Objective: A high-throughput detection method for Cu2+ was developed with a well designed water-soluble fluorescent probe(HWEHH) and the microplate reader. Methods: The specificity of the fluorescent probe for Cu2+ determination was studied with the selectivity experiment. The linear range and detection limit of the probe for Cu2+ determination was detected with titration experiment, and the accuracy of the probe for the detection of Cu2+ in water was studied with sample spiked recovery method. Results: The fluorescent probe was able to specifically detect Cu2+. When the concentration of peptide probe was 2 μmol/L, the linear range of Cu2+ detection was 0~1.5 μmol/L. The linear equation was Y=1 135.085 7X+2 048.187 6, and the detection limit was 0.036 μmol/L. The binding constant of HWEHH-Cu2+ complex was 1.3×106 mol/L. The linear range of high-throughput detection method was 0~1.5 μmol/L, and the recovery rate was 93.6%~101.9%. Conclusion: HWEHH peptide fluorescent probe can be used for high-throughput detection of Cu2+ in aqueous solution.
Di-ya, XU; Hai-man, ZHANG; Xin, ZHANG; Ya-dan, ZHANG; Fang, WANG; and Lin, ZHANG
"High-throughput detection of copper ions in water by polypeptide probe HWEHH,"
Food and Machinery: Vol. 38:
8, Article 11.
Available at: https://www.ifoodmm.cn/journal/vol38/iss8/11
 鲍羽.微量元素与人体的健康[J].郧阳师范高等专科学校学报,2015,35(6):16-19.BAO Y.Trace elements and human health[J].Journal of Hanjiang Normal University,2015,35(6):16-19.
 BYUNG-EUN K,TRACY N,DENNIS J T.Mechanisms for copper acquisition,distribution and regulation[J].Nature Chemical Biology,2008,4(3):176-185.
 MOSTAFA M M,RAHMA E H,RADY A H.Chemical and nutritional changes in soybean during germination[J].Food Chemistry,1987,23(4):257-275.
 RODRIGUEZ M R,BALSA L M,PIRO O E,et al.Synthesis,crystal structure,spectroscopic characterization,DFT calculations and cytotoxicity assays of a new Cu(II)complex with an acylhydrazone ligand derived from thiophene[J].Inorganics,2021,9(2):10.
 贾斌,李漫,郑嘉,等.一种高效检测铜离子的罗丹明类荧光探针的构建及应用[J].化学试剂,2021,43(3):370-375.JIA B,LI M,ZHENG J,et al.Construction and application of a rhodamine fluorescent probe with high efficiency for detecting copper ions[J].Chemical Reagents,2021,43(3):370-375.
 LU Z W,DAI W L,LIU B C,et al.One pot synthesis of dandelion-like polyaniline coated gold nanoparticles composites for electrochemical sensing applications[J].Journal of Colloid and Interface Science,2018,525:86-96.
 赵国欣,赵明,李领川.环境水中重金属离子的现代检测方法研究综述[J].中州大学学报,2016,33(6):119-122.ZHAO G X,ZHAO M,LI L C.Review of modern detection methods of heavy metal ions in environmental water[J].Journal of Zhongzhou University,2016,33(6):119-122.
 辛思洁,林金石.土壤重金属检测方法应用现状及发展趋势[J].福建分析测试,2018,27(3):32-37.XIN S J,LIN J S.Application status and development trend of soil heavy metal detection methods[J].Fujian Analysis & Testing,2018,27(3):32-37.
 贺丽.浅析多种分析方法在水环境中重金属离子检测中的应用[J].产业与科技论坛,2019,18(6):74-75.HE L.Analysis on the application of various analytical methods in the detection of heavy metal ions in water environment[J].Industrial & Science Tribune,2019,18(6):74-75.
 袁敏,武建超,于劲松,等.水中重金属检测方法的研究进展[J].应用化工,2015,44(4):724-728.YUAN M,WU J C,YU J S,et al.Research progress of detection methods of heavy metals in water[J].Applied Chemical Industry,2015,44(4):724-728.
 王书源,李忠海,付湘晋,等.高荧光Cd Te量子点荧光探针测定Cu2+[J].食品与机械,2015,31(2):125-129.WANG S Y,LI Z H,FU X J,et al.Water soluable Cd Te quantum dots as fluorescence probes for copper(Ⅱ)detection[J].Food & Machinery,2015,31(2):125-129.
 马婧怡,田冰,王鑫,等.基于核酸适配体检测动物性食品中卡那霉素残留研究进展[J].食品与机械,2021,37(12):188-196.MA J Y,TIAN B,WANG X,et al.Research progress in aptasensors for the detection of kanamycin residues in animal-derived foods[J].Food & Machinery,2021,37(12):188-196.
 CUI J,ZANG S P,NIE H L,et al.An ICT-based fluorescent probe for ratiometric monitoring the fluctuations of peroxynitrite in mitochondria[J].Sensors and Actuators:B Chemical,2021,328:129069.
 LIANG L J,LIU C,JIAO X J,et al.A highly selective and sensitive photoinduced electron transfer(PET)based HOCl fluorescent probe in water and its endogenous imaging in living cells[J].Chemical Communications(Cambridge,England),2016,52(51):7 982-7 985.
 JIA X T,CHEN Q Q,YANG Y F,et al.FRET-based mito-specific fluorescent probe for ratiometric detection and imaging of endogenous peroxynitrite:Dyad of Cy3 and Cy5[J].Journal of the American Chemical Society,2016,138(34):10 778-10 781.
 LIU Y,NIE J,NIU J,et al.Ratiometric fluorescent probe with AIE property for monitoring endogenous hydrogen peroxide in macrophages and cancer cells[J].Scientific Reports,2017,7(1):7 293.
 SEDGWICK A C,WU L L,HAN H H,et al.Excited-state intramolecular proton-transfer(ESIPT)based fluorescence sensors and imaging agents[J].Chemical Society Reviews,2018,47(23):8 842.
 庞绪良.多肽基Cu2+,Hg2+荧光探针的合成和性能研究[D].聊城:聊城大学,2019:3-6.PANG X L.Synthesis and properties of polypeptide-based Cu2+ and Hg2+ fluorescent probes[D].Liaocheng:Liaocheng University,2019:3-6.
 戚平.食品和环境水中合成色素的高通量筛查及快速检测技术研究[D].广州:华南理工大学,2014:47-62.QI P.Research on high-throughput screening and rapid detection technology of synthetic pigments in food and environmental water[D].Guangzhou:South China University of Technology,2014:47-62.
 YURI O,KANAKO S,TAEKO T,et al.Fluorescent nucleic acid probe in droplets for bacterial sorting(FNAP-sort)as a high-throughput screening method for environmental bacteria with various growth rates[J].PLoS One,2019,14(4):533.
 黎卫强.紫外可见分光光度计在食品检测中的应用[J].企业科技与发展,2010(6):15-16.LI W Q.Application of UV-Vis spectrophotometer in food detection[J].Sci-Tech & Development of Enterprise,2010(6):15-16.
 MAIRI N C,JIANG D,WAIN A J,et al.Mechanistic studies of Cu(II)binding to amyloid-beta peptides and the fluorescence and redox behaviors of the resulting complexes[J].The Journal of Physical Chemistry B,2008,112(28):804611.
 ZHANG Y D,CAI Y H,HE Y H,et al.A label-free fluorescent peptide probe for sensitive and selective determination of copper and sulfide ions in aqueous systems[J].RSC Advances,2021,11(13):7 426-7 435.
 秦翠芳,李磊,俞宪同,等.含单个色氨酸的多肽分子的荧光特性研究:pH及金属离子的响应[J].光谱学与光谱分析,2017,37(2):476-480.QIN C F,LI L,YU X T,et al.Study on the fluorescence characteristics of peptide molecules containing single tryptophan:The response of pH and metal ions[J].Spectroscopy and Spectral Analysis,2017,37(2):476-480.
 张碧波.荧光小分子及荧光多肽探针在分析检测中的设计和应用[D].北京:清华大学,2017:1-5.ZHANG B B.Design and application of fluorescent small molecule and fluorescent polypeptide probe in analysis and detection[D].Beijing:Tsinghua University,2017:1-5.