Analysis of Glycosyls Profile of Aroma Glycosides in Meili Grape by High Performance Liquid Chromatography-RefractiveIndex Detector with Amino Column / 分析化学

ABSTRACT

A high performance liquid chromatography method based on amino column to uncover the glycosyls of aroma glycosides in Meili grape was established.Meili grapes were sampled from Yangling, Shaanxi province, China.Aroma glycosides in grape skins were extracted by extraction buffer (0.1 mol/L Na2HPO4/NaH2PO4, pH 7.0, 13% ethanol (V/V)), then enzymatically hydrolysed by AR2000 enzyme preparation in citrate-phosphate buffer (pH 5.0), and the glycosyls liberated were isolated by amino column, and determined by high performance liquid chromatography-refractive index detector (HPLC-RID).The HPLC-RID experimental conditions were developed as follows amino column temperature 35℃, RID temperature 35℃, injection volume 20 μL, mobile phase acetonitrile/ethyl acetate/water=60∶25∶15 (V/V), flow rate 1 mL/min.The results showed that the reasonable linearity was achieved for rhamnose, xylose, arabinose, apiose and glucose (R2>0.996), with the detection limit (LOD) of 93-123 mg/L and quantitation limit (LOQ) of 309-409 mg/L.The relative standard deviation (RSD, n=10) of the peaks of each monosaccharide (5 g/L) was 2.3%-6.4%, and the recoveries ranged from 73.8% to 125.7%.The molar percentages of the aroma glycosides in Meili grape were 4.1%-6.1% for 6-O-α-L-rhamnopyranosyl-β-D-glucopyranoside, 2.3%-8.8% for 6-O-β-D-xylopyranosyl-β-D-glucopyranoside, 0.1%-3.9% for 6-O-α-L-arabinofuranosyl-β-D-glucopyranoside, 5.5%-9.8% for 6-O-α-L-apiofuranosyl-β-D-glucopyranoside, and 76.3%-86.8% for 6-O-β-D-glucopyranosyl-β-D-glucopyranoside.The contents of each aroma glycoside and the total glycosides in Meili grape didn′t show obvious relationship with berry maturity index during grape maturity.