Highly-substituted benzhydrylamine resin: An alternative chromatographic support for ganglioside and neutral glycosphingolipid purification

Summary Benzhydrylamine resin (BHAR) is a copolymer of (styrene-1% divinylbenzene) containing phenylmethylamine groups and commonly used as a solid support for peptide synthesis in organic solvents. We have demonstrated that a larger number of NH₃ ⁺ groups distributed throughout the BHAR matrix improves bead solvation under more polar conditions. As this characteristic might allow this aminated-polymer to be used as a stationary phase for liquid chromatography, a hyghly substituted BHAR (2.4 mmol.g⁻¹ of amine groups) was synthesized under forceful conditions. Neutral glycosphingolipids and negatively charged gangliosides and sulfatide obtained from rat brain extracts were successfully purified and fractionated in this BHAR batch by single-step, reverse-phase anion-exchange chromatography. Additionally, the anion-exchange potential of BHAR was also compared to that of commercial dimethylaminoethyl (DEAE)-Sephadex A25 resin.