• 생약학회지
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• 제37권1호통권144호
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• pp.33-36
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• 2006

The quantitative analytical method for the content of dioscin in Dioscorea rhizoma using a high performance liquid chromatography(HPLC) system was established for its quality control. After extracting Discorea rhizoma flour with ethanol, dioscin was separated though an ODS column with a flow rate of 1.0ml/min of acetonnitril-methanol-0.01 M phosphate buffer (pH 5.0)(6:3:1, v/v%), and the amount of dioscin was detected at 210nm. Relation time of dioscin in HPLC chromatogram were about $7.45{\pm}0.34\;min$ and calibration curve showed good linearity at concentrations from 0.1 to 2.0 mg/ml of dioscin. When 5 commercial Dioscorea rhizoma flours were analyzed by HPLC, the peak for dioscin was well separated from others in ethanol extract of Dioscorea rhizoma, and the amount of dioscin was in the range from 0.076 to 0.142(w/w)%.

• 생약학회지
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• 제32권2호통권125호
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• pp.153-156
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• 2001

The methods for isolation and quantitative determination of dioscin, a spirostanol triglycoside, from the roots of Smilax china have been developed. Isolation of dioscin was achieved by silica gel and RP-18 chromatography. The HPLC method used for quantitative determination of dioscin enabled the standardization of the crude drug. It suggests that the content of dioscin in Smilax china should be above 0.01%.

• Mass Spectrometry Letters
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• 제4권3호
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• pp.55-58
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• 2013

Dioscin is a biologically active steroidal saponin with anticancer and hepatoprotective effects. A rapid, selective, and sensitive liquid chromatographic method with electrospray ionization tandem mass spectrometry was developed for the quantification of dioscin in rat plasma. Dioscin was extracted from rat plasma using ethyl acetate at acidic pH. The analytes were separated on a Halo C18 column using gradient elution of acetonitrile and 0.1% formic acid and detected by tandem mass spectrometry in selected reaction monitoring mode. The standard curve was linear ($r^2$ = 0.998) over the concentration range of 1-100 ng/mL. The lower limit of quantification was 1.0 ng/mL using 50 ${\mu}L$ of plasma sample. The coefficient of variation and relative error for intra- and inter-assay at four QC levels were 1.3 to 8.0% and -5.4 to 10.0%, respectively. This method was applied successfully to the pharmacokinetic study of dioscin after oral administration of dioscin at a dose of 29.2 mg/kg in male Sprague-Dawley rats.

• 생약학회지
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• 제20권2호
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• pp.76-82
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• 1989

Five known diosgenin glycosides have been isolated from the MeOH extract of Smilax china rhizomes and characterized as prosapogenin A of dioscin, dioscin, gracillin, methyl protogracillin, methyl protodioscin and its corresponding 22-hydroxy analog. This is the first isolation of the former four compounds from this plant. ${\beta}-Sitosterol\;glucoside$ was also isolated and identified.

• Archives of Pharmacal Research
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• 제17권4호
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• pp.218-222
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• 1994

The natural product, spirostanol glycoside dioscin, was shown to directly inactivate human pleural fluid phospholipase $A_2{\;}(PLA_2)$ Inactivation was dose, and time dependent. The $IC_{50}$ was estimated at 18 .mu.M and virtually complete inactivation of the enzyme occurred at 50 .mu.M. Using Michaelis-Menten kinetics, dioscin inactivated the enzyme by a competitive inhibitory manner, the apparent Ki value was $6.9{\times}10_{-4}$. Reversibility was studied directly by dialysis method, the inhibition was reversible. Additioin of excess $Ca^{2+}$ concentration up to 8 mM did not antagonize the inhibitory activity of dioscin. Inactivation of several kinds of $PLA_2$ by dioscin is due to interaction with the active site of $PLA_2$ and may be a useful adjunt in the theraphy of inflammatory diseases.

• 한국자원식물학회지
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• 제21권6호
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• pp.444-448
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• 2008

An adventitious root formation protocol from Smilax china L. was established for in vitro production of dioscin, a steroidal saponin having various bioactivities such as anticancer, antifungal, antiviral, and antiobesity. Optimal medium for root initiation from leaf explant was MS medium containing $30\;g{\cdot}L^{-1}$ of sucrose supplemented with $1.0\;mg{\cdot}L^{-1}$ kinetin + $2.0\;mg{\cdot}L^{-1}$ NAA. The induction of adventitious roots from in vitro initiated root segments was most favorable to MS liquid medium with $0.1\;mg{\cdot}L^{-1}$ kinetin + $2.0\;mg{\cdot}L^{-1}$ NAA. Among the 20 different adventitious roots originated from different plants, strain No. 10 was selected based on production ability of dioscin, and its stability through the successive suspension culture. The maximum growth stage of adventitious roots was noticed at 5 weeks after subculture while that of dioscin production in the adventitious root was at 7 weeks after subculture in suspension culture system. These results provide that suspension culture of adventitious roots of Smilax china L. have a potential for in vitro mass production of dioscin.

• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.1011-1017
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• 2010

A novel dioscin-glycosidase that specifically hydrolyzes multi-glycosides, such as 3-O-${\alpha}$-L-($1{\to}4$)-rhamnoside, 3-O-${\alpha}$-L-($1{\to}2$)-rhamnoside, 3-O-${\alpha}$-L-($1{\to}4$)-arabinoside, and ${\beta}$-D-glucoside, on diosgenin was isolated from the Absidia sp.d38 strain, purified, and characterized. The molecular mass of the new dioscin-glycosidase is about 55 kDa based on SDS-PAGE. The dioscin-glycosidase gradually hydrolyzes either 3-O-${\alpha}$-L-($1{\to}4$)-Rha or 3-O-${\alpha}$-L-($1{\to}2$)-Rha from dioscin into 3-O-${\alpha}$-L-Rha-${\beta}$-D-Glc-diosgenin, further rapidly hydrolyzes the other ${\alpha}$-L-Rha from 3-O-${\alpha}$-L-Rha-${\beta}$-D-Glc-diosgenin into the main intermediate products of 3-O-${\beta}$-D-Glc-diosgenin, and subsequently hydrolyzes these intermediate products into aglycone as the final product. The enzyme also gradually hydrolyzes 3-O-${\alpha}$-L-($1{\to}4$)-arabinoside, 3-O-${\alpha}$-L-($1{\to}2$)-rhamnoside, and ${\beta}$-D-glucoside from [3-O-${\alpha}$-L-($1{\to}4$)-Ara, 3-O-${\alpha}$-L-($1{\to}4$)-Rha]-${\beta}$-D-Glc-diosgenin into diosgenin as the final product, exhibiting significant differences from previously reported glycosidases. The optimal temperature and pH for the new dioscin-glycosidase is $40^{\circ}C$ and 5.0, respectively. Whereas the activity of the new dioscin-glycosidase was not affected by $Na^+$, $K^+$, and $Mg^{2+}$ ions, it was significantly inhibited by $Cu^{2+}$ and $Hg^{2+}$ ions, and slightly affected by $Ca^{2+}$ ions.

• BMB Reports
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• 제40권6호
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• pp.1016-1020
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• 2007

In this study, dioscin was isolated from Dioscoreae Rhizoma (DR), which is the rhizome of Dioscorea batatas DECNE. that inhabits broad areas of Korea and Japan. To determine whether dioscin induced growth hormone (GH) release, we evaluated its induction effects on GH release both in vitro and in vivo. The 70% methanol extract of DR, and its n-hexane and n-BuOH fractions, induced rat GH (rGH) release in rat pituitary cells 10-fold, 8-fold, and 5-fold higher than the control ($0.36{\pm}0.02 nM$), respectively (p < 0.05 each). The dioscin-induced rGH release of the cells was concentration-dependent and its $ED_{50}$ was $1.14{\times}10^{-5} M$. Within 90 minutes after intravenous administration of $10{\mu}g$/kg (p < 0.05 at $t_{max}$), dioscin caused the greatest increase in rGH concentration ($C_{max}$) in the rat plasma ($34.16{\pm}14.10 ng/ml$) (n = 4), which was twice as high as the control group ($12.88{\pm}3.29 ng/ml$) (n = 27).

• Journal of Plant Biotechnology
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• 제32권3호
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• pp.217-223
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• 2005

액체배양을 통한 부채마의 부정근 배양시 첨가되는 생장 조절제의 종류와 농도에 따른 부정근의 생장과 steroidal saponin 생산과의 관계를 조사하였다. NAA와 BA 농도별 부정근 생장은 NAA를 단독으로 1.0 mg/L를 처리한 곳에서 가장 효과적이었으나 NAA에 BA를 첨가하면 부정근의 생장에 오히려 억제효과를 보였다. 20종의 각각 다른 배양주로부터 dioscin, prosapogenin A 및 prosapogenin C의 함량을 조사한 결과 배양주에 따라 큰 차이를 보였는데, 그 중 dioscin 함량이 건물당 2.5%로 가장 높았던 10번 배양주를 선발하였다. 부정근에 함유된 세 종류의 steroidal saponin 함량은 부정근의 생장이 왕성한 4주까지는 낮으나 부정근의 생장이 거의 정지되는 5주째에 급격히 증가하였다. Dioscin과 prosapogenin C의 함량은 NAA보다 IBA를 처리한곳에서 현저히 높으나 prosapogenin A의 함량은 이들 두성분의 종류와 농도에 영향을 받지 않았다.

• 생약학회지
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• 제24권3호
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• pp.187-191
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• 1993

Three compounds were isolated from the rhizomes of Dioscorea quinqueloba and identified as $diosgenin\;3-O-{\alpha}_-L-rhamnopyranosyl(1{\rightarrow}2)-{\beta}-_D-glucopyranoside$, dioscin and gracillin by chemical and spectroscopic means.