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5 protocols using 3 ethyl 2 5 dimethylpyrazine

1

Profiling Volatile Compounds in Green Coffee

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Green coffee beans (Coffea arabica, from Brazil) were bought from a commercial market in Seoul, Korea. The origin of green coffee beans was verified by the importers and experts. 2-Methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2-ethyl-3-methylpyrazine, 2-ethyl-5-methylpyrazine, 3-ethyl-2,5-dimethylpyrazine, furfural, furfuryl acetate, furfuryl propionate, furfuryl alcohol, 1-furfurylpyrrole, 4-ethylguaiacol, 2-methoxy-4-vinylphenol, 5-methylfurfural, quinoxaline, and C7–C30 alkane standard were obtained from Sigma–Aldrich Chemical Co. (St. Louis, MO, USA). 2-Ethylpyrazine, 2,3-dimethylpyrazine, 1-methylpyrrole, guaiacol, and 2-acetylfuran were bought from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS, 50-μm film thickness) solid-phase microextraction (SPME) fiber was purchased from Supelco, Inc. (Bellefonte, PA, USA).
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2

Comprehensive Protocol for Diverse Chemical Compounds

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2-Methyl-1-propanol, 3-methyl-1-butanol, 3-methyl-butanal, 3-octanone, p-xylene, styrene, benzeneacetaldehyde, phenylethyl alcohol, 4-ethyl-phenol, 2,5-dimethyl-pyrazine, trimethyl-pyrazine, and 3-ethyl-2,5-dimethyl-pyrazine were purchased from Sigma-Aldrich China Inc. (Beijing, China). The 18 amino acid standards, including glycine (Gly), asparagine (Asn), alanine (Ala), glutamic acid (Glu), lysine (Lys), arginine (Arg), proline (Pro), valine (Val), tyrosine (Tyr), isoleucine (Ile), leucine (Leu), tryptophan (Try), serine (Ser), histidine (His), threonine (Thr), methionine (Met), cysteine (Cys) and phenylalanine (Phe), were all purchased from Biosharp.
High-performance liquid chromatography (HPLC)-grade acetonitrile was supplied by Tedia Company (Fairfield, OH, USA). Phenylisothiocyanate (PITC, lot code: 10113227) was purchased from Alfa Aesar Chemical Co. Ltd (Tianjin, China), with a purity higher than 97%. Triethanolamine (TEA, analytical grade) and n-hexane (analytical grade) were purchased from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China). Pure water was obtained using a Milli-Q purification system (Millipore, Bedford, MA, USA).
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3

Flavor Compounds Analytical Protocol

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The flavor standards, methanethiol (97 %), eucalyptol (99 %), 2,5-dimethyltetrahydrofuran (98 %), 2,5-dimethylpyrazine (98 %), hexanal (98 %), 2-ethyl-5-methylpyrazine (96 %), (E)-2-octenal (97 %), 3-ethyl-2,5-dimethylpyrazine (98 %), benzaldehyde (99.5 %), (Z)-2-octen-1-ol (97 %), 2-pentylfuran (98 %), carvone (96 %), methyleugenol (98 %), anethole (99 %), (E, E)-2,4-decadienal (97 %), 2,3,5-trimethylpyrazine (99 %), (Z)-3-phenylacrylaldehyde (97 %), p-cresol (99 %), 4-hydroxy-5-methyl-3(2H)-furanone (97 %), 2,3-dimethyl-5-ethylpyrazine (99 %), dimethyl trisulfide (98 %), 2(5H)-furanone (98 %), 1-octen-3-ol (98 %), 3-furaldehyde (97 %), and 2-furanmethanol (98 %) were commercially purchased from Sigma-Aldrich (Shanghai, China). And methyl pyrazine (98 %) and benzene acetaldehyde (98 %) were purchased from TCI (Shanghai, China). Meanwhile, 2-methyl-3-heptanone (99 %) from Dr. Ehrenstorfer (Beijing, China) as the internal standard and the n-alkanes external standard (C7-C40, ≥ 97 %) from o2si Smart Solutions (Shanghai, China) were purchased.
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4

Comprehensive Coffee Compound Analysis

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The following chemicals were obtained from Sigma-Aldrich (Munich, Germany): standard caffeine (CAS: 58-08-2), trigonelline (CAS: 6138-41-6), sucrose (CAS: 57-50-1), 5-caffeoylquinic acid (5-CQA) (CAS: 906-33-2) used to represent chlorogenic acid, reference volatile compounds including; 2-furfurylthiol (CAS 98-02-2), hexanal (CAS: 66-25-1), 2-methylfuran (CAS 534-22-5), 2-butanone (CAS: 78-93-3), 2-methyl pyrazine (CAS: 109-08-0), 2,3-dimethyl pyrazine (CAS: 5910-89-4), 2-ethyl-3,5-dimethylpyrazine (CAS:13925-07-0), 2-ethyl-6-methylpyrazine (CAS: 13925-03-6), 2-ethyl-3-methylpyrazine (CAS: 15707-23-0), pyridine (CAS: 110-86-1), 2,3-butanediol (CAS: 513-85-9), benzaldehyde (CAS: 100-52-7), 4-nonanol (CAS: 5932-79-6), phenol (CAS: 108-95-2), 2,3-butanedione (CAS: 207-069-8); HPLC grade water, hexane and basic lead acetate (CAS: 301-04-2).
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5

Characterization of Volatile Compounds

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The necessary chemical standards were obtained from the suppliers shown in parenthesis. Acetic acid, 99% (Merck, Darmstadt, Germany); 3-methyl butanal, 98%; butanoic acid, 97%; pentan-2-one, 98%; dihydro-2-methyl-3(2H) furan, 97%; 2-methylpyrazine, 98%; 3-furaldehyde, 99%; 2-furamethanol, 97%; 2-acetyl furan, 99%; 2-ethylpyrazine, 98%; benzaldehyde, 98%; 5-methyl-2-furfural, 99%; 2-pentyl furan, 98%; ethyl furfural, 98%; limonene, 99%; phenyl acetaldehyde, 99%; 2-acetylpyrrole, 98%; 2-pyrrole carboxaldehyde, 97%; 3-methyl phenol, 99%; 2-ethyl-3,5-dimethylpyrazine, 98%; linalool, 98%; 5-hydroxy methyl furfural, 99%; β-cyclocitral, 97%; 2,6-dimethoxyphenol, 98%; β-caryophyllene, 98%; α-ionone, 98%; α-copaene, 99%; β-ionone, 98% (Aldrich, Steinheim, Germany).
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