Quantitative determination of total methamphetamine and active metabolites in rat tissue by liquid chromatography with tandem mass spectrometric detection
Quantitative determination of total methamphetamine and active metabolites in rat tissue by liquid chromatography with tandem mass spectrometric detection
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DOI：
10.1208/aapsj080480
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发表时间：
2006-01-01
期刊：
影响因子：
4.5
通讯作者：
Owens, S. Michael
中科院分区：
文献类型：
--
作者：
Hendrickson, Howard;Laurenzana, Elizabeth;Owens, S. Michael
采用简单三氯乙酸样品处理，开发并验证了用于测定甲基苯丙胺（METH）、苯丙胺（AMP）、4-羟基甲基苯丙胺（4-OH-METH）和4-羟基苯丙胺（4-OH-AMP）的高通量液相色谱串联质谱检测（LC-MS/MS）方法。在大鼠血清、脑和睾丸中验证了该方法。使用正离子电喷雾串联质谱法（MS/MS）测定METH和AMP的定量下限（LOQ）为1 ng中心点mL（-1）。在广泛的分析物浓度范围内，该方法的准确度在实际值的25%以内。批内精密度优于12%（变异系数）。该方法在较宽的动态范围（0.3-1000 ng中心点mL（-1））内呈线性。由于基质相关离子效应极小或使用了内标物，因此仅使用血清标准品可在所有3种基质中进行定量。最后，使用LC-MS/MS法测定大鼠皮下输注（5.6 mg/kg/天）METH期间血清、脑和睾丸中METH和AMP的浓度。实验样品中4-OH-AMP和4-OH-METH的浓度低于LOQ。使用血清校准品测定睾丸和脑中METH和AMP浓度引入的偏倚小于8%，相对于动物间变异性而言不显著。
High-throughput liquid chromatography with tandem mass spectrometric detection (LC-MS/MS) methodology for the determination of methamphetamine ( METH), amphetamine (AMP), 4-hydroxymethamphetamine (4-OH-METH), and 4-hydroxyamphetamine (4-OH-AMP) was developed and validated using simple trichloroacetic acid sample treatment. The method was validated in rat serum, brain, and testis. Lower limits-of-quantitation (LOQ) for METH and AMP were 1 ng center dot mL(-1) using positive ion electrospray tandem mass spectrometry (MS/MS). The accuracy of the method was within 25% of the actual values over a wide range of analyte concentrations. The within-assay precision was better than 12% (coefficient of variation). The method was linear over a wide dynamic range (0.3-1000 ng center dot mL(-1)). Quantitation was possible in all 3 matrices using only serum standards because of minimal matrix-associated ion effects or the use of an internal standard. Finally, the LC-MS/MS method was used to determine serum, brain, and testis METH and AMP concentrations during a subcutaneous infusion (5.6 mg kg(-1) day(-1)) of METH in rats. Concentrations of 4-OH-AMP and 4-OH-METH were below the LOQ in experimental samples. The bias introduced by using serum calibrators for the determination of METH and AMP concentrations in testis and brain was less than 8% and insignificant relative to the interanimal variability.