• Natural Product Sciences
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• v.8 no.2
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• pp.35-43
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• 2002

Glycosides of herbal medicines, such as glycyrrhizin, ginsenosides, kalopanaxsaponins, rutin and ponicirin, were studied regarding their metabolic fates and pharmacological actions in relation to intestinal bacteria using germ-free, gnotobiotic and conventional animals. When glycyrrhizin (GL) was orally administered, $18{\beta}-glycyrrhetinic\;acid\;(GA)$, not GL, was detected in plasma and intestinal contents of gnotobiotic and conventional rats. However, GA could not be detected in germ-free rats. When GL was incubated with human intestinal bacteria, it was directly metabolized to GA (>95%) or via $18{\beta}-glycyrrhetinic\;acid-3-{\beta}-D-glucuronide$(>5%). Orally administered GL was effective in gnotobiotic and conventional rats for liver injury induced by carbon tetrachloride, but was not effective in germ-free rats. When ginseng saponins were orally administered to human beings, compound K in the plasma was detected, but the other protopanxadiol saponins were not detected. The compound K was active for tumor metastasis and allergy. When kalopanaxsaponins were incubated with human intestinal microflora, they were metabolized to kalopanaxsaponin A, kalopanaxsaponin I and hederagenin. These metabolites were active for rheumatoid arthritis and diabetic mellitus while the other kalopanxsaponins were not. When flavonoid glycosides were orally administered to animals, aglycones and/or phenolic acids were detected in the urine. The metabolic pathways proceeded by intestinal bacteria rather than by liver or blood enzymes. These metabolites, aglycones and phenolic acids, showed antitumor, antiinflammatory and antiplatelet aggregation activities. These findings suggest that glycosides of herbal medicines are prodrugs.

• Korean Chemical Engineering Research
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• v.53 no.3
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• pp.321-326
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• 2015

Plackett-Burman (PB) and Box-Behnken (BB) experimental designs were used to optimize fermentation variables for the biotransformation of glycyrrhizin (GL) to monoglucuronyl-glycyrrhetinic acid (MGGA). The PB design was first used to screen the important factors among the medium variables. The steepest ascent method was used to approach the optimum range for each of these factors. The BB design was finally used to analyze the response surfaces of the screened factors for further optimization. The optimized conditions for this system were 0.7 g/L $MgSO_4{\cdot}7H_2O$, 1.19 g/L GL, and cultivation for six days. The biotransformation of GL to MGGA could reach up to 35.72%, which is a good result for this kind of transformation.

• Korean Journal of Pharmacognosy
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• v.42 no.3
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• pp.240-245
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• 2011

A high-performance liquid chromatography (HPLC) method was developed for quantitative analysis of liquiritin and glycyrrhizin in Sagunja-tang (SGT, Sijunzi-tang in Chinese), a traditional Korean medicine. HPLC analysis was performed using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 254 nm and 280 nm for quantification of the two components in SGT. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. Calibration curves were acquired with $r^2$ values > 0.9998, and the relative standard deviations (RSDs, %) for intra- and inter-day precision were not exceed 4.0%. The recovery of each component was in the range of 91.85 - 108.62%, with a RSD less than 4.0%. The contents of the two components in SGT were 7.94 - 13.83 mg/g.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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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.

• YAKHAK HOEJI
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• v.57 no.3
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• pp.187-193
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• 2013

Bojungikgi-tang has been widely used for enhancement of physical fitness in Korea. The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of four marker compounds, liquiritin, nodakenin, hesperidin, and glycyrrhizin in Bojungikgi-tang (Buzhongyiqi-tang in Chinese, Hochuekkito in Japanese), a traditional Korean herbal prescription. The column for optimizing HPLC separation was used a Gemini $C_{18}$ column at column oven temperature of $40^{\circ}C$ with 1.0% (v/v) aqueous acetic acid (A) and 1.0% (v/v) acetic acid in acetonitirle (B) by gradient flow. The flow rate was 1.0 ml/min and the detector was a photodiode array (PDA) set at 254 nm, 280 nm, and 335 nm. Calibration curves of four components were acquired with $r^2$ values ${\geq}0.9999$. The recoveries were found to range 92.11~105.68% with relative standard deviations (RSDs, %) value less than 2.50%. The RSD values of intraand inter-day precision were 0.07~2.50% and 0.16~1.99%, respectively. The contents of liquiritin, nodakenin, hesperidin and glycyrrhizin in Bojungikgi-tang were 3.85~3.92 mg/g, 2.27~2.32 mg/g, 4.14~4.19 mg/g, and 3.39~3.45 mg/g, respectively. The established simultaneous analysis method will be effective for quality control of Bojungikgi-tang.

• YAKHAK HOEJI
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• v.55 no.6
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• pp.451-455
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• 2011

So Cheong Ryong Tang (SCRT) has been used as a traditional herbal medicine for the treatment of the bronchial asthma. In this study, the effect of steaming process on changes in 11 major compounds (homogentisic acid, ephedrine HCl, paeoniflorin, cinnamic acid, cinnam aldehyde, glycyrrhizin, 6-gingerol, schizandrin, methyleugenol, gomisin A and gomisin N) content and neuroprotective activity of SCRT were evaluated. Major compound content was slightly different with steaming produce. The contents of paeoniflorin, cinnamic acid, cinnam aldehyde, 6-gingerol and methyleugenol were decreased and homogentisic acid, ephedrine HCl, glycyrrhizin, schizandrin and gomisin A were increased by steaming process. The neuroprotective activity of steaming SCRT was determined in mouse hippocampal HT22 cells by MTT assay. As a result, neuroprotective activity of a steaming SCRT was higher than that of a non steaming SCRT. This study demonstrated that neuroprotective activity of SCRT can be improved through steaming process.

• Natural Product Sciences
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• v.14 no.3
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• pp.147-151
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• 2008

A high performance liquid chromatographic (HPLC) method for the simultaneous determination of marker constituents, baicalin and glycyrrhizin was established for the quality control of traditional herbal medicinal preparation, Eul-Ja-Tang (EJT). Separation and quantification were successfully achieved with a Waters XTerra RP18 column ($5{\mu}m$, 4.6 mm I.D. ${\times}$ 150 mm) by gradient elution of a mixture of acetonitrile and water containing 0.03% phosphoric acid (pH 2.03) at a flow rate of 1.0 mL/min. The diode-array UV/VIS detector (DAD) was used for the detection and the wavelength for quantification was set at 250 nm. The presence of baicalin and glycyrrhizin in this decoction was ascertained by retention time, spiking with each authentic standard and UV spectrum. Both baicalin and glycyrrhizin showed good linearity ($r^2$ > 0.999) in a relatively wide concentration ranges. The R.S.D. for intra-day and inter-day precision was less than 5% and the limits of detection (LOD) were about 30 ng. The mean recovery of each compound was 99.5 - 101.2% with R.S.D. values less than 4.0%. This method was successfully applied to the determination of contents of baicalin and glycyrrhizin in three commercial products of EJT, which resulted in the difference in the contents of these compounds. These results suggest that the developed HPLC method is simple, effective and could be readily utilized as a quality control method for commercial EJT products.

• Natural Product Sciences
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• v.16 no.1
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• pp.26-31
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• 2010

High performance liquid chromatographic method with diode-array detection (HPLC-DAD) has been performed for the simultaneous determination of four marker constituents, paeoniflorin, trans-cinnamic acid, schisandrin and glycyrrhizin in traditional herbal medicinal preparation, So-Cheong-Ryong-Tang (SCRT). The presence of paeoniflorin, trans-cinnamic acid, schisandrin and glycyrrhizin in this decoction was ascertained by retention time, spiking with each authentic standard, UV spectrum and ESI mass spectrum. All four compounds showed good linearity ($r^2$ > 0.998) in a relatively wide concentration ranges. The RSD for intra-day and inter-day precision was less than 3% and the limits of detection (LOD) were less than 30 ng. The mean recovery of each compound was 94.1-113.0% with RSD values less than 3.0%. These results suggest that the developed HPLC method is simple, effective and could be readily utilized as a quality control method for commercial SCRT products.

• YAKHAK HOEJI
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• v.35 no.3
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• pp.174-181
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• 1991

These experiments were conducted to investigate the effects of glycyrrhizin(GL) and glycyrrhetinic acid(GA) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [sup 3/H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}$Ca$^{2+}$ to P388D$_{1}$, cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GL(10$^{-3}$M) and GA(10$^{-4}$M). Histamine production in mouse spleen cell culture was significantly increased by the addition of 0.25 $\mu\textrm{g}$/ml of Con A, after 48 hour incubation. Con A dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 .mu.g/ml of Con A. The effects of GL on histamine contents and T-lymphocyte proliferation were significantly decreased at high dose (10$^{-5}$M), while IL-1 activity was remarkably suppressed by 10$^{-8}$~10$^{-4}$M of GL. $Ca^{2+}$ uptake was not changed, but antibody production was increased by GL(10 mg/kg). GA inhibited histamine contents at 10$^{-9}$~10$^{-7}$ and depressed Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-7}$~10$^{-5}$M of GA, but increased suboptimal dose (Con A 0.1 $\mu\textrm{g}$/ml) at 10$^{-9}$~10$^{-7}$M of GA. IL-1 activity was suppressed by 10$^{-8}$~10$^{-4}$M of GA and $Ca^{2+}$ uptake was enhanced by 10$^{-9}$~10$^{-6}$ of GA, but antibody production was not changed by GA. From the above results, it is suggested that GL and GA have immuno-regulatory action. GL decreased cell-mediated immune response, and increased humoral immune response at high dose. On the other hand, low dose of GA enhanced cell-mediated immune response, while high doses of GA decreased humoral immune reaction.

• Archives of Pharmacal Research
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• v.23 no.2
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• pp.172-173
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• 2000

The relationship between the metabolites of glycyrrhizin (18$\beta$-glycyrrhetinic acid-3-O--D-glu-curonopyranosyl-($1{\rightarrow}2$)-$\beta$-D-glucuronide, CL) and their biological activities was investigated. By human intestinal microflora, CL was metabolized to 18$\beta$-glycyrrhetinic acid (GA) as a main product and to 18$\beta$-glycyrrhetinic acid-3-O-$\beta$-D-glucuronide (GAMG) as a minor product. The former reaction was catalyzed by Eubacterium L-8 and the latter was by Streptococcus LJ-22. Among GL and its metabolites, GA and GAMG had more potent in vitro anti-platelet aggregation activity than GL. GA also showed the most potent cytotoxicity against tumor cell lines and the potent inhibitory activity on rotavirus infection as well as growth of Helicobacter pylori. GAMG, the minor metabolite of GL, was the sweetest.