Effects of Selenium Dioxide on Apoptosis, Bcl-2 and P53 Expression, Intracellular Reactive Oxygen Species and Calcium Level in Three Human Lung Cancer Cell Lines

To evaluate the anti-tumor effects of SeO2 and its mechanisms on three human lung cancer cell lines. Methods: Three lung cancer cells A549, GLC-82 and PG were treated with 3-30 μmol/L SeO2. Flow cytometry was used to detect apoptosis, and analyze the changes of expression of p53 and Bcl-2, as well as ROS and Ca^2+ level within cells. Results:SeO2 markedly inhibited cell proliferation and viability, and prompted apoptosis after 48 h treatment. SeO2 at 10 μmol/L induced 47.8% apoptosis in A549 cells, 40.8% in GLC-82 cells, 18.2% in PG cells. SeO2 at 30 μmol/L induced 37.8% apoposis in PG cells,but did not increase apoptotic raes in other two cells. SeO2 could down-regulate the mean fluorescent intensity of Bcl-2 from 65.8 to 9.6 in A549, but not in GLC-82 and in PG, cells, up-regulate wild type p53 level in all three cells. SeO2 decreased the ROS and Ca^2+ level markedly within three tested cells. Conclusion: SeO2 showed anti-tumor effect via apoptosis pathway in three lung cancer cell lines. The decrease of ROS and Ca^2+ level within cells as well as regulation of Bcl-2 and p53 expression may play important roles in above apoptotic procedure.

Species and distribution of inorganic selenium in the Bohai Sea

For studying geochemical behavior of selenium, a marine investigation was conducted in September 1998 cruise in the Bohai Sea. Horizontal and vertical distributions of inorganic selenium and selenite were studied. The inorganic selenium varied from 0.73 to 2.41 nmol/L, with an average of 1.31 nmol/L; while selenite varied from 0.18 to 0.72 nmol/L with an average of 0.47 nmol/L. The average ratio of selenite to selenate was 0.68, and selenate was the predominant species of inorganic selenium in the most study area. Concentrations of inorganic selenium and selenite decreased with distance from the coast. No apparent variations of the concentration between surface and bottom was shown. There were two wa- ter masses in the area, one from the Huanghe (Yellow) River and other from the Yellow Sea. Of the two, Huanghe River was the major source of inorganic selenium.

Effect of inorganic arsenic in paddy soil on the migration and transformation of selenium species in rice plants

Selenium(Se)in paddy rice is one of the significant sources of human Se nutrition.However,the effect of arsenic(As)pollution in soil on the translocation of Se species in rice plants is unclear.In this research,a pot experiment was designed to examine the effect of the addition of 50 mg As/kg soil as arsenite or arsenate on the migration of Se species from soil to indica Minghui 63 and Luyoumingzhan.The results showed that the antagonism between inorganic As and Se was closely related to the rice cultivar and Se oxidation state in soil.Relative to the standalone selenate treatment,arsenite significantly(p<0.05)decreased the accumulation of selenocystine,selenomethionine and selenate in the roots,stems,sheaths,leaves,brans and kernels of both cultivars by 21.4%-100.0%,40.0%-100.0%,41.0%-100%,5.4%-96.3%,11.3%-100.0% and 26.2%-39.7% respectively,except for selenocystine in the kernels of indica Minghui 63 and selenomethionine in the leaves of indica Minghui 63 and the stems of indica Luyoumingzhan.Arsenate also decreased(p<0.05)the accumulation of selenocystine,selenomethionine and selenate in the roots,stems,brans and kernels of both cultivars by 34.9%-100.0%,30.2%-100.0%,11.3%-100.0% and 5.6%-39.6%respectively,except for selenate in the stems of indica Minghui 63.However,relative to the standalone selenite treatment,arsenite and arsenate decreased(p<0.05)the accumulation of selenocystine,selenomethionine and selenite only in the roots of indica Minghui 63 by 45.5%-100.0%.Our results suggested that arsenite and arsenate had better antagonism toward Se species in selenate-added soil than that in selenite-added soil;moreover,arsenite had a higher inhibiting effect on the accumulation of Se species than arsenate.

超声波辅助酶提取结合高效液相色谱-电感耦合等离子体质谱法测定富硒蔬菜中硒形态

该研究建立了利用高效液相色谱-电感耦合等离子体质谱法(high performance liquid chromatography-inductively coupled plasma-mass spectrometry,HPLC-ICP-MS)同时测定富硒蔬菜中SeO_(4)^(2-)、SeO_(3)^(2-)、SeMet、SeCys 2、MeSeCys等5种硒形态化合物含量的方法。蔬菜样品0.2 g加入4 mg蛋白酶XIV,加入5.00 mL pH为7.5的水溶液在37℃超声波提取80 min,微孔滤膜过滤,利用Hamilton PRP-X100阴离子交换柱,以25 mmol/L柠檬酸溶液(pH 6.0)为流动相,可在7 min内完成5种硒形态的完全分离。SeO_(4)^(2-)、SeO_(3)^(2-)、SeMet、SeCys 2、MeSeCys的线性相关系数为0.9993~0.9995,回收率范围为96.3%~104.2%,相对标准偏差小于7.6%。实验结果表明该法操作简便、快速、精密度及准确度良好,适用于富硒蔬菜中硒的形态分析。

梯度洗脱LC-ICP/MS法测定5种硒形态

建立一种梯度洗脱液相色谱-电感耦合等离子体质谱法(Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometry,LC-ICP/MS)分析硒代胱氨酸、甲基硒代半胱氨酸、亚硒酸根、硒代蛋氨酸、硒酸根5种硒形态。样品通过Hamilton PRP-X100阴离子色谱柱,分别以流动相A 40 mmol·L^(-1)磷酸氢二铵+5 mmol·L^(-1)柠檬酸(pH=4.6)和流动相B 60 mmol·L^(-1)磷酸氢二铵+5 mmol·L^(-1)柠檬酸(pH=4.6)梯度洗脱5种硒形态,流速为1.0 mL·min^(-1),切换时间分别为3.0 min和6.4 min,ICP/MS检测,外标法定量。5种硒形态在8.5 min内检测完毕;0.5~200.0μg·L^(-1)相关系数r>0.999,最低检测质量浓度为0.10~0.39μg·L^(-1)。相较于《生活饮用水标准检验方法第6部分:金属和类金属指标》(GB/T 5750.6-2023)硒形态分析法,该方法将分析时间缩短了46.8%,5种形态峰分离度更好、灵敏度更高。

Preliminary Study of Selenium (Se) Toxicity in Human Prostate Carcinoma (PC3) Cells with the Overexpression of Selenocysteine Synthase (SecS) Gene

Selenium (Se) is a trace element required for normal body function. Its supplementation of human diet at standard optimum amount prevents oxidative damages in cells and could be a viable method in the prevention of diseases related to DNA damage, including cancer, neurodegenerative diseases and aging. While Se anticancer properties have been linked to its ability to remove excess Reactive Oxygen Species (ROS) in cells, the underlying molecular mechanism remains unknown. Recent studies have shown that the removal of ROS alone cannot account for Se anticancer properties. To really comprehend the molecular basis of Se anticancer properties, current researches now focus on the metabolism of Se in the cell, especially Se-containing amino acids. Selenocysteine (Sec) is a novel amino acid and one of the selenium-containing compounds in the cell. It is essential in the maintenance of the integrity of its parent proteins, some of which include enzymes such as Glutathione Peroxidases (GPXs) and Thioredoxin Reductases (TrXs). We propose in this study that the overproduction of Sec via the overexpression of Selenocysteine synthase (SecS) gene and Se supplementation induced cell death in Prostate Carcinoma (PC-3) cells. Although the mechanism underlying the cell death induction is unknown, we propose it could be due to the random incorporation of Sec into proteins at high concentration, causing premature protein degradation and cell death. The outcome of this study showed that increasing the concentration of intracellular Se-containing amino acids may provide important clinical implications for the treatment of cancer.

高效液相色谱-原子荧光光谱联用技术测定富硒鸡蛋中的硒形态

为监测市场上富硒鸡蛋中蛋清及蛋黄中硒形态的种类及分布规律,优化建立了一种高效液相色谱-氢化物发生-原子荧光光谱联用技术测定富硒鸡蛋蛋清及蛋黄粉中硒形态的方法。随机选取富硒鸡蛋样品,进行蛋清蛋黄分离、冷冻干燥及蛋黄脱脂处理,处理后样品经Tris-HCl缓冲液(5 mmol/L,pH=7.5) 55℃恒温振荡、酶解提取,高速离心机(10 000 r/min)离心10 min,上清液过0.22μm有机滤膜后采用高效液相色谱-氢化物发生-原子荧光光谱联用技术进行测定,外标法定量。结果表明,能在11 min内实现5种硒形态的基线分离,且各种硒形态的线性相关系数(r)均大于0.999 4,其检出限在0.5~3.0μg/L,回收率在81.6%~110%,可以满足检测需求,按照实验方法测定市场上富硒鸡蛋中的硒形态,方法灵敏度高、准确性好,且测得的富硒鸡蛋蛋清样品中硒形态主要成分为硒代蛋氨酸,而蛋黄样品主要成分为硒代半胱氨酸,还有少量的硒代蛋氨酸,同时,一些鸡蛋还含有极少量的亚硒酸根,但蛋清及蛋黄中硒代氨基酸的含量总和占总硒含量的83.3%~98.4%,适用于富硒鸡蛋中的硒形态提取。由此可见,市场上富硒鸡蛋中的硒主要以硒代氨基酸的形式存在,比较适合日常补硒,建立的方法可以对市场上富硒食品起到一定监测作用。

Selenium speciation determines the angiogenesis effect through regulating selenoproteins to trigger ROS-mediated cell apoptosis and cell cycle arrest

Tumor angiogenesis is closely related to tumor development,immune escape,and drug resistance.Therefore,the development of effective anti-tumor angiogenesis drugs is of great research significance.Although the current clinical angiogenesis inhibitors have achieved certain efficacy,they also pose the problems of limited and short duration of efficacy,drug resistance,and intrinsic toxicity.Anti-tumor angiogenesis strategies targeting endothelial cells(ECs)have attracted widespread attention in the development of highly effective and low toxicity anti-angiogenesis inhibitors.Studies have verified that the trace element selenium(Se)can inhibit tumor growth by inhibiting tumor angiogenesis through different mechanisms.Nevertheless,it is unclear whether Se speciation has different effects on anti-tumor angiogenesis.Herein,we found that Se exhibited effective anti-angiogenic activity,and its mechanisms of activity were determined by its chemical speciation.Organic Se can significantly inhibit tumor angiogenesis by targeting thioredoxin reductase(TrxR)to trigger cell apoptosis and cell cycle arrest and by increasing reactive oxygen species(ROS)production in ECs.Inorganic Se can induce cell cycle arrest and increase ROS production in ECs,showing promising anti-angiogenic effects.Se nanoparticles(SeNPs)slightly inhibit tumor angiogenesis by inducing apoptosis and cell cycle arrest and by increasing the production of ROS.In summary,this study elucidates the anti-angiogenic activity of Se speciation control with a view to providing a scientific reference for the design and development of novel Se-based highly effective and low toxicity angiogenesis inhibitors.

表生环境中硒形态研究现状

硒是一种对环境具有灵敏指示意义的元素,在表生环境中存在诸多形态,如Se(IV)、Se(VI)、Se(0)、Se(-II)、Org-Se,每种形态都有独特的化学、生物化学性质(控制元素的可溶性、毒性、营养功能、环境行为)。在综合分析近年来国内外表生环境硒形态研究的基础上,作者分别对岩石(矿物)、土壤和沉积物、生物体、各种水体、大气中硒自然形态的分布进行了综述,并初步建立了不同体系之间硒在积累、迁移过程中形态迁移转化的地球化学模型;最后指出了硒形态研究方面有可能的前景。

外源四价硒条件下硫对小麦硒吸收的影响机制

【目的】研究硫肥对外源四价硒条件下土壤硒赋存形态和价态变化的影响,旨在揭示硫调控小麦硒吸收的作用机制。【方法】以郑麦9023为试验材料,通过苗期土培和水培进行,土培试验设3个硫水平,分别为0、150和300 mg·kg-1,硫源为硫磺,2个硒水平,分别为0和5 mg·kg-1,硒源为亚硒酸钠,生长至70 d收获并取土样;水培试验先将幼苗在1/5 Hogland-Arnon营养液中培养2周,然后开始进行处理,设3个硫水平,分别为0、1和2 mmol·L-1,硫源为硫酸镁,硒水平为10μmol·L-1,2种硒源,分别为亚硒酸钠和硒酸钠,培养24 h后收获,土培和水培试验均为完全交互设计,重复4次,采用HG-AFS-8220型双道原子荧光光度计对小麦地上部和根以及土壤中硒含量进行测定,根据与植物硒吸收的相关性,将土壤总硒分成不同赋存形态和价态。【结果】施用适量硒和硫对小麦生长均有一定的促进作用;150 mg·kg-1硫可以显著降低小麦地上部和根中硒含量和硒累积量,降幅分别可达61.7%、34.4%和55.7%、24.7%,但是这种降低不会随施硫量的增加而显著变化;施硫可以显著降低土壤p H值,最高可降低0.50个单位,并增加了土壤有机质含量,最高可增加0.78 g·kg-1。施硫可以显著降低土壤中水溶态硒含量,并显著增加铁锰氧化物结合态硒含量,也增加了有机结合态和残渣态硒含量,而对交换态硒含量无显著影响,表明施硫促使了硒在土壤中的钝化。施硫可以显著降低水溶态中各种价态硒含量,其降幅基本一致,施硫可以增加交换态中四价硒含量而显著减少六价硒含量,表明施硫抑制了硒在土壤中向高效价态的转化。小麦对四价硒和六价硒的吸收差异与是否施硫密切相关,无硫时,六价硒处理下小麦地上部和根中硒含量分别为四价硒处理的44.7和22.4倍,而硫浓度为1 mmol·L-1时,其硒含量迅速降为四价硒处理的2.8倍和51.8%,表明硫可以显著缩小小麦对四价和六价硒的吸收能力差异。【结论】施用适量硫肥在改善小麦生长的同时,可以通过降低土壤p H和提高有机质含量,促使水溶态硒在土壤中向铁锰氧化物结合态、有机结合态及残渣态的钝化,并抑制交换态中四价硒向六价态的转化,从而降低小麦对硒的吸收。因此,可以通过在高硒缺硫地区增施硫肥以及缺硒高硫地区减施硫肥来有效调控作物体内的硒含量。

高效液相色谱-氢化物发生-原子荧光光谱法测定富硒食品中5种形态硒的含量

提出了将高效液相色谱-氢化物发生-原子荧光光谱法应用于富硒食品中硒的形态分析。选择木瓜蛋白酶(PP)作为提取剂,在如下优化条件下对样品中5种形态硒进行提取:(1)料液比1g/40mL;(2)50.0g·L^(-1) PP溶液加入量20mL;(3)于60℃水浴中振荡提取8h。提取液冷却并离心后取其上清液,用Hamilton PRP-X100离子交换色谱柱作为固定相,35mmol·L^(-1)磷酸氢二铵溶液(pH 6.0)作为流动相进行高效液相色谱的分离,5种形态硒[Se(Ⅵ)、Se(Ⅳ)、硒代胱氨酸(SeCys_2)、甲基-硒代半胱氨酸(MeSeCys)和硒代蛋氨酸(SeMet)]在16 min内完全分离。将分离所得分别含有5种形态硒的溶液先后引入仪器进行氢化物发生-原子荧光光谱法测定。结果表明:5种形态硒的质量浓度均在5~100μg·L^(-1)内与其对应的峰面积呈线性关系,5种形态硒的检出限(3S/N)为0.39~1.18μg·L^(-1)。按所提出方法对富硒大米样品进行加标回收和精密度试验,测得回收率为80.2%~118%,测定值的相对标准偏差(n=6)为2.1%~8.2%。

高效液相色谱-电感耦合等离子体质谱法检测富硒食品中6种硒形态

建立了高效液相色谱-电感耦合等离子体质谱联用技术(HPLC-ICP-MS)测定富硒食品中6种硒形态的方法。采用Hamilton PRP-X100分析柱,在流动相为20 mmol/L柠檬酸,pH=6.0,梯度洗脱的条件下,15 min内可以完全分离6种硒形态。所建立的方法简便、快速、灵敏度高,在色谱进样量为100μL,质谱采用He碰撞池模式下,对SeO2!4,SeO2!3,SeMet,SeCys2,SeUr,SeEt的检出限分别为0.7,0.5,4.1,1.3,13.6和11.9μg/L。利用含SeMet的富硒玉米粉探讨了不同提取方法的提取效果,并应用该方法测定了富硒大米、富硒酵母、富硒茶叶、富硒灵芝孢子粉等多种含硒功能食品中的硒形态,分析结果令人满意。

改良剂与生物有机肥配施方式对强酸性高硒茶园土壤硒有效性的影响

通过田间试验,研究了3种碱性改良剂(秸秆炭、生石灰粉和钙镁磷肥)与"爸爱我"生物有机肥的不同配施方式对强酸性高硒茶园土壤硒有效性的影响。结果表明:在无外源硒素添加的前提下,通过这种内源调控途径能够有效提高强酸性高硒茶园土壤硒的生物有效性。秸秆炭、生石灰粉和钙镁磷肥与"爸爱我"生物有机肥的3个错时配施处理对提高土壤硒有效性的效果最佳,分别使土壤有效硒含量提高105.98、114.09和146.71μg/kg,茶叶中硒含量相应提高0.96、1.14和1.24μg/g,增幅均达到极显著水平(P<0.01)。

基于路径分析的土壤性质与硒形态的关系

选择我国15种不同类型土壤,采用连续浸提法测定了土壤中硒的形态,并以Amos18.0路径分析法研究了土壤理化性质对土壤硒形态的影响。结果表明,除江西红壤(水田和旱田)和黑龙江黑土中硒主要是以残渣态存在外,其余土壤中硒主要以有机结合态为主,可溶态硒仅占土壤硒总量的0.4%～14.6%。土壤可交换态及碳酸盐结合态、有机结合态和残渣态硒含量间呈显著正相关(p<0.01),而可溶态和铁锰氧化物结合态硒含量与其他形态硒含量间无显著相关。路径分析表明,土壤有机质含量对铁锰氧化物结合态硒有负影响作用,而对其他四形态硒均为正影响作用。无定形铁含量对可溶态硒含量无影响作用,但对其他四形态硒的正影响作用大于有机质含量的。除可溶态硒外,土壤有机质和无定形铁含量分别对有机结合态和铁锰氧化物结合态硒的影响最大。pH和黏粒分别通过对土壤有机质和无定形铁的负和正作用而影响硒形态。有机质和无定形铁对硒在土壤中的分配起直接决定作用,而pH和黏粒也是不可忽略的影响因素。

硒减轻油菜幼苗硼毒害机理的研究

【目的】硼是植物生长发育所必需的微量元素,但在土壤中过量存在会对植物产生毒害。在干旱半干旱地区硼毒害是限制作物生长的重要因子,此外,硼矿开采以及硼肥用量过大或施用不均匀均会导致硼浓度过高,从而限制作物生长。适量硒能提高作物抗性,减轻作物因逆境产生的伤害,本文旨在通过适量硒处理,研究硒对高硼条件下油菜生物量及油菜活性氧代谢的变化,探讨硒缓解高硼对油菜生长抑制作用的机理,进而提高作物对硼的适宜范围,减轻硼毒害对农业生产的危害。【方法】本文以油菜(湘农油571)为试验材料,设置硼和硒2因素2水平(硼水平为:50、500μmol·L-1,分别用B50、B500表示,硒水平为:0、1μmol·L-1,分别用Se0、Se1表示)完全随机试验,共4个处理,通过苗期溶液培养,研究硒对高硼(500μmol·L-1)胁迫条件下油菜幼苗生长、硼吸收累积和抗氧化酶及非酶系统的影响。【结果】高硼胁迫条件下油菜幼苗地上部和地下部干物质重显著降低,与正常硼(50μmol·L-1)处理相比分别降低20.1%和32.0%;硼含量和累积量显著增加,与正常硼处理相比分别增加2.95和2.97倍;油菜幼苗因硼毒害导致叶片内抗氧化酶(过氧化氢酶(CAT)、过氧化物酶(POD)和抗坏血酸过氧化物酶(APX))活性和非酶抗氧化物(谷胱甘肽(GSH))含量显著降低,与正常硼处理相比,CAT、POD和APX活性分别降低19.7%、11.0%和15.0%;而过氧化氢(H2O2)和丙二醛(MDA)含量显著增加,比正常硼处理分别增加19.0%和18.5%。高硼条件下,经1μmol·L-1硒处理,与无硒处理相比,地上部和地下部干物质重分别增加22.8%和28.6%,硼含量和累积量分别降低38.6%和31.9%;油菜幼苗叶片内抗氧化酶活性和非酶抗氧化物含量显著增加,CAT、POD和APX活性分别增加12.8%、15.1%和15.3%,GSH和ASA含量分别增加9.7%和21.0%;而过氧化氢和丙二醛含量分别降低25.1%和21.2%。【结论】适量硒可降低高硼胁迫下油菜幼苗硼的吸收累积,增强抗氧化酶和非酶系统抗氧化作用,降低活性氧累积,减轻细胞质膜因过氧化作用而产生的损伤,从而显著减轻过量硼对油菜生长的影响,提高油菜幼苗对硼过量的适应性。

不同形态硒向水稻籽粒转运途径及品种差异

硒是人体必需微量元素,提高水稻籽粒硒含量对改善人体膳食硒营养有重要意义。以富硒水稻品种(Oryza sativa L.)秀水48和非富硒品种S.Andrea为材料,在灌浆期分别供应离体穗亚硒酸盐、硒酸盐、硒代蛋氨酸(SeMet)和硒甲基硒代半胱氨酸(SeMeSeCys),探讨两品种水稻在灌浆期向籽粒转运不同形态硒的品种差异及转运途径。结果表明:水稻体内有机硒主要通过韧皮部转运至籽粒,硒酸钠可能通过木质部和韧皮部共同转运至剑叶,而亚硒酸钠主要通过木质部转运至剑叶。秀水48从茎至籽粒转运硒酸盐和硒代蛋氨酸能力显著强于S.Andrea,并且富硒水稻秀水48从剑叶至籽粒转运有机硒(硒代蛋氨酸)能力显著高于S.Andrea。与非富硒水稻相比较,富硒水稻能通过茎和剑叶向籽粒转运较多的硒,这可能是引起水稻籽粒硒含量差异的直接原因。

硒酵母中硒的化学形态分析与评价

利用Ｔｒｉｓ－缓冲液－甘油提取法对硒酵母的化学形态进行了分析，并与透析法进行比较。结果表明，经过特殊工艺生产的硒酵母虽然总硒含量达到４０００μｇ／ｇ以上，但其中５０％以上的硒为硒蛋氨酸形态的蛋白质或其它大分子态硒，无机态硒仅占３％左右，而游离态硒只占０．０１％。说明硒酵母有广阔的开发前景，同时也为硒制剂提供了适宜的质量监测方法。

液相色谱-氢化物发生原子荧光光谱法测定富硒酵母中硒的形态

建立了富硒酵母中硒酸Se（Ⅵ）、亚硒酸Se（Ⅵ）、硒代蛋氨酸（SeMet）和硒甲基硒代半胱氨酸（SeMeCys）4种硒形态的高效液相色谱-氢化物发生原子荧光光谱（HPLC-HG-AFS）分析方法。样品采用蛋白酶和胰蛋白酶酶解提取,20 mmol/L（NH4）2HPO4为流动相,经PRP-X100阴离子交换色谱柱分离后HG-AFS测定。结果显示,4种硒形态标准曲线的线性关系良好（R^2≥0.9995）,方法检出限为0.5~5.0μg/kg,平均回收率为82.5%~101.2%,日内相对标准偏差≤8.6%,日间相对标准偏差≤14.5%。本方法前处理简单、灵敏度高、设备便宜、运行费用低廉,适用于富硒饲料产品中硒的形态分析。

硒对高温胁迫下辣椒叶片抗氧化酶活性的调节作用

硒是人体或动物抗氧化反应必需的微量元素。为了研究硒在高等植物体内的生理功能,将辣 椒两个品种‘中椒7号’和‘耐湿椒’盆栽在不同含硒量的草炭蛭石混合基质中,现蕾后分次将植株移入 人工气候室进行高温胁迫处理(25℃2h→35℃2h→40℃2h),然后取叶片测定各指标。结果表明:施硒 (≤0.9mg·kg-1)提高了叶片的总叶绿素含量,同时,可以分别使GPX和POD活性提高11.5倍和 63.6%,对SOD和CAT则表现为负效应。另外,硒还使叶片MDA含量比不加硒对照降低36%。在不同品 种的辣椒体内,硒的作用存在一些差异,但总体的趋势相同。以上结果说明硒对高温胁迫下辣椒叶片抗氧 化酶活性有重要的调节作用,并有利于提高辣椒植株的耐热性。

高效液相色谱-氢化物发生-原子荧光光谱联用技术测定富硒大米中的5种硒形态

目的建立高效液相色谱-氢化物发生-原子荧光光谱法(high performance liquid chromatographyhydride generation-atomic fluorescence spectrometry,HPLC-HG-AFS)检测富硒大米中硒代胱氨酸(Se Cys2)、甲基硒代半胱氨酸(Me Se Cys)、亚硒酸根(Se IV)、硒代蛋氨酸(Se Met)和硒酸根(Se VI)的方法。方法选择Hamilton PRP-X100阴离子交换色谱柱,对流动相浓度、p H值以及提取条件进行选择优化。样品采用链霉蛋白酶E水解超声提取方式,使用40 mmol/L的磷酸氢二铵溶液(p H 6.0)作为流动相。结果 20 min内可以完全分离5种硒形态,各种硒形态标准曲线线性相关系数(r)均大于0.999,检出限分别为3.7、1.7、1.2、4.2、7.2μg/L。加标回收实验显示富硒大米中硒的主要存在形态即硒代蛋氨酸回收率范围在95.1%~98.2%,其他4种硒形态回收率范围均在80.4%~102.5%。结论本方法可以快速、准确地测定大米中硒的5种形态。