Knebel N G, Grieb S, Winkler M, Locher M, van der Vlis E, Verheij E R
Department of Biological Research Biochemistry, ASTA Medica AG, Frankfurt, Germany.
J Chromatogr B Biomed Sci Appl. 1999 Jan 22;721(2):257-69. doi: 10.1016/s0378-4347(98)00469-1.
An HPLC assay with tandem mass spectrometric detection in the positive-ion Turbo-Ion-Spray (TISP) mode for the fast and sensitive determination of perifosine ((I), D-21266) in human plasma was developed, utilising the structural analogue, miltefosine ((II), D-18506), as internal standard. Automated solid-phase extraction of diluted plasma samples, based on 250-microl plasma aliquots, at pH 6.5, allowed a reliable quantification of perifosine down to 4 ng/ml. Injection of 200 microl of plasma extracts onto a 100x3 mm normal-phase analytical column at a flow-rate of 0.5 ml/min provided retention-times of 2.4 and 2.1 min for perifosine (I) and the internal standard (II), respectively. The standard curves were linear from 4 to 2000 ng/ml using weighted linear regression analysis (1/Y2). The inter-assay and intra-assay accuracies for the calibration standards were within +0.9% and -0.2%, exhibiting precisions (C.V.) of +/-6.5 and +/-7.3%, respectively. Up to 100 unknowns may be analysed each 24 h per analyst.
建立了一种采用串联质谱检测的高效液相色谱法，以结构类似物米替福新（(II)，D - 18506）为内标，在正离子涡轮离子喷雾（TISP）模式下快速灵敏地测定人血浆中哌立福新（(I)，D - 21266）。基于250微升血浆等分试样，在pH 6.5条件下对稀释后的血浆样品进行自动固相萃取，可可靠地定量检测低至4 ng/ml的哌立福新。以0.5 ml/min的流速将200微升血浆提取物注入100×3 mm正相分析柱，哌立福新（I）和内标（II）的保留时间分别为2.4分钟和2.1分钟。使用加权线性回归分析（1/Y2），标准曲线在4至2000 ng/ml范围内呈线性。校准标准品的批间和批内准确度分别在+0.9%和 - 0.2%以内，精密度（变异系数）分别为±6.5%和±7.3%。每位分析人员每24小时最多可分析100个未知样品。
