**3. Results and discussion**

The results of this study were comprehensively described in **Tables 3** and **4**. **Figure 1** showed the linearity curve of AFM1 standard concentrations of 0.05, 0.1, 0.2, 0.3, 0.4 and 0.5 μg/ml. The method showed linear response R<sup>2</sup> = 1.

**Table 3** shows the level of AFM1 of all dairy products that exceeds the tolerable limits (0.050 μg/kg) established by European Commission Regulation.

Our results concluded that 33.3% of processed butter sample showed positive recovery of AFM1 with range concentration above EU limits (0.050 μg/kg) while no local butter sample with AFM1 toxicity was found (**Table 3**). These results are only in agreement with a study conducted by Fallah et al. [16] who analyzed 31 butter samples and got 25.8% AFM1-positive ones with range above permissible limits established by EU.

The similar trend was found in case of processed cheese samples which were found contaminated (33.3%) with AFM1 (**Table 3**) whereas in another study the AFM1 concentration was found much higher i.e., 78% [15]. There are many other such studies having positive percentages of AFM1 higher than our results [16–18].

Similarly, analysis of processed and unprocessed yogurt samples showed that 26.6% of former were contaminated with AFM1 above EU permissible limit whereas no sample in latter was found affected with these aflatoxins. Our results showed low positive percentage than documented by Iqbal and Asi [15] who found *An Assessment and Control of AFM1 in Milk and Main Dairy Products in Lahore, Pakistan DOI: http://dx.doi.org/10.5772/intechopen.99184*


### **Table 3.**

*HPLC results of aflatoxin M1 in dairy products.*


### **Table 4.**

*Efficacy of different toxin binders in groups A, B, and C.*

59 AFM1-positive samples. Many other such studies also showed higher incidents of AFM1 in yogurt samples than our study [19–21]. Lastly, results of unprocessed cream samples showed the highest concentration of AFM1 whereas no aflatoxin

**Figure 1.**

*Linearity curve of aflatoxin M1 standard concentrations.*

**Figure 2.** *Positive percentage in each group after offering toxin binders.*

was detected in processed cream samples as shown in **Table 3**. These results have not only local impacts but high impact at global level too. Because at global level, particularly in underdeveloped and developing countries, the topic of aflatoxins in dairy sector impacting humans as well as animals is a neglected topic.

During the second phase of the study, milk samples collected on 2nd, 3rd, 4th, and 7th day showed significantly different efficacies of three toxin binders in groups A, B, and C. It was found that toxin binder used in group C had significantly higher (<0.05) efficacy as compared to those used in groups A and B. This toxin binder C had yeast wall (75%) in combination with algae (25%) so it showed best results and eradicated AFM1 from all the animals after 48 h. The reason behind is the main role of yeast wall in the whole yeast which binds with mycotoxin and its binding ability is catalyzed by algae so the product having this combination provided the best results. Whereas, the clay-based toxin binder used in group A showed comparatively worst results in controlling AFM1 due to their lack of binding with these mycotoxins as described by Chestnut et al. [22]. Other disadvantages associated with clay-based toxin binders are their probable interaction with the essential nutrients [23] and high inclusion rates [24]. On the other hand, whole yeast-based toxin binder showed significantly lower efficacy as compared to group C and higher

*An Assessment and Control of AFM1 in Milk and Main Dairy Products in Lahore, Pakistan DOI: http://dx.doi.org/10.5772/intechopen.99184*

efficacy as compared to group A. The reason behind would be that whole yeast alone does not have good binding ability with mycotoxins so it also failed to control AFM1, comparatively. The whole findings are summarized in **Figure 2**.
