高效液相色谱纯化的绵羊血浆中μ阿片受体激动剂酪氨酰-D-精氨酰-苯丙氨酰-赖氨酰胺（DALDA）的质谱定量分析

Mass spectrometric quantification of the mu opioid receptor agonist Tyr-D-Arg-Phe-Lys-NH2 (DALDA) in high-performance liquid chromatography-purified ovine plasma.

作者信息

Grigoriants O O, Tseng J L, Becklin R R, Desiderio D M

机构信息

The Charles B. Stout Neuroscience Mass Spectrometry Laboratory, University of Tennessee, Memphis 38163, USA.

出版信息

J Chromatogr B Biomed Sci Appl. 1997 Aug 1;695(2):287-98. doi: 10.1016/s0378-4347(97)00182-5.

DOI:10.1016/s0378-4347(97)00182-5

PMID:9300865

Abstract

The mu opioid receptor agonist Tyr-D-Arg-Phe-Lys-Amide (D-Arg2-Lys4-Dermorphin(1-4)amide=DALDA) was infused continuously for 2 h into sheep. The presence of DALDA in ovine plasma was determined by reversed-phase high-performance liquid chromatography (RP-HPLC) and mass spectrometry (MS) in plasma samples that were obtained at different times during and following that infusion. A stable isotope-incorporated internal standard, deuterated DALDA (d5-DALDA), was used for the MS quantification of DALDA via the protonated molecule ion, (M+H)+, of DALDA and of d5-DALDA. Time-course data (microg DALDA ml(-1) plasma vs. time) were obtained. Tandem MS (MS-MS) provided the product-ion spectrum of the (M+H)+ ion of DALDA in one of the samples to confirm the amino acid sequence of DALDA.

摘要

将μ阿片受体激动剂酪氨酰-D-精氨酰-苯丙氨酰-赖氨酰胺（D-Arg2-Lys4-德尔吗啡(1-4)酰胺 = DALDA）持续输注到绵羊体内2小时。通过反相高效液相色谱法（RP-HPLC）和质谱法（MS），在输注期间及输注后不同时间采集的血浆样本中测定绵羊血浆中DALDA的存在情况。一种稳定同位素掺入的内标，氘代DALDA（d5-DALDA），用于通过DALDA和d5-DALDA的质子化分子离子（M+H）+对DALDA进行质谱定量。获得了时间进程数据（微克DALDA/毫升血浆与时间的关系）。串联质谱（MS-MS）在其中一个样本中提供了DALDA的（M+H）+离子的产物离子谱，以确认DALDA的氨基酸序列。