基于氨基柱的高效液相色谱-示差折光检测器方法分析媚丽葡萄香气糖苷的糖基组成

建立了基于氨基柱的高效液相色谱分析媚丽葡萄香气糖苷的糖基组成的方法.媚丽葡萄果皮经提取缓冲液(0.1 mol/L Na2HPO4-NaH2PO4,pH 7.0,13%(V/V)乙醇)浸提,葡萄香气糖苷提取物在柠檬酸-磷酸盐缓冲液(pH 5.0)中用AR2000糖苷酶水解,释放出糖基,经氨基柱分离,用高效液相色谱-示差折光检测器(HPLC-RID)分析.色谱分析条件为:柱温35℃,RID温度35℃,进样量20 μL,流动相为乙腈-乙酸乙酯-水(60∶25∶15,V/V),流速1.0 mL/min.结果表明,鼠李糖、木糖、阿拉伯糖、芹菜糖和葡萄糖在线性范围内线性关系良好(R2>0.996),检出限为93~123 mg/L,定量限309~409 mg/L, 5 g/L各单糖的峰面积的精密度(n=10)为2.3%~6.4%;糖基样品的各单糖加标回收率在73.8%~125.7%之间.以糖基类别划分的媚丽葡萄各类香气糖苷的摩尔百分比为: 6-O-α-L-吡喃鼠李糖基-β-D-吡喃葡萄糖苷,4.1%~6.1%;6-O-β-D-吡喃木糖基-β-D-吡喃葡萄糖苷,2.3%~8.8%;6-O-α-L-呋喃阿拉伯糖基-β-D-吡喃葡萄糖苷,0.1%~3.9%;6-O-α-L-呋喃芹菜糖基-β-D-吡喃葡萄糖苷,5.5%~9.8%;6-O-β-D-吡喃葡萄糖基-β-D-吡喃葡萄糖苷,76.3%~86.8%.媚丽葡萄成熟过程中,各类香气糖苷的含量及其总量与浆果成熟度之间没有明显的相关性.

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

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.