Department of Bioanalytical and Discovery Analytical Science, Bristol-Myers Squibb Co., Princeton, NJ 08543-5400, USA.
J Sep Sci. 2010 Mar;33(6-7):923-9. doi: 10.1002/jssc.200900659.
RP-HPLC has largely been the analytical method of choice in the pharmaceutical industry for many years because of the poor aqueous solubility and hydrophobicity of most small molecule drug candidates. RP-HPLC coupled to MS has provided an excellent analytical tool for qualitatively and quantitatively determining levels of drug molecules or drug metabolites for studies such as purity assessment and pharmacokinetic profiling. Quantitation of endogenous metabolites is an emerging field in drug discovery, gaining popularity for evaluating pharmacokinetic-pharmacodynamic relationships and targeting activity and efficacy. While RP-HPLC-MS is suitable for a range of applications, many endogenous molecules, especially those found in urine, are small polar compounds that do not retain well by RP-HPLC. This has made hydrophilic interaction LC (HILIC) an attractive alternative and useful approach to polar molecule analysis. Additionally, because HILIC is routinely used with traditional RP organic solvents such as ACN and methanol, it can be easily coupled to MS. This paper will review selected examples from the current literature as well as discuss some new applications from the author's own laboratory focusing on the applicability of HILIC to quantitative and qualitative profiling of endogenous metabolites in drug discovery.
多年来，由于大多数小分子药物候选物的水溶性和疏水性差，反相高效液相色谱（RP-HPLC）在制药行业中基本上一直是首选的分析方法。RP-HPLC 与 MS 相结合，为定性和定量测定药物分子或药物代谢物的水平提供了极好的分析工具，例如用于纯度评估和药代动力学分析。内源性代谢物的定量分析是药物发现中的一个新兴领域，越来越受欢迎，用于评估药代动力学-药效学关系以及靶向活性和功效。虽然 RP-HPLC-MS 适用于多种应用，但许多内源性分子，尤其是尿液中发现的那些，都是极性小的化合物，用 RP-HPLC 保留效果不佳。这使得亲水作用色谱（HILIC）成为一种很有吸引力的替代方法和极性分子分析的有用方法。此外，由于 HILIC 通常与传统的 RP 有机溶剂（如乙腈和甲醇）一起使用，因此可以很容易地与 MS 结合。本文将综述当前文献中的一些精选实例，并讨论作者自己实验室的一些新应用，重点介绍 HILIC 在药物发现中对内源性代谢物的定量和定性分析中的适用性。
