氮掺杂碳量子点荧光法检测猪肉中的四环素

四环素（TC）作为一种广谱抗生素,用于预防和治疗细菌性疾病,以及作为饲料添加剂促进动物生长。本文旨在建立一种菠萝皮来源的氮掺杂碳量子点（N-CQDs）荧光猝灭法快速检测TC。以菠萝皮和二乙烯三胺为原料,通过水热法制备高量子产率的蓝色荧光N-CQDs。通过透射电子显微镜、傅里叶红外光谱、X射线光电子能谱对N-CQDs进行结构表征。结果表明,N-CQDs的平均粒径为3.53 nm,主要由C、N、O 3种元素组成,表面具有含氧和含氮基团。主要猝灭机理为静态猝灭和内滤效应。在最优试验条件下,该方法在0.3～40μg/mL范围线性良好,检出限为89.26μg/L。该检测方法被用于猪肉中四环素的检测,回收率在98.40%～106.02%之间,相对标准偏差1.29%～3.88%。     

胭脂红与牛血清白蛋白相互作用的光谱学研究

采用荧光光谱法与紫外光谱法研究胭脂红与牛血清白蛋白(BSA)之间的相互作用。荧光猝灭结果表明:在模拟生理条件(p H=7.4)下,胭脂红对蛋白质的动态猝灭常数为4.54×10~(13)L/(mol·s),属于静态猝灭;298K时其结合常数与结合位点数分别为5.33×10~7L/mol和1.35;热力学参数熵变(ΔH)与焓变(ΔS)均为负数,表明胭脂红与BSA之间的作用力为范德华力和氢键;依据Fster非辐射能量转移理论求得胭脂红与BSA之间的结合距离为3.75 nm,两者之间极有可能发生非辐射能量转移现象,导致荧光猝灭。同步荧光光谱法和三维荧光光谱法试验表明:胭脂红引起BSA分子构象的变化。     

食用合成色素胭脂红对胰蛋白酶光谱性质的影响

应用荧光光谱法、紫外可见吸收光谱法、傅立叶变换红外光谱和圆二色光谱法,研究了食用合成色素胭脂红与胰蛋白酶的结合性质,明确胭脂红与胰蛋白酶之间的结合常数、结合位点数（n）、作用力类型、结合距离（r）等信息。结果表明,胭脂红对胰蛋白酶内源荧光强度有较强的猝灭能力,荧光猝灭机制属于形成复合物的静态猝灭,由实验计算得出的热力学参数熵变和焓变均为负值,表明氢键和范德华力是驱动胭脂红-胰蛋白酶复合物形成的主要作用力,胭脂红与胰蛋白酶的结合常数达到104mol/L数量级,存在中等强度的亲和力。光谱分析表明胭脂红与胰蛋白酶结合引起胰蛋白酶的氨基酸残基微环境发生改变,胰蛋白酶的二级结构α-螺旋和无规则卷曲含量增加,β-折叠和β-转角的含量降低,多肽链有所收缩。      

荧光光谱法研究牛血清白蛋白与柠檬黄和日落黄的相互作用

用紫外-可见光吸收光谱、荧光发射光谱法研究牛血清白蛋白与柠檬黄和日落黄之间的相互作用;用Stern-Volmer方程处理实验数据,得到310 K时的动态猝灭速率常数（KQ）分别为2.51×105 L/mol和1.42×105 L/mol;用Lineweaver-Burk双倒数函数方程处理实验数据,得到310 K时静态猝灭常数（KLB）依次为5.79×105 L/mol和5.56×105 L/mol;用lg（（F0－F）/F）=lgK0+nlgρ处理实验数据,得到结合位点数（n）分别为0.556 95、0.640 56。并且两种色素在质量浓度为0～3 μg/mL范围内其含量与荧光猝灭强度具有良好的线性关系。     

亮橙色荧光碳点探针快速检测水产品中孔雀石绿

摘 要：目的 建立以亮橙色荧光碳点（bright orange fluorescent carbon dots, BO-CNDs）为探针的荧光检测法快速灵敏测定水产品中的孔雀石绿（malachite green,MG）。方法 对工作参数进行优化后,研究该检测方法的线性范围、灵敏度、选择性及抗干扰性,探究BO-CNDs和MG之间的相互作用,进而提出MG对BO-CNDs荧光猝灭的机制,最终将该方法应用到水产品中MG含量的测定。结果 在最佳检测条件下,BO-CNDs对MG检测的线性范围为0.01~10.00 μmol/L,检测限低至4.23 nmol/L,表现出反应快、选择性好、灵敏度高的优势。MG对BO-CNDs的荧光猝灭是内滤效应（inner filter effect, IFE）和动态猝灭协同作用的结果。最终将该检测方法应用于白鲢鱼、黑鱼、武昌鱼、鲤鱼、草鱼等鱼类样品中MG含量的测定,回收率在96.00%~102.00%之间,相对标准偏差(relative standard deviations, RSDs)低于1.01%。结论 该方法检测限低、灵敏度高、准确性好,具有应用于水产品中MG快速检测的巨大潜力。     

硫代乙醇酸修饰CdS量子点荧光猝灭法测定呋喃西林

为了对水产品中呋喃西林进行简单、灵敏分析,本论文建立了一种检测呋喃西林的荧光分析方法。以硫代乙醇酸修饰的CdS量子点(TGA-CdS QDs)为荧光探针,在pH7.0的磷酸盐缓冲溶液中,呋喃西林使TGA-CdS QDs荧光猝灭,据此建立了基于TGA-CdS QDs荧光猝灭法检测呋喃西林含量的新方法。在最佳实验条件下,检测线性范围为2.0×10-6~1.0×10-4mol/L,线性回归方程F0/F=17467 C(mol/L)+0.9918,相关系数为0.9984,检出限为7.12×10-7mol/L。方法应用于小虾呋喃西林含量的测定,回收率在95.6%~97.0%之间。     

色氨酸荧光猝灭法测定海带中碘离子

研究碘离子对色氨酸的荧光猝灭效应,旨在建立色氨酸荧光猝灭法测定海带中碘离子。在p H=6.0的磷酸缓冲液中,加入不同浓度的碘离子,测定色氨酸溶液的荧光光谱及强度的变化。试验表明,色氨酸具有释放荧光的特性,最佳激发和发射波长分别为223 nm和352 nm。碘离子对色氨酸的荧光具有猝灭作用。色氨酸浓度为10μmol/L时,碘离子对色氨酸的荧光猝灭呈线性关系,根据猝灭程度可计算出碘离子的浓度,该方法的检测限为1.64μmol/L。根据该方法检测了两种海带样品中的碘离子,其浓度为637μg/g干重(Laminaria japonica Aresch)和621μg/g干重(Saccharina japonica)。     

氮掺杂碳点的开关型荧光传感器检测蔬菜中的生物硫醇

研究并建立一种开关型荧光传感器模型快速检测Hg²⁺和生物硫醇。以柠檬酸和尿素为前驱体,通过一锅水热法制备了氮掺杂碳点(N-CDs),并对其进行了表面形貌、紫外和荧光光谱以及表面基团表征的检测,以检测体系pH、Hg²⁺浓度、孵育时间(荧光猝灭和恢复时间)为单因素,优化检测生物硫醇的最佳条件,并在最优检测条件下建立相应的标准曲线。结果表明,N-CDs的最佳激发和发射峰分别是340和432 nm,量子产率(FLQY)为32.72%。优化得到谷胱甘肽(GSH)的最佳检测条件为pH=5.0,Hg²⁺浓度为200 μmol/L,460 s荧光猝灭和380 s荧光恢复,最佳检测半胱氨酸(Cys)条件为pH=5.6,Hg²⁺浓度为300 μmol/L,440 s荧光猝灭和400 s荧光恢复,在最优条件下该传感器对0~300 μmol/L的GSH和100~400 μmol/L的Cys具有线性响应,检测限(LOD)分别为2.56和3.46 μmol/L,加标回收率为80%~120%。以上结果表明该传感器对检测蔬菜中生物硫醇有较好的选择性和准确度,能为食品基质中生物硫醇的荧光快速检测提供一种新思路。     

基于脱氧核酶-荧光共振能量转移效应的食品中铅离子快速检测

为实现食品中铅（Pb2+）污染的快速检测,该研究构建了一种基于Pb2+依赖型脱氧核酶（DNAzyme）及荧光共振能量转移（Fluorescence resonance energy transfer,FRET）效应的“Turn on”型铅离子检测传感器。荧光猝灭基团BHQ1可以作为FRET受体猝灭EvaGreen的荧光。当Pb2+存在时,底物链的特异性切割反应可以减弱BHQ1对EvaGreen的猝灭效果,BHQ1随底物链的断裂而释放,远离供体EvaGreen,因此FRET效应减弱,供体EvaGreen的荧光得以保留。该研究利用Pb2+诱导的FRET效应的减弱及EvaGreen荧光信号的增强可以对铅离子进行定性、定量分析。当脱氧核酶臂长为15 nt~5 nt、EvaGreen浓度为1×、底物链与酶链的浓度分别为400 nmol/L与100 nmol/L时,探究了该方法的检出限及线性范围。该反应可在室温条件下3 min内完成,同时对铅离子具有高度的特异性,可以区分不同的金属离子,并且成功应用于新鲜鸡蛋及自来水中的铅离子定量分析,加标回收率为86.79%~113.32%。该研究设计的“Turn on”型铅离子检测传感器可于室温条件下进行一管混反应,无需扩增,反应迅速,为检测食品中的铅污染提供了一些思路。     

基于DNA保护的银纳米簇荧光探针检测啶虫脒

目的设计合成DNA-AgNCs荧光探针用于检测啶虫脒。方法以啶虫脒的适配体DNA为保护剂,硼氢化钠为还原剂还原AgN03,合成发射波长为421nm的DNA-AgNCs。啶虫脒溶液与该纳米簇混合时,荧光强度降低,根据该现象检测啶虫脒。结果在37℃的条件下,啶虫脒于DNA-AgNCs体系中(在pH=7.0的磷酸缓冲溶液中)反应60 min后荧光猝灭,啶虫脒的浓度在5～200μmol/L范围内呈现出良好的线性关系,检出限为2.8μmol/L(S/N=3)。结论该检测方法较简单,易操作,快速、准确、灵敏,适用于测定啶虫脒。     

离子色谱法同时测定蛋白胨中葡萄糖、果糖、蔗糖、麦芽糖、乳糖含量

目的建立离子色谱法同时检测蛋白胨中葡萄糖、果糖、蔗糖、麦芽糖、乳糖的分析方法。方法样品用水作为提取剂,用OnGuard Ⅱ RP柱和OnGuard Ⅱ H柱净化,50mmol/LNaOH溶液作为流动相,用Carbopac PA20柱进行离子色谱分析。结果在该条件下5种糖都能得到较好分离,线性良好,葡萄糖、果糖、蔗糖、乳糖和麦芽糖的检出限为0.002、0.004、0.011、0.007、0.018 mg/L,平均回收率为90.59%～97.68%(n=9)。结论该方法操作简便、灵敏度高,可用于作为蛋白胨中葡萄糖、果糖、蔗糖、麦芽糖、乳糖的检测。     

拟穴青蟹新型过敏原肌质钙结合蛋白的纯化、鉴定及分子克隆

为进一步认识蟹类的过敏原,采用免疫印迹方法,分析发现甲壳类过敏患者血清能与拟穴青蟹肌浆蛋白中分子质量约为21 kD的蛋白质产生特异的IgE结合反应,结果显示该蛋白可能是蟹类新型过敏原。通过硫酸铵盐析、阴离子交换和凝胶过滤柱层析等方法对21 kD-蛋白进行分离纯化,采用Western blotting和基质辅助激光解析电离飞行时间质谱（matrix assisted laser desorption ionization-time of flight-mass spectrometry,MALDI-TOF-MS）确认纯化的21 kD-蛋白为肌质钙结合蛋白（sarcoplasmic calcium binding protein,SCP）。采用SMART-RACE（SwitchingMechanism At RNA Termini-Rapid Amplification of cDNA Ends）的方法获得SCP的cDNA序列,该序列全长986 bp,开放阅读框为579 bp,编码193 个氨基酸,其理论分子质量21.94 kD,等电点4.44。拟穴青蟹SCP与甲壳类动物SCP具有较高的同源性,与昆虫SCP的同源性较差;三级结构模拟分析显示,SCP含有5 个螺旋-转角-螺旋结构区,即EF-手型结构区,并在其中两个手型结构区形成2 个钙离子结合位点;进一步预测得到SCP的4 个线性抗原表位和3 个构象性抗原表位。     

Impact of transglutaminase on the textural, physicochemical, and structural properties of chicken skeletal, smooth, and cardiac muscles

This study examined the effects of microbial transglutaminase (MTG; 3.1 mg/ml) on chicken skeletal, smooth, and cardiac muscles; the meat containing the different muscle types was shaped into sausages and treated at 40 °C and/or 78 °C for 30 min. Although the three muscle types were obtained from the same bird, the effects of MTG addition were not uniform. All the muscle types showed a significant increase in the breaking strength (P < 0.01), but skeletal muscle exhibited the maximum increase. All samples showed a decrease in the fluorescence intensity and a significant reduction in the concentration of proteins that were extracted in a high ionic strength solution (P < 0.05). Scanning electron microscopy images and histological studies revealed that different muscle types had different physical structures and frameworks after MTG treatment, which is a reflection of the differences in the reaction specificity of MTG with different muscle proteins. Histological studies revealed that the reactions of MTG with meat proteins are both exogenous and endogenous. Cooking loss data suggested that MTG did not have any negative effect on water retention during cooking. MTG appears to be a functional and contributive substance since the results suggest that MTG can function on all muscle types that are mechanically processed for different industrial applications. MTG aggregates muscle proteins in different ways that improve their organoleptic properties such as texture, appearance, and water retention.     

Determination of Zn,Cu, Cd,Pb, Ni and Cr in some Slovenian foodstuffs

An acid-assisted microwave digestion procedure was optimised for the determination of trace elements in foodstuffs by the use of flame and electrothermal atomic absorption spectrometric (FAAS, ETAAS) techniques. Zn, Cu, Cd, Pb, Ni and Cr were determined in cabbage, wheat, potato, instant milk, fish, eggs and baby foodstuffs. The repeatability of measurements was tested in the cabbage sample and was found to be ±3.3% for Zn, 4.6% for Cd, ±6.8% for Cu and ±15.5% for Cr. The worse value obtained for Cr was probably due to its inhomogeneous distribution and very low concentration in the cabbage. Concentrations of Ni and Pb in the cabbage sample were below the limit of detection (0.2 mg kg^–1). The accuracy of the analytical procedure was checked by the analysis of the standard reference material CRM 1570 Trace and minor elements in spinach leaves and IAEA 336 Trace and minor elements in lichen. Good agreement between the certified or indicative and determined values was obtained. The results indicate that the concentrations of Zn, Cu, Cd, Pb, Ni and Cr found in cabbage, wheat and potato are comparable to the literature data. Cd, Pb and Ni were not found to be the contaminants in the foodstuffs analysed. Higher concentrations of Ni were found only in baby food "okolenik", arising from the cocoa that "okolenik" contained. It was demonstrated that the Cr content is very low in baby food, yolk and white of egg and in fish (<0.05 mg kg^–1 Cr). Elevated concentrations of trace elements were found only in mussels that accumulate heavy metals from the marine environment.     

Pilot survey of commercial pasteurized milk consumed in the metropolitan area of Rio de Janeiro,Brazil, for tetracyclines residues,including the 4-epimers of oxytetracycline,tetracycline and chlortetracycline

This pilot survey aimed to assess the occurrence of tetracyclines and the 4-epimers of oxytetracycline, tetracycline and chlortetracycline in commercial pasteurized milks sold in the metropolitan area of Rio de Janeiro, Brazil, between October 2009 and March 2010. A liquid chromatography–tandem mass spectrometry (LC–MS/MS) method, developed and validated in our laboratory, was used. All 100 analyzed samples were compliant, but 14 contained oxytetracycline in concentrations ranging from the method limit of detection (3.7?µg?l^–1) to the method limit of quantification (12.2?µg?l^–1). One sample contained oxytetracycline and tetracycline simultaneously (at a concentration slightly higher than method limit of quantification, 7.0?µg?l^–1). The presence of 4-epioxytetracycline and 4-epitetracycline in contaminated samples with the parent drugs could not be confirmed as traces were detected only in the quantification MRM transition. No other tetracyclines were detected.     

超声波对低嘌呤脱脂豆浆溶解性、保水性及粘度的影响

为了研究超声波处理对低嘌呤脱脂豆浆溶解性、保水性及黏度的影响,以脱脂豆粕为原料,采用超声波法,通过单因素及正交试验,研究最佳超声波处理条件.研究结果表明,最佳超声波处理条件为超声功率315 W、超声时间40 min、超声温度60℃,此条件下的黏度为1.4 mPa·s、保水性为81.83％、溶解性为58.15％.超声波处理的溶解性比未经超声波处理提高了22.35％.因此,超声波处理能显著提高低嘌呤脱脂豆浆的溶解性及低嘌呤脱脂豆腐的保水性,降低低嘌呤脱脂豆浆的黏度.     

Bioactive compound content,antioxidant activity,deoxynivalenol and heavy metal contamination of pearled wheat fractions

Wheat kernels are rich in antioxidant compounds, that are mainly present in the outer bran layers and which are removed during milling. Unfortunately, several contaminants, e.g., mycotoxins and heavy metals, are also concentrated in the external layers. Pearling of 3 wheat varieties gave five fractions (each 5% of the original grain weight), starting from the outer layer until the inner kernel, designated as 0–5%, 5–10%, 10–15%, 15–20%, 20–25%, respectively. The remaining 75% of the inner kernel was also collected. Dietary fibre, free phenolic acids and total antioxidant activity decreased progressively from the external to the internal layers. However, the 5–10% fraction was richer in β-glucan content than the external one (0–5%). Heavy metals were only found in the most external fraction. Deoxynivalenol contamination decreased from the external to the internal layers: 64% of total contamination of kernel was found in the 0–5% and 5–10% fractions. The 10–15% kernel fraction offered the best compromise between high nutritional value and low contamination risk.     

红发夫酵母中3R,3’R-虾青素的分离纯化和结构鉴定

研究红发夫酵母中3R,3’R-虾青素的分离纯化工艺条件,并利用高效液相色谱串联大气压化学电离源质谱(high performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry,HPLCAPCI-MS)、核磁共振(nuclear magnetic resonance,NMR)和高效液相色谱-紫外光谱(high performance liquid chromatography-ultra violet,HPLC-UV)对纯化得到的虾青素结构进行鉴定。结果表明:利用细胞玻璃研磨器进行研磨提取、硅胶柱层析分离和结晶提纯相结合的方法,能够有效减少反式虾青素的异构化,得到的虾青素纯度高于95%。采用高效液相色谱-质谱仪(high performance liquid chromatography-mass spectrometry,HPLC-MS)、超导核磁共振谱仪(1H和13C NMR)以及Chiralpak IC手性固定化纤维柱分析,显示制备的反式虾青素几乎都以3R,3’R结构组成。     

高效液相色谱法同时测定可乐中苯甲酸、糖精钠、安赛蜜和咖啡因

目的建立高效液相色谱同时测定可乐中苯甲酸、糖精钠、安赛蜜和咖啡因的方法,并应用于国际FAPAS能力验证。方法样品经超声脱气用水稀释,采用C_(18)色谱柱分离,以乙酸铵缓冲液(0.02 mol/L)/甲醇为流动相,采用梯度洗脱分离,在230 nm、272 nm波长下进行检测。结果苯甲酸、糖精钠、安赛蜜和咖啡因线性范围为2.0~200μg/m L,线性关系良好,相关系数大于0.999,回收率在97%~103%之间,RSD为0.5%~2.8%之间,检出限在0.005~0.02μg/m L之间。结论该法灵敏度高、重现性好,用于FAPAS能力验证时结果接近中值。