樟叶越橘的酚类成分:二级质谱裂解行为和神经保护作用  

Phenols from Vaccinium dunalianum Wight:Secondary Mass Spectrometry Fragmentation and Neuroprotective Effect

作　　者：周堂 王吉 乔温皓 林芷淇 朱红波 常月 胡炜彦 张荣平 陈兴龙 ZHOU Tang;WANG Ji;QIAO Wenhao;LIN Zhiqi;ZHU Hongbo;CHANG Yue;HU Weiyan;ZHANG Rongping;CHEN Xinglong(Yunnan Key Laboratory of Southern Medicine Utilization,College of Traditional Chinese Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China;Yunnan Key Laboratory of Pharmacology for Natural Products,School of Pharmaceutical Sciences,Kunming Medical University,Kunming 650500,China)

机构地区：[1]云南中医药大学中药学院,云南省南药可持续利用研究重点实验室,云南昆明650500 [2]昆明医科大学药学院,云南省天然药物药理重点实验室,云南昆明650500

出　　处：《食品科学》2024年第23期140-149,共10页Food Science

基　　金：云南省基础研究计划中医联合专项青年项目(202101AZ070001-113);“云岭学者”支持计划专项(YNWR-YLXZ-2019-019);云南省重点领域科技计划项目(202303AC100025);云南省南药可持续利用重点实验室开放课题(202105AG070012XS23089)。

摘　　要：以樟叶越橘嫩芽为研究对象,利用硅胶、小孔树脂、Sephadex LH-20和RP-18等分离材料,结合制备型和半制备型高效液相色谱,通过高分辨质谱和核磁等结构鉴定手段,从樟叶越橘的嫩芽水提取物中分离鉴定16个酚类化合物,包括7个熊果苷类、4个黄酮类、2个木脂素类和3个其他酚类化合物,其中化合物2~5、7和11~16为首次从越橘属植物中分离得到。二级质谱分析发现在负离子模式下糖苷类成分断裂糖苷键是其主要裂解途径,产生苷元部分或糖基部分的碎片,苷元部分或糖基碎片会继续丢失H_(2)O、CO、CHO或CH_(2)O等中性碎片,产生一系列子离子;黄酮或木脂素类成分丢失羟基、羟甲基、甲氧基等取代基,产生中性碎片或加合离子碎片;逆狄尔斯-阿尔德裂解在黄酮母核的裂解过程中也较为常见。利用H_(2)O_(2)损伤的SH-SY5Y细胞,评价了所有化合物的神经保护作用。结果表明化合物1~4、6、10和16对H_(2)O_(2)损伤SH-SY5Y细胞具有保护作用,其中化合物3(淫羊藿次苷F2)在5、10、20μmol/L剂量条件下其神经保护作用随浓度增加而提高,且存活率在3个剂量浓度条件下均超过80%,神经保护作用强于阳性药物表没食子儿茶素没食子酸酯。以上研究结果说明樟叶越橘含有丰富的酚类成分,这些成分表现出较强的神经保护作用,拓展了樟叶越橘嫩芽作为绿色健康食品的保健功能。In order to elucidate the health efficacy of Vaccinium dunalianum buds as a tea substitute and to reveal the material basis for its functionality,we studied the chemical constituents of V.dunalianum buds by using separation materials such as silica gel,MICHP-20p gel,Sephadex LH-20,and RP-18 combined with preparative and semi-preparative high performance liquid chromatography(HPLC),high-resolution mass spectrometry and nuclear magnetic resonance(NMR)spectroscopy.Totally 16 phenolic compounds were isolated and identified from the water extract of V.dunalianum buds,including seven arbutins,four flavonoids,two lignansand,and three other phenolic compounds,among which compounds 2–5,7,and 11–16 were isolated from the Vaccinium genus for the first time.Secondary mass spectrometry(MS/MS)analysis revealed that the major fragmentation pathway of glycosides in the negative ion mode was the breakage of glycosidic bonds to produce fragments of aglycone or glycosyl units.Then,the aglycone or glycosyl units continued to lose neutral fragments such as H_(2)O,CO,CHO,and CH_(2)O to yield a series of sub-ions.Flavonoids or lignans lost their substituents such as hydroxyl,hydroxymethyl,and methoxyl groups to produce neutral fragments or additional ions.Besides,Retro-Diels-Alder(RDA)fragmentation was also common in the cleavage of the flavonoid skeleton.Compounds 1–4,6,10 and 16 had protective effects on SH-SY5Y cells damaged by H_(2)O_(2).The neuroprotective effect of compound 3(Icariside F2)increased with increasing its concentration at 5,10,and 20μmol/L,and the survival rate exceeded 80%at all three concentrations.The neuroprotective effect was stronger than that of the positive drug epigallocatechin gallate.The above findings indicated that V.dunalianum buds was rich in phenolic compounds with obvious neuroprotective effects,which would expand the health functions of V.dunalianum buds as green health foods.

关 键 词：樟叶越橘 酚类成分 二级质谱裂解行为 神经保护作用 

分 类 号：R28[医药卫生—中药学]