Free Physiologic Amino Acids MRM Method

Measured Analytes

  1. Phenylalanine
  2. Isoleucine
  3. Leucine
  4. Proline
  5. Valine
  6. Methionine
  7. Tyrosine
  8. Alanine
  9. Threonine
  10. Glutamate
  11. Serine
  12. Citrulline
  13. Histidine
  14. Glutamine
  15. Lysine
  16. Ornithine
  17. Arginine


Liquid chromatography tandem mass spectrometry (LC-MS) was carried out with a Shimadzu Prominence HPLC system and SCIEX 4000 mass spectrometer. An aliquot of 10 µl of each sample was injected onto an Normal Phase Intrada Amino Acid column 50 × 3 mm; 3 μm at 35oC for gradient separation. Mobile phases were: A) ddH2O with 100 mM ammonium formate; and B) Acetonitrile:water:formic acid (v:v:v = 95:5:0.3). Gradient schedule is the following: 0–3.0 min, 92%–88% Solution B; 3.0–6.4 min, 88%–70% Solution B; 6.4–6.5 min, 70%–0% Solution B; 6.5–10 min, 0% Solution B; 10–10.1 min, 0–92% Solution B; 10.1–13 min, 92% Solution B, with a total run time of 13 min. The flow rate is 600µL/min.

Arginine/Ornithine/Citruline MRM Chromatography

Extraction Protocols:

Extraction protocols for sample processing are dependent on the matrices involved. Please see extraction methods section for more specific instructions.


Method derived from the following

[1] Liu, Z., Tu, M.-J., Zhang, C., Jilek, J. L., Zhang, Q.-Y., & Yu, A.-M. (2019).
A reliable LC-MS/MS method for the quantification of natural amino acids in mouse plasma: Method validation and application to a study on amino acid dynamics during hepatocellular carcinoma progression.
Journal of Chromatography B, 1124, 72-81. doi:10.1016/j.jchromb.2019.05.039


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