HPLC-MS／MS法测定人体血清中替加色罗的浓度

HPLC-MS/MS determination of tegaserod concentration in human serum

目的:建立一种测定人体血浆中替加色罗血药浓度的液相色谱-质谱/质谱的方法。方法:色谱柱:Liehrospher C_(18)(150mm×4.6 mm,5μm),流动相:甲醇-水(含2.5 mmol·L^(-1)醋酸铵,1%甲酸)(75:25),流速:0.5 mL·min^(-1),柱温:25℃。内标为苯海拉明。质谱条件为电喷雾电离源(ESI),选择正离子检测,营加色罗:[M+H]^+离子 m/z 302.2,碎片子离子 m/z173.1,碰撞能量28 V;内标:苯海拉明[M+H]^-离子 m/z 256.2,碎片子离子 m/z 165.1,碰撞能量36 V。结果:本法线性范围:0.0361～7·215 ng·mL^(-1),r=0.9991(n=5),最低定量浓度为0.0361 ng·mL^(-1);方法的准确度在90%～115%之间;提取回收率(绝对回收率)>70%。批内、批间 RSD<10%。结论:本方法专属性强,灵敏度高,线性关系良好,适用于人体血浆中替加色罗的测定及药代动力学研究。

Objective ： To establish an HPLC - MS/MS method for the determination of tegaserod in human serum. Method：The HPLC -MS/MS method was performed on a colume of Lichrospher C18 （150 mm ×4. 6 mm,5 μm）. The mobile phase consisted of methanol - water（ contained 2.5 mmol·L^-1 CH3 COONH4 and 1% HCOOH） （75： 25） at a flow rate of 0.5 mL ·min^-1. The temperature of column was 25 ℃. The internal standard was diphen- hydramine. ESI was applied and operated in positive mode. The parent ion of tegaserod and diphenhydramine [ M ＋ H ]^＋ m/z were 302.2 and 256.2, respectively,while the product ion for tegaserod and diphenhydramine were 173.1 and 165.1 ,respectively. Results：The linear range was 0. 0361 -7. 215 ng·mL^-1 ,r =0. 9991 （n =5）. The limit of quantification was 0. 0361 ng·mL^-1. The method recovery was 90% - 115%, extracted recovery（ absolute recovery ） was more than 70% ; The intra - day precision （RSD） and inter - day precision （RSD） were less than 10%. Conclusion：The method is specific, sensitive, and accurate, and suitable for the determination of tegaserod concentration in human serum and pharmacokinetic studies.