采用荧光检测的毛细管电泳法测定唾液中的氨基酸。

Determination of amino acids in saliva using capillary electrophoresis with fluorimetric detection.

作者信息

Pobozy Ewa, Czarkowska Wioletta, Trojanowicz Marek

机构信息

Departament of Chemistry, Warsaw University, Pasteura 1, 02-093 Warsaw, Poland.

出版信息

J Biochem Biophys Methods. 2006 Apr 30;67(1):37-47. doi: 10.1016/j.jbbm.2006.01.001. Epub 2006 Feb 3.

DOI:10.1016/j.jbbm.2006.01.001

PMID:16472865

Abstract

In the present study a sensitive method for the quantification of main free amino acids in saliva using capillary electrophoresis with laser induced fluorescence detection was developed. As background electrolyte 20 mM borate buffer pH 9.5 was used. Amino acids were separated after derivatization with fluorescein isothiocyanate (FITC) and the conditions for derivatization were optimized. The main amino acids occurring in saliva (Pro, Ser, Gly and Glu) were separated in less than 7 min. The parameters of validation such as linearity of response, precision and detection limits were determined. The detection limits were obtained in the range from 0.1 to 2.4 nM. The developed method was employed for determination of amino acids in real saliva samples.

摘要

在本研究中，开发了一种使用毛细管电泳结合激光诱导荧光检测法定量唾液中主要游离氨基酸的灵敏方法。使用pH 9.5的20 mM硼酸盐缓冲液作为背景电解质。氨基酸经异硫氰酸荧光素（FITC）衍生化后进行分离，并对衍生化条件进行了优化。唾液中主要的氨基酸（脯氨酸、丝氨酸、甘氨酸和谷氨酸）在不到7分钟内即可分离。测定了响应线性、精密度和检测限等验证参数。检测限在0.1至2.4 nM范围内。所开发的方法用于实际唾液样品中氨基酸的测定。