A Simple Analytical Method for High-Throughput Screening of Major Sugars from Soybean by Normal-Phase HPLC with Evaporative Light Scattering Detection

Author:

Valliyodan Babu1,Shi Haiying1,Nguyen Henry T.1

Affiliation:

1. National Center for Soybean Biotechnology and Division of Plant Sciences, University of Missouri, Columbia, MO 65211, USA

Abstract

This paper presents a simple analytical method for determining sugars in soybean (Glycine max (L.) Merr.) tissues. Sample preparation was modified from several early published methods. High-performance liquid chromatography (HPLC) equipped with an evaporative light scattering detector (ELSD) was used to separate, identify, and quantify seven sugars, including glucose, galactose, fructose, sucrose, melibiose, raffinose, and stachyose. Two mobile phases were programed into a gradient elution. Mobile phase A is pure water and mobile phase B is a mixture of acetonitrile and acetone 75 : 25 (v/v). Total chromatographic retention time is less than 20 minutes. This method has been validated for detection limit, calibration range, and intraday and interday repeatability. This method has been used analyzing more than 5000 soybean samples in the experiments studying natural genetic variation of sugar contents and components in soybean seeds and other tissues.

Publisher

Hindawi Limited

Subject

General Earth and Planetary Sciences,General Engineering,General Environmental Science

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