LC/Q-TOF Analysis and Nontargeted Chemometric Profiling of Meats and Plant-Based Alternatives

Importance of the Topic

The growing demand for meat alternatives driven by health, environmental and regulatory factors requires objective analytical approaches to ensure that plant-based and cell-based products match the sensory attributes of conventional meats. Traditional sensory testing is subjective, so high-resolution nontargeted liquid chromatography–mass spectrometry (LC–MS) combined with chemometric analysis offers an unbiased solution for profiling flavor compounds and guiding product development.

Objectives and Study Overview

This study presents a comprehensive, nontargeted profiling workflow using an Agilent 1290 Infinity II LC coupled to a 6546 Q-TOF MS to characterize chemical components in real meats (chicken, beef, pork) and their commercial plant-based analogs. Advanced statistical tools translate complex accurate-mass data into clear information on compound identities, abundance patterns and their correlation with target taste profiles.

Methodology and Instrumentation

Sample Preparation:

• Weighed meat or plant-based samples in polypropylene tubes.
• Extracted twice with 70:30 methanol:water (v/v), vortexed 10 min, centrifuged 4 000 rpm for 15 min.
• Filtered extracts through 0.45 µm PES filters into amber autosampler vials.

Instrumentation:

• LC: Agilent 1290 Infinity II with high-speed pump, multisampler, multicolumn thermostat.
• Column: Poroshell 120 EC-C18 (3.0×100 mm, 2.7 µm).
• MS: Agilent 6546 Q-TOF operated in positive All Ions fragmentation mode (m/z 100–1 700; collision energies 10, 20, 40 V).
• Mobile phases: 10 mM ammonium formate/0.1% formic acid in water and acetonitrile.
• Data processing: MassHunter Qualitative 10.0, Profinder 10.0, Mass Profiler Professional 15.1.

Key Results and Discussion

• PCA score plots showed tight clustering of triplicate injections, confirming method repeatability. Meat types and their plant-based equivalents formed distinct groups aligned with target flavor categories.
• Loading plots identified specific compounds driving differences: amino acids, short peptides and nucleotides linked to umami and characteristic grilled-meat flavors.
• Heat maps of amino acid abundances revealed higher levels of bitter amino acids in plant-based beef analogs (PBB 3, 4) compared to real beef and diverse patterns of nucleotides and peptides in plant-based chicken (PBC 1, 2).
• Lower purine-derived hypoxanthine in PBC 2 suggests potential health benefits by reducing uric acid formation while affecting umami intensity.

Benefits and Practical Applications

• Provides objective, untargeted detection of known and unknown flavor compounds across broad m/z ranges.
• Enables rapid comparison of commercial and novel formulations to benchmark sensory profiles.
• Guides ingredient selection and process adjustments to replicate taste, texture and nutritional attributes of animal meats.
• Supports quality control by tracking batch-to-batch consistency and identifying off-profile compounds.

Future Trends and Opportunities

• Integration with machine learning algorithms for automated pattern recognition and predictive flavor modeling.
• Expansion of custom fragmentation libraries to include emerging food ingredients and bioactive compounds.
• Real-time, in-line LC–MS monitoring in production environments for dynamic process control.
• Application to cell-cultured meats and hybrid protein products to accelerate product innovation.

Conclusion

The described nontargeted All Ions LC–QTOF workflow combined with chemometric and visualization tools offers a robust strategy for profiling complex flavor compounds in meats and their plant-based analogs. This approach delivers actionable molecular insights that support sensory optimization, quality assurance and accelerated development of alternative protein foods.

Reference

1. Kaczmarska K, et al. Flavor and Metabolite Profiles of Meat, Meat Substitutes, and Traditional Plant-Based High-Protein Food Products Available in Australia. Foods. 2021;10(4):801.
2. Ueda Y, et al. Flavor Characteristics of Glutathione in Raw and Cooked Foodstuffs. Biosci. Biotechnol. Biochem. 1997;61(12):1977–1980.
3. Jakse B, et al. Uric Acid and Plant-Based Nutrition. Nutrients. 2019;11(8):1736.