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Evaluation of immunoassay kits for determination of ochratoxin A in cereals4.1.4.3.2.1.2 LFDAs the LFD reader of Charm generates directly readings in concentration (ng/g), a different approach was used.Calibration curves using linear regression were constructed, the matrix effect was calculated by comparing theslopes 72 .Equation 16 CR = Slope [Diluted matrix] / Slope [Undiluted matrix] * 1004.1.4.4 Work package IV: Accuracy4.1.4.4.1 Study design4.1.4.4.2 Data handling and statistical analysesSamples are firstly analysed using the manufacturers protocol.The following protocol was used 73 .Equation 17xe-xt= (t*sd) / √NWith:x e: experimental mean (ng/g)x t: true value (ng/g)t: statistic value at 95% confidence level for levels of probabilityN: Number of replicate measurements4.1.4.5 Work package V: Blank sample analysis4.1.4.5.1 Study design3 extracts of blank matrices (wheat, oats, winter barley) are prepared according to the manufacturers guidelines. Onthree different days each extract was determined.4.1.4.5.2 Data handling and statistical analysesSamples are firstly analysed using the manufacturers protocol preferably at least three analyses per sample.Using a protocol described by reference 73 for estimation of detection limit.72Matuszewski BK. J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Jan 18;830(2):293-30073Skoog, West & Holler, Fundamentals of Analytical chemistry, 7 th Edition4-44 Experimental evaluation | CODA-CERVA
Evaluation of immunoassay kits for determination of ochratoxin A in cerealsEquation 18∆Xmin = xe-xt > (t*sd) * √(N1+N2)/(N1*N2)With:x e: experimental mean (ng/g)x t: true value (ng/g)sd: standard deviation (ng/g)t: statistic value at 95% confidence level for levels of probabilityN: Number of replicate measurements4.1.5 Extraction protocolsIn Table 43, the extraction protocols of the four kits are schematised.Table 43: Extraction protocolsCharm Sciences Europroxima R-Biopharm RomerlabsSample 10 g ≤sample≤ 50 g 2.5 g 5 g 20 gSolvent70 % MeOH2 (vol) on 1 (mass)1. H3P04 :10 ml+CH2Cl2 : 10 ml2. Evaporation3. Reconstitutionproprietary buffer4. Defatting hexane0.13 M NaHCO3100 ml70 % MeOH100 mlExtraction Shaking Shaking: Shaking Shaking/blendingExtractionTime2-3 min 20 min 15 min 3 minCODA-CERVA | Experimental evaluation 4-45
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CODA-CERVAEvaluation of immunoassay
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We would like to thank the kit manu
Page 8 and 9: 4.1.4.2 Work package II: Cross reac
Page 10 and 11: Table 45: Cross-reactivity in matri
Page 13: 1. Foreword & scope &background
Page 16 and 17: Evaluation of immunoassay kits for
Page 21: Evaluation of immunoassay kits for
Page 27: 3. Administrative evaluation
Page 37 and 38: Measured value (ng/g)Evaluation of
Page 51: 4. Experimental evaluation
Page 60 and 61: ()Evaluation of immunoassay kits fo
Page 62 and 63: Observed Concentration (ng/g)Observ
Page 79: 5. Conclusion and futureprospects
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Document - FAVV

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