J. Sep. Sci.

2007, 30, 1953 – 1963

H-R. Jeon et al.


Hyang-Rang Jeon1, 2* Mostafa Abd El-Aty1, 3, 4* Soon-Kil Cho2 Jeong-Heui Choi1 Kil-Yong Kim5 Ro-Dong Park5 Jae-Han Shim1

Original Paper Multiresidue analysis of four pesticide residues in water dropwort (Oenanthe javanica) via pressurized liquid extraction, supercritical fluid extraction, and liquid–liquid extraction and gas chromatographic determination
The primary objective of this study was to simultaneously analyze the residues of the most commonly used pesticides, chlorpyrifos-methyl, endosulfan, EPN, and iprodione in the water dropwort, via accelerated solvent extraction (ASE), supercritical fluid extraction (SFE), and conventional solvent extraction (LLE) techniques. Residue levels were determined using GC with electron-capture detection (GC-ECD). The confirmation of pesticide identity was performed by GC-MS in a selected ion-monitoring (SIM) mode. In none of the ASE and SFE techniques were the extraction conditions optimized. Rather, the experimental variables were predicated on the author's experience. The ECD response for all pesticides was linear in the studied range of concentrations of 0.005 – 5.0 ppm, with correlation coefficients in excess of 0.9991. At each of the two studied fortification levels, the pesticides yielded recoveries in excess of 72% with RSDs between 1 and 19%. The LODs were achieved at a range of levels from 0.001 to 0.063 ppm, depending on the pesticide utilized. The LOQs, which ranged from 0.003 to 0.188 ppm, were lower than the maximum residue limits (MRLs) authorized by the Korean Food and Drug Administration (KFDA). All of the methods were applied successfully to the determination of pesticide residues in the real samples. It could, therefore, be concluded that any of the techniques utilized in this investigation might prove successful, given that the applied extraction conditions are wisely chosen.
Keywords: Dropwort / Multiresidue / Pressurized solvent extraction / Simultaneous / Supercritical fluid extraction / Received: December 22, 2006; revised: March 12, 2007; accepted: March 14, 2007 DOI 10.1002/jssc.200600548

Natural Products Chemistry Laboratory, Institute of Agricultural Science and Technology, College of Agriculture and Life Science, Chonnam National University, Buk-Ku, Gwangju, Republic of Korea 2 National Agricultural Products Quality Management Service, Products Safety Inspection Laboratory, Gwangsan-Gu, Gwangju, Republic of Korea 3 Department of Pharmacology, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt 4 Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, Konkuk University, KwangjinGu, Seoul, Republic of Korea 5 Glucosamine Saccharide Materials Laboratory (NRL), Division of Applied Bioscience and Biotechnology, Chonnam National University, Buk-Ku, Gwangju, Republic of Korea

1 Introduction
Pesticides can potentially exert adverse effects on vegetables, fruits, animal resources, and human health [1]. Thus, the determination of pesticide residues in foods and other environmental components/commodities including fruits, vegetables, total diet, soil, and water has become an increasingly essential requirement for
Correspondence: Professor Jae-Han Shim, Natural Products Chemistry Laboratory, Institute of Agricultural Science and Technology, Chonnam National University, 300 Yong-Bong Dong, Buk-Ku, Gwangju 500-757, Republic of Korea. E-mail: jhshim@chonnam.ac.kr Fax: +82-62-530-0219 Abbreviations: ASE, accelerated solvent extraction; ECD, electron-capture detector; LLE, liquid – liquid extraction; MRLs, maximum residue limits; SFE, supercritical fluid extraction

consumers, producers, and authorities for food quality control. Although classical liquid – liquid extraction (LLE) or solid – liquid extraction methods are often tedious, time consuming, and require prodigious amounts of organic solvents, they remain the most commonly utilized extraction or preconcentration techniques, either alone or combined with others in analyses of pesticides [2]. Several concerns about the hazards associated with most of the solvents used, coupled with the costs and environmental dangers inherent to waste solvent disposal, resulted in the development of more efficient environmentally friendly techniques for the rapid analyticalscale extraction of solid matrices, including the supercritical fluid extraction (SFE) technique [3]. SFE has gained an increasing amount of attention as a potential
* Both authors contributed equally to this work.


2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

have been frequently employed for this purpose. because the diffusion of analytes from the sample to the solvent can be considerably augmented via the reduction of particle size. SFE/HPLC). the stems and leaves are employed as salad components or as a seasoning in soups and stews in the Republic of Korea. Organophosphates constitute the primary insecticides used with dropwort. Sci. and encourages analyte diffusion to the matrix surface. Also. the aforementioned methods were applied to the analysis of real samples. such as diatomaceous earth or cellulose. however. CO2 as a nonpolar substance is capable of dissolving both nonpolar and moderately polar compounds. due to its supercritical fluid properties. and carries with it the advantage of low cost and nonspecific instrumentation demands. 2. The key factors in the extraction process associated with pesticide solubility in supercritical fluid include pesticide desorption from the matrix surface and the diffusion of the desorbed pesticide into the bulk solvent. 0. 0. preventing them from adsorption on the active sites of the sample matrix [4]. and endosulfan is the principal organochlorine insecticide used that exerts a selective effect against arthropod attacks. Drying agents.05. 0. The multiresidue determination of pesticide residues in various matrices using ASE [6 – 8] and SFE [9 – 18] has been previously reported. Upon the application of SFE. Pretreatment normally involves either the sieving or grinding of the sample. Temperature also facilitates the breakdown of analyte – matrix bonds. acetone. Germany). The modifiers increase the solubility of analytes. Germany). and EPN (95.5%) were provided by Dr. 0.0625. Unless stated otherwise. Jeon et al.125. The drying of the sample is also important. and 0.8%). methanol. This step is particularly important when nonpolar solvents are to be employed in extraction. 2007. The most important advantages of SFE include: considerable reduction of the volume of used solvent. 0.025. the primary objective of the present work was to determine the conditions for the simultaneous determination of chlorpyrifos-methyl. Working standard solutions containing 0. 0. Individual stock solutions were prepared via the weighing of appropriate amounts of active ingredients in pesticide-residue-grade acetone at a concentration of 1000 lg/ mL. 0. The compound chemical structures are shown in Table 1. time. and 5.1954 H-R.025. 0.1 Analytical standards and miscellaneous materials Pesticide reference standards. prior to loading into the extraction cell.com i 2007 WILEY-VCH Verlag GmbH & Co. all other chemicals were of analytical grade. KGaA. and endosulfan-sulfate). It is a member of the Umbelliferae family (along with coriander.1 ppm (chlorpyrifos-methyl and EPN).25. as any moisture may diminish the extraction efficiency. 2 Experimental 2. the working standards solutions were also stored at a temperature of –248C.05. extraction is performed using supercritical CO2. Sep. 0. Weinheim .625. there have as yet been no reports regarding dedicated techniques for the analysis of pesticide residues in the dropwort. caraway. b-endosulfan (99%). This is generally achieved by the optimization of CO2 pressure and the temperature of the extraction cell. 1. small required sample size. The advantages of ASE include minimized environmental concerns. A mixture of CO2 and modifiers (polar organic solvents) can be used to extract polar substances. n-hexane. fennel.01. Accelerated solvent extraction (ASE) or pressurized liquid extraction (PLE) is another new extraction technique. and anhydrous sodium sulfate were purchased from Merck KGaA (Darmstadt. iprodione (99%). LC-Florisil SPE cartridge for pesticide purification.jss-journal. as it is known to be a more suitable method for the routine analysis of pesticide traces. endosulfan-sulfate (98. The resultant concentration was then corrected for the stated purity. which allows for faster extraction than is possible via classical methods. chlorpyrifos-methyl (99%). Stock solutions of individual compounds were stored in aluminum/Teflon-lined capped vials at –248C. and a discussion of results is presented. the sample is frequently pretreated in some fashion.5.0125. These solutions were utilized in the construction of calibration curves and the preparation of fortified samples. including higher diffusivity and low viscosity [3].05.0 ppm (iprodione) were also prepared in acetone. which increases the solvent's ability to wet the matrix and to solubilize the target analytes. 1953 – 1963 replacement for the conventional LLE method. www.025. and 0. and cumin). PLE is a sample preparation method that couples elevated temperature and pressure with liquid solvents in order to effect a fast and efficient extraction of the analytes from solid matrices [5]. labor. ease of automation. GCMS was used to confirm the pesticides analyzed by GC. Water dropwort (Oenanthe javanica) is a perennial herb with a distinctive aroma and taste. high degree of selectivity. iprodione and EPN in dropwort.5 ppm (a-endosulfan. 0.25. and 0. shortened extraction time. The ASE technique is also well suited to the automation and online coupling of extraction and separation. Thus. using ASE and SFE. a-endosulfan (98. b-endosulfan. The application of higher temperatures implies a reduction in solvent viscosity. and the possibility of online coupling with separation and determination techniques (SFE/GC. The LLE technique was also employed. endosulfan. and is cultivated in marshy areas of Asia and Australia. 0. J.5%). high degree of purity and small extract volume. and its metabolite. Ehrenstorfer GmbH (Augsburg. and also minimized exposure of laboratory technicians to toxic chemicals.005. After this point. 30.

2. The total extract volume (30 mL) was then transferred into 100 mL pear-shaped flasks and was concentrated to approximately 3 mL using a Büchi Rotavapor R-114 at 408C. and divided into four subsamples. concentrated in a rotary evaporator under reduced pressure at 408C.jss-journal. A mixture of acetone/n-hexane (1:4 v/v) was utilized as an extraction solvent in all cases [8]. Tokyo. Gwangju. The extract was then transferred into 100 mL pear-shaped flasks. Japan) was then introduced into the cell. After extraction. the samples were placed into polyethylene bags and transported to the laboratory. CA. and subjected to GC-ECD analysis. Republic of Korea) that specializes in the sale of organic crops. On the Dionex ASE 200. the empty space above the mixture was filled with florisil (2 g). dichloromethane. Japan). it was determined that there were no detectable levels of target analytes in the samples prior to spiking via solvent extraction method. 30. thoroughly mixed. 2007. which is located in the southern region of the Korean peninsula and the Jollanamdo area. The solvent was collected in 60 mL vials with Teflon septa. followed by the mixture. The residue was dissolved in 2 mL of acetone. The samples were extracted using a Jasco SFE PU980 dual pump (Tokyo. 2 g of sea sand standard (Junsei Chemical.J. Sunnyville. Thereafter. where they were chopped. and n-hexane for at least 24 h in each solvent. between January 2004 and September 2005.4 SFE extraction Control samples were purchased from a special market (Lotte Mart. Sep.3 ASE extraction Sample extractions were conducted using an automated Dionex ASE 200 system (Dionex. then dried under a gentle stream of pure nitrogen. Chem tube hydromatrix (4 g). after 5 min of equilibration. Finally. action. CA. The flow rate was 3 mL/ min. USA) (4 g) was added to the beaker as a drying agent and the contents were mixed with a spatula. Palo Alto. The remaining volume was evaporated to dryness at 558C. Approximately 3 g of dropwort samples were extracted using a 10 mL stainless steel vessel. USA) at a i 2007 WILEY-VCH Verlag GmbH & Co. Carbon dioxide modified with 30 vol% methanol was used [19 – 22]. Two cellulose microfilters were positioned at each end of a Dionex 33 mL cell. After collection. In our laboratory. Nine real samples (approximately 2 – 3 kg) were obtained from Jeju Island.com 2. 2. Twenty grams were randomly selected and monitored in triplicate for pesticide residues via GC-ECD. Weinheim . After the fortification of any spikes with the appropriate amounts of fortification solution. the cell was flushed with solvent (60%) and purged with nitrogen. The residues were then dissolved in 2 mL of acetone and stored at 48C until injection at room temperature on a gas chromatograph equipped with an electron-capture detector (ECD). Sci. Chem tube hydromatrix (Varian. using a gentle nitrogen stream. A 20 g portion of the www.5 Liquid – liquid extraction (conventional method) The multiresidue extraction method utilized in the determination of pesticides in dropwort was conducted in accordance with the procedures in the relevant literature [23 – 25]. and the solvent was collected in the same vial. These subsamples were maintained in deep-frozen conditions until analysis. After gently tapping the cell in order to settle the contents.2 Sampling Table 1. approximately 3 g of chopped samples were placed into a 100 mL beaker. the samples were immersed sequentially in acetone. 1953 – 1963 Sample Preparations 1955 2. yielding a free-flowing powder. KGaA. The SFE conditions were as follows: 5 min of static mode and 30 min of dynamic mode at 808C and 300 atm. and sea sand (2 g). the contents of the beakers were permitted to stand at room temperature for 15 – 20 min. Static extraction was conducted for 5 min. with minor modifications. Structures. and classification of the tested pesticides in water dropwort (Oenanthe javanica) Pesticide Chemical structure Action/classification Insecticide acaricide/organophosphorus Chlorpyrifosmethyl Endosulfan Insecticide acaricide/organochlorine Iprodione Fungicide/dicarboximide EPN Insecticide acaricide/organophosphorus temperature of 1008C and a pressure of 1500 atm. each extraction cell containing the same sample was subjected to one additional identical extraction cycle.

The column had been prewashed with 12 mL of n-hexane prior to use. KGaA. The sample was then extracted with 100 mL of acetone and macerated for 5 min at 3000 rpm in a homogenizer (WiseMixTM HG15A. 22.8 LOD and LOQ The LOD and the LOQ are the smallest concentrations from which it is possible to deduce the presence and quantify the analyte. it may swell the matrix and expose small internal cavities. b-endosulfan.6 GC and GC-MS analysis A Hewlett – Packard model 6890 series II gas chromatograph equipped with a 63Ni micro-cell ECD. and concentrated to 2 – 3 mL on a rotary evaporator (Büchi Rotavapor R-114.25 mm id.93.7 Recovery The extraction efficiency (recovery rate) of the tested pesticides was determined in triplicate at two fortification levels in spiked samples of blank water dropwort (i. 19. 1 lL. 24. augments the average recovery of the tested compounds. methanol was utilized as a modifier because it sufficiently increased the solvent power of CO2 for the extraction of several compound classes in the spiked samples. using a gentle nitrogen stream. USA) coated with 5% phenyl and 95% methylpolysiloxane was utilized. maintained for 0 min.66. Under these conditions. 17.1956 H-R. The injection port and interface temperatures were maintained at 230 and 2808C. makeup gas N2 (flow rate: 66. 2508C. The extracts were then purified with 2 g SPE florisil MEGA BE-SI column (45 cm620 mm). dehydrated by passing through a filter containing a bed of anhydrous Na2SO4.59. 26]. Seoul. 20.25 lm.25 mm ID) and 0.1 Extraction using different techniques According to our experience using SFE and the findings of other researchers. endosulfan-sulfate.48. Daihan Scientific.. The residues were dissolved in 2 mL of acetone and stored at 48C until injection at room temperature on a gas chromatograph equipped with an ECD.07. Sep. respectively. Palo Alto. 1953 – 1963 dropwort sample was weighed in a 250 mL homogenizer cup and fortified when required with the standard pesticide solution. SFE conducted at 808C with 5 min static and 30 min dynamic extraction provided us with the extraction efficiency. and EPN in this column and GC conditions were 14. The pesticides were confirmed by their retention time and the identification of target ions with appropriate individual standards. iprodione. Additionally. as previously described. The retention times of chlorpyrifos-methyl. The eluate was evaporated to dryness at 558C. b-endosulfan. It may also effect an increase in the polarity of the supercritical fluid and enhance the partitioning of polar analytes into fluid. and a fused-silica capillary column DP-5 (30 m60.53 min. was then held for 10 min. 30. increased by 28C/min to 2208C. Agilent Technologies. The resulting samples were mixed and allowed to stand for 15 min before extraction. 21. and 22. and the extracts were partitioned using 50 mL of n-hexane in two steps. The recovery rate was calculated by comparing the peak areas for the spiked water dropwort (containing known amounts of pesticides in the range of the calibration curve concentrations) with standard solutions dissolved in an organic solvent and were directly injected into the analytical column.58.999%) as the carrier gas at 1 mL/min.jss-journal. This.3 mL/min). The sample was then loaded and eluted with 20 mL of acetone: n-hexane (1:4 v/v). aendosulfan. J. respectively. An Agilent model 5973N mass spectrometer system was used for the confirmation of the samples and analytes. The oven temperature was held at 1008C for 2 min. respectively. containing none of the pesticides of interest). the retention times of chlorpyrifos-methyl. HP-5MS (30 m60. The LOD and LOQ are equal to the mean of the measured content of representative blank samples plus three times or ten times the SD of the mean. 3008C. and EPN were 14.52 min. 2. The samples were then homogenized for 5 min and then submitted to extraction using the three techniques and GC-ECD analysis. Finally. The homogenate was vacuum filtered using a Büchner funnel fitted with a no. endosulfan-sulfate. 27. 19.99. iprodione. Germany). respectively.25 lm film thickness (Agilent Technologies) coated with 5% phenyl and 95% methylpolysiloxane was employed using helium (99. A fused-silica capillary column. then increased to 2808C at the rate of 58C/min and 2. which compared www. it may compete with polar analytes for active sites in the matrix and displace them into the fluid. The electron-impact energy was 70 eV. an autosampler. with reasonable statistical certainty. 100 mL of a 10% sodium chloride solution was added.com i 2007 WILEY-VCH Verlag GmbH & Co. 2. 6 Whatman filter paper. The filtrate was then quantitatively transferred to a 500 mL separatory funnel. Jeon et al. a-endosulfan. CA. in turn. injection volume. The operating conditions were as follows: initial temperature. allowing for better access of the supercritical fluid into the absorbed analytes [19 – 22. Germany) in a water bath at 408C (Büchi Waterbath B-480. Both phases were combined. detector temperature. operated in a 60:1 split ratio. injector temperature.24. film thickness = 0. respectively. increased to 2008C at the rate of 108C/min and held for 1 min. N2 carrier gas (flow rate: 1 mL/min). maintained for 4 min.15. and 28. Sci. Korea). 2007. e. then increased by 108C/min to 3008C for 6 min. 1308C (2 min). 3 Results and discussion 3. Weinheim . increased by 78C/min to 2008C.

obtained via the injection of standards prepared in the sample extract.025 – 0. e. For ASE. 1953 – 1963 Sample Preparations 1957 Table 2. According to Erney et al. ASE evidenced superior recovery in a range of 82 – 116%. fulfill the requirements of the European Union Guidelines [34]. Cleanup was necessary to remove the polar coextracted substances from the LLE-extracted samples.9991 0.5 0.5 0.jss-journal.1 0.025 – 5.15 1. a temperature of 1008C and a pressure of 1500 atm were applied throughout the experiments [8].9999 0. Extracts analyzed by GC-ECD were found to be largely free of chromatographic interferences (Figs. the extracting solvent should be of a low polarity (such as hexane) Calibration curves were plotted via the injection of standards prepared in solvent.9994 0. pigments.. the external calibration procedures generated a least-square linear regression line with a correlation coefficient of 0. For each analyte.0 0. as it had previously been successfully utilized in our laboratory in the extraction of some of the pesticides involved in the present study from other fruits and vegetables for GC-based analyses [8]. samples with high water content can be dried. [27].025 – 0. as no matrix effects were observed when a comparison was made to curves. The water content of dropwort was 92.01 0.5 5. Table 3.5 0.9991 or higher (Table 2).2 Method performance less favorably to the LLE data. thus large amounts of interfering materials are trapped at the top of the cartridges [33]. 1 and 2). Acetone/hexane was selected as the extraction solvent. they may reduce the selectivity and therefore cause the coextraction of other matrix components [5]. This may have blocked the tube and prevented the passage of CO2 into the tube. In order to assess method accuracy and repeatability. This is consistent with previous reports by Nilsson et al. In all the fortification levels.005 – 0. i 2007 WILEY-VCH Verlag GmbH & Co.9997 to minimize the coextraction of interfering compounds [32].9999 0. ensuring more complete transfer from the injector to the column. In general. [35] organophosphate pesticide recoveries higher than 100% for LLE and ASE are induced by the sample matrix.1 r2 0.0 0. Additionally.15 1. Range and determination coefficient (r2) of the tested pesticides with the GC-ECD method Pesticide Chlorpyrifos-methyl a-Endosulfan b-Endosulfan Endosulfan-sulfate Iprodione EPN Range (ppm) 0. Therefore. g.005 – 0. as expressed in the terms of (mean) RSD.com . KGaA. Additionally. 31] also failed to detect any relationship between pressure and recovery. Although higher temperatures (A 1008C) will increase both solubility and mass transfer.J. followed by LLE. Sep. we utilized the samples in freeze-dried form. some other researchers [4. The findings of the three analytical methods still.025 – 0. however. Florisil retains the more polar interfering materials. which functions as a shield for analyte molecules against loss in hot injectors. This indicated that often reasonable exhaustiveness can be achieved at 808C. The results are presented in Table 3.9999 0. Recoveries of pesticides in dropwort using different analytical methods Pesticide Chlorpyrifos-methyl a-Endosulfan b-Endosulfan Endosulfan-sulfate Iprodione EPN Fortification level (ppm) 0.5 0. A pressure of 1500 atm was selected considering a safety issue with the equipment.01 0. but for dropwort. 3. 30. higher temperatures may be required. extractions were conducted at two levels of fortification.1 0.15 1. The precision of all investigated extraction methods.5 0. Weinheim www. which evidenced recovery rates ranging from 80 to 107%. was favorable.8% [28]. 2007.5 0. The lowest recoveries (72 – 90%) were obtained with the SFE method. Sci. and this water will become ice under supercritical status conditions.1 ASE recovery (M l SD) 116 l 8 113 l 5 89 l 4 95 l 1 90 l 3 101 l 4 82 l 3 106 l 4 89 l 5 100 l 2 101 l 3 114 l 5 RSD (%) 7 5 4 1 4 4 3 3 6 2 3 5 SFE recovery (M l SD) 90 l 2 81 l 12 85 l 2 85 l 1 85 l 10 74 l 12 86 l 16 78 l 11 85 l 10 72 l 11 87 l 3 79 l 14 RSD (%) 2 15 2 1 12 16 19 14 11 16 4 18 LLE recovery (M l SD) 91 l 10 107 l 2 80 l 8 81 l 1 104 l 4 99 l 3 88 l 4 92 l 3 88 l 3 94 l 2 86 l 10 95 l 1 RSD (%) 11 2 10 1 4 3 4 4 4 2 12 1 Results are the mean of three replicates l SD. 30. since drying does not affect the pesticide content [29].

which were determined as the minimum amount of each analyte in the sample matrix necessary for a S/N of at least 3. 3.1 – 5. [36]) (Table 4). the detection limits. 2007. Chlorpyrifosmethyl. which ranged from 0. pressurized – liquid extraction (with 60 mL of solvent consumption at a pressure of 1500 atm and 1008C) over a period of 26 min yielded results comparable to those of LLE (with 265 mL of solvent consumption) conducted for 82 min. A.1958 H-R. The calculated LOQs. as the solvent properties are affected and the mass-transfer efficiency is enhanced. C. and the volume of solvent utilized. D. 30.jss-journal. Sep. B-2. iprodione.com i 2007 WILEY-VCH Verlag GmbH & Co. Weinheim . KGaA. b-endosulfan.025 ppm) in LLE compared to both ASE and SFE methods. a-endosulfan. endosulfan-sulfate. In the present study.188 ppm for all compounds were lower than the maximum residue limits (MRLs) established by the Korean Food and Drug Administration (KFDA) (0. B-3.001 – 0.3 Comparison of extraction techniques The selection of an appropriate extraction technique entails the consideration not only of the recovery but also the cost. Concerning the quantitative results. EPN.003 – 0. On the www. B-1.063 ppm). Higher temperatures and pressures during the extraction profitably influence process efficiency. Jeon et al. were of lower magnitude (0. time of extraction.0 ppm. J.001 – 0. Chromatogram of blank unspiked samples (1) and the pesticide standards dissolved in acetone (2). 1953 – 1963 Figure 1. Sci. which have the same magnitude (0.

Chlorpyrifos-methyl. 30. Sep. Sci. B-3. Weinheim www. B-1. C. Chromatogram of spiked dropwort samples using ASE (1). and LLE (3). endosulfan-sulfate. a-endosulfan. i 2007 WILEY-VCH Verlag GmbH & Co.jss-journal. KGaA.com . SFE (the attenuation was modified to demonstrate the corresponding peaks). b-endosulfan. 1953 – 1963 Sample Preparations 1959 Figure 2. 2007.J. D. iprodione. EPN. B-2. A. (2).

188 0. Sep. Figure 3. Sci.4 Analysis of real samples Table 5 and Fig.001 0. Weinheim . 3.001 0. and LLE. MS was carried out as screening method for the confirmation of all positive results from GC analysis.006 0. B-2.004 SFE LOD (ppm) LOQ (ppm) 0. GC-ECD chromatograms of the detected pesticide in real dropwort samples using ASE. 2007.com i 2007 WILEY-VCH Verlag GmbH & Co.1 1. the value which could reduce the number and magnitude of mistakes.001 0. J. followed by LLE with an RSD ranging between 1 and 12%. A. and maximum residue limits (MRLs. SFE. Chlorpyrifos-methyl.063 0. expressed in terms of a range of RSD between 1 and 7%. analyzed via ASE.075 0. Due to the large probability of false positive results obtained by GC-ECD screening methods.188 0. www.019 0.019 0. As a primary step in the MS method development. KGaA. a-endosulfan. ppm) of the tested pesticides with the GC-ECD method Pesticide LOD (ppm) Chlorpyrifos-methyl a-Endosulfan b-Endosulfan Endosulfan-sulfate Iprodione EPN a) ASE LOQ (ppm) 0.019 0.003 MRLa) (ppm) 0. The total-ion chromatograms (TICs) for all pesticide standards were obtained and all spectral data were stored in the computer library.0 5.025 0.008 0.1960 H-R.006 0. B-3.001 0.001 0. endosulfan-sulfate. B-1. b-endosulfan.008 0. 30.019 0. 1953 – 1963 Table 4.063 0.004 0.0 0. The LOD (ppm).003 0.004 0.1 0. LOQ (ppm).006 0. and LLE methods. ASE evidenced an excellent precision.003 0.019 0.006 0. 3 show the results for the real samples collected in agricultural fields on Jeju Island.006 0. pesticide-working solutions were analyzed by GC-MS scanning in full-scan mode with a scan range from 50 to 550 m/z.001 MRL values determined by KFDA [36]. SFE.006 0.003 0.004 LLE LOD (ppm) LOQ (ppm) 0.008 0. Jeon et al.003 0.jss-journal. confirmatory analysis was needed.019 0. other hand.

The three most representative fragments (a target ion and two qualifiers) and retention time window were selected for each pesticide peak from the TICs stored in the computer. and 207 for a-endosulfan. and were in agreement with those www. KGaA. 241. A. The major selected ions (m/z) were as follows: 286.com i 2007 WILEY-VCH Verlag GmbH & Co. For the confirmation of peak identity GC-MS operating in SIM was applied. Sci. B-1. 229. and 387 for endosulfan-sulfate. 314. and LLE methods. and 79 for chlorpyrifosmethyl. 1953 – 1963 Sample Preparations 1961 Figure 4. and 56 for iprodione. Sep.jss-journal. 237.J. a-endosulfan. 195. SFE. B-3. 169. and 185 for EPN. GC-MS-SIM chromatograms of the detected pesticide in real dropwort samples using ASE. b-endosulfan. Chlorpyrifos-methyl. 195. 125. B-2. 2007. and 207 for b-endosulfan. Weinheim . 272. and 157. 30. 187. endosulfan-sulfate.

. [7] Adou. J. 18.. J. M. Lee. J.01 LLE 0. K. J..08 l 0. 4. Suwon. W. 2005. J. E.01 Residue level (mean l SD) SFE 0.02 reported by other authors for the mass spectra of these compounds [37.01 0. Elsevier. J. [28] Rural Development Administration.. L. H.20 l 0..0 ppm [36]. S. J. R. Y. most notably ASE. J. SFE.... a bit higher quantities of chlorpyrifos-methyl. A 2003. 119. 4153 – 4160.. S. Namienik.. The residue levels of chlorpyrifos-methyl.. 745 – 751. 856. J.. Chromatogr. Shim. Kim. S.. [1] Perez Bendito. Toxicol. B. M.. 1033 – 1039. H..1962 H-R. b-endosulfan. Abd El-Aty. 69.. Sweeney.. 1996. [27] Nilsson. M. J. S. Chromatogr. Heo. [19] Shim. Anal.07 l 0. [6] Yi. 22.. J. C. 303 – 312. J.. and the procedure should be conducted at working temperatures of 1008C for more recent extraction techniques. 88. A 1998. KGaA. J. 1997. R.. The sensitivity of these methods is sufficient to ensure a reliable determination at pesticide levels that were much lower than the respective MRLs established by the KFDA. B.21.. 123 – 135.06 l 0.. Galhiane.6. Kim.. Sci.. a-endosulfan. W. Sep. 4 Concluding remarks In conclusion. R. Fussell. Yamagami.. Jeon et al.. J. Anal.01 0.. Ezzel.01 0. 2007. K.... I. J. [10] Lehotay. [18] Ono. C.. Björklund... Park. Goodall. Korea 2001.. B. J. S. J. J.03 0. Biotechnol. Chromatogr. 615 – 629.01 0. A 2004. M10300000322-06J0000-32210). D. 153 – 159. R. G. 183 – 187.jss-journal. C. Bontoyan. 2004.. [14] Hawthorne. Anal. 38]. Biosci. 821 – 827.21 l 0. [29] Pesticide Residues V. X. Shen. 69 – 84. Van Bodegraven.15 to 0. 1999. Kang. program funded by the Ministry of Science and Technology (no. K.. Cho. B. S. C. J. 1148 – 1166.. Kim. Toyoda. [31] Heemken. [12] Nemoto. Theobald.. A. S. in press. O. 2003. J. A 2005.. M. Hashi. Shim. Shim. 2443 – 2450.. 51. Apon. A. J. H. [16] Rissato. C.. de Kok.. D. and endosulfan-sulfate residues were recorded by both LLE and ASE compared with SFE.. Chim. Bicchi... C. Shen. S. a-endosulfan.. Biochem. S. C. 149.. C. H. Kim.01 0. K. M. G. J.. L. 88. Weinheim . A GC-MS chromatogram for a positive dropwort sample is reported in Fig.. S. Knoll. 464. 2006. J. 2001. S. Anal. Grabanski. S. W. [23] Liu. Rev. Sci.. www. [26] Nerín. S.. B. A. J.07 l 0. Kim. 1997..04 l 0. Y. S. B. Chromatogr. A. B.. S. J. the total sums of endosulfan (aendosulfan. Food Chem.. Park.04 l 0. M. M. J. 892. Martin. R. S. Rapid Commun.. Agric. Y.. Anal. J. 2006. Jones. Sykes. and LLE methods Crop Pesticide ASE Water dropwort Chlorpyrifos-methyl a-Endosulfan b-Endosulfan Endosulfan sulfate 0. 1048. [21] Kang.. A. A. H. Berl. and endosulfan-sulfate were detected in the real dropwort samples by three analytical methods. This work was supported by the Korean Science Foundation (KOSEF) through the National Research Laboratory. J. 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