期刊
出版社
ELSEVIER
DOI: 10.1016/j.jchromb.2008.01.011
关键词
tryptophan; tyrosine; high-performance liquid chromatography (HPLC); protein concentrate; shrimp waste; fluorescence detection
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.
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