食品科学 ›› 2023, Vol. 44 ›› Issue (22): 16-23.doi: 10.7506/spkx1002-6630-20221111-125

• 食品化学 • 上一篇    下一篇

中压制备色谱法分离黑果枸杞中2 个矮牵牛素花色苷及其对胰脂肪酶的抑制活性

米佳, 张渌淘, 禄璐, 魏佳仪, 金波, 罗青, 闫亚美, 曹有龙   

  1. (宁夏农林科学院枸杞科学研究所/国家枸杞工程技术研究中心,宁夏 银川 750002)
  • 出版日期:2023-11-25 发布日期:2023-12-13
  • 基金资助:
    国家自然科学基金地区科学基金项目(31860083);宁夏自然科学基金项目(2019AAC03288); 宁夏青年科技人才托举工程项目(NXKJTJGC2020048)

Separation and Pancreatic Lipase Inhibitory Activity of Two Petunidin Anthocyanins from Lycium ruthenicum Murr. by Preparative Medium-Pressure Liquid Chromatography

MI Jia, ZHANG Lutao, LU Lu, WEI Jiayi, JIN Bo, LUO Qing, YAN Yamei, CAO Youlong   

  1. (Institute of Wolfberry Engineering Technology, Ningxia Academy of Agriculture and Forestry Sciences/National Wolfberry Engineer Research Center, Yinchuan 750002, China)
  • Online:2023-11-25 Published:2023-12-13

摘要: 为了快速制备较高纯度的黑果枸杞花色苷,黑果枸杞花色苷粗提物首先用大孔树脂-中压制备色谱进行纯化,再用凝胶树脂-中压制备色谱分离获得2 个花色苷单体,采用超高效液相色谱-串联质谱(ultra-high performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)、核磁共振氢谱(1H nuclear magnetic resonance,1H NMR)对其结构进行鉴定,并对纯化物的胰脂肪酶抑制活性进行评价。结果表明:用D101大孔树脂-中压制备色谱对黑果枸杞花色苷进行纯化,所得纯化物中P1、P2的相对含量分别为18.22%和30.77%,继续采用Sephadex LH-20联合中压制备色谱分离纯化可得到相对含量分别为84.16%和81.48%的两个黑果枸杞花色苷组分P1和P2,经UPLC-MS/MS和1H NMR鉴定,2 个花色苷化合物分别为矮牵牛素-3-O-芸香糖(反式-对香豆酰基)-葡萄糖苷-5-O-葡萄糖苷和矮牵牛素-3-O-芸香糖(反式-对香豆酰基)-5-O-葡萄糖苷。P1和P2具有较好的胰脂肪酶抑制活性,其IC50值分别为0.250 mg/mL和0.224 mg/mL,且远低于花色苷粗提物和大孔树脂纯化物。研究为黑果枸杞花色苷利用提供一定基础。

关键词: 黑果枸杞;花色苷;矮牵牛素;中压制备色谱;大孔树脂;凝胶树脂

Abstract: Two anthocyanin monomers were separated from the crude anthocyanin extract from Lycium ruthenicum Murr. (LRM) by sequential preparative medium-pressure liquid chromatography (MPLC) on microporous and gel-type resins, and their structures were identified by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and nuclear magnetic resonance (1H NMR) spectroscopy. Their inhibitory activity against pancreatic lipase was also evaluated. Results indicated that the contents of anthocyanins P1 and P2 in the purified product obtained with macroporous resin D101 were 18.22% and 30.77% and were increased to 84.16% and 81.48% after chromatography on Sephadex LH-20, respectively. P1 and P2 were identified as petunidin 3-O-rutinoside(trans-p-coumarin)-glucoside-5-O-glucoside and petunidin 3-O-rutinoside(trans-p-coumarin)-5-O-glucoside by UPLC-MS/MS and 1H NMR, respectively. Both of them showed an excellent inhibitory activity against pancreatic lipase with half maximal inhibitory concentration (IC50) of 0.250 and 0.224 mg/mL, respectively, which were much lower than that of the crude anthocyanin extract and the macroporous resin D101 purified product. This work provides a basis for further research and utilization of anthocyanins from LRM.

Key words: Lycium ruthenicum Murr.; anthocyanin; petunidin; preparative medium-pressure liquid chromatography; macroporous resin; gel-type resin

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