ABSTRACT
OBJECTIVE：To opt imize the extraction technology of phenolic acid from Amomum tsaoko . METHODS ：The extraction technology of phenolic acid from A. tsaoko was optimized by using Box-Behnken design-response surface methodology with ethanol volume fraction ，liquid-solid ratio and extraction time as factors ，using the total contents of protocatechuic acid and vanillic acid as response value. The optimizd extraction technology was vlidated. RESULTS ：The optimal extraction technology was as follows ：ethanol volume fraction 65％，liquid-solid ratio 4∶1（mL/g），extraction time 2.5 h. After 3 times of validation tests ， average total content of protocatechuic acid and vanillic acid were 12.32 mg/g（RSD＝0.26 ％，n＝3），average relative error of which with predicted value （12.63 mg/g）was 2.45％. CONCLUSIONS ：The optimal technology is stable and feasible .
ABSTRACT
OBJECTIVE：To establish a method for the content determination of to tal flavonoids from Amomum tsao-ko ，and to optimize the purification technology by macroporous resin. METHODS ：The content of total flavonoids was measured by HPLC. The determination was performed on Eclipse Plus C 18 column with mobile phase consisted of acetonitrile- 1％ acetic acid solution （15∶85，V/V）at the flow rate of 0.8 mL/min. The column temperature was 40 ℃，and the detection wavelength was set at 256 nm. The sample size was 10 μL. Taking the adsorption and desorption performance as indexes，6 kinds of macroporous resins were screened out by static adsorption and desorption tests ；adsorption and desorption time were investigated by static adsorption and desorption kinetics tests. Using the content of total flavonoids （calculated by rutin ）as index ，with sample concentration ，sample pH，ethanol volume fraction and elution amount as factors ，based on single factor test ，orthogonal design was used to optimize the purification technology of total flavonoids from A. tsao-ko ，and validation test was performed. RESULTS ：The linear range of rutin were 0.028-0.281 mg/mL（r＝0.999 9）. The limit of quantification was 437.5 ng/mL and the limit of detection was 109.4 ng/mL. RSDs of precision ，stability and reproducibility tests were all lower than 2％；the recoveries were 96.24％-99.75％（RSD＜2％，n＝ 6）. The comprehensive capacity of adsorption and desorption of HPD 450 macroporous resin was the most suitable ，and the best static adsorption and desorption time both were 12 h. The optimal purification technology was 1.854 4 mg/mL ； ethanol elution was 8 times of the column volume. Vertificationtests show that after optimized ，the content of total flavonoids from A. tsao-ko increased from 22.556 7 mg/g to 57.728 2 mg/g. The purity of was 2.56 and stable for the content determination. Optimal purification technology is stable and feasible ，which is suitable for purifieation of total flavonoids from A. tsao-ko .
ABSTRACT
OBJECTIVE: To investigate the inhibition effect of total alkaloid, total phenolic acid and total volatile oil from Ligusticum chuanxiong on the contraction of the isolated uterine smooth muscle of rats, and to provide reference for clarifying the promoting blood circulation to restore menstrual flow traditional efficacy of L.chuanxiong. METHODS: The isolated uterine smooth muscle of rats were collected and soaked in Locke's solution. Using dimethyl sulfoxide as blank control, verapamil hydrochloride as positive control, the contraction curves of the isolated uterine smooth muscle in rats were recorded with PowerLab electrophysiolograph. Inhibitory effects of total alkaloid, total phenolic acid (mass concentrations of 0. 025, 0. 05, 0. 1 mg/mL) and total volatile oil (0. 04, 0. 08, 0. 16 mg/mL) on the contraction of the isolated uterine smooth muscle of rats induced by oxytocin were evaluated by observing the changes of contraction activity and contraction tension. Inhibitory rates of contraction activity and contraction tension of uterine smooth muscle were calculated. RESULTS: The contraction activity and contraction tension of uterine smooth muscle in rats were increased significantly after treated with oxytocin (P<0. 05 or P<0. 01). Different concentrations of 3 types of compounds from L. chuanxiong all decreased contraction activity and contraction tension of uterine smooth muscle significantly (P<0. 05 or P<0. 01). Compared with blank control, except for 0. 025 mg/mL total phenolic acid, the inhibitory rates of contraction activity and contraction tension of uterine smooth muscle were all decreased significantly (P<0. 05 or P<0. 01)after treated with other concentration drugs or components. CONCLUSIONS: The total alkaloid, total phenolic acid and total volatile oil from L. chuanxiong show certain inhibitory effect on the contraction of isolated uterine smooth muscle in rats, which provide reference for promoting blood circulation to restore menstrual flow use of L. chuanxiong.