柱前衍生化-HPLC法测定牛磺熊去氧胆酸中牛磺酸的残留量  

Determination of taurine residue in tauroursodeoxycholic acid by pre-column derivatization and HPLC

作　　者：徐广见 徐纪文 罗素芗 XU Guangjian;XU Jiwen;LUO Suxiang(Zhongshan Belling Biotechnology Co.,Ltd.,Zhongshan 528437,Guangdong,China;Guangdong Food and Drug Vocational College,Guangzhou 510640,China)

机构地区：[1]中山百灵生物技术股份有限公司,广东中山528437 [2]广东省食品药品职业技术学校,广州510640

出　　处：《应用技术学报》2024年第3期311-315,共5页Journal of Technology

摘　　要：建立了测定牛磺熊去氧胆酸(TUDCA)原料药中牛磺酸残留量的柱前衍生化高效液相色谱(HPLC)分析方法。样品用芴甲氧羰酰氯(FMOC-Cl)进行柱前衍生,产物用C18反相色谱柱进行HPLC分离,以柠檬酸-柠檬酸钠缓冲液(A)与甲醇-乙腈-柠檬酸-柠檬酸钠缓冲液(B)为流动相梯度洗脱,检测波长264 nm。该方法能使痕量牛磺酸与牛磺熊去氧胆酸及其有关物质达到基线分离,牛磺酸峰面积与浓度在0.31~41μg/mL范围内线性关系较好,相关系数r为0.9995,检出限为0.1μg/mL,加标回收率在98.8%~100.5%之间。已用于实际样品的分析。该方法灵敏度高,重现性好,适用于牛磺熊去氧胆酸中牛磺酸的残留量的测定。A pre-column derivatization high performance liquid chromatography(HPLC)method was established to determine residual content of taurine in taurodeoxycholic acid(TUDCA)drug substance.The samples were treated with pre-column derivatization reagent 9-fluorenylmethyl chloroformate(FMOC-Cl).The derivative reaction products were then separated by HPLC using reverse phase C18 column,and gradient elution with mobile phase citric acid-sodium citrate buffer(A)and methanol-acetonitrile-citric acid-sodium citrate buffer(B).The UV detection wavelength was 264 nm.By the method,trace amount of taurine could be well separated at baseline level from the other related substances in taurodeoxycholic acid sample.The linear correlation coefficient r between peak area and taurine concentration was 0.9995 in the range of 0.31~41μg/mL.The limit of detection was 0.1μg/mL.The recovery was found between 98.8%~100.5%.The method has been applied in real samples analysis and shown sensitive,reproducibly,and suitable for detecting residual content of taurine in taurodeoxycholic acid.

关 键 词：牛磺熊去氧胆酸 牛磺酸 柱前衍生 芴甲氧羰酰氯 

分 类 号：O657.72[理学—分析化学] R917[理学—化学]