A kind of polyphenol compound in oriental tobacco and its preparation method and application
A technology of polyphenolic compounds and oriental tobacco, applied in the field of tobacco chemistry
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[0023] Preparation method C of polyphenolic compound of the present invention 18 h 18 o 6 The preparation method comprises extract extraction, silica gel column chromatography, high-pressure liquid chromatography separation steps, specifically comprises:
[0024] A. Extraction: take oriental tobacco rhizomes, crush them to 20-40 mesh, use 80-100% ethanol to ultrasonically extract 3-5 times, each time for 30-60 minutes, combine the extracts, filter, and concentrate under reduced pressure to form an extract;
[0025]B. Silica gel column chromatography: the extract is dry-packed with 160-200 mesh silica gel column with a weight ratio of 2.5 times for silica gel column chromatography; the gradient is carried out with a chloroform-acetone solution with a volume ratio of 1:0-1:2 elute, pool identical fractions, collect eluate from each fraction and concentrate;
[0026] C. High-pressure liquid chromatography separation: the 7:3 part of the eluent in step B is further separated an...
Embodiment 1
[0035] The rhizome samples of oriental tobacco were from Baoshan, Yunnan, and the variety was Basma. Sampling 1.8 kg of oriental tobacco rhizomes was crushed to 30 mesh, extracted 3 times with 90% ethanol and ultrasonically for 35 minutes each time, the extracts were combined, filtered, concentrated under reduced pressure to obtain an extract, and 86.5 g of extract was obtained. After dissolving the extract with 2.5 times the amount of pure methanol by weight, mix the sample with 150 g of 100-mesh silica gel, pack 1.5 kg of 160-mesh silica gel into a column for silica gel column chromatography, and use a volume ratio of 1:0, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1, 1:2 chloroform-acetone gradient elution, TLC monitoring and merging the same parts to obtain 8 parts, wherein the volume ratio is 7: The chloroform-acetone eluted part of 3 was separated by Agilent 1100 semi-preparative high-performance liquid chromatography, with 48% methanol as the mobile phase, Zorbax SB-C18 (21.2×250mm, 5...
Embodiment 2
[0037] The rhizome samples of oriental tobacco were from Baoshan, Yunnan, and the variety was Basma. Sampling 1.9 kg of oriental tobacco rhizomes was crushed to 20 mesh, extracted 5 times with 99% ethanol by ultrasonic, 60 min each time, the extracts were combined, filtered, concentrated under reduced pressure to obtain an extract, and 86.5 g of extract was obtained. After dissolving the extract with 1.5 times the amount of pure methanol by weight, mix the sample with 150g of 80-mesh silica gel, pack 1.5kg of 100-mesh silica gel into a column for silica gel column chromatography, and use a volume ratio of 1:0, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1, 1:2 chloroform-acetone gradient elution, TLC monitoring and merging the same parts to obtain 8 parts, wherein the volume ratio is 7: The chloroform-acetone eluted part of 3 was separated by Agilent 1100 semi-preparative high-performance liquid chromatography, with 48% methanol as the mobile phase, Zorbax SB-C18 (21.2×250mm, 5μm) preparative...
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