• 농업생명과학연구
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• 제46권6호
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• pp.165-171
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• 2012

As a preceding study for investigating the influence of sound wave stimulus on Arabidopsis thaliana metabolomics, the polar secondary metabolomes of the plant were determined using high performance liquid chromatography coupled with tandem mass spectrometry. A total of 10 polar secondary metabolomes were characterized and quantified. Among them, 4 metabolomes, p-coumaroylagmatine isomer (7 and 8), p-coumaroylagmatine isomer (9 and 10) were identified in the plant for the first time. The validation was conducted in terms of linearity, recovery, precision, limit of detection (LOD) and limit of quantification (LOQ). The validated method was applied to the simultaneous quantification of the 10 polar secondary metabolomes.

• Environmental Engineering Research
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• 제23권3호
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• pp.309-315
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• 2018

Chlortetracycline (CTC) is one of the most important compounds in antibiotic production, and its distribution has been widely investigated due to health and ecological concerns. This study presents systematic approach to optimize the high-performance liquid chromatography-tandem mass spectrometry for analyzing CTC in a multiple reaction monitoring mode ($479{\rightarrow}462m/z$). One-factor-at-a-time (OFAT) test with response surface analysis (RSA) was used as optimization strategy. In OFAT tests, the fragmentor voltage, collision energy, and ratio of acetonitrile in the mobile phase were selected as major factors for RSA. The experimental conditions were determined using a composite in cube design (CCD) to maximize the peak area. As a result, the partial cubic model precisely predicted the peak area response with high statistical significance. In the model, the (solvent composition) and (collision $energy^2$) terms were statistically significant at the 0.1 ${\alpha}$-level, while the two-way interactions of the independent variables were negligible. By analyzing the model equation, the optimum conditions were derived as 114.9 V, 15.7 eV, and 70.9% for the fragmentor voltage, collision energy, and solvent composition, respectively. The RSA, coupled with the CCD, offered a comprehensive understanding of the peak area that responds to changes in experimental conditions.

• Journal of Ginseng Research
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• 제42권2호
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• pp.149-157
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• 2018

Background: Panax notoginseng leaves (PNL) exhibit extensive activities, but few analytical methods have been established to exclusively determine the dammarane triterpene saponins in PNL. Methods: Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and HPLC-UV methods were developed for the qualitative and quantitative analysis of ginsenosides in PNL, respectively. Results: Extraction conditions, including solvents and extraction methods, were optimized, which showed that ginsenosides Rc and Rb3, the main components of PNL, are transformed to notoginsenosides Fe and Fd, respectively, in the presence of water, by removing a glucose residue from position C-3 via possible enzymatic hydrolysis. A total of 57 saponins were identified in the methanolic extract of PNL by UPLC/Q-TOF MS. Among them, 19 components were unambiguously characterized by their reference substances. Additionally, seven saponins of PNL-ginsenosides Rb1, Rc, Rb2, and Rb3, and notoginsenosides Fc, Fe, and Fd-were quantified using the HPLC-UV method after extraction with methanol. The separation of analytes, particularly the separation of notoginsenoside Fc and ginsenoside Rc, was achieved on a Zorbax ODS C8 column at a temperature of $35^{\circ}C$. This developed HPLC-UV method provides an adequate linearity ($r^2$ > 0.999), repeatability (relative standard deviation, RSD < 2.98%), and inter- and intraday variations (RSD < 4.40%) with recovery (98.7-106.1%) of seven saponins concerned. This validated method was also conducted to determine seven components in 10 batches of PNL. Conclusion: These findings are beneficial to the quality control of PNL and its relevant products.

• Journal of Ginseng Research
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• 제48권4호
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• pp.366-372
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• 2024

Background: The biological activity and pharmacological effects of rare ginsenosides have been proven to be superior to those of the major ginsenosides, but they are rarely found in ginseng. Methods: Ginsenoside Rb1 was chemically transformed with the involvement of methanol molecules by a synthesized heterogeneous catalyst 12-HPW@MeSi, which was obtained by the immobilization of 12-phosphotungstic acid on a mesoporous silica framework. High-performance liquid chromatography coupled with mass spectrometry was used to identify the transformation products. Results: A total of 18 transformation products were obtained and identified. Methanol was found to be involved in the formation of 8 products formed by the addition of methanol molecules to the C-24 (25), C-20 (21) or C-20 (22) double bonds of the aglycone. The transformation pathways of ginsenoside Rb1 involved deglycosylation, addition, elimination, cycloaddition, and epimerization reactions. These pathways could be elucidated in terms of the stability of the generated carbenium ion. In addition, 12-HPW@MeSi was able to maintain a 60.5% conversion rate of Rb1 after 5 cycles. Conclusion: Tandem and high-resolution mass spectrometry analysis allowed rapid and accurate identification of the transformation products through the characteristic fragment ions and neutral loss. Rare ginsenosides with methoxyl groups grafted at the C-25 and C-20 positions were obtained for the first time by chemical transformation using the composite catalyst 12-HPW@MeSi, which also enabled cyclic heterogeneous transformation and facile centrifugal separation of ginsenosides. This work provides an efficient and recyclable strategy for the preparation of rare ginsenosides with the involvement of organic molecules.

• 한국환경보건학회지
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• 제40권2호
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• pp.114-126
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• 2014

Objectives: We aimed to develop a measurement method of five metabolites of trichloroethylene (TCE) in a concurrent biological sample, e.g., trichloroacetic acid (TCA), dichloroacetic acid (DCA), S-(1,2-dichlorovinyl) glutathione (DCVG), S-(1,2-dichlorovinyl)-L-cysteine (DCVC), and N-Acetyl-S-(1,2-dichlorovinyl)-L-cysteine (NAcDCVC) and to validate the method before application to pharmacokinetic study. Methods: TCE metabolites were simultaneously analyzed using high performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS/MS) with as little as 50 ${\mu}L$ of serum and urine. DCA, TCA and NAcDCVC were extracted with diethyl ether, while DCVC and DCVG were extracted by solid phase extraction. This method was validated according to the guidelines for bioanalytical method validation of the Korean National Institute of Toxicological Research. Then, we determined the five metabolites in five strains of mice at 24 hr after exposure to 1 g TCE /kg body weight. Results: The limits of detection for the five metabolites in biological samples ranged from 0.001 to 0.076 nmol/mL, which is comparable to or better than those previously reported. Most calibration curves showed good linearity ($R^2=0.99$), and between-batch variation was less than 20% expressing acceptable robustness and reproducibility. Using this method, we found TCA and DCA were detected in all test mice at 24 hr after the oral administration while NAcDCVC and DCVC were detected in some strains, which showed strain-dependent metabolism of TCE. Conclusions: The present method could provide robust and accurate measurements of major key metabolites of TCE in biological media, which allowed concurrent analysis of TCE metabolism for limited amounts of biospecimens.

• Journal of Ginseng Research
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• 제40권4호
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• pp.382-394
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• 2016

Background: Ginsenosides are the characteristic and principal components which manifest a variety of the biological and pharmacological activities of the roots and rhizomes of Panax ginseng (GRR). This study was carried out to qualitatively and quantitatively determine the ginsenosides in the cultivated and forest GRR. Methods: A rapid and sensitive ultra-high-performance liquid chromatography coupled with diode-array detector and quadrupole/time of flight tandem mass spectrometry (UPLC-DAD-QTOF-MS/MS) was applied to the qualitative analysis of ginsenosides and a 4000 QTRAP triple quadrupole tandem mass spectrometer (HPLC-ESI-MS) was applied to quantitative analysis of 19 ginsenosides. Results: In the qualitative analysis, all ingredients were separated in 10 min. A total of 131 ginsenosides were detected in cultivated and forest GRR. The method for the quantitative determination was validated for linearity, precision, and limits of detection and quantification. 19 representative ginsenosides were quantitated. The total content of all 19 ginsenosides in the forest GRR were much higher than those in the cultivated GRR, and were increased with the growing ages. Conclusion: This newly developed analysis method could be applied to the quality assessment of GRR as well as the distinction between cultivated and forest GRR.

• Food Science and Biotechnology
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• 제18권4호
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• pp.1001-1012
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• 2009

A total of 49 phenolic compounds were identified from the aged red wines made from Cabernet Gernischt (Vitis vinifera L. cv.) grapes, a Chinese characteristic variety, including 13 anthocyanins, 4 pryanocyanins, 4 flavan-3-ol monomers, 6 flavan-3-ol polymers, 7 flavonols, 6 hydroxybenzoic acids, 5 hydroxycinnamic acids, 3 stilbenes, and 1 polymeric pigment. Evolution of these compounds was investigated in wines aged 1 to 13 months. Variance analysis showed that the levels of most phenolics existed significant difference in between wines aged 3 and 9 months. Cluster analysis indicated that 2 groups could be distinguished, one corresponding to wines aged 1 to 3 months and the other to wines aged 4 to 13 months. It was thus suggested that there were 2 key-stages for the development of fine wine quality, at the aged 3 and 9 months, respectively. This work would provide some helpful information for quality control in wine production.

• Journal of Ginseng Research
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• 제42권3호
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• pp.270-276
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• 2018

Background: Notoginsenoside Ft1 is a promising potential candidate for cardiovascular and cancer disease therapy owing to its positive pharmacological activities. However, the yield of Ft1 is ultralow utilizing reported methods. Herein, an acid hydrolyzing strategy was implemented in the acquirement of rare notoginsenoside Ft1. Methods: Chemical profiles were identified by ultraperformance liquid chromatography coupled with quadruple-time-of-flight and electrospray ionization mass spectrometry (UPLC-Q/TOF-ESI-MS). The acid hydrolyzing dynamic changes of chemical compositions and the possible transformation pathways of saponins were monitored by ultrahigh-performance LC coupled with tandem MS (UHPLC-MS/ MS). Results and conclusion: Notoginsenoside Ft1 was epimerized from notoginsenoside ST4, which was generated through cleaving the carbohydrate side chains at C-20 of notoginsenosides Fa and Fc, and vinaginsenoside R7, and further converted to other compounds via hydroxylation at C-25 or hydrolysis of the carbohydrate side chains at C-3 under the acid conditions. High temperature contributed to the hydroxylation reaction at C-25 and 25% acetic acid concentration was conducive to the preparation of notoginsenoside Ft1. C-20 epimers of notoginsenoside Ft1 and ST4 were successfully separated utilizing solvent method of acetic acid solution. The theoretical preparation yield rate of notoginsenoside Ft1 was about 1.8%, which would be beneficial to further study on its bioactivities and clinical application.

• 한국식품위생안전성학회지
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• 제33권5호
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• pp.354-360
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• 2018

견과류 중에 있는 트리코테센계 곰팡이독소 오염도를 조사하기 위하여 LC-MS/MS를 이용한 정확성과 신뢰성을 동시에 확보할 수 있는 분석방법을 개발하였다. 견과류 중 트리코테센계 곰팡이 독소는 QuEChERS 추출 및 EMR-Lipid-dSPE 정제과정을 통하여 분석에 사용되었다. 검량선 작성을 위하여 트리코테센계 곰팡이독소 10종에 대하여 $2.00{\sim}75.00{\mu}g/kg$의 범위로 혼합표준용액을 제조하여 실험하였으며, 상관계수는 모두 0.998 이상으로 높은 직선성을 나타내었다. 분석방법의 검출한계는 $0.41{\sim}3.57{\mu}g/kg$로 나타났으며, 정량한계는 $1.23{\sim}10.82{\mu}g/kg$로 나타났다. 또한 트리코테센계 곰팡이 독소 10종에 대하여 각각 저, 중, 고 3가지 농도로 처리하여 회수율 실험을 수행한 결과 81.84~96.87%로 나타났다. 확립된 분석법으로 견과류 중 땅콩을 대상으로 오염도를 조사한 결과 1종에서 deoxynivalenol이 검출되었다. 이러한 결과를 바탕으로 확립된 시험법은 견과류 중 트리코테센계 곰팡이 독소 분석에 적합함을 확인할 수 있었으며 견과류 중 트리코테센계 곰팡이 독소 검출 가능성을 확인한바 보다 다양한 종류의 견과류에 대한 모니터링 조사가 필요한 것으로 판단된다.

• 한국식품위생안전성학회지
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• 제32권5호
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• pp.389-395
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• 2017

루이보스티에서 5종의 phenolic acid (gallic acid, chlorogenic acid, caffeic acid, p-coumaric acid, trans-ferulic acid)와 9종의 flavonoid (procyanidin b1, aspalathin, rutin, vitexin, hyperoside, isoquercitrin, luteolin, quercetin, chrysoeriol)를 UPLC-MSMS를 이용하여 동시 분석하였다. 14종 페놀릭류를 동시 분석하기 위하여 기기조건과 유효성을 검증하였고 확립된 분석방법을 이용하여 시중에 유통중인 루이보스티 30건을 채취하여 페놀릭류를 분석하였다. 루이보스티 1 g 혹은 1티백에 뜨거운 물 100 mL을 가하여 3분, 6분, 30분이 경과 후 그리고 차가운 물 ($25-30^{\circ}C$)에 30분 우려낸 루이보스티의 페놀릭류 함량을 구하였다. 루이보스티에서 전체 실험대상 페놀릭류 중 rutin과 aspalathin이 가장 많이 추출되어 나왔으며 각각 물질의 함량은 제품별로 달랐다. 페놀릭류 성분의 추출효율은 14종 페놀릭류의 총합 기준으로 뜨거운 물 30분 > 6분 > 3분 > 차가운 물 30분 순으로 높았다.