IDENTIFICATION OF PCB'S IN THE PRESENCE OF

DDT-TYPE COMPOUNDS USING LOW

TEMPERATURE LUMINESCENCE
J. T. BROWNRIGG AND A. W. HORNIG
Baird-Atomic, Inc.
125 Middlesex Turnpike
Bedford, Mass. 01730

Low temperature (77~ luminescence spectra of polychlorinated biphenyls

(PCB's), DDT-type compounds and mixtures suggest a simplified method for identifi-
cation and quantitation of aromatic pesticides with minimal prior sample treatment.
Spectral differences allow identification of PCB's in the presence of DDT and the
converse. Approximate detection limits, possible interferences and suggestions for
further work are discussed.

Polychlorinated biphenyls (PCB's) have emerged as ubiquitous environmental

pollutants having been widely used as plasticizers, lubricants, flame retardants, and
as heat transfer media (Peakall and Lincer 1970, Gustafson 1970). The presence of
P C B ' s in the environment is a matter of concern for two reasons. First, they have
been found to be highly toxic to certain marine animals. For example, exposure for
48 hours to 0.1 ppm of Aroclor | 1254 (a commercial PCB mixture) in sea water
produces 100% mortality in juvenile pink shrimp, and strongly depresses the rate of
shell growth (Duke et al. 1970) Second, P C B ' s have been found to interfere with
gas chromatographic determination of dichlorodiphenyltrichloroethane (DDT) and
related compounds, requiring a separation step which usually involves thin-layer or
column chromatography (Reynolds 1971).

The objective of the present investigation was to assess the applicability of low
temperature (77~ luminescence spectroscopy to the problem of identifying PCB's
in the presence of DDT-type compounds. Basic studies completed to date have
focused upon the documentation of important pesticide and PCB spectra, including
various mixtures of these. The results obtained are very encouraging and suggest
that a rapid and sensitive method for PCB determination can be developed using
this approach. Studies are in progress on the application of the proposed method to
sampling of natural waters. Further extension to residue analysis seems quite
possible.

Presented at the Third International Congress of Pesticide Chemistry (IUPAC), Helsinki, Finland,
1974.

Archives of Environmental Contamination 175

and Toxicology Vol. 4, 175-182 (1976)
9 1976 by Springer-Verlag New York Inc.
176 J. T. Brownrigg and A. W. Hornig

Experimental

T h e p , p ' - and o,p' -derivatives of DDD, DDE, and DDT were obtained from the
EPA Perrine Primate Laboratory, Perrine, Florida. Samples were designated "Re-
ference Standard" and had stated purities of 9 9 + % . These compounds were used
as received.

Commercial PCB mixtures studied were Monsanto Aroclors 1016, 1221, 1242,
1248, 1254, and 1260. Samples were provided by Dr. E. S. Tucker and Mr. W. B.
Papageorge of the Monsanto Company. Aroclor 1016 is similar to Aroclor 1242
except that isomers having five or more chlorine atoms have been removed (Nisbet
and Sarofim 1972).

Methylcyclohexane was Matheson, Coleman, and Bell "Spectroquality" grade.

Residual amounts of toluene were removed by column chromatography through
silica gel. Dichloromethane was Fisher "Spectranalyzed" material. This was usu-
ally of sufficient purity to be used as received, although distillation was sometimes
required to remove luminescent impurities.

Spectra were obtained using a Baird-Atomic Model SF-100 " F l u o r i s p e c "

fluorescence spectrofluorimeter and are not corrected for instrument response
characteristics.

Spectra of PCB's and DDT-type compounds. The spectra of PCB isomers,

Aroclors, and DDT derivatives are discussed in greater detail elsewhere (Brownrigg
et al. 1972, 1974). The excitation/emission spectra of a highly chlorinatedPCB
mixture, Aroclor 1254, are shown in Figure 1. For those Aroclors studied, absorp-
tion commences in the 290-300nm region, with phosphorescence maxima in the
440-480 nm region. Although the emission spectra of the more highly chlorinated
compounds are diffuse, the corresponding excitation spectra are generally struc-
tured. (The reverse is true of compounds with low chlorine content.) Combined
differences in the excitation/emission spectra are sufficient to enable those Aroclors
studied to be distinguished.

Spectra of the DDD and DDT compounds are very similar, and that o f p , p ' -DDT
is shown in Figure 2. The phosphorescence excitation spectra show characteristi-
cally narrow bands near 275 nm, and thus resemble the absorption spectrum of the
related hydrocarbon, diphenylmethane, as given by Berlman (1971). Phosphores-
cence intensities are approximately ten times weaker than those of Aroclors.

Emission f r o m p , p ' -DDE was at least 100 times weaker than the DDD or DDT
phosphorescence, and the observed emission may, in fact, be due to an impurity.
Our results do not agree with those reported by Moye and Winefordner (1965).

Detection Sensitivities. Using the highest available spectral resolution (two nm),
the noise-limited detection sensitivity for Aroclor 1254 in purified methyl-
 Identification of PCB's 177

I I I

oc

I I I
200 300 400 500 600
Wavelength (nanometers)

Fig. 1. Excitation/emission of Aroclor 1254 in methylcyclohexane (one ppm) at 77~

I I I

I I I
200 300 400 500 600
Wavelength (nanometers)
Fig. 2. Excitation/emission of p,p' -DDT in methylcyclohexane (ten ppm) at 77~
 178 J. T, Brownrigg and A. W. Hornig

cyclohexane is about two ppb, sensitivities on the order of one to ten ppb are
expected for other Aroclors. The corresponding value for DDT (and DDD) is about
30 ppb. Detection sensitivities could be improved by utilizing larger analyzing
bandwidths; this, however, would result in poorer resolution and therefore poor
specificity.

Working curves for Aroclors 1016, 1248, and 1254 in methylcyclohexane showed
that the phosphorescence intensity varies linearly with concentration below about
ten ppm. The corresponding curve forp,p' -DDT was linear below about 30 ppm.

Mixtures of Aroclor 1254 and p,p' -DDT. Mixtures were studied in which the
relative concentrations ranged from 0.1 to ten ppm in methylcyclohexane. Spectra
of a mixture having 0.5 ppm of each component are shown in Figure 3.

Each component can be measured by proper choice of wavelength and spectral

bandwidth. DDT is best measured free of Aroclor interference from the excitation
spectrum monitored near 380 rim. This is an emission wavelength where the DDT
phosphorescence is appreciable, but the Aroclor 1254 emission is very low. Excita-
tion of the mixture near 290 nm with bandwidths of less than about 20 nm largely
avoids excitation of DDT, giving almost pure Aroclor emission.

i t I

DDT

clor

i i, i
200 300 400 500 600
Wavelength (nanometers)

Fig. 3. Mixture of Aroclor 1254 and p,p' -DDT in methylcyclohexane (each 0.5 ppm) at
77~ Excitation spectrum monitoring emission at 380 nm; emission spectrum excited at 290
rim.
 Identification of PCB's 179

Plots of the Aroclor 1254 emission intensity show good linearity over the con-
centration range 0.1 to ten ppm, and DDT concentrations of comparable magnitude
do not appear to significantly affect linearity. Similar plots of the DDT intensity
were linear except for the solution containing one ppm of DDT and ten ppm of
Aroclor 1254, for which the intensity was about 50% less than that expected. This
was not true of the same mixture diluted ten-fold with methylcyclohexane, so that it
appears that DDT is quenched by relatively high concentrations of Aroclor. The
quenching mechanism involved is probably energy transfer: after absorbing light a
DDT molecule transfers this energy to a nearby PCB molecule, which has lower-
lying energy states. The PCB acceptor molecule subsequently emits the energy
accepted from DDT.

Aroclor 1254 can be detected in the presence of DDT concentrations approxi-

mately 100 times higher. DDT, having inherently weaker emission than Aroclor,
can probably be detected in the presence of about 20 times greater concentrations of
Aroclor 1254. However, for absolute concentrations of Aroclor 1254 above ten
ppm, there appears to be significant energy transfer quenching of DDT. Sample
dilution may overcome this problem if loss of sensitivity is not too great.

Water Studies. In order to estimate recoveries and detection sensitivities for

PCB/DDT compounds in water, one-L water samples were doped with ethanol
solutions of Aroclor 1254 and p,p' -DDT. Doped vCater samples were stirred for a
day with a Teflon-covered magnetic stirring bar. The organic material was ex-
tracted with two successive 75-ml volumes of dichloromethane. After drying with
anhydrous sodium sulfate, extracts were concentrated to volumes of less than 0.5
ml using a rotary evaporator. Methylcyclohexane was then added (usually five or
ten ml) and the sample luminescence measured at 77~ Sample concentrations
were determined by comparison of intensities with those of standards in methylcyc-
lohexane.

P u r e water. Recovery of Aroclor 1254 from one-L samples of pure (distilled)

water was about 70% at the one ppb level and about 80% at the ten ppb level.
Similar experiments with p,p' -DDT gave recoveries of about 50% at the ten ppb
level and about 70% at the 100 ppb level. For both materials, the first two extracts
combined contained 95% or more of the total amount extracted.

The difference in recovery between Aroclor 1254 and DDT is probably real but is
not presently understood. The observed losses are probably due mainly to adsorp-
tion on glass surfaces.

Natural water. Analogous experiments were conducted using 1.7-L samples of

raw river water. This water, although relatively unpolluted, was yellow-brown due
to the presence of natural organic matter and also contained suspended particulates.
No attempt was made to filter these particulates prior to doping. Recoveries for
both Aroclor 1254 a n d p , p ' -DDT from these waters were two or three times lower
than found for pure water. This is thought to be due to increased adsorption by
 180 J. T. Brownrigg and A. W. Hornig

particulates, but increased solubilization by humic materials may also be responsi-

ble (Wershaw et al. 1969). Figure 4 shows the spectra of Aroclor 1254 extracted
from fiver water (doped at 60 ppb) with 75 ml of dischloromethane; the evaporated
extract was dissolved in ten ml methylcyclohexane for low temperature analysis.

Detection sensitivities. Detection limits for Aroclor 1254 and p , p ' -DDT in
one-L volumes of pure water are estimated at 0.03 and 0.5 ppb respectively. (As
for standard solutions in methylcyclohexane, detection limits quoted here assume
use of highest spectral resolution.)

Detection limits for these same compounds in 1.7-L volumes of river water of the
type used here are approximately two and ten ppb, respectively. These higher
values are due to lower recoveries and the presence of a strong background
luminescence from natural substances. If these compounds, presumably polar
humic material (Shapiro 1957), were removed prior to analysis, detection limits
could probably be improved by a factor of ten. In this regard, a simple chromatog-
raphic clean-up step using silica gel might be of value (Blumer 1970, Johnson
1971). Detection limits could also be improved by analyzing larger volumes of
water, although solvent extraction ultimately becomes impractical. The use of flow
systems through adsorbent polymer resins, such as those used by Harvey (1973)
and Uthe et al. (1972), should be evaluated for ~ossible compatibility with
luminescence analysis.

t I I
200 300 400 500 600
Wavelength (nanorneters)

Fig. 4. Excitation/emission of river water extract in methylcyclohexane at 77~ Water

sample had been doped with 60 ppb of Aroclor 1254.
 Identification of PCB's 181

Apart from natural compounds present in water, phthalic acid esters (phthalates)
may also be present. These compounds have been widely used as plasticizers,
particularly in the manufacture of polyvinyl chloride (Graham 1973). Two phtha-
lates have been studied to date, namely, dibutyl phthalate and diisodecyl phthalate.
The phosphorescence emission of these compounds resembles that of the more
highly chlorinated Aroclors. The excitation spectra are also similar, except for the
lack of secondary adsorption in the 250-260 nm region. Spectra of additional
phthalates need to be documented; those studied to date, however, suggest that they
might interfere with the determination of certain PCB's.

Conclusions
PCB and DDT concentrations at sub-part per billion levels in water can be rapidly
estimated using an extraction procedure followed by luminescence measurement at
77~ Detection limits in natural water samples having high levels of fluorescent
materials and particulates may be more than an order of magnitude higher. These
limitations could probably be overcome by the inclusion of a simple chromatog-
raphic step or the use of improved extraction techniques. The method described
utilizes commercially available instrumentation and could be useful as a screening
method in either a mobile laboratory or in a small field station. Although the
procedure described here does not include coupling with gas or liquid chromatog-
raphy, low temperature luminescence analysis might be used profitably with either
procedure for confirmatory measurements. The method may be extended to include
a wider array of aromatics.

Acknowledgments

The authors thank Mr. Dwight G. Ballinger of the Methods Development Quality
Assurance Research Laboratory, National Environmental Research Center, U.S.
Environmental Protection Agency, Cincinnati, Ohio, for his interest and encour-
agement.

We wish to also thank Dr. E. S. Tucker and Mr. W. B. Papageorge of Monsanto

for providing Aroclor samples. Assistance with the laboratory studies was provided
by Dr. Bonnie Dalzell and Mrs. Geraldine Garnick.

This work was supported in part by the U.S. Environmental Protection Agency
under Contract No. 68-01-0082.

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 182 J. T. Brownrigg and A. W. Hornig

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Manuscript received September 30, 1974; accepted November 23, 1974.