Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins
A phosphatase inhibition assay for detection of okadaic acid (OA) toxins in shellfish, OkaTest, was single laboratory validated according to international recognized guidelines (AOAC, EURACHEM). Special emphasis was placed on the ruggedness of the method and stability of the components. All reagents...
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| Format: | Article |
| Language: | English |
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MDPI AG
2012-04-01
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| Series: | Toxins |
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| Online Access: | http://www.mdpi.com/2072-6651/4/5/339 |
| _version_ | 1828146398905761792 |
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| author | Luis Mata Elena Domínguez Pedro Razquin Dolores Calvo Henry G. F. Smienk |
| author_facet | Luis Mata Elena Domínguez Pedro Razquin Dolores Calvo Henry G. F. Smienk |
| author_sort | Luis Mata |
| collection | DOAJ |
| description | A phosphatase inhibition assay for detection of okadaic acid (OA) toxins in shellfish, OkaTest, was single laboratory validated according to international recognized guidelines (AOAC, EURACHEM). Special emphasis was placed on the ruggedness of the method and stability of the components. All reagents were stable for more than 6 months and the method was highly robust under normal laboratory conditions. The limit of detection and quantification were 44 and 56 µg/kg, respectively; both below the European legal limit of 160 µg/kg. The repeatability was evaluated with 2 naturally contaminated samples. The relative standard deviation (RSD) calculated was 1.4% at a level of 276 µg/kg and 3.9% at 124 µg/kg. Intermediate precision was estimated by testing 10 different samples (mussel and scallop) on three different days and ranged between 2.4 and 9.5%. The IC<sub>50</sub> values of the phosphatase used in this assay were determined for OA (1.2 nM), DTX-1 (1.6 nM) and DTX-2 (1.2 nM). The accuracy of the method was estimated by recovery testing for OA (mussel, 78–101%; king scallop, 98–114%), DTX-1 (king scallop, 79–102%) and DTX-2 (king scallop, 93%). Finally, the method was qualitatively compared to the mouse bioassay and LC-MS/MS. |
| first_indexed | 2024-04-11T20:46:50Z |
| format | Article |
| id | doaj.art-0e58ab5580e34bad9129e0761b0e81ad |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-11T20:46:50Z |
| publishDate | 2012-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-0e58ab5580e34bad9129e0761b0e81ad2022-12-22T04:04:00ZengMDPI AGToxins2072-66512012-04-014533935210.3390/toxins4050339Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid ToxinsLuis MataElena DomínguezPedro RazquinDolores CalvoHenry G. F. SmienkA phosphatase inhibition assay for detection of okadaic acid (OA) toxins in shellfish, OkaTest, was single laboratory validated according to international recognized guidelines (AOAC, EURACHEM). Special emphasis was placed on the ruggedness of the method and stability of the components. All reagents were stable for more than 6 months and the method was highly robust under normal laboratory conditions. The limit of detection and quantification were 44 and 56 µg/kg, respectively; both below the European legal limit of 160 µg/kg. The repeatability was evaluated with 2 naturally contaminated samples. The relative standard deviation (RSD) calculated was 1.4% at a level of 276 µg/kg and 3.9% at 124 µg/kg. Intermediate precision was estimated by testing 10 different samples (mussel and scallop) on three different days and ranged between 2.4 and 9.5%. The IC<sub>50</sub> values of the phosphatase used in this assay were determined for OA (1.2 nM), DTX-1 (1.6 nM) and DTX-2 (1.2 nM). The accuracy of the method was estimated by recovery testing for OA (mussel, 78–101%; king scallop, 98–114%), DTX-1 (king scallop, 79–102%) and DTX-2 (king scallop, 93%). Finally, the method was qualitatively compared to the mouse bioassay and LC-MS/MS.http://www.mdpi.com/2072-6651/4/5/339protein phosphatase inhibition assay (PPIA)protein phosphatase 2A (PP2A)validationokadaic acid (OA)diarrheic shellfish poisoning (DSP) |
| spellingShingle | Luis Mata Elena Domínguez Pedro Razquin Dolores Calvo Henry G. F. Smienk Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins Toxins protein phosphatase inhibition assay (PPIA) protein phosphatase 2A (PP2A) validation okadaic acid (OA) diarrheic shellfish poisoning (DSP) |
| title | Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins |
| title_full | Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins |
| title_fullStr | Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins |
| title_full_unstemmed | Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins |
| title_short | Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins |
| title_sort | single laboratory validation of a ready to use phosphatase inhibition assay for detection of okadaic acid toxins |
| topic | protein phosphatase inhibition assay (PPIA) protein phosphatase 2A (PP2A) validation okadaic acid (OA) diarrheic shellfish poisoning (DSP) |
| url | http://www.mdpi.com/2072-6651/4/5/339 |
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