Hexadecanoic acid

All the chemicals and reagents involved in the extraction processes, total polyphenolic content, antioxidant activity experiments and the chemical standards (1-hexanol, 2-Thujene, 3-octanone, 5-Amino-1-ethylprazole, Allylbenzene, Camphene, Caryophyllene oxide, Decanoic acid, Docosane, Germacrene D, Hexadecane, HexaDecanoic acid, Lavandulol, Limonene, Linalool, Linalool acetate, Nerol, Nerolidol, Ninanal, TetraDecanoic acid, Tricosane, α-Copaene, α-Humulene, α-Ter-pinene, β-Bourbonene, β-Caryophyllene, β-Copaene, β-Cubenene, β-Farnesene, β-Gurjunene, β-Myrcene, β-Pinene) used in the identification of phenolic and aromatic compounds from the extracts were acquired from Sigma Aldrich Company (St. Louis, MO, USA).

To validate the results obtained using GC-TOF-MS, a bioassay was performed using commercial palmitic acid (hexadecanoic acid) (Sigma Aldrich) and a uredospore solution of A. psidii MF-1 (10⁵ per Petri dish) diluted in mineral oil (Sigma Aldrich). The palmitic acid was diluted in dichloromethane (JT Baker) at concentrations of 0 (control), 0.5, 2.5, 5.0, 10 and 20 ppm and combined with the solution of uredospores before inoculation in Petri dishes with solid water agar medium (8 g L^-1) and incubation for 24 h at 20°C in the dark. Germination rates were obtained as described above. The experiment was conducted in a randomized block design with six hexadecanoic acid concentrations using three replicates (Petri dishes) per treatment.

Acetonitrile, chloroform, ethyl acetate and methanol were purchased from J. T. Baker (München, Germany), and D-chloroform, formic acid 98%, Casein-Peptone Soymeal-Peptone (CASO), Potato Dextrose (PD), Luria–Bertani broth (LB), sodium chloride (NaCl), potassium chloride (KCl), potassium dihydrogen phosphate (KH₂PO₄), D-methanol and trifluoroacetic acid (TFA) were purchased from Carl Roth GmbH (Karlsruhe, Germany). Bacto malt extract, Bacto peptone and agar were from BD (La Point de Claix, France), and D-glucose was from Merck (Darmstadt, Germany). Disodium hydrogen phosphate (Na₂HPO₄) was purchased from J. T. Baker (The Netherlands), crystal violet from Fluka (Steinheim, Germany) and tetracycline and Potato Dextrose agar (PDA) from Sigma Aldrich (Taufkirchen, Germany). The fatty acids hexadecanoic acid (palmitic acid), (9E)-9-hexadecenoic acid (palmitelaidic acid), (9Z)-9-hexadecenoic acid (palmitoleic acid), (9Z)-9-Octadecenoic acid (oleic acid), (9Z,12Z)-octadeca-9,12-dienoic acid (linoleic acid), (6Z,9Z,12Z)-octadeca-6,9,12-trienoic acid (γ-linolenic acid) and (5Z,8Z,11Z,14Z)-Eicosa-5,8,11,14-tetraenoic acid (arachidonoic acid) were purchased from Sigma-Aldrich, St. Louis, MO, USA, while (7Z,10Z)-hexadeca-7,10-dienoic acid came from Larodan Inc., Monroe, LA, USA.

The dichloromethane (p.a., ≥99% purity) and ethanol (p.a., ≥99% purity) were supplied by Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA). The pyridine (p.a., ≥99.5% purity), N,O-bis(trimethylsilyl)trifluroacetamide (99% purity), trimethylchlorosilane (99% purity), tetracosane (99% purity), hexadecanoic acid (≥99% purity), pentadecan-1-ol (99% purity), stigmasterol (95% purity), and ursolic acid (≥98% purity) were supplied by Sigma Chemical Co (Madrid, Spain). The gallic acid (≥97.5% purity) was purchased from Sigma-Aldrich (Merck, Darmstadt, Germany). The Dulbecco’s Modified Eagle’s Medium (DMEM), sodium bicarbonate, sodium pyruvate, non-essential amino acids, L-glutamine, glucose, phenol red, trypsin-ethylenediamine tetraacetic acid (EDTA) solution 1X and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The fetal bovine serum (FBS), and penicillin-streptomycin were obtained from Gibco (Carlsbad, CA, USA).

The synthetic standards including (1R)-(+)-α-pinene, (-)-β-pinene, β-myrcene, (S)-(-)-limonene, linalool, octanal, nonanal, decanal, benzaldehyde, butylated hydroxytoluene, methyl benzoate, (-)-bornyl acetate, 6-methyl-5-hepten-2-one, geranyl acetone, decanoic acid, undecanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, myristoleic acid, palmitoleic acid, linoleic acid, oleic acid, (+)-longifolene, caryophyllene oxide, (E)–β-caryophyllene, undecane, dodecane, pentadecane, hexadecane and quercetin were obtained from Sigma-Aldrich (>98% purity).

Aflatoxins B₁, B_2, G₁ and G₂ were purchased from Supelco (Bellefonte, Pennsylvania, USA), apigenin (≥ 99%), luteolin (≥ 97%), rutin hydrate (≥ 94%) quercetin dehydrate (≥ 98%), octadecanoic acid (≥ 98.5%), glutamic acid, myristoleic acid, tetradecanoic acid, hexadecanoic acid, (Z)-9-hexadecenoic acid, linoleic acid, (Z)-9-octadecenoic acid, octadecanoic acid, (Z, Z)-9,12-octadecadienoic acid, oleic acid, eicosanoic acid and amino acid standard solution (AAS 18) were purchased from Sigma-Aldrich (Chemie GmbH, Munich, Germany).

Dichloromethane (p.a., ≥99% purity) was purchased from Fisher Scientific (Thermo Fisher Scientific, Waltham, MS, USA). Anhydrous pyridine (99.8% purity), N,O-bis(trimethylsilyl)-trifluoroacetamide (99% purity), trimethylchlorosilane (99% purity), tetracosane (≥99% purity), pentadecan-1-ol (99% purity), hexadecanoic acid (≥99% purity), β-stigmasterol (95% purity) and vanillin (99% purity), were supplied by Sigma Chemical^© (Madrid). Dimethyl sulfoxide (DMSO) was supplied from Sigma Aldrich^© (Darmstadt, Germany), BHIB and Mueller-Hinton mediums were provided by BIOKAR^© (BIOKAR Diagnostics, Allonne, France).