黄金蝉若虫体蛋白水解液的荧光标记及电喷雾质谱鉴定

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	黄金蝉若虫体蛋白水解液的荧光标记及电喷雾质谱鉴定
	李肖燕 ; 尤进茂 ; 孙志伟 ; 付艳艳 ; 秦雪芹 ; 丁明州 ; 李玉林
	2010
发表期刊	分析化学 ; 李肖燕;尤进茂;孙志伟;付艳艳;秦雪芹;丁明州;李玉林.黄金蝉若虫体蛋白水解液的荧光标记及电喷雾质谱鉴定,分析化学,2010,38(7):917-923
摘要	摘　要　采用22[ 22(7H2二苯并[ a, g]咔唑2乙氧基) 2乙基]氯甲酸酯(DBCEC2Cl)作为柱前衍生试剂, 于室温下,在硼酸钠缓冲液( pH 9. 0)中,金蝉若虫体蛋白水解液中氨基酸的完全衍生化在5 min即可完成。在 Hypersil BDS C18色谱柱(200 mm ×4. 6 mm, 5μm) 上, 采用梯度洗脱实现了各衍生物的完全基线分离。最佳荧光检测为λex /λem = 300 /395 nm。在线串联的ESI/MS正离子质谱数据显示: 所有衍生物均给出 m / z 33815, 294. 5的特征碎片离子峰。荧光条件下的线性回归系数大于0. 9992; 检出限为2. 6～24 fmol。本方法灵敏、可靠、重现性好。对实际黄金蝉若虫体蛋白水解液中氨基酸的测定,结果满意。 Abstract　By using 22[ 22(7H2dibenzo[ a, g ] carbazole2ethoxy) 2ethyl chloroformate (DBCEC2Cl) as a sensi2 tive p re2column derivatization reagent, a simp le method for evaluating the chemical compositions of p rotein amino acids from the hydrolyzed cicada samp le has been developed. Studies on derivatization conditions indicated that the labeling reaction p roceeded rap idly and smoothly in the p resence of a base catalyst (pH 910) in acetonitrile to give the corresponding sensitively fluorescent derivatives with an excitation maxi2 mum atλex 300 nm and an emission maximum atλem 395 nm. The separation of amino acid derivatives was carried out on a reversed phase Hypersil BDS2C18 ( 200 mm ×4. 6 mm ×5 μm) column in combination of a gradient elution. The comp lete baseline resolution for 20 amino acid derivatives was achieved within 65 min. Identification of amino acid derivatives was carried out by the online post2column ion trap /mass spectrometry with electrosp ray ion source ( ESI source) in positive ion mode. Excellent linear response was observed, with coefficients of > 0. 9992. Fluorescent detection limits calculated from 25 pmol injection, at a signal2to2noise ratio of 3, were 2. 6 - 24 fmol. The established method for the determination of amino acids from cicada sample was satisfactory.
文献类型	期刊论文
条目标识符	http://210.75.249.4/handle/363003/36239
专题	中国科学院西北高原生物研究所
推荐引用方式 GB/T 7714	李肖燕,尤进茂,孙志伟,等. 黄金蝉若虫体蛋白水解液的荧光标记及电喷雾质谱鉴定[J]. 分析化学, 李肖燕;尤进茂;孙志伟;付艳艳;秦雪芹;丁明州;李玉林.黄金蝉若虫体蛋白水解液的荧光标记及电喷雾质谱鉴定,分析化学,2010,38(7):917-923,2010.
APA	李肖燕.,尤进茂.,孙志伟.,付艳艳.,秦雪芹.,...&李玉林.(2010).黄金蝉若虫体蛋白水解液的荧光标记及电喷雾质谱鉴定.分析化学.
MLA	李肖燕,et al."黄金蝉若虫体蛋白水解液的荧光标记及电喷雾质谱鉴定".分析化学 (2010).