二维液相色谱检测脑胶质瘤患者同步放化疗及辅助化疗期间的血浆替莫唑胺峰浓度

Peak concentration of plasma temozolomide in patients with glioblastoma during concurrent chemoradiation and adjuvant chemotherapy by two-dimensional liquid chromatography

目的 建立胶质瘤患者血浆替莫唑胺峰浓度的快速测定方法,并分析同步放化疗及辅助化疗期间其血药浓度分布的特点。方法 血浆以盐酸酸化后加冰乙腈去蛋白处理,离心分取上清。样本经Aston SNCB一维色谱柱(4.6 mm×50 mm,5 μm)在线萃取,在Aston SBR二维色谱柱(4.6 mm×250 mm,5 μm)色谱分离。一维流动相为 MVV-1C型样本萃取液,流速为0.8 mL·min^(-1);二维流动相由OPI-2A型、BPI-2C型、API-2C型、MPI-2A型样本萃取液组成,流速为1.2 mL·min^(-1);进样量200 μL,柱温40℃,检测波长330 nm。结果 血浆替莫唑胺在0.4~25.6 μg·mL^(-1)与峰面积线性关系良好(r=0.9995),转移回收率大于96.50%,提取回收率大于95.72%,日内、日间准确度和精密度良好。测定53例患者血浆样本,同步放化疗期间患者替莫唑胺剂量标准化平均峰浓度明显大于其辅助化疗期间(8.25 μg·mL^(-1) vs 5.82 μg·mL^(-1));同步放化疗期间男性剂量标准化平均峰浓度低于女性(5.86 μg·mL^(-1) vs 12.33 μg·mL^(-1))。结论 该方法简便、快速、准确,适合替莫唑胺血浆峰浓度监测。

Objective To rapidly determine the peak concentration of temozolomide in the plasma of patients with glioblastoma,and to analyze the distribution characteristics of its plasma concentration during concurrent chemoradiation and adjuvant chemotherapy.Methods The plasma was acidified with hydrochloric acid and deproteinized with glacial acetonitrile before centrifugation to extract the supernatant.The sample was extracted online by an Aston SNCB one-dimensional chromatographic column(4.6 mm×50 mm,5μm).The chromatographic separation was done by an Aston SBR twodimensional chromatographic column(4.6 mm×250 mm,5μm).The one-dimensional mobile phase was MVV-1C type sample extraction liquid,at a flow rate of 0.8 mL·min^(-1).The two-dimensional mobile phase was composed of OPI-2A,BPI-2C,API-2C,and MPI-2A sample extractions,with a flow rate of 1.2 mL·min^(-1).The injection volume was 200μL.The column temperature was 40℃,and the detection wavelength was 330 nm.Results Plasma temozolomide showed good linearity(r=0.9995)at 0.4~25.6μg·mL^(-1).The transfer recovery rate was greater than 96.50%,the extraction recovery rate was greater than 95.72%,and the intra-day and inter-day accuracy and precision were all good.The concentrations of temozolomide in 53 patient plasma samples were measured,and the standardized average peak concentration of temozolomide during concurrent chemoradiation was much higher than that during adjuvant chemotherapy(8.25μg·mL^(-1) vs 5.82μg·mL^(-1)).The standardized average peak concentration in male during concurrent chemoradiation was lower than that in females(5.86μg·mL^(-1) vs 12.33μg·mL^(-1)).Conclusion The proposed method is simple,rapid,accurate,precise,and appropriate for monitoring the peak concentration of temozolomide.