Studies On Effective Substances Of Huoxiang Zhengqi Liquid On Intestinal Spasm And Its Quality Control

Huoxiang Zhengqi Liquid, which was recorded originally in Tai Ping Hui Ming He JiJu Fang and was a famous Chinese herbal formula in China, was studied by the optimizingstrategy gradually and hierarchically. Its the effective substances having intestinal spasmwere researched gradually through the levels of chemical groups and chemicals. Weestablished its quality control method by adopting suitable analysis means. Main researchcontents were included as follows.1. Using the inhibition ratio of rat duodenum in vitro as the pharmacological indexeseffected by the intestinal contents of Huoxiang Zhengqi Liquid and the best time is at25min．The correlation analysis between the datas of the inhibition ratio and the chemicaldatas by SPSS indicated the effective component of the intestinal spasmolysis in theHuoxiang Zhengqi Liquid．2. Based on the correlation analysis of effective component group, silica gel columnchromatography, preparative thin layer chromatography, and recrystallization were used toisolate the compounds from the chloroform extract．The structures of the compounds wereidentifid by spectral analysis and physicochemical properties．Fifteen compounds wereobtained and identified as liquiritin（1）、liquiritigenin（2）、isoliquiritigenin（3）、formononetin（4）、 oxypeucedanin hydrate（5）、 byakangelicin（6）、 hesperidin（7）、5,7,8,3’,4’-pentamethoxyflavone（8）、5,6,7,3’,4’-pentamethoxyflavone（9）、5,7,8,4’-tetramethoxyflavone（10）、nobiletin（11）、3,5,6,7,8,3’,4’-heptamethoxyflavone（12）、tangeretin（13）、honokiol（14）、magnolol（15）．All the compounds were isolated fromHuoxiang Zhengqi Liquid and the study on their activities was carried out． 3. To study the effects of nobiletin, tangertin, liquiriti, liquiritigenin, isoliquiritigenin,honokiol, magnolol and hesperidin on the contractility in ileum smooth muscle in vitro．Thesamples of isolated ileum segment were prepared and the effects of nobiletin, tangertin,hesperidin, liquiritin, liquiritigenin, isoliquiritigenin, honokiol and magnolol at variousconcentration on the spontaneous contraction of isolated intestine induced by KCl,acetylcholine or histamine phosphate were studied．There were a certain dose effectrelationship between nobiletin, tangertin, liquiritin, liquiritigenin, isoliquiritigenin, honokioland magnolol. The study has explored the spasmolytic effect of Huoxiang Zhengqi Liquidon in vitro intestinal muscle.4. To develop a HPLC method to determine the contents of liquiritin, hesperidin,oxypeucedanin hydrate and byakangelicin in Huoxiang Zhengqi Liquid．It was eluted ingradient mode using Diamonsil C₁₈column．The detection wavelength was set at276nmand the flow rate was1.0mL·min^-1．A good linearity of four components were found overthe concentration range of0.00866～0.17325μg,0.0734～1.46793μg,0.01384～0.13838μgand0.00528～0.10553μg, respectively．The average recoveries were100.7%,99.1%,98.5%and100.4%respectively. This method is simple, accurate, and specific and can beused as a complementary method for the determination of liquiritin, hesperidin,oxypeucedanin hydrate and byakangelicin in Huoxiang Zhengqi Liquid．5. To establish a quantitative method to determine the contents of nobiletin,5,7,8,4’-tetramethoxyflavone, tangertin, imperatorin, honokiol, isoimperatorin, and magnolol inHuoxiang Zhengqi Liquid by HPLC．It was eluted in gradient mode using Gracesmart C₁₈column．The detection wavelength was set at280nm and the flow rate was1.0mL·min^-1. Agood linearity of seven components were found over the concentration range of0.00238～0.11875μg,0.00910～1.45664μg,0.00353～0.77550μg,0.01770～0.35400μg,0.28504～4.47920μg,0.00516～0.24080μg and0.23560～4.71200μg, respectively．The averagerecoveries were100.5%,99.7%,100.7%,101.0%,98.8%,101.8%, and101.3%,respectively. According to effective substances having intestinal spasm, the determinationmethod of multi components in Huoxiang Zhengqi Liquid have been established.