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
Environmental
Protection
Agency
EPA 335
Official Business
Penalty for Private Use $300
U S
oooo30604
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