Nirogi Ramakrishna V S, Kandikere Vishwottam N, Mudigonda Koteshwara, Shukla Manoj, Maurya Santosh
Biopharmaceutical Research, Suven Life Sciences Ltd, Serene Chambers, Road 7, Banjara Hills, Hyderabad 500034, India.
Biomed Chromatogr. 2006 Sep;20(9):833-42. doi: 10.1002/bmc.600.
A simple, sensitive and rapid high-performance liquid chromatography/electrospray ionization tandem mass spectrometry method was developed and validated for the assay of amlodipine in human plasma. Following liquid-liquid extraction, the analytes were separated using an isocratic mobile phase on a reverse-phase C(18) column and analyzed by MS in the multiple reaction monitoring mode using the respective [M+H]+ ions, m/z 409/238 for amlodipine and m/z 409/228 for the IS. The assay exhibited a linear dynamic range of 50-10,000 pg/mL for amlodipine in human plasma. The lower limit of quantification was 50 pg/mL with a relative standard deviation of less than 8%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The average absolute recoveries of amlodipine and the IS from spiked plasma samples were 74.7 +/- 4.6 and 72.1 +/- 2.0%, respectively. A run time of 1.5 min for each sample made it possible to analyze more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies. The observed maximum plasma concentration (Cmax) of amlodipine (2.5 mg oral dose) was 1425 pg/mL, time to observed maximum plasma concentration (Tmax) was 8.1 h and elimination half-life (T(1/2)) was 50.1 h.
建立了一种简单、灵敏且快速的高效液相色谱/电喷雾电离串联质谱法，并对其进行了验证，用于测定人血浆中的氨氯地平。液-液萃取后，使用等度流动相在反相C(18)柱上分离分析物，并采用多反应监测模式进行质谱分析，分别使用各自的[M+H]+离子，氨氯地平的质荷比为409/238，内标的质荷比为409/228。该测定法在人血浆中氨氯地平的线性动态范围为50 - 10,000 pg/mL。定量下限为50 pg/mL，相对标准偏差小于8%。在标准曲线范围内的浓度获得了可接受的精密度和准确度。加标血浆样品中氨氯地平和内标的平均绝对回收率分别为74.7±4.6%和72.1±2.0%。每个样品的运行时间为1.5分钟，使得每天能够分析400多个血浆样品。该验证方法已成功用于分析人血浆样品，以用于药代动力学、生物利用度或生物等效性研究。观察到氨氯地平（口服剂量2.5 mg）的最大血浆浓度（Cmax）为1425 pg/mL，达到最大血浆浓度的时间（Tmax）为8.1小时，消除半衰期（T(1/2)）为50.1小时。
