Development and Characterization of a High-Purity Terpinen-4-ol Certified Reference Material by Mass Balance and qNMR

Abstract

Featured Application: This work describes the development and characterization of an SI-traceable, high-purity terpinen-4-ol (TP4O) certified reference material (CRM) to support the accurate, reliable, and comparable quantitative analysis of essential oil components across laboratories. Overall, the featured application of this work lies in strengthening the metrological traceability and analytical reliability for volatile organic compounds, with TP4O serving as a model compound for broader CRM development in applied laboratories. Terpinen-4-ol (TP4O) is a key monoterpene alcohol commonly used as a quality and authenticity marker in essential oils, cosmetics, herbal products, and pharmaceutical formulations. However, reliable and comparable quantification of TP4O across laboratories is challenged by variability in natural matrices and the limited availability of well-characterized, traceable reference materials. In this study, a high-purity certified reference material (CRM) of TP4O was developed and characterized by the National Institute of Metrology (Thailand). The material’s purity was determined using two independent and complementary approaches: a mass balance method (MB) method based on gas chromatography with flame ionization detection (GC-FID), Karl Fischer coulometric titration (KFT), and thermogravimetric analysis (TGA), and a quantitative ¹H NMR (qNMR) method employing DSS-d<inf>6</inf> as an internal standard. The purity values obtained using the MB (98.41 ± 0.09%) and qNMR (99.13 ± 0.94%) methods were statistically equivalent (p > 0.05). Based on the combined evaluation, a certified purity value of 98.77% with an expanded uncertainty of 3.05% (k = 2) was assigned. Homogeneity and short- and long-term stability assessments confirmed the suitability of the material for its intended use. This TP4O CRM provides an SI-traceable, high-purity reference to support calibration, method validation, and quality assurance in analytical applications involving essential oil components.