Abraxis Test Method 522015


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Analytical testing dots

Source:
Abraxis

Method Name:
Microcystins in water by Immunoassay, Direct Monoclonal Microtiter Plate. Official Name: Abraxis Microcystin-DM ELISA Plate Kit PN 522015

Description:
Microcystins are detected using a colorimetric immunoassay (ELISA) procedure. A sample (0.10 mL), enzyme conjugate (Microcystins-HRP), and an antibody solution against Microcystins are added to plate wells pre-coated with monoclonal antibodies (goat anti-mouse). Both the Microcystins (if present) in the sample and Microcystins-HRP conjugate compete to bind to the antibody solution in proportion to their concentrations. The anti-Microcystins antibodies are then bound to the immobilized monoclonal antibodies on the plate. After an incubation, the unbound molecules are washed and decanted. A substrate is then added which is catalyzed by the enzyme and converted from a colorless to a blue solution. The reaction is terminated with the addition of a dilute acid. The concentration of Microcystins in the sample is determined by measuring its absorbance at a specific wavelength (450nm) using a photometer and comparing its absorbance to the absorbance of the calibrators.

Scope:
This method determines Microcystins and Nodularins in water (groundwater, surface water, well water).

Citation:
Abraxis User Guides and Flowcharts

Interferences:
Cross-reactivity: Microcystins (YR, RR, LA) and Nodularins, and high concentrations of N-hemi-ADDA and ADDA may produce false positive responses for Microcystin-LR.

QC Requirements:
(A) Calibration with 5 standards and 1 blank, all analyzed in duplicate. (B) Precision: 3 matrix samples with different levels in the range for quantitative analysis analyzed daily for 5 days with 5 replicates in each of 5 assays. (C) Accuracy: 5 matrix samples spiked with the target analyte at 4 different levels in the range for quantitative analysis. (D) Validation: Analysis of 4 positive and 4 negative samples by an independent method, for confirmation.

Maximum Holding Time:
14 days at 4 C, longer if frozen

Media:
WATER

Subcategory:
Organic

Concentration:
0.10 - 5.0

Precision:
-

Detection:
-

Revision Number:
2007

Analytical testing dots

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Analytical testing dots