Analysis of Sugars by HPLC-ELSD

Download PDF August 25, 2017

Introduction

Sugar Analysis HPLC
PU- 2089

An evaporative light scattering detector (ELSD) is a universal HPLC detector whose detection principle is based on the phenomenon of light scattering, which occurs from particles of residual non-volatile components after removing the volatile mobile phase using a combination of heat and gas nebulization.  The ELSD detector uses an LED as the light source and a photomultiplier to measure change in intensity as the analyte passes through the optical path. Compounds that are typically measured using a refractive index detector or a short wavelength of a UV detector may be measured with increased sensitivity or a more stable baseline using ELSD.

In this application note, monosaccharides and disaccharides were analyzed using ELSD from a separation using a polymer NH2 column in HILIC mode.

Experimental

Instrument

PumpPU-2089S
AutosamplerAS-2057
Column OvenCO-2060
DetectorELSD-2040

Method

ColumnAsahiPak NH2P-50 4E
(4.6 ID x 250 L mm)
EluentH2O/ACN 25:75
Flow Rate1.0 mL/min
Column Temperature30 deg C
ELSD ParametersNebulizer temp. 30 deg C
Evaporator temp. 30 deg C
Gas flow rate 1.4 SLM
Injection Volume10uL
Standard SamplesRhamnose, Fructose, GLucose, Sucrose, Maltose
1.0mg/mL in H2O/ACN 50:50

Keywords

Sugar, HILIC, Polymer NH2 column, ELSD, Rhamnose, Fructose, Glucose, Sucrose, Maltose, monosaccharides

Results

Figure 1. Chromatogram of a standard mixture of monosaccharides and disaccharides.

Figure 1. Chromatogram of a standard mixture of sugars.
1: Rhamnose, 2: Fructose, 3: Glucose, 4: Sucrose, 5: Maltose

About the Author

Toshifumi Uchiyama is a member of the electronic spectroscopy team located at the JASCO main laboratory in Tokyo