Studies On Quality Standards For Morus Nigra Folium Of Xinjiang

Objective: According to the quality standard of Chinese Pharmacopoeia as the guiding principle, using a variety of techniques and methods to establish the methods of character identification, TLC and IR identify, routine inspection of the project is completed, screening of antioxidant active components by Bio-TLC, determination of the content of effective components, determination of heavy metals and harmful elements for Xinjiang medicine mulberry leaf, and on this basis, wrote medicine mulberry leaf quality standard draft. Methods: 1. Using a microscope to identify the microscopic characteristics of mulberry leaves powder. 2. Application of TLC to identify four flavonoids, chlorogenic acid, DNJ, and GABA of drug mulberry leaves. 3. UV spectrophotometry combined with Bio-TLC to study antioxidant active ingredients of medicine mulberry. 4. Using HPLC to establish the methods for the determination of multiple chemical compositions of the drug mulberry leaves, and the QAMS was used to estimate the contents of rutin, Isoquercitrin, chlorogenic acid and astragalin. 5. According to the methods in the general rules of Chinese Pharmacopoeia(2015) to examine moisture content, total ash, acid insoluble ash and alcohol soluble extractives of multiple batches of drug mulberry leaves. 6. IR spectroscopy was used to classify different batches of samples rapidly. 7. The application of ICP-MS method to measure the contents of trace elements in drug mulberry leaves and limited checks of the harmful elements and heavy metals contained in them. Results: 1. Micro structures which are glandular hairs, non glandular hairs, cystoliths, milk ducts and other characteristics are contained in mulberry leaf powder. 2. The optimum chromatographic conditions screened by TLC for the four kinds of flavonoids and chlorogenic acid, DNJ, GABA can well separate these components. 3. The results of antioxidant activity showed that the IC50 of drug mulberry leaves methanol extracts on DPPH, OH?, O2?- are respectively 0.08mg/m L, 13.88mg/m L, 3.93mg/m L. And the rutin, chlorogenic acid, astragalin, Isoquercitrin and quercetin of the methanol extracts are identified owing antioxidant activity by Bio-TLC. 4. Through the methodology study for verification and sample pretreatment methods, the final determination contents are that rutin is not less than 0.3mg/g, chlorogenic acid not less than 0.4mg/g, astragalin not less than 0.4mg/g, Isoquercitrin not less than 0.2mg/g, DNJ not less than 9.0 μg / g and GABA not less than 15.0 μg / g. Using rutin as the standard to establish QAMS method shows that the retention time of chlorogenic acid, Isoquercitrin and astragalin were respectively 0.501, 1.132, 1.591, and concentration relative correction factor respectively 1.700, 1.462, 0.185. There is no significant difference between the results with the QAMS and the external standard method. 5. Moisture contents of different mulberry leaves are between 5.54% and 7.37%, total ash determination between 2.80% and 7.53%, acid insoluble ash measured values between 0.61% and 1.77%, alcohol soluble extractives results between 9.23% and 14.69%. 6. Scanning medicine mulberry leaf powder by infrared spectrum(400-4000cm-1) and selecting ten peaks to make the cluster analysis, the results were that the samples of Kashi, Hotan, qia’er Bagh village can be separated, the samples of Korla and Akesu confused. 7. Heavy metals and harmful elements were lower than the equirement values of Good Agricultural Practise for Chinese Crude Drugs(GAP). Conclusion: 1.In this experiment, according to the requirements of Chinese Pharmacopoeia guidelines to do basic research of Xinjiang medicine mulberry leaves, establish various identification methods such as microscopic characteristics, TLC, IR, routine examination items and content determination, and on the basis written quality standard draft. 2. The combination of activity screening and TLC can not only confirm whether it has antioxidant activity, compared with the Rf of standard substances can also identify the specific antioxidant components. The method is simple and feasible. 3. the contents of effective components calculated by the QAMS and measured through the external standard method were basically the same, so the QAMS method can be used to determine the content of various components under the situation of standard material shortage, especially suitable for the quality control of Chinese herbal medicine index. 4. The ICP-MS method has high sensitivity in the determination of heavy metals, and has various advantages such as wide linear range, simultaneous determination of multi elements and so on.