Does Your Dog Have Anxiety After a Rough Day at the Lake: Analysis of CBD Extracts for Dog Treats

Importance of the Topic

The rising legal acceptance of hemp-derived products has driven consumer interest in cannabidiol supplements for pets. Owners seek natural anxiolytic and anti-inflammatory remedies, yet variability in cultivation and extraction methods leads to inconsistent product quality and uncertain bioactive composition.

Objectives and Study Overview

This work applied an untargeted analytical approach using accurate-mass GC/Q-TOF with low-energy electron ionization to profile six commercial CBD oil pet supplements. Key aims included assessing chemical composition, detecting cannabinoids and related compounds, and comparing measured profiles to labeled CBD content.

Methodology

Samples from six brands were each prepared in five replicates by 500× dilution in dichloromethane and spiked with deuterated PAHs as internal standards. A n-alkane mixture provided retention index data. The injection sequence included blanks, pooled quality control samples, and randomized replicates. Data were acquired on a dual-column GC/Q-TOF platform at 70 eV and 14 eV to enhance molecular ion detection.

Used Instrumentation

• Gas Chromatograph: Agilent 7890B with split injection (10:1) and Ultra Inert single-taper wool liner
• Columns: Two × DB-35ms UI, 15 m length, 0.25 mm i.d., 0.25 µm film, connected via a purged union
• Mass Spectrometer: Agilent 7250 GC/Q-TOF with Source at 300 °C, Quadrupole at 150 °C
• Acquisition: Spectral range 45–650 m/z, 5 Hz rate, centroid and profile modes, EI at 70 eV and 14 eV
• Data Analysis: Agilent MassHunter Qualitative and Quantitative modules, Mass Profiler Professional, NIST17 library matching

Main Results and Discussion

Feature finding combined with library matching and accurate-mass confirmation enabled identification of over 840 entities across samples. Unsupervised PCA and hierarchical clustering revealed distinct chemotypes characterized by fatty acids, esters, triglycerides, tocopherols, sterols, and terpenoids. Low-energy EI at 14 eV preserved labile molecular ions, improving identification confidence for compounds such as α,β-amyrin, hentriacontane, and cannabidiol. Notably, dronabinol, a psychoactive THC isomer, was detected at significant levels in one extract. Quantitative comparison of relative peak areas showed that only one product matched its declared CBD content, while others deviated substantially or lacked disclosure.

Benefits and Practical Applications

• Comprehensive untargeted profiling for quality control and regulatory compliance of CBD pet supplements
• Enhanced detection of fragile bioactive molecules via low-energy electron ionization
• Robust data alignment and statistical analysis for batch consistency and counterfeit screening

Future Trends and Potential Applications

Integration of low-energy GC/Q-TOF workflows with tandem LC/MS techniques may further broaden molecular coverage. Standardization of pet supplement testing protocols and expanded libraries will support regulatory frameworks and consumer safety. Advances in data analytics and machine learning could enable rapid fingerprinting of complex botanicals.

Conclusion

The high-resolution accurate-mass GC/Q-TOF method with dual-energy electron ionization provided reliable untargeted characterization of CBD pet oil supplements. This approach delivered reproducible feature detection, improved molecular ion confirmation for labile compounds, and highlighted significant variability in product composition relative to labeling.

References

• Simão da Silva KAB, et al. Activation of cannabinoid receptors by the pentacyclic triterpene α,β-amyrin inhibits inflammatory and neuropathic persistent pain in mice. Pain. 2011;152(8):1872–1887.