EPA-600/4-76-048
September 1976
Environmental Monitoring Series
              EVALUATION OF  INSTRUMENT  FOR  THE
              DETERMINATION  OF  PHENOL IN  WATER
                                Environmental Monitoring and Support Laboratory
                                       Office of Research and Development
                                      U.S. Environmental Protection Agency
                                              Cincinnati. Ohio 45268

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                RESEARCH REPORTING SERIES

Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency,  have been  grouped into  five series. These five  broad
categories were established to facilitate further development and application of
environmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The five series are:

     1.    Environmental Health Effects Research
     2.    Environmental Protection Technology
     3.    Ecological Research
     4.    Environmental Monitoring
     5.    Socioeconomic Environmental Studies

This report has been assigned to the ENVIRONMENTAL MONITORING series.
This series describes research conducted to develop new or improved methods
and  instrumentation for the identification and  quantification of environmental
pollutants at the lowest conceivably significant concentrations. It also includes
studies to determine the ambient concentrations of pollutants in the environment
and/or the variance of pollutants as a function of time or meteorological factors.
 This document is available to the public through the National Technical Informa-
 tion Service. Springfield, Virginia 22161.

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                                     EPA-600/4-76-048
                                     September 1976
       EVALUATION OF INSTRUMENT FOR THE

       DETERMINATION OF PHENOL IN WATER
                      By

               L. Shelbert Smith
            Department of Chemistry
           Central State University
           Wilberforce, Ohio  45384
             Grant No. R803172-01
                Project Officer

             Morris E.  Gales, Jr.
        Environmental Protection Agency
Environmental Monitoring and Support Laboratory
            Cincinnati, Ohio  45268
ENVIRONMENTAL MONITORING AND SUPPORT LABORATORY
      OFFICE OF RESEARCH AND DEVELOPMENT
     U.S.  ENVIRONMENTAL PROTECTION AGENCY
            CINCINNATI, OHIO  45268

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                                DISCLAIMER

     This report has been reviewed by the Environmental  Monitoring and
Support Laboratory, U.S.  Environmental Protection Agency,  and approved
for publication.  Approval does not signify that the contents neces-
sarily reflect the views  and policies of the U.  S. Environmental  Pro-
tection Agency, nor does  mention of trade names  or commercial products
constitute endorsement or recommendation for use.
                                    11

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                                 FOREWORD

     Environmental measurements are required to determine the quality of
ambient waters and the character of waste effluents.  The Environmental
Monitoring and Support Laboratory-Cincinnati conducts research to:

     o  Develop and evaluate techniques to measure the presence
        and concentration of physical, chemical, and radiological
        pollutants in water, wastewater, bottom sediments, and
        solid waste.

     o  Investigate methods for the concentration, recovery, and
        identification of viruses, bacteria and other microbiological
        organisms in water.  Conduct studies to determine the response
        of aquatic organisms to water quality.

     o  Conduct an Agency-wide quality assurance program to assure
        standardization and quality control of systems for monitoring
        water and wastewater.

     There is an ever-increasing interest in the use of instrumental
methods to analyze water and waste samples, whether the resulting data
are to be used for research, surveillance, compliance monitoring, or
enforcement purposes.   Accordingly, the Environmental Monitoring and
Support Laboratory has an on-going methods research effort in the
development, evaluation, and modification of instrumental methods.
This particular report pertains to the evaluation of the PH-2 Phenol
Detector.  The method has potential routine application for the analysis
of phenols in surface waters and domestic and industrial wastes.
                                   Dwight G.  Ballinger,  Director
                          Environmental Monitoring § Support Laboratory
                                            Cincinnati
                                   111

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                                 ABSTRACT

     This report presents the evaluation of a commercially available
ultraviolet spectrophotometer designed specifically for the deter-
mination of phenolic compounds in water and wastewater.  The analytical
procedure is based on the measurement of the bathochromic shift that
occurs with phenol when the pH of the solution is changed from pH 4
to pH 12.  The method allows for the rapid, sensitive and reproducible
analysis of phenols.  The time of a single analysis, exclusive of any
distillation that may be necessary, is less than five minutes.  The
results show that the determination of phenols in water can be obtained
with an accuracy of ±5% in the range of fifty parts per billion to
fifty parts per million and with an accuracy of ±12% in the range of
five parts per billion to fifty parts per billion.  The results of
this study were based on the analysis of a variety of synthetic samples
of phenol and of substituted phenols and their mixtures, and of real
samples from a number of sampling sites in the local area.
                                    IV

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                             CONTENTS

Foreword	   iii

Abstract	     iv

Figures	     vi

Tables	   vii

Acknowledgement	viii

   1.   Introduction  	     1
   2.   Conclusions 	     5
   3.   Recommendations 	     7
   4.   Objectives	     8
   5.   Instrument Design 	     9
   6.   Apparatus and Reagents	     10
   7.   Experimental	     11
   8.   Discussion	     22

References	     26

Appendices	     29

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                              FIGURES



Number                                                      Page



   1   4-AAP Method (CHCl- Extraction)                       13
                         O


   2   4-AAP Method (Direct Photometric)                     14



   3   PH-2 Instrument                                       17
                                 VI

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                              TABLES

Number                                                       Page

   I   Phenolic Concentration Values Determined by the
       4-AAP Method	   12

  II   4-AAP Method -  Preparation of Working Curve	   12

 III   Data for Determination of Calibration Constant,
       PH-2 Instrument	   16

  IV   Comparison of Methods with Substituted Phenols ...   18

   V   Comparison of Results Obtained for Phenols in
       Distilled Water by the 4-AAP and the PH-2 Methods.  .   21

  VI   Recovery of Phenol by the PH-2 Instrument Method .  .   22

 VII   Phenol Concentration of Industrial Samples
       (Average Values)  	   23
                                VII

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                             ACKNOWLEDGEMENTS

     Much of the collection and analyses of the data of this project
were performed by Ms.  Helen Addie and Ms. Debra Davis, senior students
in the Chemistry Department at Central State University.  Their dedi-
cated and careful work and technical assistance are hereby acknowledged
with sincere thanks.

     The assistance and encouragement of Mr. Joseph Suhre, Chemist,  Miami
Conseryancy District,  Dayton, Ohio, who provided us with many of the
real samples, is hereby acknowledged with sincere thanks.

     Dr. John E. Barker and Mr. Harry E. Clinton, Environmental Engineering
Department, Armco Steel Company, Middletown, Ohio furnished samples  of
process waters for this project.  Their assistance is hereby acknowledged.

     Mr. Donald Cunningham, Water Department, Central State University,
furnished samples of process waters for this project and his assistance
is hereby acknowledged.

     Mr. John D. Johnson, Mr. James W. Cutler and Mr. Fred L. Corcoran
of Spectro Products, Inc. provided advice and ready assistance in tech-
nical problems during the course of this work.

     The continuous assistance, encouragement and advice of Mr. Morris
Gales, Chemist, U. S.  Environmental Protection Agency, Cincinnati, Ohio,
during the course of this project is gratefully acknowledged.  It was with
his continuous support that this project was initiated and completed.
                                   Vlll

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                              INTRODUCTION

     For many years little initiative was taken to abate the pollution of
our environment and the misuse of our natural resources.  In fact, com-
munities took the view that the benefits accruing to them from the
industries that were polluting the environment outweighed the backlash
of pollution environmental degradation.  Thus, most communities and
governmental agencies were insensitive to the problem and regarded pol-
lution as an unfortunate, but nonetheless acceptable, side effect of
industrialization.  However, within the past few years, the public has
developed a greater cause for concern regarding pollution control and
management of the environment.  The concern has developed because of the
greater public awareness about the growing environmental deterioration
and the need to maintain acceptable environmental standards for health-
ful living of a growing population.  As the awareness of the pollution
problem has sharpened, it has become clear that people and other sources
have been as guilty of pollution of the environment as industries (1,2).
However, it is obvious that much of the environmental deterioration is
a consequence, directly and indirectly: of the technological advances
that have occurred (1).  Both population growth and the demand for more
goods and services have contributed to the rapid increase in industriali-
zation and technological advances.  Thus, the advances in technology and
industrialization, with a greater demand of our physical and human
resources, have added new dimensions to the problems of controlling
further environmental deterioration.  Because of public concern,  the
government now has taken steps to more adequately manage and control
our environment by establishing guidelines and regulations to monitor
and control pollution and the misuse of our natural resources.   The
success of these steps depends in a large measure on identifying serious
pollutants, measuring their quantities, and developing methods to remove
them so that they do not enter the environment (3).

     The deterioration of the environment is well illustrated by the
exploitation and misuse of our water resources (1,4-6).   A cursory exam-
ination of many of our water sources clearly demonstrates that they have
become more polluted over the years.  However, water pollution is of
specific concern to everyone for it is our rivers, streams,  and lakes
that are a major source of food, that are widely used for recreational
purposes, and, more importantly, that are sources of drinking water for
many of our communities.   At the same time, it has been noted that the
natural processes which help restore water to its natural condition are
hampered by the increase of pollutants that find their way into the
streams, lakes, and rivers.   While much of the pollution of our water
resources has been attributed to industrialization and urbanization,  it
is predictable that both of these factors will continue to increase in
the future.  Therefore, better water management is necessary to maintain,
protect and upgrade water quality for present and future potential uses
by an increasing population and industrialization.

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     One of the major concerns of the environmental scientist has been
the marked increase in the number of organic compounds that find their
way into our water sources.  Many of these organic pollutants are derived
from industrial plants, but agricultural run-off, mine drainage, and
domestic wastes contribute a significant share of responsibility.  Of the
large volume of organic chemicals produced by industry, phenol and phenol
derivatives widely used in the manufacture of dyes, polymers, pesticides
and pharmaceuticals represent a significant percentage of the organics
that find their way into the waste products dumped into our rivers, lakes
and streams.  These phenols are troublesome in that they are not removed
completely in waste treatment plants and thus become the source of offen-
sive odor and taste in drinking water (7-9).  Many phenolic compounds
found in water are chlorinated during the process of water treatment and
become the source of the objectionable odor and taste in water  (10).
Significantly, for example, it has been reported that chlorphenol produces
an unpleasant taste in fish at concentrations of 0.1 microgram per liter
(yg/1) (11)-  Although research has not shown that these phenolic com-
pounds are harmful or a hazard to health at very low concentrations, the
odor and taste that they impart to drinking water, even at very low levels,
make them serious water pollutants.  Thus, the need to monitor and control
the amount of phenolic compounds and other organics in water and wastewater
is of concern to the environmental scientist.

     Analytical chemistry  and instrumentation have long played a vital role
in the detection and identification of the many pollutants in water.  As
the number of identifiable pollutants increases and their toxicities are
determined, it is necessary that improved techniques be developed to
measure the pollutants at  much lower levels than available by using present
methods.  Such analytical  methods must be applicable to a variety of water
samples and in the presence of a variety of contaminants.  Much effort has
been undertaken, therefore, in analytical chemistry to improve on the pro-
cedures for the analysis of organics, such as phenolic compounds.

     A review of the literature reveals that a number of procedures for
the determination of phenolic compounds in water have been studied.  It
should be noted that,  to date, all of the methods depend on the conden-
sation or reaction of  some reagent with the aromatic ring of phenol and
not with the hydroxyl  function of phenol.  Unfortunately, many of these
procedures have a variety  of shortcomings.  Thus, there is still a need
for a rapid, reliable  procedure for the determination of phenolic com-
pounds that is applicable  to a wide range of concentrations and types.

     One of the early  developed and frequently used method for  the deter-
mination of phenolic substances was that proposed by Gibbs (12).  This
analytical procedure is based on the reaction of phenolic compounds with
dibromoquinone in a buffered alkaline solution to form dibromoindephenol
dyes.

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The dye is measured colorimetrically at 610-630 nm, and the concentration
is determined from a prepared standard curve.  Concentrations of phenolic
compounds in the range of 0 to 10 parts per million (ppm) can be measured.
In the concentration range of 3 to 100 parts per billion (ppb), the dye
must be concentrated by extraction with n-butyl alcohol and measured at
670 nm.  In this procedure, it has been found that control of pH is criti-
cal, because any variation affects color development,  as do temperature
changes.  It should be noted that the procedure requires from 6 to 24 hours
for an analysis, because various phenolic compounds react at different
rates.  Even so, some phenolic compounds, such as p-cresol, do not react
in this method.  Studies of the technique indicate that the lower concen-
tration limit of phenolic compounds that can be determined is about 50
ppb, and the error in measurement is about 2-3%.  However, the problems
associated with this procedure have led to a study of  others (13-16).

     Another method that has been examined is the reaction of phenolic
compounds with nitrous acid to form a nitrosophenol.
                    HOMO
* H
The nitrous compound is then rearranged in an alkaline medium to form a
colored quinoid salt, which is measured colorimetrically at 420 nm (16-18).
In this procedure, certain amines and inorganic salts interfere somewhat
with the development of the color,  and up to 24 hours may be required for
the analysis.  Very few non-phenolic compounds interfere, but it has been
found that para-substituted phenolic compounds do react.   For low concen-
tration determinations, an extraction procedure is needed to concentrate
the phenolic compounds.  Again, this procedure has not been widely adopted
because of the time requirement and the consistently low results obtained.
However, precision of ±1% has been reported for this method.

     Recently, gas chromatography and infrared methods of analysis have
been developed for determination of phenolic compounds (16, 19-22).   Al-
though  these methods utilize instrumental procedures found in most lab-
oratories, the cost of the equipment can be prohibitive for many labor-
atories.  In gas chromatography, water samples require the extraction of
the organic compounds before the analysis.  Thus, considerable time may
be consumed in concentrating the organics from the water samples.   However,
gas chromatography can give reliable data on all types of phenolic com-
pounds, and this procedure is used primarily for the separation and
identification of phenolic compounds in the parts-per-mi11ion range.

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     The infrared absorption procedure is based on the ease of bromination
of phenolic compounds and the measurement of the shift of the 0-H infrared
band of the carbon tetrachloride (CC14.) extracted brominated compound.
The shift of the 0-H band occurs as a result of hydrogen bonding between
the 0-H group and the ortho bromine atom.  The magnitude of this shift
remains constant as long as the bromine substitutes ortho to the 0-H
group, which means if the di-ortho position is substituted with groups
other than bromine, phenol cannot be determined with this method.  However,
it has been found that this procedure is quite sensitive even in the
parts-per-billion range.  The limitations of this procedure appear to be
the cost of the equipment and that phenol cannot be determined,  if steric
effects keep them from brominating.

     The most commonly used procedure is that developed by Emerson (23-26),
which is based on the reaction of phenolic compounds with 4-aminoantipyrine
(4-AAP) in an alkaline oxidizing medium to form antipyrine dyes.
                                              «o

                                      CHj-CrzrC-M::^

The aqueous solution of the dye is measured colorimetrically at 510 nm,
and the concentration of phenolic compounds is determined as phenol by
comparison with a standard curve.  For concentrations in the range of
parts per billion, the dye is extracted with chloroform and its absorbance
measured at 460 nm.  This method has been used routinely in a variety of
applications for many years and has been adopted for the determination of
phenol by the American Society for Testing and Materials (27) and the
American Public Health Association (28).  Because of its acceptance as
the standard method, it has been examined closely for the effects that
variables have on the determination of phenolic compounds.  One of the
serious disadvantages of this procedure is that phenols having sub-
stituents in the para position do not react to give the colored dye.
However, it has been found that in the reaction, groups such as halogen,
hydroxyl, carboxyl, methoxy, and sulfonic acid are replaced and thus
the para substituted phenols react with the 4-AAP.  Careful studies have
been made of the effect that pH, order of addition, concentration of
reactants, temperature, and aging have on the development of the colored
dye and on the sensitivity of the determination (16, 29-34).  These studies
have led to the establishment of a widely used standard procedure which
give reproducible and reliable results in the useful range of 2.0 ppm to
20 ppb at an accuracy of ±2%.  Frequently, because of turbidity, samples
must be distilled before using the method, which adds to the time neces-
sary to obtain an analysis.  The time required is, nevertheless, much
less than that needed in the methods discussed above; hence it has
become the most widely used procedure for a wide variety of analyses.

     A recent modification and improvement of the 4-AAP method has been
developed by Friestad (35).  This method depends on the oxidative coupling

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of phenolic compounds with 3-methyl-2-benzothiazoline hydrazone CMBTH).
This method is not as dependent on the availability of the para position
in phenols, as the 4-AAP method is, since, if the para position is blocked,
the reaction will take place at a free ortho position.  The colored com-
pound obtained is measured spectrophotometrically at 520 nm and compared
with standard solutions.  As with the methods described above, this pro-
cedure determines all phenolic compounds as phenol.  Better results are
obtained by this method than those of the 4-AAP method, and the detection
limit is about 5 ppb.  Samples of the dye at very low phenol levels are
concentrated by extraction with chloroform.  Although this method extends
the range of phenolic compounds that can be detected and is less subject
to interferences than the 4-AAP method, analysis requires about one-
half hour.

     Within recent years, a sensitive ultraviolet spectrophotometric
method has been developed.  This method, first introduced by Murray (36)
for the analysis of phenolic compounds in gasoline fractions, makes use
of the bathochromic shift in the spectrum due to the formation of the
phenolate ion in alkaline solutions.  This method has been used for a
variety of phenolic compounds in many different applications, is the most
sensitive procedure developed, and does not suffer from many of the
problems associated with the other methods.  The procedure requires less
than 5 minutes for an analysis and is not affected by the structural
characteristics of the substituted phenols.  Other workers have refined
the procedure, and indications are that, in addition to greater sensi-
tivity and freedom from interference, the method is capable of detecting
phenols in much a lower concentration range than other procedures can
(16, 37-44).   The method obviously overcomes the difficult problems
encountered in the analysis of water and wastewater containing a variety
of phenolic compounds in varying concentrations.  The procedure has led
to the development of instrumentation for the specific analysis of
phenolic compounds in water and wastewater.  The objective of this
project was to evaluate this commercial instrument.

                               CONCLUSIONS

     A careful review of the literature shows clearly the limitations and
problems that have evolved in the development of analytical methods for
the determination of phenol in water and wastewater.   All of the proposed
methods depend on a reaction with the aromatic ring of phenol.  This fact
places limitations on the analysis of a variety of substituted phenols,
particularly ortho and para substituted phenols, since the 0-H group
directs reactions to these positions.  In addition to this problem, the
rate at which various test reagents react with the phenol ring varies
considerably.  The ultraviolet spectral method on which this work is

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based is dependent on the ability of the phenol 0-H group to ionize.  This
ionization, which occurs as the pH is shifted from acidic to basic con-
ditions, allows for correlation of the concentration and ion formation
of phenol.  The bathochromic shift which occurs as the acidity and basicity
are changed permits a reliable measure of phenol concentration.  Thus,
this method is dependent on the presence of the phenolic 0-H group and
independent of the positional substituents present.  However, it is rec-
ognized that the type of substituents can affect the absorptivity and the
bathochromic shift of some phenolic compounds, but the effect is minimal
in most instances.

     The PH-2 instrument evaluated provided several advantages over the
4-AAP method presently accepted as the standard procedure.  The PH-2 pro-
cedure requires, as the only reagents, one drop of concentrated acid and
two drops of saturated solution of alkali.  In the actual measurements,
one drop of concentrated phosphoric acid was sufficient to bring the phenol
solutions to a pH of 4.0, and two drops of saturated solution of sodium
hydroxide shifted the pH to a value of 12.  The procedure allows for a
rapid analysis since it involves setting the meter at zero with the solu-
tion at pH 4 and then reading the meter directly when base is added to
the solution to bring the pH to a value of 12.  It was found that the pH
was not critical and that approximate minimum pH values sufficed.  A
single measurement can be made in less than three minutes, exclusive of
any distillation that may be necessary.  However, more care and time are
needed if the expanded scale for ppb concentrations is used, because of
the extreme sensitivity of the dial in establishing the zero reading.
Nonetheless, no more than five minutes are needed, including dilution of
samples, for the determination and calculation of the results obtained.
It should also be noted that no more than a 50-ml sample is needed for a
single analysis, about 10-ml for use with the 2-cm cell or about 35-ml
for the 10-cm cell.

     As indicated by the experimental data, this procedure has the signifi-
cant advantage over the 4-AAP method in that it will detect para sub-
stituted phenols.  It should be noted that this ultraviolet spectral method
gives values which are higher than those derived from the 4-AAP methods.
This fact is also noted in the analysis of the industrial samples.  These
higher values probably are due to the fact that, in some instances, the
color formation reaction with the 4-AAP is incomplete which gives lower
values in this method while in the ultraviolet spectral method, the con-
version of the enol  (phenol) to the phenolate ion is essentially complete
and independent of the substituents.  Thus, this commercial instrument is
capable of providing a more complete assessment of the concentration..

     The disadvantage of the ultraviolet method is the problem of detect-
ing phenols which can exist as tautomers.  The experimental data show
that these compounds lie outside the absorption maxima for phenol and
the phenolate ion.  The existence of these substituted phenols as tauto-
mers is well documented in the literature and their observed spectral
behavior is not unexpected.  However, an appropriate procedure for the
determination of these substances should be established.

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     A representative and appropriate number of real  (industrial) samples
were also analyzed.  It is significant that a. wide range of concentrations
are represented and that, in general, the analyses are higher than with
the 4-AAP method.  Although for some samples, 4-AAP analyses were deter-
mined by the laboratories, the results were not available for this work.
These industrial samples represented a wide variety of sources, including
both rural and urban water treatment plants, industrial plants, and
foundries.  All of the samples were preserved with copper sulfate and
phosphoric acid, and it was necessary to distill each one prior to the
analysis.

     The error analysis of the data obtained from the several runs pro-
vides an indication of the accuracy and precision of the instrumental pro-
cedure.  The accuracy was determined by a measure of the observed and true
values for a series of solutions using the 2-cm cell and the 10-cm cell.
The results show a range of 1.5-7.0% accuracy for the 2-cm cell and
10-25% for the 10-cm cell.

     An important consequence of this work was that it provided an excel-
lent opportunity for the students who assisted in the project to apply
their theoretical training and knowledge to a very practical problem.  It
provided the students with an opportunity to extend their training in a
productive and challenging manner.  Thus, for those associated with the
project, the problem served a twofold purpose—the challenge of completing
a practical problem as well as applying theoretical training to a
commercial problem.

                             RECOMMENDATIONS

     The instrument provides for the measurement of phenolic compounds in
the range of 50 ppb to 50 ppm.  An expanded scale,  which increases the
sensitivity of the instrument, is provided for use in the low parts per
billion range.   Because of the increased sensitivity when used with the
expanded scale, the instrument requires more care (almost complete damping)
in setting the zero point.  Although the additional care and time involved
is not excessive,  it is recommended that a vernier dial be incorporated in
the instrument for use with the expanded scale.   This will permit a more
precise setting of the zero point with a less sensitive dial,  and thus
conserve time.

     The quantitative measurement of phenolic compounds in water and
wastewater by the PH-2 instrument is based on the bathochromic shift that
occurs in phenol when the pH of the solution is  shifted from acidic
(pH 4.0) to basic (pH 12.0)  conditions.   The instrument,  which uses hollow
cathode lamps as its light source, is designed to measure phenolic sub-
stances as phenol.   However,  it is obvious that  all phenolic substances
do not have the same absorption maxima as phenol.   This work shows that
certain substituted phenols give values which are considerably less  than
the theoretical values.   This has been ascribed  to the fact that these
phenolic substances have absorption maxima which lie outside the range
of the instrument.   It is recommended that consideration be given to

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adding another lamp to measure the bathochromic shift of the phenolic
substances which are incompletely detected.  In particular, this lamp
should be designed for the measurement of the chlorinated phenols which
are serious pollutants and which appear to be positively, but incompletely,
detected by the PH-2 instrument.  The lamp should be installed so that it
may be conveniently moved in and out of the measurement path.

     The experimental data show that those substituted phenols which are
capable of existing as a tautomeric equilibrium mixture are not detected
by the PH-2 instrument.  This may be due to the fact that the bathochromic
shift of these compounds lies outside the absorption maxima of phenol
and/or that, in a basic solution, the keto form is favored and thus gives
anomalous spectral results.  It is recommended that users of the instru-
ment be made aware of the anomalous results to be expected from substituted
phenols that can show tautomeric behavior.

     Although the Ministil (MS-1) was used in this work, no recommendation
is made for its use with the PH-2 instrument.  This unit was designed as
an adjunct to the PH-2 instrument for the unattended distillation of
precisely 50-ml samples.  This still is a very useful device but its
present cost prohibits any recommendation for its use instead of the
standard distillation equipment.  Further consideration should be given
to the application of this unit before a recommendation is made.

                               OBJECTIVES

     Most of the methods investigated for the analysis of phenolic com-
pounds in water have been found to have some deficiencies which limit
their application.  The spectrophotometric method shows the greatest
promise because of its reliability, ease and rapidity of measurement, and
possible adaptability to continuous monitoring.  Based on the knowledge
of the limitations encountered in other methods, a commercial spectro-
photometer has been developed and is now available for the determination
of phenolic compounds in water.  The instrumental technique takes advan-
tage of the bathochromic shift in phenolic compounds, which allows for
a quantitative measure of phenolic substance.

     Thus, the major objective of this project was to evaluate the
utility of the recently available commercial instrument designed specifi-
cally for the determination of phenolic compounds in water and to study
the instrumental and experimental parameters so that any improvements in
sensitivity and reliability of analysis of phenols can be realized.  An
added objective was to provide the resources, capability, and support of
the Department of Chemistry, Central State University, to the solution
of a problem confronting the management of water quality.

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                            INSTRUMENT DESIGN

     The instrument investigated is an ultraviolet ratio spectrometer
manufactured by Spectro Products, Inc., New Haven, Connecticut, and which
has the trade name, PH-2.  It consists of two conventional hollow cathode
lamps which, by means of a square-wave amplifier, are pulsed 180 degrees
out of phase, a monochromator, and a photomultiplier tube.  The hollow
cathode lamps are chosen so that light from one lamp is absorbed by the
solution under acidic conditions while the other lamp is insensitive under
these conditions.  When the solution is made basic, light from the one
lamp is absorbed and the other lamp is insensitive.  The difference in
the absorption under the acidic and basic conditions is proportional to
the concentration of phenolic compound present.  Because the instrument
does not differentiate between various types of phenolic compounds, the
results are reported as phenol.  Fortunately, the extent of the batho-
chromic shift of phenolic compounds is independent of the concentration
of the base; therefore, a pH of approximately 12 is sufficient to effect
the shift and permit a quantitative measure of the phenolate ion.

     During the evaluation, problems were encountered because the meter
needle drifted inordinately during a given reading.  This problem was
corrected when the manufacturer improved the electronic circuit of the
instrument, and no drift has been noted over an eight-hour period.  The
noise in the instrument can be decreased by use of the damping control
without any significant loss in accuracy.  The instrument is capable of
increasing the sensitivity of low readings by converting the full scale
to 0-10%, but more care and attention must be given to balancing the
lamps.  Thus, more time is required if one operates at the higher sensi-
tivity setting, but the time required for an analysis can still be less
than five minutes.

     This method requires as reagents only an acid and a base.   After any
necessary distillation, the sample is adjusted by a drop of concentrated
H3p04 to pH 4.  The solution is placed in the absorption cell and the
proper lamp is adjusted to read zero.  The cell is removed and the solu-
tion is adjusted to pH 12 by the addition of a base.   It has been found
that two drops of saturated sodium hydroxide are sufficient to bring the
solution to a pH of 12.  The cell and contents are returned to the spect-
rometer and the reading is observed.  As a precautionary measure,  it is
important that the cells be washed thoroughly after each reading to avoid
any cross contamination.  The concentration of phenol may be determined
directly from a standard curve which may be prepared, or by calculation
from the equation:

            Phenol, ppm = K log  100-meter reading
                                      100

where K is the calibration constant.  The equation is a form of the Beer-
Lambert law as related to the specifics of the instrument and of phenol.

-------
                         APPARATUS AND REAGENTS

     Reagent grade chemicals were used throughout this study, and they
were purchased from a single source.

     All solutions were prepared using phenol-free water.  The phenol-free
water was prepared by passing distilled water through a glass column
packed with a 4-cm X 25-cm column of 8-10 mesh activated animal charcoal.
The phenol-free water was checked periodically, and the column was re-
packed whenever the blank water showed a phenol content of over 5 ppb.

     The prepared stock solutions of phenol and substituted phenols
(0.5 g/500 ml) were stored in a refrigerator.  Fresh stock solutions were
prepared after two weeks, if the stock solution had not been depleted.
All other solutions were prepared fresh on the day of use by the appro-
priate dilution of the stock solutions.

     The solutions of ammonium chloride, 4-AAP, and potassium ferricyanide
for the color reaction in the 4-AAP method were prepared fresh each day
in accordance with the procedures described in American Society for Testing
and Materials (ASTM) and American Public Health Association  (APHA) manuals
(27, 28) for the direct photometric procedure and the chloroform extrac-
tion procedure.

     All pH measurements were made with a Sargent-Welch Model PEL pH meter
equipped with a combination electrode and standardized with a commercial
pH 10 buffer.

     All colorimetric measurements made by the 4-AAP method were made with
a Bausch and Lomb Model 70 spectrophotometer.  Initially, the response
of the instrument was checked by preparing a Beer's law plot of standard
sodium dichromate solution.  The PH-2 instrument was used in accordance
with the manufacturer's instructions.

     All glassware used in the study was maintained in accordance with
the procedure outlined in the quality control manual prepared by the
Environmental Protection Agency  (45).

     The standard water distillation apparatus with ground glass joints
was used.  In addition, distillations were carried out with Ministils
(MS-1) manufactured by Spectro Products, Inc.; these units provided for
the unattended distillation of 50-ml samples and were designed for use
with the PH-2 instrument.

     The various industrial  (real)  samples obtained from several coopera-
ting agencies were analyzed within  48 hours of the collection day and
were kept refrigerated.  All the industrial samples were preserved with
copper sulfate and phosphoric acid  in accordance with the recommended
procedures  (45) and were distilled  prior to analysis.

                                    10

-------
                              EXPERIMENTAL

     To evaluate this instrument objectively, the initial work involved
obtaining a thorough study of and familiarity with the 4-AAP method and
with the operation of the PH-2 instrument.  As recommended by the ASTM
and APHA, the 4-AAP method involves two procedures—the chloroform
extraction and direct photometry.  The first procedure is used for samples
containing less than 0.1 mg/1 (100 ppb) of phenol and the latter for those
containing more than 0.1 mg/1 (100 ppb).  The recommended upper limit for
analysis by the direct photometric procedure is 5 mg/1, but appropriate
dilution of samples may be made to remain within the recommended concen-
tration range.

     The initial work consisted of the application of both of the 4-AAP
procedures to the analysis of synthetic samples of phenol and the prepara-
tion of a working curve for both procedures.  In Table I, data are pre-
sented to show the results of the analysis of phenol solutions by the
chloroform extraction and the direct photometric procedures.  These results
represent a large number of samples that were analyzed over an extended
period of time by three different persons.  To approximate real conditions,
each analyst prepared his/her own solutions from a single stock solution
and worked independently.  In this manner, systematic errors in the tech-
niques of the three analysts would average out.  The wide range of the
calculated standard deviation may be attributed largely to the variation
in the rate of reaction of phenol with 4-AAP to form the colored product.
As indicated previously, several reaction parameters, especially pH, time
of color development, and temperature, appear to have significant effects
on the analysis (31).  It is significant that the smallest deviation
occurs with those samples whose absorbance is in the 0.3-0.7 range
(50-20%T).  The variation in the amount of phenol found by analysis and
in the standard deviation indicates the reliability of these two pro-
cedures.  In agreement with the work of other investigators, the chloro-
form extraction procedure appears to be the more sensitive.

     For routine determination of phenols in water,  a working curve can
be prepared and the concentration determined directly from it.  This
procedure is a time-saving device which can be used in laboratories where
a large number of analyses are determined.  However, the working curves
must be checked periodically because of changes with time of the spectro-
photometer.  Using the data given in Table II working curves were prepared
for the chloroform extraction and direct photometric procedures (Figures
1 and 2).

     The evaluation of the PH-2 instrument was undertaken after a pre-
liminary study was made of its operating parameters.  Unlike the 4-AAP
method, the PH-2 determination is not dependent on a color reaction but
on the bathochromic discussed earlier.  The method is thus independent of
any substituents on the ring which have been shown to interfere in the
analysis by the 4-AAP method.  The only reagents needed are concentrated


                                   11

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                          Table_I_

Phenolic Concentration Values Determined by the 4-AAP Method

                Chloroform Extraction Procedure
Number of
measurements
8
16
24
26
24
16
8
8
Calculated
cone. (yg/1)
5
10
20
30
40
50
60
70
Average cone.
observed (yg/1)
5.28
9.82
18.58
30.47
39.81
49.89
61.32
72.67
Standard
deviation
±0.94
±0.38
±1.10
±1.79
±0.52
±0.70
±1.03
±2.80
Direct Photometric Procedure
33
45
36
30
36
1.0 mg/1
2.0
3.0
4.0
5.0
0.95
1.92
2.84
4.00
5.05
±0.52
±0.85
±0.67
±0.45
±0.70
                          Table II
         4-AAP Method - Preparation of Working Curve
               Chloroform Extraction Procedure
 Number of
measurements

     4
     4
     4
     4
     4
     6
     6
     6
     6
     6
Calculated
cone.(yg/1)
    10
    20
    30
    40
    50
Average cone.
observed(yg/1)

      9.97
     19.06
     28.92
     39.97
     49.95
                  Direct Photometric Procedure
     1.0 mg/1
     2.0
     3.0
     4.0
     5.0
      1.
      2.
      3.
      3.
01
03
00
99
      5.10
          Standard
          deviation

            ±0.04
            ±1.06
            ±0.18
            ±0.22
            ±0.09
±0.02
±0.08
±0.15
±0.11
±0.15
                             12

-------
    1.0-
CD
<
     10-
    -'*-M
    .01
               I     I   \   I  I
n
 10
I   I   I  I  I  IT
                                                            100
          FIGURE 1. 4-AAP METHOD  (CHCI3 EXTRACTION)
                              13

-------
    10-
              J	I    I  I  I  I  I I I	I    I	I  I  I  I  I I
co  1.0-
OQ
    0.1-
               I    I    I
II        I     I   I
                                 10

                               mg/l


        FIGURE 2. 4-AAP METHOD (DIRECT PHOTOMETRIC)
                           100
                              14

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phosphoric acid and a saturated solution of sodium hydroxide.  Studies
have shown that the pH is not critical, as long as the acidity of the
phenol solution is lowered to about pH 4.0 and the basicity is raised to
about pH 12.0.  The absorption cell containing the test solution is placed
in the light path and the lamp adjusted to read zero.  The cell is removed
and sufficient base is added to bring the pH of the solution to about 12.0.
The meter reading is applied to the following equation
             ,    , ,   v    „ .   100-meter reading
            phenol (ppm) = -K log          - «•
                       K = calibration constant

to determine the concentration of phenol.  Alternatively, a working curve
can be prepared by plotting the meter reading against concentration, and
the concentration of a sample determined from the curve.   Two absorption
cells are available; one cell has a path length of 2 cm for use in the
50 ppb to 50 ppm phenol concentration, and the other cell has a path length
of 10 cm for use in the lower than 50 ppb concentration range.  The instru-
ment has provision for scale expansion for use with the 10-cm cell which
is used in the lower concentration range.  It was found that in all analy-
ses, one drop of phosphoric acid was sufficient to shift the pH to 4.0,
and two drops of saturated solution of sodium hydroxide were sufficient
to shift the pH to 12.0.

     The calibration constant had been determined for the instrument and
reported to be 21.5.  However, because of the necessary electronic modi-
fications that had been made on this instrument by the manufacturer, the
calibration constants were redetermined (Table III) .  The calculated value
of 21.1 is in agreement with the value reported by the manufacturer for
the 2-cm cell.  The calculated value of the calibration constant of 4.32
for the 10-cm cell is in agreement with the reported value of 4.30.  These
calculated values were used in the calculation of the phenol concentration
of the subsequent work reported herein.  The calibration curves are shown
in Figure 3.

     Among the concerns of the evaluation of this instrumental method are
the accuracy and precision in the measurement of phenols, the simplicity
and rapidity of the measurements, and the ability to detect a wide range
of phenolic compounds.  It is these factors which limit the usefulness
of the 4-AAP method, and which this instrument is designed to overcome.

     A series of phenolic compounds was tested with the 4-AAP method and
the PH-2 instrument (Table IV) .   Most of the compounds selected were para-
substituted phenols, which are generally insensitive to the 4-AAP method.
Among the phenols chosen are those with alkyl, halogen, amino, hydroxyl,
nitro, and carbonyl substituents .  The selection of these various types
of substituents should provide a reasonable evaluation of the ability of
the PH-2 instrument to respond to a wide variety of phenols.  They include
those which are more likely to be found as pollutants, since most have
at least slight solubility in water.
                                   15

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                                Table III

      Data for Determination of Calibration Constant,  PH-2  Instrument
Phenol, cone.
mg/1
2
3
4
5
10
20
0.10
0.02
0.03
0.05
0.06
K
calibration constant
2-cm cell
21.23
21.03
21.38
21.82
20.21
21.03
10-cm cell
4.78
4.36
4.28
4.10
4.20
Average
deviation
±0.08
±0.18
±0.21
±0.37
±0.64
±0.24
±0.21
±0.12
±0.11
±0.27
±0.09
     As with other analytical methods for the detection of phenols,  the
PH-2 instrument is not capable of determining the concentration of specific
phenols.  The data obtained report the concentration of the various sub-
stances as phenol.  However, it is expected that some variation may occur
in the concentration of the substituted phenols because of the differences
in the bathochromic shift experienced by the various phenols.   Since the
use of hollow cathode lamps gives monochromatic light of a very narrow
wavelength, the maxima of absorbance of the phenols may fall outside the
range of the lamps selected.  Nonetheless, the results of this study show
some significant features.  Table IV shows the comparative data of the
analysis of various substituted phenols with the 4-AAP method and with the
PH-2 instrument.  According to previous work, phenols substituted in the
para position are insensitive to the 4-AAP method except those with halogen,
carboxyl, hydroxyl, methoxy, or sulfonyl substituents since these groups
are capable of being replaced in the oxidative reaction of 4-AAP with
phenol.  The data for p-hydroxybenzoic acid, p-chlorophenol, p-methoxy-
phenol, and 2,6-dichlorophenol support this observation.  The results
obtained for these phenols are low, but this may be due to several factors,
such as time for development of color and incomplete displacement reaction
in addition to the factor of shift of maxima mentioned above.   However,
as expected, the phenols having other substituents in the para position,
such as p-cresol, 3,4-dimethylphenol, tyrosine, vanillin, p-aminophenol,
p-tertbutylphenol and p-hydroxybenzaldehyde do not show any appreciable
reaction with the 4-AAP method.

     The response of the PH-2 instrument to these substituted phenols
shows significant features.  It will be noted that 4-nitrophenol shows no
                                    16

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100
                                                       100
                      CONC - x102ppb
                             x10°ppm
                FIGURE 3. PH-2 INSTRUMENT
                           17

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Table IV


Comparison of Methods with Substituted Phenols
Compound
phenol
p-cresol
m-cresol
2 , 6- dimethylphenol
3,4-dimethylphenol
4-nitrophenol
1- tyros ine
vanillin
Cone. , calc
mg/1
2.0
2.0
2.0
2.0
2.0
2.0
2.0
2.0
p-hydroxy benzoic acid 2.0
p-aminophenol **
p-chlorophenol
2 , 6-dichlorophenol
p-methoxyphenol
p-tert-butylphenol
ra-hydroxyacetophenone
hydroquinone
2 ,4-dichlorophenol
resorcinol
p-hydroxybenzaldehyde
2-naphthol
thiophenol
4-methylthiophenol***
* Analysis run in
2.0
2.0
2.0
2.0
4.0
2.0
2.0
2.0
2.0
2.0
2.0
2.0
2.0
triplicate;
Cone.
4-AAP
mg/1
2.05 ± 0.04
0.0
1.77 ± 0.03
0.94 ± 0.06
0.14 + 0.06
0.05 ± 0.14
0.27
0.03 ± 0.10
1.02 ± 0.08
0.05 ± 0.15
1.61 ± 0.25
0.97 ± 0.09
1.06 ± 0.12
0.20 ± 0.03
0.60 ± 0.07
0.22 ± 0.16
0.97 ± 0.11
1.01 ± 0.1
0.0
0.43 ± 0.15
0.00
,obs*
PH-2
mg/1
2.06 ± 0.02
1.94 ± 0.05
2.16 ± 0.03
2.01 ± 0.03
2.01 ± 0.08
****
1.99 ± 0.02
****
2.02 ± 0.02
1.58 ± 0.08
1.19 ± 0.05
2.00 ± 0.04
****
3.03 ± 0.04
****
****
0.25 ± 0.13
2.38 ± 0.11
****
1.75 ± 0.09
0.60 ± 0.12
1.36 ± 0.08 0.61 ± 0.04
average deviation determined
  ** Practical grade; used without purification
 *** Commercial grade
**** Instrument gave zero or negative reading for bathochromic
     shift
                            18

-------
appreciable reaction with 4-AAP and no response with the PH-2  instrument.
However, 4-nitrophenol is highly colored and this may lead to  inter-
ferences.  In addition, because of the color, the absorption maxima of
the phenol and phenolate ion may be expected to lie outside the range of
the instrument.  It is significant that, with a few exceptions, those
phenols which are unreactive in the 4-AAP method show positive results
with the PH-2 instrument.  The cresols and tyrosene and chlorinated phenols,
which are the most important pollutants in this group, are detected by
the PH-2 instrument.

     It is significant that the compounds, vanillin, p-methoxyphenol,
hydroquinone and p-hydroxybenzaldehyde gave no response with the PH-2
instrument.  Similar problems were experienced with these substances
with the 4-AAP method.  However, a closer examination of these compounds
shows that all contain a hydroxy or carbonyl substituent and, in general,
represent phenol compounds which are capable of existing as tautomers.
Considerable study has been done on the tautomeric equilibrium that may
exist with the substituted phenols in acidic and basic media (46, 47).
Since these specific substituted phenols can exist as a tautomeric equi-
libria, it is not surprising that they show an anomaly in their absorption
spectra, and this factor suggests the reason they do not respond to the
PH-2 instrument and behave as they do with the 4-AAP method.   Thus, if
these compounds exist as a keto-enol tautomeric equilibrium mixture, the
maxima of the equilibrium mixture probably would not lie in the same
absorption range as phenol.  The absorption spectra of these compounds
were run using the same concentration as shown in the data, and an examin-
ation of the spectra shown in the Appendix suggests that the anomalous
behavior observed would be expected.  In addition to the above specific
compounds, resorcinol and naphthol show a large deviation from the cal-
culated value, even though significant amounts of them were detected.
However, it has been shown that these exist as a tautomeric equilibrium
mixture, although the equilibrium greatly favors the enol form.  It might
be speculated that the significant deviation from the calculated value
may be due also to the existence of such an equilibrium mixture in these
compounds.

     In addition to the comparison of the two methods on individual
phenols, synthetic mixtures were prepared, and the results  of the two
methods were compared.  The data for the synthetic mixtures are shown in
Table V.  The concentrations of the individual components in  the mixtures
were arbitrarily chosen, but the prepared mixture was selected for spe-
cific reasons and conditions.   The mixture of phenol and cresols and
mixture of cresols and chlorophenol were chosen because they frequently
occur as pollutants.  The other mixtures were prepared based  on the
results of the analysis of the individual compounds.  Mixtures were pre-
pared to contain compounds which were (1) insensitive to the  4-AAP method
but detectable by the PH-2 instrument, (2) to contain compounds which
were sensitive to the 4-AAP method but insensitive to the PH-2 instrument
method, and (3) to contain compounds in which some were insensitive to one
method but not to the other.  The latter group of mixtures  permitted the
evaluation of both methods in detecting compounds in the presence of
                                    19

-------
extraneous compounds and to determine the influence on the analyses of
these extraneous compounds.  The results show good agreement with the
calculated amount of phenols.  It is uncertain if the analyses by the
PH-2 instrument method are affected by the presence of phenols that can
show the tautomeric effect.  Additional work could be done to ascertain
the effect of these tautomeric phenols, but at present, they do not
represent serious pollutants and have not been reported as pollutants in
water, even though they are water soluble.  The average deviation was
determined rather than standard deviation, because the samples were run
in triplicate only and sufficient number of analyses were not made to
warrant a greater statistical analysis.

     The accuracy of the PH-2 instrument was determined by preparing known
samples of phenol and adding known amounts of phenol ("spikes").  This
procedure provides a means of determining the amount of recovery of phenol
when added to another sample.  The samples prepared included the range of
concentration of parts per million to parts per billion of phenol.  The
range of concentrations used represent the range of detection of phenol
by the PH-2 instrument, including the  expanded scale.  The experimental
work involved initially the preparation and analysis of phenols of known
and varying concentrations.  An aliquot was removed from the initial sample
and an aliquot of a known concentration of phenol was added and the sample
again analyzed.  An appropriate correction was made for the amount of
sample withdrawn and the amount added.  In each instance, the aliquot
removed was equivalent to the amount added so that the initial and final
volumes remained constant.  Correction was made for the amount of phenol
removed.  All of these experiments were performed on phenol solutions and
several samples were "spiked" with p-cresol instead of phenol.  Also in-
cluded in this study was the addition  of "spikes" to several industrial
water samples that were collected and  analyzed.  The data for these studies
are shown in Table VI.  For these experiments, no comparable studies were
conducted with the use of the 4-AAP method, since the purpose of the study
was to determine the accuracy and reliability of the PH-2 instrument.  The
samples were run in triplicate and only the average deviation was deter-
mined.  The data show a high percent recovery of phenol which in turn
indicates the accuracy of the instrument in determining phenol over a
wide range of concentrations.

     In Table VII are shown data on the analysis of a variety of industrial
wastewater samples.  All of the samples were collected and preserved with
copper sulfate and phosphoric acid by  the agencies from whom the samples
were obtained.  The samples were analyzed within a 48-hour period after
collection.  All of the samples were kept refrigerated until the analyses
were completed.  The samples were collected monthly over a six-month
period, and thus represent a reasonable spectra of environmental condi-
tions .  The sampling sites included a  rural and municipal water treatment
plant, a foundry, a chemical manufacturing plant, an agricultural stream,
and downstream river sites.  The variety of the sampling sites provided
a possible wide range of phenol concentrations in water.  The samples were
analyzed by the 4-AAP method and by the PH-2 instrument method.  In some
instances, the sample size was not sufficient to run triplicate samples


                                    20

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                            Table V

Comparison of Results Obtained for Phenols in Distilled Water

                  by the 4-AAP and PH-2 Methods
Cone. Total cone.
Mixture (nig/1) mg/1 (calc)
I.
II.

III.
IV.
V.

VI.
VII.
phenol
p-cresol
m-cresol
p-cresol
m-cresol
p-chlorophenol
phenol
p -met hoxypheno 1
p-cresol
p-cresol
3,4 dimethylphenol
p - 1 er t - buty Iph eno 1
p- chloropheno 1
resorcinol
m-cresol
p-cresol
4-tert-butylphenol
3,4 dimethylphenol
p-methoxyphenol
hydroquinone
5.0
2.0
1.0 8.0
5.0
5.0
1.0 11.0
4.0
2.0
2.0 8.0
3.0
3.0
3.0 9.0
4.0
2.0
2.0 8.0
4.0
2.0
2.0 8.0
3.0
3.0
Total cone.
mg/1 (obs)
4-AAP PH-2
5.8 8.0

4.9 9.3
5.7 5.0
6.7 9.30

5.66 8.21
3.6 8.03

        m-hydroxy
          acetophenone
                   3.0
 VIII.
tyrosine           2.0
p-hydroxyzoic acid 2.0
p-tert-butylphenoi 2.0
9.0
                                        6.0
                                                          no
2.97     response
             1.4
            5.18
                               21

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                                Table VI
           Recovery of Phenol by the PH-2 Instrument Method


Sample
1
2
3
4
5
6
7
8
9
Phenol in
sample
mg/1
2.14
0.98
2.03
4.17
3.07
0.33
0.03
0.02
0.04
Phenol
added
mg/1
.08
1.80
2.40
0.60
0.90
0.04
0.04
.02
0.10
Phenol
found
mg/1
2.87
3.20
4.77
4.73
4.00
0.36
.06
.03
0.12


% Recovery
97
111
108
99
100
97
86
75
86
for both analytical procedures.  The values reported are average values.
With few exceptions, the PH-2 instrument data showed consistently higher
results than the 4-AAP method.  This result is not unexpected since the
previous work has demonstrated clearly that the PH-2 instrument will
detect phenols which are insensitive to the 4-AAP method of analysis.
This fact would suggest that these water samples contained substituted
phenols which would escape detection by the 4-AAP method.  No reasonable
explanation can be given for those results in which the 4-AAP analyses
were higher than with the PH-2 instrument, unless it can be assumed that
these samples contained phenols or other substances which enhanced the
4-AAP color reaction and thus gave anomalous results.  It is not reason-
able to expect that these samples contained substituted phenols which
would give anomalous results with the PH-2 instrument as shown above
with the substituted phenols which can exhibit tautomerism.

                               DISCUSSION

     The initial work of this project included a thorough review of the
literature of the various methods available for the analysis of phenols
in  water and wastewater.  An  evaluation of these methods indicates that
the 4-AAP method is the most  reliable and  less time-consuming of the
various colorimetric methods.  On the other^hand, it is  obvious that
it  has  several  serious limitations, primarily its dependence on reacting
a reagent with  the  aromatic ring of phenol to produce a  colored product
which  is measured  and related to the concentration of phenol.  The ultra-
violet  spectral method appears to overcome many of the  limitations of
these  other methods.  It  is dependent on the shift in absorption maxima
 (bathochromic  shift) as the phenol  in acid solution  is  converted to  the
phenolate  ion  in a basic medium.  Thus,  it is independent of the sub-
stituents  on the aromatic ring of phenol and  does  not rely on  any
 reaction of the ring.
                                    22

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                 Table VII

Phenol Concentration of Industrial Samples
	(Average Values)	

                           Concentration
                                yg/i
Sample                   4-AAP        PH-2

 120
 121
 122
 123
 124
 125
 126
 127
 128
 129
 130
 131
 132
 133
 134
 135
 136
 137
 138
 139
 140
 141
 142
 143
 144
 145
 146
 147
 148
 149
36.9
11.9
51.6
45.2
45.2
39.3
4.8
83.3
15.5
44.1
10.7
93.0
128.0
40.0
24.0
39.0
142.0
65.0
192.0
40.0
48.0
35.7
0.0
7.7
17.9
836.0
160.0
0.0
403.0
1480.0
48.6
19.7
25.7
47.9
47.0
40.5
31.0
93.8
19.6
46.8
10.8
14.3
196.0
94.0
94.0
230.0
380.0
141.0
240.0
94.0
85.0
58.0
14.0
49.0
88.0
832.0
230.0
18.0
954.0
3460.0
                     23

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     The experimental work encompassed evaluating the PH-2 and comparing
it with the 4-AAP method.  The initial work included the preparation
and analysis by the 4-AAP method of synthetic samples of phenol varying
in concentration from 0.01 to 5 mg/1.  The initial experimental work pro-
vided a thorough familiarity with the 4-AAP method.  Similar kinds of
experiments were conducted with the PH-2 instrument to gain a familiarity
with its operation and response.  Significantly, the PH-2 gave more
reproducible and more precise results than the 4-AAP method.  This work
on the comparison of the two methods was extended to a large variety of
substituted phenols.  The substituted phenols were selected to represent
the most frequently occurring phenols, those which have been reported as
pollutants, and those that have substituents  in the ortho-, meta-, and
para-positions of the aromatic ring.  This work clearly demonstrated the
limitation of the 4-AAP method in determining phenolic compounds with
certain substituents in the para-position.  The same phenols which are
not detected by the 4-AAP method are detected by the PH-2 instrument.

     However, it is significant that some substituted phenols are not
detectable by the PH-2 instrument.  It was found that hydroxyl or carbonyl
substituted phenols give anomalous results.  Also, it was found that some
of these specific phenols were not detected by the 4-AAP method.  A close
examination of the structure of these phenols show that they are capable
of existing as a tautomeric mixture, which may explain their anomalous
behavior.  An examination of the literature showed that in solution, the
tautomeric equilibrium mixture of these phenols favors the keto form.
Fortunately, most of these particular phenols have not been reported as
serious water pollutants.  The ultraviolet spectra of these phenols were
studied, and it was found that their absorption maxima lie outside the
range of the instrument and outside the wavelength used in the 4-AAP
method.  However, no attempt was made to develop a technique for their
measurement under the present experimental conditions, but if it should
become necessary to measure these phenols, a comprehensive study must
be undertaken.

     Experimental work was carried out to determine the accuracy of the
PH-2 instrument by adding phenol of a given concentration to a known
and measured concentration of phenol.  The amount of recovery was then
determined, and it was found that the instrument provided accurate
analyses.  Again, varying concentrations of phenol were used which
covered the range of the instrument.  A few experiments were conducted
which included the addition of a given concentration of a substituted
phenol to known concentrations of phenol.  These experiments, along with
the previous work demonstrated the reproducibility and reliability of
the instrumental method of analysis of phenol.

     An error analysis was made of the data obtained from the PH-2 instru-
ment to ascertain the accuracy of the measurements.  These data were
obtained as the difference between the observed and true values divided
by the true value with the result presented in percent.  In Table VI are
shown data for measurements of spiked samples with the use of the 2 cm
                                    24

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and the 10 cm cells designed for measurement in the 50 ppb-50ppm and
5 ppb-50 ppb range, respectively.  Analysis of the data shows a 1.57%
accuracy for the 50 ppb-50 ppm range and 10-25% accuracy for the 5-50 ppb
range.  The average value of the accuracy for the 50 ppb-50 ppm range is
±4.7%, and the average value for the 5-50 ppb range is ±11.9%.  These
values indicate that the range of accuracy for the PH-2 instrument is
acceptable.

     Although it was necessary to conduct experiments with known concen-
trations of phenol and substituted phenols, it was also necessary to
test the instrument with real samples.  Through the cooperation of a
local agency, industrial samples were made available.  They were analyzed
simultaneously by the 4-AAP method and the PH-2 instrument method.  In
nearly all instances, the PH-2 instrument gave higher results for the
phenol content.  This may be explained in either of two ways:  (1) sub-
stances were present which interfered and lowered the values obtained by
the 4-AAP method; or (2) the PH-2 instrument was capable of determining
phenols in the samples that were not detected by the 4-AAP method.  A
few samples did show higher results with the 4-AAP method and this may
be due to interfering substances that enhanced the color developed by
the 4-AAP method, or interfered with PH-2 instrument readings.  A more
comprehensive study would be needed to identify clearly the reasons for
the results.  However, it can be established that the PH-2 instrument is
capable of adequately analyzing wastewater samples.

     This work was limited to the evaluation of the PH-2 instrument and
no attempt was made to assess the cost projections for the instrument.
However, it should be mentioned that the instrument employs hollow cathode
lamps which are identical to those used in atomic absorption units, and
experience has shown that they have a life of about two years.   Other
maintenance costs should be comparable to any other ultraviolet spectro-
photometer.  The only reagents required are concentrated acid and satur-
ated solution of alkali.  For each analysis, one drop of concentrated
phosphoric acid and two drops of saturated sodium hydroxide solution are
sufficient.  Thus,  the cost of reagents is much less  than in other methods.
                                   25

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                                 REFERENCES

 1.   Cleaning  Our Environment:   The Chemical Basis for Action, ACS Special
     Sales  Issue,  American Chemical Society-; Washington, D.C.  (1969).

 2.   Brady,  N.  C., editor, Agriculture and the Quality of Our Environment,
     AAAS Publication 85,  American Association for the Advancement of
     Science,  Washington,  D.C.  (1967).

 3.   Ballinger, D. G., Environment Science Technology 1^, 612  (1967).

 4.   Sperry, K.,  Science 158,  351 (1967).

 5.   Marx,  L.,  Science rTO, 445 (1970).

 6.   Lake Erie Report, U.  S. Department of Interior, Federal Water Pollu-
     tion Control Administration, Washington, D.C.., August (1968).

 7.   Rosen,  A.  A., Sheel,  R. T., and Ettinger, J. Water Pollution Control
     Federation 35_, 777 (1963) .

 8.   Baker,  R.  A., J.  Water Pollution Control Federation 35, 1099 (1961).

 9.   Middleton, F. M.  and Lichtenberg, J.  J., Ind. Eng. Chemistry 52,
     99A (1960).

10.   Burttschell, R. R., Rosen, A. A., Middleton, F. M., and Ettinger, M. B.,
     J.  Am.  Water Works Assoc.  5±, 205 (1954).

11.   Boetius,  J., Medd Denmarks Fishlog Havundersdg N.S.I., 1^ (1954).

12.   Gibbs,  H. D., J. Biol Chem. 71., 445 (1927); Chem. Rev. 3_, 291  (1926),
     J.  Phys.  Chem. 31., 1053 (1927).

13.   Thereault, E. J., Ind. Eng. Chem. 21., 343  (1929).

14.   Ettinger, M. B. and Ruchhoft, C. C., Anal. Chem. 20, 1191 (1948).

15.   Beshgetoor, A. W., Greene, L. M. and Stenger, V. A., Anal. Chem. 16,
     694 (1944).

16.   Mohler, E. F. and Jacob, L. J., Anal. Chem. 29_,  1369 (1957).

17.   Stoughton, R. W., J.  Biol. Chem. 115, 293  (1936).
                                    26

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18.  Lykken, L., Treseder, R. S. and Zahn, V., Anal. Chem.  18_,  103  (1946)

19.  American Society for Testing and Materials, Method D-2580-68,
     Philadelphia, PA (1968).

20.  Baker, R. A. and Malo, B. A., Environment Science Technology^,
     997 (1967).

21.  Eichelberger, J. W., Dressman, R. C. and Longbottom, J. E.,
     Environment Science Technology 4-, 576 (1970) .

22.  Simard, R. G., Hasegawa, I., Bandaruk, W. and Headington,  C. E.,
     Anal.  Chem. ^3, 1384 (1951).

23.  Emerson, E., J. Org. Chem.  3_, 153 (1938).

24.  Emerson, E., Beacham, H. H. and Beegle,  L. C., J. Org. Chem. jj,
     417 (1953).

25.  Emerson, E., Kelly, K., Beacham, H. H. and Beegle, L. C., J. Org.
     Chem.  9_, 226 (1954).

26.  Emerson, E. and Kelly, K.,  J. Org. Chem. L3, 532 (1958).

27.  American Society for Testing and Materials, Method D-1783-70,
     Philadelphia, PA (1970).

28.  Standard Methods for the Examination of Water and Wastewater, 13th
     edition, Method 22-22E, American Public Health Association, New
     York,  NY (1971).

29.  Gottlieb, S. and Marsh, P.  B., Anal. Chem. lj$, 16 (1946).

30.  Rosenblatt, D. H.,  Denck, M. M. and Epstein, J., Anal. Chem. 26,
     1655 (1954).

31.  Ettinger, M. B., Ruchhoft,  C. C. and Lishka, R. J., Anal. Chem. 25,
     1783 (1951).

32.  Martin, R. W., Anal. Chem.  21., 1419 (1949).

33.  Shaw,  J. A., Anal.  Chem. 23_, 1788 (1951).

34.  Hirano, Y. and Tamura, T.,  Anal. Chem. 36_, 800 (1964).

35.  Friestad, H. 0., Ott, D. E. and Gunther, F. A., Anal. Chem. 41,
     1750 (1969).

36.  Murray, M. J., Anal. Chem.  21, 941 (1969).
                                     27

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37.   Schmauch, L.  J.  and Grubb, H. M., Anal. Chem. 26_, 308 (1954).

38.   Goldschmid, 0.,  Anal. Chem. 26^ 1421 (1954).

39.   Maranville, L. F. and Goldschmid, 0., Anal. Chem. 26^ 1423 (1955).

40.   Smullin, C. F. and Wetteran, F. P., Anal. Chem. 27_, 1836  (1955).

41.   Englis, D. T. and Wollerman, L. A., Anal. Chem. ^9, 1151  (1957).

42.   Wadelin, C. W.,  Anal. Chem. 28^, 1530 (1956).

43.   Mexler, A. S., Anal. Chem. 35_, 1936  (1963).

44.   Martin, J. M., Org, C. R., Kincannon, C. B. and Bishop, J. L.,
     J. Water Pollution Control Federation 39, 21  (1967).

45.   Handbook for Analytical Quality Control in Water and Wastewater
     Laboratories, Analytical Quality Control Laboratory, National
     Environmental Research Center, Cincinnati, Ohio  (1972).

46.   Michaelis, L., Chem. Rev.  1£, 243  (1935).

47.   Wheland, G. W., Advanced Organic Chemistry, 3rd edition,  John Wiley
     and Sons, New York, chapter  14 (1960).
                                     28

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                                  APPENDIX 0
  44000      42000      40000
              35000             30000         25000
                 I    I  li   i   i    I   i   i  i   i
225   230  235   240
260    270  280
   MILLIMICRONS

 FIGURE 1. PHENOL
290  300     320   340    370  400
                                        29

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  44000
 .. i .  .
42000
40000
35000
30000
25000
  I      I     I    I
                            I      I     I
                                                                 pH 12.0
                                        I	I
                              I
225   230   235   240
             250    260    270   280   290  300
                         MILLIMICRONS


                   FIGURE  2.  HYDRQQUINONE
                                       320   340     370  400
                                          30

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44000       42000       40000
  il  ...... 1 1 1
              1 1 1 1 1 1 1  1 1 1 1     i     i
                                                35000              30000         25000
                                              i    I    i _ i   . i _ i — I — i — i — i — i — I
                                                                             I     I
225   230  235   240      250
                               260    270   280   290 300
                                 MILLIMICRONS

                               FIGURE 3. VANILLIN
320   340     370   400
                                         31

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 44000       42000      40000
                     35000
  30000
25000
       III         I        I      I      I     I
225   230   235   240
250     260    270   280   290  300
            MILLIMICRONS

    FIGURE 4. 4-METHOXYPHENOL
320   340    370  400
                                         32

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  44000
 42000
LI I I I I
40000
      35000
              30000
                25000
                i   I
       I
    I
          I
I
I    I
I
I
I
I
225   230   235  240
            250    260    270   280
                   MILLIMICRONS
                         290  300    320   340    370  400
                       FIGURE  5  . 4-HYDROXYBENZALDEHYDE
                                        33

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440000
42000
40000
 35000
	I
 30000
L  I	
25000
                            OH
     230   235   240
            250     260    270   280  290  300     320   340
                       MILLIMICRONS


            FIGURE 6. 3-HYDROXYACETOPHENONE
                                                   370   400
                                       34

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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1.
4.
7.
9.
12
15
16
17.
REPORT NO. 2.
EPA-600/4-76-048
TITLE AND SUBTITLE
EVALUATION OF INSTRUMENT FOR THE
DETERMINATION OF PHENOL IN WATER
AUTHOR(S)
L. Shelbert Smith
PERFORMING ORGANIZATION NAME AND ADDRESS
Department of Chemistry
Central State University
Wilberforce, Ohio 45384
. SPONSORING AGENCY NAME AND ADDRESS
Environmental Monitoring and Support
Office of Research and Development
U.S. Environmental Protection Agencj
Cincinnati, Ohio 45268
.SUPPLEMENTARY NOTES
3. RECIPIENT'S ACCESSION- NO.
5. REPORT DATE
September 1976 ("Issuing date")
6. PERFORMING ORGANIZATION CODE
8. PERFORMING ORGANIZATION REPORT NO.
10. PROGRAM ELEMENT NO.
1HA-323-JBA-027
11. CONTRACT/GRANT NO.
R-803172-01
13. TYPE OF REPORT AND PERIOD COVERED
: Laboratory
14. SPONSORING AGENCY CODE
r EPA/ORD


. ABSTRACT
The Spectra Product's PH-2 Phenol Instrument was evaluated to determine
its applicability to water and wastewaters. The results show that the
determinations of phenol in water and wastewaters can be obtained with
an accuracy of ±5% in the range of 0.05 to 50.0 ppm and with an accuracy
of ±12% in the range of 0.005 to 0.05 ppm. The PH-2 instrument gave
more precise results than the 4-AAP methods and can be used to detect
para substituted phenols. As with the 4-AAP method, the PH-2 instrument
method cannot be used to detect all substituted phenols quantitatively.
KEY WORDS AND DOCUMENT ANALYSIS
a. DESCRIPTORS
phenol
ultraviolet spectrometers
water analysis
is.
DISTRIBUTION STATEMENT
!
\ release to public
b.lDENTIFIERS/OPEN ENDED TERMS
bathochromic shift
PH-2 instrument
phenolic compounds
19. SECURITY CLASS (This Report)
unclassified
20. SECURITY CLASS (This page)
unclassified
c. COSATI Field/Group
13B
7C
21. NO. OF PAGES
44
22. PRICE
EPA Form 2220-1 (9-73)
•fr U.S. GOVERNMENT PRINTING OFFICEH 976-657-695/613k Region  No.  5-11
         \
35

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