D. J.
Subach
and M. E. Butwill Bell
Texos ARM University
College Station, 77843
II An Analytical Procedure for the
Determination of Pesticides in Food
Since food supply quality and safety have become major
topics of concern, pesticide multiresidue methodology has
received considerable attention. With this in mind we
have devised a n experiment t o incorporate analysis of
some edible products fortified with selected chlorine-con-
taining organics. T h e procedure, a s patterned along the
work of Schnorhus and Phillips (I), is suitable for ad-
vanced level chemical analysis courses and can be com-
pleted within three t o four laboratory periods. This proce-
dure is most applicable t o small liheral arts college laho-
ratories, where class size is not a major obstacle.
In a n attempt to make the student more aware of this
concern, a highly practical laboratory exercise is described
for the nresentation of selected analvtical techniaues and
instrumkntation. T h e basis for this procedure is related t o
the necessity of maintaining a correlation between the for-
mal learning of independent analytical techniques and the
direct application of combined analytical techniques and
knowledge as. for examnle. in industrial chemical analv-
sis. hat is, a n attempt' has been made t o bridge the in-
dustrial-academic interface by emphasizing applied chem-
istry in the chemistry curricula.
Since there is a strong trend in this direction (Z),a n ef-
fort has been made t o demonstrate what a pesticide resi-
due chemist might d o in industry and how his job is ac-
complished by utilizing a specific experiment. T h e pesti-
cide residue chemist must assess the method of analysis
- , since it is necessarv t o
a s t o reliahilitv and nracticalitv.
provide sufficient screening of possible contaminant prod-
ucts. The method must he c a ~ a b l eof analvzine - - laree-
numbers of samples with very high precision.
This procedure has the advantage of invoking the tech-
niques of extraction, isolation, and detection (identifica-
tion and quantitation). It also demonstrates the impor-
tance of seeking a universal method of analysis when a se-
ries of similar compounds are t o he analyzed. This is a
method which determines and detects all organic chemi-
cals of interest and any significant alteration products.
Experimental
Reagents
Reagent grade propylene carbonate, granular anhydrous sadi-
um sulfate, Florisil 60/100 mesh, reagent grade diethy1 ether, Pe-
troleum ether, acetone, heptane, Alz03 tic plates. and analytical
reagent grade silver nitrate.
Silver nitrate solution: Dissolve 0.5 g AgNOs in 5 ml of distilled
water, add 100 ml of 2-phenoxyethanol (practical), and make up
to 1 I with acetone. Add 3 drops of 30% hydrogen peroxide. m k
and transfer to a dark (brown) colored glass bottle.
Procedure
The analytical operations consist of six steps: A small represen-
tative sample is prepared; the sample is extracted with suitahle
solvent; filtration is employed to separate the solvent and insalu-
ble material; isolation is accomplished by ealumn chromatogra-
phy; the pesticide is concentrated by evaporation; and finally de-
teetion is done by gas chromatography and/or thin-layer chroma-
tography.
The procedure suitable for fruit, vegetables, grains, meats, and
dairy products, except milk, is as follows: A suitable amount of
product is prepared such that each student will obtain a 50-g por-
tion fortified with 100 wg of methoxychlor, dieldrin, heptachlor,
endrin, or any readily available organic chlorine pesticides. The
fortifying should consist of no more than two out of a possible
four, so the experiment may be based on technique rather than
tedium. The portions are labelled appropriately and distributed
to the students. The problem is to determine which one or what
combination of the compounds is present, by comparison to
known organic chlorine pesticide standard solutions.
Extraction
A 50-g fortified sample is macerated far 2 min with 80 ml of
propylene carbonate. (In place of the propylene carbonate, a 20%
methylene chloride in petroleum ether solution has been found
satisfactory (31.1 The macerate is vacuum-filtered through a
Biiehner funnel containing a %-in. layer of granular anhydrous
sodium sulfate. The filtered extract is dried over 20 g of anhy-
dmus sodium sulfate for 5 hr.
Cleanup
Florisil, 60/100 mesh, is activated hy heating at 130'C for 1d in
an oven. A chromatographic column is prepared by adding 20 g of
activated Florisil to a ealumn 1 X 16 in. containing about 10 g of
anhydrous sodium sulfate. Additional anhydrous sodium sulfate,
5-10 g, is placed an top of the Florisil. The column effluent equiv
alent to 3 g of sample is transferred to the column and time is al-
lowed for penetration of the Florisil. Sample elution is performed
using a 200-ml portion of 7% diethyl ether in petroleum ether. A
sample standard internally fortified with a known concentration
of pesticide should, in general, he carried through the procedure
to determine the actual recovery. Prior to detection the eluted
volume is concentrated down to 5 ml by evaporation.
Detection
Detection can be made using conventional gas chromatography
or thin-layer chromatography. If gas chromatography is utilized,
a Y' in. X 6 ft stainless steel column of 10% SE-30 on 80-100 mesh
Chromosorh W can be used. The temperature for the detector,
column, and inlet are, respectively, 280, 190, and 260°C. Flow is
adjusted to give maximum sensitivity.
For tlc, aluminum oxide plates can be prepared according to
Kovacs ( 4 ) or purchased. The AlzOs plates are activated far 4 hr
at 200°C prior to their use. Spotting is performed using a 1-@l
pipet. The sample is spotted in varying concentrations each alter-
nate spot being a known standard. This will aid quantitation by
visual camoarison. (The techniaue
nature.) ~ k n an
.~will he semi-auantitative in
~ ~~~~~~
g 8 ' x 8 in. plate, spotting should be done 1 in.
~.~~~~~~~~~
~~ ~ ~~~
from the bottom and development should proceed to 1 in. from
the top. A 2% acetone in heptane mixture is used as the devel-
oping solvent. Developing time will he about 12-11 min. The
plate is removed from the developing trough when the solvent
front reaches a pencil marked point 1 in. from the top edge of the
plate, and is allowed to dry for 5-10 min. ~h~ plate is uniformly
sprayed with silver nitrate reagent and dried in a forced draft
oven (80°C) for 2 min. The plate is then expased to ultraviolet
.. . lieht
lieht for 15 min in a closed chamber. Soots will aooear. . to
d&k brown.
Recovery of Pesticides Added to Parsley and Green Beans
Prior to Extraction with Propylene Carbonate
cbmmodity ort tine at ion ppm
compound Residue Found 7o
Apparent Corrected Raovery
parsley none
endrin ii o m
0.98 0.w
0.93 93
~ r r e n~ e a n s none O.W OW
hoptachlor i.0 0.85 0.85 85
Volume 50, Number 12, December 1973 / 855
Discussion
A minimum recovery of 80% should he achieved using
this procedure as verified by Schnorhus and Phillips (I).
E x a m ~ l e sof a c t u d results are eiven in the table. Schnor-
bus and Phillips demonstrate &at regardless of the pesti-
cide compound. recovery remains consistent which allows
selection-of the pesticide from a large number of com-
pounds:
The application of gas chromatography will require that
not more than 0.2% propylene carbonate he present in in-
jected solutions. Use of Florisil as described will separate
pesticides from a maximum of 7-ml aliquots of propylene
carbonate.
This procedure might also he utilized with some pro-
duce ohtained directly from the supermarket. For exam-
.
~ l .e several
. tomatoes might be ohtained and this Droce-
dure run to determine the amount of pesticide contained
in them. According to the FDA specifications (5) the tol-
856 /Journal of Chemical Education
erance of chlordane is 0.3 ppm, endrin is zero and hepta-
chlor is also zero in tomatoes. An analysis, using the out-
lined procedure, on tomatoes, or other produce, for a se-
lected pesticide should prove interesting and informative,
and demonstrates the practicality of this experiment.
Acknowledgment
It is a pleasure to acknowledge the support of the Ther-
modynamics Research Center staff in the preparation of
this manuscript. The valuable suggestions and criticisms
of Professors B. J. Zwolinski, R. C. Wilhoit, R. D. Whea-
ly, and Mr. C. H. Dickson are greatly appreciated.
Literature Cited
(a) Kovscr,M. F.. J A a s o c . Of/ie.Agr. Chemists. 1&.8M(19631.
(5) Official FDATo1eranees.N. A. C. News. 27, (31, Jan./Feb., 1969.