Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P. O. Box 2457, Riyadh, 11451, Saudi Arabia; Department of Analytical Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt.
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P. O. Box 2457, Riyadh, 11451, Saudi Arabia.
J Pharm Biomed Anal. 2021 Jul 15;201:114108. doi: 10.1016/j.jpba.2021.114108. Epub 2021 Apr 29.
A novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the rapid and sensitive enantioselective analysis of verapamil (VER) in rat plasma was developed and validated using new superficially porous silica isopropyl-cyclofructan 6 chiral column (LarihcShell-P, LSP). The isocratic mobile phase composed of acetonitrile: trifluoroacetic acid: 10 mM ammonium formate (100 : 0.1 : 0.1, v/v/v) at a flow rate of 0.3 mL/min was applied. Sulpride was utilized as the internal standard (IS). Positive multiple reaction monitoring (MRM) mode was used for mass spectrometry analysis, and the process of analysis was run for 5.2 min. The (S)-(-)- and (R)-(+)-VER enantiomers with the IS were extracted from plasma by using solid-phase extraction (SPE) procedure before the analysis. The C18 cartridge gave good recovery rates for both enantiomers without interference from plasma endogenous. The developed assay was successfully validated following the US-FDA guidelines. The method was linear over concentration ranges of 0.5-500 ng/mL (r ≥ 0.997) for each enantiomer (plasma). The lower limits of quantification (LLOQ) for both isomers were 0.5 ng/mL. The intra- and inter-day relative standard deviations (RSD) were less than 8.7 % and the recoveries of (S)-(-)- and (R)-(+)-VER at three spiked levels of 1.5, 250.0 and 450.0 ranged from 92.0%-98.6%. The developed assay was effectively applied in monitoring the stereoselective pharmacokinetic study of VER enantiomers in rat plasma following oral administration of racemic VER. The pharmacokinetic parameters revealed that (S)-(-)-VER demonstrated prominently higher C and AUC values than (R)-(+)-enantiomer. The newly developed approach is the first chiral LC-MS/MS for the quantification of (S)-(-)- and (R)-(+)-VER utilizing superficially porous silica isopropyl-cyclofructan 6 chiral column in rat plasma after SPE.
建立并验证了一种新型的液相色谱-串联质谱（LC-MS/MS）方法，用于快速灵敏地分析大鼠血浆中的维拉帕米（VER）对映体。该方法采用新型的多孔硅胶异丙基环糊精 6 手性柱（LarihcShell-P，LSP），以乙腈：三氟乙酸：10mM 甲酸铵（100：0.1：0.1，v/v/v）为等度流动相，流速为 0.3mL/min。舒必利被用作内标（IS）。采用正离子多反应监测（MRM）模式进行质谱分析，分析过程耗时 5.2 分钟。在分析之前，通过固相萃取（SPE）程序从血浆中提取出与 IS 结合的（S）-（-）-和（R）-（+）-VER 对映体。C18 小柱对两种对映体均具有良好的回收率，且不受内源性血浆的干扰。该方法按照美国 FDA 指南进行了成功验证。该方法在每个对映体（血浆）的浓度范围为 0.5-500ng/mL（r≥0.997）时呈线性。两种异构体的定量下限（LLOQ）均为 0.5ng/mL。日内和日间相对标准偏差（RSD）均小于 8.7%，在三个加标水平（1.5、250.0 和 450.0ng/mL）下，（S）-（-）-和（R）-（+）-VER 的回收率在 92.0%-98.6%范围内。该方法成功地应用于监测大鼠口服消旋 VER 后 VER 对映体的立体选择性药代动力学研究。药代动力学参数表明，（S）-（-）-VER 的 C 和 AUC 值明显高于（R）-（+）-对映体。该新方法是利用表面多孔硅胶异丙基环糊精 6 手性柱在 SPE 后首次在大鼠血浆中定量分析（S）-（-）-和（R）-（+）-VER 的手性 LC-MS/MS 方法。
