• Korean Journal of Environmental Agriculture
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• v.29 no.4
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• pp.388-395
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• 2010

In order to decrease level of an organophosphorus fungicide, tolclofos-methyl, from in situ ginseng cultivating soil, we isolated a tolclofos-methyl degrading bacteria from ginseng cultivating soil samples. The bacterial strain removed tolclofos-methyl around 95% after 3 days incubation with complete liquid media. The strain was identified as Sphingomonas sp. by 16S rDNA sequence comparison, and designated as Sphingomonas sp. 224. Through the GC-MS analysis, Sphingomonas sp. 224 was proposed to have an initiative degradation pathway generating the metabolite such as 2,6-dichloro-4-methyl phenol compound from tolclofos-methyl. In addition, Sphingomonas sp. 224 was confirmed representing the effective degrading capability to tolclofosmethyl in situ soil.

• Korean Journal of Environmental Agriculture
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• v.29 no.4
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• pp.396-401
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• 2010

Tolclofos-methyl is one of the most widely used organophosphorous pesticides in control of soil-borne diseases in ginseng field. In Korea, residues of tolclofosmethyl in ginseng and cultivation soil is quite often detecting. The objective of this study was to know the possibility for the accelerated degradation of tolclofos-methyl by various chemical treatment under soil slurry condition. The degradation of tolclofos-methyl was accelerated by zerovalent metals treatment in soil slurry. The degradation rate of tolclofos-methyl was found to be at higher zerovalent zinc than unannealed zerovalent and annealed zerovalent iron. The effect of different sizes of zerovalent iron on tolclofos-methyl degradation was showed that the smaller size of zerovalent iron, the greater the degradation rate. In aqueous solution of pH 4.0 below the degradation rate of tolclofos-methyl was very high. Under this experimental condition, tolclofos-methyl degradation was the greatest at 2% (w/v) of ZVI under 0.1 N of HCl in 24 hours, the degradation rate was 94.4%. By testing various chemicals, it was found that $Fe_2(SO_4)_3$ as iron source showed better for degrading tolclofos-methyl in $H_2O_2$ 500 mM treatment and sodium sulfite also showed the degradable possibility tolclofos-methyl in soil slurry.

• The Korean Journal of Pesticide Science
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• v.18 no.3
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• pp.130-140
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• 2014

Recently, some of the previous studies reported that tolclofos-methyl is still exist in ginseng cultivated soil, even though it is has been banned for ginseng. Therefore, the current study was aimed to examine the levels of absorption and translocation of tolclofos-methyl from ginseng cultivated soil to ginseng root and leaf stem for the period of 1 year. For this study, ginseng plants were transplanted in pots and treated with $5.0mg\;kg^{-1}$ of tolclofos-methyl (50% WP). At the end of each interval periods (every three months) the samples (soil, roots and leaf stems) were collected and analyzed the absorption and translocation levels of tolclofos-methyl using gas chromatography and mass spectrometry (GC-MS). The limit of quantitation of tolclofos-methyl was found to be $0.02mg\;kg^{-1}$ and 70.0~120.0% recovery was obtained with coefficient of variation of less than 10% regardless of sample types. In this study, a considerable amount of translocation of tolclofos-methyl residues were found in soil (4.28 to $0.06mg\;kg^{-1}$), root (7.09 to $1.54mg\;kg^{-1}$) and leaf stem (0.79 to $0.69mg\;kg^{-1}$). The results show that the tolclofos-methyl was absorbted and translocated from ginseng cultivated soil to ginseng root and ginseng leaf stem and found to be decreased time-coursely. Secondly, we were also analyzed soil, root and leaf stems samples from Hongcheon, Cheorwon, Punggi and Geumsan by GC-MS/MS (172 pesticides), LC-MS/MS (74 pesticides). In this study, 43 different pesticides were detected ($0.01{\sim}7.56mg\;kg^{-1}$) in soil, root and leaf stem. Further, tolclofos-methyl was detected 4 times separately in root sample alone which is less ($0.01{\sim}0.05mg\;kg^{-1}$) than their maximum residual limit (MRL) in ginseng. Consequently, the results from both studies indicate the residues of tolclofos-methyl found in ginseng cultivated soil and ginseng ensuring their safety level. Moreover, long-term evaluations are needed in order to protect the soil as well as ginseng free from tolclofos-methyl residues.

• Journal of Ginseng Research
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• v.13 no.1
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• pp.114-118
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• 1989

Tolclofos-methyl applied as seed dipping at 1,000 ppm for 3 hrs before sowing and soil drenching at the rate of 300 g ai./10 a in the middle of April protected emerging seedlings of Panax ginseng from damping-off caused by Rhiiutonia solani(AG2-1) in Yangjik Soil artificially infested with the pathogen. Germination rates with tolclofos-methyl, pencycuron, and control were 53.7%, 45.8%, and 7.5%, respectively, while the rate of the seeds at non-infested soil was 62.6%. The effectiveness of Tolclofos-methyl against the pathogen in the soil lasted upto 32 days in vitro. However, the transpiratio of ginseng seedlings increased greatly with chemical treatment, showing 0.02, 0.12, and 0.24 m1/cm2 leaf area/day at 0, 1,2, and 4 ppm a.i. of the fungicide, respectively.

• Korean Journal of Environmental Agriculture
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• v.28 no.4
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• pp.409-411
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• 2009

Phytochelatins (PCs) are small-sized peptides synthesized by PC synthase (PCS) using glutathione (GSH) as a substrate, and they play an important role in the detoxification of toxic heavy metals in plants, fission yeast, and other living organisms. Recently, it has been suggested that PCS is also involved in degradation of some xenobiotics including monobromobimane. PCS cleaves the Gly residue from GSH-xenobiotics conjugates resulting in ${\gamma}$-Glu-Cys-xenobiotics, and this is to degraded further. Therefore, our research is focus on whether PCS is also involved in degradation of tolclofos-methyl, an important pesticide which has been used in ginseng cultivated areas. Heterologous expression of Arabidopsis PCS confers tolerance to tolclofos-methyl in yeast. Furthermore, PCS-deficient Cad1-3 Arabidopsis mutant showed high sensitivity to tolclofos-methyl compared with wild-type plants. These results imply that PCS is involved in degradation of tolclofos-methyl as other xenobiotics.

• Korean Journal of Environmental Agriculture
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• v.26 no.4
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• pp.337-342
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• 2007

To obtain the data on the risk assessment of residue levels of organophosphorus and organochlorine pesticides in fresh ginseng and red ginseng extract, the residual pesticides in samples are surveyed with GC-NPD and GC-ECD for quantitative analysis and GC-MSD for qualitative analysis. The residual organophosphorus pesticides, such as diazinon, pyrimethanil, tolclofos-methyl, metalaxyl, diethofencarb, parathion, cyprodinil, tolylfluanid and kresoxim-methyl, are not detected in fresh ginseng from Punggi. The residual organophosphorus pesticides except tolclofos-methyl are not detected in fresh ginseng from Sangju. Average amount of tolclofos-methyl in fresh ginseng from Sangju are $0.054{\pm}0.008\;mg/kg$, representing $18{\pm}2%$ of MRL of 0.03 mg/kg on fresh ginseng in Korea. The residual organochlorine pesticides, such as BHC isomer, DDT isomer, aldrin, azoxystrobin, captan, cypermethrin, deltamethrin, dieldrin, difenoconazole, endosulfan-sulfate, endrin, fenhexamid, quintozene, ${\alpha}$-endosulfan and ${\beta}$-endosulfan, are not detected in fresh ginseng from Punggi and Sangju. The residual organophosphorus and organochlorine pesticides in red ginseng extract from Punggi and Sangju are not detected.

• Bulletin of the Korean Chemical Society
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• v.24 no.3
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• pp.334-338
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• 2003

A simple synthetic method for haptens of organophosphorus (OP) pesticides with a spacer arm (aminocarboxylic acid) attached at the pesticide thiophosphate group was developed and was applied to the synthesis of haptens for the OP fungicide tolclofos-methyl. Using the haptens, a selective enzyme-linked immunosorbent assay (ELISA) for tolclofos-methyl was developed. One of the haptens was coupled to BSA to use as an immunogen. Rabbits were immunized with this conjugate to obtain polyclonal antibodies to tolclofos-methyl. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the serum with highest specificity, an antigen-coated ELISA was developed, which showed an $IC_{50}$ of 160 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivity with other OP pesticides. An antibody-coated ELISA was also developed, which showed an $IC_{50}$ of 410 ng/mL with a detection limit of 130 ng/mL.

• The Korean Journal of Pesticide Science
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• v.16 no.3
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• pp.217-221
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• 2012

This study was carried out to evaluate residual characteristics of tolclofos-methyl in ginseng and elucidate the reason for its high detection rate from fresh ginseng selling at markets. Seeds of ginseng were sowed after seed dressing with tolclofos-methyl and after a year of growth, the young seedlings were transplanted to field. They were then harvested annually until three-years of growth and the pesticide residue was analyzed in them. LOD and LOQ of the pesticide were 0.001 and 0.003 mg/kg, respectively. Recovery test was carried out to validate the analytical method for tolclofos-methyl in ginseng. The ginseng seedlings were fortified with the test pesticide at the level of LOQ, ten times of LOQ and maximum residue concentration of tolclofos-methyl. Its recovery ranged from 77.37 to 100.16%. Residual concentration of tolclofos-methyl in ginseng seedlings just before transplanting and two-year-old ginseng were from 7.58 to 8.05 and from 6.46 to 6.79 mg/kg, respectively. In case of three-year-old ginseng, it was found to be from 4.18 to 4.35 mg/kg. As a result of annual pesticide residue analysis, concentration of the pesticide was found to decrease time-coursely in ginseng. This may be due to decomposition and increasing of fresh weight of the ginseng during the cultivation periods of three years.

• Journal of Food Hygiene and Safety
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• v.14 no.4
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• pp.365-371
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• 1999

Microwave-assisted extraction (MAE) is known as a more environmental-friendly process with economic advantages in terms of less time, less solvent, less energy and less waste than the current reflux method that is time-consuming. MAE was applied to extract three pesticides (quintozene, tolclofos-methyl and procymidone) from ginseng in a pesticide residue analysis and eliminate pesticide contaminants from ginseng prior to production of the ginseng extracts. The optimal conditions for pesticide extraction from ginseng were 50% in power (150 watts), 2 minutes, 20 mL hexane and 2.0 g sample. The results of optimal conditions were compared with those of Soxhlet method (7 hours, 150 mL hexane). The recoveries between two methods were almost same while time and amount of solvent used in MAE were significantly decreased in comparing with those in the Soxhlet methods. When the 45% ethyl alcohol as a extraction solvent was applied to eliminate pesticides from ginseng prior to the production of the ginseng extracts, it was found that 100% quintozene, 69％ tolclofos-methyl and 83% procymidone were eliminated from ginseng. The data showed that MAE may be used to eliminate pesticide contaminants in ginseng prior to making the ginseng extracts.

• 한국약용작물학회:학술대회논문집
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• 2008.11a
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• pp.491-492
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• 2008