• 생약학회지
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• 제41권3호
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• pp.161-165
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• 2010

A study was performed to assess the quality equivalence of commercially available Hwang-Ryun-Hae-Dok-Tang granules from various vendors in terms of the contents of characterizing compounds. 'Hwang-Ryun-Hae-Dok-Tang'(or 'Huanglian Jiedu Tang') is the complex medicinal herbal product which is composed of four different herbal drugs; Scutellariae Radix, Coptidis Rhizoma, Phellodendri Cortex and Gardeniae Fructus. Using a high performance liquid chromatography and direct analysis in real time mass spectrometer (DART-MS), five characterizing compounds such as baicalin, baicalein, berberine, palmatine, and geniposide were analyzed to compare the variance of the contents between the products from four different vendors. Although five characterizing compounds were all successfully detected from all the products, the contents of each compound were shown to have significant difference among vendors. For real time monitoring of characterizing compounds in medicinal herbal products, DART-MS can be successfully adopted as a powerful analytical tool.

• Toxicological Research
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• 제21권3호
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• pp.255-261
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• 2005

Gami-Samhwang-San, a herbal prescription for obesity treatment, is composed of seven crude herbs such as Rehmanniae Radix Preparata, Ephedrae Herba, Scutellariae Radix, Acori Gramineri Rhizoma, Polygalae Radix, Typhae Pollen, Armeniacae Semen, Menthae Herba. In this study, marker substances in n-butanol fraction (SH-21-B) from Gami-Samhwang-San were analyzed by high performance liquid chromatography (HPLC) and acute toxicity of standardized SH-21-B was evaluated by good laboratory practices (GLP) guideline of Korea Food and Drug Administration. Therefore we confirmed that there were baicalin of 15.92%, amygdalin of 6.57% and ephedrine of 2.49% in SH-21-B. SH-21-B was administered in rats at dose of 0 mg/kg, 2,000 mg/kg, and 5,000mg/kg. Clinical signs of both sexes of rats were observed daily for 14 days after single oral administration. Two female rats one administered at 2,000 mg/kg and the other administered at 5,000 mg/kg, died, but no dead animal was observed among male rats. Therefore $LD_{50}$ in the female rat is observed to be 8,710 mg/kg, and MLD (Minimun Lethal Dose) of the male rat is observed to be more than 5,000 mg/kg.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.72-72
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• 2020

Various mixtures were prepared depending on the mixing ratio of Scutellaria baicalensis hot water extract (SB-HW) and Chrysanthemum morifolium ethanol extract (CM-E) and their anti-inflammatory activity were compared. Among them, SB-HW (80 ㎍/mL)/CM-E (120 ㎍/mL) or SB-HW (40 ㎍/mL)/CM-E (160 ㎍/mL) significantly inhibited LPS-stimulated NO and IL-6 levels in RAW 264.7 cells. The SB-HW (80 ㎍/mL)/CM-E (120 ㎍/mL) mixture, which was determined as active mixture, significantly reduced MUC5AC secretion in PMA and LPS-induced NCI-H292 cells. The active mixture also reduced the production of PGE2 and IL-8 in PMA-induced A549 cells. LC-MS/MS analysis showed that the active mixture was composed of high contents of flavone glycosides, such as baicalin and cynaroside. Western blot analysis indicated that the active mixture suppressed phosphorylation of ERK, JNK, and p38, associating with the inhibition of MAPK signaling. Taken together, our results suggest that the active mixture could be applied as a new anti-inflammatory herbal medicine

• 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.

• 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.

• 한국약용작물학회지
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• 제14권2호
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• pp.117-123
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• 2006

The purpose is to study the identification method of Huangjinju powder for injection and the medicinal materials by the fingerprint off-ray Diffraction Fourier (XRDF). We used the same method on both the studying of Huangjinju and the medicinal materials. Then we selected a few components alignment to compare. We analyzed the data by setting up the deviation $d({\AA})$ as ${\pm}0.05$ to calculate the rate of special mark on the sample (Px) and on the patent (P). The special XRDF of Huangjinju$[d({\AA})/(I/I_0)]$ have 5 peaks that have not expressed in medicinal materials. Therefore Px is 22.73%. Flos Trollii Chinensis has 3 special marks and Px is 17.65%. Flos Chrysanthemi Indici has 1 special mark and Px is 3.57%. Its coincided interplanar spacing with the patent is $2.907{\AA}$. Flos Lonicerae Japonicae has 6 special marks and Px is 23.08%. Its special mark in the patent are 4.95/14 and 4.50/15, respectively. The P is 9.09%. Its coincided interplanar spacing with the patent is $2.910{\AA}\;and\;3.05{\AA}$, respectively. The number of special XRDF mark peaks of baicalin is 9 and Px is 18.37%. Its coincided interplanar spacing with the patent is $2.910{\AA}$. It has visible mark and specificity adopting XRDF fingerprint to identify Huangjinju and medicinal materials. Establishing the quality standard is a synthetic index that depends both on special marks in the medicinal materials of the patent and on the coincidence peak data.

• Natural Product Sciences
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• 제22권4호
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• pp.238-245
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• 2016

Gumiganghwal-tang has been used for the treatment of common cold for a long-time. We developed an accurate and sensitive high performance liquid chromatography-diode array detection (HPLC-DAD) and electrospray ionization mass spectrometry method for the simultaneous determination of ferulic acid, baicalin, bergapten, methyl eugenol, glycyrrhizin, oxypeucedanin, wogonin, nodakenin, atractylenolide III, imperatorin, and atractylenolide I in Gumiganghwal-tang samples. The analytes were separated on a Shiseido C18 column ($5{\mu}m$, $4.6mm\;I.D.{\times}250mm$) with gradient elution with acetonitrile and 0.1% trifluoroacetic acid. Eleven compounds were quantitatively determined by HPLC-DAD and identified by LC-MS data. We also validated this method. The calibration curves of all the compounds showed good linear regression. The limits of detection and the limits of quantification ranged from 0.04 to 0.63 and from 0.12 to $1.92{\mu}g/mL$, respectively. The relative standard deviation values of intra- and inter-days of this method represented less than 2.9%. The recoveries were found to be in the range of 90.06 - 107.66%. The developed method has been successfully applied to the analysis of Gumiganghwaltang samples. The established HPLC method could be used to quality control of Gumiganghwal-tang.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.792-799
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• 2005

Two novel $\beta$-glucuronidases, which metabolize glycyrrhizin (GL) to 18$\beta$-glycyrrhetinic acid-3-O-$\beta$-D-glucuronide (GAMG), were purified from Streptococcus LJ-22 isolated from human intestinal microflora. $\beta$-Glucuronidases I and II were purified to apparent homogeneity, using a combination of ammonium sulfate fractionation, butyl toyopearl, Q-Sepharose, hydroxyapatite Ultrogel, and GL-attached Sepharose column chromatographies, with the final specific activities of 137 and 190 nmole/min/mg, respectively. The molecular sizes of both $\beta$-glucuronidases were found to be 140 kDa by gel filtration, and they consisted of two identical subunits (M.W. 67 kDa by SDS-PAGE). $\beta$-Glucuronidases I and II showed optimal activity at pH 7.0 and pH 6.5, respectively. Both purified enzymes were potently inhibited by $Cu^{2+}$ and PCMS, and had maximum activity on glycyrrhizin, but did not hydrolyze p-nitrophenyl-$\beta$-glucuronides, baicalin, or GAMG These findings suggest that the biochemical properties and substrate specificities of these enzymes are different from those of the previously purified $\beta$-glucuronidases. This is the first reported purification of sugar (not aglycone)-recognizing $\beta$-glucuronidases from intestinal bacteria.

• Natural Product Sciences
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• 제20권4호
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• pp.251-257
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• 2014

Chungsimyeonja-tang (CSYJT) is an herbal decoction that consists with 12 herbal medicines. CSYJT has been shown to have anti-stress, anti-allergic and anti-oxidant effects. The aim of this study was to determine flavonoid contents in CSYJT and evaluate its protective effect against cisplatin induced nephrotoxicity using both in vitro (porcine renal epithelial cell; PK15 cell) and in vivo (Sprague Dawley rat) experiments. In the present study, thee mean contents of baicalin, wogonoside and baicalein in CSYJT were 14.65, 5.27 and 0.02 mg/g, respectively. The CSYJT extract treatment attenuated the following alteration in porcine renal epithelial (PK15) cell: the increase in reactive oxygen species (ROS), the glutathione depletion and the increase in p53 expression induced by cisplatin treatment. In the in vivo study, rats were orally treated with CSYJT extract once a day for 28 days. Five days before the last treatment, cisplatin (5 mg/kg) was intraperitoneally injected to induce acute renal failure. Increased blood urea nitrogen (BUN) and creatinine (CRE) levels after cisplatin treatment were ameliorated by pretreatment of CSYJT extract. In addition, lipid peroxidation was decreased and antioxidant enzyme (glutathione) was recovered in CSYJT pretreated kidney tissue. In histopathological examination, CSYJT pretreated group showed ameliorated pathological alteration after cisplatin injection with decreased apoptosis. Taken together, pretreatment of CSYJT could ameliorate cisplatin-induced nephrotoxicity.

• 동의생리병리학회지
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• 제20권2호
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• pp.455-459
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• 2006

The inhibitors of prostaglandin $E_2\;(PGE_2)$ production and cyclooxygenase-2 (COX-2) activity have been considered as potential anti-inflammatory agents. In this study, we evaluated 9 compounds isolated from 5 Korean phytomedicinal plants (Spirea prunifolia, Paeonia suffruticosa, Salvia miltiorrhiza, Scutellaria baicalensis, and Artemisia capillaris) for the inhibition of $PGE_2$production and COX-2 expession in lipopolysaccharide (LPS)-stimulated human macrophages U937 cells. As a result, several compound such as prunioside A, penta-O-galloyl-beta-D-glucose, tanshinone IIA, baicalin, baicalein, wogonin, scopolatin, scoparone and decursinol showed potent inhibition of $PGE_2$production (50-70% inhibition at the test concentration of $10\;{\mu}M$). In addition, these compounds were also considered as potential inhibitors of COX-2 activity (45-73% inhibition at the test concentration of $10\;{\mu}M$). These active compound mediating COX-2 inhibitory activities are warranted for further elucidation of active principles for development of anti-inflammatory agents and these properties may contribute to the anti-atopic dermatitis activity.