高效液相色谱High-performance liquid chromatography

最常用的是反相色谱：固定相为非极性的 C18（十八烷基）键合硅胶，流动相为水与甲醇或乙腈的混合物，极性化合物先流出、非极性化合物后流出。正相色谱用极性固定相和非极性流动相；还有离子交换色谱、尺寸排阻色谱（测定高分子分子量）和手性色谱。可用等度洗脱或梯度洗脱。检测器包括紫外-可见/二极管阵列（最常用）、荧光、示差折光、蒸发光散射和质谱。

Reversed phase is most common: a non-polar C18 (octadecyl) bonded silica with a mobile phase of water mixed with methanol or acetonitrile, so polar compounds elute first and non-polar ones last. Normal phase uses a polar stationary phase and non-polar mobile phase, and there are also ion-exchange, size-exclusion (for polymer molar mass) and chiral modes, run isocratically or with gradients. Detectors include UV–visible or diode-array (the most common), fluorescence, refractive index, evaporative light scattering and mass spectrometry.

1960 年代末霍瓦斯、科克兰和胡贝尔等发展了高压液相色谱，1970 年代反相键合固定相普及。颗粒越小柱效越高但压力越大：2004 年沃特斯公司推出超高效液相色谱（UPLC，1.7 μm 颗粒，压力可达 100 MPa 以上），分析时间缩短到几分钟。核-壳型颗粒以较低压力实现了类似的高效。

In the late 1960s Horváth, Kirkland, Huber and others developed high-pressure liquid chromatography, and reversed-phase bonded phases spread in the 1970s. Smaller particles give higher efficiency at higher pressure: in 2004 Waters introduced UPLC with 1.7 μm particles and pressures above 100 MPa, cutting runs to minutes. Core–shell particles achieve similar efficiency at lower pressure.

HPLC 用于药物含量和有关物质测定（各国药典的主要方法）、生物样品中药物浓度监测、氨基酸和蛋白质多肽分析、维生素和食品添加剂检测、中药指纹图谱、环境中多环芳烃和农药分析，以及制备型 HPLC 纯化天然产物和药物。与质谱联用（LC-MS/MS）是代谢组学和蛋白质组学的核心技术。

HPLC assays drug content and related substances (the main method in pharmacopoeias), monitors drug levels in biological samples, analyses amino acids, peptides and proteins, vitamins and food additives, fingerprints herbal medicines, measures polycyclic aromatic hydrocarbons and pesticides in the environment, and in preparative form purifies natural products and drugs. Coupled with tandem mass spectrometry (LC-MS/MS) it is central to metabolomics and proteomics.