Ayrton J, Clare R A, Dear G J, Mallett D N, Plumb R S
International Bioanalysis and Drug Metabolism Division, Glaxo Wellcome Research and Development, Park Road, Ware, Herts SG12 0DP, UK.
Rapid Commun Mass Spectrom. 1999;13(16):1657-62. doi: 10.1002/(SICI)1097-0231(19990830)13:16<1657::AID-RCM694>3.0.CO;2-L.
Ultra-high flow rate liquid chromatography on large particle size stationary phases coupled with mass spectrometric detection (particularly tandem mass spectrometry, MS/MS) is gaining increasing usage for the direct determination of pharmaceuticals in biological fluids. The lack of sample preparation required prior to chromatographic and MS/MS analysis, together with the extremely high throughput of the chromatography, make the technique extremely attractive to the modern pharmaceutical bioanalyst. However, this lack of sample preparation also means that there is no potential for concentration of the sample and, as a consequence, the sensitivity of the technique has been limited. Liquid chromatography on the capillary scale offers sensitivity benefits compared with conventional liquid chromatography as the volume in which the analyte peaks are eluted is greatly reduced. In this paper, we present the use of ultra-high flow rate liquid chromatography on the capillary scale. This enables the quantification of drugs in plasma at sub-nanogram per millilitre concentrations from a very small (2.5 micromgL) aliquot of plasma without sample preparation. We also compare the resolution obtained by ultra-high flow rate liquid chromatography with that achieved on short columns packed with conventional size packing materials operated in an isocratic manner.
大粒径固定相上的超高速液相色谱与质谱检测（特别是串联质谱，MS/MS）联用，在生物流体中药物的直接测定方面的应用越来越广泛。在色谱和MS/MS分析之前无需样品制备，再加上色谱的极高通量，使得该技术对现代药物生物分析人员极具吸引力。然而，缺乏样品制备也意味着没有浓缩样品的可能性，因此，该技术的灵敏度受到了限制。与传统液相色谱相比，毛细管规模的液相色谱具有灵敏度优势，因为分析物峰洗脱的体积大大减少。在本文中，我们展示了毛细管规模的超高速液相色谱的应用。这使得在无需样品制备的情况下，能够从非常少量（2.5微升）的血浆等分试样中定量测定血浆中浓度为亚纳克每毫升的药物。我们还将超高速液相色谱获得的分离度与在填充常规尺寸填料的短柱上以等度方式操作所实现的分离度进行了比较。
