title: Colorimetric quantification of sucrose in presence of thermo-sensitive polymers present in aqueous two-phase systems.
abstract: The use of biodegradable material such as simple carbohydrates and recyclable material such as thermo-sensitive polymers is in need to develop a sustainable aqueous two-phase system (ATPS) for the purification of biomolecules. Accurate determination of sucrose concentration is important in liquid-liquid equilibrium (LLE) study of carbohydrate-based ATPS. The well-established phenol-sulfuric acid method has been widely employed in the measurement of carbohydrate concentration. However, the presence of thermo-sensitive polymers, which has a lower critical solution temperature (LCST) below room temperature, in carbohydrate samples could hamper the precision of spectrophotometric analysis due to the formation of two phases or cloudiness in the sample. Thus, the following modifications were made in an attempt to eliminate the interference occurred during conventional phenol-sulfuric acid assay.•The modified assay for sucrose quantification was performed at an ice-cold temperature throughout the reaction in order to avoid the interference from thermo-sensitive polymers.•This method required a sample volume of 3 μL and hence the volume of other reagents employed was also considerably reduced.•The absorbance was measured at 520 nm which allowed a longer linearity range (0.05-7.5%, w/v).
authors.affiliation: Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia.
authors.forename: Subbarayalu
authors.initials: S
authors.lastname: Ramalakshmi
authors.affiliation: Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia.
authors.forename: Chien Wei
authors.initials: CW
authors.lastname: Ooi
authors.affiliation: Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
authors.forename: Arbakariya B
authors.initials: AB
authors.lastname: Ariff
authors.affiliation: Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia ; Advanced Engineering Platform, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia.
authors.forename: Ramakrishnan Nagasundara
authors.initials: RN
authors.lastname: Ramanan
keywords: Aqueous two-phase system; Biomolecule purification; Colorimetric methods; Liquid–liquid equilibrium; Triton