| Objective:As one of the Traditional Chinese Medicine (TCM) Psammosilene tunicoides W. C. Wu et C. Y. Wu is commonly used in clinic, which is the only species growing in the south-west of China. Psammosilenes Radix is a famous Miao national Herb, widely used in the clinical medicine, which has the effect of Qufeng desiccant, relieving pain and swelling detoxification. At the moment, most of researches about the P. tunicoides are mainly focused on the chemical composition and pharmacological effects. Based on the related references, the triterpenoid saponins are major chemical constituents of the Psammosilenes Radix, in addition, it also containing peptides, amino acid, organic acid and so on. New results from the pharmacology researches reveal that the triterpenoid saponins of the Psammosilenes Radix have analgesic and anti-inflammatory effects. It was also found that quillaic acid mainly existed in the aglycone of Psammosilenes Radix. In recent years, the studies on the components and pharmacological effects of the Psammosilenes Radix have increased, while less research about the methods of quality control. Chinese Pharmacopoeia 2010 only contains identification but without quantitative control. After the tests for the quality, the extractives are often unqualified. So improvement of the quality standards of the Psammosilenes Radix was ergent and meaniful.Methods:(1) The quality standards were established according to 10 batches of Psammosilenes Radix. Microscopical characteristics were studied for transverse section.(2) The extracts of the Psammosilenes Radix were determined by methods described in appendices of Chinese Pharmacopoeia.(3) The quillaic acid was determined by HPLC. The temperature of Agilent Zorbax Eclipse Plus C18 (250 mm×4.6 mm,5 μm) chromatogram column was 25℃, the mobile phase was water-acetonitrile. And the flow rate was 1 mL·min-1 and the detection wave-length was 210 run, and we have made a reasonable revision on the content limits.(4) In the finger-print research, the technique of gradient elution was adopted in HPLC and HPLC-MS, the results of the relative retention time (RRT) and relative peak area (RPA) were evaluated with the medicine chromatographic finger-print similarity evaluation software and the SPSS software.Results:(1) We established the standard of 10 batches Psammosilene tunicoides W. C. Wu et C. Y. Wu transverse section identification. Microscopic identification can provide an effective method in identification of TCM. Parts of the cork layers remained, the phelloderm had several multi-column cells which were arranged tangentially, the fissures was existed in the phelloderm. The phloem was narrow; instead, the xylem was wide. The cambium presented ring-shaped between the phelloderm and phloem.(2) The item of extract was optimized. According to the method of Chinese Pharmacopoeia 2010 and considering the active ingredients, we built up a suitable method of extractives by using the cold soak, hot soak and different solvents including different solvent concentrations. According to the average results of the extractives, we suggest that the extractiveis of the herb should contain more than 30% by using hot soak with the 80% ethanol.(3) We established a method for content determination. Calculated on dry product, containing quillaic acid should not less than 0.10%. We also has a research on the extraction method of sample. L9(34) orthogonal experiment was adopted, the total saponins acid hydrolystate effects on concentration of acid, reaction time and reaction temperature.(4) The finger-print of the total saponins acid hydrolysate was established. The finger-print spectrum was evaluated with the software "Similarity Evaluation System for Chromatographic Fingerprint of TCM (version 2004A)". According to the results of clustering analysis, there was a difference between the wild samples and the cultivated ones. The results of similarity and cluster were consistent:the cultivated tunicoides with thin roots were clustered into one group; the cultivation herb with thick roots and the wild ones were classified into the same cluster.Conclusion:In this paper, we improved and established standards of Psammosilene tunicoides-- transverse section, extracts, assay and finger-print. The methods were simple, accurate; these methods could also support the further improvement of the Psammosilenes Radix quality standards.
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