Koks C H, Rosing H, Meenhorst P L, Bult A, Beijnen J H
Department of Pharmacy, Slotervaart Hospital, Amsterdam, Netherlands.
J Chromatogr B Biomed Appl. 1995 Jan 20;663(2):345-51. doi: 10.1016/0378-4347(94)00445-b.
A high-performance liquid chromatographic (HPLC) assay has been developed for the determination of the antifungal drug fluconazole in saliva and plasma of patients infected with the human immunodeficiency virus (HIV). Samples can be heated at 60 degrees C for 30 min to inactivate the virus without loss of the analyte. The sample pretreatment involves a liquid-liquid extraction with chloroform-1-propanol (4:1, v/v). The chromatographic analysis is performed on a Lichrosorb RP-18 (5 microns) column by isocratic elution with a mobile phase of 0.01 M acetate buffer (pH 5.0)-methanol (70:30, v/v) and ultraviolet (UV) detection at 261 nm. The lower limit of is 100 ng/ml in plasma (using 500-microliters samples) and 1 microgram/ml in saliva (using 250-microliters samples) and the method is linear up to 100 micrograms/ml in plasma and saliva. At a concentration of 5 micrograms/ml the within-day and between-day precision in plasma are 7.1 and 5.7%, respectively. In saliva the within-day and between-day precision is 10.8% (at 5 micrograms/ml). The methodology is now being used in pharmacokinetic studies in HIV-infected patients in our hospital.
已开发出一种高效液相色谱（HPLC）分析法，用于测定感染人类免疫缺陷病毒（HIV）患者唾液和血浆中的抗真菌药物氟康唑。样品可在60℃加热30分钟以使病毒失活，而分析物不会损失。样品预处理包括用氯仿 - 1 - 丙醇（4:1，v/v）进行液 - 液萃取。色谱分析在Lichrosorb RP - 18（5微米）柱上进行，采用0.01 M醋酸盐缓冲液（pH 5.0） - 甲醇（70:30，v/v）的流动相进行等度洗脱，并在261 nm处进行紫外（UV）检测。血浆中的下限为100 ng/ml（使用500微升样品），唾液中的下限为1微克/ml（使用250微升样品），该方法在血浆和唾液中线性范围高达100微克/ml。在浓度为5微克/ml时，血浆中日内和日间精密度分别为7.1%和5.7%。在唾液中，日内和日间精密度为10.8%（在5微克/ml时）。该方法目前正在我院用于HIV感染患者的药代动力学研究。
