Liquid chromatography method for simultaneous analysis of amino acids and biogenic amines in biological fluids with simultaneous gradient of pH and acetonitrile

Liquid chromatography method for simultaneous analysis of amino acids and biogenic amines in biological fluids with simultaneous gradient of pH and acetonitrile

Liquid chromatography method for simultaneous analysis of amino acids and biogenic amines in biological fluids with simultaneous gradient of pH and acetonitrile

Application Number
122400
Method
HPLC
Results

Legend:
Chromatograms: (a) standard sample contains 130 pmol of each amino acids and 5 pmol of each polyamines, catecholamines and metanephrines per 20 µL injection (b) plasma sample His - histidine Arg - arginine Ser - serine Gln - glutamine Asn - asparagine Asp - aspartic acid Glu - glutamic acid Thr - threonine Ala - alanine GABA - γ-aminobutyric acid Pro - proline Met - methionine Val - valine Phe - phenylalanine Ile - isoleucine Leu - leucine 2-AH - 2-aminoheptanoic acid (IS) Orn - ornithine Lys - lysine Tyr - tyrosine Put - putrescine NME - normetaneprine Cad - cadaverine ME - metanephrine NE - norepinephrine Spd - spermidine E - epinephrine D - dopamine Spm - spermine

Substances
(His) histidine; (Arg) arginine; (Ser) serine; (Gln) glutamine; (Asn) asparagine; (Asp) aspartic acid; (Glu) glutamic acid; (Thr) threonine; (Ala) alanine; (GABA) γ-aminobutyric acid; (Pro) proline; (Met) methionine; (Val) valine; (Phe) phenylalanine; (Ile) isoleucine; (Leu) leucine; 2-aminoheptanoic acid; (Orn) ornithine; (Lys) lysine; (Tyr) tyrosine; (Put) putrescine; normetaneprine; (Cad) cadaverine; metanephrine; norepinephrine; (Spd) spermidine; epinephrine; dopamine; (Spm) spermine
Product(s)
PhaseREFWebshop

NUCLEODUR C18 ec

760001.40

Matrix
plasma
Sample(s)

For extraction, protein precipitation and derivatization with Fmoc-Cl, please see cited literature.

Conditions
  • Eluent A: 20 mmol/L ammonium acetate - acetonitrile (95:5, v/v) + 1 mmol/L dibutylamine, pH 2.1 (adjusted with 35 % perchloric acid)
  • Eluent B: 20 mmol/L ammonium acetate - acetonitrile (90:10, v/v) + 5 mmol/L dibutylamine, resulting pH 8.4
  • Gradient: 74 - 69 % A in 5 min, 69 - 63 % A in 10 min, 63 - 52 % A in 1 min, 52 - 44 % A in 5 min, 44-32 % A in 4 min, 32 - 14 % A in 2 min, 14 - 0 % A in 3 min, 0 % A for 6 min, 0 - 74 % A in 2 min, 74 % A for 5 min
  • Flow rate: 1.5 mL/min
  • Injection volume: 20 µL
Detection
Fluorescence, ex. 262 nm, em. 615 nm
Note
Author
Lozanov, V. et al.
Source
J. Chromatogr. B 860 (2007), 92 - 97
Disclaimer
Concerning questions to literature applications please contact the cited author/s. Chemicals mentioned in applications are not manufactured or sold by MACHEREY-NAGEL. Applications mentioned in MACHEREY-NAGEL literature or databases are provided for informational purposes only. MACHEREY-NAGEL does not warrant that all applications have been tested in MACHEREY-NAGEL laboratories using MACHEREY-NAGEL products. MACHEREY-NAGEL does not warrant the correctness of the applications.
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