Shen Jim X, Merka Eric A, Dreyer Daniel P, Clement Robert P, Hayes Roger N
Schering-Plough Research Institute, Summit, NJ 07901, USA.
J Sep Sci. 2008 Feb;31(2):242-54. doi: 10.1002/jssc.200700401.
A LC-MS/MS method using a LC column packed with sub-2 micron particles and elevated column temperatures was validated for the quantitation of SCH 503034 diastereomers (SCH 534128 and SCH 534129) in human plasma. The method was validated over the concentration range of 2.5 to 1250 ng/mL. Inter-assay precision, based on percent relative deviation for n = 18 replicate quality controls, was 4.5% for SCH 534128 and 4.9% for SCH 534129. Inter-assay accuracy based on n = 18 replicate quality controls was +/- 7.8% for both SCH 534128 and SCH 534129. The method involved the novel application of ion pairing reagents to increase the stereoselectivity of the separation. Temperature, types of ion pairing reagent, and concentration of ion pairing reagent were all found to play significant roles in the resolution of the SCH 534128 and SCH 534129 diastereomers on a LC column packed with sub-2 micron particles. Specifically, a sensitivity increase of five-fold was demonstrated by increasing the column temperature. Without sacrificing resolution, the run time was significantly shortened when the column temperature was elevated to 100 degrees C.
一种使用填充有亚 2 微米颗粒的液相色谱柱和升高柱温的液相色谱 - 串联质谱（LC-MS/MS）方法，已针对人血浆中 SCH 503034 非对映异构体（SCH 534128 和 SCH 534129）的定量进行了验证。该方法在 2.5 至 1250 ng/mL 的浓度范围内得到验证。基于 n = 18 次重复质量控制的相对偏差百分比计算的批间精密度，对于 SCH 534128 为 4.5%，对于 SCH 534129 为 4.9%。基于 n = 18 次重复质量控制计算的批间准确度，对于 SCH 534128 和 SCH 534129 均为 +/- 7.8%。该方法涉及离子对试剂的新应用，以提高分离的立体选择性。温度、离子对试剂的类型以及离子对试剂的浓度，都被发现对填充有亚 2 微米颗粒的液相色谱柱上 SCH 534128 和 SCH 534129 非对映异构体的分离起着重要作用。具体而言，通过提高柱温证明灵敏度提高了五倍。当柱温升高到 100℃时，在不牺牲分离度的情况下，运行时间显著缩短。
