Comparison of Liquid Chromatography/Mass Spectrometry, ELISA, and Phosphatase Assay for the Determination of Microcystins in Blue-Green Algae Products

Lawrence, James F.; Niedzwiadek, Barbara; Ménard, Cathie; Lau, Benjamin P.‐Y.; Lewis, David A.; Kuper-Goodman, Tine; Carbone, Susan; Holmes, Charles F.B.

doi:10.1093/jaoac/84.4.1035

Cited by 83 publications

(50 citation statements)

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“…WHO alert level 2). Sample preparation involved a preliminary cyanobacteria cell lysis step with methanol (Lawrence et al, 2001) Frequency distribution analyses were carried out on grouped data with classes of 1, 2 and 5.10 i with i = -1; 0; 1; 2 for MCs concentrations, i= 4; 5; 6 for cell densities, and i= 5; 6; 7 for cell biovolumes.…”

Section: -Materials and Methodsmentioning

confidence: 99%

Improving microcystin monitoring relevance in recreative waters: A regional case-study (Brittany, Western France, Europe)

Pitois

Vezie²,

Thoraval³

et al. 2016

International Journal of Hygiene and Environmental Health

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Cyanobacteria and their toxins are known as a health hazard in recreative and distributed waters. Monitoring data from 2004 to 2011 were collected at regional scale to characterize exposition parameters to microcystins in Brittany (Western France). The data show that cyanobacteria populations are experiencing a composition shift leading to a longer duration of cell densities higher than WHO alert levels 2 and 3. Microcystins however appear to be more frequently detected with subacute concentrations in low cell density samples than in high cell density samples or during bloom episodes. Positive relations are described between microcystin concentrations, detection frequencies and cyanobacteria biovolumes, allowing for a novel definition of alert levels and decision framework following WHO recommendations.

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Section: -Materials and Methodsmentioning

confidence: 99%

Improving microcystin monitoring relevance in recreative waters: A regional case-study (Brittany, Western France, Europe)

Pitois

Vezie²,

Thoraval³

et al. 2016

International Journal of Hygiene and Environmental Health

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“…Different advances in LC-MS have resulted in lowered detection limits, such as the application of microbore LC coupled to an IT MS [27] for the analysis of microcystins from environmental samples. And not in the last place, the use of LC-MS/MS (multiple reaction monitoring, MRM) as a tool for toxin identification and quantification for regulatory purposes [28]. LC-MS/MS is relatively expensive for routine screening, and therefore initial analyses of the samples are often performed by ELISA.…”

Section: Determination/detectionmentioning

confidence: 99%

Toxins of cyanobacteria

Apeldoorn

Egmond²,

Speijers

et al. 2006

Molecular Nutrition Food Res

653

411

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Blue-green algae are found in lakes, ponds, rivers and brackish waters throughout the world. In case of excessive growth such as bloom formation, these bacteria can produce inherent toxins in quantities causing toxicity in mammals, including humans. These cyanotoxins include cyclic peptides and alkaloids. Among the cyclic peptides are the microcystins and the nodularins. The alkaloids include anatoxin-a, anatoxin-a(S), cylindrospermopsin, saxitoxins (STXs), aplysiatoxins and lyngbyatoxin. Both biological and chemical methods are used to determine cyanotoxins. Bioassays and biochemical assays are nonspecific, so they can only be used as screening methods. HPLC has some good prospects. For the subsequent detection of these toxins different detectors may be used, ranging from simple UV-spectrometry via fluorescence detection to various types of MS. The main problem in the determination of cyanobacterial toxins is the lack of reference materials of all relevant toxins. In general, toxicity data on cyanotoxins are rather scarce. A majority of toxicity data are known to be of microcystin-LR. For nodularins, data from a few animal studies are available. For the alkaloids, limited toxicity data exist for anatoxin-a, cylindrospermopsin and STX. Risk assessment for acute exposure could be relevant for some types of exposure. Nevertheless, no acute reference doses have formally been derived thus far. For STX(s), many countries have established tolerance levels in bivalves, but these limits were set in view of STX(s) as biotoxins, accumulating in marine shellfish. Official regulations for other cyanotoxins have not been established, although some (provisional) guideline values have been derived for microcystins in drinking water by WHO and several countries.

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“…1999;Gilroy et al. 2000;Lawrence et at, 2001 ). MCs are the most commonly found group of cyclic heptapeptide cyanotoxins with molecular weights ranging between 900 and 1100 Dalton (Da).…”

Section: Introductionmentioning

confidence: 99%

Oatp-associated uptake and toxicity of microcystins in primary murine whole brain cells

Feurstein

Holst

Fischer

et al. 2009

Toxicology and Applied Pharmacology

134

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Microcystins (MCs) are naturally occurring cyclic heptapeptides that exhibit hepato-, nephro- and possibly neurotoxic effects in mammals. Organic anion transporting polypeptides (rodent Oatp/human OATP) appear to be specifically required for active uptake of MCs into hepatocytes and kidney epithelial cells. Based on symptoms of neurotoxicity in MC-intoxicated patients and the presence of Oatp/OATP at the blood-brain-barrier (BBB) and blood-cerebrospinal-fluid-barrier (BCFB) it is hypothesized that MCs can be transported across the BBB/BCFB in an Oatp/OATP-dependent manner and can induce toxicity in brain cells via inhibition of protein phosphatase (PP). To test these hypotheses, the presence of murine Oatp (mOatp) in primary murine whole brain cells (mWBC) was investigated at the mRNA and protein level. MC transport was tested by exposing mWBCs to three different MC-congeners (MC-LR, -LW, -LF) with/without co-incubation with the OATP/Oatp-substrates taurocholate (TC) and bromosulfophthalein (BSP). Uptake of MCs and cytotoxicity was demonstrated via MC-Western blot analysis, immunocytochemistry, cell viability and PP inhibition assays. All MC congeners bound covalently and inhibited mWBC PP. MC-LF was the most cytotoxic congener followed by -LW and -LR. The lowest toxin concentration significantly reducing mWBC viability after 48 h exposure was 400 nM (MC-LF). Uptake of MCs into mWBCs was inhibited via co-incubation with excess TC (50 and 500 microM) and BSP (50 microM). MC-Western blot analysis demonstrated a concentration-dependent accumulation of MCs. In conclusion, the in vitro data support the assumed MC-congener-dependent uptake in a mOatp-associated manner and cytotoxicity of MCs in primary murine whole brain cells.

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Comparison of Liquid Chromatography/Mass Spectrometry, ELISA, and Phosphatase Assay for the Determination of Microcystins in Blue-Green Algae Products

Cited by 83 publications

References 0 publications

Improving microcystin monitoring relevance in recreative waters: A regional case-study (Brittany, Western France, Europe)

Improving microcystin monitoring relevance in recreative waters: A regional case-study (Brittany, Western France, Europe)

Toxins of cyanobacteria

Oatp-associated uptake and toxicity of microcystins in primary murine whole brain cells

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