UPLC与HPLC测定人血浆中多索茶碱的比较

Comparison of UPLC and HPLC methods for the determination of doxofylline in human plasma

目的比较UPLC与HPLC分析多索茶碱血药浓度时灵敏度的差异。方法 UPLC法的色谱柱为Kinetex C_(18)柱(100mm×2.1 mm,1.7μm),HPLC法的为Diamonsil C_(18)柱(150 mm×4.6 mm,5μm),流动相均为甲醇-10 mmol·L^(-1)乙酸铵(25∶75),流速分别为0.3、1.0 mL·min^(-1),检测波长均为274 nm,进样量分别为10、20μL。结果与HPLC比较,UPLC法中多索茶碱的灵敏度提高了300倍,同时分析时间缩短1/3;UPLC法能够定量测定临床接受多索茶碱治疗40名患者的血浆药物浓度为0.163~12.8μg·mL^(-1),而其中8名患者的血药浓度低于1.0μg·mL^(-1),HPLC法无法准确测定。结论与HPLC比较,UPLC不仅能缩短分析时间,而且能大幅度提高分析方法的检测灵敏度,可满足临床治疗药物监测对分析方法的灵敏度需求。

OBJECTIVE To compare the differences in sensitivity of the analysis of plasma doxofylline concentration between UPLC and HPLC. METHODS The separation in UPLC was performed on a Kinetex C1s column（ 100 mm x 2.1 mm, 1.7 μm） and the mobile phase consisted of methanol- 10 mmol@L-1 ammonium acetate（25 ： 75） with a flow rate of 0. 3 mL·min-1 and injection volume of 10 μL. A Diamonsil Ctseolumn（ 150 mm x 4.6 mm ,5 μm） was utilized with the same mobile phase as the HPLC method ,with a flow rate of 1.0 mL. min - l and injection volume of 20 μL. The detective wavelength was set at 274 nm for both methods. RESULTS Compared with HPLC ,the sensitivity of UPLC method in doxofylline was increased by 300 times,at the same time, analysis time was shortened 1/3. The quantitative determination by UPLC of plasma doxofylline concentration of 40 patients of 0. 163 - 12. 8 μg· mL- 1 was carried out, and as the blood concentrations of 8 patients were below 1.0 μg · mL-1. However, the established HPLC method could not be accurately measured. CONCLUSION Compared with HPLC, UPLC can not only shorten the analysis time,but also greatly improve the detection sensitivity, and meets the needs of the sensitivity in clinical drug monitoring.