School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
J Pharm Biomed Anal. 2012 Dec;71:104-10. doi: 10.1016/j.jpba.2012.08.012. Epub 2012 Aug 19.
A sensitive, specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for the quantitative determination of amlodipine and bisoprolol, using clenbuterol as the internal standard (IS). The analytes and IS were isolated from 100μL plasma samples by a simple liquid-liquid extraction (LLE). Reverse-phase high performance liquid chromatography (RP-HPLC) separation was accomplished on a Diamonsil C(18) column (50mm×4.6mm, 5μm) with a mobile phase composed of methanol-water-formic acid (75:25:0.01, v/v/v) at a flow rate of 0.3mL/min. The method had a chromatographic total run time of 3min. Multiple reacting monitoring (MRM) transitions of m/z M+H 409.1→237.9 (amlodipine), m/z M+H 326.2→116.0 (bisoprolol) and m/z M+H 277.0→203.0 (clenbuterol, IS) were used to quantify amlodipine, bisoprolol and IS, respectively. The method was sensitive with a lower limit of quantitation (LLOQ) of 0.2ng/mL for both amlodipine and bisoprolol, and the linear range was 0.2-50ng/mL for both amlodipine and bisoprolol (r(2)>0.9961). All the validation data, such as accuracy, precision and inter-day repeatability, were within the required limits. The method was successfully applied to pharmacokinetic studies of amlodipine and bisoprolol in Sprague-Dawley (SD) rats.
建立了一种灵敏、特异的液相色谱-串联质谱（LC-MS/MS）法，用于定量测定氨氯地平和比索洛尔，以克仑特罗为内标（IS）。分析物和 IS 从 100μL 血浆样品中通过简单的液液萃取（LLE）分离。反相高效液相色谱（RP-HPLC）分离在 Diamonsil C（18）柱（50mm×4.6mm，5μm）上进行，流动相由甲醇-水-甲酸（75:25:0.01，v/v/v）组成，流速为 0.3mL/min。该方法的总色谱运行时间为 3min。采用多反应监测（MRM）跃迁 m/z [M+H]（+）409.1→237.9（氨氯地平）、m/z [M+H]（+）326.2→116.0（比索洛尔）和 m/z [M+H]（+）277.0→203.0（克仑特罗，IS）分别定量测定氨氯地平、比索洛尔和 IS。该方法具有较高的灵敏度，氨氯地平和比索洛尔的定量下限（LLOQ）均为 0.2ng/mL，线性范围分别为 0.2-50ng/mL（r（2）>0.9961）。所有验证数据，如准确度、精密度和日间重复性，均在要求范围内。该方法成功应用于 Sprague-Dawley（SD）大鼠中氨氯地平和比索洛尔的药代动力学研究。
