title: Development and validation of an HPLC method to determine metabolites of 5-hydroxymethylfurfural (5-HMF).
abstract: The food component 5-hydroxymethylfurfural is supposed to have antioxidative properties and is therefore used as an acting agent in a novel anticancer infusion solution, named Karal®, and an oral supplementation. Previous studies showed that after oral and intravenous application, the substance is completely decomposed to its metabolites: 5-hydroxymethylfuroic acid, 2,5-furandicarboxylic acid, and N-(hydroxymethyl)furoyl glycine. The formation of a fourth metabolite, namely 5-sulphoxymethylfurfural, is still not clarified according to literature. Due to commercial unavailability, synthesis of 5-sulphoxymethylfurfural was conducted and a synthesis procedure for N-(hydroxymethyl)furoyl glycine had to be developed. Identification of the synthesised compounds was proven by LC-MS and NMR. An appropriate HPLC method was established to obtain good separation of the four possible metabolic substances and 5-hydroxymethylfurfural within 12 min via a HILIC column (150 × 4.6 mm, 5 μm) using a gradient grade system switching from mobile phase A (ACN/ammonium formate 100 mM, pH 2.35, 95:5, v/v) to mobile phase B (ACN/ammonium formate 100 mM, pH 2.35, 85:15, v/v). The procedure was afterward validated following ICH guidelines in terms of selectivity, linearity, precision, LOD, and LOQ.
authors.affiliation: Department of Pharmaceutical Chemistry, Institute of Pharmaceutical Sciences, Karl-Franzens University Graz, Graz, Austria.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Hardt-Stremayr
authors.forename: Marketa
authors.initials: M
authors.lastname: Bernaskova
authors.forename: Stefanie
authors.initials: S
authors.lastname: Hauser
authors.forename: Olaf
authors.initials: O
authors.lastname: Kunert
authors.forename: Xinghua
authors.initials: X
authors.lastname: Guo
authors.forename: Janette
authors.initials: J
authors.lastname: Stephan
authors.forename: Josef
authors.initials: J
authors.lastname: Spreitz
authors.forename: Ernst
authors.initials: E
authors.lastname: Lankmayr
authors.forename: Martin G
authors.initials: MG
authors.lastname: Schmid
authors.forename: Reinhold
authors.initials: R
authors.lastname: Wintersteiger
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005662:Furaldehyde / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: C008046:5-hydroxymethylfurfural; D005662:Furaldehyde