Proteomics > Which service should I request? > Amino Acid Analysis > Amino acid - Free amino acids
The workflow consists of four major steps
Amino acid - Free amino acids
Table of contents
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
- Protein precipitation: proteins are removed by 10% 5-sulfosalicylic acid (SSA)
- Labeling: reaction with Accq Tag Ultra (aminoquinolyl-N-hydroxysuccinimidyl carbamate). The reaction is shown below.
- LC-QDA analysis: Cortex UPLC reverse-phase separation of the amino acid derivatives
- Data analysis: Data are processed by TargetLynx™ of MassLynx™ Software (Waters)(MassLynx)
- Calibration curve establishment:
- 2.5 uM to 200 uM per amino acid Standard H plus Gln, Asn and Trp
- or based on the free amino acids of interest plus isotope-labeled amino acid mixture
- amino acid quantification
- Calibration curve establishment:
List of free amino acids
The list of free amino acids that can be monitored:Amino acids | Abbreviation | MW |
γ-amino-n-butyric acid | GABA | 103.1 |
L-Anserine | Ans | 240.3 |
L-Arginine | Arg | 174.2 |
L-Carnosine | car | 226.2 |
Creatinine | 113.1 | |
Ethanolamine | EA | 61.08 |
L-Histidine | His | 155.2 |
L-Homocystine | (Hcys)2 | 268.3 |
DL-hydroxylysine | Hyl | 162.2 |
L-lysine | Lys | 146.2 |
1-Methyl-L-histidine | 1MeHis | 169.2 |
3-Methyl-L-histidine | 3MeHis | 169.2 |
L-Ornithine | Orn | 132.2 |
L-Tryptophan | Trp | 204.2 |
β-alanine | bAla | 89.09 |
L-alanine | Ala | 89.09 |
L-α-aminoadipic acid | Aad | 161.2 |
L-α-amino-n-butyric acid | Abu | 103.1 |
L-β-amino-isobutyric acid | bAib | 103.1 |
L-Asparagine | Asn | 132.1 |
L-Asparticacid | Asp | 133.1 |
L-Citrulline | Cit | 175.2 |
L-Glutamic acid | Glu | 147.1 |
Glycine | Gly | 75.07 |
Hydroxy-L-Proline | Hyp | 131.1 |
L-Isoleucine | Ile | 131.2 |
L-Leucine | Leu | 131.2 |
L-Methionine | Met | 149.2 |
L-Phenylalanine | Phe | 165.2 |
O-phospo-L-Serine | pSer | 185.1 |
O-phospoethanolamine | PEA | 141.4 |
L-Proline | Pro | 115.4 |
L-Sarcosine | Sar | 89.09 |
L-Serine | Ser | 105.1 |
Taurine | Tau | 125.1 |
L-Threonine | Thr | 119.1 |
L-Tyrosine | Tyr | 181.2 |
L-Valine | Val | 117.2 |
L-Glutamine | Gln | 146.1 |
L-allo-Isoleucine | alle | 131.17 |
L-Homocitrulline | Hci | 189.21 |
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.