| Background: One of the primary obstacle to the modernization of traditional Chinese medicine is that the material base for the efficacy of Traditional Chinese Medicine Recipe(TCMR) has not been clear yet. An important way to clarify the material base for the efficacy of TCMR is to research on the chemical components absorbed into circulation after the administration of TCMR. Experiments indicated that the chemical components absorbed into circulation after the administration of TCMR can be determined qualitatively and quantitatively. The inherent relationship which may exist between the chemical components absorbed into circulation after the administration of TCMR and the efficacy of TCMR has not been elucidated scientifically yet, which is the key problem at present as well as the difficulty of the research works on the material base for the efficacy of TCMR. To solve this problem, quantitative and systematic studies can be introduced to study the relationship between pharmacokinetics(PK) of the chemical components absorbed into circulation after TCMR were administered and the pharmacodynamics(PD) of TCMR in vivo. Nowadays, study on PK-PD integration of TCMR is little. And there is no report on clinical PK-PD integration studies on TCMR.Aim: In our experiments, the recipe of Guan-Xin-II (GXEH) that is strictly combined and does works for therapy was used as an example. Thesimultaneous determination method of the multiple components in TCMR and the observing techniques on coronary hemodynamics by noninvasive echocardiography were established after the strict control on the quality of Chinese crude drug and the decocting process. Then the clinical PK.-PD integration studies of GXEH were performed.Methods: 1. A ZORBAX Extend-Ci8 reversed-phase column (particle size 5 u m, 150X4.6 mm) was used as the stationary phase. The mobile phase was composed of A (MeOH-HAC 99: 1, v/v)-B (Water-HAC 99: 1, v/v) with linear gradient elution (0 min, 15: 85; 25 min, 40: 60). The flow rate was 1 ml/min. The detection of Danshensu (Dhpl), protocatechuic aldehyde (Pal), paeoniflorin(Pa) and ferulic acid(FA) were performed at wavelength of 280 nm,230 nm and 321 nm , respectively at room temperature. A reverse phase high performance liquid chromatographic(RP-HPLC) method for simultaneous determination of Danshensu (Dhpl), protocatechuic aldehyde (Pal), paeoniflorin(Pa) and ferulic acid(FA) in GXEH decoction was established. 2.The strict quality control of the Chinese crude drug including the growing area, the channel to be obtained and authentication of the drugs as well as the manual decocting process including the amount of decocting water, soak duration and decocting duration and firepower of soft fire and hot fire were conducted. 3. After GXEH decoction was orally administered to the normal male volunteers, the serum sample at the time of 30min that will be analyzed for HPLC was extracted and deproteinized with acetonitrile, methanol and boiling water respectively. GXEH decoction was properly diluted for analysis too. Isocratic and gradient elution were performed respectively for each sample with a diode-array UV/V with multi-wavelength detector scanning between 200 to 400nm. HPLC-fingerprints(HPLC-FPS) of GXEH decoction and serum obtained from the volunteers treated with GXEH decoction were compared with each other to detect serum chemical components from GXEH decoction. 4. FA in serum at the time of 10,20,30,45,60,90,120, 180min after the GXEH decoction was orally administered by the normal male volunteers was determined by HPLC. The serum samples were extracted anddeproteinized with boiling water. A Inertsil ODS-3 C18 reversed-phase column (particle size 5 u m, 250 X4.6mm) was used as the stationary phase. The mobile phase was methanol-0.6% acetic acid (43:57,v/v) with flow rate of 1 ml/min. Detection was performed at a wavelength of 321nm. p-hydroxybenzaldehyde was used as the internal standard. The pharmacokinetical parameters were fitted with software 3P97. 5. Following the validating Color Doppler Coronary Artery Flow...
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