Abstract
Background: Plasma amino acids are usually analyzed by ion-exchange chromatography (IEC), a reproducible but time consuming method. Here, we test whether plasma amino acids can be analyzed using reverse-phase high performance liquid chromatography (HPLC). Methods: Filtered plasma, with S-carboxymethyl-l-cysteine as the internal standard, was derivatized and analyzed by an Agilent 1100 HPLC system. Primary amino acids were derivatized with o-phthalaldehyde 3-mercaptopropionic acid (OPA) and detected by a diode array detector. Secondary amino acids were derivatized with 9-fluorenylmethyl chloroformate (FMOC) and detected fluorometrically. Chromatographic separation is achieved by two gradient elutions (two injections per sample), starting at different pHs, on a reverse phase Agilent Zorbax Eclipse C18 column AAA (4.6×150 mm). Results: The HPLC method evaluated correlated well with IEC (0.89≤r≤1.00) with linearity up to 2500 μmol/l. The between- and within-run CVs were
Original language | English (US) |
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Pages (from-to) | 83-90 |
Number of pages | 8 |
Journal | Clinica Chimica Acta |
Volume | 354 |
Issue number | 1-2 |
DOIs | |
State | Published - Jan 1 2005 |
Externally published | Yes |
Bibliographical note
Generated from Scopus record by KAUST IRTS on 2023-09-20ASJC Scopus subject areas
- Biochemistry
- Clinical Biochemistry
- Biochemistry, medical