高效液相色谱法测定人血浆和脑脊液中的两性霉素B

Determination of amphotericin B in human plasma and cerebrospinal fluid by HPLC

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 develop HPLC method for assaying amphotericin B in human plasma and cerebrospinal fluid. METHODS The chromatographic separation was achieved on a reverse-phase column DIKMA Kromasil C18 （150 mm × 4. 6 mm,5 μm） using 0. 005 mol·L ^-1 EDTA methanol（30：70）as mobile phase. The flow rate was 1. 0 mL·min^-1 and effluent was monitored at 4（16 am. The column temperature was 30 ℃ and sample size was 10μL. RFNULTS The linear response over the concentration ranges of 0. 2- 10 mg·L^-1 and 0.02- 1 mg·L^-1 were obtained in plasma and CSF, respectively. The linear regression equations in plasma and CSF were C= 1. 054 × 10^-4A ＋ 3. 623 × 10 ^-2 （R = 0. 999 7） and C = 9. 622 × 10^-5A ＋ 1.91 × 10^- 3 （R = 0. 999 2）. Intra-day and inter-day RSDs were below 5 % for all of the samples. CONCLUSION The method is sensitive, accurate with good reproducibility and suitable for determination of amphotericin B in human plasma and cerebrospinal fluid for clinical pharmacokinetics and drug monitoring.