Determination Of microcystin-LR Research Articles

Sensitive Identification of Microcystin-LR via a Reagent-Free and Reusable Electrochemical Biosensor Using a Methylene Blue-Labeled Aptamer

We report a methylene blue (MB)-modified electrochemical aptamer (E-AB) sensor for determining microcystin-LR (MC-LR). The signal transduction of the sensor was based on changes in conformation and position of MB induced by the binding between MC-LR and the modified aptamer probe. In the absence of MC-LR, an aptamer probe was considered partially folded. After combining aptamer and MC-LR, the configuration of the aptamer probe changed and facilitated the electron transfer between MB and the electrode surface. As a result, an increased current response was observed. We optimized the parameters and evaluated the electrochemical performance of the sensor using square wave voltammetry (SWV). MC-LR was measured from 1.0 to 750.0 ng/L with a detection limit of 0.53 ng/L. The reliability of the method was verified by the determination of MC-LR in environmental real samples, such as pond water and tap water. Moreover, we demonstrated that this reagent-less biosensor could be regenerated and reused after rinsing with deionized water with good accuracy and reproducibility. As a reusable and regenerable E-AB sensor, this rapid, reagent-free, and sensitive sensing platform will facilitate routine monitoring of MC-LR in actual samples.

Competitive Smartphone-Based Portable Electrochemical Aptasensor System Based on an Mxene/Cdna-Mb Probe for the Determination of Microcystin-Lr

Competitive Smartphone-Based Portable Electrochemical Aptasensor System Based on an Mxene/Cdna-Mb Probe for the Determination of Microcystin-Lr

Development of an impedimetric immunosensor to determine microcystin-LR. New approaches in the use of the electrochemical impedance spectroscopy was used in determining to determine kinetic parameters of immunoreactions

The development of new electrochemical immunosensors for the detection of environmental contaminants is of great interest due to their simplicity, high sensitivity, and extended analytical range. Because of the antibody is immobilized on the electrode, it is important to determine its loss of reactivity after immobilization. In this work, two aspects were addressed. First, we developed a new methodology based on electrochemical impedance (EI) to determine the kinetic parameters associated with immunoreactions carried out on the electrode. Second, an electrochemical immunosensor based on electrochemical impedance spectroscopy (EIE) was developed to determine microcystin-LR in drinking water samples. Microcystin-LR determination was based on a label-free non-competitive immunoassay. The electrochemical immunosensor shows a limit of detection (LOD) of 33 pg mL−1 (3.32 × 10−11 mol L−1 or 0.033 μg L−1) which is well below the WHO guideline recommendation of 1 μg L−1 and 40 times better than the LOD obtained using the same antibody in an optimized conventional competitive ELISA assay. In addition, an acceptable accuracy, with recovery percentages close to 100% were found. The label-free immunosensor is a valuable tool to monitor microcystin-LR in drinking water samples and the environment.

Ultrasensitive detection of microcystin-LR with gold immunochromatographic assay assisted by a molecular imprinting technique

A simple and highly sensitive lateral flow immunochromatographic assay (LFICA) towards microcystin-LR (MC-LR) was proposed in this study. Molecular imprinting technique was combined with enzyme assisted colorimetric method to substantially enhance the sensitivity of traditional LFICA. The target, i.e., MC-LR molecular, was first separated from the complex matrix by molecularly-imprinted polymers (MIPs). Then, the obtained MC-LR was detected by LFICA based on the blue color generated by an enzyme–substrate reaction. Using the proposed assay, under optimal conditions, the determination of MC-LR was doable in a wide linear range from 0.1 ng/mL to 100 ng/mL with a detection limit of 0.04 ng/mL. In addition, the reliability of the developed method was validated by determining MC-LR contents in real samples. The overall results suggested that the neoteric LFICA method possess great potential in sensitive detection of MC-LR.

Magnetic activated carbon@ iron oxide@manganese oxide composite as an adsorbent for preconcentration of microcystin –LR in surface water, tap water, water and wastewater

A simple, rapid and effective method based on dispersive solid phase microextraction (DSPME) and UV–vis spectrophotometer was developed for the preconcentration and determination of microcystin-LR (MC-LR) in environmental matrices. The activated carbon@ iron oxide@manganese oxide composite was used as solid phase material. The composite was characterized by scanning electron microscope/energy dispersive x-ray spectroscopy (SEM/EDS), transmission electron microscope (TEM), X-ray powder diffraction (XRD), nitrogen adsorption/desorption and Fourier transform infrared (FTIR). The factors affecting the DSPME were optimized using multivariate strategy. Under optimized condition, the LOD, LOQ, repeatability (n = 10), reproducibility (n = 5) and preconcentration factor were 0.5 μg L−1, 1.7 μg L−1, 3.1, 4.5% and 50, respectively. The percentage recovery of MC-LR in spiked water samples was found to be 98.5%. The developed method DSPME/UV–vis method was simple, rapid, cost-effective and can be used for the detection of MC-LR in real samples without any expensive or sophisticated equipment.

A pM leveled photoelectrochemical sensor for microcystin-LR based on surface molecularly imprinted TiO2@CNTs nanostructure

A simple and highly sensitive photoelectrochemical (PEC) sensor towards Microcystin-LR (MC-LR), a kind of typical cyanobacterial toxin in water samples, was developed on a surface molecular imprinted TiO2 coated multiwalled carbon nanotubes (MI-TiO2@CNTs) hybrid nanostructure. It was synthesized using a feasible two-step sol-gel method combining with in situ surface molecular imprinting technique (MIT). With a controllable core-shell tube casing structure, the resultant MI-TiO2@CNTs are enhanced greatly in visible-light driven response capacity. In comparison with the traditional TiO2 (P25) and non-imprinted (NI-)TiO2@CNTs, the MI-TiO2@CNTs based PEC sensor showed a much higher photoelectric oxidation capacity towards MC-LR. Using this sensor, the determination of MC-LR was doable in a wide linear range from 1.0pM to 3.0nM with a high photocurrent response sensitivity. An outstanding selectivity towards MC-LR was further achieved with this sensor, proven by simultaneously monitoring 100-fold potential co-existing interferences. The superiority of the obtained MC-LR sensor in sensitivity and selectivity is mainly attributed to the high specific surface area and excellent photoelectric activity of TiO2@CNTs heterojunction structure, as well as the abundant active recognition sites on its functionalized molecular imprinting surface. A promising PEC analysis platform with high sensitivity and selectivity for MC-LR has thus been provided.

Upconversion nanoparticles grafted molybdenum disulfide nanosheets platform for microcystin-LR sensing

Water safety is one of the most pervasive problems afflicting people throughout the world. Microcystin-LR (MC-LR), a representative toxin released by cyanobacteria, poses an increasing and serious threat to water safety. In order to develop facile, specific and sensitive detection methods for MC-LR, we fabricated an ultrasensitive fluorescence aptasensor using the enhanced fluorescence of UCNP and the effective quenching ability, high affinity toward single strand DNA (ssDNA) of MoS2 (termed as FAUM). This assay specifically determined MC-LR in the linear range of 0.01–50ng/ml with a limit of detection (LOD) of 0.002ng/ml. The real water sample results indicated that this FAUM assay owns well enough reliability and feasibility to allow the determination of MC-LR. This aptamer-based method might be a promising strategy for a variety of sensing applications.

Determination of microcystin-LR in clams (Tapes decussatus) of two Sardinian coastal ponds (Italy)

The presence of microcystin-LR (MC-LR) was monitored in Tapes decussatus harvested in two Sardinian ponds (Cabras and Tortolì, Italy) in spring and summer. After solid phase extraction, samples were analyzed using a screening enzyme-linked immunosorbent assay (ELISA) followed by a liquid chromatographic coupled to tandem mass spectrometer (LC–MS/MS) analysis. Results obtained through the ELISA test showed the presence of microcystins with a maximum concentration in August for Cabras pond (0.55ng/g) and in September for Tortolì pond (0.85ng/g). The LC–MS/MS analysis did not confirm the presence of MC-LR suggesting that results obtained with the ELISA technique could be due to the presence of other microcystins. According to the tolerable daily intake suggested by the World Health Organization, these results hint that clams harvested in these ponds are safe for human health. These data can contribute to enrich the knowledge about the healthiness of Sardinian ponds and of their products.

Determination of microcystin-LR in surface water by a magnetic bead-based colorimetric immunoassay using antibody-conjugated gold nanoparticles

Herein we describe the development of a homogeneous competitive colorimetric immunoassay using antigen-functionalized magnetic beads (MBs) and antibody-immobilized gold nanoparticles (AuNPs) combined with the established gold staining method for the determination of microcystin-leucine-arginine (MC-LR) in surface water.

Determination of Microcystin-LR in Environmental Water by Magnetic Solid Phase Extraction-High Performance Liquid Chromatography

Abstract A rapid analytical method was established for the determination of trace amounts of microcystin (MC-LR) in environmental water samples by using magnetic solid phase extraction (MSPE) coupled with HPLC/UV based on a magnetic bentonite sorbent fabricated by solvothermal synthesis method. Several factors influencing the extraction efficiency of MC-LR, such as sorbent amount, pH condition, sample volume, adsorption and desorption time, and desorption solvent, were investigated and optimized. Under the optimal extraction conditions, the limits of detection and quantification for the method were 0.75 and 1.5 ng L −1 , respectively, which was far below the guideline recommended by the World Health Organisation. The method showed a good linearity ( R 2 = 0.9993) in the concentration ranges of 0.25–250 μg L −1 for MC-LR. The MSPE sorbent was simple in preparation, low cost and magnetizable, and the developed MSPE-HPLC method was suitable for the rapid and sensitive determination of MC-LR in nature water samples.

Measurement of microcystin -LR in water samples using improved HPLC method.

Microcystins are a group of toxic compounds produced by freshwater cyanobacteria and cause diseases. World Health Organization has recommended a concentration of 1 µg/l for Microcystin-LR (MC-LR) in potable water as guideline value. The high performance liquid chromatography (HPLC) followed by C18 analytical column and ultra violet detector for detection of MC-LR. In this regard, 5 different concentrations of MC-LR solutions were injected into HPLC. MC-LR was detected in 5.33 minute retention time and Calibration curve was achieved with R2=0.988. Detection limit for this method was obtained by using acetonitrile solutions (32% and 55%) as a gradient run and a high silanol activity column equal to 0.02 µg/mL. Despite no acidic organic modifier being used in the mixture of solvents, the sensitivity of this method was appropriate for detection of MC-LR. Because of short retention time, reduction in number of solvents and high resolution and suitable sensitivity, this method is affordable and is fast for detection and determination of MC-LR in potable water.

Ultrasensitive immunoassay of microcystins-LR using G-quadruplex DNAzyme as an electrocatalyst

An electrochemical immunoassay for microcystin-LR (MC-LR) detection was developed using multi-labeled horseradish peroxidase-mimicking DNAzyme on carbon nanotubes (CNTs) as electrocatalyst for signal amplification. CNTs were covalently conjugated to multiple DNAzyme along with MC-LR for a competitive immunoassay. The as-prepared DNAzyme/CNTs/MC-LR biolabel was specifically captured on the electrode surface, and current responses were obtained upon the electro-catalytic reduction of hydrogen peroxide by the captured biolabels. Under optimal conditions, the electro-catalytic current decreased linearly with the increase amount of MC-LR in the range from 0.01 to 7.0 µg L−1. The linear regression equation was I (µA) = 12.96 − 1.48 X [MC–LR] (µg L−1), with a correlation coefficient of 0.989. The limit of detection of MC-LR was 2.31 ng L−1. Application of the immunoassay method and LC/MS/MS method for MC-LR determination on spiked reservoir water gave recovery range of 91.7–105.2% and 94.0–105.0%, respectively. The resulting versatile immunoassay exhibited high sensitivity, good precision and satisfactory reproducibility, which could have vast potential in routine water quality monitoring for various environmental toxins.

Determination of microcystin-LR in water by a label-free aptamer based electrochemical impedance biosensor

In this study, an electrochemical impedance biosensor for cyanobacterial toxin microcystin-LR (MC-LR) detection has been developed. MC-LR aptamers were immobilized on the gold electrode through Au–S interaction, in the presence of target (MC-LR); the binding of MC-LR and aptamers probe led to a complex formation change on the electrode surface and resulted in the impedance decreasing. The decrease rate had a linear relationship with logarithm of the MC-LR concentration in the range of 1.0×10−7–5.0×10−11mol/L, with a detection limit of 1.8×10−11mol/L. The sensor has good selectivity and stability, it has been applied to detect MC-LR in three kinds of real water samples with satisfying results.

Fabrication of a Novel and Simple Microcystin-LR Photoelectrochemical Sensor with High Sensitivity and Selectivity

Microcystin-LR (MC-LR), an inert electrochemical species, is difficult to be detected by a simple and direct electrochemical method. In the present work, a novel photoelectrochemical sensor is developed on highly ordered and vertically aligned TiO(2) nanotubes (TiO(2) NTs) with convenient surface modification of molecularly imprinted polymer (MIP) (denoted as MIP@TiO(2) NTs) for highly sensitive and selective determination of MC-LR in solutions. Molecularly imprinted polypyrrole (PPy) of MC-LR is chosen as the recognition element. The designed MIP@TiO(2) NTs photoelectrochemical sensor presents excellent applicability in MC-LR determination, with linear range from 0.5 to 100 μg L(-1) and limit of detection of 0.1 μg L(-1). Moreover, the sensor exhibits outstanding selectivity while used in coexisting systems containing 2,4-dichorophenoxyacetic acid, atrazine, paraquat, or monosultap with high concentration, 100 times that of MC-LR. The sensor presents good photoelectric conversion efficiency and detection sensitivity, as well as broad linear detection range, mainly because of the high specific surface area and photoelectric activity of TiO(2) NTs and the π bond delocalized electron system of PPy that promotes the separation of electron-holes. The prominent selectivity is from the MIP by forming multiple hydrogen bonds between PPy and MC-LR. Mechanisms for photoelectrochemical analysis and selective recognition are also discussed.

Selective Determination of Microcystin-LR Based on Proteasomal Trypsin-Like Activity Inhibition

A sensitive and selective method was developed for the determination of microcystin-LR (MC-LR) based on proteasomal trypsin-like activity inhibition. Under the optimized conditions, the detection limit for MC-LR was 0.18μg/L, The linearity range was 0.2-2 μg/L，and the average recovery for MC-LR in tap water was 128%. Microcystin-YR (MC-YR) and Microcystin-RR (MC-RR) have no inhibiting effect on the proteasomal trypsin-like activity in the concentration ranged between 0.1-100 nmol/L. This method could be applied for the selective, high-throughput, and rapid determination of MC-LR in waters.

Determination of Microcystin-LR in Drinking Water Using UPLC Tandem Mass Spectrometry-Matrix Effects and Measurement

A simple detection method using ultra-performance liquid chromatography electrospray ionisation tandem mass spectrometry (UPLC-ESI-MS-MS) coupled with the sample dilution method for determining trace microcystin-LR (MC-LR) in drinking water is presented. The limit of detection (LOD) was 0.04 µg/L and the limit of quantitation (LOQ) was 0.1 µg/L. Water matrix effects of ionic strength, dissolved organic carbon (DOC) and pH were examined. The results indicate that signal detection intensity for MC-LR was significantly suppressed as the ionic strength increased from ultrapure water condition, whereas it increased slightly with solution pH and DOC at low concentrations. However, addition of methanol (MeOH) into the sample was able to counter the signal suppression effects. In this study, dilution of the tap water sample by adding 4% MeOH (v/v) was observed to be adequate to compensate for the signal suppression. The recoveries of the samples fortified with MC-LR (0.2, 1, and 10 µg/L) for three different tap water samples ranged from 84.4% to 112.9%.

Determination of microcystin-LR and its metabolites in snail ( Bellamya aeruginosa), shrimp ( Macrobrachium nipponensis) and silver carp ( Hypophthalmichthys molitrix) from Lake Taihu, China

This paper describes seasonal changes of microcystin-LR (MC-LR) and its glutathione (MC-LR-GSH) and cysteine conjugates (MC-LR-Cys) in three aquatic animals – snail ( Bellamya aeruginosa), shrimp ( Macrobrachium nipponensis) and silver carp ( Hypophthalmichthys molitrix) collected from Lake Taihu, China. MC-LR, MC-LR-GSH, and MC-LR-Cys were determined by liquid chromatography electrospray ionization mass spectrum (LC–ESI-MS). The mean MC-LR concentrations in the hepatopancreas of snail and shrimp and liver of silver carp were 6.61, 0.24, and 0.027 μg g −1 dry weight (DW), respectively; while the average MC-LR-Cys concentrations were 0.50, 0.97, and 5.72 μg g −1 DW, respectively. MC-LR-GSH was usually not detectable in these samples. The above results suggest that: (1) in aquatic animals, especially fish, the main excretion form of MC-LR could be MC-LR-Cys, but not MC-LR-GSH, whereas MC-LR-Cys might play an important role in detoxication of MC-LR and (2) that efficiency of MC-LR-Cys formation differs among species. The main detoxication pathway of MC-LR in aquatic animals is suggested as follows: when MC-LR enters into liver/hepatopancreas, it firstly conjugates with polypeptide or protein (including GSH, PP-1 and 2A) containing Cys residues, perhaps also some free cysteine; subsequently, MC-LR-Cys is degraded from these polypeptide or protein; and finally is excreted from animals by the compound of MC-LR-Cys.

Determination of Microcystin-LR in Water from Lake Tai, China

Microcystin-LR (MC-LR) is a heptapeptide hepatotoxin produced by cyanobacteria. Immunoaffinity chromatography (IAC) column was prepared with CNBr-activated Sepharose 4B and monoclonal antibody of MC-LR. Water sample was cleaned up by IAC column and the content of MC-LR in water was determined by liquid chromatography-mass spectrometry (LC-MS). The results suggested that the IAC column exhibited highly selective specificity for MC-LR and selective removed interference from complex water sample. Water sample was concentrated for 2,000-fold through once purification. Cyanobacterial blooms had broken out in 2007 in Lake Tai, the third largest freshwater lake in China. Water samples from two parts of Lake Tai had been analyzed. The highest concentration of MC-LR in water from Lake Tai was 0.522 microg/L.

Monolithic solid-phase extraction for the rapid on-line monitoring of microcystins in surface waters

We describe the development and optimization of a sensitive and selective screening method for the measurement of trace levels of microcystins in surface waters. Several sample preparation techniques were compared, including solid-phase microextraction (SPME), particle-based solid-phase extraction (SPE), and monolith-based SPE. A flow-injection (FI) based approach employing a reversed-phase monolithic SPE column was found to be optimal. Quantification was performed by directly interfacing the FI-based SPE system to an electrospray ionization-mass spectrometer (ESI-MS). To more safely simulate peptidyl toxins such as the microcystins, a model peptide (i.e., angiotensin II) was used for method optimization. Sample loading flow rate and volume, eluent composition, and elution flow rate were optimized. Sample throughput was six samples per hour, a detection limit of 1.31 ng angiotensin II was demonstrated for a linear dynamic range from 1–1,000 ng and 3.4% relative standard deviation (n = 4, 100 ng sample). Sample volumes up to 1,000 ml of surface water could be loaded onto the monolithic SPE disk without exceeding the sorbent’s capacity. Unlike conventional particle-based SPE methods, the monolithic SPE disk does not need to be replaced between samples and could be used indefinitely. The FI-based SPE-ESI-MS method was successfully applied to the determination of microcystin-LR, the most common of the microcystins, in environmental samples and was demonstrated for the direct monitoring of chlorinated drinking water, with trends tracked over a period of eight months.

Determination of microcystin-LR in surface water using high-performance liquid chromatography/tandem electrospray ionization mass detector

The need for a rapid, sensitive, and reliable analytical method for microcystin-LR has been emphasized by the awareness of toxic cyanobacteria as a human-health risk through drinking water. Microcystin-LR is the most commonly reported microcystin which is produced by cyanobacteria. The WHO guideline for microcystin-LR in drinking water is 1μg/l. In this paper, an effective method has been developed by application of high-performance liquid chromatography/tandem electrospray ionization mass detector in the determination of microcystin-LR in surface water sample. At the LC–MS–MS CID–full scan mode, different relative collision energies have been tested with 30% being used for further microcystin-LR analysis. The possible mass dissociation path has been proposed. Based on 30% relative collision energy, present method has an excellent method detection limit (MDL), which is as low as 2.6ng/l. To the best of our knowledge, this represents one of the most sensitive methods in existence for the microcystin-LR analysis. This method has also been validated by evaluation of the calibration linearity, precision, accuracy, recovery, and mass ratio stability.