• 대한한의학방제학회지
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• 제21권1호
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• pp.177-185
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• 2013

Objectives : Leejung-tang (Lizhong-tang) has been used for treatment of gastrointestinal disorders in Korea. In this study, we performed quantification analysis of five marker components, liquiritin, ginsenoside Rb1, ginsenoside Rg1, glycyrrhizin, and 6-gingerol in Leejung-tang using a ultra performance liquid chromatography- electrospray ionization-mass spectrometer (UPLC-ESI-MS). In addition, we evaluated antioxidant activity of Leejung- tang. Methods : The column for separation of five constituents used a UPLC BEH C18 ($100{\times}2.1mm$, $1.7{\mu}m$) maintained at $45^{\circ}C$. The mobile phase consisted of two solvent systems, 0.1% (v/v) formic acid in H2O (A) and CH3CN (B) by gradient flow. The flow rate was 0.3 mL/min with detection at mass spectrometer. The antioxidative activities conduct an experiment on 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of Leejung-tang. Results : Calibration curves of five marker compounds were acquired with r2 values > 0.99. The amount of the five compounds in Leejung-tang were 0.07 - 0.84 mg/g. The concentration required for 50% reduction (RC50) against ABTS radical was 119.02 ug/mL. In addition, the scavenging against DPPH radical of Leejung-tang was 11.4%, 14.5%, 19.8%, 29.6%, and 49.2% at 25 ug/mL, $50{\mu}g/mL$, $100{\mu}g/mL$, $200{\mu}g/mL$, and $400{\mu}g/mL$, respectively. Conclusions : The established LC-MS/MS method will be helpful to improve quality control of Leejung-tang. In addition, Leejung-tang is a potential antioxidant therapeutic agent.

• 생약학회지
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• 제49권3호
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• pp.270-277
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• 2018

Pyungwi-san has been used to treat the digestive system diseases, physconia, nausea, anorexia, and dyspepsia in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was optimized for simultaneous determination of the 14 marker components, spinosin, liquiritirn apioside, liquiritin, narirutin, 6'''-feruloylspinosin, hesperidin, liquiritigenin, glycyrrhizin, 6-gingerol, atractylenolide III, honokiol, atractylenolide II, magnolol, and atractylenolide I in Pyungwi-san extract. All analytes were separated on a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) with maintained at $45^{\circ}C$. The mobile phase consisted of 0.1% (v/v) aqueous formic acid and acetonitrile. The MS conditions were as follows: capillary voltage 3.3 kV, extractor voltage 3.0 V, RF lens voltage 0.3 V, source temperature $120^{\circ}C$, desolvation temperature $300^{\circ}C$, desolvation gas 600 L/h, cone gas 50 L/h and collision gas 0.14 mL/min. The coefficient of determination of 14 analytes was 0.9989-1.0000. The limits of detection and quantification values of the all analytes were 0.04-2.56 and 0.13-7.69 ng/mL, respectively. As a result of the analysis using the established LC-ESI-MS/MS method, the 5 components, spinosin, 6'''-feruloylspinosin, atractylenolide III, II, and I derived from Zizyphi Fructus and Atractylodis Rhizoma, were not detected in this extract. On the other hand, the 9 components except for the 5 components were 4.15-498.87 mg/kg in lyophilized Pyungwi-san extract. Among these components, glycyrrhizin, marker compound of Glycyrrhizae Radix et Rhizoma, was detected the most amount as a 498.87 mg/kg.

• 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 $3,496.32{\mu}g/g$.

• 생약학회지
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• 제47권1호
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• pp.62-72
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• 2016

Banhasasim-tang is a well-known traditional Korean herbal formula and has been used clinically for the treatment of gastric disease, including acute and chronic gastritis, diarrhea and gastric ulcers in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer method was developed for the quantitative determination of the 13 marker constituents, homogentisic acid (1), 3,4-dihydroxybenzaldehyde (2), spinosin (3), liquiritin (4), baicalin (5), ginsenoside Rg1 (6), liquiritigenin (7), wogonoside (8), ginsenoside Rb1 (9), baicalein (10), glycyrrhizin (11), wogonin (12), and 6-gingerol (13) in Banhasasim-tang decoction. Separation of the compounds 1-13 was using an UPLC BEH $C_{18}$ ($100{\times}2.1mm$, $1.7{\mu}m$) column and column oven temperature was maintained at $45^{\circ}C$. The mobile phase consisted of 0.1% (v/v) formic acid in water (A) and acetonitrile (B) by gradient elution. The injection volume and flow rate were $2.0{\mu}L$ and 0.3 mL/min, respectively. Calibration curves of the compounds 1-13 were showed with $r^2$ values ${\geq}0.9908$. The limit of detection and limit of quantification values of the compounds 1-13 were 0.04-1.11 ng/mL and 0.13-3.33 ng/mL, respectively. Among the these compounds, the compounds 1-3 were not detected, while the compounds 4-13 were detected in the ranges of $3.20-107,062.98{\mu}g/g$ in Banhasasim-tang sample.

• 약학회지
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• 제60권2호
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• pp.64-72
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• 2016

Jakyakgamcho-tang is a well-known traditional herbal medicine and has been used for the treatment of mainly pains in oriental medicine. In this study, analytical method for the quantitative determination of the eleven marker components, gallic acid (1), oxypaeoniflorin (2), paeoniflorin (3), albiflorin (4), liquiritin (5), isoliquiritin (6), ononin (7), liquiritigenin (8), benzoylpaeoniflorin (9), paeonol (10), and glycyrrhizin (11) in Jakyakgamcho-tang decoction was performed using an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer. The analytical column for separation of the compounds 1~11 was used an UPLC BEH $C_{18}$ ($100{\times}2.1mm$, $1.7{\mu}m$) column and column oven temperature was maintained at $45^{\circ}C$. The mobile phase consisted of 0.1% (v/v) aqueous formic acid (A) and acetonitrile (B) by gradient elution. The flow rate was 0.3 ml/min and injection volume was $2.0{\mu}l$. Correlation coefficient in the calibration curves of the compounds 1~11 were showed a good linearity with more than 0.99. The limit of detection and limit of quantification values of the compounds 1~13 were detected in the ranges 0.06~18.43 ng/ml and 0.18~58.29 ng/ml, respectively. Among the compounds 1~11, the compounds 10 were not detected in this sample, while the ten compounds, 1~9 and 11, were detected $44.05{\sim}19,289.05{\mu}g/g$ in Jakyakgamcho-tang extract.

• 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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• 제46권2호
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• pp.160-166
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• 2015

Yijung-tang (YJT) is a traditional herbal formula comprising 4 medicinal herbs. In the present study, we performed the simultaneous analysis for three compounds of YJT and examined anti-allergic effects in vitro. The column for separation of three compounds was used Gemini C18 column and maintained at 40$^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems. To evaluate Th2 chemokines, YJT was treated into tumor necrosis factor (TNF)-${\alpha}$ and interferon (IFN)-${\gamma}$-stimulated HaCaT cells, and performed ELISA for thymus and activation regulated chemokine (TARC) and regulated on activation, normal T-cell expressed and secreted (RANTES). To measure Th2 cytokines, YJT was added into primary mouse splenocytes, and performed ELISA for interleukin (IL)-4, 5, 13. Calibration curves were acquired with r² >0.9999. The contents of liquiritin, glycyrrhizin, and 6-gingerol in YJT were 4.50 mg/g, 11.10 mg/g, and 1.33 mg/g, respectively. YJT inhibited production of TARC and RANTES in TNF-${\alpha}$ and IFN-${\gamma}$-treated HaCaT cells. YJT also reduced production of IL-4, 5, and 13 in primary mouse splenocytes. In conclusion, our data will be a valuable information to improve quality control and anti-allergic effects of YJT.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.103-103
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• 2019

Glycyrrhizae radix, commonly known as licorice, is a perennial herb belonging to Leguminosae and also includes various components such as, glycyrrhizin, liquiritin, liquiritigenin and isoliquiritigenin etc. Licorice has been widely used in East Asia as a medicine having pharmacological effects like antioxidants, anti-bacterial, anti-inflammatory, anti-cancer and immune modulatory activities. Among various licorice, Glycyrrhiza (G.) uralensis G. glabra and G. inflata are used for pharmaceutical purposes in Korea. However, cultivation of licorice has some problems such as low quality, low productivity, and early leaf drop. Korea Rural Development Administration developed new cultivars Wongam and Sinwongam, which are improved in cultivation and quality. To register the newly developed cultivar (s) on Ministry of Food and Drug Safety in Korea as a medicine, it is necessary to prove the similarity and difference through the comparative studies between already-registered species and new cultivars. Some fungi and bacteria usually in the human oral cavity and intestines exist as harmless state in human body. Also, the skin and genital infections by fungi can lead to toxic systemic infections and are accompanied by flushing, rashes, burning or painful sensation. The influences of licorice varieties on fungi and bacteria might be an evidence to prove the outstanding effect of newly developed licorice variety. In this study, the antifungal and antibacterial activity was investigated using newly developed licorice varieties Wongam, and Sinwongam against various fungi and bacteria. These results means newly developed licorice could be used as a replacement of already-registered species in terms of antifungal and antibacterial application.

• 생약학회지
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• 제49권4호
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• pp.362-371
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• 2018

Hwanggeumjackyak-tang (HJT) composed of Scatellaria baicalensis Georgi, Paeonia Lactiflora and Glycyrrhizae uralensis Fischer is a traditional Korean herbal medicine widely used for acute enteritis. In order to develop the tablet formulation of HJT, evaluation of the flow properties, thickness, diameter, hardness, friability and disintegration was carried out on four HJT granules according to mixed content of seven additives. Simultaneous analysis used HPLC method was performed of HJT tablet and was determined of the seven marker components; Albiflorin, Paeoniflorin, Liquiritin, Baicalin, Baicalein, Glycyrrhizic acid and Wogonin. The biological activities were examined the effect of HJT on anti-oxidation and pro-inflammation mediated by LPS-stimulation. We confirmed that both of HJT-Decoction (HJT-D) and HJT-Formulation (HJT-F) have the similar contents on total polyphenol and flavonoid and inhibited the secretion of nitro oxide (NO), interleukin $(IL)-1{\beta}$,interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$ and the expression of nitric oxide synthase (iNOS), $IL-1{\beta}$, IL-6, $TNF-{\alpha}$. Therefore, the developed formulation for tablet of HJT would provide chemically and biologically the same effect compared with decoction of HJT.

• 대한본초학회지
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• 제28권4호
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• pp.1-6
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• 2013

Objectives : In other to determine the optimal extraction conditions, the various Bojungikgi-tang (BJIGT) decoctions prepared by different pressure levels and different extraction times were compared and evaluated in terms of the extract yield and the total soluble solid content. Methods : Decoctions were prepared by the pressure levels of 0 (non-pressurized) and 1 $kgf/cm^2$ (pressurized) for 60, 120 and 180 min. The extract yield and the total soluble solids content of decoctions were measured, and the amounts of the reference compounds in decoctions were investigated by the analysis using high performance liquid chromatography. Results : The extract yield and the total soluble solid content were higher in decoctions extracted by the pressurized method than those from decoction with non-pressurized method. The patterns of yield and contents showed a proportional increase to the extraction time. In analysis of the linear regression for four reference compounds such as liquiritin, nodakenin, hesperidin, and glycyrrhizin, the good linearity with the correlation coefficient more than 0.9999 was observed. The highest contents for four reference compounds were observed at 180 min of both the pressurized method and the non-pressurized method. Conclusions : This study suggests that the pressure in extraction method and the extraction time affect the compositional constituents in BJIGT decoctions. The extraction time of 180 min could be chosen in both pressurized and non-pressurized method as optimal extraction condition.