Amino acid analysis is a considerable application applied in research, clinical facilities and industrial processes. A rapid and sensitive UHPLC method for the determination of amino acid concentrations and compositions in food samples has been worked out in this application note. Detection by UV and also fluorescence can be used to underline the variability of the presented method. Using a KNAUER Euorspher (former Bluespher) 100 x 2 mm ID column and a high speed gradient method on the KNAUER UHPLC system, a complex mixture of 17 derivatized amino acids could be separated in less than 8 minutes. Short Eurospher columns filled with small particles are the most suitable way for avoiding long equilibration and analysis times. The method described in this study uses AQC as precolumn derivatizing reagent. AQC is an excellent derivatization reagent for amino acid analysis by UHPLC caused by its fast and easy operability. The amino acid-AQC derivatives are substantially more stable than other commonly used reagents like orthophthaldialdehyde (OPA) or 9-fluorenyl-methoxycarbonyl chloride (Fmoc).
1 Asp, 2 Glu, 3 Ser, 4 Gly, 5 ?, 6 Lys, 7 His, 8 Thr, 9 Arg, 10 Ala, 11 AMQ, 12 Pro, 13 NH3, 14 Cys, 15 Tyr, 16 Val, 17 Met, 18 Ile, 19 Leu, 20 Lys 2, 21 Phe
Method | UHPLC |
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Mode | RP |
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Substances | Aspartic acid, Glutamic acid, Serine, Glycine, Lysine, Histidine, Threonine, Arginin, Alanine, Proline, Cystine, Tyrosine, Valine, Methionine, Isoleucine, Leucine, Lysine, Phenlyalanine |
Key words | AQC-derivatization, Amino Acids |
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