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通过电膜萃取从血浆中快速分离血管紧张素肽。

Rapid isolation of angiotensin peptides from plasma by electromembrane extraction.

作者信息

Balchen Marte, Halvorsen Trine Grønhaug, Reubsaet Léon, Pedersen-Bjergaard Stig

机构信息

Department of Pharmaceutical Chemistry, School of Pharmacy, University of Oslo, PO Box 1068 Blindern, 0316 Oslo, Norway.

出版信息

J Chromatogr A. 2009 Oct 9;1216(41):6900-5. doi: 10.1016/j.chroma.2009.08.037. Epub 2009 Aug 21.

Abstract

The present study has for the first time demonstrated the isolation of peptides from human plasma by electromembrane extraction (EME). Angiotensin 1, angiotensin 2, and angiotensin 3 migrated from 500 microL of diluted plasma, through a thin layer of 1-octanol and 8% di-(2-ethylhexyl) phosphate immobilized as a supported liquid membrane (SLM) in the pores of a porous hollow fiber, and into a 25 microL aqueous acceptor solution present inside the lumen of the fiber. The driving force for the extraction was a 15 V potential difference applied across the SLM. After only 10 min of EME, the peptides were isolated from diluted plasma (pH 3) with extraction recoveries between 25 and 43%. After optimization, the extraction system was evaluated using spiked plasma samples of angiotensin 2. The evaluation was performed by liquid chromatography electrospray mass spectrometry, showing linearity of angiotensin 2 in the range 2.5-125.0 ng/mL (r(2)=0.989), and repeatability (RSD) between 5.6 and 11.6% (n=6). The results demonstrate the possibility of isolating angiotensin peptides from plasma in only 10 min, using electromembrane extraction. The experimental findings are therefore promising with regard to future peptide extractions.

摘要

本研究首次展示了通过电膜萃取(EME)从人血浆中分离肽类物质的方法。血管紧张素1、血管紧张素2和血管紧张素3从500微升稀释血浆中迁移出来,穿过一层薄薄的以1-辛醇和8%二(2-乙基己基)磷酸固定在多孔中空纤维孔中的支撑液膜(SLM),进入纤维内腔中存在的25微升水相接受液中。萃取的驱动力是施加在支撑液膜上的15伏电位差。仅经过10分钟的电膜萃取,就从稀释血浆(pH值为3)中分离出了肽类物质,萃取回收率在25%至43%之间。优化后,使用血管紧张素2的加标血浆样本对萃取系统进行了评估。通过液相色谱电喷雾质谱法进行评估,结果显示血管紧张素2在2.5至125.0纳克/毫升范围内呈线性(r(2)=0.989),重复性(相对标准偏差)在5.6%至11.6%之间(n=6)。结果表明,利用电膜萃取仅需10分钟就能从血浆中分离出血管紧张素肽类物质。因此,这些实验结果对于未来的肽类萃取很有前景。

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