A method for separating cyanidin in Nitraria by high-speed countercurrent chromatography
A technology of high-speed countercurrent chromatography and cyanidin, which is applied in the field of high-speed countercurrent chromatography to separate cyanidin-3-O--diglucoside from Nitraria japonica, achieving the effects of good repeatability, convenient operation and simple process
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Embodiment 1
[0023] 1. Preparation of crude extract
[0024] Add 100mL methanol (2% formic acid v / v) to 100g dried fruit of Nitraria tangutica and extract for 24h, repeating 3 times. After 3 times of extracts were combined, filter paper was used to remove insoluble matter, protein, and polysaccharides. After the filtrate was collected, the methanol was concentrated by a rotary evaporator, and the extract was redissolved in distilled water (2% formic acid) and applied to D-101 macroporous resin (50mm×800mm). The macroporous resin was pretreated with methanol and water. After loading the sample, flush the column with 3 times the column volume of distilled water to remove sugars, amino acids and other impurities. Then the column was flushed with methanol, and the effluent was collected, concentrated under reduced pressure, and dried.
[0025] 2. Preparation of cyanidin-3-O-(trans-p-coumarin)-diglucoside by high-speed countercurrent separation
Embodiment 2
[0037] 1. Preparation of crude extract
[0038] 100g of dried Nitraria tangutica fruit was added to 100mL of methanol (2% formic acid) to extract for 24h, repeated 3 times. After 3 times of extracts were combined, filter paper was used to remove insoluble matter, protein, and polysaccharides. After the filtrate was collected, the rotary evaporator was concentrated to remove methanol, and the extract was reconstituted with distilled water (2.% formic acid) and applied to D-101 macroporous resin (50mm×800mm). The macroporous resin was pretreated with methanol and water. After loading the sample, flush the column with 3 times the column volume of distilled water to remove sugars, amino acids and other impurities. Then the column was flushed with methanol, and the effluent was collected, concentrated under reduced pressure, and dried.
[0039] 2. High-speed countercurrent separation
[0040] Prepare a two-phase solvent system of methyl tert-butyl ether: n-butanol: acetonitrile...
Embodiment 3
[0043] 1. Preparation of crude extract
[0044]100g of dried Nitraria tangutica fruit was added to 100mL of methanol (2% formic acid) to extract for 24h, repeated 3 times. After 3 times of extracts were combined, filter paper was used to remove insoluble matter, protein, and polysaccharides. After the filtrate was collected, the methanol was concentrated by a rotary evaporator, and the extract was redissolved in distilled water (2% formic acid) and applied to D-101 macroporous resin (50mm×800mm). The macroporous resin was pretreated with methanol and water. After loading the sample, flush the column with 3 times the column volume of distilled water to remove sugars, amino acids and other impurities. Then the column was flushed with methanol, and the effluent was collected, concentrated under reduced pressure, and dried.
[0045] 2. High-speed countercurrent separation
[0046] Prepare a two-phase solvent system of methyl tert-butyl ether: n-butanol: acetonitrile: water (0....
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