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目的:建立血浆和脑脊液中两性霉素B的HPLC测定方法。方法:色谱柱为迪马C18(5μm,150mm×4.6mm);流动相为0.005mol·L-1乙二胺四醋酸二钠-甲醇(30∶70);流速1.0mL·min-1;检测波长406nm;进样量10μL;柱温30℃。结果:血浆和脑脊液中两性霉素B的回归方程分别为C=1.054×10-4A+3.623×10-2(R=0.9997)和C=9.622×10-5A+1.91×10-3(R=0.9992),血浆中两性霉素B在0.2～10mg·L-1范围内、脑脊液中两性霉素B在0.02～1mg·L-1范围内与峰面积间均呈良好的线性关系。两个方法的日内和日间精密度均小于5%。结论:本方法灵敏、准确、重现性好,适用于两性霉素B临床药动学和药物浓度测定的需要。
Objective: To establish a HPLC method for the determination of amphotericin B in plasma and cerebrospinal fluid. METHODS: The chromatographic column was Dima C18 (5μm, 150mm × 4.6mm). The mobile phase consisted of 0.005mol·L -1 disodium ethylenediaminetetraacetate (30:70) and the flow rate was 1.0mL · min-1. Wavelength 406nm; injection volume 10μL; column temperature 30 ℃. Results: The regression equations of amphotericin B in plasma and cerebrospinal fluid were C = 1.054 × 10-4A + 3.623 × 10-2 (R = 0.9997) and C = 9.622 × 10-5A + 1.91 × 10-3 (R = 0.9992). The concentration of amphotericin B in plasma ranged from 0.2 mg / L to 10 mg · L-1, and the linear range of amphotericin B in the range of 0.02 mg / L to 1 mg · L-1 was linear with the peak area. The intra-day and inter-day precision of both methods were less than 5%. Conclusion: The method is sensitive, accurate and reproducible. It is suitable for the clinical pharmacokinetics and drug concentration determination of amphotericin B.