<a class="affpopup" href="https://www.ncbi.nlm.nih.gov/pubmed/?term=Hameed%20AM%5BAuthor%5D&amp;cauthor=true&amp;cauthor_uid=30112151">Ademola Monsur Hameed</a>,<sup>1,</sup><sup>*</sup> <a class="affpopup" href="https://www.ncbi.nlm.nih.gov/pubmed/?term=Asiyanbi-H%20T%5BAuthor%5D&amp;cauthor=true&amp;cauthor_uid=30112151">Tawakalit Asiyanbi-H</a>,<sup>2</sup> <a class="affpopup" href="https://www.ncbi.nlm.nih.gov/pubmed/?term=Idris%20M%5BAuthor%5D&amp;cauthor=true&amp;cauthor_uid=30112151">Munirat Idris</a>,<sup>1</sup> <a class="affpopup" href="https://www.ncbi.nlm.nih.gov/pubmed/?term=Fadzillah%20N%5BAuthor%5D&amp;cauthor=true&amp;cauthor_uid=30112151">Nurrulhidayah Fadzillah</a>,<sup>1</sup> and <a class="affpopup" href="https://www.ncbi.nlm.nih.gov/pubmed/?term=Mirghani%20ME%5BAuthor%5D&amp;cauthor=true&amp;cauthor_uid=30112151">Mohamed Elwathig S. Mirghani</a><sup>1</sup>
<h2 id="idm140686122385024title" class="head no_bottom_margin ui-helper-clearfix">Abstract</h2>
<div>
<p id="__p1" class="p p-first-last">Gelatin Source is a very popular pharmaceutical and food ingredient and the most studied ingredient in <a href="https://halalnews.info/">Halal researches</a>. Interest in source gelatin authentication is based on religious and cultural beliefs, food fraud prevention and health issues. Seven gelatin authentication methods that have been developed include: nucleic acid based, immunochemical, electrophoretic analysis, spectroscopic, mass-spectrometric, chromatographic-chemometric and chemisorption methods. These methods are time consuming, and require capital intensive equipment with huge running cost. Reliability of gelatin authentication methods is challenged mostly by transformation of gelatin during processing and close similarities among gelatin structures. This review concisely presents findings and challenges in this research area and suggests needs for more researches on development of rapid authentication method and process-transformed gelatins.</p>

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<h2 id="__sec1title" class="head no_bottom_margin ui-helper-clearfix">INTRODUCTION</h2>
<p id="__p3" class="p p-first">As at 2013, the global Halal market size was about 580 to 660 billion US Dollars and has great potential to increase in the future year (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b15-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Jahangir <em>et al.</em> 2016</a>). Apart from increasing Muslim population and Halal awareness, Halal market is driven by development of Halal ingredients authentication methods that strengthening consumers’ confident (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b20-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Malik <em>et al.</em> 2016</a>). Gelatin is among the most studied Halal ingredient because of its vast usage in pharmaceutical and food products. Gelatin is a hydrocolloid with unique properties and can function as gelling, thickener, foaming agent, plasticizer, texture and binding agent (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b27-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Sahilah <em>et al.</em> 2012</a>).</p>
<p id="__p4">The sources of most commercial. Gelatin are from mammalian (bovine and mostly porcine). Bone and hide (<a id="__tag_716050795" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b30-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Shabani <em>et al.</em> 2015</a>). Consumers’ concern about gelatin is due to cultural and religious beliefs. For examples, Muslims and Jews reject porcine based food derivatives, Chinese traditional medicine use only gelatin from donkey skin for treatment of some disease (<a id="__tag_716050794" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b23-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Nemati <em>et al.</em> 2004</a>) and vegetarians avoid animal based products. Increase in Halal awareness among Muslims has called for great need of food-source authentications. Gelatin and gelatin based products are currently classified as doubtful because haram (porcine) gelatin is the most abundant. Traceability of gelatin has been a great task in Halal field. Other reasons for gelatin source authentication include: (i) protection of consumer from food frauds due to false labeling and avoid unfair market competition (<a id="__tag_716050783" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b17-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Kesmen <em>et al.</em> 2009</a>) and (ii) health issues. The use of bovine carrying prion proteins has been linked to bovine spongiform encephalopathy (mad cow disease) causing immune response that led to fatal neurodegenerative disease (spongy degeneration of the brain and spinal cord) in cattle (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b37-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Widyaninggar <em>et al.</em> 2012</a>). Also, bovine and porcine gelatins is risky to gelatin-allergic patients (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b26-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Raraswati <em>et al.</em> 2013</a>).</p>

<h2 id="__p5">Gelatin contains protein</h2>
peptides or nucleic acids (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b20-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Malik <em>et al.</em> 2016</a>). Proteins, peptide derivatives and nucleic acids are favorable means for tissue identification in samples even after it has undergone heating, for instance, in processed food (<a id="__tag_716050788" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b6-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Buckley <em>et al.</em> 2009</a>). However, large similarities between structures of gelatin from different sources make their differentiation difficult (<a id="__tag_716050799" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b23-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Nemati <em>et al.</em> 2004</a>). Many gelatin source authentication methods have been developed including: spectroscopic method (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b12-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Hashim <em>et al.</em> 2010</a>; <a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b13-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Hermanto &amp; Fatimah 2013</a>), immunochemical method (<a id="__tag_716378676" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b33-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Tukiran <em>et al.</em> 2015</a>, <a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b34-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">2016a</a>), nucleic acid based method (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b7-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Cai <em>et al.</em> 2012</a>; <a id="__tag_716050807" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b8-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Demirhan <em>et al.</em> 2012</a>; <a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b20-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Malik <em>et al.</em> 2016</a>; <a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b22-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Mutalib <em>et al.</em> 2015</a>; <a id="__tag_716050776" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b30-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Shabani <em>et al.</em> 2015</a>; <a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b31-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Sudjadi <em>et al.</em> 2015</a>), mass spectrometric method HPLC/MS (<a id="__tag_716050778" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b39-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Yilmaz <em>et al.</em> 2013</a>; <a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b40-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">G.-F. Zhang <em>et al.</em> 2008</a>; <a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b41-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">G. Zhang <em>et al.</em> 2009</a>), chromatographic-chemometric method (<a id="__tag_716050805" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b3-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Azilawati <em>et al.</em> 2015</a>), electrophoretic analysis (<a id="__tag_716050781" class=" bibr popnode tag_hotlink tag_tooltip" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b4-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Azira <em>et al.</em> 2014</a>) and chemisorption (<a class=" bibr popnode" role="button" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/#b14-tlsr-29-2-213" aria-expanded="false" aria-haspopup="true">Hidaka &amp; Liu 2003</a>).
<p id="__p6" class="p p-last">We have reviewed gelatin source authentication methods and identified their achievements and challenges. Current gelatin source authentication methods are time consuming and expensive, thus required development of cheap and quick analytical methods. Likewise, gelatin transformation during processing can potentially hinder the efficiency of the currently developed gelatin authentication methods. This work will serve as a quick reference guide for future researchers.</p>

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<h2 id="__sec2title" class="head no_bottom_margin ui-helper-clearfix">AUTHENTICATION OF GELATIN SOURCES</h2>
<p id="__p7" class="p p-first">As earlier stated, several methods have been developed to investigate sources of gelatin in commercial gelatin or gelatin-containing products. Authentication of gelatin sources in several products have been investigated (<a class="fig-table-link figpopup" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072727/table/t1-tlsr-29-2-213/" target="table" rel="noopener noreferrer">Table 1</a>). Researchers have adopted several approaches to develop specie-specific gelatin authentication methods. The approaches include use of the amino acid profile, the presence of specific peptide and DNA markers and the spectral fingerprint of gelatins or its peptides.</p>

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