Hotha Kishore Kumar, Roychowdhury Swapan, Mullangi Ramesh, Ravindranath L K
Analytical Research and Development, Novel Laboratories Inc., Somerset, NJ 08873, USA.
Biomed Chromatogr. 2013 Sep;27(9):1192-9. doi: 10.1002/bmc.2926. Epub 2013 May 3.
A rapid, simple, specific and sensitive LC-MS/MS method has been developed and validated for the enantiomeric quantification of amlodipine (AML) isomers [R-amlodipine (R-AML) and S-amlodipine (S-AML)] with 200 μL of human plasma using R-AML-d4 and S-AML-d4 as corresponding internal standards as per regulatory guidelines. A simple liquid-liquid extraction process was used to extract these analytes from human plasma. The total run time was 3.5 min and the elution of R-AML, S-AML, R-AML-d4 and S-AML-d4 occurred at 1.62, 2.51, 1.63 and 2.53 min, respectively. This was achieved with a mobile phase consisting of 0.2% ammonia-acetonitrile (20:80, v/v) at a flow rate of 1 mL/min on a Chiralcel OJ RH column. A linear response function was established for the range of concentrations 0.1-10 ng/mL (r >0.998) for each enantiomer. The intra- and inter-day precision values for both enantiomers met the acceptance criteria. Both enantiomers were stable in a set of stability studies, viz. bench-top, auto-sampler, freeze-thaw cycles and long-term. The current assay was successfully applied to a pharmacokinetic study to quantitate AML enantiomers following oral administration of 10 mg AML tablet to humans.
已开发并验证了一种快速、简单、特异且灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于使用R-氨氯地平-d4和S-氨氯地平-d4作为相应内标，对200 μL人血浆中的氨氯地平（AML）异构体[R-氨氯地平（R-AML）和S-氨氯地平（S-AML）]进行对映体定量，该方法符合监管指南。采用简单的液-液萃取过程从人血浆中提取这些分析物。总运行时间为3.5分钟，R-AML、S-AML、R-AML-d4和S-AML-d4的洗脱时间分别为1.62、2.51、1.63和2.53分钟。这是在Chiralcel OJ RH柱上，以0.2%氨-乙腈（20:80，v/v）为流动相，流速为1 mL/min的条件下实现的。为每种对映体在0.1-10 ng/mL的浓度范围内建立了线性响应函数（r>0.998）。两种对映体的日内和日间精密度值均符合验收标准。在一系列稳定性研究中，即台式、自动进样器、冻融循环和长期稳定性研究中，两种对映体均稳定。当前的分析方法已成功应用于一项药代动力学研究，以定量人类口服10 mg AML片剂后AML对映体的含量。
