March C, Karnes H T, McLean A, Mukherjee P S
Department of Pharmaceutics, Virginia Commonwealth University, Richmond, VA 23298-0533, USA.
Biomed Chromatogr. 2001 Apr;15(2):100-7. doi: 10.1002/bmc.45.
This paper describes the validation of an analytical method for the determination of amphetamine in beagle dog plasma by gas chromatography coupled to mass spectrometry (GC-MS). d-Amphetamine-d(6) was used as the internal standard. The method consisted of a rapid single-step liquid-liquid extraction and derivatization of amphetamine with 2,2,2-trichloroethyl chloroformate, followed by sensitive GC-MS detection. This is the first report utilizing the combination of trichloroethyl chloroformate as a derivatization reagent and a deuterated amphetamine analog as an IS for the quantification of amphetamine in plasma. The method was validated in terms of specificity, curve fit, precision, accuracy, recovery and stability, and was acceptable according to FDA draft guidelines for validation of bioanalytical methods. The limit of detection was 0.65 ng/mL. The calibration range was 5-150 ng/mL. The validated method was successfully employed for the quantitation of amphetamine in dog plasma samples for pharmacokinetic profiling.
本文描述了一种采用气相色谱-质谱联用（GC-MS）测定比格犬血浆中苯丙胺的分析方法的验证。d-苯丙胺-d(6)用作内标。该方法包括快速单步液-液萃取以及苯丙胺与氯甲酸2,2,2-三氯乙酯的衍生化反应，随后进行灵敏的GC-MS检测。这是首篇利用氯甲酸三氯乙酯作为衍生化试剂和氘代苯丙胺类似物作为内标对血浆中苯丙胺进行定量分析的报告。该方法在特异性、曲线拟合、精密度、准确度、回收率和稳定性方面得到了验证，并且根据FDA生物分析方法验证草案指南是可接受的。检测限为0.65 ng/mL。校准范围为5-150 ng/mL。经验证的方法成功用于狗血浆样品中苯丙胺的定量分析，以进行药代动力学分析。
