A Complete Solution for Gelatin Species Authentication in Routine Analysis Using ProteinWorks Auto-eXpress Digest Kit and Xevo TQ-XS

Significance of the Topic

Gelatin is a versatile biopolymer widely applied in food, pharmaceutical and cosmetic industries. Establishing the animal origin of gelatin is critical for public health, regulatory compliance and religious requirements, particularly halal certification. Accurate species authentication protects consumers and manufacturers from adulteration and labeling errors.

Study Objectives and Overview

This study presents a streamlined protocol for gelatin species authentication combining a three-step sample preparation using the ProteinWorks Auto-eXpress Digest Kit with a sensitive LC-MS/MS analysis on the Xevo TQ-XS system. The method aims to detect bovine and porcine gelatin in routine testing workflows within a single day.

Methodology and Instrumentation

Samples and reference standards were pretreated with ammonium bicarbonate, then digested in three hours using the ProteinWorks Auto-eXpress Low 3 Digest Kit. Peptide separation was achieved on an ACQUITY UPLC I-Class system with an ACQUITY Premier UPLC HSS T3 column at 40 °C. Tandem mass spectrometry detection was performed on a Xevo TQ-XS in positive ESI mode, monitoring multiple MRM transitions for nine species-specific peptides. Data acquisition and processing used MassLynx and TargetLynx software.

Results and Discussion

The optimized protocol enabled complete digestion and analysis within three hours. Nine peptide markers (five bovine, four porcine) were monitored with 4–5 MRM transitions each. The method achieved reliable identification using criteria of at least three transitions per peptide and two peptides per species. Analysis of eight commercial candies correctly distinguished halal-certified (bovine gelatin) and non-halal (porcine gelatin) samples. Adulteration experiments demonstrated detection of 1 % porcine gelatin in bovine matrix.

Benefits and Practical Applications

• Same-day sample preparation and analysis.
• Simultaneous detection of multiple animal sources.
• High sensitivity and selectivity at low adulteration levels.
• Suitable for routine quality control in food and pharmaceutical industries.

Future Trends and Applications

Future developments may extend the marker panel to include fish and poultry gelatin, implement full automation for high-throughput screening, and integrate authentication data into compliance databases. Emerging MS technologies and bioinformatics tools will further enhance speed and robustness.

Conclusion

The presented workflow offers a robust, rapid and scalable solution for routine gelatin species authentication, meeting the needs of regulatory and industry laboratories for accurate labeling and consumer protection.

References

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