Eur. J. Lipid Sci. Technol.

2009, 111, 593–598 593

Research Paper
Acephate, methamidophos and monocrotophos residues in a
laboratory-scale oil refining process

Chee Beng Yeoh and Chiew Let Chong

Malaysian Palm Oil Board, No. 6, Persiaran Institusi, Selangor, Malaysia

Acephate, methamidophos and monocrotophos are insecticides used in oil palm plantations for the control
of bagworms and leaf-eating caterpillars. The main purpose of this study was to determine whether the
physical refining process at laboratory scale, which simulated the manufacturing process, could remove
the residues of these three insecticides in crude palm oil, in the unlikely event that crude palm oil were
contaminated with these organophosphorus insecticides. A series of crude palm oil samples spiked with
low (0.1 mg/g) and high (1.0 mg/g) levels of these insecticides were subjected to a laboratory-scale physical
oil refining process. Oil samples drawn at various stages of the refining process, namely, degumming,
bleaching and deodorization, were analyzed using an in-house analytical method. The results obtained
from these experiments suggest that the physical refining process is capable of effectively removing resi-
dual insecticides from crude palm oil. The final product of crude palm oil refining, the refined, bleached
and deodorized palm oil, was found to have no detectable levels of acephate, methamidophos and mono-
crotophos.

Keywords: Acephate / Methamidophos / Monocrotophos / Palm oil / Refining

Received: November 9, 2008; accepted: January 29, 2009
DOI 10.1002/ejlt.200800262

1 Introduction Vegetable oil refining can be carried out physically or

chemically. In the Malaysian palm oil industry, all crude
Oil palm is a major commercial crop in Malaysia. In the year palm oil is refined physically. The process begins with the
2007, the total oil palm plantation area in Malaysia was 4.31 degumming step where phosphoric acid is added to the
million hectares [1]. Similar to other crops, oil palms are sub- crude palm oil at ,90 7C. Next, the degummed oil is
jected to pest attacks. In order to counter this problem and pumped into the bleacher where the bleaching process
maintain oil palm productivity, integrated pest management occurs. Bleaching involves the addition of bleaching earth
(IPM) is adopted in most of the Malaysian plantations. How- (1–2% of oil weight) to remove undesirable impurities (pig-
ever, in the event of severe pest outbreaks where the IPM ments, trace metals, oxidation products) from crude palm
strategy fails to control the problem, pesticides are used as oil at a temperature ranging from 100 to 110 7C for 30 min.
preferred alternatives to protect the plants. The use of pesti- This degummed bleached oil is then filtered to remove the
cides in oil palm plantations has evoked concerns among gums and bleaching earth before it is pumped to the deo-
consumers, the authorities as well as palm oil producers dorizer. Deodorization is basically a vacuum-steam distilla-
regarding their effects on human health. tion process at an elevated temperature (250–260 7C) dur-
Crude palm oil is produced from the mesocarp of palm ing which free fatty acids and volatile compounds are
fruits. The orange-red crude palm oil is refined to produce a removed to produce an odorless and bland oil. In addition
clear, bland-flavored, odorless and light yellow product which to this, the carotenoid compounds are destroyed during the
meets the consumers’ expectations of an edible oil. heat treatment process, and this results in the final light
yellow, fully refined oil.
Over the past few decades, there were relatively few
Correspondence: Chee Beng Yeoh, Malaysian Palm Oil Board, No. 6,
reports regarding studies on the fate of pesticide residues
Persiaran Institusi, Bangi 43000, Selangor, Malaysia. during vegetable oil processing. Smith et al. [2] reported
E-mail: [email protected] that chemical refining was effective in removing chlorinated
Fax: 160 3 89259658 pesticides (endrin, DDT, DDE, aldrin, dieldrin, heptachlor

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594 C. B. Yeoh and C. L. Chong Eur. J. Lipid Sci. Technol. 2009, 111, 593–598

and heptachlor epoxide) in simulated commercial refining This paper investigates the fate of three organophos-
process experiments with crude soybean oil and cottonseed phorus insecticides in palm oil, namely, acephate, methami-
oil. They postulated that the removal of these pesticides was dophos and monocrotophos, at the different stages of a labo-
achieved by volatilization during the deodorizing step. The ratory-scale physical refining process which simulates the
same observation was reported by Mounts et al. [3] a few commercial process.
months later through their radiochemical experiments with
soybean oil (14C-endrin and 14C-benzo(a)pyrene). The
observation on the failure of the bleaching step in the 2 Materials and methods
removal of endrin confirmed the hypothesis suggested by
Smith et al. [2]. 2.1 Reagents and materials
On the other hand, Mounts et al. [3] reported the removal of
benzo(a)pyrene during the bleaching step (with activated car- All solvents used were of HPLC grade and were used as
bon), suggesting that a slight modification of the commercial received. Acephate, methamidophos and monocrotophos
vegetable oil processing procedures could be effective in remov- reference standards (.97% purity) were obtained from Dr.
ing chemical contaminants. In 1976, Hashemy-Tonkabony et al. Ehrenstorfer GmbH. SPE columns with graphite packing
[4] conducted a survey to monitor the level of chlorinated pesti- (Carbograph; Extract-Clean, 500 mg) were obtained from
cide residues at various stages of vegetable oil processing from Alltech Inc.
seven different oil refineries in Iran. The survey revealed and
confirmed that the commercial deodorizing process was effective 2.2 Standard solution
in removing pesticide residues. A similar survey carried out by
Chaudry et al. [5] in Central Illinois, USA, also reached the same A stock solution was prepared by dissolving 10.0 mg ace-
conclusion and they found that all the pesticide residues tested phate, methamidophos and monocrotophos (each) in 100 mL
accumulated in the deodorization condensate. iso-octane/acetone (9 : 1 vol/vol) to give a final concentration
Since then, a few other published reports that mainly of 100.0 mg/mL. The stock solution was used for sample
discussed the improvement of analytical methods for the spiking and working standard preparations.
analysis of organochlorine pesticide residues in the products
of vegetable oil refining [6, 7] have appeared in various 2.3 Samples
journals. Recently, Pages-Xatart-Pares et al. [8] reported
their findings in their study on the influence of the refining Crude palm oil was obtained from a nearby palm oil mill. The oil
process on pesticide residue removal. In their pilot plant- obtained was subjected to a screening process to ensure that it is
scale study, the effects of both the physical and chemical free from the pesticide residues under study. This oil, termed as
refining process were tested on crude sunflower and rape- blank crude palm oil, was then used for the investigation.
seed oil spiked with pesticides (dichlorvos, malathion, pir- Fortified samples (0.1 and 1.0 mg/g) were prepared by
imiphos-methyl and fenitrothion). The pesticide residues spiking 0.1 and 1.0 mL of stock solution, respectively, into
were monitored at each stage of the refining process and 100 g blank crude palm oil. The spiked crude palm oil was
they concluded that each pesticide studied showed a behav- stirred at 50 7C for 30 min to ensure that a homogeneous
ior particular to its chemical and physical characteristics, mixture was obtained.
especially its chemical reactivity and thermal stability. How-
ever, the recommendations from their studies were to use 2.4 Cleanup and gas chromatographic analysis
high deodorizing temperatures and a high quantity of strip-
ping steam. Another comprehensive review regarding the All samples were analyzed using an in-house-developed
studies of the fate of pesticide residues during refining can method [11] with a recovery range of 85–109%, relative
be found in the IUPAC technical report by Holland et al. standard deviation of less than 11% and a method limit of
[9]. detection of 0.01 mg/g.
From the literature review, it can be seen that the In brief, a 5-g portion of spiked crude palm oil sample was
majority of works carried out to study the behaviors of pes- first weighed into a 50-mL polyethylene screw cap centrifuge
ticide residues during the refining process were with soft tube.
oils. These soft oils possess physical and chemical properties A volume of 10 mL of acetonitrile was then added to the
different from palm oil which is a semi-solid oil at room spiked samples (melted at 60 7C) and shaken for 5 min, after
temperature. To date, the only report on the fate of pesticide which the entire mixture was allowed to stand in a refrigerator
residues in palm oil was from Halimah et al. [10]. Their (4–6 7C) for a minimum of 10 min to allow phase separation
study showed that the degumming-bleaching process of crude palm oil and acetonitrile.
removed 70% of the residue in palm oil spiked with 1.0 mg/g A 2-mL aliquot of the acetonitrile extract (1 g crude palm
chlorpyrifos and the remaining amount was completely oil equivalent) was then loaded onto an SPE column (pre-
removed after deodorization. washed with 10 mL acetonitrile) and allowed to flow through

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Eur. J. Lipid Sci. Technol. 2009, 111, 593–598 OP residues during refining of crude palm oil 595

under gravity; acetonitrile (13 mL) followed by 2 mL metha- the acceleration of volatilization activity at the higher pro-
nol was used to elute the analytes (1.5–2 mL/min). The com- cessing temperature of 105 7C. However, no further study
bined eluate was then evaporated to dryness with a nitrogen has been carried out to study the respective role of the
evaporator and the residue was re-diluted with 1 mL acetone bleaching earth and temperature in the removal of pesticide
before gas chromatographic (GC) analysis. If any dilution was residues.
required, an appropriate dilution was made from this final The role of the deodorizing step in the removal of pesticide
volume. residues in this study was not seen, as all the pesticide residues
were completely removed during the bleaching process.
2.5 Laboratory-scale refining However, the possible role of the deodorizing process in the
removal of pesticide residues was clearly seen in another
The refining experiments were carried out according to experiment using crude palm oil spiked with a higher dose of
the method described in reference [12]. The method used acephate, 5 mg/g (not reported; Fig. 3). In this high-dose
was identical to that of the commercial refining process in experiment, only 90–95% of acephate was removed by the
terms of processing conditions and materials used. The bleaching process and the remaining 5–10% of acephate was
refining experiment was intended to simulate the com- removed by the deodorization process, suggesting that deo-
mercial refining process. In brief, 100 g crude palm oil dorization is important in removing residual pesticides carried
(spiked with 10 or 100 mg pesticides) was weighed into a over from the bleaching process. It has been mentioned that
250-mL three-neck round-bottom flask. The crude palm volatilization is also the main route for the removal of pesticide
oil was heated to 90 7C under nitrogen, with continuous residues and deodorization is carried out at volatilization
agitation. A volume of 0.5 mL citric acid (20% wt/vol) was temperature.
then added and the degumming process was allowed to
continue for 10 min. Next, for the bleaching step, 1 g
commercial bleaching earth (SBE Gold) was added to the 4 Conclusions
oil mixture and the whole mixture was heated to 105 7C
for 15 min. Immediately after this, the mixture was filtered Processing conditions have a great effect on the removal of
through a Whatman No. 1 filter paper with the aid of pesticide residues from spiked crude palm oil. In this study,
vacuum to minimize the hot oil exposure time to air. The results from laboratory-scale palm oil refining experiments
filtered oil was then subjected to deodorization under suggested that the commercial refining process is capable of
vacuum for 20 min at 260 7C. During the refining process, effectively removing three organophosphorus pesticide resi-
samples were collected after each step (degumming, dues, namely, acephate, methamidophos and monocrotophos
bleaching and deodorization) for residue analysis. Three (virtually up to 100% removal after the full refining process),
replicate trials were carried out for crude palm oil spiked from crude palm oil.
with methamidophos, acephate and monocrotophos at the Thus, consumers as well as refineries need not be con-
concentrations of 0.1 and 1.0 mg/g. cerned about these pesticide residues in refined palm oil al-
though these three organophosphorus pesticides are used in
oil palm plantations.
3 Results and discussion In addition, the net effect of processing on pesticide resi-
dues in refined palm oil should be taken into consideration
Figures 1 and 2 show the levels of pesticide residues at differ- when pesticide-related regulations are drawn up with regard to
ent stages of the refining process for crude palm oil spiked at food safety (e.g. MRL).
0.1 and 1.0 mg/g, respectively. It is obvious that degumming is
not an effective process for the removal of the three organo-
phosphorus pesticides from spiked crude palm oil (25–100% Acknowledgments
of spiked pesticides remained in the crude palm oil after
degumming). This is attributed to the fact that, during the The authors would like to thank the management of the MPOB
degumming process, the only route for the removal of pesti- for permission to publish this article and to all supporting staff
cides in crude palm oil is through the volatilization process, who were involved throughout the completion of this study.
and the temperature used during the degumming process Gratitude is also extended to Dr. Tan Yew Ai for her suggestions
(90 7C) is not high enough to volatilize the pesticides com- and guidance in carrying out the experiments as well as writing
pletely from the crude palm oil. of this article.
During the bleaching process at the temperature of
105 7C, the addition of bleaching earth effectively removed
the pesticide residues to a level below the method quantifi- Conflict of interest statement
cation limit. This could be due to the absorption of pesticide
residues on the surface of the bleaching earth coupled with The authors have declared no conflict of interest.

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596 C. B. Yeoh and C. L. Chong Eur. J. Lipid Sci. Technol. 2009, 111, 593–598

Figure 1. Residues of (a) methamidophos,

(b) acephate and (c) monocrotophos at differ-
ent stages of palm oil refining, % (0.1 mg/g
spiking level).

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Eur. J. Lipid Sci. Technol. 2009, 111, 593–598 OP residues during refining of crude palm oil 597

Figure 2. Residues of (a) methamido-

phos, (b) acephate and (c) mono-
crotophos at different stages of palm oil
refining, % (1.0 mg/g spiking level).

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598 C. B. Yeoh and C. L. Chong Eur. J. Lipid Sci. Technol. 2009, 111, 593–598

Figure 3. Removal of acephate in fully refined palm oil (extreme case: 5 mg/g spiking level).

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