Gaillard Y, Prévosto J M, Cheminel V, Soares O, Chaulet J F
Laboratoire de Biochimie, Pharmacologie et Toxicologie du Pr. Blanchot, Hôpital d'Instruction des Armées Desgenettes, Lyon, France.
J Chromatogr B Biomed Appl. 1995 Jun 23;668(2):315-21. doi: 10.1016/0378-4347(95)00075-t.
A rapid, accurate, and sensitive high-performance liquid chromatographic (HPLC) method, with fluorimetric detection, for the simultaneous measurement of halofantrine and desbutylhalofantrine in human plasma or whole blood is described. Sample preparation involved protein precipitation, followed by an efficient solid-phase extraction on a C8 cartridge. Analytes were isolated from 1 ml of the biological fluids and recovered by a 2% acetic acid in ethyl acetate solution. Chromatographic separation was carried out on a LiChrospher 60 RP select B, C8 bonded phase (5 microns particle size, 25 cm x 4 mm I.D.) using a mobile phase of water-acetonitrile (35:65, v/v) containing triethylamine (1%) and adjusted to pH 4 with orthophosphoric acid. The total run time was 14 min. Relative standard deviations of the intra-and inter-assay precisions were less than 5.9%. Assumption of linearity was investigated by studying the y-residuals and by ANOVA (analysis of variance). Because of the wide range of calibration (0.1 to 2.0 microgram/ml) variances were non-homogeneous (Hartley's test) and the weighted regression line was computed in order to allow pharmacokinetic studies. Accuracy was tested using a t-statistic. Limits of decision, detection and quantification were realized from an analysis of the blanks. Application of the method to clinical specimens was demonstrated.
本文描述了一种快速、准确且灵敏的高效液相色谱（HPLC）方法，采用荧光检测，用于同时测定人血浆或全血中的卤泛群和去丁基卤泛群。样品制备包括蛋白沉淀，随后在C8柱上进行高效固相萃取。从1 ml生物流体中分离出分析物，并用2%乙酸乙酯溶液回收。色谱分离在LiChrospher 60 RP select B C8键合相（粒径5微米，25 cm×4 mm内径）上进行，流动相为水 - 乙腈（35:65，v/v），含有三乙胺（1%），并用正磷酸调节至pH 4。总运行时间为14分钟。批内和批间精密度的相对标准偏差小于5.9%。通过研究y残差和方差分析（ANOVA）来研究线性假设。由于校准范围宽（0.1至2.0微克/毫升），方差不均匀（Hartley检验），因此计算加权回归线以进行药代动力学研究。使用t统计量测试准确性。通过对空白样品的分析确定判定限、检测限和定量限。证明了该方法在临床标本中的应用。
