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.

Determination of metabolites of 5‐hydroxymethylfurfural in human urine after oral application

“…In addition 5‐HMF also shows antioxidative properties, is able to bind free radicals and it is supposed to protect from hypoxia . It is part as an acting agent in an anti‐cancer infusion solution, termed Karal®, being in Phase II clinical trials at the current stage, and shows capacity to be used in targeted cancer therapy . Further 5‐HMF can be administered to patients orally as Sanopal®.…”

“…Analysis of the metabolic substances has been conducted by many different ways before, including LC‐UV methods , but also GC‐MS and LC‐MS were used. In our previous work, a rapid and cost‐effective method was already established . This HPLC method was adapted for determination of the compounds also in urine samples after purification prior to use.…”

Determination of furanic aldehydes in sugarcane bagasse by high‐performance liquid chromatography with pulsed amperometric detection using a modified electrode with nickel nanoparticles

“…A wide array of works found in the literature devoted to the determination of furanic aldehydes in different matrices has usually applied chromatographic techniques . Among these techniques HPLC with UV , UV ‐ diode array , UV visible , diode array , refractive index , evaporative light scattering detectors , MS and MS/MS detection are the most widely used.…”

An improved method for the determination of 5‐hydroxymethylfurfural in Shenfu injection by direct analysis in real time‐quadrupole time‐of‐flight mass spectrometry

“…It is essential that the appropriate methods should be established to ensure the quality of injected medicine, and a pharmacopeia limit test for 5‐HMF and related substances was often performed by ultraviolet spectrophotometry (UV). Recently, analysis of 5‐HMF in various substrates such as honey, urine or human plasma by several methods, including high performance liquid chromatography (HPLC) with UV detector /mass spectrometry, gas chromatography–mass spectrometry (GC‐MS) and micellar electrokinetic capillary chromatography (MEKC) technique, have been reported. However, quantification methods of 5‐HMF in the commercial injections were less reported.…”