title: Quantification of Dihydroxyacetone Phosphate (DHAP) in Human Red Blood Cells by HPLC-TripleTOF 5600™ Mass Spectrometer.
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).
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Mercy Hospitals and Clinics, 2401 Gillham Road, Kansas City, MO, 64108, USA.
authors.forename: Shuang
authors.initials: S
authors.lastname: Deng
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Mercy Hospitals and Clinics, 2401 Gillham Road, Kansas City, MO, 64108, USA.
authors.forename: David
authors.initials: D
authors.lastname: Scott
authors.affiliation: Department of Pediatrics, Children's Mercy Hospitals and Clinics, Kansas City, MO, USA.
authors.forename: Douglas
authors.initials: D
authors.lastname: Myers
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Mercy Hospitals and Clinics, 2401 Gillham Road, Kansas City, MO, 64108, USA. ugarg@cmh.edu.|University of Missouri School of Medicine, Kansas City, MO, USA. ugarg@cmh.edu.
authors.forename: Uttam
authors.initials: U
authors.lastname: Garg
mesh_terms: D001774:Blood Chemical Analysis / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004099:Dihydroxyacetone Phosphate / Q000097:blood*; D004912:Erythrocytes / Q000737:chemistry*; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*
chemical_list: D004099:Dihydroxyacetone Phosphate
keywords: Dihydroxyacetone phosphate; Production ion; Red blood cells; Tributylamine (ion pair reagent )