2.5.4. Calculations
Calculations were made according to the following equation:
Wm ¼ Wa
Vf
.
Vi
ð1=VsÞ (4)
where Wm ¼ amount of AFM1 in the test sample in ppb (mg/l);
Wa ¼ amount of AFM1 corresponding to area of AFM1 peak of the
test extract (ng); Vf ¼ the final volume of re-dissolved eluate (ml);
Vi ¼ volume of injected eluate (ml); and Vs ¼ volume of test portion
(centrifuged and filtrated Doogh) passing through the immunoaffinity
column (mL).
Total AFM1 binding (%) ¼ 100 (0.500 ppb Wm (ppb))/
0.500 ppb.
Probiotic affected AFM1 binding (%) ¼ Total AFM1 binding (%) in
probiotic treatment blank treatment.
2.5.5. Determination of performance characteristics
The limit of detection (LOD) (Valc
arcel, 2012) was 0.013 ng
AFM1/mL. To determine the recovery of AFM1, three blank Doogh
samples were contaminated at the rate of 0.050 ng AFM1/mL and
were prepared for analysis as described above. Their AFM1 contents
were calculated, and recovery of AFM1 was found to be between 77
and 86% (mean 81%). Therefore, the method was determined to be
accurate enough for use in the survey.
2.6. Statistical analysis
All assays in this study were carried out in triplicate. All data
were subjected to one-way analysis of variance (ANOVA), and the
significance of the difference between means was determined by
Duncan’s multiple range test (p < 0.05) using SPSS statistical software
(version 16; SPSS Inc., USA).
15
2.5.4. CalculationsCalculations were made according to the following equation:Wm ¼ Wa Vf.Vi ð1=VsÞ (4)where Wm ¼ amount of AFM1 in the test sample in ppb (mg/l);Wa ¼ amount of AFM1 corresponding to area of AFM1 peak of thetest extract (ng); Vf ¼ the final volume of re-dissolved eluate (ml);Vi ¼ volume of injected eluate (ml); and Vs ¼ volume of test portion(centrifuged and filtrated Doogh) passing through the immunoaffinitycolumn (mL).Total AFM1 binding (%) ¼ 100 (0.500 ppb Wm (ppb))/0.500 ppb.Probiotic affected AFM1 binding (%) ¼ Total AFM1 binding (%) inprobiotic treatment blank treatment.2.5.5. Determination of performance characteristicsThe limit of detection (LOD) (Valcarcel, 2012) was 0.013 ngAFM1/mL. To determine the recovery of AFM1, three blank Dooghsamples were contaminated at the rate of 0.050 ng AFM1/mL andwere prepared for analysis as described above. Their AFM1 contentswere calculated, and recovery of AFM1 was found to be between 77and 86% (mean 81%). Therefore, the method was determined to beaccurate enough for use in the survey.2.6. Statistical analysisAll assays in this study were carried out in triplicate. All datawere subjected to one-way analysis of variance (ANOVA), and thesignificance of the difference between means was determined byDuncan’s multiple range test (p < 0.05) using SPSS statistical software(version 16; SPSS Inc., USA).15
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