• 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.

• 대한화학회지
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• 제40권8호
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• pp.557-564
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• 1996

각기 다른 양이온$(Na^+, Li^+, K^+, NH_4^+)$을 구조이성질체인 일련의 합성 올리고당에 첨가함으로써, 양이온의 결합정도가 올리고당의 연결위치에 따라 달라지리라는 것을 FAB CAD MS/MS(Fast Atom Bombardment Collision Activated Dissociation Mass Spectrometry/ Mass Spectrometry)를 이용하여 안정성 측면에서 연구하였다. 알카리 양이온화된(cationized) 올리고당들은 양성자화(protonated)된 형태보다 더 큰 안정성을 나타내어 -40 eV의 충돌 에너지 수중에서 연결부위와 작용기들이 분절된 스펙트럼형태를 보인 반면, 양성자화된 올리고당은 -10 eV에서 에서 같은 정도의 분절을 보였다. 첨가된 양이온 중, 칼륨 양이온이 첨가된 올리고당이 다른 것에 비해 안정화되어 triple quadrupole기종의 허용에너지 조건하에서는 분절이온을 생성하지 않는다. 다른 양이온화된 올리고당들은 충돌기체인 argon의 압력이 0.8mTorr 상태에서 양이온의 종류에 따라$Nap^+>Li^+>NH_4^+$순으로 안정성을 나타내었다. 올리고당과 결합하는 금속양이온들은 질소를 포함하고 있는 아미노당에 위치하며, 이는 아미노당을 포함하는 분절이온마다 각 양이온에 해당하는 만큼의 질량이 이동된 분절피크들이 나타나는 것으로 설명 가능하다. 또한 양이온화된 것이 양성자화된 것보다 더 큰 안정성을 지니는 이유는 금속 양이온과 아미노당의 N-acetyl 작용기 및 fucose의 산소 사이에 크라운에테르 형태의 결합을 형성하는 것으로 설명할 수 있다. 이 때 참여한 금속 양이온의 크기 및 연결위치-이성질 부분의 구조적 형태에 따라 결합정도의 차이를 보여 각기 다른 분절스펙트럼의 형태를 나타내며 안정성면에서도 1-6>1-4>1-3 연결위치의 순으로 나타났다.

• Journal of Ginseng Research
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• 제44권1호
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• pp.105-114
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• 2020

Background: Exploring the pharmacokinetic (PK) changes of various active components of single herbs and their combinations is necessary to elucidate the compatibility mechanism. However, the lack of chemical standards and low concentrations of multiple active ingredients in the biological matrix restrict PK studies. Methods: A putative multiple reaction monitoring strategy based on liquid chromatography coupled with mass spectrometry (LC-MS) was developed to extend the PK scopes of quantification without resorting to the use of chemical standards. First, the compounds studied, including components with available reference standard (ARS) and components lacking reference standard (LRS), were preclassified to several groups according to their chemical structures. Herb decoctions were then subjected to ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry analysis with appropriate collision energy (CE) in MS² mode. Finally, multiple reaction monitoring transitions transformed from MS² of ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry were used for ultrahigh-performance liquid chromatography coupled with triple quadrupole mass spectrometry to obtain the mass responses of LRS components. LRS components quantification was further performed by developing an assistive group-dependent semiquantitative method. Results: The developed method was exemplified by the comparative PK process of single herbs Radix Ginseng (RG), Radix Polygala (RP), and their combinations (RG-RP). Significant changes in PK parameters were observed before and after combination. Conclusion: Results indicated that Traditional Chinese Medicine combinations can produce synergistic effects and diminish possible toxic effects, thereby reflecting the advantages of compatibility. The proposed strategy can solve the quantitative problem of LRS and extend the scopes of PK studies.

• Natural Product Sciences
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• 제24권3호
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• pp.206-212
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• 2018

Xanthii Fructus has been traditionally used for the treatment of rhinitis, rheumatoid arthritis, and eczema. In this study, a high-performance liquid chromatography-photodiode array (HPLC-PDA) method was developed and then used for the simultaneous analysis of eight phenylpropanoids in Xanthii Fructus. The analytical column used for this separation was a $SunFire^{TM}$ $C_{18}$ column, maintained at $40^{\circ}C$. The mobile phase used was 1.0% acetic acid in distilled water and 1.0% acetic acid in acetonitrile with gradient elution. For identify of each component, the mass spectrometer (MS) was used a Waters triple quadrupole mass spectrometer requipped with electrospray ionization (ESI) source. The HPLC-PDA method showed good linearity: correlation coefficients were ${\geq}0.9996$. The limits of detection and quantification of the eight compounds were 0.02 - 0.04 and $0.06-0.14{\mu}g/mL$, respectively. The extraction recoveries ranged from 97.51 to 108.67%. The relative standard deviation values of intra- and inter-day precision were 0.06 - 1.55 and 0.09 - 1.68%, respectively. The validated HPLC-PDA method was applied to simultaneously analyse the amounts of eight phenlypropanoids in Xanthii Fructus.

• Toxicological Research
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• 제29권3호
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• pp.203-209
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• 2013

A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of ${\varepsilon}$-acetamidocaproic acid (AACA), the primary metabolite of zinc acexamate (ZAC), in rat plasma by using normetanephrine as an internal standard. Sample preparation involved protein precipitation using methanol. Separation was achieved on a Gemini-NX $C_{18}$ column ($150mm{\times}2.0mm$, i.d., 3 ${\mu}m$ particle size) using a mixture of 0.1% formic acid-water : acetonitrile (80 : 20, v/v) as the mobile phase at a flow rate of 200 ${\mu}l/min$. Quantification was performed on a triple quadrupole mass spectrometer employing electrospray ionization and operating in multiple reaction monitoring (MRM) and positive ion mode. The total chromatographic run time was 4.0 min, and the calibration curves of AACA were linear over the concentration range of 20~5000 ng/ml in rat plasma. The coefficient of variation and relative error at four QC levels were ranged from 1.0% to 5.8% and from -8.4% to 6.6%, respectively. The present method was successfully applied for estimating the pharmacokinetic parameters of AACA following intravenous or oral administration of ZAC to rats.

• 한국연초학회지
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• 제33권1호
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• pp.21-27
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• 2011

Amadori compounds(1-deoxy-1-amino-2-ketoses) are important precursors of color, flavor and aroma produced in foods. Amadori compounds occur naturally in tobacco. The contribution of amadori compounds to smoke quality has been (of) interest because of their roles of the Maillard reaction in the leaf chemistry. The amounts of these compound in tobacco are affected by the processes of aging, drying and storage conditions. In this study, eight compounds were chemically synthesized because amadori compounds (have not been sold commercially these days.) were not available for obtaining commercially. The aim of this study was to develop the analytical method of amadori compounds in tobacco leaf by the liquid chromatography mass spectrometry using triple quadrupole analyzer(LC-MS/MS). This method was simple, rapid, selective and sensitive, and eight amadori compounds were simultaneously and quantitatively analyzed within 20 minutes. This method showed excellent accuracy and precision. Recovery rates of amadori compounds ranged from 86% to 102%, with relative standard deviation(RSD) ranged from 2.6% to 5.9%. This method was applied to analysis of amadori compounds contents of tobacco leaves in different varieties. Furthermore, it was expected that the method could be extended to the analysis of other amadori compounds.

• Bulletin of the Korean Chemical Society
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• 제32권2호
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• pp.524-530
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• 2011

In the present study, we demonstrate that SRM LC-MS/MS method developed by Luo et al. (ref. 10) can be successfully applied to the quantitative analysis of intracellular metabolites in E. coli that are collected at the exponential and stationary growth phases. A focus is given on measuring the changes in the concentrations of intracellular metabolites in batch cultures, which were induced during both the dynamically changing exponential and stationary growth phases. The following intracellular metabolites are quantified in the exponential and stationary phases of E. coli growth, using the SRM mode of a triple quadrupole mass spectrometer: glucose-1-phosphate, fructose-1,6-bisphosphate, phosphoenolpyruvate, pyruvate, acetyl-coenzyme A, 6-phosphogluconate, ribulose-5-phosphate, xylulose-5-phosphate, erythrose-4-phosphate. The determined intracellular metabolite concentration profiles are shown to be in a good agreement with the growth profiles of E. coli, which clearly indicates that SRM LC-MS/MS can be successfully used for following the metabolite changes induced at different growth stages.

• Bulletin of the Korean Chemical Society
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• 제35권1호
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• pp.197-203
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• 2014

A rapid, accurate and precise method for the determination of 22 amino acids in Isatidis Radix by Hydrophilic Interaction Ultra-High-Performance Liquid Chromatography Coupled with Triple-Quadrupole Mass Spectrometry (HILIC-UPLC-MS/MS) was established. Chromatographic separation was carried out on a Acquity UPLC BEH Amide column ($2.1mm{\times}100mm$, $1.7{\mu}m$) with gradient elution of acetonitrile (containing 0.05% formic acid and 2 mM ammonium formate) and water (containing 0.15% formic acid and 10 mM ammonium formate) at a flow rate of 0.4 mL/min; Waters Xevo$^{TM}$ TQ worked in multiple reaction monitoring mode. All components were separated in 17 min. All calibration curves were linear ($R^2$ > 0.991) over the tested ranges. The limits of detection (LOD) and limits of quantitation (LOQ) for these compounds were 0.21-79.55 and 0.72-294.23 ng/mL, respectively. The average recoveries were in the range of 93.75-104.16% with RSD value less than 6.56%. Therefore, this method could be an alternative assay for the determination of 22 amino acids in Isatidis Radix due to its rapidness, sensitivity, less sample and solvent consumption.

• Parasites, Hosts and Diseases
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• 제56권6호
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• pp.553-558
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• 2018

Pathogenic Acanthamoeba spp. cause granulomatous amoebic encephalitis and keratitis. Acanthamoeba keratitis (AK) is a rare but serious ocular infection that can result in permanent visual impairment or blindness. However, pathogenic factors of AK remain unclear and treatment for AK is arduous. Expression levels of proteins secreted into extracellular space were compared between A. castellanii pathogenic (ACP) and non-pathogenic strains. Two-dimensional polyacrylamide gel electrophoresis revealed 123 differentially expressed proteins, including 34 increased proteins, 7 qualitative increased proteins, 65 decreased proteins, and 17 qualitative decreased proteins in ACP strain. Twenty protein spots with greater than 5-fold increase in ACP strain were analyzed by liquid chromatography triple quadrupole mass spectrometry. These proteins showed similarity each to inosine-uridine preferring nucleoside hydrolase, carboxylesterase, oxygen-dependent choline dehydrogenase, periplasmic-binding protein proteinases and hypothetical proteins. These proteins expressed higher in ACP may provide some information to understand pathogenicity of Acanthamoeba.

• Mass Spectrometry Letters
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• 제9권4호
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• pp.95-99
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• 2018

An innovative, simple, and rapid assay method based on liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was developed and validated for the simultaneous determination of eight statin drugs in human urine. A simple sample clean-up procedure using the "dilute and shoot" (DAS) approach enabled a fast and reliable analysis. The influence of the dilution factor was investigated to ensure detectability and reduce the matrix effect. Chromatographic separation was performed on a Phenomenex Kinetex C18 column ($50{\times}3.0mm$ i.d., $2.6{\mu}m$) using an elution gradient of mobile phase A composed of 0.1% acetic acid, and mobile phase B composed of acetonitrile, at a flow rate of 0.35 mL/min. Quantitation was performed on a triple quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) mode using electrospray ionization in positive ion mode. The total chromatographic run time was 15 min. The method was validated for selectivity, sensitivity, recovery, linearity, accuracy, precision, and stability. The present method was successfully applied to the analysis of Rosuvastatin in urine samples after oral administration to healthy human subjects.