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Simultaneous photometric microplate assay for free and total thiamine using gold nanoparticles and alkaline phosphatase

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Abstract

A photometric method is described for the determination of free and total thiamine (TH; vitamin B1) in food samples. It is based on the use of gold nanoparticles (AuNPs) which aggregate in the presence of TH owing to the interaction of its sulfur atom with the AuNPs. This is accompanied by a color change from wine red to purple-blue and an increase in absorbance at 590 nm. The effect is not observed for TH phosphates which, however, can be determined, as demonstrated for the example of TH pyrophosphate as a model ester, by treating them with alkaline phosphatase (ALP) which hydrolyses such esters. The use of two sample aliquots, one with and one without ALP, allows the determination of free and total TH, respectively. The dynamic range extends from 0.15 to 3.5 μM, and the detection limit is 54 nM of TH. The method has been applied to the analysis of spiked food samples and gave recoveries that ranged between 88.8 and 100.7 %.

Free thiamine can be directly determined by photometry at 590 nm as a result of the aggregation of gold nanoparticles (1st aliquot), whereas total thiamine requires the previous hydrolysis of the phosphate esters using alkaline phosphatase (2nd aliquot). Photograph: Gold nanoparticles in the absence and in the presence of thiamine (in duplicate).

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Acknowledgments

Authors gratefully acknowledge financial support from the MICINN (Grant No. CTQ2012-32941), from the Junta of Andalucía (Grant No.P09-FQM4933) and from the FEDER Program.

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Authors and Affiliations

  1. Analytical Chemistry Department, Institute of Fine Chemistry and Nanochemistry, University of Córdoba, Campus of Rabanales, Annex to Marie Curie Building, 14071, Córdoba, Spain

    María Ángeles Molina-Delgado, María Paz Aguilar-Caballos & Agustina Gómez-Hens

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  1. María Ángeles Molina-Delgado
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  2. María Paz Aguilar-Caballos
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  3. Agustina Gómez-Hens
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Correspondence to Agustina Gómez-Hens.

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Molina-Delgado, M.Á., Aguilar-Caballos, M.P. & Gómez-Hens, A. Simultaneous photometric microplate assay for free and total thiamine using gold nanoparticles and alkaline phosphatase. Microchim Acta 183, 1385–1390 (2016). https://doi.org/10.1007/s00604-016-1767-x

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