Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing 210009, PR China.
Anal Chim Acta. 2010 Apr 1;664(1):40-8. doi: 10.1016/j.aca.2010.02.003. Epub 2010 Feb 11.
This paper describes a novel method that combines dansyl chloride (DNS-CL) derivatization with high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS) for the sensitive and selective determination of piperazine phosphate in human plasma. After addition of ondansetron hydrochloride as internal standard (IS), piperazine phosphate was derivatized and then extracted with ethyl acetate. After being evaporated and reconstituted, the sample was analyzed using LC-ESI/MS/MS. Separation was achieved using an Agilent ZORBAX SB-C(18) (150 mm x 2.1 mm I.D., 3.5 microm) column and isocratic elution with 10 mM ammonium acetate solution (pH 3.0)-methanol (50: 50, v/v). Detection was performed on a triple-quadrupole mass spectrometer utilizing electrospray ionization (ESI) interface operating in positive ion and selected reaction monitoring (SRM) mode with the precursor to product ion transitions m/z 320-->171 for DNS-CL-piperazine phosphate and m/z 294-->170 for the IS. The method was fully validated for its selectivity, sensitivity, linearity, precision, accuracy, recovery, matrix effect and stability. The coefficient (r) of piperazine phosphate with a linear range of 0.1-15 microg mL(-1) was 0.9974-0.9995. The limit of detection and lower limit of quantification in human plasma were 0.01 and 0.1 microg mL(-1), respectively. The validated LC-ESI/MS/MS method has been successfully applied to a bioequivalence study of piperazine phosphate trochiscus in Chinese healthy male volunteers.
本文描述了一种将丹磺酰氯（DNS-CL）衍生化与高效液相色谱-电喷雾串联质谱（LC-ESI/MS/MS）相结合的新方法，用于灵敏、选择性地测定人血浆中的哌嗪磷酸盐。在加入昂丹司琼盐酸盐作为内标（IS）后，哌嗪磷酸盐进行衍生化，然后用乙酸乙酯提取。蒸发并重新配制后，采用 LC-ESI/MS/MS 分析样品。采用 Agilent ZORBAX SB-C(18)（150mm×2.1mm I.D.，3.5μm）柱，以 10mM 乙酸铵溶液（pH3.0）-甲醇（50:50，v/v）等度洗脱进行分离。采用电喷雾电离（ESI）接口的三重四极杆质谱仪进行检测，以正离子模式和选择反应监测（SRM）模式进行检测，DNS-CL-哌嗪磷酸盐的前体到产物离子过渡 m/z320→171，IS 的 m/z294→170。该方法对选择性、灵敏度、线性、精密度、准确度、回收率、基质效应和稳定性进行了全面验证。哌嗪磷酸盐的线性范围为 0.1-15μg mL(-1)，相关系数（r）为 0.9974-0.9995。人血浆中的检测限和定量下限分别为 0.01 和 0.1μg mL(-1)。验证后的 LC-ESI/MS/MS 方法已成功应用于中国健康男性志愿者哌嗪磷酸盐trochiscus 的生物等效性研究。
