Oligo Data

Description

Metabolites analysis on alpha-oligosaccharides in Tswana M5 mutants

Steps to reproduce

A rapid high-performance liquid chromatographic method with evaporative light scattering detection (HPLC-ELSD) analysis of alpha-galacto-oligosaccharides was conducted at the Central Analytical Facility (CAF) at Stellenbosch University, South Africa, using a modified procedure.35 The analysis was performed using a Waters Acquity UPLC BEH Amide column (1.7 μm, 2.1×100 mm) connected to Waters Acquity Ultra-Performance Liquid Chromatography (UPLC) coupled with an Evaporative Light Scattering Detector (UPLC ELSD) (Waters, Milford, MA, USA). The optimal ELS detector drift tube temperature was set at 50 °C, nitrogen gas was used as the nebulizing gas at 45 psi, and the gain was set to 200. The gradient program was set for two mobile phases. Phase A was set with 0.2% triethylamine and 0.2% ammonium hydroxide in water, while phase B was set with 0.2% triethylamine, 0.1 % 2-propanol, and 0.25% ammonium hydroxide in acetonitrile. For the optimization of separation conditions, standard solutions of the oligosaccharides: stachyose hydrate, verbascose analytical standard, Galactinol dihydrate, D-(+)-sucrose, and D-(+)-raffinose were prepared in 50:50 acetonitrile/water. A linear calibration curve (R2 = 0.998) was used to quantify the unknown samples. Solutions were filtered with KX Syringe filters (pore size 0.22 µm) and transferred to the vials. The injection volume was 5 µL, the flow rate was set at 0.3 mL /min with a column temperature of 50 °C and a total run time of 10 minutes. The analysis was performed in triplicate. The results were acquired, integrated and processed using Waters Empower Chromatography Data System.

Institutions

• Botswana University of Agriculture and Natural Resources

  Gaborone, Gaborone

Categories

Funders

• Organization for Women in Science for the Developing World

  Friuli Venezia Giulia, Trieste

Licence