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 1000mgL−1 (R2>0.998), with limits of detection and quantitation of 4.83–11.67 and 16.11–38.91mgL−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.
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