• Natural Product Sciences
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• 제22권2호
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• pp.93-101
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• 2016

For efficient quality control of the Samryeongbaekchul-san decoction, a powerful and accurate an ultra-performance liquid chromatography (UPLC) coupled with electrospray ionization (ESI) tandem mass spectrometry (MS) method was developed for quantitative analysis of the thirteen constituents: allantoin (1), spinosin (2), liquiritin (3), ginsenoside Rg1 (4), liquiritigenin (5), platycodin D2 (6), platycodin D (7), ginsenoside Rb1 (8), glycyrrhizin (9), 6-gingerol (10), atractylenolide III (11), atractylenolide II (12), and atractylenolide I (13). Separation of the compounds 1 - 13 was performed on a UPLC BEH $C_{18}$ column ($2.1{\times}100mm$, $1.7{\mu}m$) at a column temperature of $40^{\circ}C$ with a gradient solvent system of 0.1% (v/v) formic acid aqueous-acetonitrile. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}L$. Calibration curves of all compounds were showed good linearity with values of the correlation coefficient ${\geq}0.9920$ within the test ranges. The values of limits of detection and quantification for all analytes were 0.04 - 4.53 ng/mL and 0.13 - 13.60 ng/mL. The result of an experiment, compounds 2, 6, 12, and 13 were not detected while compounds 1, 3 - 5, and 7 - 11 were detected with 1,570.42, 5,239.85, 299.35, 318.88, 562.27, 340.87, 12,253.69, 73.80, and $115.01{\mu}g/g$, respectively.

• 생약학회지
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• 제48권4호
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• pp.320-328
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• 2017

Yongdamsagan-tang has been used to treat the urinary disorders, acute- and chronic-urethritis, and cystitis in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was established for simultaneous analysis of the 20 bioactive marker compounds, geniposidic acid, chlorogenic acid, geniposide, liquiritin apioside, acteoside, calceolarioside B, liquiritin, nodakenin, baicalin, liquiritigenin, wogonoside, baicalein, glycyrrhizin, wogonin, glycyrrhizin, wogonin, saikosaponin A, decursin, decursinol angelate, alisol B, alisol B acetate, and pachymic acid in traditional herbal formula, Yongdamsagan-tang. Chromatographic separations of all marker compounds were conducted using a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was performed using a Waters ACQUITY TQD LC-MS/MS coupled with an electrospray ionization source in the positive and negative modes. The flow rate was 0.3 mL/min and injection volume was $2.0{\mu}L$. The correlation coefficient of 20 marker compounds in the test ranges was 0.9943-1.0000. The limits of detection and quantification values of the all marker components were 0.11-6.66 and 0.34-19.99 ng/mL, respectively. As a result of the analysis using the optimized LC-ESI-MS/MS method, three compounds, geniposidic acid (from Plantaginis Semen), alisol B (from Alismatis Rhizoma), and pachymic acid (from Poria Sclerotium), were not detected in this sample. While the amounts of the 17 compounds except for the geniposidic acid, alisol B, and pachymic acid were $0.04-548.13{\mu}g/g$ in Yongdamsagan-tang sample. Among these compounds, baicalin, bioactive marker compound of Scutellariae Radix, was detected at the highest amount as a $548.13{\mu}g/g$.

• 생약학회지
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• 제47권1호
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• pp.92-101
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• 2016

In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was established for simultaneous determination of the 25 marker components, including chlorogenic acid, gallic acid, oxypaeoniflorin, homogentisic acid, methyl gallate, caffeic acid, 3,4-dihydroxybenzaldehyde, paeoniflorin, albiflorin, liquiritin, nodakenin, ferulic acid, ginsenoside Rg1, liquiritigenin, coumarin, cinnamic acid, benzoylpaeoniflorin, ginsenoside Rb1, cinnamaldehyde, paeonol, glycyrrhizin, 6-gingerol, evodiamine, rutecarpine, and spicatoside A in traditional Korean formula, Onkyung-tang. All analytes were separated on a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was carried out using a Waters ACQUITY TQD LC-MS/MS coupled with an electrospray ionization (ESI) source in the positive and negative modes. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}L$, respectively. The correlation coefficient of all analytes in the test ranges was greater than 0.98. The limits of detection and quantification values of the 25 marker compounds were in the ranges 0.03-19.43 and 0.09-58.29 ng/mL, respectively. As a result, methyl gallate, 3,4-dihydroxybenzaldehyde, evodiamine, and rutecarpine were not detected in this sample and the concentrations of the 21 compounds except for the above 4 compounds were $33.09-3,496.32{\mu}g/g$ in Onkyung-tang decoction. Among these compounds, paeonol was detected at the highest amount as a $3,496.32{\mu}g/g$.

• Journal of Ginseng Research
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• 제38권3호
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• pp.187-193
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• 2014

Background: White ginseng (Panax ginseng Meyer) is commonly distributed as a health food in food markets. However, there is no practical method for distinguishing Korean white ginseng (KWG) from Chinese white ginseng (CWG), except for relying on the traceability system in the market. Methods: Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry combined with orthogonal partial least squares discrimination analysis (OPLS-DA) was employed to discriminate between KWG and CWG. Results: The origins of white ginsengs in two test sets ($1.0{\mu}L$ and $0.2{\mu}L$ injections) could be successfully discriminated by the OPLS-DA analysis. From OPLS-DA S-plots, KWG exhibited tentative markers derived from ginsenoside Rf and notoginsenoside R3 isomer, whereas CWG exhibited tentative markers derived from ginsenoside Ro and chikusetsusaponin Iva. Conclusion: Results suggest that ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry coupled with OPLS-DA is an efficient tool for identifying the difference between the geographical origins of white ginsengs.

• Journal of Ginseng Research
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• 제44권2호
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• pp.205-214
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• 2020

Background: This article aims to compare and analyze the contents of ginsenosides in ginseng of different plant ages from different localities in China. Methods: In this study, 77 fresh ginseng samples aged 2-4 years were collected from 13 different cultivation regions in China. The content of eight ginsenosides (Rg₃, Rc, Rg₁, Rf, Rb₂, Rb₁, Re, and Rd) was determined using rapid resolution liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry (RRLC-Q-TOF MS/MS) to comparatively evaluate the influences of cultivation region and age. Results: Ginsenoside contents differed significantly depending on age and cultivation region. The contents of ginsenosides Re, Rc, Rg₁, Rg₃, and Rf increased with cultivation age, whereas that of ginsenoside Rb₁ peaked in the third year of cultivation. Moreover, the highest ginsenoside content was obtained from Changbai (19.36 mg/g) whereas the lowest content was obtained from Jidong (12.05 mg/g). Ginseng from Jilin Province contained greater total ginsenosides and was richer in ginsenoside Re than ginseng of the same age group in Heilongjiang and Liaoning provinces, where Rb₁ and Rg₁ contents were relatively high. Conclusion: In this study, RRLC-Q-TOF MS/MS was used to analyze ginsenoside contents in 77 ginseng samples aged 2-4 years from different cultivation regions. These patterns of variation in ginsenoside content, which depend on harvesting location and age, could be useful for interested parties to choose ginseng products according to their needs.

• 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.

• 한국식품위생안전성학회지
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• 제33권1호
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• pp.58-64
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• 2018

본 연구는 소의 가식부위(근육, 신장, 간장, 지방) 중에서 세팔렉신을 효과적으로 정량분석하기 위한 LC-MS/MS법을 확립하고 이를 검증하기 위해 수행되었다. 확립된 LC-MS/MS에 대해 특이성, 검출한계, 정량한계, 정확도 및 정밀도에 대한 검증을 통하여 유효성을 확인하였다. 표준 용액을 이용하여 검량성을 작성한 결과, $r^2$ > 0.999 이상의 직선성을 나타내었으며, 세팔렉신에 대한 검출한계와 정량한계는 각각 2~10과 $6{\sim}30{\mu}g/kg$으로 나타났다. 또한, 회수율은 83.9~106.8%로 나타났으며, 상대표준편차는 2.3~14.8%로 나타나 정확성이 우수하였다. 이는 식품의약품안전처의 잔류동물용의약품 분석법에서 제시한 기준에 모두 적합한 수준이었다. 따라서 본 연구를 통해 개발된 LC-MS/MS법은 향후 소의 가식부위 중 세팔렉신을 분석하는데 효과적으로 활용될 수 있을 것으로 사료된다.

• 한국식품위생안전성학회지
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• 제32권5호
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• pp.418-423
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• 2017

본 연구는 우유 중에서 덱사메타손을 효과적으로 정량분석하기 위한 LC-MS/MS법을 확립하고 이를 검증하기 위해 수행되었다. 확립된 LC-MS/MS에 대해 특이성, 검출한계, 정량한계, 정확도 및 정밀도에 대한 검증을 통하여 유효성을 확인하였다. 표준용액을 이용하여 검량성을 작성한 결과, $r^2$ > 0.999 이상의 직선성을 확인하였고, 덱사메타손에 대한 검출한계와 정량한계는 각각 0.15와 0.5 ng/mL이었다. 또한, 회수율은 98.9-109.6%로 나타났으며, 상대표준편차는 1.7-4.4%로 나타나 정확성이 우수하였으며, 이는 식품의약품안전처의 잔류동물용의약품 분석법에서 제시한 기준에 모두 적합한 수준이었다. 따라서 본 연구를 통해 개발된 LC-MS/MS법은 향후 우유 중 덱사메타손을 분석하는데 효과적으로 활용될 수 있을 것으로 사료된다.

• Journal of Ginseng Research
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• 제39권4호
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• pp.338-344
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• 2015

Background: Ginseng (the roots of Panax ginseng Meyer) is a well-known traditional Oriental medicine and is now widely used as a health food. It contains several types of ginsenosides, which are considered the major active medicinal components of ginseng. It has recently been reported that the qualitative and quantitative properties of ginsenosides found in ginseng may differ, depending on cultivation regions, ages, species, and so on. Therefore, it is necessary to study these variations with respect to cultivation ages and regions. Methods: In this study, 3-6-yr-old roots of P. ginseng were collected from three different cultivation regions. The contents of five ginsenosides (Rb1, Rd, Rc, Re, and Rgl) were measured by rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry. The Kruskal-Wallis Rank sum test and multiple t test were used for comparative analysis of the data to evaluate the dynamic changes in the accumulation of these ginsenosides affected by cultivation regions and ages. Results: The content and composition of ginsenosides varied significantly among specimens collected from different cultivation regions and having different cultivation ages. For all samples, the content of Rg1 and Re ginsenosides increases with age and this rate of increase is different for each sample. The contents of Rb1, Rc, and Rd varied with cultivation ages in samples from different cultivation regions; especially, Rb1 from a 6-yr-old root showed approximately twofold variation among the samples from three cultivation regions. Furthermore, the content of Rb1 highly correlated with that of Rd (r = 0.89 across all locations and ages). Conclusion: In our study, only the contents of ginsenosides Rg1 and Re were affected by the root age. Ginsenosides Rb1, Rc, and Rd varied widely with ages in samples from different cultivation regions.

• Journal of Pharmaceutical Investigation
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• 제41권4호
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• pp.255-262
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• 2011

Method using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was developed and validated for the determination of pregabalin in plasma samples. Acquisition was performed by monitoring the transitions: m/z 160.1${\rightarrow}$142.2 for pregabalin and m/z 423.2${\rightarrow}$207.1 for losartan (as an internal standard). After cold acetonitrileinduced protein precipitation of the plasma samples, separation was performed with C18 column by isocratic mobile phase consisted of 10 mM ammonium acetate and acetonitrile (15:85, v/v). Results were linear over the concentration ranged from 0.1 to $10{\mu}g$/mL and the correlation coefficients (r) were $\geq0.99$. Intra- and inter-day precisions were $\leq6.02$ and $\leq11.04%$, respectively, and intra- and inter-day accuracies were 96.60-101.09 and 98.10-102.60%, respectively. This validated method was successfully applied to a bioequivalence study of two formulations of pregabalin, Daewoong pregabalin capsule (Daewoong Pharm. Co., Ltd.) and Lyrica$^{(R)}$ capsule (Pfizer Korea Ltd.) in twenty eight healthy Korean volunteers. The subjects received a single oral dose of each formulation (150 mg as pregabalin) in a randomized $2{\times}2$ crossover study and plasma samples were obtained from each subject at predetermined time intervals. Then, the pharmacokinetic parameters ($AUC_{0-t}$, $C_{max}$ and $T_{max}$) were calculated and statistically analyzed to assess the differences between two formulations. The 90% confidence intervals for the log-transformed data were acceptable range of log 0.8-log 1.25 (e.g., log 1.0048-log 1.0692 for AUC0-t, log 0.9142-log 1.0421 for $C_{max}$). Thus, $AUC_{0-t}$ and $C_{max}$ met the criteria of the Korea Food and Drug Administration (KFDA) for bioequivalence test indicating that Daewoong pregabalin capsule was bioequivalent to Lyrica$^{(R)}$ capsule.