• KSBB Journal
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• 제30권4호
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• pp.166-174
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• 2015

In this study, an efficient whole cell-based biotransformation for the production of liquiritigenin was developed using Laetiporus sulphureus CS0218 as biocatalyst and aqueous extracts of Glycyrrhiza uralensis as co-substrate, respectively. In order to determine the efficacy of this method, the optimal bioconversion conditions including mycelial growth, three important enzyme activities (${\beta}$-glucosidase, ${\alpha}$-rhamnosidase and ${\beta}$-xylosidase), and apparent viscosity of culture broth were monitored. After optimization, aqueous extracts of G. uralensis were added to the culture medium to directly produce algycone liquiritigenin. By applying this strategy, 67.5% of liquiritin was converted to liquiritigenin at pH 3.0 after 9 days of incubation and finally liquiritigenin was purified from the reaction mixture. And then, their biological activities including anti-oxidant and superoxide dismutase were observed. In fact, purified liquiritigenin was capable of bi-directional functions (i.e., either up-regulation or down-regulation of SIRT1 which is associated with aging). The results indicate that this strategy would be beneficial to produce biologically active liquiritigenin and could be used in pharmaceutical, cosmetic and food applications.

• 동의생리병리학회지
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• 제20권1호
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• pp.15-19
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• 2006

The anti-angiogenic activity of ethanol extract of Bojungbangam-tang, a new herbal prescription composed of nine crude drugs, was evaluated in human umbilical vein endothelial cells (HUVECs). HPLC profile revealed that five major compunds such as apioliquiritin, narirutin, hesperidin, liquiritin and glycyrrhizin. Ethanol extract of Bojungbangam-tang (EBJT) did not showed any significant cytotoxicity against HUVECs up to 200 ug/ml. EBJT significantly inhibited basic fibroblast growth factor (bFGF)-induced HUVECs proliferation to 69% at 200 ${\mu}g/ml$. Migration using window scraping method and tube formation in bFGF stimulated HUVECs were also significantly suppressed by EBJT in a dose-dependent manner. Taken together, these results suggest that Bojungbangam-tang can be a potent prescription for angiogenesis related disease.

• 동의생리병리학회지
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• 제29권2호
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• pp.168-173
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• 2015

Bangpungtongseong-san(BPTSS, Fangfengtongsheng-san in Chineses) is a traditional herbal formula comprising 18 medicinal herbs. In the present study, we performed the simultaneous analysis for four compounds of BPTSS and examined anti-allergic effects in human keratinocytes and mouse splenocytes. The column for separation of four compounds was used Luna C18 column and maintained at 40℃. The mobile phase for gradient elution consisted of two solvent systems. The analysis was carried out at a flow rate of 1.0 mL/min with PDA detection at 254 and 280 nm. To evaluate production and expression of Th2 chemokines, ELISA and RT-PCR were conducted in tumor necrosis factor (TNF)-α and interferon (IFN)-γ-stimulated HaCaT cells with or without BPTSS or silymarin, a positive control for skin inflammation. To measure Th2 cytokines, primary mouse splenocytes were treated with BPTSS and performed ELISA for interleukin (IL)-4, 5, 13. Calibration curves were acquired with r2>0.9999. The contents of geniposide, liquiritin, baicalin, and glycyrrhizin in BPTSS were 5.06 ㎎/g, 7.33 ㎎/g, 27.56 ㎎/g, and 7.81 ㎎/g, respectively. BPTSS reduced TARC and RANTES production and mRNA expression in TNF-α and IFN-γ-treated HaCaT cells. BPTSS inhibited IL-4, 5, and 13 production in mouse splenocytes. Our data will be a helpful information to upgrade quality control and anti-allergic effects of BPTSS.

• 대한한방부인과학회지
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• 제26권4호
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• pp.48-65
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• 2013

Objectives: We took breast milk samples and analyzed them using HPLC and LC/MS/MS, to evaluate the effects of taking Saenghwa-tang during breastfeeding on breast milk. Methods: The study participants were 20 lactating women who admitted in Korean medical postpartum care center. Breast milks were collected from paticipants who have been administrated Saenghwa-tang for more than 3 days. We used HPLC and LC/MS/MS for the determinations of amygdalin, liquiritins, 6-gingerol, decursin and decursinol angelate in Saenghwa-tang. Results: 1. Participants' $Mean{\pm}S.D$ (standard deviation) of age is $31.05{\pm}1.96$, and 15 participants had normal delivery and 5 participants had cesarean delivery. 12 participants were primipara and 8 participants were multipara. $Mean{\pm}S.D$ of lactating date is $9.4{\pm}0.94$. 2. Using HPLC, we learned LOQ level peak that matches the peak retention time of standard components of Saenghwa-tang was not detected from 20 breast milk samples. 3. Using LC/MS/MS, decursin of Angelicae Gigantis Radix was detected from HMSP 02, HMSP 04, HMSP 06, HMSP 11, and the each concentrations are 16, 2, 64, 11 ppb. Liquiritin of Glycyrrhizae Radix was not detected from HMSP 13~HMSP 18. Conclusions: Data obtained by this approach shows that this method is reliable and suitable for determining the safety of taking Saenghwa-tang during breastfeeding.

• 대한예방한의학회지
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• 제17권3호
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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.

• 생약학회지
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• 제44권2호
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• pp.125-130
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• 2013

Samchulgeonbi-tang (SC) is well-known traditional herbal medicine which is composed of fourteen medicine herbs. SC has been used for the treatment of the chronic gastritis, indigestion, gastric ulcers, gastroptosis and diarrhea disease. The variation in the amount of bioactive components of SC and its fermentation SC with ten Lactobacillus strains were investigated via high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). Simultaneous qualitative and quantitative analysis of seven bioactive compounds; paeoniflorin, liquiritin, hesperidin, liquiritigenin, kaempferol, atractylenolide III, magnolol were achieved by comparing their retention times ($t_R$) and UV spectra with those of the standard compounds. In the results, the amount of paeoniflorin and hesperidin were 7.967 mg/g, 7.251 mg/g that were the main compounds in SC. The amounts of liquiritigenin was increased by all ten Lactobacillus strains, except strain 128. Especially, the liquiritigenin amount was highest in SC fermented with strain 145 (0.201 mg/g), which was increased by 158.39% compared to SC (0.081 mg/g). In the fermented SC using strains 344, almost components were increased than non-fermented SC, except paeoniflorin and kaempferol. Thus, these results considered that the strains 145 and 344 are most excellent fermentation strains among the 10 species of fermentation strains.

• 한국약용작물학회지
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• 제19권6호
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• pp.446-455
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• 2011

Galgeun-tang (GGT) is a traditional medicinal formula that is widely prescribed to treat cold, asthma, and hives in Korea. Fermented herbal medicines can be made more effective than normal herbal medicines by increasing the absorption and bioavailability of the active compounds. In this study, we fermented Galgeun-tang to produce bioconversion constituents using Lactobacillus plantarum (GGT144), and found that four peaks were decreased, three peaks were increased and two new peaks appeared in the HPLC-DAD chromatogram. After HPLC-DAD-guided fractionation of the newly-appearing compounds (1 and 5) and the increased (6, 7, and 9) compounds, the structure of the compounds was determined using NMR and MS. Using this approach the compounds were identified to be pyrogallol (1), daidzein (5), liquiritigenin (6), cinnamyl alcohol (7), and formononetin (9), respectively. In addition, the decreased compounds were identified to be daidzin (2), liquiritin (3), ononin (4), and cinnam aldehyde (8) using HPLC-DAD analysis with standard compounds. The high performance liquid chromatography method was used to quantify the nine constituents in GGT and GGT144. All calibration curves of the standard compounds displayed excellent linearity with a $R^2$ > 0.9968.

• 동의생리병리학회지
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• 제28권3호
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• pp.322-329
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• 2014

The purpose of this study was to investigate the quality of two different commercial Bangpungtongseong-san (BTS) extract granules (BTS-2 and BTS-3) by comparing with BTS decoction (BTS-1). The contents of characterizing components and biological activities of two different commercial BTS extract granules were compared with those of the BTS decoction. The contents of characterizing components were analyzed with HPLC. The antioxidative effects were determined by measuring 2,2-diphenyl-1-picrylhygrazyl (DPPH) radical scavenging and superoxide dismutase (SOD)-like activity. Also, we compared the effects on lipid accumulation and reactive oxygen species (ROS) production during differentiation of 3T3-L1 preadipocytes. The contents of five components except liquiritin and sennoside A were higher in BTS-1. The DPPH radical scavenging and SOD-like activity were higher in BTS-1. BTS-1 significantly inhibited lipid accumulation during differentiation of 3T3-L1 preadipocytes and showed stronger effects than BTS-2, BTS-3. In addition BTS-1 showed stronger inhibition effects on ROS production during differentiation of 3T3-L1 preadipocytes than BTS-2, BTS-3. These results indicate that BTS decoction has strong biological activities than commercial BTS extract granules. It is also consistent with the contents of characterizing components.

• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.178-186
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• 2020

Licorice (Glycyrrhiza uralensis) contains several compounds that have been reported to alleviate menopausal symptoms via interacting with estrogen receptors (ERs). The compounds exist mainly in the form of glycosides, which exhibit low bioavailability and function. To bioconvert liquiritin and isoliquiritin, the major estrogenic compounds, to the corresponding deglycosylated liquiritigenin and isoliquiritigenin, respectively, licorice was fermented with Monascus, which has been demonstrated to deglycosylate other substances. The contents of liquiritigenin and isoliquiritigenin in Monascus-fermented licorice increased by 10.46-fold (from 38.03 μM to 379.75 μM) and 12.50-fold (from 5.53 μM to 69.14 μM), respectively, compared with their contents in non-fermented licorice. Monascus-fermented licorice exhibited 82.5% of the ERβ binding activity of that observed in the positive control (17 β-estradiol), whereas the non-fermented licorice exhibited 54.1% of the binding activity in an in vivo ER binding assay. The increase in the ERβ binding activity was associated with increases in liquiritigenin and isoliquiritigenin contents. Liquiritigenin acts as a selective ligand for ERβ, which alleviates menopausal symptoms with fewer side effects, such as heart disease and hypertension, compared with a ligand for ERα. In addition, Monascus-fermented licorice contained 731 mg/kg of monacolin K, one of the metabolites produced by Monascus that reduces serum cholesterol. Therefore, Monascus-fermented licorice is a promising material for the prevention and treatment of menopausal syndrome with fewer side effects.

• 대한한의학방제학회지
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• 제21권1호
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• pp.71-79
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• 2013

Objectives : Yijin-tang has been used in the treatment of gastrointestinal diseases such as irritable bowel syndrome, gastritis, and gastric ulcer. In this study, a high-performance liquid chromatography (HPLC) method was established for simultaneous analysis of six compounds, liquiritin, glycyrrhizin, hesperidin, 6-gingerol, homogentisic acid, and 3,4-dihydroxybenzaldehyde in Yijin-tang, a traditional Korean herbal medicinal preparation. Methods : A Gemini C18 column was used for the separation of six constituents at $40^{\circ}C$. The mobile phase using gradient elution consist of two solvent systems, 1.0% acetic acid in water (A) and 1.0% acetic acid in acetonitrile (B). The flow-rate was 1.0 mL/min and injection volume was $10{\mu}g$. The detector was a photodiode array (PDA) detector set at 254 nm and 280 nm. Results : The calibration curves of six compounds showed good linearity in various concentration ranges ($R^2{\geq}0.9997$). The limits of detection (LOD) and limits of quantification (LOQ) were 0.028-$0.192{\mu}g/mL$ and 0.093-$0.540{\mu}g/mL$, respectively. The RSD (%) of the intra and inter day validations were 0.03-0.84% and 0.05 -1.00%, respectively. Recovery was 96.14-01.90% and RSD (%) was less than 1.5%. Conclusions : The established simultaneous analysis methods will help management to improve the quality of Yijin-tang.