Research Of The Quality Control Method Of The Traditional Chinese Medicine Liuweidihuang Pills

Liuweidihuang Pills is a classical recipe in nourishing the kidney and it is widely used for the insufficiency of kidne-yin. It plays a very important role in the traditional Chinese medicines. In the Chinese pharmacopoeia 2000, the minister Fructus Corni and the assistant Cortex Moutan are determined. There is no quality control method of the monarch Radix Rehmanniae Preparata. The ursolic acid in Fructus Corni is determined by thin layer chromatography scanning method and the paeonol in Cortex Moutan is determined by ultraviolet spectrometry after distillation. It seems too complicated to determine the two constituents in the minister and the assistant by two methods. The purpose of this research is: first to search the quality index of Liuweidihuang Pills and establish its quality control method; second to improve the quality control method of Fructus Corni and Cortex Moutan in the Chinese pharmacopoeia 2000. To gain a comprehensive analysis result, we employed the high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) and the capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) methods in the determination of Liuweidihuang Pills, Cortex Moutan, Fructus Corni, Radix Rehmanniae Preparata and Rhizoma Alismatis. 37 constituents including ursolic acid and paeonol were identified by the fragments of mass spectrometry.The major constituents in Liuweidihuang Pills and its composed herbs were determined by capillary electrophoresis in Chapter 2. The capillary electrophoresis quantitative methods were established for the determination of paeonol, ursolic acid, gallic acid, paeoniflorin, morroniside and loganin in Liuweidihuang Pills for the first time. The analytical conditions for the simultaneous determination of four constituents paeonol, paeoniflorin, morroniside and loganin in Liuweidihuang Pills were optimized. To search the quality index of the monarch, we roasted the Radix Rehmanniae Preparata sample and for the first time established a capillary electrophoresis method for the determination of catalpol content in the Radix Rehmanniae Preparata sample at different times. The catalpol is known to have a pharmacological effects of activating urination, lowering blood sugar level and purgation by demulcents, so it is not suitable to be used as the quality index of the Radix Rehmanniae Preparata. While in this analysis we found a strongly absorbed peak (210 nm) at the retention time of 9.8 min. It is well separated under a buffer conditon of 20-80 mmol L-1 borate solution.The unknown peak was prepared by thin layer chromatography in Chapter 3. The results of the structure identification show it is dihydromelittoside. We established a capillary electrophoresis quantitative method for the determination of dihydromelittoside in the Radix Rehmanniae Preparata and the Liuweidihuang Pills for the first time. The pharmacological activity of dihydromelittoside was investigated for the first time to see whether the pharmacological activity of this compound was in relation to the monarch Radix Rehmanniae Preparata and the Liuweidihuang Pills. The pharmacological experiment was parallelly carried out using dihydromelittoside, the Radix Rehmanniae Preparata and the Liuweidihuang Pills at organize level and sub-cell level. The LPO amount and the SOD activity were used as the pharmacological index. The results of the pharmacological experiments show that dihydromelittoside has an effect of cleanup free radicals and enhances the activtity of SOD. These effects are in accordance with Liuweidihuang Pills, so dihydromelittoside is suitable to be used as the quality index of the monarch Radix Rehmanniae Preparata.The high-performance liquid chromatography and the capillary electrophoresis methods with ultraviolet detection have some difficulty in the qualitative analysis of the traditional Chinese medicine. The mass spectrometer as the detector can offer rich information of the fragments. The Liuweidihuang Pills, Cortex Moutan, Fructus Corni, Radix...