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Simultaneous analysis of kynurenine and tryptophan in human plasma by capillary electrophoresis with UV detection

Zinellu, Angelo and Sotgia, Salvatore and Deiana, Luca and Talanas, Giuseppe and Terrosu, Pierfranco and Carru, Ciriaco (2012) Simultaneous analysis of kynurenine and tryptophan in human plasma by capillary electrophoresis with UV detection. Talanta, Vol. 35 (9), p. 1146-1151. eISSN 1873-3573. Article.

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DOI: 10.1002/jssc.201200021

Abstract

Plasma kynurenine (Kyn)/tryptophan ratio has been proposed as a useful marker for the monitoring activation of the cellular immune system. Here, we describe an easy capillary electrophoresis method with UV detection for the separation and detection of Kyn and tryptophan in human plasma using methltryptophan as internal standard. The plasma samples were simply treated with acentonitrile for the elimination of proteins, the supernatant was evaporated, and the dried sample was resuspended with water and directly injected on the capillary without sample derivatization procedures. The use of a run buffer composed by 100 mmol/L Bis-Tris propane at pH 2.15 allowed to baseline resolve the analytes within 9 min. Precision tests indicated a good repeatability of our method both for times (CV < 0.53%) and areas (CV < 2.8%). Moreover, a good reproducibility of intra-assay and interassay tests was obtained (CV < 3.9% and CV < 7.6%, respectively). The obtained limit of detections for Kyn and tryptophane, evaluated at 226 nm, were 0.15 and 0.40 mol/L, respectively. The method suitability was tested by measuring analyte levels both in healthy volunteers, acute myocardial infarction and chronic kidney disease patients.

Item Type:Article
ID Code:7766
Status:Published
Refereed:Yes
Uncontrolled Keywords:Capillary electrophoresis, kynurenine, L-tryptophan
Subjects:Area 05 - Scienze biologiche > BIO/12 Biochimica clinica e biologia molecolare clinica
Divisions:002 Altri enti e centri di ricerca del Nord Sardegna > Azienda ASL1, Sassari > Ospedale civile SS. Annunziata
002 Altri enti e centri di ricerca del Nord Sardegna > Porto Conte Ricerche, Alghero
002 Altri enti e centri di ricerca del Nord Sardegna > Azienda ASL1, Sassari
001 Università di Sassari > 01-a Nuovi Dipartimenti dal 2012 > Scienze Biomediche
Publisher:Elsevier
eISSN:1873-3573
Deposited On:12 Jul 2012 10:02

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