| Objective:Accroding to modern analysis technology and pharmacognostical methods, authenticity identification,fingerprints,effective parts and components contents of Rhizoma Smilacis Glabrae were studied to establish the mode of HPLC finger-print and quality evaluation system and to provide a further scientific basis to normalize the application and quality control.Method:1.Character and microscopic identification methods were performed to qualitatively identify Rhizoma Smilacis Glabrae by ChP.2010.2.The process of extracting effective total flavonoid from Rhizoma Smilacis Glabrae was optimized by orthogonal experimental design. The content of total flavonoid was determined by ultraviolet spectrophotometry. The control article was Rutoside.The developer was sodium nitrite-aluminum nitrate. The content of total flavonoid was determined at 500nm.3.The process of extracting effective constituents from Rhizoma Smilacis Glabrae was optimized by orthogonal experimental design. The content of Taxifolin,Astilbin and Resveratrol was determined by RP-HPLC. The determination was performed on a Kromasil C18 (250×4.6mm,5μm) column.The mobile phase was methanol-0.5%phosphonic acid solution(34:66).UV detection wavelength was at range of 200-800nm. The flow rate was 1mL-min-1 and the column temperature was at 30℃.4.The HPLC finger-print was studied by RP-HPLC. The determination was performed on a Kromasil C18(250×4.6mm,5μm)column. The mobile phase was methanol-0.5%phosphonic acid solution by gradient elution. UV detection wavelength was at 290nm.The flow rate was 0.8 mL·min-1 and the column temperature was at room temperature.Results:1.Observations show that the characteristics were roughly similar in samples 1 to 12, which accord with the morphological identification features in ChP.2010.Vol I..The samples 13 and 14 were non-compliance.The result of starch granules observations show that the morphological characterisitics of samples 1 to 13 were roughly similar,which accord with ChP.2010.Vol I..The sample 14 was non-compliance.2.The parameter of optimum extraction process of total flavonids by orthogonal experimental design should be 50% alcohol with 50 times amount of materials, 1 h every time.The total flavonoids content from different samples was in the range of 2.9%-18.7%.The sample from Vietnan Henoi was the highest level, the sample from Huazhou of Huazhou of Guangdong province are the lowest.The content of the slices from medicinal materials companies were higher levels and closer, The quality goods from different producing areas relatively significant, the content of two pseudo-products were lower.3.The parameter of optimum extraction process of taxifolin,astilbin and resveratrol extraction by orthogonal experimental design should be 60% alcohol with 30 times amount of materials,35min every time.The result of effective constituents content determination showed that the content of taxifolin was between 0.00869% and 0.06996%.The samples of Hunan,Shangsi of Guangxi province,Hanoi were below the limitation standard.The content of astilbin was in the range of 0.19669%-4.90443%.The samples of Hunan,Hanoi,Huazhou were less than 0.45% stipulated in ChP.2010;the content of resveratrol was in the range of 0.0137%-0.1551%.4.Make similarity evaluation of all the samples by MATLAB.According to the results,the fingerprints similarity of population was lower,in which samples from closer region, Guangdong Luoding hospital and Hongkong,were higher,but the similarity of the two samples collected from Guangxi and the two pseudo-products were lower.Through principal component analysis,the six samples can be divided into two categories,mode I and II.Conclusion:1. Accroding to ChP.2010.Vol I, character and microscopic identification methods were performed to qualitatively identify Rhizoma Smilacis Glabrae from 14 samples. Character identification showed that the samples from Hu Nan and Vietnan Henoi didn't accord with Chinese Pharmacopoeia. Microscopic identification showed the sample from Vietnan Henoi didn't accord with Chinese Pharmacopoeia.According to macroscopic identification,and could be detemined counterfeit product.But the features of starch grain accord with Chinese Pharmacopoeia, so it could be auxiliary basis, but not judging paternity..2.The process of extacting total flavonoid from Rhizoma Smilacis Glabrae was optimized by orthogonal experimental design.Make quantitative analysis method of total flavonoid and determine the content of total flavonoid from 14 samples.The results showed the content of total flavonoid from 14 samples were between 2.9%-18.7% and relatively significant.The contents of total flavonoid in the market were higher and the pseudo-product are lower.3. The process of extacting Taxifolion,Astilbin and Resveratrol from Rhizoma Smilacis Glabrae was optimized by orthogonal experimental design.Make quantitative analysis method of effective components and determine the content of effective components from 14 samples.The results show the content of Taxifoliol from 14 samples are between 0.00869% and 0.06996%.The samples from Hu Nan,Guangxi Shangsi and Vietnan Henoi are below the limitation standard;The content of astilbin are between 0.19669%-4.90443%.The samples from Hu Nan,Vietnan Henoi and Guangdong Huazhou are less than 0.45% stipulated in ChP.2010. The contents of Resveratrol are in the range of 0.0137%-0.1551%.The effective component content of 2 pseudo-products were lower, Taxifolion of all were below the limitation standard, Astilbin were less than 0.45%, Resveratrol below the limitation standard 0.22%. 4.Established the mode of HPLC finger-print of Rhizoma Smilacis Glabrae By identification similarity assessment and principal component analysis, built quality control method comprehensively by MATLAB.Accroding to the results,the samples according with HPLC fingerprint Class I were better and it consistented with results of the component content study.5.According to ChP.2010,the standard of quality control was improved to Rhizoma Smilacis Glabra.5.1 Characteristics identification:The section is light brown,brown or almost white.5.2Microscopical identification:The powder of Rhizoma Smilacis Glabra are light brown,almost white with lots of starch grain.Those single starch grains were almost round,polygonal shape and square shape with diameter of 8-50μm. The centre of starch grain were sutural,stellate,trigeminal shape,tetrageminal shape or spot.The bigger starch grain has texture gradient. Compound starch grains were composed of 2-4 single ones.5.3Content:Calculated on dry matters, the contents of total flavonoid weren't less than 4%,Taxifolin weren't less than 0.01%, Astilbin weren't less than 0.45%, Resveratrol weren't less than 0.22%.
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