Simultaneous analysis of kynurenine and tryptophan in human plasma by capillary electrophoresis with UV detection.
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 μmol/L, respectively. The method suitability was tested by measuring analyte levels both in healthy volunteers, acute myocardial infarction and chronic kidney disease patients.
Department of Biomedical Sciences, University of Sassari, Sassari, Italy. angelozinellu@libero.it
Angelo
A
Zinellu
Salvatore
S
Sotgia
Luca
L
Deiana
Giuseppe
G
Talanas
PierFranco
P
Terrosu
Ciriaco
C
Carru
D000368:Aged; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D006801:Humans; D007737:Kynurenine / Q000097:blood*; D008297:Male; D008875:Middle Aged; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D014364:Tryptophan / Q000097:blood*
D007737:Kynurenine; D014364:Tryptophan