富硒葛粉中硒形态分析及其对脂多糖诱导的AML12细胞氧化损伤的影响  

Determination of selenium species in Se-enriched puerarin powder and its effect on oxidative damage of AML12 cells induced by Lipopolysaccharide

作　　者：陆秋艳[1] 张小鸿 张文婷[1] 李丽林[1] LU Qiuyan;ZHANG Xiaohong;ZHANG Wenting;LI Lilin(Fujian Center for Disease Control and Prevention,Fujian Provincial Key Laboratory of Zoonosis,Fuzhou,350012,China)

机构地区：[1]福建省疾病预防控制中心,福建省人兽共患病研究重点实验室,福州350012

出　　处：《环境化学》2023年第10期3603-3607,共5页Environmental Chemistry

基　　金：福建省卫生健康科研人才培养项目医学创新课题(2019-CXB-14)资助.

摘　　要：建立了富硒葛粉中硒酸根[Se(Ⅵ)]、亚硒酸根[Se(Ⅳ)]、硒代胱氨酸(SeCys_(2))、甲基硒代半胱氨酸(MeSeCys)、硒代蛋氨酸(SeMet)、硒代乙硫氨酸(SeEt)等6种硒形态的检测方法并分析了主要硒形态的抗氧化损伤功能0.1 mol·L^(−1)的NaOH溶液在90℃下提取样品30 min,加入蛋白酶XIV、脂肪酶、淀粉酶,37℃水浴酶解18 h,高效液相色谱-电感耦合等离子体质谱联用(HPLC-ICP-MS)快速分离测定6种硒形态.分析主要硒形态对脂多糖(LPS)诱导的AML12细胞氧化损伤的影响.结果表明,6种硒形态的检出限为0.017—0.053 mg·kg^(−1),标准曲线的相关系数均在0.9998以上.该前处理方法样品的加标回收率在71.2%—117.3%,相对标准偏差均在10%以内.与模型组相比,1μmol·L^(−1)硒代蛋氨酸处理组脂多糖诱导的AML12细胞活力显著上升(P<0.01),丙二醛(MDA)含量下降30.1%(P<0.01),超氧化物歧化酶(SOD)活性增加43.9%(P<0.01),谷胱甘肽过氧化物酶(GSHx)活性增加44.7%(P<0.01).硒代蛋氨酸对LPS诱导的AML12细胞氧化损伤具有保护作用,可增强植物的抗氧化损伤能力.The objective of this study was to establish a method for the detection of six selenium species in Se-enriched puerarin powder,including Se(Ⅵ),Se(Ⅳ),SeCys_(2),MeSeCys,SeMet and SeEt.Also this study was to analyze the antioxidant damage of the main selenium species.The sample was extracted with 0.1 mol·L^(−1)NaOH solution at 90℃for 30 min,then protease XIV,lipase and amylase were added.And enzymolysis was performed at 37℃for 18 h in water bath.Six selenium species were rapidly detected by high performance liquid chromatography-inductively coupled plasma-mass spectrometry(HPLC-ICP-MS).The effect of main selenium species on the oxidative damage of AML12 cells induced by Lipopolysaccharide(LPS)was measured.The results showed that the detection limits for the six selenium species were 0.017—0.053 mg·kg^(−1),and the correlation coefficients of the standard curves were all above 0.9998.The spiked recoveries of the samples determined by this pretreatment method were all between 71.2%—117.3%,and the relative standard deviations were all within 10%.Compared to the model group,the activity of AML12 cells induced by LPS increased significantly(P<0.01)after being applied to SeMet at a concentration of 1μmol∙L^(−1),the content of Malondialdehyde(MDA)decreased by 30.1%(P<0.01),the activity of Superoxide Dismutase(SOD)increased by 43.9%(P<0.01),and the activity of Glutathione Peroxidase(GSHx)increased by 44.7%(P<0.01).The results suggest that SeMet has a protective effect on LPS induced oxidative damage in AML12 cells.So it can enhance the ability of plants on antioxidant damage.

关 键 词：硒 形态分析 高效液相色谱-电感耦合等离子体质谱联用 抗氧化损伤 

分 类 号：TS201.2[轻工技术与工程—食品科学] R151[轻工技术与工程—食品科学与工程]