Fast and accurate diagnosis of G6PD deficiency via NADPH monitoring using sequential injection analysis coupled with electrochemical detection
Issued Date
2026-01-01
Resource Type
ISSN
00399140
Scopus ID
2-s2.0-105014617980
Pubmed ID
40902521
Journal Title
Talanta
Volume
297
Rights Holder(s)
SCOPUS
Bibliographic Citation
Talanta Vol.297 (2026)
Suggested Citation
Jandahong P., Charoenkitamorn K., Hongwitayakorn A., Chaneam S., Boonyuen U. Fast and accurate diagnosis of G6PD deficiency via NADPH monitoring using sequential injection analysis coupled with electrochemical detection. Talanta Vol.297 (2026). doi:10.1016/j.talanta.2025.128738 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/114619
Title
Fast and accurate diagnosis of G6PD deficiency via NADPH monitoring using sequential injection analysis coupled with electrochemical detection
Corresponding Author(s)
Other Contributor(s)
Abstract
A rapid and automated determination of nicotinamide adenine dinucleotide phosphate (NADPH) is proposed and applied to the evaluation of glucose-6-phosphate dehydrogenase (G6PD) deficiency in real samples. To this end, a sequential injection analyzer with electrochemical detection (SIA-ECD) is proposed with 0.1 mol L<sup>−1</sup> Tris-HCl (pH 8.0) as a supporting electrolyte and carrier solution. For sensitivity enhancement, cobalt(II) phthalocyanine (CoPc) was used to modify a screen-printed graphene electrode (SPGE), acting as a transducer. Scanning electron microscopy, cyclic voltammetry, and electrochemical impedance spectroscopy were employed to characterize the morphology and electrochemical properties of the modified CoPc/SPGE. Chronoamperometry was used to determine NADPH levels by measuring the increase in oxidation current. The SIA-ECD system was optimized with respect to the CoPc concentration, applied potential, sample volume, reagent volume, and flow rate towards the detector. The innovation provided a wide linear range from 10.0 μM to 1.50 mM and a limit of detection at 4.14 μmolL<sup>−1</sup>. It demonstrated good reproducibility with %RSD of 4.94 from 10 different electrodes. In addition, the fabricated electrode provided consistent signals across 12 injections (RSD = 1.2 %). For real sample analysis, the activity of non-mutant enzymes was compared to that of 6 mutant samples, revealing enzyme activity ranging from 12.5 ± 0.5 to 50.8 ± 3.0 %, which is in good agreement to the reference spectrometric method. No matrix interference was observed, as demonstrated by recovery ranging from 81.1 % to 107.7 %. The proposed procedure may process 120 samples per hour. To the best of the authors knowledge, this is the first procedure combining flow analysis with G6PD evaluation by monitoring NADPH levels.