• Korean Journal of Pharmacognosy
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• v.45 no.3
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• pp.249-255
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• 2014

This study compared Bojungikgi-tang (BIT) and Yukmijihwang-tang (YJT) decoctions produced using the extractors made of various materials, such as glass, earthenware, and stainless steel, for 60, 90, and 120 min. The extraction yield was measured and the amounts of marker compounds in decoctions were determined by quantitative analysis using high performance liquid chromatography. The extraction yields of BIT decoctions produced by three extractors were not significantly different, whereas the extraction yield of YJT produced using glass extractor was significantly higher than that of YJT produced using earthenware extractor after 90 min. The amounts of most marker compounds were highest in BIT decoctions produced using glass extractor. However, variations of the amounts of marker compounds were observed in YJT decoctions and the volatile compound was not easily vaporized in YJT decoction produced using earthenware extractor. This study suggests that the extractor made of various materials may affect the extraction efficiency and the contents of marker compounds, especially in the decoction produced using earthenware extractor, although the results were dependent on the characteristics of herbal formula.

• Journal of Society of Preventive Korean Medicine
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• v.17 no.3
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• pp.165-176
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• 2013

Objective : The purpose of this study was to establish the simultaneous analysis for six compounds in Gumiganghwal-tang (GMGHT, Jiuweiqianghuo-tang) and to investigate the anti-atherosclerotic effects of GMGHT in vitro. Methods : The column for separation of six compounds was used Luna $C_{18}$ column and maintained at $40^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems, 1.0% acetic acid in water and 1.0% acetic acid in acetonitrile. The analysis was carried out at a flow rate of 1.0 mL/min with pothodiode array (PDA) detection at 254, 280, and 320 nm. The injection volume was 10 ${\mu}L$. The antioxidant activities of GMGHT were evaluated by measuring free radical scavenging activities on 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS), relative electrophoretic mobility (REM), and fragmentation of apolipoprotein B (ApoB)-100. Results : Calibration curves were acquired with $r^2{\geq}0.9998$. The contents of liquiritin, ferulic acid, baicalin, baicalein, glycyrrhizin, and wogonin in GMGHT were 1.784, 1.693, 37.899, 0.258, 1.869, and 0.034 mg/g, respectively. The GMGHT showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were 72.51 ${\mu}g/mL$ and 128.49 ${\mu}g/mL$. Furthermore, GMGHT reduced the oxidation properties of LDL induced by $CuSO_4$. Conclusion : HPLC-PDA is considered as an available and convenient method for quality control and standardization of GMGH and GMGHT has potentials on anti-atherosclerosis by anti-oxidative effect and suppressive effect on LDL oxidation.

• The Journal of Korean Medicine
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• v.34 no.4
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• pp.12-20
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• 2013

Objectives: This study compared Gumiganghwal-tang (GGT) decoctions produced using different pressure levels for various extraction times to determine the optimal extraction conditions through hydrogen ion concentration (pH), total soluble solids content (TSSC), extraction yield, and content of chemical compounds. Methods: Decoctions were prepared by the pressure levels of 0 or $1kgf/cm^2$ for 30-180 min. The pH and TSSC were measured, the extraction yield was calculated, and the amounts of the chemical compounds were determined using high performance liquid chromatography. Results: The higher pressure and longer extraction time decreased the pH value, while those conditions increased TSSC and extraction yield: the decoction produced in 180 min by pressurized method showed the minimum value of pH, but maximum values of TSSC and extraction yield. The chemical compounds showed higher amounts in decoctions produced by non-pressurized methods than pressurized methods and their amounts were decreased over the peak extraction time in both pressurized and non-pressurized methods. The results of regression analysis confirmed the correlative influences of the pressure and extraction time on pH, TSSC, and extraction yield. Conclusions: This study suggests that pressure and extraction time influence the compositional constituents in GGT decoctions, and the non-pressurized method for 120 min should be chosen as the optimal extraction condition for the preparation of GGT decoction.

• Journal of Haehwa Medicine
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• v.23 no.1
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• pp.93-104
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• 2014

Glycyrrhizae Radix et Rhizoma has been extensively used by human beings as a medicinal herb as well as natural sweetener. In this study, we performed quantification analysis of three major constituents including liquiritin, glycyrrhizin, and glycyrrhetinic acid in the 70% ethanol extracts of non-processed Glycyrrhizae Radix et Rhizoma and processed Glycyrrhizae Radix et Rhizoma using a high-performance liquid chromatography coupled with photodiode array detector. The analytical column for separation of the 3 constituents used a Gemini C18 column kept at $40^{\circ}C$ by the gradient elution of two mobile phase. The amounts of liquiritin, glycyrrhizin, and glycyrrhetinic acid in non-processed Glycyrrhizae Radix et Rhizoma were 2.57%, 3.52%, and not detected. After processing by roasting, the best roasting temperature and time of iquiritin, glycyrrhizin, and glycyrrhetinic acid were $160^{\circ}C$-15 min (2.46%), $160^{\circ}C$-15 min (3.67%), and $240^{\circ}C$-15 min (0.76%), respectively.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.392-402
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• 2017

Background: Previously, we reported that Korean Red Ginseng inhibited liver fibrosis in mice and reduced the expressions of fibrogenic genes in hepatic stellate cells (HSCs). The present study was undertaken to identify the major ginsenoside responsible for reducing the numbers of HSCs and the underlying mechanism involved. Methods: Using LX-2 cells (a human immortalized HSC line) and primary activated HSCs, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assays were conducted to examine the cytotoxic effects of ginsenosides. $H_2O_2$ productions, glutathione contents, lactate dehydrogenase activities, mitochondrial membrane permeabilities, apoptotic cell subpopulations, caspase-3/-7 activities, transferase dUTP nick end labeling (TUNEL) staining, and immunoblot analysis were performed to elucidate the molecular mechanism responsible for ginsenoside-mediated cytotoxicity. Involvement of the AMP-activated protein kinase (AMPK)-related signaling pathway was examined using a chemical inhibitor and small interfering RNA (siRNA) transfection. Results and conclusion: Of the 11 ginsenosides tested, 20S-protopanaxadiol (PPD) showed the most potent cytotoxic activity in both LX-2 cells and primary activated HSCs. Oxidative stress-mediated apoptosis induced by 20S-PPD was blocked by N-acetyl-$\text\tiny L$-cysteine pretreatment. In addition, 20S-PPD concentration-dependently increased the phosphorylation of AMPK, and compound C prevented 20S-PPD-induced cytotoxicity and mitochondrial dysfunction. Moreover, 20S-PPD increased the phosphorylation of liver kinase B1 (LKB1), an upstream kinase of AMPK. Likewise, transfection of LX-2 cells with LKB1 siRNA reduced the cytotoxic effect of 20S-PPD. Thus, 20S-PPD appears to induce HSC apoptosis by activating LKB1-AMPK and to be a therapeutic candidate for the prevention or treatment of liver fibrosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.32 no.6
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• pp.384-395
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• 2018

The aim of this study was to investigate whether a novel formulation of an herbal extracts has an inhibitory effect on obesity. To determine its anti-obesity effects, we performed anti-obesity-related experiments in vitro and in vivo. Thus, our present study was carried out to evaluate the anti-obesity effect of herbal extracts using a high fat diet (HFD)-induced obese mouse model and 3T3-L1 adipose cells. The effects of each herbal extracts on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. Results showed that treatment with each herbal extracts at $10{\sim}100{\mu}g/ml$ had no effect on cell morphology and viability. Without evidence of toxicity, herbal extracts treatment decreased lipid accumulation compared with the untreated adipocytes controls as shown by the lower absorbance of Oil Red O stain. Futhermore, compared with control-differentiated mature adipocytes, each herbal extracts significantly inhibited lipid accumulation in mature 3T3-L1 adipocytes. In the HFD-fed obese mice, body weight, liver weight and white adipose tissue weights were significantly reduced by mixture of herbal extracts administration in mouse skin. Futhermore, we found that mixture of herbal extracts administration suppressed serum triglyceride (TG), and total cholesterol (TCHO) in HFD-induced obese mouse model. The mixture of herbal extracts of permeability was estimated by measuring the transepithelial electrical resistance (TEER) value in pig skin. The optimized formulations of herbal extracts (Test 3 formulation) showed skin permeation. However, test 1 formulation containing essential oil as enhancer showed maximum skin permeation. After confirming the enhanced skin permeability, in vivo studies were performed to assess whether skin irritation potential on the basis of a primary irritation index (PII) in rabbit skin. Reactions were scored for erythema/edema reactions at 24 h, 48 h and 72 h post-application. It was concluded that the test 1 formulation was not irritation (PII = 0). The present study suggests that the test 1 formulation might be of therapeutic interest with respect to the treatment of obesity.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.1-6
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• 2014

Objectives : This study was aimed to compare Gyejibokryeong-hwan (GBH) decoctions produced using different pressure levels for various extraction times to find the optimal extraction conditions through extraction yield, total soluble solids content (TSSC), hydrogen ion concentration (pH), and the contents of chemical compounds. Methods : Decoctions of GBH were prepared under the pressure levels of 0 or $1kgf/cm^2$ for 30-180 min using water as extraction solvent. The extraction yield, TSSC, and pH were measured, and the amounts of the chemical compounds were determined using high performance liquid chromatography-photodiode array detector. Results : The higher pressure and longer extraction time increased the values of TSSC and extraction yield, while decreased the pH value. The decoctions produced in 180 min by pressurized method and produced in 150 min by non-pressurized method showed maximum values of extraction yield and TSSC with minimum value of pH. The amounts of chemical compounds showed variations in pressurized and non-pressurized decoction during overall extraction times. The influences of pressure and extraction time on extraction yield, TSSC, pH, and the contents of chemical compounds were confirmed by regression analysis, which showed that all extraction values were significantly affected by at least one of two extraction factors, pressure and extraction time. Conclusions : This study suggests that the pressure and extraction time can significantly affect the extraction efficiency of components from GBH decoctions. However, optimal extraction conditions could not be chosen due to the variation of the amounts of chemical compounds.

• The Korea Journal of Herbology
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• v.28 no.4
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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.

• Korean Journal of Pharmacognosy
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• v.47 no.4
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• pp.360-365
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• 2016

The aim of this study was to compare the amount of spinosin in the 70% ethanol extracts of non-processed Zizyphi Semem (ZS) and processed ZS by roasting using a high-performance liquid chromatography equipped with photodiode array detector. Separation of the spinosin was used $SunFire^{TM}$ $C_{18}$ analytical column ($5{\mu}m$, $4.6{\times}150mm$) using two mobile phase consisting of distilled water and acetonitrile, both with 1.0% (v/v) acetic acid. The flow rate was 1.0 mL/min and injection volume was $10{\mu}L$. Calibration curve of the spinosin was y = 22339.45x+483.99 in tested concentration range ($1.28-20.00{\mu}g/mL$) and correlation coefficient was 1.0000. In non-processed ZS sample, the amount of the spinosin was 0.94 m/g, while, the amount of the marker compound in processed ZS samples were 0.66-1.10 mg/g.

• Korean Journal of Pharmacognosy
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• v.48 no.3
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• pp.226-231
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• 2017

Zingiberis Rhizoma (ZR, Zingiberaceae) is one of the foods for a long time and contains various biological activities including anti-cancer, anti-inflammatory, anti-oxidant, and anti-platelet activities. The present study was to compare the amount of 6-gingerol in the 70% ethanol extracts of non-processed ZR and processed ZR by roasting using a high-performance liquid chromatography equipped with photodiode array detector. The 6-gingerol was separated on a Gemini $C_{18}$ analytical column ($5{\mu}m$, $4.6{\times}250mm$) using two mobile phase consisting of distilled water and acetonitrile. The flow rate was 1.0 mL/min and injection volume was $10{\mu}L$. In non-processed ZR sample, the amount of the 6-gingerol was 0.43% and the amount of the marker compound in processed ZR samples by roasting were 0.16-0.55%.