GC-MS analysis of volatile compounds from pine tar using DLLME and cytotoxicity assessment

Merve Nesetoglu; Selen Al; Gulsum Altiparmak-Ulbegi; Aykut Kul; Olcay Sagirli

doi:10.62063/ecb-65

Authors

Merve Nesetoglu Istanbul University https://orcid.org/0009-0005-2367-5793
Selen Al Istanbul University https://orcid.org/0009-0009-2014-1998
Gulsum Altiparmak-Ulbegi Istanbul University https://orcid.org/0000-0002-2585-1931
Aykut Kul Istanbul University, https://orcid.org/0000-0002-1195-0939
Olcay Sagirli Istanbul University https://orcid.org/0000-0002-0434-2150

DOI:

https://doi.org/10.62063/ecb-65

Keywords:

DLLME, GC–MS, Pine tar

Abstract

Pine tar has long been valued for its dermatological and antimicrobial properties; however, its volatile and semi-volatile component profile remains underexplored from a sustainable bioprospecting perspective. In this study, we combined dispersive liquid–liquid microextraction (DLLME) with GC-MS to develop a rapid, green workflow for profiling pine-tar volatiles, achieving greater than or equal to 50-fold enrichment from a 100 mg sample in under 10 min. GC-MS analysis on a TRB-5MS column (90 min gradient) resolved 45 compounds (match ≥ 77 %), with sesquiterpenes accounting for 71.3 % of the total area (isolongifolene 30.3 %; isobornyl acetate 16.0 %; borneol 12.4 %; (+)-longicyclene 12.5 %). Monoterpene alcohols comprised 4.4 %, while minor oxidized derivatives contributed < 3 %.
Compared to conventional hydrodistillation, DLLME reduced chlorinated solvent usage by 90 % and enhanced recovery of high-boiling terpenoids by ~ 25 %. Furthermore, the cytotoxic effect of pine tar was investigated on human umbilical vein/vascular endothelium cells (HUVEC) using 3-[4,5-dimethylthiazol-2-yl]-5[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium (MTS) cell viability assay, revealing a low level of toxicity. The resulting chemical fingerprint high lights the biotechnological potential of sesquiterpene scaffolds such as isolongifolene for microbial production and biocatalytic transformations. It also supports enzyme-based functionalization strategies for isobornyl acetate and borneol, and opens avenues in sustainable perfumery, pharmaceutical intermediates, and biofuel applications. This semi-quantitative and eco-friendly platform offers a practical foundation for the biotechnological valorization and green production of pine-tar terpenoids. In addition, the observed low cytotoxicity of pine tar contributes to the preliminary safety assessment of this natural product on human endothelial cells.

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