The Study On Chemical Constituents And Quality Of Woodwardia Unigemmata

This dissertation included two parts: summary of related literatures; study on the chemical constituents of Woodwardia unigemmata from Chendu, establishment of thin liquid chromatography(TLC),the quantitative determination of kaempferol-3-O-α-L-rhamnopyranoside-7-O-α-L-rhamnopyranoside and total flavonoid, chromatographic fingerprint of Gou Ji Guan-Zhong from 13 different habitats by High performance liquid chromatography(HPLC).Part one: The documents about the present research situation in Chinese medicine Gou Ji Guan-Zhong were summed up and analyzed. A review on the research progress of genus Woodwardia in chemical constituents and pharmacological activities was presented.Part two: According to the research on the commercial Guan-Zhong done by my tutor, we choose Woodwardia unigemmata as material to study, because it is one of the most popular plants used as Guan-Zhong while it is not recorded in Chinese pharmacopoeia. In this part, the chemical constituents of Woodwardia unigemmata from Chendu were studied. The qualitative and quantitative analysis of medicinal materials and their effective fractions of Gou Ji Guan-Zhong from different habitats were undertaken respectively.Gou Ji Guan-Zhong samples collected from 13 different habitats were identified. It has become clear that they are from Woodwardia unigemmata and Woodwardia japonica.The chemical constituents of Woodwardia unigemmata from Chengdu were investigated. Three compounds and two mixtures were isolated from the 4 parts of extracted from Woodwardia unigemmata by silica gel, polyamide, sephadex LH-20 column chromatography. On basis of their spectral data, they were elucidated as kaempferol-3-O-α-L-(4-O-acetyl)-rhamnopyranoside-7-O-α-L-rhamnopyranoside, kaempferol-3-O-α-L-rhamnopyranoside-7-O-α-L-rhamnopyranoside andB-sitosterol.kaempferol-3-O-α-L-(4-O-acetyl)-rhamnopyranoside-7-O-α-L-rhamnopyranoside and 8-sitosterol were obtained from Woodwardia unigemmata for the first time. kaempferol-3-O-α-L-(3-O-acetyl)-rhamnopyranoside-7-O-α-L-rhamnopyranoside .kaempferol-3-O-α-L-(2-O-acetyl)-rhamnopyranoside-7-O-α-L -rhamnopyranoside.kaempferol-3-O-α-L-(2,3-O-diacetyl)-rhamnopyranoside-7-O-a-L-rhamnopyranosid-e,kaempferol-3-O-α-L-(2,4-O-diacetyl)-rhamnopyranoside-7-O-α-L-rhamnopyranoside,and kaempferol-3-O-α-L-(3,4-O-diacetyl)-rhamnopyranoside-7-O-a-L-rhamnopyranoside were identified from Woodwardia for the first time.The chromatographic fingerprint of effective fractions of Woodwardia unigemmata from Chengdu was established by HPLC for the first time. Relative retention time and relative area of single peak, which makes up above 3% of the area of all peaks, were determined. In experiments of accuracy and reproducibility, RSD of relative retention time was 3.00-5.18% and 0.15-1.16% respectively, RSD of relative peak area was 2.99-15.29% and 1.52-3.76% respectively. The stability experiment revealed that the sample solution is stable in 8 hours.The chromatographic fingerprint of medicinal materials of Woodwardia unigemmata from Chengdu was also established by HPLC for the first time. Data were analyzed by "SOP of Similarity evaluation system for chromatographic fingerprint of TCM (Version 2004 A) "software to compare the results of accuracy, reproducibility and stability respectively .The results showed that they were of high similarity respectively.Quantitative analysis of total flavonoid in effective fractions and medicinal materials of Woodwardia unigemmata from Chengdu were undertaken, which based on this fact that Kaempferol-3-O-a-L-rhamnopyranoside-7-O-a-L-rhamnopyranoside was used as contrast matter. There was a good linear relationship between ABS of the samples and their concentration (from 0.0014 to 0.0168mg/ml) (r=0.9990), and the average recovery was 94.43% (RSD=0.43%, n=5). The results of accuracy, reproducibility and stability were good.Quantitative determination of Kaempferol-3-0-a-L-rharnnopyranoside-7-0-cc-L-rhamnopyranoside(K390) in effective fractions and medicinal materials of Woodwardia unigemmata from Chengdu were undertaken. Ther...