A method for extracting Anthocyanin Naturals in mulberries. Take fresh mulberry fruits, remove the stems, beat into a homogenate with a mixer, take 10g of the homogenate and use the deep eutectic solvent choline chloride-lactic acid (molar ratio 1:3, Water content 0%wt) is the extraction solvent, extract for 10 minutes under the conditions of solid-liquid ratio 1g: 30mL, ultrasonic power 100W, electrostatic field strength 4kV, extraction temperature 50°C to obtain anthocyanin extract; add to the above anthocyanin extract 5g of attapulgite-loaded chitosan cross-linked cyclodextrin complex was shaken on a shaking table at 30°C and 100r/min for 4 hours, and the precipitate was collected after centrifugation, washed with deionized water ultrasonically for 3 times, and centrifuged to get the adsorbed on the attapulgite load. Anthocyanin precipitation on chitosan cross-linked cyclodextrin complex; add ethanol with a mass fraction of 80% to Anthocyanin Naturals precipitation for ultrasonic desorption for 1.0 h, centrifuge to obtain anthocyanin supernatant containing ethanol, and put anthocyanin on The supernatant is filtered through a 0.22um microporous membrane to obtain a purified Anthocyanin Naturals solution containing ethanol. At 50±10°C, concentrate under reduced pressure to remove the ethanol in the anthocyanin solution, and remove the ethanol in the anthocyanin solution by freeze-drying in vacuum. For excess moisture, get Anthocyanin Naturals. The extraction rate of anthocyanins was 9.2427 mg/g.
A method for isolating and preparing Anthocyanin Naturals monomers from Lycium barbarum fruit, comprising the following steps:
⑴Raw material pretreatment: select and remove impurities from the black fruit wolfberry fruit, rinse with water, wash away the surface soil, and obtain the processed black fruit wolfberry fruit;
(2) Prepare a methanol solution with a volume concentration of 2% formic acid: mix 200 mL of formic acid and 9800 mL of methanol evenly;
(3) The methanol solution of formic acid obtained in the step (2) is added to the fruit of Lycium ruthenicum fruit, and extracted at room temperature at a ratio of 1 g: 1 ~ 5 mL in a dark state at room temperature, and the number of extractions is 1 ~ 3 times. 12-24 h each time, combined to obtain the extract;
(4) Use filter paper to remove insolubles, proteins, and polysaccharides from the extract to obtain a filtrate, which is vacuum evaporated and concentrated to a paste on a rotary evaporator at 40-50°C to obtain the Anthocyanin Naturals extract of Lycium barbarum, and recover Methanol;
(5) Treatment of macroporous adsorption resin: Soak D-101 macroporous adsorption resin in methanol with a volume concentration of 95% for 24 hours, then wash with methanol with a volume concentration of 95%, and then wash with deionized water; The porous adsorption resin is slowly loaded into the resin column, and then rinsed with deionized water until the particles in the column are clean and no longer settles, that is, the processed macroporous adsorption resin is obtained;
(6) the macroporous adsorption resin after the above-mentioned treatment after redissolving the anthocyanin extract of Lycium barbarum with a volume concentration of 2%; after loading the sample, flush the column with distilled water of 4 to 6 times the column volume, and then Then flush the column with 3~6 column volumes of methanol to desorb the Anthocyanin Naturals compounds and collect the effluent;
(7) The effluent is concentrated under reduced pressure to obtain crude anthocyanins, and methanol is reclaimed simultaneously;
(8) Inject the Anthocyanin Naturals crude product into a high-efficiency semi-preparative chromatographic column under the conditions of a column temperature of 25-40°C, a flow rate of 15-25 mL/min, an injection volume of 200-600 mg, and a detection wavelength of 525nm. Semi-preparative chromatographic separation, according to the peak time to collect monomer petunia-3,5-O-diglucosides with lower purity, petunia-3-O-rutinoside (trans-p-coumaroyl)-5-O -glucosides; then use the high performance liquid chromatography system to detect under the conditions of column temperature of 25~40℃, flow rate of 0.6~1.0 mL/min, injection volume of 10 ul, and detection wavelength of 525nm. When the sample purity is lower than At 90%, the high-efficiency semi-preparative chromatographic column is used again for secondary separation to obtain anthocyanin monomers with a purity ≥ 90%; finally, the Anthocyanin Naturals monomers are sequentially concentrated under reduced pressure on a rotary evaporator and vacuum freeze-dried To constant weight, the finished product of Anthocyanin Naturals monomer is obtained.