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Amino acid - Composition / Quantification

General description

There are many different methodologies to hydrolyze protein/peptide into amino acids. The main protocol used at FGCZ is using acid hydrolysis. This procedure allows to obtain higher efficiency of acid hydrolysis but the side chains of Gln and Asn will convert to Glu and Asp. In addition, tryptophan would be destroyed under this condition. Therefore, only 17 amino acids will be reported in this analysis.


The workflow consists of four major steps:
  1. Acid hydrolysis: proteins/peptides are hydrolyzed together with isotope labelled amino acids under 6M HCl with 0.1% w/v phenol stream at 110°C for 24 hours
  2. Labeling: reaction with Accq Tag Ultra (aminoquinolyl-N-hydroxysuccinimidyl carbamate). The reaction is shown below:
  3. LC-QDA analysis: Cortex UPLC reverse-phase separation of the amino acid derivatives
  4. Data analysis: Data are processed by TargetLynx™ of MassLynx™ Software (Waters)(MassLynx)

Requirements and considerations

  • volume: > 5 uL
  • minimal amount: 1-5 ug; optimal amount: 10 ug
  • dissolve samples in volatile solvents or free of non-volatile buffers, salts, detergents, and primary amine

Created by paolo. Last Modification: 2023-02-14 14:32 by paolo.