硒酵母中硒含量测定方法的研究

报道了人工培养硒酵母中总硒、无机硒和有机硒的测定方法,建立了无机硒和有机硒的鉴别方法,并采用透析处理法使硒酵母中的无机硒和有机硒得以分离;还报道了不同培养条件下得到的5种硒酵母中总硒、无机硒和有机硒的定量分析数据,为硒酵母在医药和添加剂领域的应用提供了可靠的数据。     

大米中硒含量的测定

为使人们更好地控制和调节硒的摄入量,用紫外分光光度法测定了大米中的硒含量。样品通过混酸消化,将有机硒转化成无机硒,酸性条件下,Se(Ⅳ)与邻苯二胺反应,产物在335nm有最大吸收峰。测定结果表明,天津小站米、东北米、丝苗米、泰国香米、富硒米的有机硒含量分别为:0.041、0.046、0.035、0.034、2.021μg/g,有机硒含量占总硒含量的比例分别是89.13%、82.14%、87.50%、80.95%、98.92%。富硒米硒含量及有机硒的比例明显高于其它种类的米。     

控温消解-氢化物发生-原子荧光光谱法测定水中硒形态

取10.00 mL水样,于60～70℃加热蒸发至近干,加入硝酸-高氯酸(1+1)混合酸2.0mL,继续加热至白烟冒尽,加入盐酸2.0 mL,摇匀,加热保持微沸3～5 min,冷却后转移至10mL比色管中,用去离子水定容,使用氢化物发生-原子荧光光谱仪测定总硒的含量。另取10.00mL水样,加入盐酸2.0mL,于60～70℃加热至溶液体积小于5mL,转移至10mL比色管中,用去离子水定容,使用氢化物发生-原子荧光光谱仪测定无机硒(即四价硒和六价硒的总和)的含量。另取水样5.00mL于10mL比色管中,加入盐酸2.0mL,用去离子水定容,使用氢化物发生-原子荧光光谱仪测定四价硒的含量。用总硒含量减去无机硒含量即为有机硒含量,无机硒含量减去四价硒含量即为六价硒含量。在最佳仪器工作条件下,硒的质量浓度在1.00～20.0μg·L~(-1)内与其对应的荧光强度呈线性关系,硒的检出限(3s)和测定下限(10s)分别为0.11,0.36μg·L~(-1)。采用本方法测定某地区水中的总硒、无机硒和四价硒,加标回收率在95.7%～104%之间,相对标准偏差(n=6)在1.9%～3.2%之间。采用本方法测定了不同地区水中的总硒、无机硒和四价硒,利用差减法计算得有机硒和六价硒的含量。     

硒酵母中有机硒及硒代氨基酸含量的测定方法

报道了人工培养硒酵母中有机硒及硒代胱氨酸（ＳｅＣｙｓ）和硒代蛋氨酸（ＳｅＭｅｔ）含量的测定方法。采用透析处理法使硒酵母中的无机硒和有机硒得以分离,并采用催化分光光度法测定了硒酵母中有机硒的含量;采用氨基酸自动分析仪测定了硒酵母中ＳｅＣｙｓ和ＳｅＭｅｔ的含量。     

顺序注射-氢化物发生-原子荧光光谱法测定海产品中无机硒和有机硒

海产品样品(2.500 0g)采用盐酸(1+1)溶液25mL于60℃提取18h,经脱脂棉过滤,滤液经环己烷5mL萃取后分层;取水相4.00mL采用顺序注射-氢化物发生-原子荧光光谱法测定其中无机硒的含量。另取样按微波消解-原子荧光光谱法测定此样品中的总硒量。采用差减法计算有机硒的含量。无机硒的质量浓度在1.00~20.0μg·L~(-1)范围内与其荧光强度呈线性关系,检出限(3s)为0.113μg·L~(-1)。在0.100,1.00,2.00mg·kg~(-1)等3个浓度水平进行加标回收试验和精密度试验,回收率在84.8%~93.5%之间,测定值的相对标准偏差(n=6)在1.6%~2.5%之间。     

反相离子对高效液相色谱-电感耦合等离子体质谱法测定富硒酵母中硒

富硒酵母样品用pH 7.5的Tris-盐酸缓冲溶液经超声破碎提取30min,于提取液中加链霉蛋白酶E在37℃搅拌18h,高速离心分离。取上清液经Millipore Corp超滤膜过滤,滤液中无机硒(Ⅵ及Ⅳ)和有机硒,即硒代半胱氨酸、硒代尿素和硒代蛋氨酸通过Waters Symmetry ShieldRP18色谱柱分离。用不同比例混合的甲醇-七氟丁酸溶液作流动相进行梯度洗脱,用电感耦合等离子体质谱法在线测定洗脱液中各种形态硒的含量。5种形态硒的质量浓度均在100μg·L-1以内呈线性。应用此法分析了富硒酵母样品,结果表明:无机硒占总硒量的1.63%,65%的硒以易被人体吸收的硒代蛋氨酸形式存在。     

乙酸乙酯萃取-氢化物发生-原子荧光光谱法测定高纯金中硒和碲

硒和碲是高纯金中的主要有害杂质元素,依据国家标准GB/T 25933-2010规定高纯金中硒和碲是必检项目,所用仪器为电感耦合等离子体原子发射光谱仪(ICP-AES)[1-2]和电感耦合等离子体质谱仪(ICP-MS)[3],而这两种仪器成本和运行费用十分昂贵,且检出限较高。氢化物发生-原子荧光光谱法(AFS)具有灵敏度高、检出限低、仪器相对便宜的优点,已广泛应用于冶金、地质和生物样品分析[4-5]。     

高效液相色谱-电感耦合等离子体质谱分析几种富硒产品的硒总量及其形态

本文采用高效液相色谱-电感耦合等离子体质谱(HPLC-ICP-MS)分析了几种富硒产品中硒的总量及其形态。结果表明,四种富硒产品的硒总量均有不同程度增加,富硒大米和富硒茶叶中的硒总量较高,分别为1.300μg/g和0.459μg/g,富硒大米和富硒大葱的富硒倍率相对较高,分别为未富硒产品的24.1和17.8倍。以0.1mol/LHCl辅助超声预处理样品进行富硒螺旋藻和富硒大葱中硒的形态分析,发现富硒大葱中的硒主要为有机态的甲基硒代半胱氨酸(MeSeCys)以及部分硒代蛋氨酸(SeMet),而富硒螺旋藻中的硒大部分为无机硒。说明经根部吸收等富集方式可有效将无机硒转化为有机硒,而某些方式未能完全转化。     

两种干海产品中硒含量的紫外-可见分光光度间接测定

基于Se(IV)与I-的反应建立了紫外-可见分光光度法间接测定干海产品中总硒及无机硒的含量。获得影响分析性能的最佳因素:最大吸收波长516 nm,反应体系中2%KI、6.0 mol·L~(-1)HCl及0.02 mol·L~(-1)EDTA-2Na的加入量分别为0.6 m L、1.0 m L、2.0 m L,室温下反应25 min。25 m L反应液中硒含量(μg)在0.05~14.4范围内与吸光度呈线性:A=-0.0049+0.05176c,r=0.9998,检测限0.01μg/25 m L(3S/k)。成功用于干海产品虾皮及银鱼中总硒及无机硒含量测试。结果表明有机硒含量占总硒67%以上,总硒及无机硒的回收率分别为95.3~99.3%、101.8~108.1%。该法操作简单,抗干扰能力强,具有良好准确度及重现性。     

氢化物发生-原子吸收光谱法测定黄芪中硒

黄芪试样经硝酸-过氧化氢消解,用氢化物发生-原子吸收光谱法测定黄芪中总硒、无机硒和有机硒的含量。使用15 g·L^-1硼氢化钾-3 g·L^-1氢氧化钠溶液与溶液中硒离子反应生成氢化物。分析中采用载气流量240 mL·min^-1。硒的质量浓度在60μg·L^-1以内与吸光度呈线性关系,方法的检出限（3σ）为0.21μg·L^-1。应用此法测定黄芪中硒的含量,总硒测定值为105μg·g^-1、硒（Ⅳ）测定值为15μg·g^-1、硒（Ⅵ）测定值为9μg·g^-1、有机硒测定值为81μg·g^-1。黄芪中的硒主要存在形式为有机硒,占总硒含量的77%。     

Quantitative analysis of volatile selenium metabolites in normal urine by headspace solid phase microextraction gas chromatography-inductively coupled plasma mass spectrometry

The combination of headspace-solid phase microextraction (HS-SPME) and gas chromatography-inductively coupled plasma mass spectrometry (GC-ICPMS) was evaluated for the determination of volatile selenium metabolites in normal urine samples, i.e. without selenium supplementation. HS-SPME operating conditions were optimised and a sampling time of 10 min was found to be suitable for simultaneous extraction of dimethylselenide (DMSe) and dimethyldiselenide (DMDSe). The amount of DMSe and DMDSe extracted onto fibre coating was calculated in clean matrix, i.e. Milli-Q water, on the basis of depletion experiments. When applied to normal urine samples, the developed method allowed the detection of four volatile selenium containing species, among which DMSe and DMDSe could be quantified by standard additions.     

Pervaporation as a Separation Technique for Direct Determination of Volatile Selenium Species by Atomic Fluorescence Spectrometry

Abstract

This paper reports for the first time a suitable way to determine methylated selenium compounds using the new approach of pervaporation coupled to atomic fluorescence spectrcmetry (PV-AFS).

The method developed allows direct extraction, separation, preconcentration and determination of dimethylselenium (DMSe) and dimethyldiselenium (DMDSe) from slurry samples. Under the optimum conditions, the detection limits (LODs) were found to be 0.66 ng and 0.39 ng for DMSe and DMDSe, respectively, the precision being about 6–9 % for 10 ng mL as selenium concentration. The linearity ranges were from the LOD to 0.7 μg mL^?1 for DMSe and from the LOD to 0.4 μg mL^?1 for DMDSe (as Se). The pervaporation efficiencies were 55 ± 1 % and 85 ± 5 % for DMSe and DMDSe, respectively. The proposed method was successfully applied to determine methylated selenium species in sewage sludge, garlic and oyster samples. The concentrations found were from 0.07 to 1.42 μg g^?1.

As no certified reference materials are available for these analytes, validation was carried out by recovery studies in these matrices, and the results showed that the proposed method performed satisfactorily.     

Headspace hollow fiber protected liquid-phase microextraction combined with gas chromatography-mass spectroscopy for speciation and determination of volatile organic compounds of selenium in environmental and biological samples

A simple and novel speciation method for the determination of volatile organic compounds of selenium (dimethylselenide (DMSe) and dimethyldiselenide (DMDSe) has been developed using a headspace hollow fiber protected liquid-phase microextraction (HS-HF-LPME) combined with capillary gas chromatography-mass spectrometry (GC-MS). The organic solvent impregnated in the pores and filled inside the porous hollow fiber membrane was used as an extraction interface in the HS-HF-LPME of the compounds. The effect of different variables on the extraction efficiency was studied simultaneously using an experimental design. The variables of interest in the HS-HF-LPME were sample volume, extraction time, temperature of sample solution, ionic strength, stirring rate and dwelling time. A Plackett-Burman design was performed for screening in order to determine the significant variables affecting the extraction efficiency. Then, the significant factors were optimized by a Box-Behnken design (BBD) and the response surface equations were derived. Under optimum conditions, preconcentration factors up to 1250 and 1170 were achieved for DMSe and DMDSe respectively. The detection limit and relative standard deviation (RSD) (n=5, c=50 μg L(-1)) for DMSe were 65 ng L(-1) and 4.8%, respectively. They were also obtained for DMDSe as 57 ng L(-1) and 3.9%, respectively. The developed technique was found to be applicable to spiked environmental and biological samples.     

On-line organoselenium interference removal for inorganic selenium species by flow injection coprecipitation preconcentration coupled with hydride generation atomic fluorescence spectrometry

The existence of dimethylselenium (DMSe) and dimethyldiselenium (DMDSe) in some environmental samples can cause serious interference on Se(IV) determination by hydride generation atomic fluorescence spectrometry (HG-AFS) due to their contribution on HG-response. A flow injection separation and preconcentration system coupled to HG-AFS was therefore developed by on-line coprecipitation in a knotted reactor (KR) for eliminating interference subjected from organoselenium. The sample, spiked with lanthanum nitrate, was merged with an ammonium buffer solution (pH 8.8), which promoted coprecipitation of Se(IV) and quantitative collection by 150 cm PTFE KR. DMSe and DMDSe, however, were unretained and expelled from the KR. An air flow was introduced to remove the residual solution from the KR, then a 1.2 mol l⁻¹ HCl was pumped to dissolve the precipitates and merge with KBH₄ solution for HG-AFS detection. The interference of DMSe and DMDSe on the Se(IV) determination by conventional HG-AFS and its elimination by the developed separation and preconcentration system were evaluated. With optimal experimental conditions and with a sample consumption of 12.0 ml, an enhancement factor of 18 was obtained at a sample frequency of 24 h⁻¹. The limit of detection was 0.014 μg l⁻¹ and the precision (R.S.D.) for 11 replicate measurements of 1.0 μg l⁻¹ Se(IV) was 2.5%. The developed method was successfully applied to the determination of inorganic selenium species in a variety of natural water samples.     

Gas chromatography with atomic emission detection for dimethylselenide and dimethyldiselenide determination in waters and plant materials using a purge-and-trap preconcentration system

Dimethylselenide (DMSe) and dimethyldiselenide (DMDSe) were determined in plant and water samples by capillary gas chromatography using microwave induced-plasma atomic emission spectrometry for detection. The analytes were leached from the solid samples into methanol by using an ultrasonic probe, and a portion of the extract was preconcentrated by means of a purge-and-trap system before being chromatographed. The analytes were directly purged from the water samples in the presence of 6% (v/v) methanol. Element-specific detection and quantification was carried out by monitoring the selenium (196 nm) emission line. Calibration curves were obtained by plotting peak area versus concentration and the correlation coefficients for linear calibration were 0.9999 for both analytes. Detection limits of 0.8 and 1.1 ng l(-1) were obtained for DMSe and DMDSe, respectively, for water samples. For plant materials, the detection limits calculated for 0.5 g samples were 0.3 and 0.4 ng g(-1) for DMSe and DMDSe, respectively. Concentration levels of DMSe ranging from 1.2 to 4.2 ng g(-1) were found in some of the plant materials analyzed. No DMDSe was found in any of the samples. The accuracy of the method was checked by analyzing different spiked water and plant samples.     

The effect of the presence of volatile organoselenium compounds on the determination of inorganic selenium by hydride generation

As a result of microbiological activity it is possible to find dimethylselenium (DMSe) and dimethyldiselenium (DMDSe) in a wide type of environmental samples, such as soils, sediments, sewage sludges and plants where methylation can take place.Selenium determination by hydride-generation (HG) techniques requires its presence as Se(IV). Consequently, inorganic speciation by hydride generation techniques is done by first determining Se(IV) and then, after reduction of Se (VI) to Se(IV), the total selenium. Therefore, the concentration of Se (VI) is evaluated as the difference between total inorganic selenium and Se(IV). In the present work it could be demonstrated that DMSe and DMDSe are forming other volatile species by reaction with sodium borohydride, applying the same reduction condition as for inorganic selenium. These species are subsequently detected by several atomic techniques (atomic absorption AAS, atomic fluorescence AFS and inductively coupled plasma-mass spectrometry ICP-MS). The error that their presence can cause in determination of inorganic selenium has been evaluated. The magnitude of this error depends on the specific analytical detector used.The coupling of pervaporation-atomic fluorescence is proposed for the identification of these species and pervaporation-gas chromatography-atomic fluorescence for their individual quantification.     

Determination of dimethylselenide and dimethyldiselenide in milk and milk by-products by solid-phase microextraction and gas chromatography with atomic emission detection

A method based on solid-phase microextraction (SPME) followed by gas chromatography with microwave-induced plasma atomic emission detection for determining dimethylselenide (DMSe) and dimethyldiselenide (DMDSe) in milk and milk by-products is proposed. Parameters affecting the SPME, such as sample volume or mass, ionic strength, adsorption and desorption times and temperatures were optimized in the headspace mode. The matrix effect was evaluated for the different samples studied, concluding that standard additions calibration was required for quantification purposes. The detection limits ranged from 70 to 110 pg mL⁻¹ for DMSe and from 80 to 400 pg mL⁻¹ for DMDSe, depending on the sample under analysis. None of the twenty-three samples analyzed contained the studied compounds at concentrations above the corresponding detection limits.     

An Organodiselenide Comediator to Facilitate Sulfur Redox Kinetics in Lithium–Sulfur Batteries with Encapsulating Lithium Polysulfide Electrolyte

Lithium–sulfur (Li–S) batteries are regarded as promising high-energy-density energy storage devices. However, the cycling stability of Li–S batteries is restricted by the parasitic reactions between Li metal anodes and soluble lithium polysulfides (LiPSs). Encapsulating LiPS electrolyte (EPSE) can efficiently suppress the parasitic reactions but inevitably sacrifices the cathode sulfur redox kinetics. To address the above dilemma, a redox comediation strategy for EPSE is proposed to realize high-energy-density and long-cycling Li–S batteries. Concretely, dimethyl diselenide (DMDSe) is employed as an efficient redox comediator to facilitate the sulfur redox kinetics in Li–S batteries with EPSE. DMDSe enhances the liquid–liquid and liquid–solid conversion kinetics of LiPS in EPSE while maintains the ability to alleviate the anode parasitic reactions from LiPSs. Consequently, a Li–S pouch cell with a high energy density of 359 Wh kg⁻¹ at cell level and stable 37 cycles is realized. This work provides an effective redox comediation strategy for EPSE to simultaneously achieve high energy density and long cycling stability in Li–S batteries and inspires rational integration of multi-strategies for practical working batteries.     

Comparison of two derivatizing agents for the simultaneous determination of selenite and organoselenium species by gas chromatography and atomic emission detection after preconcentration using solid-phase microextraction

Two methods for the simultaneous determination of selenite and two organoselenium compounds, dimethylselenide (DMSe) and dimethyldiselenide (DMDSe), are proposed. Both methods involve sample preconcentration by solid-phase microextraction (SPME) and capillary gas chromatography coupled to atomic emission detection (GC-AED). The main difference between the methods is the derivatizing agent used to complex the inorganic species: sodium tetraethylborate and 4,5-dichloro-1,2-phenylenediamine. The parameters affecting the derivatization and preconcentration steps, chromatographic separation as well as detection of the compounds were optimized. Direct immersion (DI) mode and a relatively long extraction time were selected for the method involving the formation of the piazselenol complex, better sensitivity being achieved for the three analytes under study. In this case, detection limits ranged between 3 and 25 ng L(-1), depending on the compound. Headspace mode (HS) and extraction times of 20 min were selected for the method involving tetraalkylborate, and detection limits of between 7.3 and 55 ng L(-1) were obtained. DMSe and Se(IV) were found in several of the water samples analyzed at concentrations of 0.07-1.0 ng mL(-1).     

Formation of volatile selenium species in synthetic seawater under light and dark experimental conditions

Experiments were performed in the laboratory on synthetic seawater spiked with different selenium species at trace levels to study the formation of volatile selenium compounds under dark or controlled simulated sunlight conditions. Spiking the reaction media with inorganic and organic selenium compounds demonstrated that several volatile selenium species could be formed under these simulated conditions. Selenoamino acids react to produce significant amounts of volatile selenium species in both light and dark conditions. Products formed include dimethyl selenide (DMSe), dimethyl selenyl sulphide (DMSeS) and dimethyl diselenide (DMDSe). Inorganic selenium oxyanions added to the synthetic reaction media did not form volatile species via abiotic reactions despite the presence of strong methylating agents. These results suggest that the formation of stable volatile species from bio‐organic selenium compounds can occur via abiotic reactions in the marine photic zone. Copyright © 2000 John Wiley & Sons, Ltd.