RP-HPLC柱前衍生化测定熊去氧胆酸的含量

Determination of ursodeoxycholic acid by pre-column derivation on HPLC

采用反相高效液相色谱柱前衍生化法测定熊去氧胆酸的含量.衍生化条件:以猪去氧胆酸为内标,溴苯乙酮为衍生化试剂,三乙胺为催化剂,8o℃温度下反应30min.色谱条件:色谱柱为Symmertu-C18柱(250*4.6mm,10μm),流动相为甲醇-水(85:15),检测波长为254nm.结果熊去氧胆酸线性范围为0.02～10mmol·L-1,r=0.9998,重复性试验RSD=2.91%(n=6).所建立的反相高效液相色谱柱前衍生化法能准确测定熊去氧胆酸的含量,重复性好.

To develop the determination method of ursodeoxycholic acid by a pre-column derivation. An HPLC method is used with Hypersil C18 column （250 mm×4.6 mm,10 μm） and UV detection at 254 nm. The mobile phase consists of methnol-water（85：15）. Sample is derivatized with phenacyl bromide at 80℃ water bath for 30 minutes,using triethylamine as the catalyst. The calibration curve is linear（r = 0.9998） within the range of 0.02 - 10 mmol·L^-1 for ursodeoxycholic acid, the repeatability is 2.91% （RSD）. The method has a good accuracy and repeatability.