Studies On Quality Standard Of Schisandrae Sphenantherae Fructus And Quantitative Determination Of Flavonoids In Houttuyniae Herba

1,Nan Wuweizi (Schisandrae Sphenantherae Fructus) are dried ripe fruits of Schisandra sphenanthera Rehd. Et Wils. The types of the chemical consitituents and their contents between Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus were significantly different, but they were both used as one Chinese traditional medicine twenty years ago. Therefore, in Chinese Pharmacopoiea (2000 version), Schisandrae Sphenantherae Fructus were recorded as "Nan Wuweizi" and Schisandrae Chinensis Fructus were recorded as "Wu weizi" respectively. "Nan Wuweizi" recorded by Chinese Pharmacopoeia (VolumeⅠ,2005 version) have included descriptions, TLC identification, HPLC determination, the purity test and other items, yet lack powder microscopic identification, moisture and ash inspection. The TLC identification method which is identical with Schisandrae Chinensis Fructus used deoxyschizandrin as the reference standard. The HPLC chromatographic conditions of determination need to be optimized, and the content limit of schisantherin A in Nan Wuweizi need to be calculated with reference to the dried drug. In addition, Vinegar Prepared Nan Wuweizi recorded in Chinese Pharmacopoeia 2005 include simple processing methods and propertie descriptions. We developed the quality standard of Nan Wuweizi on the base of Chinese Pharmacopoeia 2005. In this paper, properties and powder microscopic identification were studied, a specific TLC identification method use (+)-Anwulignan as standard reference was established, moisture and total ash was added, HPLC determination method was improved and the new content limit was setted for 10 samples of Nan Wuweizi and Vinegar Prepared Nan Wuweizi.A new specific TLC identification method for Nan Wuweizi was established with (+)-Anwulignan and control medicine as chemical reference substance, with 5% phosphomolybdic acid ethanol solvent as chromogenic reagent, developed at 105℃until spots are clear. The spots in the chromatograms obtained with the crude and vinegar prepared drugs of Nan Wuweizi and control medicine were corresponded in position and colour to the spot in chromatogram obtained with the (+)-anwulignan, but no (+)-anwulignan spot was observed in the Wuweizi chromatogram. The results showed that the newly-established TLC method is specific for the qualitative identification of Nan Wuweizi. The moisture and total ash of 10 samples were determined according to the methods in Chinese Pharmacopoeia 2005. According to the testing results, it is proposed to contain less than 12.0% of moisture and 6.0% of total ash calculated to the dried drug.We improve the ratio of mobile phase [THF-H2O (38:62)]of HPLC methods in the base of Chinese Pharmacopoeia 2005 and determined 10 samples' contents of schisantherin A calculated with reference to the dried drug. There was a good linearity (r=0.9999) within the range of 0.0116 mg/ml-0.5800 mg/ml. The average recovery of schisantherin A was 101.7% with RSD of 2.18% (n=6). The contents of schisantherin A were 0.25%～0.74% in Nan Wuweizi. The content of schisantherin A calculated with reference to the dried drug in Nan-Wuweiz were 0.28%～0.80%. It is proposed to contain not less than 0.20% of schisantherin A calculated with reference to the dried drug.We processed 10 Vinegar Prepared Nan Wuweizi samples by the processing methods in Chinese Pharmacopoeia. And we made studies about their shape and properties, powder microscopic identification, TLC identification, HPLC determination, moisture and total ash inspection. The shape and properties of 10 Vinegar Prepared Nan Wuweizi samples are conforms with the regulations of current Pharmacopoiea. There is no significant difference between Vinegar Prepared Nan Wuweizi and Nan Wuweizi. The newly established TLC identification methods for Nan Wuweizi are also applied to Vinegar Prepared Nan Wuweizi, and the results have no significant difference. It is proposed to contain less than 12.0% of moisture and 6.0% of total ash calculated to the dried prepared drug by the testing results. The same HPLC chromatographic conditions were used to determine the content of schisantherin A in Vinegar Prepared Nan Wuweizi, and it is proposed to contain not less than 0.20% of schisantherin A calculated with reference to the dried prepared drugs.2,Yuxingcao(Houttuyniae Herba) are fresh or dried aerial parts of whole plant of Houttuynia cordata Thunb. It tranditionally used to remove toxic heat, promote drainage of pus, and relieve dysuria. As a tradintional Chinese medicine for heat-clearing and detoxication, Yuxingcao is also officially identified as a drug-diet herb by the Ministry of Health. Studies by our group showed that anti-complement activity is one of the mechanisms in playing heat-clearing and detoxication efficacy by traditional Chinese medicine. Jiang and Zhang had carried out chemical studies of herbs and anti-complement activity of its compound. The results showed that the flavonoids (such as quercitrin, isoquercitrin, etc.) have anti-complementary activity, as well as flavonoids have anti-virus, anti-inflammatory, anti-allergy activity reported in the literature. Therefore these flavonoids can be used as markers for quality control of Herba Houttuyniae. The cotents of total flavone of 11 samples were determined by UV-VIS method. LC-MS/MS method was used to analyze chemical composition of 50% methanol extract solution. As well as HPLC method was used to determine the major flavonoids (rutin, hyperoside, isoquercitrin, quercitrin) of 11 samples from different collection. The results showed that content of flavonoids in Houttuynia cordata Thunb. are significantly different.The contents of total flavone are in the range of 12.897 70.352 mg/g. The contents of flavonoids in fresh sample are much higher than dried sample calculated with reference to the dried drug. And at different parts of fresh sample, the contents of flavonoids are highest in leaves, lower in stems and the lowest in stems and roots, in stems and roots only trace amounts of hyperoside was detected, the other three flavonoids were not detected. The results provide a basis for the quality control and reasonable application of Houttuyniae Herba.