Quantification of Dihydroxyacetone Phosphate (DHAP) in Human Red Blood Cells by HPLC-TripleTOF 5600™ Mass Spectrometer.

Document Type

Article

Publication Date

1-1-2016

Identifier

DOI: 10.1007/978-1-4939-3182-8_10

Abstract

Triosephosphate isomerase (TPI) is a glycolytic enzyme which catalyzes the interconversion between glyceraldehyde-3-phosphate (G3P) and dihydroxyacetone phosphate (DHAP). TPI deficiency results in accumulation of DHAP in human red blood cells and other tissues. The disease is characterized by congenital hemolytic anemia, and progressive neuromuscular dysfunction. The laboratory diagnosis is generally made by measurement of TPI activity in RBCs. Measurement of DHAP can be useful in further confirmation and follow-up of the disease. We developed HPLC/TOF-MS method for quantitation of DHAP in RBCs. The method involves simple protein precipitation, reverse phase C8 column chromatography, ion pairing with tributylamine, and long run time of 50 min to separate the two isomers (G3P and DHAP).

Journal Title

Methods in molecular biology (Clifton, N.J.)

Volume

1378

First Page

81

Last Page

86

MeSH Keywords

Blood Chemical Analysis; Chromatography, High Pressure Liquid; Dihydroxyacetone Phosphate; Erythrocytes; Humans; Mass Spectrometry

Keywords

Dihydroxyacetone phosphate; Production ion; Red blood cells; Tributylamine (ion pair reagent )

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