| The Zi-Shen pill was reported originally in Secret Record of the Chamber of Orchids in the years of Yuan Dynasty in China and was listed in the Drug Standard of Ministry of Public Health of the People's Republic of China. The pill is prepared by mixing Rhizoma Anemarrhenae, Cortex Phellodendn and Cortex Cinnamomz with honey together. In the clinical practice of traditional Chinese medical science, the pill has been used to treat prostatitis, ptostatomegaly, and infection of the urinary system diseases over a very long period of time and has produced a quite good effect.The compatibility and therapeutic basis of Zi-Shen pill were studied in this paper. The effect of the pill on antiinflammatory and immune function of mice were selected as pharmacological indices. Using water and alcohol of different concentrations as the extracting solvent, the effects on extracting efficiency with different extracting methods were studied. 50% alcohol was selected as the optimal solvent.The quantitative analysis of the compatibility can be accomplished by Step regression analysis, canonical correlation analysis and variance analysis. To ascertain the compatibility of 3 herbs in a composite prescription, solutions of the different combinations of composite herbal medicines of the Zi-Shen pill, combined the crude drugs without, one or two at a time, Anemarrhenae Rhizoma, Phellodendri Cortex, Cinnamomi Cortex, were preparaed; To ascertain the optimal dose, the L9(34) orthogonal array, which permits to arrange 9 combinations, 3 levers for each herb, were adopted. The index by which the therapeutic effect could be assessed was the pharmacologic action assigned. The combination, which produced the largest pharmacologic index, was the best, and the optimal dose was ascertained. That RhizomaAnemarrhenae and Cortex Phellodendri are principal medicines of the pill, whereas Cortex Cinnamomi is corrigent of it has been confirmed by the pharmacologic experiments, We have successfully established a valid approach to confirm the reasonability of the compatibility of the TCM Zi-Shen pill and its optimal dose.Then, a gradient HPLC method for obtaining the chemical fingerprints was established. HPLC analysis of the 15 combinations were carried out. Correlation analysis and regression analysis were applied to interrelate the chemical and the pharmacologic data. The effects of several chemical constituents in the recipe were elucidated, and the therapeutic basis was defined. Among them, mangiferin in Rhizoma Anemarrhenae, berberine hydrochloride in Cortex Phellodendri and cinnamic acid in Cortex Cinnamomi were selected as the quality indices.The optimal alcohol extraction process of Zi-Shen pill was studied by orthogonal design with extractive yield and the content of mangiferin, berberine hydrochloride and cinnamic acid as indices. Three factors were studied in this experiment, including the solvent consumption, time of extraction and times of extraction. The result showed that the optimal extracting condition was 50% alcohol consummated ten times the amount of material, and three times for 60 minutes each time.Sarsasapogenin in Rhizoma Anemarrhenae, berberine hydrochloride in Cortex Phellodendri and cinnamaldehyde in Cortex Cinnamomi were identified by TLC. Mangiferin, berberine hydrochloride and cinnamic acid in Zi-Shen pill were determined by RP-HPLC. The linear range for them were 59.4-593.6 ug mL-1 (r=0.9997), 24.5-245.0 ug mL-1 (r=1.0000) and 1.5-38.4 ug mL-1 (r=0.9990) respectively. The average recoveries of them were 97.3% (RSD=1.9%), 100.8%(RSD=2.8%), 103.1%,(RSD=1.3%) respectively. Gradient elution was employed to determine the contents of five marker substances such as mangiferin, jateorrhizine, palmatine, berberine, cinnamic acid in the pill, five regression equations showed linear relationships between the peak area of each marker and concentrations. The recoveries of the markers listed above were 95.5%(RSD=2.5%), 98.3%(RSD=1.5%), 96.8%(RSD=1.0%), 99.5% (RSD=1.0%), and 101.7%(RSD=2.9%) respectively. Above methods...
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