Determination of amino acids by capillary electrophoresis-electrospray ionization-mass spectrometry: An evaluation of different protein hydrolysis procedures

被引:48
作者
Colnaghi Simionato, Ana V. [1 ,2 ]
Moraes, Edgar Perin [1 ]
Carrilho, Emanuel [2 ]
Maggi Tavares, Marina Franco [1 ]
Kenndler, Ernst [3 ]
机构
[1] Univ Sao Paulo, Inst Quim, BR-05508900 Sao Paulo, Brazil
[2] Univ Sao Paulo, Inst Chem Sao Carlos, BR-13560 Sao Carlos, Brazil
[3] Univ Vienna, Inst Analyt Chem, A-1010 Vienna, Austria
关键词
amino acids; cation exchanger resin; capillary electrophoresis-electrospray ionization-mass spectrometry; hydrolysis methods;
D O I
10.1002/elps.200700629
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In this work, a CE equipment, online hyphenated to an IT MS analyzer by a linear sheath liquid interface promoting ESI, was used to develop a method for quantitative determination of amino acids. Under appropriate conditions (BGE composition, 0.8% HCOOH, 20% CH3OH; sheath liquid composition, 0.8% HCOOH, 60% methanol; V-ESI, +4.50 W), analytical curves of all amino acids from 3 to 80 mg/L were recorded presenting acceptable linearity (r > 0.99). LODs in the range of 16-172 mu mol/L were obtained. BSA, a model protein, was submitted to different hydrolysis procedures (classical acid and basic, and catalyzed by the H+ form of a cation exchanger resin) and its amino acid profiles determined. In general, the resin-mediated hydrolysis yields were overall similar or better than those obtained by classical acid or basic hydrolysis. The resulting experimental-to-theoretical BSA concentration ratios served as correction factors for the quantitation of amino acids in Brazil nut resin generated hydrolysates.
引用
收藏
页码:2051 / 2058
页数:8
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