| The study of pharmacodynamics components in Chinese medicinal herbs is one of the key issues of modernization of Traditional Chinese Medicine (TCM). In this study, we used modern analytical techniques, from in vitro to in vivo, established a series of research method for the study of pharmacodynamics components. Determination of multi-component can strictly control the content of key ingredients in TCM, and the fingerprint's compounds group represents the chemical composition of Chinese medicine. Utilizing the modern analysis technologies HPLC-TOF/MS, HPLC-MS etc. to identify chromatography peaks in fingerprint, and further identify relevant components in rat plasma and urine, helping us to screen pharmacodynamics components and do mechanism study. Pharmacokinetics study of traditional Chinese medicine get the dynamic change process of major components in the body, and provided perspective for the pharmacodynamics study of TCM, and provide a good guide for the mechanism study.HPLC-TOF/MS technology was applied for the identification of chemical composition of Radix Saposhnikoviae (RS). The accurate molecular weight measurement of time of flight mass spectrometry helped us to identify a total of 32 ingredients, including 8 chromone and 14 coumarins. We create an undoubted fingerprint of RS, providing a workable solution to the analysis of chemical ingredients of TCM. Then, RS from different geographical locations were analyzed and classified via HPLC-TOF/MS combing with multivariate statistical analysis technique for the first time. Principle component analysis (PCA) showed that these samples could be roughly separated into three groups, and through the loading plot of partial least squares-discriminant analysis (PLS-DA), we found chemical markers such as 4'-O-glucosyl-5-O-methylvisamminol, cimifugin, prim-o-glucosylcimifugin and 3'-O-angeloylhammaudol which are important for the classification of RS. Results of this study indicated that it was an effective and novel approach to identify traditional Chinese medicine (TCM) from different sources, while performing quantity determination of corresponding marker compounds could optimize the quality control of TCM.Then, a HPLC-TOF/MS method was developed and validated for the identification of the chemical constituents and metabolites of RS in rat plasma and urine.6 constituents of RS were found in plasma,8 constituents and 2 metabolites were identified in urine. The results showed that cimifugin is the major component in rat plasma and urine and may be the effective component of RS. It's important to control its quality as well as prim-o-glucosylcimifugin and 4'-O-glucosyl-5-O-methylvisamminol. This study was useful for the further study of the clinical application of RS.According to the result of identification of RS ingredients in vitro and in vivo, we established a HPLC-DAD method for the quantitative determination of four chromones including prim-O-glucosylcimifugin, cimifugin,4'-O-glucosyl-5-O-methylvisamminol and sec-o-β-D-glucosylhammaudol. And the method was methodology validated and its results showed that it could be used for the determination of the four compounds. Then,10 batches of RS were determined. This method was of high sensitivity, good stability, low cost, versatility, could be used for routine quality control of RS and provided a strong base for further pharmacological studies.To further study the major components' absorption, metabolism process in the body, we established a HPLC-MS method for the quantitative determination of three chromones including prim-O-glucosylcimifugin, cimifugin and 4'-O-glucosyl-5-O-methylvisamminol in rat plasma, and pharmacokinetic study was completed. Three components' linear range were of 12.99～230.5 ng·mL-1,12.81～24350 ng·mL-1,13.42～124.25 ng·mL-1 respectively; The low, middle and high concentration levels' absolute recoveries were all between 94% and 104%; Precision RSD were less than 15%; The results of stability experiment also meet the requirements. Endogenous compounds didn't obstruct the determination of targeted components; this method is simple and reliable for determination of the components in biological samples. This method has been successfully applied to SD rats who orally administered RS extract solution and prim-O-glucosylcimifugin monomer solution, we compared their concentration-time curves, and found that the pharmacokinetic parameters (AUCo-t, AUCo-∞and MRT) of rats who orally administered RS extract were significantly different from who had prim-O-glucosylcimifugin monomer solution (P <0.05). Prim-O-glucosylcimifugin largely transformed to cimifugin when it was absorbed into blood. The concentration-time curves of rats who orally administered RS extract solution had two peaks, while which of rats who orally prim-O-glucosylcimifugin monomer solution only had one peak and the metabolic speed was fast. These results suggest that there exsited transform traction when the compositions were absorbed into blood and there maybe exsited enterohepatic circulation. To control the quality of RS, it is necessary to determine prim-O-glucosylcimifugin, cimifugin and 4'-O-glucosyl-5-O-methylvisamminol at the same time. This study guides the clinical medication and further pharmacological studies.
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