采用高效液相色谱-电化学检测法测定皮克级的脂氧素A4和脂氧素B4。

Determination of picogram amounts of lipoxin A4 and lipoxin B4 by high-performance liquid chromatography with electrochemical detection.

作者信息

Herrmann T, Steinhilber D, Knospe J, Roth H J

机构信息

Pharmaceutical Institute, University of Tübingen, F.R.G.

出版信息

J Chromatogr. 1988 Jul 15;428(2):237-45. doi: 10.1016/s0378-4347(00)83914-6.

DOI:10.1016/s0378-4347(00)83914-6

PMID:3215927

Abstract

A new method for the determination of lipoxins with electrochemical detection after high-performance liquid chromatography is described. The half-wave potentials of lipoxin A4 and lipoxin B4 at a glassy carbon electrode and a mobile phase of methanol-water (65:35 v/v) and 1 mM trifluoroacetic acid was found to be +1.14 V versus an Ag/AgCl reference electrode. The use of trifluoroacetic acid instead of sulphuric acid and lithium perchlorate led to a background current of 6-8 nA at +1.20 V. The detection limits for both lipoxins, based on a signal-to-noise ratio of 3:1 were found to be 5-10 pg (15-30 fmol). The new method was applied to an extract of human polymorphonuclear granulocytes, preincubated with 15-hydroxyeicosatetraenoic acid and stimulated with Ca2+ ionophore A23187.

摘要

描述了一种在高效液相色谱后用电化学检测法测定脂氧素的新方法。发现脂氧素A4和脂氧素B4在玻碳电极上的半波电位以及甲醇 - 水（65:35 v/v）和1 mM三氟乙酸的流动相相对于Ag/AgCl参比电极的半波电位为 +1.14 V。使用三氟乙酸代替硫酸和高氯酸，在 +1.20 V时产生的背景电流为6 - 8 nA。基于3:1的信噪比，两种脂氧素的检测限均为5 - 10 pg（15 - 30 fmol）。该新方法应用于预先用15 - 羟基二十碳四烯酸孵育并用Ca2+离子载体A23187刺激的人多形核粒细胞提取物。