LIQUID-CHROMATOGRAPHIC DETERMINATION OF AMINO-ACIDS AFTER GAS-PHASE HYDROLYSIS AND DERIVATIZATION WITH (DIMETHYLAMINO)AZOBENZENESULFONYL CHLORIDE
LIQUID-CHROMATOGRAPHIC DETERMINATION OF AMINO-ACIDS AFTER GAS-PHASE HYDROLYSIS AND DERIVATIZATION WITH (DIMETHYLAMINO)AZOBENZENESULFONYL CHLORIDE
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DOI：
10.1021/ac00125a006
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发表时间：
1986-10-01
影响因子：
7.4
通讯作者：
CHANG, JY
中科院分区：
文献类型：
--
作者：
KNECHT, R;CHANG, JY
空白样品中氨基酸背景水平是进行高灵敏度氨基酸分析的限制因素之一。采用（二甲氨基）偶氮磺酰氯（DABS-Cl）柱前标记/高效液相色谱法，系统地研究了水解和衍生化过程中潜在的污染物来源。数据表明，氨基酸污染物最可能的来源是水溶液中的6N HCl。气相酸水解可使正常液相水解产生的本底降低到25%左右。一个典型的空白样品，经过气相水解，平均产生0.5- 3pmol的Asp， Glu， Ser和Gly。这种低水平的背景与改进的DABS-Cl方法一起允许对低纳克量的蛋白质水解物进行常规和可靠的氨基酸分析。
The level of amino acid background in the blank sample is one of the limiting factors for high sensitivity amino acid analysis. By use of the (dimethylamino)azobenzenesulfonyl chloride (DABS-Cl) precolumn labeling/HPLC method, the potential sources of contaminants during hydrolysis and derivatization were systematically investigated. The data indicate that the most likely source of amino acid contaminants is aqueous 6N HCl. Gas-phase acid hydrolysis can reduce the background generated by normal liquid-phase hydrolysis to about 25%. A typical blank sample, after gas-phase hydrolysis, yields an average of 0.5-3 pmol of Asp, Glu, Ser, and Gly. This low level of background together with the improved DABS-Cl method permits routine and reliable amino acid analysis of low nanogram quantities of protein hydrolysates.