Gas Chromatography Mass Spectrometry Analysis Reveals the Differences in Volatile Metabolites of Royal Jelly from Different Honeybee Stocks

Significance of the Topic

Royal jelly plays a pivotal role in honeybee caste determination and longevity of queen bees. Its unique volatile composition contributes to product aroma and consumer acceptance. Despite its importance, comparative data on volatile metabolites from different bee stocks remain scarce.

Objectives and Study Overview

This study aimed to characterize and compare volatile profiles of royal jelly produced by three honeybee stocks: high royal jelly–producing bees (RJBs), Italian bees (ITBs), and American Italian bees (AITBs). Triplicate royal jelly samples were collected under identical apiary conditions and stored at –80 °C prior to analysis.

Methodology and Instrumentation

Volatile compounds were extracted by headspace-solid phase microextraction (HS-SPME). Analysis employed an Agilent 7890B gas chromatograph coupled to a 7000D triple quadrupole mass spectrometer in full-scan mode. Data processing included MassHunter Qualitative deconvolution and Mass Profiler Professional for frequency and variability filtering, ANOVA, principal component analysis (PCA), and hierarchical clustering.

Main Results and Discussion

Out of 121 differential entities, 37 volatile metabolites were identified, comprising aldehydes, esters, alkanes, alcohols, phenols, ketones, and others. PCA accounted for 91.33% of variance in the first three components, clearly separating RJB samples from ITB and AITB. Hierarchical clustering grouped ITB and AITB together, with RJB forming a distinct cluster, indicating characteristic volatile signatures. RJB samples contained unique guaiazulene and higher levels of sweet/floral volatiles, while ITB samples were enriched in phenolic compounds (e.g., 2-methylphenol, 2-methoxyphenol) and AITB shared indole with ITB.

Benefits and Practical Applications

• Volatile markers enable authentication and quality control of royal jelly products.
• Data support selective breeding of bee stocks for desirable aroma profiles.
• Chemometric workflows facilitate rapid discrimination of RJ origin.

Future Trends and Applications

• Integration of sensory evaluation with metabolomic profiling to link volatile composition to flavor perception.
• Development of portable sensors targeting key volatiles for on-site quality assessment.
• Investigation of environmental and genetic factors shaping volatile metabolomes in royal jelly.

Conclusion

This work delivers the first comparative volatile metabolite profiling of royal jelly from high-yield and conventional bee stocks, identifying 37 signature compounds that differentiate RJ origins and influence aroma quality. The findings enhance biochemical understanding and pave the way for improved quality control and bee breeding strategies.

Reference

1. Patel N.G., Haydak M.H., Gochnauer T.A. Electrophoretic Components of the Proteins in Honeybee Larval Food. Nature. 1960;186:633.
2. Lensky Y., Rakover Y. Separate protein body compartments of the worker honeybee (Apis mellifera L.). Comp Biochem Physiol B. 1983;75:607–615.