Metabolomic Approach cum Advanced Mass Spectrometry Tools for Meat Differentiation of Zabeeha and Non-Zabeeha Slaugthered Animals

Prof. Dr. Syed Ghulam Musharraf

Metabolomic Approach cum Advanced Mass Spectrometry Tools for Meat
Differentiation of Zabeeha and Non-Zabeeha Slaugthered Animals:Chicken as a Case Study

Syed Ghulam Musharraf a,* , Nasir Abbasa, Arslan Alia, Adnan Husainb, M. Iqbal Choudharya and Atta-ur-Rahmana


In Muslim community, there is a major concern regarding Islamic and non-Islamic ways of
slaughtering animals, it is expected to have a good quality of meat, if it is slaughtered by
Islamic way in comparison with non-Islamic slaughtering. where the amount of blood left in the body that alters the physiochemical property of meat quality, by growing microbes and lipid oxidation due to iron (Fe) and cobalt (Co). Unfortunately, limited number of studies has been conducted to resolve various issues related to zabeeha (Islamic way of slaughtering) and non-zabeeha (non-Islamic way of slaughtering). However, differentiation of meet obtains from zabeeha and non-zabeeha slaughtering process is a major concern to the Muslim community.

Omics approach particularly use of metabolomics has a potential to be used for monitoring the food quality and processing. Similarly, advancement in mass spectrometry
offers a comprehensive solution for the analysis of complex biological samplers like meat in
a high-throughput manner with enhanced selectivity and sensitivity. In this study we have
considered chicken meat as it is the most common and cheap source of meat that is widely
being used worldwide, as a case study. Meat samples obtained from zabeeha and non-zabeeha slaughtering process were subjected to UPLC-HR-ESI-MS/MS analysis followed by
chemometric evaluation. A characteristic pattern was observed among the analysed samples.
Some peaks have shown an up-fold and down-fold changes between two groups and can be used for meat differentiation. Metabolites were identified based on various parameters including high-resolution masses, MS/MS analysis, database comparison etc. Outcome of this work will be highly beneficial for the food controlling Laboratories and Muslim Ummah,particularly.


Key Findings

Metabolomic profiling of chicken meat metabolites was performed in order to identify those compounds that can serve as markers to differentiate the slaughtering process of the chicken, i.e. if the chicken was slaughtered according to Islamic rights (Zabiha), and
non-Islamic way (Non Zabiha).
Several metabolites were found that can be used to differentiate between these groups. An extensive chemometric analysis was also performed. PCA plot has shown that the both groups lie separately and clustered individually (Figure 1). Similarly, score scatter plot of
PLSD was also used to evaluate the group clustering and found a clear separation among the group (Figure 2). Hierarchical clustering of the analyzed samples was performed to produce a dendrogram of all masses appeared in the spectra with different responses which illustrate similar types of metabolites among the two groups (Figure 3). Metabolites which are appeared in blue or red color shows a clear separation trend among the analyzed samples and therefore can be selected for further processing. However, most of the region was quite similar which means that majority metabolites are similar in both Zabiha and Non Zabiha. Some results were obtained from the Volcano plot (Figure 4).Progressions of those metabolites which have shown a characteristic pattern were further selected to monitor
their intensities in all analyzed samples. Three compounds were selected which have shown a up- or down-regulation profile. Two of them were identified based on their high resolution molecular masses,molecular formula, isotopic pattern and MS/MS features. Moreover, identification of compounds was further confirmed through the comparative analysis of their spectral data with the open database which includes Mass Spectral data and NIST data. Metabolite at m/z430 has shown an up regulation in Zabiha samples while metabolites at m/z 161 and m/z 418 have shown a down regulation pattern in Zabiha samples (Figure 5). In conclusion, the LC-ESI-MS/MSmethod can be used for the differentiation of meet samples obtained from Zabiha and Zabiha processes.


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