Trans-3,4-dideoxyglucone-3-ene (trans-3,4-DGE), a most reactive glucose degradation product in freshly heat sterilized glucose solutions

•Trans-3,4-dideoxyglucosone-3-ene (trans-3,4-DGE), a most reactive glucose degradation product, in heat sterilized dextrose 5% in water (D5W) was identified.•The trans-3,4-DGE was identified by reaction-directed fractionation of D5W, derivatization, and structure elucidation of the derivatives by MS and NMR.•The presence of trans-3,4-DGE in heat sterilized D5W was confirmed with authentic trans-3,4-DGE and NMR analysis of the reactive fraction. [Display omitted] In our study, one or more glucose degradation products (GDPs) in freshly heat sterilized dextrose 5% in water (D5W) were found to react with a drug candidate having a β-keto amide group (Compound A) to form several drug related compounds with the same molecular weight. However the previously identified GDPs did not react with Compound A to produce the observed adducts, indicating that unidentified GDP(s) reacted with Compound A to form these adducts. Our investigation by reaction-directed fractionation of the reactive D5W with HPLC led to the identification of the reactive GDP, trans-3,4-dideoxyglucosone-3-ene (trans-3,4-DGE), responsible for producing these reaction products. The trans-3,4-DGE was identified from its derivatives of dinitrophenylhydrazine (DNPH) and acetoacetanilide and confirmed by (1) admixing Compound A with authentic trans-3,4-DGE to produce the identical impurities as admixing with freshly heat sterilized D5W, and (2) NMR analysis of the reactive fraction of glucose solutions.