Abstract
The hepatotoxic microcystins, especially microcystin–LR (MC–LR), are causing serious problems to public health and fisheries. We describe here a label-free amperometric immunosensor for rapid determination of MC–LR in water sample. The sensor was prepared by immobilizing antibody on a gold electrode coated with L-cysteine-modified gold nanoparticles. The stepwise self-assembly of the immunosensor was monitored and characterized by means of electrochemical impedance spectroscopy and differential pulse voltammetry. A 0.60 mmol L−1 solution of hydroquinone was used as the electron mediator. The immunosensor was incubated with MC–LR at 25 °C for 20 min, upon which the differential pulse voltammetric current changed linearly over the concentration range from 0.05 to 15.00 μg L−1, with a detection limit of 20 ng L−1. The developed biosensor was used to determine MC–LR in spiked crude algae samples. The recovery was in the range from 95.6 to 105%. This method is simple, economical and efficient, this making it potentially suitable for field analysis of MC-LR in crude algae and water samples.
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Falconer I, Bartram J, Chorus I, Kuiper-Goodman T, Utkilen H, Burch M, Codd GA (1999) In: Chorus I, Bartram J (eds) Toxic cyanobacteria in water. E & FN Spon, London, p 155
Gupta N, Pant SC, Vijayaraghavan R, Rao PVL (2003) Comparative toxicity evaluation of cyanobacterial cyclic peptide toxin microcystin variants (LR, RR, YR) in mice. Toxicology 188:285
World Health Organization (WHO) (1998) Guidelines for drinking water quality, 2nd edn. World Health Organization, Geneva
Meriluoto J (1997) Chromatography of microcystins. Anal Chim Acta 352:277
Kotak BG, Kenefick SL, Fritz DL, Rousseaux CG, Prepas EE, Hrudey SE (1993) Occurrence and toxicological evaluation of cyanobacterial toxins in Alberta lakes and farm dugouts. Water Res 27:495
Metcalf JS, Hyenstrand P, Beattie KA, Codd GA (2000) Effects of physicochemical variables and cyanobacterial extracts on the immunoassay of microcystin-LR by two ELISA kits. J Appl Microbiol 89:532
An J, Carmichael WW (1994) Use of a colorimetric protein phosphatase inhibition assay and enzyme linked immunosorbent assay for the study of microcystins and nodularins. Toxicon 32:1495
Mathysa W, Surholta B (2004) Analysis of microcystins in freshwater samples using high performance liquid chromatography and an enzyme–linked immunosorbent assay. J Hyg Environ Health 207:601
Tong P, Zhang L, He Y, Tang S, Cheng J, Chen G (2010) Analysis of microcystins by capillary zone electrophoresis coupling with electrospray ionization mass spectrometry. Talanta 82:1101
Zeisbergerova M, Kostal V, Sramkova M, Babica P, Blaha L, Glatz Z, Kahle V (2006) Separation of microcystins by capillary electrochromatography in monolithic columns. J Chromatogr B 841:140
Frias HV, Mendes MA, Cardozo KHM, Carvalho VM, Tomazela D, Colepicolo P, Pinto E (2006) Use of electrospray tandem mass spectrometry for identification of microcystins during a cyanobacterial bloom event. Biochem Biophys Res Commun 344:741
Tang D, Xia B (2008) Electrochemical immunosensor and biochemical analysis for carcinoembryonic antigen in clinical diagnosis. Microchim Acta 163:41
Zhang L, Liu Y, Chen T (2009) Label-free amperometric immunosensor based on antibody immobilized on a positively charged gold nanoparticle/L-cysteine-modified gold electrode. Microchim Acta 164:161
Lv P, Min L, Yuan R, Chai Y, Chen S (2010) A novel immunosensor for carcinoembryonic antigen based on poly (diallyldimethylammonium chloride) protected prussian blue nanoparticles and double-layer nanometer-sized gold particles. Microchim Acta 171:297
Tang D, Tang J, Su B, Chen H, Huang J, Chen G (2010) Highly sensitive electrochemical immunoassay for human IgG using double-encoded magnetic redox-active nanoparticles. Microchim Acta 171:457
Campas M, Marty JL (2007) Highly sensitive amperometric immunosensors for microcystin detection in algae. Biosens Bioelectron 22:1034
Zhang FH, Yang SH, Kang TY, Cha GS, Nam H, Meyerhoff ME (2007) A rapid competitive binding nonseparation electrochemical enzyme immunoassay (NEEIA) test strip for microcystin-LR (MC-LR) determination. Biosens Bioelectron 22:1419
Zhang J, Lei J, Xu C, Ding L, Ju H (2010) Carbon nanohorn sensitized electrochemical immunosensor for rapid detection of microcystin-LR. Anal Chem 82:1117
Wang F, Hu S (2009) Electrochemical sensors based on metal and semiconductor nanoparticles. Microchim Acta 165:1
Wu S, Zhong Z, Wang D, Li M, Qing Y, Dai N, Li Z (2009) Gold nanoparticle-labeled detection antibodies for use in an enhanced electrochemical immunoassay of hepatitis B surface antigen in human serum. Microchim Acta 166:269
Mena ML, Yanez-Sedeno P, Pingarron JM (2005) A comparison of different strategies for the construction of amperometric enzyme biosensors using gold nanoparticle-modified electrodes. Anal Biochem 336:20
Wu Y, Hu S (2006) Voltammetric investigation of cytochrome c on gold coated with a self–assembled glutathione monolayer. Bioelectrochemistry 68:105
Macdonald JR (1987) Impedance spectroscopy. Wiley, New York
Stricks W, Kolthoff IM (1951) Polarographic investigations of reactions in aqueous solutions containing copper and cysteine (Cystine). I. Cuprous copper and cysteine in ammoniacal medium. The dissociation constant of cuprous cysteinate. J Am Chem Soc 73:1723
Tudos AJ, Johnson DC (1995) Dissolution of gold electrodes in alkaline media containing cysteine. Anal Chem 67:557
Yang WR, Gooding JJ, Hibbert DB (2001) Characterisation of gold electrodes modified with self-assembled monolayers of L–cysteine for the adsorptive stripping analysis of copper. J Electroanal Chem 516:10
Long F, He M, Zhu A, Shi H (2009) Portable optical immunosensor for highly sensitive detection of microcystin-LR in water samples. Biosens Bioelectron 24:2346
Long F, Shi H, He M, Sheng J, Wang J (2009) Sensitive and rapid chemiluminescence enzyme immunoassay for microcystin-LR in water samples. Anal Chim Acta 649:123
Xia Y, Deng J, Jiang L (2010) Simple and highly sensitive detection of hepatotoxin microcystin-LR via colorimetric variation based on polydiacetylene vesicles. Sens Actuators B-Chem 145:713
Loyprasert S, Thavarungkul P, Asawatreratanakul P, Wongkittisuksa B, Limsakul C, Kanatharana P (2008) Label-free capacitive immunosensor for microcystin-LR using self-assembled thiourea monolayer incorporated with Ag nanoparticles on gold electrode. Biosens Bioelectron 24:78
Acknowledgements
The authors are grateful for the National Nature Sciences Foundation of China (20735002, 20877019, 21075016, 40940026), National Basic Research Program of China (2010CB732403), the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China (708056), the Nature Sciences Funding of Fujian Province (2009J1028, 2008J0228), China.
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Tong, P., Tang, S., He, Y. et al. Label-free immunosensing of microcystin-LR using a gold electrode modified with gold nanoparticles. Microchim Acta 173, 299–305 (2011). https://doi.org/10.1007/s00604-011-0557-8
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DOI: https://doi.org/10.1007/s00604-011-0557-8