Determination of didanosine in human serum by on-line solid-phase extraction coupled to high-performance liquid chromatography with electrospray ionization tandem mass spectrometric detection: application to a bioequivalence study.
Determination of didanosine in human serum by on-line solid-phase extraction coupled to high-performance liquid chromatography with electrospray ionization tandem mass spectrometric detection: application to a bioequivalence study.
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DOI：
10.1002/jms.449
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发表时间：
2003-04
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影响因子：
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通讯作者：
R. Estrela;M. Salvadori;R. Raices;G. Suarez-Kurtz
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文献类型：
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作者：
R. Estrela;M. Salvadori;R. Raices;G. Suarez-Kurtz
建立了固相萃取-液相色谱-电喷雾离子化-串联质谱法测定人血清中去羟肌苷的方法。以多反应监测模式进行采集，监测去羟肌苷的跃迁m/z 237 -> 136.7和拉米夫定的跃迁m/z 230 -> 111.7。该方法在10-1500 ng ml（-1）范围内呈线性关系，r（2）> 0.98，运行时间为5 min，批内和批间精密度为95%。绝对回收率分别为99.8%（10 ng ml（-1））、98.4%（30 ng ml（-1））、91.5%（700 ng ml（-1））和94.7%（1200 ng ml（-1））。检测限和定量限分别为5和10 ng/ml。该方法应用于生物等效性研究，其中24名健康成年志愿者（12名男性）接受单次口服剂量（200 mg）的参考和测试去羟肌苷制剂（口服溶液缓冲粉末），在一个开放的，双向的，随机的，交叉协议。C（max）（血清峰浓度）和AUC（0-inf）（从时间0至无穷大的血清浓度-时间曲线下面积）的个体比值（供试制剂/参比制剂）的90%置信区间在80- 125%范围内，这支持两种制剂在去羟肌苷吸收速率和程度方面具有生物等效性的结论。
A method based on solid-phase extraction coupled to liquid chromatography with positive ion electrospray ionization and tandem mass spectrometric detection was developed for the determination of didanosine in human serum, using lamivudine as internal standard. The acquisition was performed in the multiple reaction monitoring mode, monitoring the transitions m/z 237 --> 136.7 for didanosine and m/z 230 --> 111.7 for lamivudine. The method was linear over the range studied (10-1500 ng ml(-1)), with r(2) > 0.98, and the run time was 5 min. The intra- and inter-assay precisions were 95%. The absolute recoveries were 99.8% (10 ng ml(-1)), 98.4% (30 ng ml(-1)), 91.5% (700 ng ml(-1)) and 94.7% (1200 ng ml(-1)). The limits of detection and quantitation were 5 and 10 ng ml(-1), respectively. The method was applied to a bioequivalence study, in which 24 healthy adult volunteers (12 men) received single oral doses (200 mg) of reference and test didanosine formulations (buffered powder for oral solutions), in an open, two-way, randomized, crossover protocol. The 90% confidence interval of the individual ratios (test formulation/reference formulation) for C(max) (peak serum concentration) and AUC(0-inf) (area under the serum concentration versus time curve from time zero to infinity) were within the range 80-125%, which supports the conclusion that the two formulations are bioequivalent regarding the rate and extent of didanosine absorption.