柱前衍生反相高效液相色谱法测定血清中9种胆汁酸  

Simultaneous Determination of 9 Bile Acids by RP-HPLC with Pre-column Derivatization

作　　者：丁敏[1] 刘春芳[1] 何俊[1] 

机构地区：[1]重庆医科大学医学检验系,重庆400016

出　　处：《分析科学学报》2004年第5期492-494,共3页Journal of Analytical Science

摘　　要：建立了一种柱前衍生反相高效液相色谱(RP HPLC)测定血清中9种胆汁酸的方法。以4 溴甲基 7 甲氧基香豆素(BMC)为衍生剂,用WatersNovaC18色谱柱分离,采用甲醇/乙腈和水梯度洗脱,荧光检测,9种胆汁酸在5～40μmol/L范围内具有良好线性,r为0.9988～0.9998。回收率为85.40%～100.45%,检出限为1～2μmol/L。该法灵敏、特效、重复性好,可用于临床血清中微量胆汁酸的测定。A method for the determination of bile acids in serum by RP-HPLC was established. Quantification was obtained using lauric acid (LA) as an internal standard. Nonconjugated and glycine-conjugated bile acids were derivatized with 4-bromomethyl-7-methoxycoumarin in the presence of dicyclohexano-18-crown-6 as a catalyst. Derivatized bile acids were then eluted using an acetonitrile/methanol/water gradient on a Waters Nova C18 column(3.9×150 mm,5 μm). The column was kept at 25℃. The eluate was monitored by a fluorophotometer at 330 nm(λ_(ex)) and 410 nm(λ_(em)). The assay was linear in the range 5～40 μmol/L. Correlation coefficients for linear regression ranged from 0.9988 to (0.9998.) Recoveries from serum was greater than 85%. Precision was within 8%. The analysis time was 24 min. It has been proved that the proposed method was sufficiently sensitive and reliable for the routine analysis of bile acids in serum.

关 键 词：RP-HPLC 胆汁酸 荧光检测柱前衍生化 

分 类 号：O657.72[理学—分析化学] O657.39[理学—化学]