HPLC法测定L-氢化乳清酸中异构体的含量

Determination of Isomers in L-Dihydroorotic Acid by HPLC

建立了高效液相色谱法测定L-氢化乳清酸中异构体的含量.色谱柱为CHIRALPAK QDAX(4.6mm×150mm,5μm),以甲醇—乙腈—乙酸—三乙胺(体积比50∶50∶2∶0.2)为流动相,检测波长为245nm,流速为0.5mL/min,柱温为25℃.该方法经过专属性、检测限、重复性、耐用性和待测溶液稳定性的验证,测定结果表明,L-氢化乳清酸及异构体在此条件下能够良好分离,分离度为3.0;异构体检测限为质量浓度0.7μg/mL;待测溶液室温放置24h内稳定.该方法可成功用于L-氢化乳清酸及异构体的分离,且异构体的含量测定结果准确、可靠.

An HPLC analytical method of isomer in L-dihydroorotic acid was established.The HPLC method was performed on CHIRALPAK QD-AX column（4.6 mm×150 mm,5μm）using methanol-acetonitrile-acetic acid-triethylamine（50∶50∶2∶0.2）as the mobile phase at a flow rate of 0.5 mL/min.The detection wavelength was at 245 nm.The column temperature was 25 ℃.The method was validated for specificity,limit of detection,repeatability,robustness and stability of the solution.The results showed that the resolution between L-dihydroorotic acid and isomer was about 3.0.The limit of detection of isomer was found to be 0.7μg/mL.The sample solution should be stable at room temperature for at least 24 h.The proposed method can be successfully used for separation of isomer fromL-dihydroorotic acid,and the result is proved to be accurate and reliable.