Screening method for the simultaneous determination of allantoin and uric acid from dried blood spots

• Published in: Journal of pharmaceutical and biomedical analysis 2023-02, Vol.225, p.115222-115222, Article 115222
• Main Authors: Kopčil, Michal, Kanďár, Roman
• Format: Article
• Language: English
• Subjects: Allantoin; Allantoin - analysis; Biomarkers; Chromatography, High Pressure Liquid - methods; Dried blood spot; Dried Blood Spot Testing; Humans; Liquid chromatography; Oxidative Stress; Tandem Mass Spectrometry; Uric Acid
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2022.115222

Uric acid and its oxidation product allantoin are excellent biomarkers of oxidative stress in humans. Currently, there are high requirements not only for tests monitoring oxidative stress but also for screening laboratory tests in general. The highest demand is imposed on the simplest sampling, easy transport of the sample, and the shortest possible analysis time. The possible solution how to fulfil the requirements is sampling by dried blood spot technique with subsequent HPLC-MS/MS analysis. A fast, sensitive, and reliable HPLC-MS/MS method for the simultaneous determination of uric acid and allantoin from dried blood spots using stable isotopically labelled analogs as internal standards was developed. The separation took place in the reversed phase within 3 min, with protein precipitation and extraction in a one-step procedure. The analytical parameters of the method were satisfactory with an excellent linear range. The presented method was used to determine allantoin and uric acid levels in dried blood spot samples from 100 healthy volunteer donors. The median uric acid concentration in the cohort was 239.3 µmol/L and the median allantoin concentration was 5.6 µmol/L. The presented analytical protocol and method are suitable for screening and monitoring allantoin and uric acid levels as biomarkers of oxidative stress in clinical practice. [Display omitted] •Simple and non-invasive sample collection by dried blood spot technique.•Screening method for allantoin and uric acid determination was developed.•The analytical protocol is fast, simple, sensitive, and selective.•The median allantoin concentration was 5.6 µM.•The median uric acid concentration was 239.3 µM.