液相色谱-串联质谱法测定犬血管组织中雷帕霉素的含量

目的建立犬血管组织中雷帕霉素含量测定的液相色谱-串联质谱方法。方法采用AgilentZorbax C18(2.1 mm×100 mm,3.5μm)色谱柱,流动相为甲醇-水(含10 mmol.L-1醋酸铵,0.08%甲酸)(85∶15,V/V),流速为0.2 mL.min-1;电喷雾离子源,负离子电离模式,选择性反应监测扫描方式,雷帕霉素和罗红霉素(内标)检测的离子对分别为m/z 936.0→m/z 409.0和m/z 837.0→m/z 679.2。结果雷帕霉素在1～1 500μg.L-1浓度范围内有良好的线性关系(r=0.997 6),最低检测限为0.3μg.L-1,日内、日间精密度(RSD)均小于6.7%,提取回收率在81.8%～86.3%范围内,相对回收率在104.4%～109.8%范围内。结论本法简便、快速、灵敏、准确,可用于测定血管组织中雷帕霉素的含量。

Determination of rapamycin in blood vessel of dogs by LC-MS/MS

AIM To establish a LC-MS/MS method for the determination of rapamycin in blood vessel tissues of dogs.METHODS The chromatographic separation was performed on Agilent Zorbax C18(2.1 mm × 100 mm,3.5 μm)column,with a mobile phase of methanol-10mmol·L-1ammonium acetate and 0.08% formic acid containing water(85 ∶ 15,V/V),at a flow rate of 0.2 mL·min-1.Negative electrospray ionization(ESI)was employed as the ionization source.Rapamycin and internal standard(roxithromycin)were detected by use of single reaction monitoring(SRM)mode(m/z 936.0 →m/z 409.0 and m/z 837.0 →m/z 679.2,respectively).RESULTS The linear concentration range for rapamycin was 1-1 500 μg·L-1(r = 0.997 6).The limit of quantitation was 0.3 μg·L-1.Within-day RSD and between-day RSD were all less than 6.7%,the extraction recovery rate was about 81.8%-86.3% and the relative recovery were all within 104.4%-109.8%.CONCLUSION The method is simple,fast,sensitive and accurate,and is suitable for the determination of rapamycin in blood vessel tissues.