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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。

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Determination of picogram amounts of lipoxin A4 and lipoxin B4 by high-performance liquid chromatography with electrochemical detection.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。
J Chromatogr. 1988 Jul 15;428(2):237-45. doi: 10.1016/s0378-4347(00)83914-6.
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Determination of picomole amounts of lipoxins C4, D4 and E4 by high-performance liquid chromatography with electrochemical detection.采用高效液相色谱-电化学检测法测定皮摩尔量的脂氧素C4、D4和E4。
J Chromatogr. 1991 Oct 25;585(1):53-6. doi: 10.1016/0021-9673(91)85056-l.
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Identification of a novel 7-cis-11-trans-lipoxin A4 generated by human neutrophils: total synthesis, spasmogenic activities and comparison with other geometric isomers of lipoxins A4 and B4.人中性粒细胞产生的新型7-顺式-11-反式-脂氧素A4的鉴定:全合成、致痉活性及其与脂氧素A4和B4其他几何异构体的比较
Biochim Biophys Acta. 1989 May 15;1003(1):44-53. doi: 10.1016/0005-2760(89)90096-9.
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FEBS Lett. 1989 Sep 11;255(1):143-8. doi: 10.1016/0014-5793(89)81078-6.
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Transcellular conversion of endogenous arachidonic acid to lipoxins in mixed human platelet-granulocyte suspensions.在人血小板-粒细胞混合悬液中内源性花生四烯酸向脂氧素的跨细胞转化。
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Lipoxin A4 and lipoxin B4 stimulate the release but not the oxygenation of arachidonic acid in human neutrophils: dissociation between lipid remodeling and adhesion.脂氧素A4和脂氧素B4刺激人中性粒细胞中花生四烯酸的释放,但不影响其氧化:脂质重塑与黏附之间的分离。
J Cell Physiol. 1990 Jun;143(3):512-23. doi: 10.1002/jcp.1041430316.

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Human alveolar macrophages have 15-lipoxygenase and generate 15(S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid and lipoxins.
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Formation of lipoxins and leukotrienes during receptor-mediated interactions of human platelets and recombinant human granulocyte/macrophage colony-stimulating factor-primed neutrophils.人血小板与重组人粒细胞/巨噬细胞集落刺激因子预激活的中性粒细胞在受体介导相互作用过程中脂氧素和白三烯的形成。
J Exp Med. 1990 Nov 1;172(5):1451-7. doi: 10.1084/jem.172.5.1451.