LC-MS/MS法测定干血斑样品中阿戈美拉汀

目的: 建立快速、灵敏的液相色谱-串联质谱(LC-MS/MS)法,测定大鼠干血斑(DBS)样品中的阿戈美拉汀,并应用于阿戈美拉汀大鼠药动学研究. 方法: 给药后的大鼠新鲜全血40 μL点于Whatman 903滤纸上,室温下干燥2 h,放入含干燥剂的密封袋保存.取直径为6 mm的干血斑3片,加入甲醇500 μL,超声15 min后,进行液-液萃取,以甲醇-甲酸铵-甲酸为流动相,流速0.20 mL·min^-1,采用Grom-Sil 120 ODS-5 ST柱分离,通过电喷雾三重四极杆串联质谱,以多反应监测(MRM)方式进行检测,用于定量的离子反应为m/z 244.1→m/z 185.1(阿戈美拉汀)和m/z 249.2→m/z 187.1(d₅-阿戈美拉汀). 结果: 测定阿戈美拉汀DBS的线性范围为0.025 0~25.0 ng·mL^-1;定量下限为0.025 0 ng·mL^-1;精密度和准确度均符合要求. 结论: DBS采样法全血用量少,样品储存、运输便利,可用于阿戈美拉汀大鼠药动学研究.

Objective: To develop and validate a liquid chromatograph/tandem mass spectrometric(LC-MS/MS)method for the determination of agomelatine in samples of dried blood spots(DBS)and to apply this method to the pharmacokinetic study in rats. Methods: An aliquot of 40 μL of whole blood sample from rats after administration was spotted onto the Whatman 903 sampling filter paper and allowed to dry at ambient temperature for 2 h.The DBS samples were stored in a sealed plastic bag containing desiccant.Three 6-mm diameter disks(equivalent to c.a.30 μL blood)were collected from the filter paper,and 500 μL of methanol was added followed by sonication for 15 min.The samples were chromatographed on a Grom-Sil 120 ODS-5 ST column.The mobile phase consisted of methanol-ammonium formate-formic acid,at a flow rate of 0.20 mL·min^-1.A tandem mass spectrometer equipped with ESI source operated in positive ionization mode was used as the detector.Multiple reaction monitoring(MRM)with the precursor/product ion combinations of m/z 244.1→m/z 185.1 amu and m/z 249.2→m/z 187.1 amu were used to quantify agomelatine and d₅-agomelatine,respectively. Results: The linear range of agomelatine in DBS was 0.025 0-25.0 ng·mL^-1 and the lower limit of quantification was 0.025 0 ng·mL^-1.The precision and accuracy results for the calibration standards and validation samples were within acceptable criteria. Conclusion: The DBS sampling offers the advantages of less sample volume and convenience of sample storage and shipment.The method can be successfully applied to the pharmacokinetic study of agomelatine in rats.