Determination of Animal Species Origin from Gelatin in Food and Pharmaceutical Products by LC-MS/MS

Significance of the Topic

Gelatin is widely used in food and pharmaceutical industries for its gelling and thickening properties.
Species origin verification is critical to address religious dietary restrictions (halal, kosher), prevent adulteration and mitigate risks such as BSE contamination.

Objectives and Study Overview

The study aimed to develop and validate a fast, robust and reliable LC-MS/MS method to determine bovine and porcine gelatin in commercial products.
Key goals included achieving a limit of quantitation of 1% w/w and demonstrating applicability across raw and processed samples.

Methodology and Instrumentation

Sample Preparation:

• 5 mg of gelatin or product dissolved in 50 mM ammonium bicarbonate at 37°C to 1 mg/mL.
• Enzymatic digestion conducted overnight at 37°C (10–15 h) or by a 5 min microwave burst.

LC Conditions:

• SCIEX ExionLC™ system with Phenomenex Kinetex® C18 column (50×3 mm, 2.6 μm) at 40°C.
• Gradient of 0.1% formic acid in water (eluent A) and acetonitrile (eluent B) over 19 min, flow rate 250 μL/min, injection volume 20 μL.

MS/MS Conditions:

• SCIEX QTRAP™ 4500 with Turbo V™ ESI source.
• Multiple Reaction Monitoring (MRM) using three transitions per species.

Main Results and Discussion

• System stability was confirmed over 3 days at 10°C with 10% gelatin samples.
• Limits of quantitation reached 1% w/w for both bovine and porcine gelatin.
• Microwave and overnight digestions provided equivalent detection of contaminations.
• Screening of 14 commercial products (raw gelatin, dairy, candies, sausages, capsules) accurately identified species without false positives in halal-certified samples.

Benefits and Practical Applications

• Fast analysis (19 min per run) and simple operation suited for routine testing.
• High selectivity and sensitivity via targeted MRM on a triple quadrupole instrument.
• Supports quality assurance in food and pharmaceutical sectors, ensuring compliance with religious and safety standards.

Future Trends and Applications

• Integration with high-resolution and multiplexed MS workflows to expand the range of detectable species.
• Automation and miniaturization of sample preparation and LC-MS platforms for on-site monitoring.
• Development of comprehensive peptide marker libraries and AI-driven data analysis for rapid authentication.

Conclusion

The validated LC-MS/MS method offers a rapid, robust and reliable approach to identify bovine and porcine gelatin at low contamination levels, facilitating routine screening in food and pharmaceutical products.

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