目的 建立高效液相色谱串联质谱(HPLC-MS/MS)直接检测唾液中氨磷汀(amifostine,AMI)的方法。方法 收集6位健康成年志愿者唾液为基质,石杉碱甲（huperzine-A,HupA）为内标,采用蛋白沉淀法,用高效液相色谱串联质谱测定唾液中的氨磷汀。色谱柱为ZIC-HILIC亲水色谱分析柱（2.1 mm×100 mm,3.5 μm）,质谱分析采用多反应监测扫描模式（MRM）,离子源为电喷雾离子源（ESI）。结果 氨磷汀在唾液中检测到的线性范围为0.938～30 mg·L-1 ,线性关系良好,典型代表方程为：y=0.072x-0.005 26 (r=0.999 1) (n=6),定量下限为0.938 mg·L-1(S/N>10)。唾液样品的低、中、高3个质控浓度（1.0、5.0和25 mg·L-1）批内、批间精密度(RSD)均小于15%,其方法回收率均大于85%。结论 本实验所建立的氨磷汀检测方法直接、快捷、简便、灵敏度高及定性好,适用于唾液中氨磷汀的测定及科学研究。
Abstract
Objective To establish an HPLC-MS/MS method (high performance liquid chromatography-tandem mass spectrometry) for the direct determination of amifostine in human saliva. METHODS Saliva samples were collected from six adult healthy volunteers. After protein precipitation and addition of the internal standard (IS) huperzine-A (HupA),HPLC- MS/MS was used to analyze amifostine. The analysis was conducted using a ZIC-HILIC analytical column (2.1 mm×100 mm,3.5 μm). Electrospray ionization was used with multiple reaction monitoring (MRM) mode. RESULTS The lower limit of quantification (LLOQ) of the method was 0.938 mg·L-1 (S/N>10). The standard curve was linear in the range of 0.938-30 mg·L-1 (r=0.999 1,n=6). The inter-day and intra-day RSDs were all less than 15% for the low,medium and high concentration quality control samples (1.0,5.0 and 25 mg·L-1 ). The values of recovery were all more than 85%. CONCLUSION The method is direct,rapid,simple and sensitive,and suitable for the determination of amifostine in saliva samples.
关键词
高效液相色谱质谱联用法 /
氨磷汀 /
唾液
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Key words
HPLC -MS/MS /
amifostine /
saliva
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中图分类号：
R917
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