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目的建立人血浆中拉米夫定(3TC)、地丹诺辛(ddI)和司他夫定(d4T)的HPLC测定方法。方法取血浆250μL,加入内标替加氟和醋酸胺缓冲液混匀,乙腈沉淀蛋白,取上清液常温下氮气吹干,残渣以流动相溶解进样。色谱柱采用Shim-packCLC-ODS(6mm×150mm,5μm)柱,流动相为:乙腈-磷酸盐缓冲液(6∶94)。0～9min紫外检测波长为260nm,9～11min紫外检测波长240nm,11～22min紫外检测波长为260nm。柱温为35℃。结果在本试验条件下,3TC在0.02～5mg·mL-1内线性良好(r=0.9996,n=6)。ddI在0.05～5mg·mL-1内线性良好(r=0.9997,n=6)。d4T在0.02～5mg·mL-1内线性良好(r=0.9996,n=6)。高、中、低3个浓度质控样品中,批内误差(RSD)为1.03%～2.70%,批间(RSD)为1.50%～6.77%。结论本方法操作简便,结果准确可靠,重现性好,为药动学和临床药物研究提供了方法学基础。
Objective To establish an HPLC method for the determination of lamivudine (3TC), ddanoxin (ddI) and stavudine (d4T) in human plasma. Methods Take 250μL of plasma, add internal standard of Teflon and ammonium acetate buffer, and mix with acetonitrile to precipitate protein. The supernatant was dried under nitrogen at room temperature and the residue was dissolved in mobile phase. The column was Shim-pack CLC-ODS (6 mm × 150 mm, 5 μm) and the mobile phase was acetonitrile-phosphate buffer (6:94). 0 ~ 9min UV detection wavelength of 260nm, 9 ~ 11min UV detection wavelength of 240nm, 11 ~ 22min UV detection wavelength of 260nm. The column temperature was 35 ° C. Results Under the experimental conditions, 3TC showed a good linearity within 0.02 ~ 5 mg · mL-1 (r = 0.9996, n = 6). The linearity of ddI was good in 0.05 ~ 5 mg · mL-1 (r = 0.9997, n = 6). The linearity of d4T was 0.02 ~ 5 mg · mL-1 (r = 0.9996, n = 6). In the high, middle and low quality control samples, the intra-assay error (RSD) ranged from 1.03% to 2.70% and the intra-assay RSD ranged from 1.50% to 6.77%. Conclusion The method is simple, accurate, reliable and reproducible. It provides a methodological foundation for the study of pharmacokinetics and clinical medicine.