Analysis And Pharmacokinetics Study On Ingredients In Chinese Complex Prescription Gegen Qinlian Decoction

Chinese complex prescription Gegen Qinlian Decoction (GQD), consisted of Pueraria, Scutellaria, Coptis Rhizome and Glycyrrhizae Radix, was firstly described in"Treatise on Febrile Diseases". It can be used to relieve muscles and fever, stop diarrhea. Because it contains less ingredients and the effects is clear, GQD is often used as a classic prescription by the traditional Chinese physicians. The platforms of high performance liquid chromatography (HPLC), two-dimensional liquid chromatography coupled to mass spectrometry (2D-LC /MS) and pressurized capillary electrochromatography (pCEC) were established for the separation and analysis of effective ingredients in GQD, such as flavonoids, alkaloids and etc.. Using Berberine and Baicalin as main indexes and HPLC as analytical tool, the pharmacokinetics of GQD in rat was investigated. The detailed results of the investigation were presented as follows.1. A system of two-dimensional liquid chromatography coupled to mass spectrometry (2D-LC /MS) was developed for the separation and analysis of ingredients in GQD. An CN column was used in the first dimensional chromatography, with gradient elution using water and methanol as the mobile phase A and mobile phase B, respectively; An ODS column was used in the second dimension with gradient elution using 20 mmol·L-1 ammonium acetate buffer-acetonitrile as the mobile phase A and mobile phase B, respectively. The effluent was detected by mass spectrometry with ESI/APCI combined ion source, in both positive and negative scan modes. Compared with UV absorption detection, MS detected more components, especially in the negative mode. The 2D-LC /MS system provides higher peak capacity, better resolution and the capacity of peak identification. It is suitable for the analysis of complex samples such as the Chinese complex prescriptions.2. A micro-column analytical method, pressurized capillary electrochromatography (pCEC) was also developed for the separation of ingredients in GQD. pCEC, which combines micro high performance liquid chromatography (micro-HPLC) and caplillary electrophoresis (CE), provides high efficiency, high selectivity and high resolution. Five effective ingredients in GQD including Jatrorrhizine, Palmatine, Berberine, Baicalin and Glycyrrhizic acid were separated on a capillary column packed with 5μm C18, by using a mobile phase of methanol-0.5% TEA. The separation was optimized with respect to pH of the mobile phase, applied voltage and gradient elution program. The results indicated that the double separation mechanisms of pCEC could strengthen the separation selectivity and resolution, providing a new analytical platform for the research on Chinese complex prescriptions.3. Setting Berberine and Baicalin as main indexes, the pharmacokinetics of GQD in rat was investigated, by using high performance liquid chromatography (HPLC) method. After intragastric administrating GQD to SD male rats, the concentrations of Berberine and Baicalin in rat plasma were measured at the same time, by methanol-0.5% triethylamine (TEA) isocratic elution, detected at 346 nm and 280 nm, with SHIMADZU LC-20A double-length UV/vis detector. The Concentration-Time curves were drawn. The results showed that Tmax of Berberine and Baicalin were 3 h and 4 h, respectively. The absorption of Baicalin was not regular, multi-peaks appeared.In conclusion, modern chromatography and multiple techniques were utilized for the investigation of separation and analysis of effective ingrediens in Chinese complex prescription GQD. The platforms of high performance liquid chromatography, two-dimensional liquid chroma- tography coupled to mass spectrometry and pressurized capillary electrochromatography were proved to be effective in the analysis of the effective components in GQD, such as flavonoids, alkaloids, and so on. The pharmacokinetics of index components in GQD was also studied. The results of this thesis provided new thoughts and technology platforms for the research on modernization of quality control of the traditional Chinese medicines and their metabolites.