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目的:建立一种全新反相高效液相色谱法测定环孢素A(CsA)血浓度的方法,并用于临床血药浓度监测。方法:以环孢素D(CsD)为内标,色谱柱采用StableBond-C_(18)柱(150 mm×4.6 mm,5μm),柱温60℃;检测波长210 nm;流动相为乙腈-甲醇-水(70:8:22)和甲醇梯度洗脱。全血样品经叔丁基甲醚单步萃取后,40℃水浴通N2吹干,残渣用甲醇溶解,在酸性条件下正己烷反提,取下清液进样。结果:CsA的线性范围为24.75～1979.95 ng·ml~(-1),回归方程为Y=0.001 498C-0.02167(r= 0.998 9),回收率为(67.39±0.05)%～(78.87±0.05)%,日内RSD为5.03%～7.81%,日间RSD为5.03%～8.31%。结论:本法结果准确,操作简便,成本低,易于质量控制,适合于临床CsA血药浓度监测及药物动力学研究。
Objective: To establish a new reversed-phase high performance liquid chromatography method for the determination of cyclosporine A (CsA) blood concentration and for the clinical blood concentration monitoring. Methods: CsD was used as internal standard. The column was equipped with StableBond-C 18 column (150 mm × 4.6 mm, 5 μm) and the column temperature was 60 ℃. The detection wavelength was 210 nm. The mobile phase consisted of acetonitrile-methanol - water (70: 8: 22) and methanol gradient. The whole blood sample was extracted with tert-butyl methyl ether in one step, and the mixture was dried in a water bath at 40 ° C. under N2. The residue was dissolved in methanol and n-hexane was back-extracted under acidic conditions. Results: The linear range of CsA was 24.75 ~ 1979.95 ng · ml -1, the regression equation was Y = 0.001 498C-0.02167 (r = 0.998 9), the recovery was (67.39 ± 0.05)% ~ (78.87 ± 0.05) %, Day RSD 5.03% ~ 7.81%, daytime RSD 5.03% ~ 8.31%. Conclusion: The method is accurate, easy to operate, low cost and easy to quality control. It is suitable for the monitoring of clinical CsA concentration and pharmacokinetics.