arby nawangsari: 22 Des 2009

I. A. K. AFRIDI*
Z. PARVEEN*
S. Z. MASUD*
SUMMARY: This paper presents results of the effects of studied pesticides on amino acids in cotton seeds
and wheat grains stored at the room temperature (30 ± 3°C) after pesticide treatment for one month. Each
sample was analyzed in triplicate along with a control sample. The influence on amino acids, was found to be
variable and significant. The analytical data, obtained by employing amino acid analyzer, showed significant
quantitative variations in amino acids of both the food commodities.
Key Words: Pesticide effects, amino acids.
Chemistry
INTRODUCTION
Many scientists have studied the effects of different
pesticides on total protein in cotton seeds, wheat
grains and in other crops such as maize and soybeans
and found quantitative variations (1-8). The nutritive
value of protein and proteinaceous foods and food
products depends on their amino acids composition
and essential amino acids balance. Effects of some
herbicides on amino acids in soybeans have been
studied and it was reported that lysine increased without
reducing the amount of total protein (9). In another
study, the action of different herbicides on quantitative
and qualitative changes in soils was compared (10).
Wolfson and Shearer (11) investigated variations in the
amino acid composition in grain protein of maize grown
with or without pesticides and standard commercial fertilizers.
Ciszewska et. al. (12) tested six wheat varieties
for effects of different herbicides, and found that results
of protein and lysine varied depending on the year and
the cultivars. Not much work has been reported about
the effects of pesticides on amino acids in cotton seeds
and wheat grains during storage.
Effects of certain organochlorine (OC), organophosphorus
(OP), and synthetic pyrethroid (SP) pesticides,
namely, p,p'-DDT, monocrotophos and cyhalothrin
respectively on cotton seeds and OP compounds
(chlorpyriphos-methyl and pirimiphos-methyl) and SP
one compound namely permethrin on wheat grains
have been studied, evaluated and their results are presented
in this paper. The results were assessed using
analytical data from trial application of studied pesticides
directly to the aforesaid food commodities, stored
for one month in sealed glass jars. Each sample was
analyzed for amino acids in triplicate along with control
sample by employing amino acid analyzer.
* From Department of Chemistry, University of Karachi, Karachi-75270,
Pakistan.
ORGANOCHLORINE, ORGANOPHOSPHORUS
AND SYNTHETIC PYRETHROID PESTICIDES
AFFECTING AMINO ACIDS IN COTTON SEEDS
AND WHEAT GRAIN DURING STORAGE
Journal of Islamic Academy of Sciences 8:1, 47-52, 1995 47
48 Journal of Islamic Academy of Sciences 8:1, 47-52, 1995
EFFECTS OF PESTICIDES ON AMINO ACIDS AFRIDI, PARVEEN, MASUD
MATERIALS AND METHODS
The analytical method of Osborne and Voogt (13) was
adapted for the determination of proteins in cotton seeds and
wheat grains. The sample of cotton seeds was taken in three
replicates for each pesticide treatment and transferred to
glass jars. Calculated amounts of monocrotophos (2 ppm),
p,p'-DDT (2 ppm) and cyhalothrin (2 ppm) separately added to
the glass jars and shaken for 3 hours. samples of wheat grain
were also treated with three grain protectants namely chlorpyriphos-
methyl, pirimiphos methyl and permethrin at recommended
dosages i.e., 10, 4 and 2 ppm respectively. The
fortified samples of both the food commodities were allowed to
stand in the sealed glass wars for one month at room temperature
(30 ± 3°C). Each fortified sample was analyzed along
with the control sample. Cotton seeds and wheat grain samples
containing 10-12 mg protein were accurately weighed
and transferred to glass culture tubes and 10 ml 6N HCI was
added. The tubes were evacuated, and sealed. The samples
were hydrolyzed at 110°C for 24 hours and filtered. Filtrates
were evaporated to near dryness by a rotary vacuum evaporator
to remove HCI and the residues in each case were dissolved
in deionized water.
The Chromatographic glass column (10 mm i. d. x 200 mm
long) was prepared with a slurry of ion exchange resin Dowex
50 W-XB (100-200 mesh) in deionized water. A mixture of
studied amino acids which are commonly fond in cotton seeds
and wheat grains, was prepared in deionized water and quantitatively
transferred to the prepared column and allowed to
settle in the column bed. The column was eluted with deionized
water and then with 3.5% ammonia in deionized water.
For complete elution, fractions of 2 ml each were collected
and checked for the presence of amino acid with ninhydrin
reagent till violet color ceased to appear. All 2 ml fractions
were then combined and evaporated nearly to dryness by a
rotary vacuum evaporator to remove ammonia and taken up in
deionized water for amino acid analyzer. A suitable aliquot of
each concentrated volume was subjected to the system of
amino acid analyzed and quantitative recovery of each studied
amino acid was determined.
The concentrated extracts of pesticide cotton seeds and
Table 1: %Age of studied amino acids in cotton seeds after one month treatment with pesticides.
S1
No
Name of amino acid Cotton seed
control
Cotton seed +
Cyhalothrin
Cotton seed +
p,p’ - DDT
Cotton seed +
monocrotophos
01 Alanine (Ala.) 1.20 ± 0.12 1.01 ± 0.12 0.42 ± 0.03 1.53 ± 0.27
02 Ammonia (Amm.) 0.66 ± 0.14 0.50 ± 0.03 0.12 ± 0.03 0.01 ± 0.00
03 Arginine (Arg.) 3.02 ± 0.29 1.88 ± 0.02 1.24 ± 0.10 2.48 ± 0.05
04 Aspartic acid (Asp.) 3.27 ± 0.02 2.17 ± 0.04 1.22 ± 0.02 2.24 ± 0.17
05 Cysteine (Cys.) 0.12 ± 0.04 0.07 ± 0.02 0.82 ± 0.02 0.58 ± 0.04
06 Glutamic acid (Glv.) 10.57 ± 0.10 0.11 ± 0.03 3.24 ± 0.02 12.49 ± 0.39
07 Glycine (Gyl.) 1.26 ± 0.14 1.24 ± 0.11 0.52 ± 0.03 1.02 ± 0.04
08 Histidine (Hist.) 1.26 ± 0.14 0.75 ± 0.02 0.75 ± 0.02 4.69 ± 0.23
09 Isoleucine (IIeu.) 0.71 ± 0.08 0.53 ± 0.01 NIL 0.09 ± 0.02
10 Leucine (Leu.) 1.61 ± 0.15 1.32 ± 0.06 1.03 ± 0.07 1.91 ± 0.14
11 Lycine (Lyc.) 1.25 ± 0.13 0.94 ± 0.01 7.77 ± 0.09 6.21 ± 0.21
12 Methionine (Meth.) 0.99 ± 0.10 0.17 ± 0.02 0.63 ± 0.02 0.06 ± 0.02
13 Phenylalanine (Phe.) 1.56 ± 0.09 1.15 ± 0.02 0.69 ± 0.02 1.01 ± 0.04
14 Proline (Pro.) 1.75 ± 0.04 1.75 ± 0.04 0.72 ± 0.02 NIL
15 Serine (Ser.) 1.36 ± 0.11 0.71 ± 0.03 0.40 ± 0.03 0.79 ± 0.10
16 Threonine (Thr.) 1.04 ± 0.03 0.74 ± 0.03 0.33 ± 0.02 0.92 ± 0.12
17 Tyrosine (Tyr.) 0.69 ± 0.09 0.49 ± 0.03 0.36 ± 0.03 0.87 ± 0.06
18 Valine (Val.) 1.17 ± 0.06 0.86 ± 0.07 0.03 ± 0.01 0.23 ± 0.02
TOTAL PROTEIN: 33.49 ± 0.11 16.89 ± 0.04 20.41 ± 0.03 37.13 ± 0.11
Journal of Islamic Academy of Sciences 8:1, 47-52, 1995
wheat grains in deionized water were quantitatively transferred
to the prepared column. Each extract was allowed to
absorb into column bed and then eluted with deionized water
to wash out unnecessary substances except amino acids and
then proceeded exactly as mentioned in the preceding paragraph
to get elute ready for amino acid analyzer. The amino
acid analyzer (Biotronik LC 6001 Germany) has an automatic
system with a sample injector and computing integrators for
peaks. The system involved photometric detection of ninhydrin
reacted amino acids. The photometer was attached to the
recorder, which in turn was attached to the computing integrator.
The hydrolyzed sample was dried, dissolved in the recommended
buffer solution and subjected to amino acid analysis.
Each of the 3 hydrolysates use analyzed using optimal parameters
for amino acid analyzer. A standard solution of amino
acids was used to calibrate the analyzer at least every 24
hours. The amount of amino acids in the sample was quantitatively
determined by comparison with the area of the samples
and the standard. Figures 1 and 2 show typical chromatograms
of cotton seeds and wheat grain treated with p,p'-
DDT and pirimiphos methyl respectively. Results of analysis
are presented, with standard error in Tables 1 and 2.
RESULTS AND DISCUSSION
No comprehensive study of the nutritional and germination
significance of different constituents in cotton
seeds and wheat grains have been carried out after
application of pesticides on these food commodities.
The present investigation focuses on the study of proteins
and particularly emphasizes the quantitative
measurement of amino acids susceptible to decomposition
during storage by the treatment of different types
of pesticides in the studied food commodities. The
49
EFFECTS OF PESTICIDES ON AMINO ACIDS AFRIDI, PARVEEN, MASUD
Figure 1: i) Chromatograms of a standard mixture containing 25n-moles each amino acid.
ii) Chromatograms of extracted amino acids from cotton seeds treated with pp'-DDT.
50 Journal of Islamic Academy of Sciences 8:1, 47-52, 1995
EFFECTS OF PESTICIDES ON AMINO ACIDS AFRIDI, PARVEEN, MASUD
amino acids estimated by amino acid analyzer are seventeen
in number including seven essential amino
acids. Tryptophan, destroyed by acid hydrolysis, could
not be determined. In most of the cases, it was noted
that the influence of pesticides on amino acids in both
the food commodities was remarkably significant. In
the treated samples under investigation in this laboratory,
the amino acids arginine, aspartic acid and glutamic
acid were estimated in much higher quantities
than other amino acids in cotton seeds, while in wheat
grain only glutamic acid and proline were present in
higher concentrations as shown in Tables 1 and 2.
In cotton seed samples treated with monocrotophos
(OP), proline was destroyed completely, while p,p'-DDT
(OC) left the same effect on isoleucine an essential
amino acid. Such effect was not observed in the samples
treated with cyhalothrin (SP), but in this case a
very significant influence was evident, showing marked
decrease in concentration of all the seventeen detected
amino acids as presented in Table 1 and Figure 1. Particularly,
the decrease was very significant in glutamic
acid and in an essential amino acid, methionine while
negligible and no effect was observed on proline and
glycine respectively. Thus, the overall decrease of
amino acids corresponded total protein by reducing it
to 16.6%. The aforesaid OC pesticide tends to increase
essential amino acids i.e. cysteine and lycine, in considerable
proportions while the rest of the amino acids
and the ammonia decrease in concentration; thus
13.08% reduction was estimated in total proteins. The
increasing tendency is very significant in seven amino
acids, particularly in cysteine, histidine and lycine, and
in appreciable amounts in alanine, glutamic acid,
leucine and tyrosine in the samples treated with
monocrotophos. Due to this reason, an increase of
3.64% in total proteins, was estimated in this case.
Generally, a reducing effect was observed on arginine,
aspartic acid, glycine, methionine, phenylalanine,
Table 2: % Age of studied amino acids in wheat grains after one month treatment with pesticides.
S1
No
Name of amino acid Wheat
control
Wheat + Chlorpyriphosmethyl
Wheat +
Pirimiphosmethyl
Wheat +
Permethrin
01 Alanine (Ala.) 0.51 ± 0.02 0.59 ± 0.04 0.69 ± 0.02 0.51 ± 0.02
02 Ammonia (Amm.) 0.08 ± 0.02 0.08 ± 0.01 0.08 ± 0.01 0.82 ± 0.04
03 Arginine (Arg.) 0.06 ± 0.01 1.37 ± 0.04 1.04 ± 0.04 0.66 ± 0.02
04 Aspartic acid (Asp.) 0.03 ± 0.02 0.01 ± 0.01 NIL 0.10 ± 0.02
05 Cysteine (Cys.) 0.10 ± 0.02 0.21 ± 0.02 0.17 ± 0.01 0.05 ± 0.01
06 Glutamic acid (Glv.) 2.73 ± 0.10 3.55 ± 0.10 3.68 ± 0.04 2.12 ± 0.05
07 Glycine (Gyl.) 0.79 ± 0.03 0.86 ± 0.03 0.81 ± 0.03 0.76 ± 0.03
08 Histidine (Hist.) 0.45 ± 0.02 0.82 ± 0.02 0.70 ± 0.02 0.61 ± 0.02
09 Isoleucine (IIeu.) 0.38 ± 0.02 0.45 ± 0.01 0.57 ± 0.02 0.38 ± 0.02
10 Leucine (Leu.) 1.16 ± 0.04 1.59 ± 0.04 1.42 ± 0.06 1.00 ± 0.03
11 Lycine (Lyc.) 0.44 ± 0.01 1.52 ± 0.06 1.71 ± 0.02 0.82 ± 0.03
12 Methionine (Meth.) 1.79 ± 0.05 0.62 ± 0.02 0.25 ± 0.01 0.54 ± 0.02
13 Phenylalanine (Phe.) 0.59 ± 0.02 1.01 ± 0.05 0.92 ± 0.03 0.82 ± 0.03
14 Proline (Pro.) 2.72 ± 0.06 2.61 ± 0.07 2.48 ± 0.04 1.62 ± 0.05
15 Serine (Ser.) 0.10 ± 0.01 0.25 ± 0.02 0.28 ± 0.02 0.25 ± 0.03
16 Threonine (Thr.) 0.10 ± 0.01 0.21 ± 0.02 0.17 ± 0.01 TRACES
17 Tyrosine (Tyr.) 0.22 ± 0.03 0.33 ± 0.02 0.36 ± 0.03 0.67 ± 0.02
18 Valine (Val.) 0.58 ± 0.03 0.79 ± 0.04 0.86 ± 0.03 0.39 ± 0.02
TOTAL PROTEIN: 12.83 ± 0.03 15.50 ± 0.03 16.19 ± 0.02 12.12 ± 0.03
Journal of Islamic Academy of Sciences 8:1, 47-52, 1995
serine, threonine, valine and also in ammonia in the
samples treated individually with aforesaid pesticide.
In the samples of wheat grain treated with two OPs
chlorpyriphos-methyl and pirimiphos-methyl pesticides
individually, the six essential and eight amino acids out
of a total seventeen tend to rise with pesticide concentration
as incorporated in analytical data in Table 2 and
Figure 2. Evidently, a considerable reduction and complete
destruction of aspartic acid occurred by the
effects of above mentioned OP pesticides. Deficiency
however, was observed only in methionine and proline.
Thus the total increase in amino acids was found to be
directly related to the increase of total proteins contained
in control sample by 2.67 and 3.36% respectively.
In case of permethrin (SP), the increase was
noted to be associated with arginine, aspartic acid, histidine,
serine, tyrosine, and essential amino acids lycine
and phenylalanine while no influence was observed on
alanine and isoleucine. The trace amount of threonine
and considerable decrease in concentration of leucine,
methionine, proline and valine confirms adverse effects
of permethrin on these amino acids. Insignificant influ-
51
EFFECTS OF PESTICIDES ON AMINO ACIDS AFRIDI, PARVEEN, MASUD
Figure 2: i) Chromatograms of a standard mixture containing 25n-moles each amino acid.
ii) Chromatograms of extracted amino acids from wheat grains treated with pirimiphos-methyl.
52 Journal of Islamic Academy of Sciences 8:1, 47-52, 1995
EFFECTS OF PESTICIDES ON AMINO ACIDS AFRIDI, PARVEEN, MASUD
ence was observed on the rest of the amino acids. Due
to balanced effects on the amino acids, overall reduction
in total protein occurred only by 0.71%.
It may be inferred from the analytical data that the
amino acids cysteine, glutamic acid, glycine, leucine,
methionine and threonine have undergone similar
adverse effects with SP pesticides on both the commodities.
All the three studied OP pesticides left beneficial
effects on alanine, cysteine, glutamic acid,
histidine, leucine, lycine and tyrosine, while adverse
effects were observed on aspartic acid, methionine and
proline in cotton seed and wheat grain. Methionine and
proline were the only two amino acids which decreased
quantitatively by the effects of all the six pesticides
studied in samples of both the food commodities.
The analytical data of protein is very significant for
both the food commodities as the nutritive value of protein
and proteinaceous foods and food products
depend on their amino acid composition and essential
amino acid balance. SP pesticide, cyhalothrin proved
to be a very effective chemical agent as it reduced
quantitatively all the detected amino acids except proline
in the samples of cotton seeds. On the other hand,
in wheat grain, effect of another SP pesticide namely,
permethrin on amino acids was variable as it caused
both decreasing and increasing affects on eight and
seven amino acids respectively.
The influence of OP pesticides i.e. chlorpyriphosmethyl
and pirimiphos-methyl on all the amino acids in
wheat grain was found to be quite identical, while
another OP pesticide, monocrotophos, produced
adverse effects on majority of the amino acids in cotton
seed.
The OC pesticide p,p'-DDT caused decreasing
effect on all the studied amino acids present in cotton
seed samples except cysteine and lycine, both of which
increased quantitatively.
It may, therefore, be concluded that both studied SP
pesticides leave more adverse effects than studied OC
compound and cause deficiency of amino acids in both
the studied food commodities while studied OP pesticides
are found to be much milder in effects. Thus the
significant effects described above on individual amino
acids show that the studied pesticides may affect the
commodities if the period of storage is extended.
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Correspondence:
Irshad A. K. Afridi
Department of Chemistry,
University of Karachi,
Karachi-75270,
PAKISTAN.

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