Thin‐layer chromatography—an image‐processing method for the determination of acidic catecholamine metabolites

• Published in: Journal of separation science Vol. 37; no. 19; pp. 2675 - 2681
• Main Authors: Casoni, Dorina, Sima, Ioana Anamaria, Sârbu, Costel
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley-VCH 01.10.2014; Blackwell Publishing Ltd; Wiley; Wiley Subscription Services, Inc
• Subjects: Acidic catecholamine metabolites; Biological and medical sciences; Catecholamine; Catecholamines - metabolism; Catecholamines - urine; Chromatography; Chromatography, Thin Layer - methods; detection limit; homovanillic acid; Human; Humans; Image processing systems; Image-processing analysis; Investigative techniques, diagnostic techniques (general aspects); Medical sciences; Metabolites; Miscellaneous. Technology; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Phase composition; Radical detection; Radicals; Spots; Thin-layer chromatography; Urine; Human; Urine; Validation; Biological fluid; Chemical analysis; DPPH; Healthy subject; Image processing; Metabolite; Acidic catecholamine metabolites; Elimination; Catecholamine; Thin layer chromatography; Carboxylic acid; Biogenic amine; Clinical biology; Image-processing analysis; Radical detection; Thin-layer chromatography; Quantitative analysis
• ISSN: 1615-9306; 1615-9314; 1615-9314
• DOI: 10.1002/jssc.201400550

A sensitive and convenient method for acidic catecholamine metabolites (including homovanillic acid, vanillylmandelic acid, 3,4‐dihydroxymandelic acid, and 3,4‐dihydroxyphenylacetic acid) determination was developed based on thin‐layer chromatography and image‐processing analysis. The metabolites were separated without a prederivatization step using reversed phase RP‐18W high‐performance plates. The mobile phase composition, detection, and quantification conditions were systematically investigated through several trials. The reaction with 2,2‐diphenyl‐1‐picrylhydrazyl radical allowed specific detection of acidic catecholamine metabolites with a high sensitivity and a wide linear range. The limit of detection and the limit of quantification were in the range of 13–103 and 18–120 ng/spot, respectively, in all cases. Mean recoveries determined were in the range 95–106% for all of the investigated compounds. The proposed method allowed rapid simultaneous determination of acidic catecholamine metabolites from spiked human urine sample.