HPLC-RIF同时测定人血浆中罗格列酮和吉非罗齐

 目的建立反相高效液相色谱-荧光检测(RP-HPLC-RIF)同时测定血浆中罗格列酮和吉非罗齐浓度的方法,用于罗格列酮与吉非罗齐相互作用的研究。方法采用Macherey-NagelNucleodurC₁₈(4.6mm×250mm,5μm)色谱柱,柱温45℃。以乙腈-30mmol·L^-1醋酸铵(内含0.1%甲酸)为流动相,采用梯度洗脱程序。用时间程序调节荧光检测条件,0.10-7.70min时λkex和λem分别为250和370nm,7.70～10.40min时,λex和λem分别为265和365nm,10.40～16.00min时,λex和λem分别为242和300nm。以α-细辛脑为内标,血浆样品用乙腈沉淀蛋白后进样。结果色谱图中罗格列酮、吉非罗齐及α-细辛脑分离良好,血浆中内源性杂质无干扰。罗格列酮和吉非罗齐分别在(5.0～751.3)ng·mL^-1和(0.5～75.4)μg·mL^-1浓度范围内线性良好。最低检测限分别为2.0ng·mL^-1和0.050μg·mL^-1(S/N=3)。罗格列酮和吉非罗齐的日内、日间RSD均小于10.0%,低、中、高3个浓度的方法回收率分别为95.3%～102.8%和90.9%～104.2%。结论本方法结果准确,操作简单,适合于罗格列酮与吉非罗齐合用时药动学研究,也可用于两个药物的单独测定。

Simultaneous determination of rosiglitazone and gemfibrozil in human plasma by RP-HPLC with fluorescence detector

OBJECTIVE To establish a RP-HPLC method for determining rosiglitazone and gemfibrozil.METHODS The separation of rosiglitazone and gemfibrozil was performed on a MACHEREY-NAGEL NUCLEODUR C₁₈ column with gradient elution. The mobile phase consisted of acetonitrile and 30 mmol·L^-1 ammoninm acetate solution ( with 0.1% formic acid). Time program was applied to control fluorescence detector.α-Asari was used as internal standard. The blood sample was pretreated by acetonitrile to precipitate protein before sample injection. RESULTS The compounds were separated well. The calibration curves of rosiglitazone and gemfibrozil in the range of (5.0～751.3) μg·mL^-1 and (0.5～75.4) μg·mL^-1 respectively were linear. The lowest detectable concentrations of rosiglitazone and gemfibrozil were 2.0 ng·mL^-1 and 0.050 μg·mL^-1. The RSDs of inter-day and intra-day were less than 10.0%.The relative recoveries were 95.3%～102.8% and 90.9%～104.2% .CONCLUSION The method is simple, accurate and sensitive. It is suitable to be applied in the pharmacokinetics and interaction study of rosiglitazone and gemfibrozil.