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Proteomics > Which service should I request? > Amino Acid Analysis > Amino acid - Free amino acids

Amino acid - Free amino acids




General description

Obtain free amino acid in physiological fluid (i.e.: urine, plasma), tissues and a myriad of other biological samples.

Workflow


The workflow consists of four major steps
  1. Protein precipitation: proteins are removed by 10% 5-sulfosalicylic acid (SSA)
  2. Labeling: reaction with Accq Tag Ultra (aminoquinolyl-N-hydroxysuccinimidyl carbamate). The reaction is shown below.
    Image
  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)

List of free amino acids

The list of free amino acids that can be monitored:

Amino acidsAbbreviationMW
γ-amino-n-butyric acid GABA 103.1
L-AnserineAns240.3
L-ArginineArg174.2
L-Carnosinecar226.2
Creatinine 113.1
EthanolamineEA61.08
L-HistidineHis155.2
L-Homocystine(Hcys)2268.3
DL-hydroxylysineHyl162.2
L-lysineLys146.2
1-Methyl-L-histidine1MeHis169.2
3-Methyl-L-histidine3MeHis169.2
L-OrnithineOrn132.2
L-TryptophanTrp204.2
β-alaninebAla89.09
L-alanineAla89.09
L-α-aminoadipic acidAad161.2
L-α-amino-n-butyric acidAbu103.1
L-β-amino-isobutyric acidbAib103.1
L-AsparagineAsn132.1
L-AsparticacidAsp133.1
L-CitrullineCit175.2
L-Glutamic acidGlu147.1
GlycineGly75.07
Hydroxy-L-ProlineHyp131.1
L-IsoleucineIle131.2
L-LeucineLeu131.2
L-MethionineMet149.2
L-PhenylalaninePhe165.2
O-phospo-L-SerinepSer185.1
O-phospoethanolaminePEA141.4
L-ProlinePro115.4
L-SarcosineSar89.09
L-SerineSer105.1
TaurineTau125.1
L-ThreonineThr119.1
L-TyrosineTyr181.2
L-ValineVal117.2
L-GlutamineGln146.1
L-allo-Isoleucinealle131.17
L-HomocitrullineHci189.21


Requirements and considerations: 

  • optimal volume: > 50 uL
  • volume indicated are for urine and plasma samples. They may be significantly different from different species or tissue extracts.
  • SSA precipitation has to be performed freshly before the labeling. Please don't perform SSA precipitation for samples.


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