Research On Quality Standards Of Herba Humuli Scandens And The Active Component For The Inhibition On Mycobacterium Tuberculosisin

To coordinate with the Sichuan scientific and technological project to improve the quality standard of the local conventional medicines in Sichuan Province, This research systematically studied the measures of quality control of Herba Humuli Scandens and developed the draft of quality standards including the source, characters, identification (microscopic identification,TLC), checking(waters,total ash,acid insoluble ash), determination of extractives and the content determination. Some of them have been included in the "standard of Chinese herbal medicines in Sichuan Province"2008 edition. In addition, the active component for the inhibition on Mycobacterium tuber culosisin of Herba Humuli Scandens was studied for the further development and utilization.Main contents and results are as follows:1.This is the first detailed study on the morphological organization of Herba Humuli Scandens to identify its original plant,morphological characters and organizational structure. Surface of stem and petiole barbed spines, leaf shape, leaf surface coat conditions, glandular dots, male and female inflorescence shape, the color of perianth, ovary, stigma, and other aspects of fruit can be identified from the Original plant. The color, barbed spines and pilose on the stem surface, the shape of leaves, the conditions of leaf surface coat, glandular dots, barbed spines on the petiole and other aspects can be identified from the Herbs. The surface coat, outer phloem fiber bundles and the proportion of each part can be identified from the stem cross-section. The upper and lower surface coat, leaf vascular tissue differentiation and other features in the veins can be identified from the cross-section of leaf veins. The fibers, non-glandular hairs, glandular hairs, the crystal cells containing limestone, calcium oxalate crystals can be identified with microscopy from the powder.2.The TLC method of Herba Humuli Scandens was established. The good separation was taken by using the toluence-ethyl acetate-formic acid (8:2:0.5) as the mobile phase. The Rf values were moderate and the chromatographic spots were clear. The TLC tests of ten batches of commercial medicinal materials and local medicinal materials from different origins showed that this method was adaptable and reproducible in the identification of luteolin in Herba Humuli Scandens.3.The moisture, total ash and acid insoluble ash of ten batches of commercial medicinal materials and local medicinal materials from different origins were checked by the methods from the "Chinese Pharmacopoeia" 2010edition. According to the measured data and the actual situation of production and trade, The limit was developed as the follows:water, total ash and acid insoluble ash could not be over 10.0%, 22.0% and 6.0%, respectively.4. The hot dipping method with 75% ethanol to determine the extract from Herba Humuli Scandens were established. Based on the measured data of 10 batches of samples and the actual situation of production and trade, the limit of extract content was set as not less than 12.0%.5. The HPLC method to determine the luteolin in Herba Humuli Scandens was established. It is accurate, precise, simple, fast and easy to operate. Based on the measured data of 10 batches of samples and the actual situation of production and trade, the limit of luteolin content was set as not less than 2.0%.6. The active component for the inhibition on Mycobacterium tuberculosisin was firstly studied. The active component in Herba Humuli Scandens for the inhibition on Mycobacterium tuberculosis was screened on different culture medium. The ethyl acetate has obvious inhibitory effect, and the minimum inhibitory concentration is 31.25mg/ml. According to the results of in vitro screening, the chemical composition of the effective area was isolated, and structure was identified. Four compounds were isolated from the ethyl acetate extract. With the physical and chemical constants determination and analysis of 1H-NMR,13C-NMR spectroscopy, they were identified asβ-sitosterol, luteolin,β-daucosterol and apigenin-7-O-β-D-glucosidase. The apigenin-7-O-β-D-glucosidase was firstly isolated from this plant.