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4 protocols using ethyl isovalerate

1

Comprehensive Commercial Standards for Analysis

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A total of 66 commercial standards (analytical reagent grade, ≥97% purity) were used. Ethyl acetate, acetic acid, Ethyl lactate, 1-propanol, acetal, acetaldehyde, 3-methyl butanol, furfural, ethyl palmitate, isobutanol, methanol, 3-methylbutyraldehyde, ethyl butyrate, ethyl propionate, meso-2,3-butanediol, butyrate, 1-butanol, (2R,3R)-(-)-2,3-butanediol, 2-butanol, hexanoic acid, ethyl hexanoate, 3-hydroxy-2-butanone, ethyl isovalerate, furfuryl alcohol, isobutyric acid, hexyl hexanoate, ethyl valerate, 1-hexanol, propionic acid, acetone, ethyl laurate, 2-methylbutyraldehyde, isoamyl acetate, 2-butanone, ethyl isobutyrate, ethyl phenylacetate, nonanoic acid, propyl acetate, isobutyraldehyde, ethyl octanoate, 1,1-diethoxy-2-methylbutane, ethyl linoleate, valeric acid, ethyl oleate, ethyl heptanoate, ethyl formate, phenylethyl alcohol, ethyl hexadecenoate, 1-pentanol, ethyl tetradecanoate, diethyloxymethane, 2-pentanone, 2-methyl butanol, isovaleric acid, ethyl nonanoate, 2-pentanol, 1,1-diethoxy-3-methylbutane, benzaldehyde, ethyl decanoate, heptanoic acid, octanoic acid, 2-ethoxy-5-methylfuran, 1-propanal, butyl hexanoate, ethyl phenylpropionate, and benzyl alcohol were purchased from Sigma-Aldrich (Shanghai, China).
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2

Cultivation and Flavor Analysis of Aspergillus oryzae

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Soybeans, raw wheat flour and edible salt (NaCl) were purchased from a local supermarket (Zhenjiang, China, 2021). 3-Methylbutanal, ethanol, 3-methyl-1-butanol, ethyl propanoate, ethyl acetate, ethyl isovalerate, 2-phenylethanol, benzeneacetaldehyde, 3-(methylthio) propyl acetate, ethyl 3-(methylthio) propionate, methyl benzoate, (E)-β-damascenone, 2-phenylethyl acetate, 2-furanmethanol, 3-methylthio-1-propanol, 3-methylthio propanal, 2,6-dimethyl pyrazine, trimethyl pyrazine, 2-acetyl pyrroline, 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) and 4-hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF) were all ordered from Sigma-Aldrich (Shanghai, China, 2021). 3-Furaldehyde, 4-ethylphenol, 4-ethylguaiacol, 4-vinylguaiacol, 3-methylbutanoic acid, acetic acid, 2-methylbutanoic acid, dimethyl trisulfide, 1-octen-3-ol and C6-C33 n-alkanes were provided by Aladdin Holdings Group (Shanghai, China, 2021). Other analytically pure reagents were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China, 2021). A. oryzae 3.042 were obtained from the Guangdong Institute of Microbiology (Guangzhou, China, 2021).
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3

Aroma Compound Characterization Protocol

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1,4-Cineole (98%), 1-decanol (99.9%), 1-hexanol (99.9%), 1-octanol (≥99%), 2-heptanone (99%), 2-nonanone (97%), 2-undecanone (97%), 3-hexen-1-ol (98%), 5-methyl-2-furfural, benzaldehyde (≥99.5%), camphor (99%), decanal (95%), diethyl succinate (≥99%), ethyl 2-methylbutanoate (99%), ethyl butanoate (99%), ethyl decanoate (≥98%), ethyl heptanoate (≥98%), ethyl hexanoate (99%), ethyl isobutanoate (≥98%), ethyl isovalerate (98%), ethyl nonanoate (≥98%), ethyl octanoate (99%), eucalyptol (99%), furfural (99%), hexyl acetate (98%), isoamyl acetate (≥99%), isoamyl alcohol (98%), limonene (99%), linalool oxide (97%), nonanal (≥95%), octanal (≥98%), phenylacetaldehyde (90%), phenylethyl acetate (99%), phenylethyl alcohol (99%), tartaric acid (≥99.5%), α-pinene (99%)), β-cyclocitral (90%), β-damascenone (≥98%), α-ionone (85%), and β-linalool (80%) were supplied by Sigma-Aldrich (Madrid, Spain). Ethanol (99.9%) was purchased from ERBA Reagents (Val de Reuil, France).
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4

Volatile Compound Extraction and Analysis

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Furfural, ethyl isovalerate, hexanol, isoamyl acetate, γ-butyrolactone, benzaldehyde, hexanoic acid, ethyl hexanoate, limonene, ethyl 2-furoate, 1-octanol, 2-nonanone, 2-nonanol, 2-phenylethanol, diethyl succinate, ethyl octanoate, octanoic acid, β-cyclocitral, 2-phenylethyl acetate, nonanoic acid, ethyl nonanoate, decanoic acid, β-damascenone, ethyl decanoate, tetradecane, isoamyl octanoate, lilial, dodecanoic acid, ethyl dodecanoate, hexadecane, tetradecanoic acid, ethyl pentadecanoate, methyl pentadecanoate and ethyl hexadecanoate were purchased from Sigma-Aldrich-Fluka (St. Louis, Missouri, USA). Aliphatic hydrocarbons (retention index mixture for GC (KI) with purity above 95%) were also supplied by Sigma-Aldrich.
A 2 cm long Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS) fibre was supplied by Supelco (Bellefonte, PA, USA). Before extraction the fibre was conditioned according to the manufacturer's recommendations.
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