Monitoring In Pharmaceutics Of Sheng-Hua-Tang Decoction By RP-HPLC And Pharmacokinetic Study On Sheng-Hua-Tang Decoction

It is a vital problem for the modernization of Traditional Chinese Medicine (TCM) in the domain of life scientific research now. As a key branch of TCM, pharmacology of Traditional Chinese Medicine Recipe (TCMR) is aimed to study the principle of action for the efficacy of Traditional Chinese Medicine especially of Traditional Chinese Medicine Recipe, and is being a focus of TCM modernized research currently. At present, most of advances of research about pharmacology of TCMR have been obtained, but some research is only to manifest a sort of drug effect, by the contrary, the principle research of action is unsubstantial yet. Guided by the hypothesis of Pharmacokinetics of traditional Chinese syndrome and recipe, recipe-derived components spectrum in serum in this paper, we combine the theory of TCM with that of modern medicine. In the experiment process, the recipes of Sheng-Hua-Tang (SHT) and Guan-Xin-Er-Hao (GXEH) that are strictly combined and do works for therapy were used as examples. After the strict control on the quality of Chinese crude drug and the decocting process, the pharmacokinetic research on SHT decoction was made. Then the chemical components from GXEH decoction compared with that from human serum after the administration of GXEH decoction were discussed preliminarily.Methods: 1. To compare the contents of ferulic acid in SHT decoctiondecocted together and decocted separately, to study the effects of different preparation of Glycyrrhiza uralensis Fisch.and Zingiber officinal Rosc, in SHT decoction on the contents of ferulic acid; and to compare the contents of feruLic acid in SHT decoction decocted in different solvent, RP-HPLC was used to detect the contents of ferulic acid. Using Inertsil ODS-3 C₁8 column (250×4.6mm, 5 μ m), the mobile phase was methanol-water (containing 1% acetic acid) (45:55,v/v) with flow rate of 1 ml min^-1 and at the detection wavelength of 321nm, and the contents were calculated by external-standard method. 2. FA in serum after SHT decoction was orally administered by the 11 healthy female volunteers was determined by RP-HPLC. The serum samples were extracted and deproteinized with boiling water. A Inertsil ODS-3 C₁8 column (250×4.6mm, Sum) 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. And p-hydroxybenzaldehyde was used as the internal standard. The 3P97 program was used to calculate the pharmacokinetic parameters. 3. After GXEH decoction was orally administered to the healthy male volunteers, the serum sample at the time of 30min which will be analyzed for RP-HPLC was extracted and deproteinized with acetonitrile, methanol and boiling water, respectively. Isocratic and gradient elution were performed respectively for each sample. HPLC-fingerprints(HPLC-FPS) of GXEH decoction and serum obtained from the volunteers treated with GXEH decoction were compared with each other.Results: 1. The calibration curve was linear in the range of 0.6 180.0 mg L^-1 (r=0.9999, n=6). The repeatability (RSD) of the method was generally less than 10% (n=5, inter-day and intra-day). The average recovery was up to the mustard. The content of ferulic acid in SHT decoction decocted together was higher than that in SHT decoction decocted separately(P<0.01). Preparation has effect on the contents of ferulic acid (P<0.01). Proper preparation could increase the release of ferulic acid, that is to say, traditional preparation produced greatest contents of ferulic acid. Decocting in differentsolvent have influence on the contents of feruLic acid (P<0.01) and the contents were highest in SHT decoction decocted in soLvent containing half millet wine and half childish urine. 2. FA with the quantitation limit of about 8.0 g L^-11 (S/N=3) was measured by RP-HPLC separation with UV detection at 321 nm. The calibration curve was linear (r=0.9984) in the concentration range of 12.0 to 360.0 g L^-1. Both of the intra- and inter-day precision of FA were determined and their coefficience of variation did not exceed 10%. The average recovery(n=5) was (99.88 ±4.9)%. This method has been successfully applied for pharrnacokinetic studies of FA from human serum after oral administration of SHT decoction. The main pharmacokinetic parameters of FA were as follows: T₁/2 =18.72min, T₁/2 3 =79.2lmin, T₁/2 Ka=11.19min, AUC=18004.87 g min L^-1, V/Fc=7.32L kg^-1, CL=0.17L min^-1 kg^-1, C_max=206.30 ug L^-1 T_peak =22.78 min. 3.The number of chromatographic peaks detected from GXEH decoction was much smaller than that of chemical components found in Chinese crude drug and the chromatographic peaks detected in medicated serum were much less than those of the TCMR decoction. Only FA from human serum after oral administration of GXEH decoction was successfully analyzed qualitatively.Conclusion: 1. The traditional method of decocting herbs is reasonable and proper herbal preparation is scientific. 2. Establishing a determination method of FA from human serum and the method is simple, rapid, sensitive, reproducible and no toxic. That the effective, chemical component (FA) absorbed into circulation after the administration of SHT decoction could be determined qualitatively and quantitatively was proved firstly. FA in healthy female volunteers after oral administration of SHT decoction could be absorbed and eliminated rapidly. The FA pharmacokinetics conforms to a two compartment open model. 3. The chromatographic peaks detected in medicated serum were much less than those of the TCMR decoction .The results suggest that the active components of TCMR in vivo might be "relatively limited".Above all, based on the test of PKT of SHT decoction, the...