| Kanlisha is a kind of external thermotherapy application in traditional Chinese civil, it is an empirical formula, from western elaboration factory of Beijing in China. It is composed of four Chinese traditional herbs: Radix Angelicae Sinensis, Rhizoma Chuanxiong, Radix Saposhnikoviae and Vicia amoena.Using water, vinegar and ethanol of different concentrations as the extraction solvent, the best extraction solvent was 80% ethanol by studying extractive yield and the contents of cimicifugoside, chuanxiongzine and ferulic acid. The optimal alcohol extraction process of kanlisha was studied by orthogonal design with extractive yield and the contents of cimicifugoside, chuanxiongzine, ferulic acid and quercetin from kanlisha as quality assessment indices. Three factors were studied in this experiment, including the ethanol consumption, duration of extration and times of extration. The results showed that the optimal extraction condition was 80% ethanol consumpted 8 times the amount of material, refluxing for twice and 1.5 hour per time.RP-HPLC methods were established to simultaneously determine cimicifugoside in Radix Saposhnikoviae, chuanxiongzine in Rhizoma Chuanxiong, Ferulic acid in Radix Angelicae Sinensis. The separation of three ingredients was achieved on a Diamonsil C18 column (250 mm×4.6 mm, 5μm) using solvents of acetonitrile and 0.4%(v/v) acetic acid aqueous (to adjust pH 3.5 with triethylamine)(15:85, v/v) as mobile phase. They were detected at 280 nm with flow rate as 1.0 mL·min-1, the column temperature was 25℃and sample size was 20μL. The results indicated that the calibration curve was linear in the range of 14.35～328.0μg·mL-1for ferulic acid(r=0.999 5), 6.91～158.0μg·mL-1 for chuanxiongzine (r=0.999 5), 8.44～135.1μg·mL-1 for cimicifugoside(r=0.999 4), Average recoveries were 99.24%, 99.21%, 99.86% and RSD were 1.2%, 1.5%, 1.3% (n=9). The method is simple and accurate. It can provide a method of quality assessment for kanlisha.RP-HPLC methods were established to determine quercetin in Vicia amoena. Chromatagraphy was conducted on a Diamonsil C18 column (250 mm×4.6 mm, 5μm) at 25℃and 370nm. The mobile phase was acetonitrile-aqueous( containing 0.3% phosphoric acid) (33:67,v/v) at a flow rate 1.0 mL·min-1. The results indicated that the calibration curve was linear in the range of 6.312～101.0 mg·L-1( r=0.999 5) ; The average recovery was 95.1% and RSD was 3.3% (n=9). The method is simple and accurate. It can provide a method of quality assessment for Kanlisha or it's new preparation.Four Chinese traditional herbs of kanlisha were indentified by TLC with the dots clear and reproducible.To establish atomic absorption spectrometry (AAS) determined six inorganic elements and four harmful heavy metals in four Chinese traditional herbs of kanlisha. Linear correlation, precision and recovery of ten elements were all correspond to detection request.The assay methods were accurate and with good reproducibility and can provide a quantitative basis for the quality assessment for kanlisha. |
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