Thymol (Fisher Scientific, Franklin, MA, USA) and eugenol (Acros, Thermo Fisher, Franklin, MA, USA) stock solutions were prepared by pre‐dissolving phytochemicals in ethanol (Thymol and eugenol: 10 * 103 ppm for propagations, 40 * 103 ppm for EC50 assays; blend: 10 * 103 ppm Thymol + 40 * 103 ppm eugenol). Stock solutions were sterile‐filtered, aliquoted to sterile 2‐mL tubes and stored at −20 °C throughout each 6‐week experiment.
Thymol
Manufactured by Thermo Fisher Scientific
Sourced in United States, Belgium, Germany, United Kingdom
Thymol is a natural chemical compound found in various plants, particularly in the essential oils of thyme, oregano, and other herbs. It is a colorless to yellow crystalline solid with a distinctive aromatic odor. Thymol is commonly used as a preservative, antiseptic, and flavoring agent in various industries, including pharmaceuticals, cosmetics, and food processing.
Automatically generated - may contain errors
Lab products found in correlation
Methanol, by Merck Group (5 mentions)
Carvacrol, by Merck Group (5 mentions)
Eugenol, by Merck Group (4 mentions)
N hexane, by Merck Group (3 mentions)
Chloroform, by Merck Group (3 mentions)
α pinene, by Merck Group (3 mentions)
Eugenol, by Thermo Fisher Scientific (2 mentions)
Dl menthol, by Merck Group (2 mentions)
Caryophyllene oxide, by Thermo Fisher Scientific (2 mentions)
Ethanol, by Thermo Fisher Scientific (2 mentions)
Linalool, by Merck Group (2 mentions)
Geraniol, by Merck Group (2 mentions)
Camphor, by Thermo Fisher Scientific (2 mentions)
Camphene, by Merck Group (2 mentions)
Thyme oil, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Sulfuric acid, by Thermo Fisher Scientific (2 mentions)
N n dimethylformamide (dmf), by Thermo Fisher Scientific (2 mentions)
Formic acid, by Avantor (2 mentions)
Tween 80, by Merck Group (2 mentions)
26 protocols using thymol
1
Phytochemical Stock Solution Preparation
2
Maxillary Premolar Extraction Protocol
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This study was conducted at King Saud University, Saudi Arabia, from November 2018 to March 2019 after the approval of ethical committee (Ref. No. IR 0331). In this study, 60 human maxillary premolars were collected after orthodontic extractions. The teeth were cleansed and stored in 0.1% thymol solution (thymol, Fisher Scientific, NJ, USA). All teeth were mounted in orthodontic acrylic resin (Orthodontic Resin, Dentsply caulk, DE, USA) vertically, 2mm below the cemento-enamel junction (CEJ) using a polyvinyl carbonate section.
3
Fabrication of Multifunctional Nanoparticles
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Nanoparticles were prepared using an emulsification-evaporation method. The oily phase was formed by dissolving 100 mg of PLGA 50:50 lactic according to a glycolic acid ratio (Sigma-Aldrich, MA, USA) and 20 mg of thymol (MW = 150.22 g/mol; ThermoFisher, MA, USA) in 10 mL of acetonitrile (Bio-Lab Ltd., Israel) containing 0.01% Tween 80 (ThermoFisher, MA, USA). While being stirred, the aqueous phase of Solutol 0.1% (Glentham Life Sciences, UK) was added to the oily phase and stirred for 10 min using an overhead stirrer at 550 rpm. Next, the solution was transferred into a round bottom flask and connected to a rotary evaporator (Heidolph, Germany). After the solvent was entirely evaporated, the nanoparticles were centrifuged at 10,000 rpm for 5 min to remove impurities. A solution of 0.5 mg/mL thymol-loaded PLGA particles was deposited on porous titanium. mPEG-b-PLA micelles, prepared by a dialysis method as previously described [16 (link)], with a size of 30 nm, and silica nanoparticles, purchased from Nanocomposix (CA, USA), with a size of 50 nm, were deposited on porous aluminum and gold surfaces. The volume of suspended nanoparticles was 20 µL for a 1 cm by 1 cm piece of metal; evaporation was carried out overnight at room temperature.
4
Antimicrobial Treatments for S. aureus
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Approximately 1.0 × 106 CFU from exponential-phase cultures of the S. aureus chicken ready-meal isolate were subjected to the following treatments: 0.09% (wt/vol) potassium sorbate, 12% NaCl, 0.09% sodium citrate, 0.5% IGEPAL CO-630 (a nonionic, nondenaturing detergent; Merck), and 0.25% (wt/vol) thymol (Thermo Fisher Scientific)—all for 30 min at 37°C, followed by heat treatment at 50°C for 30 min. Three replicates per treatment received staining. After treatment, the cells were stained and analyzed as described above. The effects of these treatments on cell viability were also recorded using plate counting.
5
Antimicrobial Effects of Organic Compounds
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Citric acid, sorbic acid, benzoic acid, butyric acid, hexanoic acid, thymol, vanillin, carvacrol, and eugenol were purchased from Alfa Aesar (Thermo Fisher GmbH, Kandel, Germany). Stock solutions of organic acids and NIC were prepared in BHI and BHI supplemented with 70% (v/v) ethanol, respectively. All solutions were buffered to pH 6.5 and filter-sterilized, then they were conserved at +4 °C and brought back to room temperature before each use.
6
Antibiotic Resistance Profiling of MRSA and CRPA
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Each MRSA and CRPA strains used in this study were obtained from a different patient with post-tympanostomy tube otorrhea at Chosun University Hospital. Clinical isolates were subcultured for 24 h in blood agar plates (Hanil-KOMED, Seongnam, Korea). They were inoculated into the GP and GN cards of VITEK 2 system (bioMérieux Inc., Durham, NC, USA), and then tested for the identification of S. aureus and P. aeruginosa according to the recommendations of the manufacturer. The MRSA and CRPA strains were phenotypically characterized by antibiotic susceptibility test cards (AST-P601 for MRSA, AST-N225 for CRPA) of VITEK 2 system (bioMérieux Inc.). The MRSA and CRPA isolates were cultured at 37 °C on Mueller–Hinton agar (Difco Laboratories, Sparks, MD, USA) plates. Thymol was purchased from ThermoFisher Scientific (Waltham, MA, USA), a stock solution (250 mg/mL) of which was prepared by dilution in ethanol (1.25%, Merck KGaA, Darmstadt, Germany).
7
Characterization of Terpene Compounds
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The chemicals used were the following: DL-menthol (CAS [89-78-1], ≥95%) and DL-camphor (CAS [76-22-2], ≥96%)) from Sigma-Aldrich (St. Louis, MO, USA), thymol (CAS [89-83-8], ≥98%) and L-borneol (CAS [464-45-9], ≥97%)) from Alfa Aesar (Haverhill, MA, USA). Deuterated dimethyl sulfoxide (DMSO-d6, CAS [67-68-5]) and chloroform (CDCl3, CAS [865-49-6]) and tetramethylsilane (TMS, CAS [75-76-3]) used in NMR experiments was purchased from Sigma-Aldrich (St. Louis, MO, USA).
8
Analytical Profiling of Volatile Constituents
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All the chemicals including methanol, chloroform, and n-hexane were of analytical grade and purchased from Sigma–Aldrich, Hamburg, Germany. Pure volatile constituents or enriched fractions of volatile constituents, such as thymol, δ3-carene, α-pinene (Alfa Aesar, Lancashire, UK), n-hexadecanoic acid, caryophyllene oxide, (Z)-9-Octadecenoic acid methyl ester, and 8,11-Octadecadienoic acid, methyl ester (enriched fractions), were available and used for co-injection/comparative analysis.
9
Evaluating Insecticidal Potential of Essential Oil Constituents
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High purity essential oil components carvacrol, geraniol, eugenol, methyl eugenol, trans-cinnamaldehyde, citronellic acid, (±)-citronellal, α-pinene, linalool, R (+)-limonene, eucalyptol, (−)-terpinen-4-ol, and menthone were obtained from Sigma-Aldrich (St. Louis, MO), whereas thymol and (±)-camphor were obtained from Alfa Aesar (Hill, MA) (Table S1 ). These active constituents are found in various aromatic plants (Table S1 ). All fifteen essential oil components (Table S1 ) were selected based upon the previous toxicity literature on different urban and agricultural pests22 (link)–31 (link). The positive controls dichlorvos (≤100% purity) and bifenthrin (98% purity) were obtained from Sigma Aldrich and Chem Service Inc. (West Chester, PA), respectively. Analytical grade solvents such as acetone, ethanol and dimethyl sulfoxide (DMSO) were purchased from Fisher Scientific (Pittsburgh, PA). Buffer salts and other reagents used for preparation of HEPES (4-(2-hydroxyethyl)−1-piperazineethanesulfonic acid)-buffered physiological saline were purchased from Sigma-Aldrich, Fisher Scientific and Avantor Performance Materials, LLC (Center Valley, PA).
10
Volatile Constituents Analysis Protocol
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All the chemicals including methanol, chloroform, and n-hexane were of analytical grade and purchased from Sigma–Aldrich, Germany. Pure volatile constituents or enriched fractions of volatile constituents such as camphene (Sigma–Aldrich, Burlington, MA, USA), heptacosane, carvacrol (Sigma Aldrich, Shanghai, China), thymol (Alfa Aesar, Lancashire, UK), piperitone, caryophyllene oxide, and spathulenol (enriched fractions) were available and used for co-injection/comparative analysis.
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