Studies On The Extraction And Purification Technology And Quality Of Woodwardia Unigemmata Effective Fractions

This dissertation included two parts: summary of related literatures; study on the extraction and purification technology and quality of Woodwardia unigemmata effective fractions.Part one:A review on the development of anti-influenza herbal drugs for treatment and prophylaxis; A review on the research progress of genus Woodwardia in chemical constituents and pharmacological activities was presented.Part two:Studies on the extraction and purification technology of Woodwardia unigemmata effective fractions.using L9(34) orthogonal test which were evalued by the extraction efficiency of total flavones and indicative constituents kaempferol. Give the result that the best extracted term is that the drug is extracted by A2% alcohol for twrice, two hours each time,and the quantity of solvent is 10 times of drugs.To study the purification technology of Woodwardia unigemmata effective fractions.After investigated the adsorptive property of 6 kinds of macroporous adsorptive resins,A and D resins was selected due to its best adsorptive and separative property of total flavones.We investigated the loading capacity, the ratio of diameter and height, concentration of obsorptive solution, absorptive rate, purified solvent,velocity of flow, eluting solvent and the repeated using times of resins,and et al. The optimized purification technology was accept.The extraction and purification technology discribed above was tested and verified through 3 batches of samples of effective fractions.The preparation and determination results showed that this extraction and purification technology was steady and satisfied, the yield ratio of effective fractions was 1.27%, the content of total flavones was 64.77%, the transferring ratio was beyond 51%. Studies on the chemical constituents of Woodwardia unigemmata effective fractions.Several separation and purification methods and determination technology(such as UV, EI-MS, ESI-MS, 1H-NMR, 13C-NMR, HMBC) were used,two compounds were separated and identified from it.They are kaempferol-3-O-α-L-rhamnopyranoside-7-O-α-L-rhamnopyranoside and kaempferol-3-O-α-L-(4-O-acetyl)-rhamnopyranoside-7-O-α-L-rhamnopyranoside.Establishment of the quanlity controlling methods of Woodwardia unigemmata effective fractions. The quantitative methods of indicative constituents:kaempferol-3-O-α-L-(4-O-acetyl)-rhamnopyranoside-7-O-α-L-rhamnopyranoside by HPLC was established.The linear regression was :A= 58464x + 34.5,r=0.9999 , the average recoveries was 101.62 %,RSD =1.52 % (n = 6).The determination results in three batches of the effective fractions showed that the methods were steady and reliable and can be used as the determination methods.Establish methods of determinating the total flavones in the effective fractions of Woodwardia unigemmata. The quantitative methods of indicative constituents:kaempferol by HPLC was established. The linear regression was :y = 3882x - 4.364 ,r = 1 , the average recoveries was 98.66 %,RSD =1.17 %(n = 6). The determination results in three batches of the effective fractions and raw drugs showed that the methods were steady and reliable and can be used as the determination methods.The chromatographic fingerprint of effective fractions of Woodwardia unigemmata effective fractions was established by HPLC. Relative retention time and relative area of single peak, which makes up above 5 % of the area of all peaks, were determined. In experiments of accuracy and reproducibility, RSD of relative retention time was 0.20-1.93 % and 0.23-2.13% respectively, RSD of relative peak area was 2.42-3.46 % and 2.19-3.48 % respectively. The stability experiment revealed that the sample solution is stable in 24 hours.