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ARS Home » Southeast Area » Oxford, Mississippi » Natural Products Utilization Research » Research » Publications at this Location » Publication #394465
Research Project: Biobased Pesticide Discovery and Product Optimization and Enhancement from Medicinal and Aromatic Crops

Location: Natural Products Utilization Research

Title: Development of a GC/Q-ToF method coupled with headspace solid-phase microextraction to evaluate the in vitro metabolism of ß-Caryophyllene

Author
item LEE, JOSEPH - University Of Mississippi
item Wang, Mei
item MONDAL, GOUTAM - University Of Mississippi
item KHAN, IKHLAS - University Of Mississippi
item YATES, CHARLES - University Of Mississippi

Submitted to: Molecules
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/26/2022
Publication Date: 11/2/2022
Citation: Lee, J., Wang, M., Mondal, G., Khan, I.A., Yates, C.R. 2022. Development of a GC/Q-ToF method coupled with headspace solid-phase microextraction to evaluate the in vitro metabolism of ß-Caryophyllene. Molecules. https://doi.org/10.3390/molecules27217441.
DOI: https://doi.org/10.3390/molecules27217441

Interpretive Summary: Although many technological advances have been made in the field of analytical chemistry, sample preparation remains both a challenging and time-consuming process. Many techniques often require that the analyte be extracted from any sample matrix in order to be analyzed by instrumentation. This does not only increase the sample preparation time, but also introduces the possibility of errors entering into the analytical process. In the present study, a GC/Q-ToF coupled with headspace solid-phase microextraction (HS-SPME) method was developed and validated to evaluate the In vitro metabolism of ß-caryophyllene from Copaiba essential oil. The developed method eliminated the need to remove buffers and salts commonly present in biological matrices. No solvent and minimal sample preparation is required. This method could be adapted to evaluate numerous volatiles and/or semi-volatiles in complex natural products matrices.

Technical Abstract: Sample preparation remains both a challenging and time-consuming process in the field of bioanalytical chemistry. Many traditional techniques often require multi-step processes, which can introduce additional errors to the analytical method. Given the complexity of many biological matrices, thorough analyte extraction presents a major challenge to researchers. In the present study, a headspace solid-phase microextraction (HS-SPME), coupled with GC/Q-ToF, method was developed to quantify in vitro metabolism of ß-caryophyllene by both human liver microsome (HLM) and S9 liver fractions. Validation of the method was demonstrated both in terms of linearity (R2 = 0.9948) and sensitivity with a limit of detection (LOD) of 3 ng/mL and a limit of quantitation (LOQ) of 10 ng/mL. In addition, the method also demonstrated both inter and intra-day precision with the relative standard deviation (RSD) being less than 10% with four concentrations ranging from 50-500 ng/mL. Since this method requires no solvents and minimal sample preparation, it provides a rapid and economical alternative to traditional extraction techniques. The method also eliminates the need to remove salts or buffers, which are commonly present in biological matrices. Although this method was developed to quantify in vitro metabolism of one analyte, it could easily be adapted to detect or quantify numerous volatiles and/or semi-volatiles found in biological matrices.