Determination of Amino Acids in Peptide Nutrition Powder by RP-HPLC with Pre-column Derivatization
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Abstract:
The method for determining the type and content of free amino acids in the peptide nutrition powder was established by using the column derivatization reverse high performance liquid chromatography (RP-HPLC) for the first time. The optimal mobile phase A were determined as sodium acetate - triethylamine buffer (pH 7.20 ± 0.05) with tetrahydrofuran (0.5%) to adjust the peak type. And the best mobile phase B was sodium acetate buffer (pH 7.20 + 0.05) - acetonitrile-methanol (20:40:40). O-phthalaldehyde (OPA) and Fluorene methyl chloroformate (FOMC-Cl) were used as the pre-column derivatization reagent. Separation was performed on an Agilent Hypersil ODS column (4.0 mm×250 mm, 5μm) with column temperature of 40 ℃. The detection wavelength was set at 338 nm /262 nm. 17 kinds of amino acids, including 8 essential amino acids, were determined by using the former method. The results showed that all the 17 amino acids were in good liner relationship. And the average recovery rates of the hydrolyzed amino acids in peptide nutrition powder of Perfect Brand ranged from 85% to 115% (RSD 1.15%~5.23%, n=6). At least 15 kinds of amino acids were detected in the powder. The results also indicated that the method was accurate, sensitive and simple, and proved to be suitable for the quantification of each amino acid in peptide nutrition powder of Perfect Brand.
