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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__________
#1
Title: Amino Acids in Infant Formula and Adult/Pediatric Formulas Ultra High Performance Liquid
Chromatography
Author:
TUV-SUD
Summary of Method:
Three separate digestions (acid hydrolysis, oxidative acid hydrolysis (Cys, Met), and
alkaline hydrolysis (Trp) for total amino acid analysis by dervitization with 6-aminoquinolyl-N-
hydrolyysuccinimidyl carbamate (AQC) reagent (except Trp). Dervitized amino acids are separated by
UHPLC and detected by DAD and FL (Trp) using Norvaline as internal standard.
Method Scope/Applicability:
Total amino acid analysis in infant, adult, and/or pediatric formula (powders,
RTF liquids and liquid concentrates) according to applicability statement however validation data only
shown for NIST 1849a.
General comments about the method:
Overall data from NIST SRM 1849a looks very promising. Separate digestions to prevent degradation of
Cys, Met, and Trp. Phenol addition in acid hydrolysis to prevent tyrosine degregation during hydrolysis.
Norvaline IS and is historically robust for AA analysis methods. UHPLC separation resulting in shorter
runtimes and high throughput sample preparation and analysis. Method also captures Taurine.
Method Clarity:
Phenol is added as a “speck” (approximately 10 mg)
Method Safety Concerns:
Acids and bases utilized in hydrolysis—exercise typical laboratory safety precautions.
Pros/Strengths:
•
Utilizes internal standard (Norvaline)
•
Addition of phenol to protect Tyr in acid hydrolysis
•
Fast UHPLC separation
•
Separate digestions to prevent degradation of Cys, Met, and Trp
Amino-03 Review Forms