Researches On The Determination Of Effective Components Of 4 Compound Traditional Chinese Water-Decocted Medicines And Their Effects On The Isolated Intestines Smooth Muscles In Rats

Objective: This research comprises two parts. The first part aims to establishmeasure methods for four kinds of traditional Chinese compound water-decoctedmedicines including Jin Ling Zi San, Shao Gan Decoction, Tong XieYao Fang, ChaiYu Decoction by using of RP-HPLC method, to guarantee the safety and efficiency ofthe clinic medicines. The quality control index of Jin Ling Zi San isTetrahydropalmatine in Corydalis yanhusuo W.T.Wang. The quality control index ofShao Gan, and Tong XieYao Fang is Paeoniflorin in Paeonia lacti flora Pall. And thequality control index of Chai Yu Decoction is Curcumin in Curcuma yujin Y.H.Chenet C.Ling. The best chromatogram separation and measurement conditions forTetrahydropalmatine, Paeoniflorin, and Curcumin are chosen, which are strictlyreviewed by means of methodology including precision test of apparatus system,repeatable test of samples, stable test of samples, test of recovery rate of sample toensure the test repeatable, stable, and reliable. The second part of this researchfocuses on the taking stock of influence on the movement of Shao Gan Decoction,Tong Xie Yao Fang and Chai Yu Decoction to the isolated smooth muscles by usingisolated smooth muscles test. Pharmic preliminary choose of three traditional Chinesewater-decocted medicines was made and its action mechanisms was discussed. Thepaper comprised five tests.Methods: At first, on the base of chemical component, carry out the quantitativeresearch on the effective component of JinLingZi San., ShaoGan Decoction, TongXie Yao Fang and Chai Yu Decoction.To establish a reserved-phase HPLC method forthe determination of Tetrahydropalmatine in Jin Ling Zi San. , Paeoniflorin in ShaoGan Decoction, Paeoniflorin in Tong Xie Yao Fang ,and Curcumin in Chai YuDecoction. Kromasil C₁₈ column (250 mm×4.6 mm, 5μm) was applied, usingmethanol -0.1%phosphoric acid (adjusted pH to 6.0 with triethy-lamine) (60: 40) asthe mobile phase for the determination of Tetrahydropalmatine, the detectionwavelength was 280 nm and the flow rate was 1.0 ml/min with the column temperature of 30℃. With methanol-water (25: 75) as the mobile phase, thedetection wavelength was 230 nm and the flow rate was 1.0 ml/min with thecolumn temperature of 30℃for the determination of Paeoniflorin. For thedetermination of Curcumin in Chai Yu Decoction with methanol -water (70: 30) asthe mobile phase, the detection wavelength was 430 nm and the flow rate was 1.0ml/min with the column temperature of 30℃. On the second part of study, to appendintestine tube of rats, the index is contraction intension of isolated ratintestines. Observing on the effect of Regulating the Flow of Qi of Chinese Drugs and4 kinds serum on isolated intestinal smooth muscle, and effect of convulsion causingby the vary exciting medicament, making further effort to observe the medicineactivity of the sample. Finally, experiment evidence is provided for the clinic using offour compound medicines by comprehensive analysis of pharmic choosing result,physical basis and activity mechanisms, as well as the relation of the three aspects.Results: Having established a reserved-phase HPLC method for thedetermination of Tetrahydropalmatine in Jin Ling Zi San, Paeoniflorin in TongXie Yao Fang and Shao Gan Decoction, Curcumin in Chai Yu Decoction. The linearresponse range of Tetrahydropalmatine was 10.048～251.2μg/ml (r=0.99998, n=7);The average recovery rate was 102.4%, RSD=0.83%(n=6). The linear response rangeof Paeoniflorin was 14.24～142.4μg/ml (r=0.99993, n=7), the average recovery rateof Paeoniflorin in Shao Gan Decoction was 101.98%, RSD=1.79% (n=6). Theaverage recovery rate of Paeoniflorin in Tong Xie Yao Fang was 100.07%,RSD=1.96% (n=6). There are no the same chromatogram peaks in the relevantpositions as the comparative ones with the same remaining time in the sample testliquid, after many tests of determination of Curcumin in ChaiYu Decoction. TongXieYao Fang ( 10^-4 g/L), Shao Gan Decoction ( 10^-4 g/L) all inhibited thespontaneous contraction significantly (P＜0.01), decreased the mean contractilefrequency and declined the swing of the contract and was dose-dependent.The effectwas antagonized by Ach, BaCl₂, CaCl₂ and could in crease the inhibitory action. Chai Yu Decoction and 4 kinds serum had not evidence influence on contractile of isolatedintestinal smooth muscle. Tetrahydropalmatine, Paeoniflorin and Curcumin allinhibited the spontaneous contraction significantly.Conclusion: 1. The method is sensitive and accurate with good reproducibility. It issuitable for the studying on the content of Tetrahydropalmatine in Jin Ling Zi San,Paeoniflorin in Tong XieYao Fang and Shao Gan Decoction. 2. Tong XieYao Fangand Shao Gan Decoction and monocase of Paeoniflorin, Tetrahydropalmatine andCurcumin have had distinct inhibitory action on the movement of isolated intestinalsmooth muscle of rats, and dose-depended, which may be mediated through Achreceptor, Ca²⁺ channel or directly act on the intestinal muscle.