Department of Bioanalysis, Experimental Medicine & Bioanalysis, Clinical & Non-Clinical R & D, Ferring Pharmaceuticals A/S, Kay Fiskers Plads 11, 2300 Copenhagen S, Denmark.
J Pharm Biomed Anal. 2010 Nov 2;53(3):537-45. doi: 10.1016/j.jpba.2010.03.024. Epub 2010 Mar 27.
Plasma concentrations after administration of peptide drugs are often low due to the high potency often seen with this class of compounds. In this work a bioanalytical method based on coupled column liquid chromatography-tandem mass spectrometry (LC-MS/MS) is presented for quantification of a peptide drug, FE 202158, under clinical development. A volume of 0.5 ml human plasma is solid phase extracted on a weak cationic exchanger. After evaporation of the solvent to dryness, the reconstituted sample is injected into a coupled column liquid chromatography system. A heart-cut from the initial column, a cyano column, is trapped on a C(4) column and thereafter injected into a microbore C(18) column. For the detection a triple quadrupole mass spectrometer, equipped with a TurboIonSpray interface working in positive ion mode, is used. The design of the system is described and the gain in sensitivity and selectivity, compared to a conventional system, is discussed. Data from validation of the bioanalytical method are presented. For human plasma samples a lower limit of quantification (LLOQ) of 5.00 pg/ml (=4.77 pmol/l) was achieved. The inter-assay precision was less than 11% and bias was within +/-4% over the whole validated range of 5.00-860 pg/ml.
由于这类化合物通常具有较高的效力，因此肽类药物给药后的血浆浓度往往较低。在这项工作中，我们提出了一种基于偶联柱液相色谱-串联质谱（LC-MS/MS）的生物分析方法，用于定量测定正在临床开发中的肽类药物 FE 202158。取 0.5ml 人血浆，用弱阳离子交换剂固相萃取。溶剂蒸发至干后，将复溶的样品注入偶联柱液相色谱系统。最初的氰基柱从初始柱上切下，被捕获在 C(4)柱上，然后注入微径 C(18)柱。采用配备 TurboIonSpray 接口的三重四极杆质谱仪，以正离子模式进行检测。本文介绍了该系统的设计，并讨论了与传统系统相比，该系统在灵敏度和选择性方面的提高。本文呈现了生物分析方法验证的数据。对于人血浆样品，可达到 5.00pg/ml（=4.77pmol/l）的定量下限（LLOQ）。在整个验证的 5.00-860pg/ml 范围内，批间精密度小于 11%，偏差在 +/-4%范围内。
