应用体外仿生模型分析海藻水煎液中微量金属的形态和生物可给性

将体外"半仿生消化"和"全仿生消化"法分别应用于海藻分析前处理,即模拟人体消化环境,加入消化液所含有机物和无机物,模拟海藻(海带和紫菜)水煎液在胃肠中的吸收和转运机制.鉴于消化管和血管间的生物膜是类脂质膜,以单层脂质体为细胞生物膜模型考察海藻水煎液中微量金属在单层脂质体-水体系中的分配行为.以单层脂质体结合态、水溶态界定水煎液中微量金属的形态; 以单层脂质体结合态含量评价微量金属的生物可给性; 比较单层脂质体结合态金属在胃肠中浓度,确定微量金属的主要吸收部位.消化酶对海藻中微量元素的形态和生物可给性影响明显.水煎液全仿生消化后,海带和紫菜中的Fe均主要在胃被吸收,海带中的Mn和Zn主要在肠被吸收;紫菜中的Mn和Zn在胃与肠均有吸收.     

脂质体的特异性识别与单颗粒电化学检测

脂质体载药系统已在许多疾病的治疗中发挥重要作用.体外构建并模拟脂质体与生物膜特异性识别相互作用,对于探索脂质体在生物体内的稳定性,开发新型高效和智能化药物递送系统具有重要的现实意义.本文通过分子组装技术构筑了稳定的磷脂囊泡-脂质体,利用微加工技术制备金微电极,然后在其表面进行亲合素修饰,借助电化学工作站检测脂质体单颗粒的碰撞行为,以此建立一种检测脂质体在体外和生物体内存在的技术平台.实验证明,在脂质体中引入生物素分子,通过与微电极表面的亲合素分子产生特异性识别,能显著增加单颗粒碰撞的频率和强度.这些定量的数据为研究脂质体与生物膜动态互作提供了十分有效和准确的检测手段,为验证脂质体载药体系在生物内的稳定性和释放速率建立了新方法.     

脂质体为模板仿生硅化固定葡萄糖氧化酶

将脂质体囊泡与仿生硅化技术相结合,模拟细胞纳微环境,实现以脂质体为模板仿生制备氧化硅固定葡萄糖氧化酶(GOx),建立性能稳定的固定化酶.扫描电镜分析显示,固定化GOx为球形纳米粒子,粒径分布在200nm左右,在优化反应条件下GOx回收率达到71.8%.由于载体的空间限制作用及其提供的较稳定微环境,固定化GOx表现出良好的热稳定性和pH稳定性,其对变性剂耐受性和操作稳定性等也得到明显提高.     

用同步辐射X-荧光定量测定电泳分离后蛋白条带内的微量元素

建立了同步辐射X荧-光(SRXRF)定量测定生物样品等电聚焦(IEF)分离后蛋白条带内的微量元素Fe、Cu和Zn的方法。用薄层聚丙烯酰胺凝胶分离人血红蛋白后,用SRXRF测定了各亚型条带内的金属含量,用加一定量金属的含蛋白聚丙烯酰胺凝胶做SRXRF定量测定蛋白条带内微量元素Fe、Cu和Zn的定量标准,校准曲线线性回归系数r在0~8μg/g范围内均大于0.99;检出限分别为2.43、1.12和0.96μg/g;测定蛋白条带内Fe和Zn的回收率分别为90.4%和115.7%。该联用技术可用于生物样品中微量元素的化学形态分析,同时给出蛋白质的微量元素组成和等电点等信息。     

体积排阻高效液相色谱-电感耦合等离子体质谱法测定海产贝类中镉的形态

运用体积排阻高效液相色谱-电感耦合等离子体质谱联用技术(SEC-HPLC-ICP-MS)分析了高镉积累扇贝和低镉积累菲律宾蛤仔中镉的存在形态,并结合体外全仿生消化技术,研究了在唾液、胃、肠无机物和有机物(含消化酶)作用下,扇贝和菲律宾蛤仔中镉的主要存在形态.结果发现:扇贝中Cd总量约为菲律宾蛤仔的10倍;在扇贝中检测到3种Cd形态:金属硫蛋白(MT)-Cd、谷胱甘肽(GSH) Cd和半胱氨酸(Cys)-Cd;在菲律宾蛤仔中检测到2种Cd形态:MT-Cd和GSH-Cd;以峰面积作参考进行比较,扇贝中MT-Cd和GSH-Cd含量分别约为菲律宾蛤仔的5 6和2.0倍.结合体外全仿生模型发现,在扇贝胃全仿生提取液中,检测到1种未知小分子有机镉形态(Cd-X),在扇贝肠全仿生提取液中检测到4种Cd形态,其中MT-Cd是主要形态;而在菲律宾蛤仔胃、肠全仿生提取液中均仅检测到1种未知小分子有机态镉(Cd-X).本实验证明贝类中的MT-Cd,GSH-Cd,Cys-Cd中络合的Cd在生物体胃肠消化液作用下会发生解离.     

纳米雄黄与脂质体仿生膜的相互作用研究

本工作以卵磷脂与胆固醇组成的磷脂小单层脂质体（small unilamelarvesicles,suv）作为仿生膜的简单模型,采用表面等离子共振技术（SPR）、荧光偏振、拉曼（Raman）光谱、核磁共振（NMR）及原子力显微镜（AFM）研究纳米雄黄与SUV仿生膜的相互作用,证实了磷脂是纳米雄黄作用的关键靶分子．随纳米雄黄结合,SUV仿生膜的相对粘度聃值增大,膜的流动性减小．Raman光谱数据计算表明,作用后膜的纵向有序性参数s。。及横向有序性参数Slat值增大,说明纳米雄黄的结合使磷脂膜的脂酰基链全反式构型比例上升,膜的流动性减小．由Raman光谱和引PNMR结果推测,磷脂极性头部是纳米雄黄与磷脂的主要结合位点。AFM实时观测,纳米雄黄通过在膜表面打“孔”或“洞”的方式,损坏磷脂膜．     

体外仿生法研究牡蛎和蛤仔体内微量金属元素的生物可给性

采用体外全仿生消解方法,模拟人体的消化道环境,对牡蛎和蛤仔样品进行仿生消解,分析牡蛎和蛤仔体内微量元素的生物可给性.以原子吸收光谱法测定胃、肠仿生提取液中微量元素的含量.结果表明,牡蛎胃提取液中Fe,Cu,Zn,Cd的含量明显高于肠提取液中的含量,蛤仔胃提取液中Fe,Cu的含量与肠提取液中的含量没有明显差异,而Zn,Cd的含量远高于其在肠提取液中的含量.经全仿生消解后,牡蛎体Fe,Cu,Zn,Cd的可给性分别为60.2％,83.6％,83.1％,76.8％;蛤仔Fe,Cu,Zn,Cd的可给性分别为46.3％,86.3％,85.3％,87.7％.本研究结果为进一步研究牡蛎和蛤仔对金属元素的累积和迁移、生物可利用性,Cd等有害元素对食用者的健康风险评估,提供技术支持和理论依据.     

金樱子茎根中必需微量元素的测定

为测定和分析金樱子茎、根中的微量元素种类及含量,采用酸式(HNO3-H2O2)微波消解法处理样品,原子吸收分光光度法测定了金樱子根茎中的微量元素,统计学处理数据。结果表明,在自采茎、根中各种元素含量(单位均为μg/g)分别为:Cu(1.713±0.156),(1.857±0.191);Zn(204.541±5.377),(432.209±4.874);Ca(2 596.875±68.337),(1 554.541±42.743);Mg(429.041±19.740),(663.750±23.867);Mn(18.604±0.285 5),(14.304±0.884);Cr(1.672±0.079),(3.561±0.079);Ni(0.896±0.068),(0.488±0.025);Fe(60.900±0.651),(70.221±1.328);Co(0.758±0.068),(0.662±0.057)。茎与根比较有显著差异的是Ca、Cr、Ni、Mg、Zn。结论:金樱子茎根含有丰富的必需微量元素,但不同部位之间某些微量元素的含量存在一定差异。     

氧化物在载体上的存在状态及其还原性能研究——NiO/γ-Al_2O_3和CuO/γ-Al_2O_3体系

本文用透射电子显微镜、选区电子衍射和X光衍射研究NiO/γ-Al_2O_3和CuO/γ-Al_2O_3体系,结果表明,氧化物在载体表面有单层分散和晶相两种状态。当氧化物含量低于某一阈值时,呈单层分散态,高于此阈值时出现晶相。氢还原研究表明,单层分散态比晶态难还原。晶相NiO在300—400℃还原,单层分散态则要450—530℃;晶相CuO在约180℃时还原,单层分散态则要240—400℃。无论是单层分散的或晶相的氧化物,还原得到的金属都是晶相,但单层分散的氧化物还原出来的金属晶粒较小。     

金属基仿生超滑表面制造及其应用

金属是人类社会赖以生存和发展的重要物质基础,是构成现代文明社会的支撑材料,增加金属材料的特殊性能,拓宽金属材料的应用范围,已经成为自然科学交叉研究的热点。受自然界启发,研究人员通过探究猪笼草口缘区的超滑行为,在微/纳米结构基底中注入低表面能润滑液,形成固液复合涂层,设计并制备出具有特殊润湿性的仿生超滑表面,显现出优异的自愈、防冰、防污、耐腐蚀、抗生物黏附、自清洁等性能。该方法为金属基体上设计构建仿生超滑表面,并实现其表面多功能性,进而为其在海洋防污、生物医疗、航空航天、制冷、工业生产等领域实现更广泛的应用提供了可能。本文从仿生超滑表面的设计原理、制备工艺、金属基体仿生超滑表面的应用以及未来发展趋势和挑战四个方面对金属基体仿生超滑表面的研究进展进行了综述。     

仿生提取-电感耦合等离子体-质谱法测定柏子养心片中可溶性重金属

国家食品药品监督管理总局的2014年国家药品抽验计划将柏子养心片定为抽验品种,本实验利用人工胃液仿生提取样品可溶性重金属,通过微波消解技术对提取液进行消解处理,建立了仿生提取-ICP-MS法测定18批柏子养心片中 Co, Cr, Cu, Ba, Cd, Mn, Ni, Pb, Hg, Sr, Zn 11种可溶性重金属的含量的分析方法。本方法各元素标准系列线性回归方程的相关系数为0.9989~1.0000,检出限为0.19~5.6μg/ L,方法的重复性 RSD 值均小于6.2%,精密度的 RSD 值均小于5.6%,加标回收率在87.7%~101.9%之间,满足检测要求。参照《药用植物及制剂进出口绿色行业标准》规定的重金属限量标准,18批样品中 Pb, Cd, Cu 均未超过标准,Hg 含量平均值为7.68 mg/ kg,超过标准值。仿生提取-ICP-MS 法的建立为柏子养心片的安全性研究提供参考依据。     

On-line preconcentration method for the determination of trace metals in water samples using a fully automated pretreatment system coupled with ICP-AES.

An automated on-line sample-preparation method using a computer-controlled pretreatment system (Auto-Pret AES system) coupled with ICP-AES was developed. In this work, an iminodiacetate chelating resin, packed in a mini-column and installed in the system was employed for the collection/concentration of 13 trace metals, including such toxic metals as Be, Cd, Cr, Cu and Pb. The limits of detection of the proposed method for trace metals were in the range of 0.001 (Be) -0.18 (Pb) ng mL-1. The enrichment factors for metal ions were about 19 times, when 5 mL of samples were used. The sample throughput was 11 h-1. The accuracy and the precision of the method were evaluated using river-water reference materials, SLRS-4 from NRCC, JSAC 0301-1 and JSAC 0302 from the Japan Society for Analytical Chemistry. The proposed method can be favorably applied to the collection/concentration of trace metals in natural water samples.     

On-line collection/concentration and determination of transition and rare-earth metals in water samples using Multi-Auto-Pret system coupled with inductively coupled plasma-atomic emission spectrometry

On-line preconcentration and determination of transition and rare-earth metals in water samples was performed using a Multi-Auto-Pret system coupled with inductively coupled plasma-atomic emission spectrometry (ICP-AES). The Multi-Auto-Pret AES system proposed here consists of three Auto-Pret systems with mini-columns that can be used for the preconcentration of trace metals sequentially or simultaneously, and can reduce analysis time to one-third and running cost of argon gas and labor. A newly synthesized chelating resin, ethylenediamine-N,N,N′-triacetate-type chitosan (EDTriA-type chitosan), was employed in the Multi-Auto-Pret system for the collection of trace metals prior to their measurement by ICP-AES. The proposed resin showed very good adsorption ability for transition and rare-earth metal ions without any interference from alkali and alkaline-earth metal ions in an acidic media. For the best result, pH 5 was adopted for the collection of metal ions. Only 5 mL of samples could be used for the determination of transition metals, while 20 mL of samples was necessary for the determination of rare-earth metals. Metal ions adsorbed on the resin were eluted using 1.5 M nitric acid, and were measured by ICP-AES. The proposed method was evaluated by the analysis of SLRS-4 river water reference materials for trace metals. Good agreement with certified and reference values was obtained for most of the metals examined; it indicates that the proposed method using the newly synthesized resin could be favorably used for the determination of transition and rare-earth metals in water samples by ICP-AES.     

Preconcentration of traces of silver and bismuth from cobalt and nickel and their nitrates, with activated carbon

A method is described for the preconcentration of trace metals Ag and Bi, present as impurities in high-purity cobalt and nickel metals and their nitrates. After the metal samples have been dissolved in nitric acid (or the salts in water) the trace elements are complexed with ammonium pyrrolidinedithiocarbamate (APDC). The sample solution is then filtered through a 2-cm filter paper coated with 50 mg of activated carbon, whereby the complexed trace metals are adsorbed on the activated carbon and separated from the matrix. The trace elements are dissolved off with nitric acid and determined by flame atomic-absorption spectrometry (AAS). The detection limits for the analysis of 10 g of metal samples and 50 g of the nitrate samples were 0.002-0.035 ppm Ag and 0.04 ppm Bi for the metal samples, and 0.0003-0.0004 ppm Ag and 0.004-0.005 ppm Bi for the nitrate samples. The coefficient of variation, in general, is 10-25% for Ag and 33% for Bi.     

Separation and preconcentration of trace amounts of several metals in magnesium metal and nitrate, with activated carbon as a collector

A method is described for the preconcentration and determination of traces of Hg, Ag, Cu, Fe, In, Mn, Pb, and Zn present as impurities in magnesium metal (1 g) and nitrate (100 g). After the metal sample has been dissolved in nitric acid (or the salt in water) and the pH adjusted to 8.1-9 (except for preconcentration of Hg, when pH 3 is used), the solution is filtered through a 2-cm paper coated with 50 mg of activated carbon. The trace metals are quantitatively adsorbed on the activated carbon and separated from the matrix. The rest of the procedure has already been described. The detection limits for the analysis of 1 g of Mg and 100 g of Mg(NO(3))(2).6H(2)O are 0.03-1.3 ppm and 0.00031-0.013 ppm respectively, for all the trace metals except Hg. The limit is 0.00001(4) ppm for Hg in 100 g of Mg(NO(3))(2).6H(2)O. The coefficient of variation is 4-33%, depending on the trace metal.     

Single and Sequential Extraction in Sediments and Soils

Abstract

The determination of extractable trace metal contents in soils and sediments is currently performed in many laboratories to assess the bioavailable metal fraction (and related potential phyto-toxic effects) and the importance and possibility of mobilization of trace metals from polluted soil, sludge and sediment upon landfill application. Single and sequential extraction schemes are used for the assessment of the different “forms” of trace metals (e.g. “mobile/bioavailable”, “carbonate-bound” etc.). The lack of uniformity in the different extraction procedures used throughout the world does not allow the results to be compared or the procedures to be validated which has led to many criticisms in the past few years. Moreover, the lack of suitable reference materials for this type of operationally defined determinations did not enable the quality of the measurements to be controlled. Owing to the need for establishing common schemes for single and sequential extractions as well as for the improvement of the quality of extractable trace metal determinations in soil and sediments, the Community Bureau of Reference (BCR) has organised a project which results along with the state of the art of extractable trace metal determinations, use and applicability of extraction schemes and analytical limitations were decided to be thoroughly discussed in a workshop. This paper presents its main conclusions.     

Determination of iron and nickel by flame atomic absorption spectrophotometry after preconcentration on Saccharomyces cerevisiae immobilized sepiolite

Iron and nickel have been preconcentrated on Saccharomyces cerevisiae immobilized sepiolite and determined by flame atomic absorption spectrophotometry (FAAS). Preconcentration studies were conducted by the column method. Effect of pH, amount of adsorbent, elution solution, flow rate and interfering ions on the recovery of the analytes have been investigated. Recoveries of Fe and Ni were 95+/-1 and 99.5+/-0.1%, respectively, at 95% confidence level. The breakthrough capacities of analytes were also investigated and found to be 0.042 mmol g(-1) for Fe and 0.055 mmol g(-1) for Ni. The proposed method was applied to the determination of iron and nickel in brass (NBS SRM 37e). The detection limit of iron and nickel were found as 0.065 and 0.087 mug ml(-1), respectively. The direct determination of trace metals by flame atomic absorption spectrometry (FAAS) is limited and difficult because of low concentration and/or matrix interferences. The proposed method is excellent for the determination of trace metal in matrixes, such as metal alloys.     

Determination of trace level impurities in uranium compounds by ICP-AES after organic extraction

The determination was studied of Al, B, Be, Cd, Ca, Co, Cu, Mg, Mn, Mo, Pb, Si, Sn, V, Cr, Ni, and Fe as trace level impurities in uranium compounds by ICP-AES after extraction of uranium with three different mixtures of di-(2-ethyl-hexyl) phosphate (D2EHP) and tri-(2-ethyl-hexyl)-phosphate (T2EHP) in solvents like toluene, carbon tetrachloride, hexane and cyclohexane. The study was carried out in presence of different concentrations of HCl and HNO(3). A single extraction with D2EHP in cyclohexane using nitric acid as matrix was sufficient to reduce the U(3)O(8) concentration from 100 g/l to 100 microg/ml. The ICP-AES instrumentation applied, allowed the determination of metal concentrations ten-times lower than those usually found in nuclear grade U(3)O(8). To check the efficiency of the extraction and the accuracy of the proposed method, Certified Reference Materials were used in the dissolution and extraction steps. The method described can be used for the determination of trace metals in nuclear grade U(3)O(8).     

Microwave assisted sample preparation for determining water-soluble fraction of trace elements in urban airborne particulate matter: evaluation of bioavailability

The feasibility of using two different microwave-based sample preparation methods was investigated to determine the total and water-soluble trace metal fraction in airborne particulate matter. The extraction techniques were then applied to urban particulate matter of different sizes in order to evaluate their bioavailability of associated trace metals. While a combination of HNO₃-HF-H₂O₂ was used for the total trace metal fraction of particulate matter, water was employed for the microwave-assisted extraction of water-soluble trace metal fractions. Inductively coupled plasma-mass spectrometry (ICP-MS) was used for the analysis of trace elements. The experimental protocol for the microwave assisted digestion was established using two different SRMs (1648, urban particulate matter and 1649a, urban dust). In the case of water-soluble trace metal fraction, the quantities extracted from the SRMs were compared between ultrasonic and microwave-assisted extractions, and there was a good agreement between the two extraction methods. Blanks values and limits of detection (LODs) for total and water-soluble trace metal concentrations were determined for three different filter substrates (Teflon, Zeflour, and Quartz). Subsequently, the proposed digestion method was evaluated for its extraction efficiency with these filter substrates. Finally, the real-world application of the proposed microwave-based sample preparation methods was demonstrated by analyzing trace elements in airborne particulate samples collected from different outdoor environments in Singapore. The solubility of 11 trace elements detected in the particulate samples is quantified.