【作者】 郭允；

【导师】 万绍晖； 赵春杰；

【作者基本信息】 河南大学 ， 药剂学， 2008， 硕士

【摘要】 坎离砂为经验方,来自北京市西域加工厂,是由当归、川芎、防风、透骨草四味中药材组成的复方中药,本研究对其提取工艺、质量评价方法及四味中药材中微量元素和重金属的含量测定进行了研究,为其进一步研究开发提供参考。以水、醋、不同浓度乙醇为提取溶剂,以浸膏得率、升麻苷、川芎嗪、阿魏酸的含量为考察指标,确定其最佳提取溶剂为80%乙醇。采用正交试验设计,选取浸膏得率、升麻苷、川芎嗪、阿魏酸、槲皮素的含量五个考察指标,对提取工艺中乙醇用量、提取时间和提取次数三个因素进行优化。然后又通过小鼠热板镇痛试验对优化后的工艺和原工艺进行了药效学比较,结果发现优化工艺的小鼠平均痛域值高于原工艺,最后通过综合评价法确定最优工艺为:全方用8倍量的80%乙醇,水浴回流2次,每次1h。建立了同时测定坎离砂中升麻苷、川芎嗪、阿魏酸含量的高效液相色谱分析方法,采用DiamonsilC18色谱柱（250mm×4.6mm,5μm）,以乙腈-体积分数为0.4%的冰醋酸水溶液（三乙胺调pH3.5）（体积比15:85）为流动相,流速为1.0mL·min-1;检测波长280nm,柱温25℃,进样量20μL。实验结果表明:升麻苷在8.44～135.1μg·mL^-1范围内线性关系良好,相关系数为0.9994,平均回收率为99.86%,RSD为1.2%;川芎嗪在6.91～158.0μg·mL^-1范围内线性关系良好,相关系数为0.9995,平均回收率为99.21%,RSD为1.5%;阿魏酸在14.35～328.0μg·mL^-1范围内线性关系良好,相关系数为0.9995,平均回收率为99.24%,RSD为1.2%;该方法准确、重复性好,为全面评价坎离砂质量提供了一个定量依据。建立了坎离砂中槲皮素含量测定的高效液相色谱法。采用DiamonsilC18色谱柱（250mm×4.6mm,5μm）,以乙腈-体积分数为0.3%磷酸水溶液（体积比33:67）为流动相,流速为1.0mL·min-1,检测波长370nm,柱温25℃,进样量10μL。实验结果表明:槲皮素在6.312～101.0μg·mL^-1范围内线性关系良好,相关系数为0.9995;平均回收率为95.1%,RSD为3.3%。该方法简便、准确、重现性好,为坎离砂及其新剂型改进提供了一种质量控制方法。建立了坎离砂中当归、川芎的同时鉴别及防风、透骨草鉴别的薄层色谱法,方法简便、快速,为坎离砂及其制剂的定性分析提供了参考。建立了原子吸收光谱法测定坎离砂四味中药材当归、川芎、防风、透骨草中6种无机元素镁、锌、铁、锰、钙、铜及4种重金属砷、汞、铅、镉的含量。实验结果表明,各元素线性关系、精密度和回收率均满足测定要求,该法操作方便,成本低,可做为坎离砂及中药材中微量元素和重金属的测定方法。本研究建立的定性、定量分析方法能更加全面地控制该方的质量,并且优化后的提取工艺及6种无机元素的定量测定,可为今后剂型的改进提供基础,并为坎离砂的进一步研究开发提供有用的参考价值。更多还原

【Abstract】 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.更多还原