HPLC荧光和质谱两种检测器测定大鼠血浆及组织中的表阿霉素

Determination of Epirubicin in rat plasma and tissue by HPLC with fluor-detection and MS/MS-detection

目的建立测定大鼠血浆及组织中表阿霉素浓度的方法。方法采用RP—HPLC荧光和质谱两种检测器，Luna C18（2）分析柱（150mm×4．6mm，5μm），流动相为0．01mol·L^-1乙酸铵-乙腈（60：40，甲酸调pH3．5），流速0．6ml·min^-1。样品在碱性条件下，用乙酸乙酯漩涡混合提取浓集后进样，λEx=450nm，λEx=530nm，质谱以MRM模式测定，内标法定量。结果荧光检测标准曲线在2．44～2．50×10^7μg·L^-1有良好线性，定量限2．44μg·L^-1，日内RSD小于5．0％，日间RSD小于8.6％，方法回收率99％～113％，萃取回收率86．8％～89．7％；质谱检测标准曲线在0．49—2．00×10^3μg·L^-1有良好线性，定量限0．49μg·L^-1，日内RSD小于6．5％，日间RSD小于8．7％，方法回收率98％～115％。结论所建方法快速简便、灵敏准确，适用于血浆及组织中表阿霉素的测定及药物动力学研究。

OBJECTIVE To establish an HPLC with fluor and mass - detector for the determination of epirubicin （EB） in rat plasma and tissue. METHODS Luna C18 （2） analysis column （ 150 mm × 4.6 mm, 5 μm） was used. The alkalinized sample was extracted with 3.5 ml ethyl acetate. After evaporation of the organic layer, the residue was dissolved in mobile phase and injected onto the column. The mobile phase of 0.01 mol·L^-1 ammonium acetate - acetonitrile （60 ： 40,adjusted pH to 3.5 with formic acid） was pumped at 0. 6 ml· min^ -1 through the column. The fluor - detector was set at λEm 450 nm, λEm 530 nm. RESULTS With fluor - detection, the standard curve was linear over the concentration range of 2. 44 -2. 50 × 103 μg· L^-1. The lowest concentration of detection in plasma was 2. 44 μg· L^-1, the method recovery was 99% -113% , the intra- day RSD was less than 8.6% ; inter -day RSD was less than 5.0%. With MS/MS Detection. The standard curve was linear over the concentration range of 0. 49 - 2. 00 × 103 μg· L^-1. The lowest concentration of detection in plasma was 0, 49 μg· L^-1, the method recovery was 98% - 115%, the intra - day RSD was less than 8.7%, inter - day RSD was less than 6. 5%. CONCLUSION The method with fluor and mass - detection is found to be simple, rapid, sensitive and accurate for determination of EB in rat plasma and tissue.