Cunliffe Jennifer M, Shen Jim X, Wei Xianrong, Dreyer Daniel P, Hayes Roger N, Clement Robert P
Merck Research Labs, Summit, NJ 07901, USA.
Bioanalysis. 2011 Apr;3(7):735-43. doi: 10.4155/bio.11.17.
The fast-paced nature of the pharmaceutical industry requires robust bioanalytical methods to endure the high-throughput sample demands of the production environment.
A rapid, accurate and precise LC-MS/MS method was developed for the quantitation of a diastereomer quartet in human plasma. Virtually all of the phosphatidylcholine and most of the lysophosphatidylcholine from human plasma were removed using a phospholipid-removing protein precipitation 96-well plate. An Agilent Poroshell SB-C18 2.1 × 50 mm superficially porous column was used at 100°C and 1.2 ml/min to separate a diastereomer quartet in <2.5 min. Peak shape, retention and resolution were maintained over nearly 200 extracted bioanalytical samples under these separation conditions. The method was tested for accuracy and precision; the assay inter-run accuracy and precision were minus 7.2-0.7% and 2.1-11.9%, respectively (n = 18).
The application of the superficially porous column resulted in twofold response increase and a 2.6-fold reduction in cycle time compared with a 3.5-µm column performing under comparable resolution conditions.
制药行业的快节奏特性要求有强大的生物分析方法来满足生产环境中高通量样品的需求。
开发了一种快速、准确且精密的液相色谱-串联质谱法用于定量人血浆中的非对映异构体四重峰。使用一种去除磷脂的蛋白沉淀96孔板几乎去除了人血浆中所有的磷脂酰胆碱和大部分溶血磷脂酰胆碱。在100°C和1.2 ml/min流速下使用安捷伦Poroshell SB-C18 2.1×50 mm表面多孔柱在不到2.5分钟内分离出非对映异构体四重峰。在这些分离条件下，近200个提取的生物分析样品的峰形、保留时间和分离度得以保持。对该方法进行了准确性和精密度测试；批间测定准确性和精密度分别为-7.2 - 0.7%和2.1 - 11.9%（n = 18）。
与在可比分离度条件下运行的3.5 µm柱相比，表面多孔柱的应用使响应增加了两倍，循环时间减少了2.6倍。
