High-performance liquid chromatography with fluorescence detection for quantitation of tryptophan and tyrosine in a shrimp waste protein concentrate

被引:39
作者
Sanchez-Machado, D. I. [1 ]
Chavira-Willys, B. [1 ]
Lopez-Cervantes, J. [1 ]
机构
[1] Inst Tecnol Sonora, Dept Biotecnol & Ciencias Alimentarias, Sonora, Mexico
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2008年 / 863卷 / 01期
关键词
tryptophan; tyrosine; high-performance liquid chromatography (HPLC); protein concentrate; shrimp waste; fluorescence detection;
D O I
10.1016/j.jchromb.2008.01.011
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A new, simple, and reproducible isocratic high-performance liquid chromatography (HPLC) method has been developed for the determination of free and total tyrosine and tryptophan in a protein concentrate. To determine total amino acids, the method involves alkaline hydrolysis of the proteins with sodium hydroxide at 120 degrees C for 4h in the absence of air. Best results were achieved with a SS Exil ODS column 5 mu m (25 cm x 0.46 cm i.d.), with an eluent of methanol: 40 mM sodium acetate buffer (adjusted to pH 4.5 with acetic acid; 20:80, v/v), a flow rate of 0.80 mL/min at 26 degrees C, and with programmable fluorescence detection. Under optimum conditions excellent linearity was obtained, and the overall recovery was 90.5, and 95.9% for total tryptophan and tyrosine, respectively. The precision results showed that the relative standard deviation of the repeatability and reproducibility were <= 4.78 and <= 4.65, respectively. This method was used to quantify the cited analytes in the protein concentrate obtained during the lactic acid fermentation of shrimp waste. (C) 2008 Elsevier B.V. All rights reserved.
引用
收藏
页码:88 / 93
页数:6
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