[关键词]
[摘要]
目的 建立高效液相色谱-串联质谱法(HPLC-MS/MS)测定2型糖尿病患者血浆中瑞格列奈的浓度。方法 采用Zorbax Eclipse XDB-C18色谱柱(150 mm×4.6 mm,5μm);流动相:5 mmol醋酸铵水溶液(含0.1%甲酸)-乙腈(10:90);柱温:35℃;体积流量:0.5 mL/min;进样器温度为常温;进样体积:5 μL,进样时间:6 min。采用电喷雾离子源(ESI),正离子多反应离子监测(MRM)扫描,瑞格列奈、内标那格列奈的检测离子对分别为:m/z 453.6→230.5、m/z 318.4→166.3。采集规律服用瑞格列奈片的2型糖尿病患者静脉血约4 mL,采用HPLC-MS/MS法测定患者血药浓度。结果 瑞格列奈在0.1~50 ng/mL线性关系良好,定量下限为0.1 ng/mL。在定量下限(0.1 ng/mL)和低、中、高质量浓度下,准确度为91.71%~103.67%,日内、日间精密度的RSD均小于10%。在低、高两种质量浓度下,瑞格列奈基质效应和提取回收率分别为96.97%~97.62%和81.22%~93.94%,且稳定性较好。结论 本方法简单、经济、快速、灵敏,可用于2型糖尿病患者人体血浆中瑞格列奈的测定。
[Key word]
[Abstract]
Objective To establish a method HPLC-MS/MS for the determination of repaglinide in plasma of patients with type 2 diabetes mellitus. Methods The separation was carried out on Zorbax Eclipse XDB-C18 chromatography column (150 mm×4.6 mm, 5 μm). The mobile phase consisted of 5 mmol ammonium acetate aqueous solution (0.1% formic acid) and acetonitrile (10:90). Temperature of column was set at 35℃, and the flow rate was 0.5 mL/min. Sampler temperature was set at room temperature. The injection volume was 5 μL, and the injection time was 6 min. Electrospray ion source and positive ion multiple reactions monitoring scanning were used. The ion pairs of repaglinide and internal standard nateglinide were m/z 453.6→230.5 and m/z 318.4→166.3, respectively. The venous blood of type 2 diabetes mellitus patients who regularly po administered with Repaglinide Tablets was collected for 4 mL. The blood concentration was determined by HPLC-MS/MS. Results The linear ranges of repaglinide were 0.1-50 ng/mL with the LLOQ of 0.1 ng/mL. At the LLOQ (0.1 ng/mL), and the low, medium and high mass concentrations, the accuracy were 91.71% -103.67% with RSD of inter-day and intra-day precisions less than 10%. At low and high concentrations, the matrix effect and extraction recovery were 96.97%-97.62% and 81.22%-93.94%, respectively, with good stability. Conclusion The method is simple, economical, rapid and sensitive, can be used for the determination of reaglinide in human plasma of patients with type 2 diabetes mellitus.
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[基金项目]
国家自然科学基金面上项目(81173134);皖南医学院中青年科研基金项目(WK2016F06)