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通过熔滴电喷雾电离质谱法消除蛋白质分析中TRIS缓冲液和SDS的干扰。

Eliminating the interferences from TRIS buffer and SDS in protein analysis by fused-droplet electrospray ionization mass spectrometry.

作者信息

Shieh I-Fan, Lee Chi-Yang, Shiea Jentaie

机构信息

Department of Chemistry, National Sun Yat-Sen University, Kaohsiung 804, Taiwan.

出版信息

J Proteome Res. 2005 Mar-Apr;4(2):606-12. doi: 10.1021/pr049765m.

Abstract

Multiply charged protein ions were detected from the solutions containing a high concentration of tris(hydroxymethyl) aminomethane buffer (TRIS) and sodium dodecyl sulfate (SDS) using fused-droplet electrospray ionization mass spectrometry (FD-ESI/MS). The sample aerosols were generated at ambient temperature with a pneumatic nebulizer commonly used to produce sample aerosols in an atmospheric pressure chemical ionization (APCI) source. The aerosols were carried by nitrogen gas to the tip of a capillary where charged methanol droplets had been continuously generated by electrospraying an acidic methanol solution. The neutral sample aerosols then fused with the charged methanol droplets and electrospray ionization proceeded from the newly formed fused droplets to generate multiply charged protein ions. Because of its low solubility in methanol, TRIS molecules (concentration as high as 1 M) were efficiently excluded from the newly formed droplets and the protein ion signals were detected and observed in the mass spectra. To remove the interferences from SDS, equal moles of positively charged cetyltrimethylammonium bromide (CTAB) was added into the SDS containing sample solution to form the dodecyl sulfate-cetyltrimethylammonium ion pair (DS-CTA). The DS-CTA ion pair has a low polarity and solubility in methanol and is excluded from the fused droplet. Protein ions were still detected from the solution containing 10(-2) M of SDS.

摘要

使用熔滴电喷雾电离质谱法(FD-ESI/MS)从含有高浓度三(羟甲基)氨基甲烷缓冲液(TRIS)和十二烷基硫酸钠(SDS)的溶液中检测到多电荷蛋白质离子。样品气溶胶在环境温度下由常用于在大气压化学电离(APCI)源中产生样品气溶胶的气动雾化器产生。气溶胶由氮气携带至毛细管尖端,在那里通过电喷雾酸性甲醇溶液持续产生带电的甲醇液滴。中性样品气溶胶随后与带电的甲醇液滴融合,电喷雾电离从新形成的融合液滴开始进行,以产生多电荷蛋白质离子。由于TRIS分子在甲醇中的溶解度低,其(浓度高达1 M)被有效地排除在新形成的液滴之外,并且在质谱图中检测并观察到蛋白质离子信号。为了消除SDS的干扰,将等摩尔的带正电荷的十六烷基三甲基溴化铵(CTAB)加入到含有SDS的样品溶液中,形成十二烷基硫酸根 - 十六烷基三甲基铵离子对(DS-CTA)。DS-CTA离子对在甲醇中的极性和溶解度低,并且被排除在融合液滴之外。从含有10^(-2) M SDS的溶液中仍然检测到蛋白质离子。

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