).                    •

       3.8  Phosphoric Acid Solution:  Mix 10 ml of phosphoric acid
            with distilled water and dilute to 100 ml.                                 •

       3.9  Copper Sulfate Solution:  Dissolve 10 g of CuSO^.^O in                   •
            distilled water and dilute to 100 ml.

  h.   Procedure                                                                       I

       U.I  An aliquot of sample containing not more than £0 * ng of                     —
            phenol is placed in the distilling flask.  (20 g)                           g

       lj.2  Add 550 ml of distilled water.

       h.3  If the sample has not been preserved, add 5 ml of 10£ CuSO^                |
            and phosphoric acid and a few drops of methyl orange indicator.

       luU  Add a few boiling chips and distill over 500 ml of distillate.             •


            k.ka If oil is present in distillate, filter through two thick-            •
                 nesses of dry No. 12 Whatman filter paper into separatory              •
                 funnel to remove oil.

        ii.5  Transfer the distillate to a one liter separatory funnel,                 I
             add 3 ml of  NHj^Cl and nix, 3 ml of aminoantipyrine and mix,               ™
             and 3 ml of  potassium ferricyanide and mixe   Let  stand at
             least 3  minutes o                                                           •

        U.6  Add 25 ml of CHClo and  shake vigorously for 30 seconds:
             after the layers  have separated,  shake once more  for 30 seconds.           _

        ll.7  When the layers have  separated,  draw the CHC1., layer off                  ™
             through  a cotton  pledget placed  in the stem of the separatory
             funnel into  a 25  ml graduate  cylinder.  (Do not make to  volume.)           •

       U.8   Read the optical  density at  a wave length of Ij60  n^i.

       ii,9   Standards and blanks  are treated exactly as the sample.                    I

  5»   Calculations

       5.1  Wet basis

                      e i" std     ..                                                  _
                                 grams Sample                                          •
    mg/kg =  -n-e i" std. x ().D. Sample x 1000
             00Do Stdo
5.2  Dry basis

          B mg Ag wet basis _                                       I
              % Solids (decimal fractio^J

                                                                                I
       Reference

       Standard Methods for the Examination of Water and Wastewater, 12th
       Ed., APHA, Inc., N.Y., 1965, 51^-520.
                                                                                 •

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                                                                 79


           TOTAL SOLUBLE PHOSPHORUS Df BOTTOM SEDIMENTS



1.  General Discussion


    1.1  Principle:  Ammonium molybdate and potassium antimonyl tartrate
         react in an acid medium with dilute solutions of ortho-
         phosphate to form a heteropoly acid (phosphomolybdic acid)
         which is reduced to the intensely colored molybdenum "blue by
         ascorbic acid.


    1.2  Interferences:  Arsenates react with the molybdate reagent to
         produce a blue color similar to that formed with phosphate.
         Concentrations as low as 0.10 mg/1 of arsenic interfere with
         the phosphate determination.  Hexavalent chromium and nitrite
         interfere to give results about 3$ low at concentrations of
         1.0 mg/1 and 10-15$ low at concentrations of 10 mg/1
         chromium and nitrite.  Sulfide (Na2S) and silicate do not
         interfere in concentrations of 1.0 and 10 mg/1.


    1.3  Precautions:  All glassware should be washed with hot 1:1
         hydrochloric acid and then rinsed thoroughly with distilled
         water.  The glassware is then filled with distilled water
         and all reagents added and allowed to stand 15 or 20 minutes,
         followed by a distilled water rinse.  Commercial detergents,
         should not be used.


2.  Apparatus


    Colorimetric equipment:  A spectrophotometer with an infrared
    phototube for use at 880 m^t, providing a light path of 1" or longer,
    should be used.


3.  Reagents


    3-1  1.0 N H2SO^.

    3-2  10 N NaOH.



    3-3  1.0 N NaOH.


    3A  Sulfuric  Acid,  8N:   Dilute 112 ml of cone.  H2SO,  with
         distilled water to 500 ml.

    3-5  Potassium Antimonyl Tartrate:   Dissolve 4.3888 g K(SbO)C. H, 0^«
         1/2 HJD in 200 ml  of distilled water.   Store in dark bottle
         at 4°&.

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                                                                    80                |

   Total Soluble Phosphorus In Bottom Sediments


                                                                       i .- _           Hi
       3.6  Ammonium Molybdate Solution:  Dissolve 20 g.
            in 500 ml of distilled water.  Store in plastic bottle'al
            4°c.                                                                      I

       3.7  Ascorbic Acid,  0.1M:  Dissolve 1.76 g:   of ascorbic acid in
            100 ml of distilled water.  The solution is stable for about              •
            a week if stored at 4°C.                                                  I

       3.8  Combined Reagent:  Mix the above reagents in the following                •
            order and proportions for 100 ml of the combined reagent:                 •
            to 50 nil of 8N I^SOj^  add 5 ml of potassium antimonyl tartrate
            solution and mix, then add 15 ml of ammonium molybdate solution
            and mix,  and finally, 30 ml of ascorbic acid solution and mix.            •
            All reagents must be  at room temperature before mixing.   The              •
            combined reagent should be prepared fresh each day just before
            use.                                                                      •

       3.9  Alcohol,  ethyl (95 per cent) or isopropyl.

       3.10 Stock Phosphorus Solution:  Dissolve in distilled water 2.1968 g:          I
            of potassium dihydrogen phosphate, K^FOjp which has been dried
            in an oven at 105*C.   Dilute this solution to 1,000 ml.  One ml
            equals 0.50 mg P.                                                         I

       3.11 Working Phosphorus Standard #1:  Dilute 20 ml of stock phosphorus
            solution to 1,000 ml  with distilled water, 1 ml equals 0.01 mg P.         •

       3.12 Dilute 100 ml of working standard #1 to 1,000 ml with distilled
            water, 1 ml equals 0.001 mg P.                                            »

       3.13 Strong Acid Solution:  Slowly add 3OO ml cone. I^SO^ to 600 ml            ™
            distilled water.  When cool, add 4.0 ml cone. HNO^ and dilute
            to 1 liter.                                                               •

       3.l4 Potassium or ammonium Persulfate, analytical grade.

       3.15 Concentrated HNOo.                                                        J

4.     Procedure

       4.1  Place 5 g of well blended sample into a 100 ml beaker.  Add               ™
            50 ml of distilled water and mix.  Let this solution set
            overnight at room temperature.                                            •

       4.2  Filter the above mixture through a membrane filter
            0.45n porosity and place entire filtrate in a 250 ml beaker.              M
            Do no wash residue on filter.                                             I

       4.3  Add 1 ml of strong acid and 0.4 g:   of potassium persulfate
            (K0S0Oft).  Boil gently for 90 minutes, adding distilled water             •
            if necessary to keep the volume between 25 and 50 ml.
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                                                                    81
Total Soluble Phosphorus in Bottom Sediments
                  k.k  Cool, neutralize to a faint pink color with IN NaOH and
mm                     transfer to 100 ml nessler tubes.  Dilute to volume.
                  4.5  Add 5 ral of ethyl or ispropyl alcohol and mix thoroughly.
     4.6  Add 5 nil °f combined reagent.  Mix thoroughly and allow to
          stand 10 minutes for color development.  Read at 880 v\i.
     4.7  A standard and blank should also be run with each analysis.
5.   Calculations
     5.1  Wet Basis
                  mg in Std.   P.P. Sample     ioon
                r O.D. Std.  x (grams sample x
                                in aliquot)
     5.2  Dry Basis
                  mg/kg vet basis	
                       mg/kg
                  % solids (decimal fraction)
     References
     6.1  "Methods for the Collection and Analysis of Water Samples,"
          Geological Survey Water Supply Paper 1454, I960, pp 245-250.
I
                  6.2  Official Methods of Analysis, A.O.A.C., 7th Ed., 1950, p 370.
I                6.3  Journal AWWA, July 1965, PP 917-925.
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                                                                  82

                    TOTAL PHOSPHORUS IN BOTTOM SEDIMENTS
3.5  10$ Na2S03:   Dissolve 100 g Wa2SOo (phosphate free)  in 1,000 ml
     of distilled water.  Prepare fresn daily.
                                                                                   I
1.  General Discussion

    1.1  Principle:   Ammonium molybdate and hydrazine sulfate react                 I
         in an acid medium with dilute solutions of orthophosphate to
         form a heteropoly acid (phosphomolybdic acid) which is reduced             •
         to the intensely colored molybdenum blue by sodium sulfite.                 •

    1.2  Interferences:   Arsenic,  iron,  tungsten, silica,  titanium,
         zirconium and vanadium do not interfere.                                   •

    1.3  Precautions:  All glassware should be washed with hot 1:1
         hydrochloric acid and then rinsed thoroughly with distilled                •
         water.  The glassware is then filled with distilled water                  |
         and all reagents added and allowed to stand 15 to 20 minutes,
         followed by a distilled water rinse.  Commercial detergents                 .
         should not be used.                                                        •

2.  Apparatus

    Colorimetric equipment:  A spectrophotometer with an infrared                   I
    phototube for use at 810 mp., providing a light path of 1" or longer
    should be used.                                                                  •

3-  Reagents

    3.1  3$H202.                                                                   I

    3.2  Perchloric acid •

    3.3  50$ Mg (NO )2 • 61^0:  Dissolve 500 g Mg (N03)2 . 6H20 in 1,000 ml         |
         of distilled water.

    3-^-  Concentrated hydrochloric acid.                                            •


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    3.6  0.15$ Hydrazine Sulfate:   Dissolve 1.5 g of the salt in 1,000 ml
         of distilled water.  Prepare fresh daily.                                  •

    3-7  Ammonium Molybdate:  Add 300 ml of cone. ^SOlj. to 500 ml of
         distilled water and cool to room temperature.  Dissolve 20g of             _
         ammonium molybdate in the ^SO^ solution and dilute to 1,000 ml            •
         with distilled water.  (Do not use the regular 1:1 - H2SO^ for             ™
         this - mix as directed.)
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                                                                  83

Total Phosphorus in Bottom Sediments

    3.8  Mixed Reagent:  Dilute 25 ml of ammonium molybdate solution
         with 45 ml of distilled water.  Add 10 ml of the hydrazine
         sulfate solution and 20 ml of the sodium sulfite solution,  and
         dilute to 100 ml with distilled water.  The reagent is stable
         for about one-half hour.

4.  Procedure

    4.1  Weigh 1.00 g of well-blended sample into a 250 ml Vycor dish.

    4.2  Add 10 ml of 3% H202 and heat on hot plate for 10 or 15 minutes
         with stirring to prevent overflowing from frothing.

    4.3  Add 10-15 ml of distilled water and continue to boil until
         frothing has ceased and almost all of the water has evaporated.

    4.4  Add 10 ml of 50$ Mg (NO-^^I^O and place in a cold muffle
         furnace.  (Keep stirring rod in Vycor dishes and make sure
         dishes are permanently marked.)

    4.5  Start at 100°C and increase the heat gradually until the
         samples are dry (about 350°C).  Then increase heat to
         500-550°C and ash for one hour.

    4.6  Remove samples from furnace and let cool on asbestos pads.

    4.7  When cool,  add 20 ml of cone. HC1,  place on hot plate and
         heat with stirring until effervescence has stopped.

    4.8  Add approximately 50 ml of distilled water and clean the
         sides of the dish with a rubber policeman.

    4.9  Filter through membrane filter 0.45n into a 250 ml
         volumetric  flask,  wash dish and filter paper with distilled
         water and make up to volume.

   4.10  Take a 25 ml aliquot, add 4 ml perchloric acid and fume until
         fumes reach the mouth of the flask.

   4.11  Cool, then add 10 ml of distilled water, 15 ml of 10$ Na2SOo
         and boil for at least one minute, but not longer than two
         minutes.

   4.12  Cool, then add 20 ml of the mixed reagent and heat in a boiling
         water bath for 15 minutes and cool rapidly to 20°C.

   4.13  Transfer to a 100 ml nessler tube and make up to volume with
         distilled water and read at 810 up against distilled water.
         The usual blank for reagents is 0.015.

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 Total  Phosphorus  in Bottom Sediments                                               I


5-  Calculations

    5-1   Wet Basis                                                                 •
                               X V>L>> samvi.e   „ . oon                              •
                  O.D. Std.      (grams sample A ^U                                I
                                  in aliquot)

    5.2   Dry Basis                                                                 •

             /   _ mg/kg wet basis	
          "•S/KS - 56 solids (decimal fraction)

6.  References                                                                      •

    6.1  "Methods for the Collection and Analysis of Water Samples,"                •
         Geological Survey Water Supply Paper 1^54, I960, pp 2^5-250.               |

    6.2  Official Methods of Analysis, A.O.A.C., 7th Ed., 1950, p  370.              _

    6.3  Journal AWWA, July 1965, pp 917-925.                                       "

    6.k  Chemical Analysis of Iron and Steel, Lindell, Hoffman and Bright,          I
    6.5  U.S. Bureau of Mines, Minneapolis, Minnesota,
         Personal Communication.
         1931, PP 209-231.

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85
SOLIDS - TOTAL AND VOLATILE IN BOTTOM SEDIMENTS

1. General Discussion

1.1


1.2



1.3


Principle: The sample is dried in a weighed dish in an oven
at 103-105>°C to constant weight. The increase in weight over
that of the empty dish represents the total solids.
The volatile solids are determined by placing the oven dried
sample from 1.1 in the muffle furnace for one hour at 600°C.
The decrease in weight of residue after ashing represents the
volatile solids.
Minimum detectable concentration: Dependent on the sensitivity
of the analytical balance used for weighing.
2 . Apparatus
2.1
2.2
2.3

2.L
2.5
Drying Oven.
Muffle Furnace.
Porcelain Crucibles .

Porcelain Evaporating Dishes .
Analytical Balance.
3. Procedure

3.1
3.2

3.3


3.1
3.5

3.6
3.7


3.8
3.9



Wash dishes and number with heat-resistant marking pencil.
Place evaporating dishes in a muffle furnace at a temperature
of 600-6^ 0°C for one hour.
Remove evaporating dishes from the furnace and allow them to
cool for 1-2 minutes in air, but not more than 3-U minutes.
Then place them in a dessicator for 1 houra
Weigh and record this weight as the tare weight.
Weigh 10 grams of bottom sediment to the nearest 0.01
grams in the tared evaporating dishes .
Place the sample in the oven at 103-10£°C overnight.
Remove samples from the oven and place them in a dessicator
for 1 hour.

Weigh and record this weight as the oven-dry weight.
Place the oven dried sample in the muffle furnace at 600°C
for 1 hour0


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                                                                  86

Solids - Total and Volatile in Bottom Sediments
     3.10 Remove samples from the furance and allow them to cool
          for 1-2 minutes in air, but not more than 3-U minutes.
          Afterwards, place then in a dessicator for 1 hour.
     Standard Methods for the Examination of Water and Wastewater,
     12th ed., APHA, Inc., N.Y., 1Q65, 53^-535-
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     3.11 Weigh and record this weight as the furnace weight.                     _

U.   Calculation

     h.l  Oven dry weight x 100 = % total solids                                  •
          Initial wt. of. sample                                                   I
     U.2  Oven dry weight - furnace weight x 100 = % volatile solids
               Oven dry weight                                                    •


£•   Reference
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                                                                87
            TOTAL SULFIDE IN BOTTOM SEDIMENTS
1.  General Discussion

    Total sulfide includes dissolved hydrogen sulfide and its ionization
    products as well as acid-soluble metallic sulfides present in sus-
    pended matter.  Sulfide ion is also determined by the following
    method, but it is not present in significant concentrations below
    a pH of 13.

    1.1  Principle:  Sulfides are removed from suspended matter and
         other interfering substances in solution by distillation
         under acidic conditions.  Air oxidation of the evolved
         hydrogen sulfide is minimized by passing nitrogen gas
         through the system which displaces the oxygen prior to
         acid addition.  The distilled hydrogen sulfide is collected
         in zinc acetate which reacts to form insoluble zinc sulfide.

         The zinc sulfide is reacted with N,N-Dimethyl-p-phenylene-
         diamine in sulfuric acid to produce an intermediate that is
         oxidized by ferric chloride to an intensely blue colored dye
         called methylene blue.  The intensity of the blue color is
         proportional to the original concentration of sulfide in the
         distillate and is measured at a wave length of 650 rn^i.

    1.2  Sampling and storage:  Samples must be taken with a minimum
         of aeration, for not only are sulfides volatilized by aeration
         but also any oxygen that is taken up will readily oxidize the
         sulfides to free sulfur.

         The sample is preserved by pipetting 2 ml of 2N zinc acetate
         solution into the bottle, replacing the stopper, and shaking
         well.  This precipitates the sulfides as inert zinc sulfide,
         which not only prevents oxidation of sulfides but also
         prevents further sulfide generation.

2.  Apparatus

    2.1  Distillation apparatus, all glass.  For large samples, a
         suitable assembly consists of a 1-liter pyrex distilling flask
         with Graham condenser as used for the analysis of phenols.

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                                                                              I
Total Sulfide ±n Bottom Sediments                                    A«
                                                                    oo

          A section of glass tubing should be connected to  the tip  of  the          |
          condenser so that it reaches the bottom of the collection tube.

     2.2  Distillate collection tubes.  Short form Nessler  tubes.                  I
          graduated at 50 and 100 ml.                                             ™

     2.3  Spectrophotometer, for use at 650 mn and providing a light              I
          path of 1 Inch or greater.                                               •

3«   Reagents                                                                     •

     3»1  Nitrogen, water-pumped.
 3.2  Zinc acetate,  2N.   Dissolve 220 g of Zn ^H^^.SHgO in               g
      distilled water and dilute to 1 liter.

 3.3  Zinc acetate,  0.2M.  Add several drops  of  acetic  acid  to 100 ml         I
      of 2N zinc acetate solution and dilute  to  1 liter.                      •

 3.U  Sulfuric acid  solution,  1:1.  Add,  cautiously, 500 ml  of cone.          •
                                                                             I
                to 500 ml of distilled water in a 1-liter flask.  Mix
          continuously and cool under running water while combining
          reagents.  Cool solution before using.                                 •

     3.5  Dilute sulfuric acid solution,  approx. 0.1N:   Dilute  5  ml  of
          1:1 H2SOk to 1 liter with distilled water.                              _

     3.6  Stock amine solution:  Dissolve 2.7 g of N,N-Dimethyl-p-
          phenylenediamine sulfate and dilute to 100  ml with 1:1  H^SO.
          solution.  This solution is stable for approximately  one                •
          week.                                                                  •

     3.7  Working amine solution.  Dilute 2 ml of stock amine solution            •
          to 100 ml with 1:1 H2SO^ solution.  Prepare fresh  daily.                |

     3.8  Ferric chloride solution:  Dissolve 100 g of  FeCl.,.6H20 in
          hot distilled water and dilute  to 100 ml.  Cool before  use.
                                                                             I
 3.9  Standard potassium biniodate solution,  0.025N.  Accurately
      weigh out 0.812U g KH(IOo),., and dissolve in  distilled water.            I
      Dilute to 1-liter.                                                     •

3.10  Standard sodium thiosulfate titrant,  0.025N. Dissolve  6.205  g          I
      N2S2°3^H2° ^ distilled water and dilute to 1  liter.                   •
      Preserve with 5 nil chloroform.  Standardize  against  standard
      potassium biniodate using starch as an  indicator.                       •
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Total Sulf lie in Bottom Sediments                                    89


     3.11 Potassium iodide solution.  Dissolve 5 g of KI in distilled
          water and dilute to ICO ml.

     3.12 Treated hydrochloric acid.  Place one or two strips  of  aluminum
          in a small beaker of concentrated HC1.  After violent reaction,
          the acid is poured off and is ready to use.

     3.13 Oxygen-free dilution water.  Pass nitrogen gas through  a suf-
          ficient quantity of distilled water for dilution requirements.
          A minimum of 10 minutes is required to displace oxygen  in
          the water.

     3.ill Sodium sulfide, Reagent, crystal.

It.   Procedure

     lul  Standardization:  Sulfide solutions are extremely unstable
          and must be prepared fresh and used immediately.   Stability
          is increased by using nitrogen-saturated water for dilution,
          etc.

          Prepare 0.01N sulfide solution as follows:   Weight out  ap-
          proximately 1.2 g of large crystal Na2S.9H20.   Wash  the
          crystals several times with distilled water.  Discard the
          washings and add the washed crystals to 97$ ml of nitrogen-
          saturated distilled water.  Dilute to 1 liter. The  exact
          concentration of this stock solution is determined by
          reacting the sulfide with an excess of iodine  to  give free
          sulfur and titrating the unreacted iodine with sodium thio-
          sulfate.

          Pipet 20.00 ml of stock sulfide solution into  ICO ml of
          oxygen-free water.  Add 5 ml of KI solution, 20.00 ml of
          0.025N KH (103)9 solution, and 10 ml of O.BT HgSO^.  Titrate
          with 0.025N Ife^S^O^ solution using starch as an end point
          indicator.  Carry a blank through the procedure and calculate
          the amount of reacted iodine from the difference between the
          blank and standard titrations.  Since 1 ml  of  0.025N KH(lOo)2
          is equivalent to O.^OO mg of sulfide ion, calculate the
          sulfide concentration in the stock solution.   Calculate the
          ml of stock solution equivalent to 2 mg sulfide and add this
          amount to 9CO ml of oxygen-free water and dilute to 1 liter.
          This is the working standard containing 2 ug sulfide per ml
          of solution.

          Prepare a standard curve using the working sulfide solution
          in the range from 2 to 50 tig sulfide,  as follows:  Pipet
          2O ml of Oe2N Zn (C2H-,02)2 into a series  of 50 ml nessler
          tubes.  Add the required amounts  of sulfide solution to each

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                                                                    90
Total Sulf ide in Bottom Sediments
     tube and dilute to 5>0 ml with oxygen-free waber.   Develop
     the color as outlined In section Iu3 and read the optical
     density at
     50 mis,   Return the transfer tube to the solution and proceed
     with  color development,
                                                                               I
     Prepare a standard curve for distilled samples  by running
     consecutive amounts of the working standard through the  distil-            I
     lation procedure in section U.2.  Determine the sulfide  concen-            •
     tration of each standard run through this procedure by adding
     an identical amount of working standard to 20 ml of zinc                  •
     acetate, diluting to 5>0 ml, developing the color and comparing             |
     the optical density with the standard curve. This step is
     necessary since the sulfide concentration in the working                  •
     standard decreases with time.                                             J

U.2  Distillation Procedure.  Set up the distillation apparatus.                _
     The transfer tubs frcm the condenser should reach to the                  •
     bottom of the distillate collection tube.  The  condenser                  •
     should be attached in such a manner that it can be easily
     moved up or do'.m when diluting the distillate or adding                    I
     reagents .                                                                 •

     Pipet 20 ml of 0.2N ZnCCgH^O^ into a 100 ml nessler tube                •
     and lower the condenser so that the transfer tubing reaches                g
     below the level of the liquid.  Attach a distilling flask
     and pass nitrogen gas through the system for at least 10                  •
     minutes.  Add an aliquot of the bottom sediment  sample contain-             •
     ing not more tfcaa 50 ng of  sulfide.  Bubble nitrogen gas  through
     the sample to remove  any oxygen  dissolved in the sample.   A small
     amount of sulfide may be driven  over by the gas, so be sure that  the       I
     only  exit is  through  the zinc acetate solution in the collecting          •
     tube.   Discontinue nitrogen  evolution and add rapidly several
     boiling stones j  2 drops of methyl orange indicator, and enough            •
     treated HC1 to  change the  color from orange to  red.  Stopper              |
     as quickly as possible and heat slowly   The slower the heating
     rate  the  greater the  contact time between the evolved I^S and             •
     Zn (62^02)2 and the less chance of sulfide loss.  Distill the             I
     solution  until  approximately 20 ml of distillate have been col-
     lected (roughly 5-8 minutes   after the solution commences to
     boil).  Turn  off heat and  remove the stopper in the distil-               I
     lation  flask  to keep  the distillate from being sucked back                •
     up the  condenser.  Raide the transfer tube above the 50 ml
     mark  on the collection container and dilute the solution to               •
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                                                                 91
Total Sulfide in Bottom Sediments
    4.3 Color Development.  Since some zinc sulfide adheres to the
        transfer tube, it should be in the solution while the color
        reaction is taking place.  Furthermore, since this reaction
        is sensitive to temperature, the solution should be immersed
        in a vater bath at 23-25°C while reagents are being added.
        Add 2 ml of dilute amine solution to the 50 ml of distillate,
        mix and add 5 drops of FeClo solution.  Mix again and set
        aside for 10 minutes while the color develops.  Measure the
        absorbance at 650 mjj..


5-  Calculation


    5.1 Wet basis:


        mg/kg e mg in Std» « P.P. Sample  x 1000
                O.D. Std.   (grams Sample
    _ . _                    in aliquot)
    5.2 Dry basis:

              mg /kg wet basis	
               % Solids (decimal fraction)


6.  References


    6.1 Hiotometric Determination of Sulfide and Reducable Sulfur
        in Alkalies, M. Budd, H. Bewick, Anal. Chem. 2^, 1536 (1952).


    6.2 Standard Methods for the Examination of Water and Wastewater,
        12th ed., APHA, Inc., N.Y., 1965,
    6-3 A Revised Method for the Determination of Total Sulfide in
        Water and Bottom Sediments. T. 0. Meiggs, Lake Michigan
        Basin Office,  Chicago,  111.  (1966).

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                                                                    92


                          SILICA IN BOTTOM SEDIMENTS
1.  General Discussion
    1.   Sodium carbonate fusion to decompose refractory silica
         and other such materials.
    5-   Calculation of the amount of silica from the weight lost
         or volatilization.
2.  Apparatus

    2.1  Porcelain evaporating dishes.

    2.2  Porcelain crucibles and covers.
         Pestle, large.
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Although silicon constitutes approximately 26% of the lithosphere,
it is never found in the native state.  As the silicate, it occurs
extensively as a constituent of igneous rocks, quartz and sand.                 •
The silica content may serve to differentiate sediment due to                   |
erosion from that of industrial and municipal origin.

The classical gravimetric procedure is widely used for determining              I
silica in minerals and other materials e.g., portland cement, glass,
etc. which are high in silica content. (7.1, 7.1*-, 7»5«)

This method includes:                                                           •


                                                                                I
2.   Double evaporation - dehydration to isolate silica as
     its insoluble anhydride.                                                   •

3.   Ignition of the residue to constant weight.

k.   Volatilisation of silica as silicon tetrafluoride.                         |
                                                                                I
    Highly accurate results can be obtained using this method;
    however, it suffers the following disadvantages:  (a) it is                     •
    quite lengthy and (b) requires the use of expensive platinum                    •
    equipment.  This extreme accuracy is not required when
    monitoring the silica composition of a large number of sediments.
                                                                                I
    A modified gravimetric procedure is here proposed for the deter-
    mination of "uncorrected silica'(7»2) in sediments.  Pure or                     .
    corrected silica can be reported only after the residue has                     •
    been subjected to the hydrofluoric acid volatilization procedure.               *
    "Uncorrected silica" is hereafter referred to as "silica."
                                                                                I

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2.3  Watch glasses, to fit evaporating dishes.                                  •

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                                                                                 93
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Silica in Bottom Sediments


    2.5  Hot plates, large.


    2.6  Glass stirring rods.


    2.7  Filter paper.


    2.8  Filtering funnels and rack.


    2.9  Beakers, 400 ml capacity.


    2.10 Hot water wash bottles.


    2.11 Drying oven, 110°C.


    2.12 Fisher burners.


    2.13 Ring stand and rings.


    2.14 Clay triangles.


3.  Reagents


    3.1  Hydrochloric acid concentrated.


    3.2  Hydrochloric acid, 10$.


    3-3  Nitric acid, concentrated.


4.  Procedure


    Start with the residue from the determination of volatile solids,
    contained in a porcelain evaporating dish.


    4.1  Add 30 nil of concentrated HC1 to the residue.


    4.2  Pulverize with a pestle,  taking care not to lose any
         sample in the process.


    4.3  Wash the pestle with distilled water using a rubber
         policeman to dislodge adhering particles.


    4.4  Add 1 ml of concentrated HNO_ to each sample.


    4.5  Place the evaporating dish on a hot plate, cover with a
         watch glass and evaporate to dryness.


    4.6  Transfer the evaporating dish from the hot plate to a
         drying oven and bake at 110°C for 1 hour.

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Silica in Bottom Sediments
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    k.J  Set baked residue back on the hot plate, add 15 ml
         concentrated HC1 and heat to dissolve the difficult                        •
         soluble salts.                                                             |

    k.Q  Wash any adhering material off the watch glass into the                    •
         evaporating dish with about 25 ml of distilled water.                      •

    k.9  Continue heating until all soluble salts are dissolved,
         filter while hot through an ashless filter paper into                      I
         a kOO ml beaker (save this filtrate for subsequent                         I
         analysis).

    If-. 10 Wash any residue remaining in the evaporating dish onto                    Jj
         the filter paper using a rubber policeman to aid in the
         removal of adhering particles.                                             _

    U.ll Wash the residue retained on the filter paper free of                      •
         soluble salts with hot 10% HC1 (about 10 times).
                    x 100
        '2   g dry sample wt

6.  Discussion of Method
                                                                                    I
    U.12 Transfer the filter paper containing all the residue to
         a tared porcelain crucible.

    4.13 Dry and ignite to constant weight at the hottest                           •
         temperature attainable with a bunsen burner.

    k.lk Cool with cover on, weigh and calculate the % SiO.                         •

5.  Calculations

                                                                                    I
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    The modified gravimetric method for the determination of silica
    in sediments utilizes the residue from the determination of                     •
    volatile solids.  It is convenient to use this residue since                    •
    ignition at 600°C has destroyed interfering organic matter.
    Furthermore, ignition has served to prepare the sample for
    analysis,  by converting certain insoluble silicates to calcium                  I
    silicate,  which is acid soluble: (7.2)                                          •

         Equation (7»l)                                                             •
                      Si02 + CaCO  600°C    002 / CaSiO                             |


    This conversion is highly probable when sediments are ignited                   •
    since they contain substantial amounts of calcium carbonate.                    |
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                                                                    95

Silica in Bottom Sediments



    The sodium carbonate fusion serves a like function.

         Equation (2)
    Since ignition is noted to serve the same function as fusion,
    the latter step vas excluded from the modified procedure.


    The modified procedure also suggests that porcelain ware be used
    in lieu of platinum.  The literature noted that strong hydro-
    chloric and nitric acids, used to decompose the sample, might
    dissolve some silicates from the porcelain evaporating dish. (7»3)
    Hovever, it is further noted that, any silicates thus gained hope-
    fully will be negated by adsorption losses when the dehydrated
    silica is removed from the dish (silica being tenaciously adsorbed
    by porcelain) .


    The most time consuming step in the classical gravimetric silica
    procedure is the one which involves the double evaporation-
    dehydration of the sample.  A reference to the literature indicates
    that a single evaporation-dehydration step would recover 97-99$ of
    the total silica present in a sample. (7»3)


    Investigation has confirmed this recovery rate (See Table l).
    A single evaporation-dehydration is used in the modified procedure,
    since in surveillance work the relative change over a period of time
    rather than exact amount is most important.


    It is expedient to utilize the filtrate from the silica procedure
    for subsequent determinations of iron and manganese, since inter-
    fering organic matter and silica have been removed.  (See iron and
    manganese analysis ) .


    Finally, as noted in the General Discussion (Section l), silica as
    determined by the modified procedure is "uncorrected", since the
    residue was not subjected to a hydrofluoric acid volatilization.


7.  References


    7«i  Snell, F.D., & Biff en, P.M., Commercial Methods of Analysis,
         New York:  McGraw Hill, 19^, pp 176-191


    7«2  Ayres, G.H., Quantitative Chemical Analysis, New York:
         Harper Bros., 1958, 2^5-249.

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                                                                    96
Silica in Bottom Sediments
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     7.3     Kolthoff, I.M. & Sandell, E.B., Textbook of Quantitative
             Inorganic Analysis, New York:  MacMillan Co., 1946, p.402-403.         •
     7-4     Standard Methods for the Examination of Water and Wastewater,
             12th ed., APHA, Inc., N.Y., 1965 258-260.                              _
     7.5     "Methods for Collection & Analysis of Water Samples,"                  ™
             Geological Survey Water Supply Paper, 1454, I960, pp 259-263.
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1
1




1

1

1
VB
•
*
1
•
1



1

1

1
1
1
1
1
1



Sanple
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20





SILICA RECOVERY -
A
Wt. of Residue
Frca Single
Evap.-Eehy.
2.9732
3.2149
1.3853
1.3932
2.5408
1.0645
2.4757
1.9985
1.4854
3.5845
4.6403
1.0746
1.1986
5.1323
1.6429
2.1838
1.3859
3.7H4
1.5161
1.0811





TABIS 1
SINGLE VS DOUBLE EVAPORATION
B
Wt. of Residua
3?ro:a Second
Evap. -Bshy
0.0032
0.0022
0.0040
0.0015
0.0015
0.0008
0.0003
O.oo4o
0.0016
0.0110
o.oio4
0.0017
0.0037
0.0089
0.0063
0.0008
O.OOS4
0.0007
0.0021
0.0004






Total
Residue
A + B
2.9764
3.2171
1.3893
1.3947
2.5423
1.0653
2.4760
2.0025
1.4870
3.5955
4.6507
1.0763
1.2023
5.1412
1.6492
2.1846
1.3943
3.7121
1.5182
1.0815





97

$ Silica
Recovery By
Single Svap.
99.89
99.93
99.71
99.89
99.94
99.92
99.98
99.80
99.89
99.69
S9.77
99.84
99.69
99.82
99-61
99.96
99.39
99.98
99.86
99.96






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                                                                    98

                        MANGANESE IN BOTTOM SEDIMENTS
1.  General Discussion
                                                                                   I
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    1.1  Principle:  Manganese is readily determined in small concen-
         trations by oxidizing it in acidic solution to  the intensely             _
         colored permanganate ion and measuring the absorbance of the              •
         resulting solution at a wavelength of 525 n^i •                              ™


         After the sample has been digested with acid to oxidize                   |
         organic matter and to volatilize chlorides as  HC1,  phosphoric
         acid is added to form a colorless complex with ferric ion.                •
         Oxidation with persulfate ion is carried out in hot acidic                •
         solution in the presence of silver as a catalyst.  The color
         is stable for 2k hours.

    1.2  Interferences:  The most important interferences are organic              |
         matter, chloride ion and other reducing substances.  These are
         taken care of in the ashing steja.     Other colored ions,                 tm
         notably ferric iron, copper, nickel and chromate contribute to            •
         the color.  The interference of iron is overcome by the addition
         of phosphoric acid.  That of the other ions is compensated for
         using a portion of the sample from which the permanganate color           I
         has been bleached for a blank.                                            •

    1.3  Minimum detectable concentration:  If the volume of the sample
         is 100 ml, the minimum detectable concentration using a 1 inch
         cell is 0.01 mg/1.

2.  Apparatus

    Spectrophotometer equipped with absorption cells providing a light
    path of 1 inch or longer, for use at 525 EM.                                   I

3.  Reagents

    3.1  Stock manganese solution:  Dissolve 1.8g potassium permanganate           •
         in about ^50 ml distilled water in a liter flask and heat k-5
         hours at 70°-80t! protecting the mouth of the flask from dust.             _
         Within 2k hours standardize oxidimetrically against sodium                •
         oxalate.                                                                  ™

    3-2  Standard manganese solution:  The volume of stock solution                •
         required to prepare a liter of solution containing 50 mg/1 Mn             |
         is U.55 divided by the normality of KMnO^.  Transfer exactly
         this volume to a pyrex 1-liter volumetric flask.  Add 5 ml                M
         cone. HgSOlj. and then NaHSO- dropwise with stirring until the              •
         pink color disappears.  Boll gently for a few minutes to remove
         excess S02«  Cool and dilute to volume with distilled water.
         1 ml =  0.05 mg Mn.                                                       •
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                                                                     99
Manganese in Bottom Sediments                                        **
     3.3  Sulfuric acid, cone.

     3.U  Special reagent:  Dissolve 18.7500 g of mercuric sulfate in
          100 ml cone. HNO_ and 200 ml distilled water.  Add 200 ml
          85 percent phospnoric acid and 0.85 g silver nitrate and
          dilute the cooled solution to 1 liter.

     3.5  Ammonium perfiulfate, reagent grade.

     3.6  Sodium "bisulfite solution:  Dissolve lOg NaHSO^ in 90 ml
          distilled vater.
                  3.7  Sodium oxalate, primary standard, Na2C20^.  (0.2 —O.k g
                       anhydrous lCO^ weighed to nearest 0.1 ing.)
4.  Procedure


    4.1  Take an aliquot of the filtrate from the determination of
         silica containing not more than 2.0 mg of Mn.


    4.2  Transfer this aliquot to a 400 ml beaker, add 5.0 ml con-
         centrated I^SO^.  Cover loosely with a watch glass and
         evaporate on a hot plate to the copious evolution of SOo
         fumes.


    U. 3  Transfer the residue to a 250 ml Erlenmeyer flask, using a
         minimum amount of water, and again bring to fumes.


    4.4  Cool, add approximately 75 ml of distilled water and boil
         to dissolve the soluble salts.


    4.5  Allow to cool, add 20 ml of the special reagent and 1 g of
         ammonium persulf ate,  (Wty)2Sg)Q (use a 1 g Each measuring
         spoon for this addition).


    4.6  Quickly heat to boiling and continue to boil for 2 minutes.


    4.7  Remove the solution from the hot plate, add 0.2 g persulf ate
         (use a 0.2 g Hach measuring spoon) and allow to stand for
         1 minute before transferring to an ice bath for further cooling.


    4.8  Adjust the volume of solution in each flask to 100 ml with
         distilled water.

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                                                                     100



Manganese in Bottom Sediments                                                      I




    U.9  Read the absorbance of each solution versus a blank in a                  •

         spectrophotometer set at a wavelength of 525 ^•                          I



    IK 10 Prepare calibration curve covering the range 0-5 mg/1.



5.  Calculation                                                                    ™
    5.1  Wet Basis                                                                V



                                       -  x 1000


                            in aliquot)
            A     mg Std.   OP Sample        .,.-.-

         »*/** = ODStdT X (grams Sample  X 10°°
               " ^Solids(decimal fraction)



6.  Reference
    5.2  Dry Basis

                 ing/kg wet basis	                                      I
               " <& Solids  (decimal fraction)                                      I



    Standard Methods for the Examination of Water and Wastewater,
    12th ed., APHA, Inc., N.Y., 1965, l»89-492.                                     .




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                                                                101
             OXIDATION-REDUCTION POTENTIAL MEASUREMENTS



The oxidation-reduction (redox or Eh) potential is defined as the
electromotive force developed by a platinum electrode immersed in the
vater or bottom sediment, referred to the standard hydrogen electrode.

This is a measure of the oxygen potential of the bottom sediments.
When % is positive, this indicates the presence of oxygen or the material
is in the oxidized state and when it is negative the absence of oxygen is
indicated or the material is in the reduced state.               An Eh
measurement would give a good indication whether the iron was in the ferric
state or the ferrous state.

To measure the potential, a Beckman Zeromatic pH Meter or a Beckman
Model N is used in connection with a calomel (saturated KCL solution
used) and platinum electrode system.  Instructions are followed for the
specific meter to obtain MV readings.  Eh is measured by inserting the
electrodes just into the surface of the undisturbed sediment (about 1/2").

The oxidation-reduction potential of the sample in millivolts referred
to the hydrogen scale is calculated as follows:

Oxidation-reduction potential, Mv s E - C where:

        E = electromotive force, in millivolts of the cell

        C = potential, in millivolts of the saturated calomel
            electrode referred to the hydrogen scale.

Note:  There should not be any electrical interferences in the immediate
       locale.

Reference

    Manual on Industrial Water, Second Ed., 1966 Printing,
    No. 3A8-1, pages 578-582.

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 .3. Environmental Protection Agency
Region 5, Library ,'PL-1 /• J)                                                                 I
           «-                       c*                                                    •
        ,
77 West Jac«-.*-n Poulard. 12th Hoc*
        IL  er-;r.j v/»
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