Evaluation of fast enantioselective multidimensional gas chromatography methods for monoterpenic compounds: Authenticity control of Australian tea tree oil

• Fast GC–eGC and eGC × GC separation of target terpenes in tea tree oil is reported.
• Adequate compound separation from interferences allows FID detection to be used.
• GC–eGC and eGC × GC analyses are completed in 25 and < 20 min, respectively.
• Results for enantiomer compositions indicate acceptable quantitative measurement.
• Exact target chiral fingerprint patterns of pure Australian TTO are proposed.

This work demonstrates the potential of fast multiple heart-cut enantioselective multidimensional gas chromatography (GC − eGC) and enantioselective comprehensive two-dimensional gas chromatography (eGC × GC), to perform the stereoisomeric analysis of three key chiral monoterpenes (limonene, terpinen-4-ol and α-terpineol) present in tea tree oil (TTO). In GC − eGC, separation was conducted using a combination of mid-polar first dimension (1D) column and a chiral second dimension (2D) column, providing interference-free enantioresolution of the individual antipodes of each optically active component. A combination of 1D chiral column and 2D polar columns (ionic liquid and wax phases) were tested for the eGC × GC study. Quantification was proposed based on summation of two major modulated peaks for each antipode, displaying comparable results with those derived from GC − eGC. Fast chiral separations were achieved within 25 min for GC − eGC and < 20 min for eGC × GC, while ensuring adequate interference-free enantiomer separation. The suitability of using these two enantioselective multidimensional approaches for the routine assessment of chiral monoterpenes in TTO was evaluated and discussed. Exact enantiomeric composition of chiral markers for authentic TTOs was proposed by analysing a representative number of pure TTOs sourced directly from plantations of known provenance in Australia. Consistent enantiomeric fractions of 61.6 ± 1.5% (+):38.4 ± 1.5% (−) for limonene, 61.7 ± 1.6% (+):38.3 ± 1.6% (−) for terpinen-4-ol and 79.6 ± 1.4% (+):20.4 ± 1.4% (−) for α-terpineol were obtained for the 57 authentic Australian TTOs. The results were compared (using principle component analysis) with commercial TTOs (declared as derived from Melaleuca alternifolia) obtained from different continents. Assessing these data to determine adulteration, or additives that affect the enantiomeric ratios, in commercially sourced TTOs is discussed. The proposed method offers distinct advantages over eGC, especially in terms of analysis times and selectivity which can serve as a reliable platform for authenticity control of TTO.