| Ilex rotunda Thunb., which has other names as longdanzai, baimuxiang, baichenxiang, misuimu, shandongqing, baipidongqing, dayedongqing, baishanye, et al. It’s from the cortex and velamen of Ilex rotunda Thunb., bitter taste and cold-natured, which has been a remedy for clearing heat and removing toxicity, removing dampness through diuresis, promoting the circulation of qi, relieving pain and hemostasis in Southern China. Modern pharmacology study showed that it possessed actions as cold, tonsillitis, pharyngolaryngitis, acute gastroenteritis, dysentery and ostalgia.As a traditional Chinese medicine, Ilex rotunda Thunb. has been recorded in Lingnan Cai Yao Lu, it said that Baimuxiang, bitter taste and clear the heat poison. It also has been recorded in Chinese Pharmacology(1977and2010Edition), the quality of Ilex rotunda Thunb. are mostly controlled by medicinal material properties identification, powder microscopic identification, TLC contrasted with standard medicine and HPLC contrasted with representative chemical composition. But these control method is unilateral, can not set the quality standards for chemical composition of Ilex rotunda Thunb. systematically. In addition, not only cortex and velamen of Ilex rotunda Thunb. combine on the market, but also has large differencens of thickness and quality, even combine with its related species, influence the clinical standard and effects of Ilex rotunda Thunb.Therefore, systematic separation and identification were studied on this medicine. On the basis, we screened suitable index components to establish quality evaluation method using HPTLC chromatographic fingerprint in the material medicine, and compare the differences of HPTLC between different position of Ilex rotunda Thunb. and its related species, which may help to establish the quality standard of this medicine. At the same time, Ilex rotunda Thunb. has large content of saponins and most of them are the effective constituent, just like peduncu-loside. Our study used anatomical, histochemical methods to investigate the saponins localization in Ilex rotunda Thunb., it may help the rational exploitation and application of Ilex rotunda Thunb., also improve the study of Ilex rotunda Thunb.The results showed as follows:1. The determination method of HPTLC fingerprinting of Ilex rotunda Thunb.. The fingerprint characteristics have been constructed by eleven stable peaks of thirteen batches drug materials collected from different areas. Comparing with reference substances, we could identify syringin, peduncu-loside and rotundic acid, respectively. The fingerprint characteristics have been obtained through " ChromafingerTM2005" solution software, and authentication and quality assessment have been analyzed by similarity analysis. The similarities of samples are all above0.9, except one sample is0.84, there have bigger thickness of rinds in that sample, which is easily recognized from other samples. HPTLC for determination the fingerprinting of Ilex rotunda Thunb. is simple, systematic, specificity, and is beneficial for identification of different plant plant parts and quality control of the material medicine.2. HPTLC fingerprinting of Ilex rotunda Thunb. was used to compare the differences between different thickness of Ilex rotunda Thunb., at the same time to compare the differehces between different positions(including bark, velamen, core of stem and leaf) and its related species. The result shows that there are notable differences between different position and different thickness. Compare with the common pattern, the samples with thickness from0.5mm to2mm have the highest similarity, samples with thickness from2mm to7mm take second place. But sample with thickness from9mm to12mm can be easily differentiated from other samples, syringin and peduncu-loside have no corresponding spots in this sample, the position and lightness of other spots are all different from other samples. In addition, as the thickness increases, the lightness of syringin spot would trail away, the result explain that thickness have an affect on content of chemical composition.The chromatogram of bark has the highest similarity with common pattern; velamen has less content of syringin because the spot is not obvious; all the spots of the core are have less lightness, the chromatogram of leaf has the lowest similarity with common pattern, the content of syringin and peduneu-loside are less than other positions obviously. The other relatives can be easily differentiated from Ilex rotunda too. They can not be used as Ilex rotunda Thunb.3. Anatomical and histochemical methods were used to investigate the saponins localization in Ilex rotunda Thunb. Use5%vanillin-glacial acetic acid and perchloric acid as derivatization reagent to dye different cross-section of Ilex rotunda Thunb., The negative control product were the biopsy tissue which socked with FAA for1months. Histochemical localization results showed that saponins accumulated mainly in parenchyma cell of vegetative organ of Ilex rotunda Thunb., saponins distributed in secondary phloem and cortex of root, in secondary phloem and phloem of stem, mainly in palisade tissue of leaf.In this paper, establishing chromatographic fingerprint in the material medicine can be used to identify and evaluate the quality of Ilex rotunda Thunb.. The result showed that different parts or different thickness may affect the quality of the medicine, The other relatives can be easily differentiated from Ilex rotunda Thunb. too. They can not be used as Ilex rotunda Thunb.. At the same time, our study used anatomical, histochemical methods to investigate the saponins localization in Ilex rotunda Thunb. |
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