Determination of Α- and Β-Thujone in Wormwood and Sage Infusions of Greek Flora and Estimation of their Average Toxicity

Abstract

Artemisia absinthium (wormwood) and Salvia sp. (sage) are used as additives to pharmaceuticals, food and alcoholic beverages while in Greece both herbs are also consumed as infusions. The compounds α- and β-thujone, are monoterpenes, which are found in various preparations of these herbs. According to organizations (EMA, SCF) and clinical studies, α- and β-thujone have shown neurotoxic effects in humans over established limits. In this study wormwoodandsageinfusions were investigated in terms of the relative % content of the volatile constituents and the concentrations of α- and β-thujone by gas chromatography-mass spectrometry (GC-MS). In sage infusions, α-thujone was the predominant isomer while in wormwood β-thujone was dominated. Our results revealed that most of the above herbal infusions exceed thujone limits as set by EMA, which are 3 mg/day for wormwood and 5 mg/day for sage. Furthermore, the average toxicity as performed by the Microtox® Analyzer showed higher levels of toxicity in sage infusions compared to the wormwood. No correlation was found between thujone content and toxicity levels of both sage and wormwood infusions.

Development of a PCR High‐Resolution Melt Assay for Artemisia absinthium (Wormwood) and a Triplex Assay with Two Additional “Unregulated Legal High” Species Datura stramonium (Jimson Weed) and Merremia tuberosa (Hawaiian Woodrose),

“…A. absinthium contains thujone, and when consumed in large doses, along with large quantities of alcohol, can induce convulsions. GC‐MS is a widely used method to detect and differentiate drug analytes and has been used to identify and quantitate α‐ and β‐thujone . However, unlike DNA typing methods, GC‐MS cannot determine the source of the thujone, sage ( Salvia officinalis ), or wormwood .…”

“…Individuals who frequently ingested absinthe would experience epileptic‐like episodes. Originally thought to act on the same receptors as tetrahydrocannabinol (THC), researchers later realized that the two components of wormwood are α‐thujone and more prominently, β‐thujone, which block the gamma‐aminobutyric acid (GABA) receptors in the brain, causing convulsions in high doses . Researchers have shown that thujone alone was not responsible in this absinthism effect and that higher levels of alcohol at the time might have increased the effects of thujone in an individual .…”

“…Thujone can be detected and quantitated using gas chromatography–mass spectrometry (GC‐MS) . The U.S. Food and Drug Administration requires that absinthe has to be “thujone‐free” and contain less than 10 ppm in food and drinks; however, the compound is not covered by the Drug Enforcement Act, allowing individuals to legally purchase and consume wormwood to obtain an “unregulated legal high” .…”

“…Preparation of these “unregulated legal high” substances for recreational use can occur in common spaces, such as food preparation areas, and there is a safety concern of cross‐contamination where other individuals can be exposed unknowingly, including children. Researchers have determined that the thujone levels are over the toxicity limit in wormwood and sage infusions . Interestingly, α‐thujone and β‐thujone were found in higher quantities in sage than wormwood .…”

“…Researchers have determined that the thujone levels are over the toxicity limit in wormwood and sage infusions . Interestingly, α‐thujone and β‐thujone were found in higher quantities in sage than wormwood . Wormwood also has been characterized microscopically using a scanning electron microscope (SEM) and various cell staining techniques to identify chemical structures of the plants’ seeds .…”