Simultaneous determination of tryptophan, kynurenine, kynurenic acid and two monoamines in rat plasma by HPLC-ECD/DAD.
J Pharm Biomed Anal
; 158: 8-14, 2018 Sep 05.
Article
en En
| MEDLINE
| ID: mdl-29843007
ABSTRACT
A high-performance liquid chromatography method with a diode array and an electrochemical detection (HPLC-ECD/DAD) was developed to determine the levels of tryptophan (TRP), kynurenine (KYN), kynurenic acid (KYA), 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in rat plasma. The prepared samples were separated on a BDS column (4.6â¯mmâ¯×â¯250â¯mm, 5â¯mm) with column oven temperature of 25⯰C. The mobile phase consisted of 5% acetonitrile and a buffer solution, which contained 25â¯mmol/L sodium acetate and 0.01â¯mmol/L EDTA, adjusting pH to 4.5 with acetic acid, and it was pumped at a flow-rate of 1.0â¯mL/min. KYN and KYA were measured by a variable wavelength detector at wavelengths 360â¯nm and 333â¯nm respectively, TRP and vanillic acid (as IS) both were measured at 280â¯nm. Determination of 5-HT and 5-HIAA was accomplished at the electrochemical working potential of 700â¯mV. Total run time was 14â¯min. Several parameters of the developed method were validated including linearity, accuracy precision, and stability. The results showed the established method had good LOD and separation for all of the five compounds and IS in the biological matrix. The method is simple, fast, economical and accurate. The analytical method and the results could provide a reference for the clinical and scientific research of depression.
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Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Triptófano
/
Técnicas Electroquímicas
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Aminoácidos Esenciales
Tipo de estudio:
Diagnostic_studies
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Etiology_studies
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Prognostic_studies
Límite:
Animals
Idioma:
En
Año:
2018
Tipo del documento:
Article