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FOR EXPERT REVIEW PANEL USE ONLY
NOT FOR DISTRIBUTION
1
Expert Review Panel for Infant Formula and Adult Nutrition
Evaluation of Method: Amino -03
#2
Title:
Amino Acids in Infant Formula and Adult/Pediatric Formulas
Ultra High Performance Liquid Chromatography
Author:
TUV-SUD
Summary of Method:
Method uses 3 different protein hydrolysis procedures:
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acid hydrolysis for Histidine, Taurine, Serine, Arginine, Glycine, Aspartic acid (sum of Aspartic acid
and Asparagine), Glutamic acid (sum of Glutamic acidand Glutamine), Threonine, Alanine, Proline,
Lysine, Tyrosine,Valine, Isoleucine, Leucine, Phenyl alanine.
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Oxidative hydrolysis with performic acid for Cysteine, Cystine and Methionine
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Alkaline hydrolysis for Tryptophan
Resulting amino acids (except Tryptophan) are derivitized with 6-aminoquinolyl-N-hydroxysuccinimidyl
carbamate (AQQ-Tag derivatization reagent from Waters) reagent and separated using UHPLC system.
(AA The detection of the derivatives is done using UV/Vis detector, Tryptophan is detected using
Fluorescence detector.
Norvaline is used as Internal Standard
Method Scope/Applicability:
Applicable for the determination of amino acids (including Taurine) in all
forms of infant, adult, and/or pediatric formula (powders, ready to feed liquids and liquid concentrates).
General comments about the method:
3 separate sample preparation procedures and HPLC methods are required to analyze all the amino acids
so essentially these are 3 separate methods. The hydrolysis procedures used are well-established and are
part of AOAC and ISO methods.
Experimental part is clearly written and should be is easy to follow.
The only data available are for SRM 1849a. No SPIFAN matrices were analyzed.
Amino-03 Review Forms