Detection and Differentiation of Bovine and Porcine Gelatins in Food and Pharmaceutical Products By LC/MS/MS Method
Applications | 2017 | ShimadzuInstrumentation
Gelatin, a polypeptide mixture from collagen hydrolysis, is extensively used in food pharmaceutical and cosmetic sectors. Determining its animal origin is vital for religious dietary compliance quality control and safety assessment due to zoonotic risk concerns.
This work presents the development of a targeted proteomic method using LC/MS/MS to identify species specific peptide markers differentiating bovine and porcine gelatins. Over forty candidate peptides were screened to establish sensitive robust markers applicable to commercial samples.
Sample preparation involved aqueous extraction digestion with trypsin and direct analysis without solid phase extraction. Centrifugation separated peptides prior to injection. UHPLC separation was performed on a reverse phase peptide column using formic acid in water and acetonitrile gradient. MS/MS detection used multiple reaction monitoring in positive mode with optimized transition settings.
Nine bovine and eight porcine specific peptides were validated with signal to noise ratio above 3 and repeatability below 15 percent RSD. Analysis of commercial food and pharmaceutical samples successfully identified gelatin sources in seven of twelve products. Bovine markers showed higher sensitivity while four key porcine markers provided reliable detection. Mixed gelatin presence was demonstrated in one pharmaceutical capsule indicating method applicability to complex matrices.
The method reduces sample preparation steps and time compared to conventional assays. High sensitivity and multiple confirmation ions ensure confident authentication of gelatin origin supporting halal kosher and other dietary regulations as well as quality assurance in industrial laboratories.
Further work may focus on quantitative calibration to measure gelatin ratios in mixtures and extend marker libraries to other animal sources. Automation and integration with high throughput platforms will enhance routine screening in food and pharmaceutical industries.
A streamlined LC/MS/MS approach with validated species specific peptides provides a reliable tool for bovine porcine gelatin authentication in processed products. The protocol offers rapid high sensitivity analysis facilitating regulatory compliance and consumer protection.
LCMS 8060 triple quadrupole MS
UHPLC system with peptide XB C18 column
Heated electrospray ionization source
Multiple reaction monitoring mode
LC/MS, LC/MS/MS, LC/QQQ
IndustriesFood & Agriculture, Pharma & Biopharma
ManufacturerShimadzu
Summary
Significance of the Topic
Gelatin, a polypeptide mixture from collagen hydrolysis, is extensively used in food pharmaceutical and cosmetic sectors. Determining its animal origin is vital for religious dietary compliance quality control and safety assessment due to zoonotic risk concerns.
Objectives and Study Overview
This work presents the development of a targeted proteomic method using LC/MS/MS to identify species specific peptide markers differentiating bovine and porcine gelatins. Over forty candidate peptides were screened to establish sensitive robust markers applicable to commercial samples.
Methodology and Instrumentation
Sample preparation involved aqueous extraction digestion with trypsin and direct analysis without solid phase extraction. Centrifugation separated peptides prior to injection. UHPLC separation was performed on a reverse phase peptide column using formic acid in water and acetonitrile gradient. MS/MS detection used multiple reaction monitoring in positive mode with optimized transition settings.
Main Results and Discussion
Nine bovine and eight porcine specific peptides were validated with signal to noise ratio above 3 and repeatability below 15 percent RSD. Analysis of commercial food and pharmaceutical samples successfully identified gelatin sources in seven of twelve products. Bovine markers showed higher sensitivity while four key porcine markers provided reliable detection. Mixed gelatin presence was demonstrated in one pharmaceutical capsule indicating method applicability to complex matrices.
Benefits and Practical Applications
The method reduces sample preparation steps and time compared to conventional assays. High sensitivity and multiple confirmation ions ensure confident authentication of gelatin origin supporting halal kosher and other dietary regulations as well as quality assurance in industrial laboratories.
Future Trends and Potential Applications
Further work may focus on quantitative calibration to measure gelatin ratios in mixtures and extend marker libraries to other animal sources. Automation and integration with high throughput platforms will enhance routine screening in food and pharmaceutical industries.
Conclusion
A streamlined LC/MS/MS approach with validated species specific peptides provides a reliable tool for bovine porcine gelatin authentication in processed products. The protocol offers rapid high sensitivity analysis facilitating regulatory compliance and consumer protection.
Used Instrumentation
LCMS 8060 triple quadrupole MS
UHPLC system with peptide XB C18 column
Heated electrospray ionization source
Multiple reaction monitoring mode
References
- H H Grundy P Reece M Buckley C M Solazzo A A Dowle D Ashford A J Charlton M K Wadsley M J Collins Food Chem 2016 190 276 284
- M E Ali S Sultana S B A Hamid M A M Hossain W A Yehya M A Kader S K Bhargava Crit Rev Food Sci Nutr 2016 56 1 17
- U Jumhawan J Xing Z Zhan Application News AD 0153 Shimadzu 2017
- M Nemati M R Oveisi H Abdollahi O Sabzevari J Pharm Biomed Anal 2004 34 485 492
- G Zhang T Liu Q Wang L Chen J Lei J Luo G Ma Z Su Food Hydrocolloids 2009 23 2001 2007
- X Cheng F Wei X Xiao Y Zhao Y Shi W Liu P Zhang S Ma S Tian R Lin J Pharm Biomed Anal 2012 62 191 195
- C Flaudrops N Armstrong D Raoult E Chabriere J Food Comp Anal 2015 41 104 112
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