百令胶囊中17种氨基酸的柱前衍生化RP-HPLC法含量测定  

Pre-column derivatization RP-HPLC determination of 17 amino acids in Bailing capsules

作　　者：邹秦文[1] 肖新月[2] 程显隆[2] 刘燕[2] 张昀[3] 林瑞超[2] 

机构地区：[1]北京中医药大学,北京100102 [2]中国药品生物制品检定所,北京100050 [3]杭州中美华东制药有限公司,杭州310011

出　　处：《药物分析杂志》2010年第9期1630-1635,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：国家科技重大专项课题“中药质量标准研究和信息化体系建设平台”(2009ZX09308-004)专题内容

摘　　要：目的:测定与分析百令胶囊中17种氨基酸的含量。方法:样品以6mol·L-1盐酸于150℃水解1h,以异硫氰酸苯酯(PITC)为衍生化试剂衍生后进行HPLC分析。采用Phenomenex GeminiC18(250mm×4.6mm,5μm)色谱柱,柱温为43℃;流动相A为0.1mol·L-1醋酸-醋酸钠缓冲液(pH6.5)-乙腈(93∶7),流动相B为乙腈-水(4∶1),梯度洗脱,流速为1.0mL.min-1;检测波长为254nm。结果:17种氨基酸浓度在1.02～43.32μg·mL-1范围内呈良好的线性关系(r=0.9996～0.9997);平均回收率除缬氨酸、甲硫氨酸为89.3%和84.8%外,其余15种氨基酸在94.5%～106.7%之间;RSD除苏氨酸、缬氨酸、甲硫氨酸为3.2%,3.6%,4.6%外,其余均小于3.0%。结论:本方法灵敏、准确,具有良好的重复性和稳定性,可用于百令胶囊中氨基酸的检测。Objective：To determine and analyze 17 amino acids in Bailing capsules.Methods：The sample was hydrolyzed for 1 hour at 150 ℃,and derived with phenyl isothiocyanate（PITC）.HPLC was performed on a Phenomenex C18（250 mm×4.6 mm,5 μm）column with gradient elution of 0.1 mol·L-1 acetic acid-sodium acetate buffer solution（pH 6.5）-acetonitrile（93：7）（A）and acetonitrile-water（4：1）（B）at the flow rate of 1.0 mL·min-1,detected at 254 nm.Results：17 amino acids had good linearity in the ranges of 1.02-43.32 μg·mL-1（r=0.9996-0.9997）.The average recoveries were 94.5%-106.7%,except Val 89.3%,Met 84.8%,and RSD were less than 3.0%,except Thr 3.2%,Val 3.6% and Met 4.6%.Conclusion：The method is sensitive and accurate with high repeatability and stability,which is helpful for the determination of amino acids in Bailing capsules.

关 键 词：百令胶囊 衍生化 氨基酸 异硫氰酸苯酯 高效液相色谱 

分 类 号：R917[医药卫生—药物分析学]