为提高核桃种皮多酚粗提物的纯度并明晰核桃种皮多酚的种类,采用大孔吸附树脂对核桃种皮多酚进行富集,筛选出最佳树脂,优化分离工艺条件,并对其富集产物进行鉴定分析。结果表明:AB-8树脂最适用于核桃种皮多酚的富集。最佳工艺条件为:核桃种皮多酚溶液上样质量浓度2 mg/mL, pH 3.0, 静态吸附时间3 h,解吸液为体积分数70%的乙醇溶液,动态吸附最大上样量为14 BV,洗脱液用量为4 BV。在此条件下,核桃种皮多酚的纯度可由24.24%提高到70.52%。用超高效液相色谱串联四级杆飞行时间质谱对纯化后的核桃种皮多酚类物质进行鉴定,确认鞣花酸、鞣花酰基葡萄糖异构体、二鞣花酰基葡萄糖、三没食子酰基鞣花酰基葡萄糖、四没食子酰基葡萄糖、木麻黄素/木麻黄鞣亭异构体、Glansrin C异构体、鞣花酸己糖异构体、鞣花酸戊糖异构体及Glansreginin A等30种多酚类组分。该研究表明大孔吸附树脂可作为核桃多酚初步分离的介质,其对鞣花酸及其衍生物等功效成分有较好的保留。
The macroporous adsorption resins were used to enrich the walnut pellicle polyphenols and the separation process conditions were optimized, the phenolic compounds were then identified. The results showed that the AB-8 resin was the ideal resin for enriching crude walnut pellicle polyphenols. The ideal enrichment parameters were as follows: sample concentration was 2.00 mg/mL at pH 3.0 with the static adsorption time for 3 h, meanwhile 70% ethanol was used as the eluent solvent. Dynamic adsorption study showed that the maximum loading amount was 14 BV and the amount of eluent was 4 BV. Under above conditions, the purity of walnut pellicle polyphenols was increased from 24.24% to 70.52%. The walnut pellicle polyphenols were identified using the Ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-TOF-MS/MS). There were more than 30 kinds of polyphenols compounds were tentatively identified including ellagic acid, HHDP-glucose isomer, Di-HHDP-glucose, trigalloyl-HHDP glucose, teragalloyl-glucose, casuarinin/casuarinin isomer, glansrin C isomer, ellagic acid hexose isomer, ellagic acid pentose isomer, glansreginin A. This research indicates that macroporous adsorption resin could act as a preliminary separation media for refined walnut polyphenols containing mainly ellagic acid and its derivatives.
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