title: Improved evaporative light scattering detection for carbohydrate analysis.
abstract: Optimization and validation of evaporative light scattering detector (ELSD), aided by response surface methodology (RSM), has been developed for the liquid chromatography analysis of a wide molecular weight (MW) range of carbohydrates, including polysaccharides and oligosaccharides. Optimal experimental parameters for the ELSD detection were: 88.8°C evaporator temperature, 77.9°C nebulizer temperature and 1.1 standard litres per minute nitrogen flow rate. Optimal ELSD detection, used together with high performance size exclusion chromatography (HPSEC) of carbohydrates, gave a linear range from 250 to 1000 mg L(-1) (R(2)>0.998), with limits of detection and quantitation of 4.83-11.67 and 16.11-38.91 mg L(-1), respectively. Relative standard deviation was lower than 1.8% for intra-day and inter-day repeatability for apple pectin, inulin, verbascose, stachyose and raffinose. Recovery ranged from 103.7% to 118.3% for fructo-oligosaccharides, α-galacto-oligosaccharides and disaccharides. Optimized and validated ELSD detection is proposed for the analysis of high- to low-MW carbohydrates with high sensitivity, precision and accuracy.
authors.affiliation: Metabolism and Nutrition Department, Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN), Consejo Superior de Investigaciones Científicas (CSIC), José Antonio Novais 10, Ciudad Universitaria, ES-28040 Madrid, Spain.
authors.forename: Luis
authors.initials: L
authors.lastname: Condezo-Hoyos
authors.affiliation: Metabolism and Nutrition Department, Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN), Consejo Superior de Investigaciones Científicas (CSIC), José Antonio Novais 10, Ciudad Universitaria, ES-28040 Madrid, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Pérez-López
authors.affiliation: Metabolism and Nutrition Department, Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN), Consejo Superior de Investigaciones Científicas (CSIC), José Antonio Novais 10, Ciudad Universitaria, ES-28040 Madrid, Spain. Electronic address: pruperez@ictan.csic.es.
authors.forename: Pilar
authors.initials: P
authors.lastname: Rupérez
mesh_terms: D002241:Carbohydrates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008027:Light; D011134:Polysaccharides; D012542:Scattering, Radiation
chemical_list: D002241:Carbohydrates; D011134:Polysaccharides
keywords: Carbohydrate; ELSD; HPSEC; Oligosaccharide; Polysaccharide; RSM