摘要
【目的】采用溶剂萃取法提取紫皮豇豆中的总多酚,多酚提取物经大孔树脂分离纯化,再利用高效液相色谱(HPLC)分析其多酚类化合物的组成及含量,为紫皮豇豆中多酚类物质的提取、分析提供理论依据。【方法】以总多酚提取量为考察指标,采用单因素和正交试验,分析提取溶剂、料液比、温度和pH对紫皮豇豆总多酚提取量的影响,再通过Sephadex LH-20大孔树脂对粗提物进行分离纯化,并建立紫皮豇豆中6种多酚类组分的HPLC分析方法,采用外标法定量。【结果】紫皮豇豆中总多酚的最佳提取工艺为:经料液比1∶5(m∶V)的80%丙酮水溶液,在4℃、pH 6条件下,提取3次,每次提取5 h时,所得的总酚含量最高为614 mg(GAE)·100 g^-1。在优化的试验条件下,没食子酸、没食子儿茶素、绿原酸、芦丁、p-香豆酸和阿魏酸等6种多酚单体在相应的浓度范围内具有良好的线性关系,相关系数(r)为0.9959~0.9999,方法检出限和定量限分别为0.1~1 mg·100 g^-1和0.2~2.5 mg·100 g^-1;加标试验平均回收率74.3~107%,相对标准偏差0.4~8.6%。经HPLC分析,紫皮豇豆中6种多酚单体均有检出,含量在0.46~6.5 mg·100 g^-1,其中含量最高的是芦丁。【结论】采用80%丙酮水溶液提取紫皮豇豆总多酚,并利用HPLC法测定紫皮豇豆中几种单体酚的方法提取率高、准确性好,适用于紫皮豇豆中多酚类化合物的提取及定性定量分析。
【Objective】Polyphenol compounds in purple cowpeas were solvent-extracted,purified,and then chemically analyzed by HPLC.【Method】Single factor and orthogonal experiments were conducted to optimize the polyphenol extraction from purple cowpeas.Using polyphenol yield as the criterion,effects of solvent,solid-liquid ratio,temperature,and pH applied for the process were evaluated for optimization.The crude extract was purified using a Sephadex LH-20 macroporous resin.HPLC with external standards was employed for the chemical determination on the purified product.【Result】The optimized process applied a peas-to-solvent ratio of 1∶5(m∶V)with an 80% acetone solution to extract 3 times at 4℃ and pH 6 for 5 h per cycle to reach a maximum total polyphenols content of the gallic acid equivalent of 614 mg·100 g^-1.The correlation coefficients on the linear functions of the 6 polyphenols against their respective standards ranged from 0.9959 to 0.9999 with a limit of detection(LOD)of 0.1~1.0 mg·100 g^-1 and that of quantitation 0.2~2.5 mg·100 g^-1 on the HPLC test.The average recoveries on these compounds ranged from 74.3% to 107% with a relative standard deviation(RSDs)of 0.4~8.6%.The 6 polyphenols in the purple cowpeas thus determined averaged between 0.46 mg·100 g^-1 and 6.5 mg·100 g^-1,with rutin being the most abundant.ing the most abundant.【Conclusion】The polyphenol extraction from purple cowpeas was optimized to deliver high yields on 6 monomer phenols,which could be accurately determined quantitatively by HPLC.
作者
吴琼
王明月
葛会林
马晨
吕岱竹
WU Qiong;WANG Mingyue;GE Huilin;MA Chen;LYU Daizhu(Analysis&Testing Center,Chinese Academy of Tropical Agricultural Sciences/Agricultural Ministry Laboratory for Tropical Crop Product Quality and Safety Risk Assessment/Hainan Provincial Key Laboratory of Quality and Safety for Tropical Fruits and Vegetables,Haikou,Hainan 571101,China)
出处
《福建农业学报》
CAS
CSCD
北大核心
2020年第6期673-681,共9页
Fujian Journal of Agricultural Sciences
基金
海南省自然科学基金(No.318MS089)
国家农产品质量安全风险评估项目(GJFP201900605)。
关键词
紫皮豇豆
多酚
提取纯化
成分分析
Purple cowpea
total polyphenols
extraction and purification
composition analysis
