United States
Environmental Protection
Agency
Environmental Monitoring and Support
Laboratory
Cincinnati OH 45268
Research and Development
EPA-600/S4-83-002  Apr. 1983
Project Summary
Development  and  Evaluation  of
Analytical  Test  Procedures  for
Priority  Pollutants
C. S. Monteith and Daniel F. Bender
  Analytical methods were developed
for the determination of cyanide and of
total phenolic compounds in solid/
semisolid samples of environmental
importance. The approach to the prob-
lem was to: 1) select the most prom-
ising isolation and quantification meth-
ods from a literature search; 2) empiri-
cally evaluate them in order to choose
the methods for further study, 3) modify
and optimize the procedural steps of
the methods; and 4) validate the meth-
ods by applying them to a variety of
actual solid/semisolid samples.  Each
method consisted of two steps: isola-
tion of the analyte and quantification
of the analyte. Isolation studies were
performed using radiolabeled cyanide
and radiolabeled phenolic compounds
to spike representative matrices.
  The method which was selected for
the determination of cyanide in solid/
semisolid matrices involves distillation
of cyanide from an acidified slurry of
the sample, followed by quantification
using the pyridine-barbituric acid pro-
cedure. The method which was selec-
ted for the determination of total
phenolic compounds involves extrac-
tion of an acidified slurry of sample
with methylene chloride. The methylene
chloride is then extracted with a sodium
hydroxide solution and quantified using
the 4-aminoantipyrene (4-AAP)  pro-
cedure.
  This Project Summary was developed
by EPA's Environmental Monitoring
and Support Laboratory, Cincinnati.
OH to announce key findings of the
research project that is fully docu-
mented in a separate report of the
same title (see Project Report ordering
information at back).

Introduction
  Cyanide complexes are found in the
Wastes of many industrial processes such
as electroplating, metal refining, and steel
hardening.   They  are intermediates in
polymer synthesis and are used as fumi-
gants in produce storage. Phenolic wastes
are generated by the petroleum, plastics,
and organic chemical industries.   Both
compounds may accumulate in sediments
and are taken up by aquatic plants and
animals in a variety of ways.
  Under the Federal Water Pollution Con-
trol  Act Amendments of  1972 (Public
Law  92-500), proposed discharges of
dredged  or fill material into navigable
waters will be evaluated  by the  U.  S.
Environmental Protection Agency.  The
procedures developed in this study will be
considered as potential  avenues for ob-
taining analytical information  concerning
the material in question.
Results

Cyanide
  For the isolation of cyanide from the
sample, distillation was clearly the most
viable alternative.  Microdiffusion, which
has been used for biological samples for
volatile cyanides,  is  severely limited by
sample size requirements. Solvent extrac-
tion of cyanide from a solution of sodium
hydroxide and ammonium hydroxide with
hexane and with methylene chloride gave
less than 1 % recovery and was therefore
eliminated from further consideration.

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   Hydrogen  cyanide is  distilled under
 aeration from an acidified slurry contain-
 ing magnesium chloride.  The gas passes
 through an absorber containing lead ace-
 tate solution in order to remove interfering
 sulfur compounds; then it passes into an
 absorber  containing  sodium hydroxide
 solution for  final collection.   For thick
 slurries resulting from high sample volume
 and weight simultaneous stirring with  a
 magnetic  stirrer prevents bumping.
  Radiolabeled cyanide was spiked onto
several matrices which were then distilled.
The distillation efficiency was followed by
withdrawing aliquots for scintillation count-
ing. The recovery of cyanide from spiked
solid/semisolid samples varied  with the
nature of the sample matrix and the distil-
lation time. Typically, fish tissue yielded
40% of the spike, lake bottom sediment
yielded 63-8696 of the spike, and sludge
yielded 43-52% of the spike using a two-
hour distillation period.
  Seven methods for the quantification of
cyanide were experimentally evaluated.
Five of  these were  spectrophotometric
methods involving the use of: 1) pyridine-
barbituric acid reagent; 2) tris( 1,10-phen-
anthroline) iron (II) reagent 3) mercuric
chloranilate reagent  4) ammoniacal nickel
chloride reagent  and 5)  /yyV-dimethyl-
formamide reagent  The other two were:
1) an ion selective electrode method; and
2) a gas chromatographic method involv-
ing the determination cyanogen bromide
after bromination of the sample.
  Though the gas chromatograph is the
most sensitive instrument for these anal-
yses,  the  necessary preliminary solvent
extraction produced an  average of only
50%  recovery.   The  pyridine-barbituric
acid procedure was  the next most sensi-
tive and produced an average 97% recovery
of spiked distilled water samples.
  Experiments  which  were performed
using spiked fish  tissue  demonstrated
both the necessity of the lead acetate trap
and the near quantitative recovery of
spikes.
  The validation study involved recover-
 ing spikes at the 2.6 jug/g level and the
20 jLig/g  level from ten  solid/semisolid
 matrices.  The matrices and the average
 percent recovery of the spikes  are: lake
 bottom sediment (84 ± 6%); industrial
sludge (84 ± 12%); industrial solid waste
(94 ± 11 %); industrial processing slag
(94 ± 3%); incinerator fly ash (95 ± 7%);
contaminated soil  (103 ± 12%);  fish
tissue (94 ± 6%); microinvertebrates
(shrimp) 92 ± 6%);  algae (76 ± 1 %); and
vegetation (84 ± 10%).
Phenol
  For the isolation of phenolic compou nds
from the sample, solvent extraction with
methylene chloride was selected as the
most successful alternative. Steam distilla-
tion, which is the recommended method
for aqueous samples, gave near quantita-
tive recovery of phenol,  but  almost no
recovery of para- nitrophenol and of 2,4,5-
trichlorophenol. While they are potential
separation methods, liquid chromatography
and carbon adsorption techniques are of
limited capacity and require further study.
Soxhlet extraction with methylene chloride
appears to be almost as  efficient as ex-
traction with stirring only.  Further study of
this alternative may produce improvement
  The acidified and blended sample slurry
is extracted with methylene chloride which
in turn is extracted with  diluted sodium
hydroxide solution for the quantification
step.

  Radiolabeled phenolic compounds were
spiked onto several  matrices and  were
then extracted.  The extraction efficiency
was determined by scintillation counting
of aliquots of the methylene chloride. After
two hours of stirring, the methylene chlo-
ride yielded from 72-98% of three radio-
labeled phenolic compounds which were
spiked onto fish tissue, lake sediment and
sludge.
  Three methods for the quantification of
total phenolic compounds were evaluated
by performing seven  determinations on
spiked aqueous samples.   The 4AAP
method, the 3-methyl-2-benzothiazolinone
hydrazone colorimetric method (MBTH),
and the  bathochromic shift  procedure
(two variations  - one  using a laboratory
spectrophotometer and the other using a
commercial ultraviolet ratio spectrophoto-
meter) were evaluated.  The  sensitivity,
range, precision, and accuracy were meas-
ured.   The commercial ultraviolet ratio
spectrophotometer was the most sensi-
tive and the laboratory spectrophotometer
was least sensitive.
The4-AAP method was selected because:

  1. It has been shown to work on a wide
    variety of environmentally important
    aqueous matrices.
  2. The results can  be compared to an
    abundance of accumulated data
  3. Phenolic mixtures determined by the
    two methods would most likely not
     be linearly related since the substi-
    tuent effects  are not quantitatively
     related.
  4. The ultraviolet  ratio spectrophoto-
     meter is not available in most labora-
    tories where analyses on solid/semi-
     solid matrices wilf be performed.
  5. The sensitivity of the 4AAP method
     is already more than adequate, thus
     the advantage of the ultraviolet ratio
     spectrophotometer is minimal.

  The validation study involved recover-
ing spikes at the 0.80 jug/g level and the
4.0 /ig/g  level  from  ten solid/semisolid'
matrices.  The matrices and the average
percent recovery of the  spikes  are: lake
bottom  sediment (75 ±  5%); industrial
sludge (55 ± 4%); industrial solid waste
(71 ±  13%); industrial  processing slag
(36  ±  10%);  fish  tissue  (26 ±  3%);
microinvertebrates (shrimp)  (39 ± 5%);
algae (34 ± 10%); and vegetation (42 ±
9%).

Preservation
  A concurrent preservation study was
performed.  For cyanide, three  methods
were studied: 1) storage at less than 0°C;
2) storage at 4-6° C; and 3) storage at 4-6° C
with sodium hydroxide  addition.    For
phenol,  three methods were studied: 1)
storage at less than 0°C; 2) storage at 4-
6°C; and 3) storage at 4-6°C with  phos-
phoric acid and copper sulfate addition.
  The best methods for preserving cya-
nide are freezing in dry ice and storage at
4-6°C after the addition of sodium  hy-
droxide In this study, the former method
was used in order to avoid the difficulties
encountered when using sodium hydroxide
solution in the field.
   Freezing in  dry ice was  clearly  the
superior method for  preserving phenolic
compounds.  The other alternatives for
preserving phenolics allowed extremely
rapid deterioration.  For verification, cya-
nide-containing  sludge from a  chemical
processing manufacturer and phenol-con-
taining soil which had been contaminated
by a chemical manufacturing plant were
stored at less than 0°C for up to 16 days.
Periodic analyses by the selected colori-
metric  procedures showed virtually no
loss of analyte.
  This project summary was prepared by
Daniel F. Bender of the U. S. Environmental
Protection  Agency from  the report pre-
pared by  Carolyn  Monteith of the Gulf
South Research Institute. Daniel F. Bender
is the U.S. EPA Project Officer.  Carolyn
Monteith is the GSRI Project Manager.

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     C. S. Monteithis with Gulf South Research Institute. New Orleans, LA 70186 and
       the EPA author Daniel F. Bender (also the EPA Project Officer, see below) is
       with the Environmental Monitoring and Support Laboratory, Cincinnati, OH
       45268.
     The complete report, entitled "Development and Evaluation of Analytical Test
       Procedures for Priority Pollutants," f Order No. PB 83-166 181; Cost: $13.00,
       subject to change) will be available only from:
             National Technical Information Service
             5285 Port Royal Road
             Springfield, VA 22161
             Telephone: 703-487-4650
     The EPA Project Officer can be contacted at:
             Environmental Monitoring and Support Laboratory
             U.S. Environmental Protection Agency
             Cincinnati, OH 45268
                    OU.S. Government Printing Office:  1983-655-017/7030
United States
Environmental Protection
Agency
    Center for Environmental Research
    Information
    Cincinnati OH 45268
Postage and
Fees Paid
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Protection
Agency
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Penalty for Private Use $300
U  S
                                     oooo30604

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