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3 protocols using terpinene 4 ol

1

Quantifying Antioxidant Potential of Terpenes

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The terpenes (−)-isopulegol (≥99%), menthol (≥99%), p-cymene (≥99%), eucalyptol (≥99%), (R)-(+)-pulegone (97%), γ-terpinene (97%), α-terpinene (≥95%), linalool (≥97%), (S)-(+)-carvone (≥96%), citronellal (≥95%), (−)-terpinene-4-ol (≥95%), citral (≥95%), menthone (≥90%), farnesene (mixtures of isomers, ≥90%), α-phellandrene (≥90%), β-myrcene (≥90%), and TPTZ (2,4,6-Tri(2-pyridyl)-s-triazine)) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Acetic acid (ACS), FeCl3 × 6H2O, FeSO4 × 7H2O, phosphoric acid (ACS), hydrochloric acid (ACS), chloroform (ACS), ethyl acetate (ACS), 2-butanone (ACS), and methanol (analytic purity grade) were obtained from Polish Reagents (Gliwice, Poland). The standards trolox ((±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid, 97%) and gallic acid (>98%) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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2

Characterization of Essential Oils

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The essential oils (EOs) used in the tests were purchased from “JSC Naujoji Barmune”, a retailer of EOs in Lithuania. The following EOs were used: thyme (Thymus vulgaris), eucalyptus (Eucalyptus globulus), spearmint (Mentha spicata), lavandin (Lavandula × hybrida), East-Indian lemongrass (Cymbopogon flexuosus), and clove (Eugenia caryophyllus). All EOs were labelled with the common and scientific names of the plant species they were extracted from, as well as with series numbers and expiry dates. However, the origin of the plants used to extract the EOs was not indicated.
Analytical standards used in the experiment were purchased from different companies: (R)-(−)-carvone from Toronto Research Chemicals Inc. (Toronto, ON, Canada); limonene (sum of enantiomers, analytical standard, ≥98% (GC)), myrcene (analytical standard, ≥95% (GC)), and γ-terpinene (analytical standard, ≥97% (GC)) from Fluka (Charlotte, NC, USA); cis-sabinene hydrate (analytical standard, ≥97% (GC)) and terpinene-4-ol (sum of enantiomers, analytical standard, ≥95% (GC)) from Sigma-Aldrich (Darmstadt, Germany).
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3

Hydrodistillation and GC-MS Analysis of Essential Oil

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Analytical grade n-heptane, n-hexane, methanol, ethyl acetate, and acetonitrile were purchased from Avantor Performance Materials, (Gliwice, Poland), while tert-butyl methyl ether (MTBE) was from Acros Organics (part of ThermoFisher Scientific, Belgium). 99.999% pure helium 5.0 was used for GC/MS analysis (PGNiG, Poland). A Millipore purification system (France) was used to generate HPLC-grade water. Standard samples of p-cymene, eucalyptol, 1octen-3-ol, camphor, terpinene-4-ol, linalyl acetate, thymol, borneol, carvacrol, carvone, linalool, amoxicillin, streptomycin and caryophyllene oxide were obtained from Sigma-Aldrich (Germany).
Cell culture-grade DMSO was purchased from AppliChem Panreac. LB Broth (Miller; molecular biology grade), PBS (phosphate-buffered saline) and MacConkey agar were purchased from Fisher Scientific UK Ltd. Identification was performed by specialists in botany, Mrs Krystyna Dąbrowska from The Botanical Garden of Marie Curie University. After drying, the plant was homogenised and a sample (50 g) was hydrodistilled with 400 mL of distilled water for 3 h in a Deryng apparatus. The EO was stored over anhydrous sodium sulfate at 4°C until analysis.
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