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12 protocols using limonene

1

Chiral Compound Synthesis and Characterization

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All reagents and solvents were purchased from commercial sources and used as received without further purification. Melamine (C3H6N6, 99%), potassium ferricyanide (K3[Fe(CN)6], AR), (1R)-(−)-10-CSA (99%), (1S)-(−)-10-CSA (99%), L-phenylalanine (99%), D-phenylalanine (98%), L(+)-glutamic acid (99%), D-glutamic acid (98%), L(−)-carvone (99%), S-(+)-carvone (97%), +CAM, 99% and TAD, 98% were purchased from Shanghai Macklin Biochemical Co. Ltd. (China). (+)-limonene (>95%), (−)-limonene (>95%) were purchased from Tokyo Chemical Industry (TCI) Co. Ltd (Japan). NDA, 98% was purchased from ARK Pharm, Inc., USA. Ethanol (AR, ≥99.7%), concentrated sulfuric acid (AR, 95–98%), sodium chloride (NaCl, AR) and magnesium chloride hexahydrate (MgCl2·6H2O, AR) were purchased from Sinopharm Chemical Reagent Co., Ltd. (China). Sucrose (AR), Rhodamine B (RhB, AR), methyl orange (MO, 96%), alizarin yellow R (AYR, AR), L-tryptophan (99%) and D-tryptophan (98%) were purchased from Aladdin Industrial Corporation (China). Fe(phen)3Cl2 was synthesized according to previous report 28 (link).
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2

Antifungal Compounds Screening Protocol

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Miconazole, diflucan (fluconazole), and 5-fluorocytosine were purchased from Combi-Blocks Inc. (San Diego, CA, USA). Itraconazole and voriconazole were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). 2-(4-Thiazolyl) benzimidazole (thiabendazole, TBZ) was obtained from San-ai Oil Co., Ltd. (Tokyo, Japan). Amphotericin B and terpinen-4-ol were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan), and α-pinene, camphene, 3-carene, limonene, γ-terpinene, 1,8-cineole, and p-cymene were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan).
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3

Essential Oils and SARS-CoV-2 Inhibition

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All the essential oils were provided by Bio-Jourdeness International Groups Co. Ltd. (Taichung Taiwan). Citronellol, geraniol, neryl acetate, and limonene were obtained from Tokyo Chemical Industry Co., Ltd. (Tokya, Japan). Purity of these compounds were above 99%, according to the GC and 1H-NMR analysis. McCoy’s medium, fetal bovine serum (FBS), sodium pyruvate, penicillin, and streptomycin were obtained from Invitrogen (Carlsbad, CA, USA). 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and dimethylsulfoxide (DMSO) were purchased from Sigma-Aldrich. Antibody against ACE2 was obtained from Arigo Biolaboratories (Hsinchu, Taiwan). Antibody against GAPDH was obtained from Cell Signaling Technology Inc. All other chemicals were reagent grade or HPLC grade and were supplied by either Merck or Sigma-Aldrich.
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4

Characterization of Mastic Gum Compounds

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Mastic gum was purchased from Sunsho Pharmaceutical Co. Ltd. (Fujinomiya, Japan). Standard compounds, α-pinene, β-pinene, β-caryophyllene, terpineol were purchased from Sigma Aldrich Japan (Tokyo, Japan), β-myrcene, p-cymene, linallol, anethole, verbenone from Wako Pure Chemical Industries, Ltd. (Osaka, Japan), limonene and (E)-methyl isoeugenol from Tokyo Chemical Industry Co. Ltd. (Tokyo, Japan).
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5

Aroma-Loaded Emulsifier Systems Development

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Three different emulsifiers were used in this study, namely, Tween 80, soybean lecithin, and SE-15 sucrose fatty acid ester. Tween 80 was supplied by Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China), soybean lecithin was supplied by Biosharp Company (Hefei, China), and SE-15 sucrose fatty acid ester was supplied by Hangzhou Rui Lin Chemical Co. Ltd. (Hangzhou, China). Seven different aroma compounds were used in this study, namely, 3-carene, ethyl acetate, perillaldehyde, (E)-2-hexenal, carvone, α-pinene (Sigma Aldrich, Saint Louis, MO, USA), and limonene (Tokyo Chemical Industry, Tokyo, Japan). Xanthan gum (Shanghai Jingchun Biochemical Technology Co., Ltd., Shanghai, China) was used to prepare different emulsifier systems to which the thickener was added. Anhydrous ethanol (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) was added to dissolve the aroma compounds.
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6

Antcins and Terpene Compounds Bioassay

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GC376 (≥ 98%) (Cat # T5188) was purchased from TargetMol (Washington Street, Wellesley Hills, MA, USA). High glucose Dulbecco's Modified Eagle's Medium (DMEM)/high glucose containing 4 mM L-Glutamine, 4.5 g/L glucose (Cat # AE29444172) without sodium pyruvate, penicillin–streptomycin antibiotic solutions, and fetal bovine serum (FBS) were obtained from Hyclone/G.E. Healthcare Life Sciences (Logan, UT, USA). Antcins (A, B, C, H, I, and M) were isolated from fruiting bodies of T. camphoratus and A. salmonea, as described previously16 (link),55 (link), and purchased from the R&D Center of Taiwan Leader Biotechnology Corp. (Taichung, Taiwan) and purified using HPLC and FT-NMR analysis. GC-grade citronellol (Cat # C0370) and Limonene (Cat # L0132) were obtained from Tokyo Chemical Industry Co., Ltd (Toshima, Tokyo, Japan).
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7

Arecoline Delivery System Development

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The following reagents were used: arecoline hydrobromide (Sigma-Aldrich, St. Louis, Missouri, USA), Eudragit RL 100 (ERL), Eudragit RS 100 (ERS) (Rohm Gmbh, Sontheim/Brenz, Germany), polyvinylpyrrolidone K 30 (PVP), limonene, cineole, carvone, 1-octasulponic acid, and menthol (Tokyo Chemical Industry, Tokyo, Japan), polyethylene glycols 400 (PEG) (Merck Chemicals, Darmstadt, Germany). The free base of arecoline was prepared and purified by repeated extraction of aqueous arecoline hydrobromide solution (adjusted to pH 10 by 2 M NaOH) with petroleum ether [20 ]. All other chemicals and solvents were of analytical reagent grade.
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8

Evaluation of Essential Oils and Compounds

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Four types of EOs: Origanum oil (Tokyo Chemical Industry) , peppermint oil (KISHIDA chemical) , cinnamon oil (KISHIDA chemical Co., Ltd., Osaka, Japan) and clove oil (KISHIDA chemical) , and Five types of EO components: AITC (Fuji Film Wako Pure Chemical Industries, Ltd., Osaka, Japan) , carvacrol (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan) , citral (Tokyo Chemical Industry) , eugenol (Tokyo Chemical Industry) and (+) -limonene (Tokyo Chemical Industry) , were used.
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9

Monoterpene Induction in Cell Cultures

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Cells were induced with the following chemicals: Camphor (Tokyo Chemical Industry, CAT#: C0011); borneol (Tokyo Chemical Industry, CAT#: B0525); fenchone (Tokyo Chemical Industry, CAT#: F0164); fenchol (Alfa Aesar, CAT#: L03211); eucalyptol (Acros Organics, CAT#: 110340050); verbenol (Sigma Aldrich, CAT#: 247065-5G); camphene (Tokyo Chemical Industry, CAT#: C0009); carvone (Acros Organics, CAT#: 154590050); alpha-pinene (Acros Organics, CAT#: 131270050); limonene (Tokyo Chemical Industry, CAT#: L0132); linalool (Combi-Blocks, CAT#: QH-3254). All monoterpenes were flushed with argon and stored at -20°C after opening. It should be noted that the commercially available camphene used in this study is >78% pure and contains as much as 20% tricyclene.
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10

Separation and Quantification of Volatile Organic Compounds

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HayeSep â (80-100 mesh) was purchased from Supelco. Hexane for the HPLC (≥97% and redistilled), nonane, decane, undecane, tridecane, tetradecane, pentadecane, a-pinene (98%), benzothiazole (96%), b-pinene (99%), myrcene (90%), (Z)-3-hexenyl acetate (98%), ocimene [a mix of isomers (E) 70% and (Z) 30%], benzaldehyde (99%), indole (98.5%) and a-copaene (90%) were purchased from Sigma-Aldrich (Steinheim, Germany). Linalool, a-humulene (96%) and limonene (97%) were purchased from TCI-America (Portland, USA). Geranylacetone (a mixture of isomers) (96%) and (E)-b-farnesene were purchased from TCI (Tokyo, Japan). (E)-4,8-dimethyl-1,3,7 nonatriene (DMNT) and (E,E)-4,8,12-trimethyl-trideca-1,3,7,11-tetraene (TMTT) were kindly provided by Dr. Michael A. Birkett from Rothamsted Research (UK).
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