Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins

Toxins

Henry G. F. Smienk²  Dolores Calvo²  Pedro Razquin²  Elena Domínguez¹ 

[1] Zeu-Inmunotec, Polígono PLAZA, C/Bari 25 Dpdo, 50197, Zaragoza, Spain;

关键词: protein phosphatase inhibition assay (PPIA);    protein phosphatase 2A (PP2A);    validation;    okadaic acid (OA);    diarrheic shellfish poisoning (DSP);   

DOI  :  10.3390/toxins4050339

来源: mdpi

【 摘 要 】

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 IC₅₀ 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.

【 授权许可】

CC BY   
© 2012 by the authors; licensee MDPI, Basel, Switzerland.