Determination of selected alpha-keto acids in dried blood samples using HPLC with fluorescence detection
Článekpeer-reviewedpublishedDatum publikování
2022
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Elsevier Science Inc.
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The determination of alpha-keto acids derived from amino acids is currently the most reliable approach for the diagnosis of some congenital metabolic diseases. An HPLC method for the simultaneous measurement of selected alpha-keto acids in dried blood samples has been developed and evaluated. Blood spot samples from a group of healthy blood donors were collected onto #903 Specimen Collection Paper. Prior the separation, the alpha-keto acids were derivatized with 1,2-diamino-4,5-dimethoxybenzene to the corresponding 3-substituted-6,7-dimethoxy-2 (1 H)-quinoxalinol derivatives. For the separation, a reverse-phase column LichroCart 125-4, Purospher RP-18e, 5 mu m, was used. The mixture of 25% ACN in deionized water (mobile phase A) and 100% ACN (mobile phase B) were used for a gradient elution of alpha-keto acids derivatives. Analytical performance of this method is satisfactory for all alpha-keto acids. The intra-assay and inter-assay coefficients were below 10% and recoveries were close to 100%. We have developed relatively simple, rapid, selective and sufficiently sensitive HPLC method with fluorescence detection for the determination of selected alpha-keto acids in dried blood samples. The presented method is suitable for clinical testing purposes.
Rozsah stran
p. 114738
ISSN
0731-7085
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Journal of Pharmaceutical and Biomedical Analysis, volume 214, issue: May
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https://www.sciencedirect.com/science/article/pii/S0731708522001595?via%3Dihub
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alpha-Keto acids, dried blood spot, HPLC with fluorescence detection, 1,2-Diamino-4,5-dimethoxybenzene, α-oxokyseliny, suchá kapka krve, HPLC s fluorescenční detekcí, 1,2-diamino-4,5-dimethoxybenzen