建立HPLC法同时测定复方黑骨藤中9种有效成分的含量  

Simultaneous determination of nine effective ingredients in compound Heiguteng by HPLC

作　　者：徐艳[1] 李琪[1,2] 姚瑾[1] 李慧敏[1] 张晓峰[1] 张宏[1,2] 

机构地区：[1]四川师范大学生命科学学院,成都610101 [2]四川师范大学植物资源应用与开发研究所,成都610101

出　　处：《药物分析杂志》2016年第7期1190-1198,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金资助项目(编号:31240087)

摘　　要：目的:建立HPLC法同时测定复方黑骨藤中落干酸、6’-O-β-D-葡萄糖基龙胆苦苷、獐芽菜苦苷、龙胆苦苷、獐芽菜苷、杠柳苷、延胡索乙素、四氢小檗碱、延胡索甲素9种有效成分的含量。方法:采用Kromasil C18色谱柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.05%三乙胺+乙酸水溶液(B,pH=6.0)为流动相,梯度洗脱(0~5 min,5%A→8%A;5~5.5 min,8%A→11%A;5.5~20 min,11%A;20~30 min,11%A→30%A;30~40 min,30%A→40%A;40~42 min,40%A→55%A;42~53 min,55%A→75%A),流速1.2 m L·min^(-1),柱温25℃,检测波长为206 nm(延胡索乙素、四氢小檗碱、延胡索甲素)、225 nm(落干酸和杠柳苷)、243 nm(獐牙菜苦苷和獐牙菜苷)和275 nm(6’-O-β-D-葡萄糖基龙胆苦苷和龙胆苦苷),进样量10μL。结果:落干酸、6’-O-β-D-葡萄糖基龙胆苦苷、獐芽菜苦苷、龙胆苦苷、獐芽菜苷、杠柳苷、延胡索乙素、四氢小檗碱和延胡索甲素的质量浓度在一定范围内,与峰面积呈良好的线性关系;平均回收率(n=6)分别为92.7%、90.9%、91.2%、94.9%、97.4%、94.6%、96.1%、90.9%和104.4%,RSD分别为2.1%、1.3%、1.1%、1.6%、0.7%、0.6%、1.8%、2.0%和1.4%;样品中上述9种成分含量范围分别为3.464~3.535、1.133~1.178、0.373~0.382、13.598~14.142、0.109~0.112、0.499~0.510、0.114~0.119、0.027~0.028和0.163~0.174 mg·g^(-1)。结论:经方法学验证,本方法可用于黑骨藤复方药物中9种有效成分的分析和含量测定,为复方黑骨藤药物的质量控制奠定基础。Objective： To establish an HPLC analysis method for simultaneous determination of logannic acid,6＇-O-β-D-glucosylgentiopicroside,swertiamarin,gentiopicroside,sweroside,glucoperiplocymarin,tetrahydropalmatine, tetrahydroberberine, and corydaline in compound Heiguteng.Methods： The HPLC analysis was performed on a Kromasil C18 column（250 mm×4.6 mm, 5 μm） with the mobile phase consisted of acetonitrile（A）-0.05% trimethylamine and glacial acetic acid（B,pH=6.0） in gradient mode（0-5 min,5%A → 8%A;5-5.5 min,8%A → 11%A;5.5-20 min,11%A;20-30 min,11%A → 30%A;30-40 min,30%A → 40%A;40-42 min,40%A → 55%A;42-53 min,55%A → 75%A） at the flow rate of 1.2 m L· min-（-1）.The column temperature was 25 ℃ and the detection wavelengths were 206 nm（tetrahydropalmatine, tetrahydroberberine and corydaline）, 225 nm（logannic acid, glucoperiplocymarin）, 243 nm（swertiamarin, sweroside） and 275 nm（6＇-O-β-D-glucosylgentiopicroside, gentiopicroside）.Results： In a certain range, the mass concentrations of logannic acid, 6＇-O-β-D-glucosylgentiopicroside, swertiamarin, gentiopicroside, sweroside, glucoperiplocymarin, tetrahydropalmatine, tetrahydroberberine, corydaline and the peak areas had a good linear relationship.The average recoveries（n=6） of nine components were 92.7% with RSD of 2.1%, 90.9% with RSD of 1.3%, 91.2% with RSD of 1.1%, 94.9% with RSD of 1.6%, 97.4% with RSD of 0.7%, 94.6% with RSD of 0.6%, 96.1% with RSD of 1.8%, 90.9% with RSD of 2.0%, and 104.4% with RSD of 1.4%, respectively.The content ranges of above ingredients in compound Heiguteng were 3.464-3.535 mg·g-（-1） for logannic acid, 1.133-（-1）.178 mg·g-（-1） for 6＇-O-β-D-glucosylgentiopicroside, 0.373-0.382 mg·g-（-1） for swertiamarin, 13.598-（-1）4.142 mg·g-（-1） for gentiopicroside, 0.109-0.112 mg·g-（-1） for sweroside, 0.499-0.510 mg·g-（-1） for glucoperiplocymarin, 0.114-0.119 mg·g-（-1） for tetrahydropalmatine, 0.027-0.028 mg·g-（-1） for tetrahydroberberine and

关 键 词：复方黑骨藤 黑龙骨 秦艽 元胡 落干酸 6’-O-β-D-葡萄糖基龙胆苦苷 獐牙菜苦苷 龙胆苦苷 獐牙菜苷 杠柳苷 延胡索乙素 四氢小檗碱 延胡索甲素 HPLC 

分 类 号：R917[医药卫生—药物分析学]