( 1) Utilize Yinghong No. 9 black tea as the raw product, pass through a 60-mesh filter after squashing, include 50% ethanol solution according to the solid-to-liquid ratio (w/v) of 1:10, reflux removal for 3 times, combine the essences as well as revolve to concentrate to ethanol-free;
( 2) Utilize ethyl acetate to draw out the concentrated extract of black tea, the quantity ratio of the focused essence of black tea to ethyl acetate is 1:1, the number of removals is 3 times, the ethyl acetate essence is incorporated as well as turned to concentrate to eliminate ethyl acetate, freeze-dried to acquire the ethyl acetate layer extract;
( 3) Get the ethyl acetate layer essence as well as dissolve it with 80% methanol remedy (readjust the pH value to 3.5 with acetic acid), and wash it with 80% methanol service (pH worth is 3.5) after removing chlorophyll via the short column of MCl reverse phase chromatography packing Remove until there is no red shade, gather the eluate and concentrate and also completely dry to get the unrefined product of decolorized Theaflavin Supplement;
( 4) Take the unrefined item of the decolorized Theaflavin Supplement, liquify the example in methanol, mix with silica gel by completely dry method, set up a silica gel column, elute with ethyl acetate (containing 2% acetic acid), and accumulate the theaflavin sector by liquid chromatography The eluent is gathered and also incorporated to be focused as well as dried to get the crude item of theaflavin;
( 5) Take the unrefined item of Theaflavin Supplement, dissolve the sample with 40% methanol remedy (including 0.1% acetic acid), and make use of MCl reverse-phase chromatographic column to remove the previous contamination by eluting with 40% methanol solution (including 0.1% acetic acid), and afterwards utilize 60% eluting with methanol solution (consisting of 0.1% acetic acid), collecting the eluate from the theaflavin sector, combining the eluate from the theaflavin segment, concentrating and also drying out to acquire detoxified theaflavin;
( 6) Take the detoxified Theaflavin Supplement sample as well as liquify the example with 30% methanol solution (consisting of 0.1% acetic acid), after that use a C18 reverse-phase chromatographic column to elute with 30% methanol service (including 0.1% acetic acid) to remove the previous pollutants, and then eluted with 50% methanol option (consisting of 0.1% acetic acid), gathered the eluate from the theaflavin sector, integrated the eluate from the theaflavin sector, combined the eluate from the theaflavin sector and also concentrated as well as dried out to acquire a high-purity Yinghong No. 9 black tea theaflavins.
( 1) Prepare 700kg of ethyl acetate option rich in 80mg/ml tea polyphenols by the procedure of removing tea polyphenols;
( 2) 700kg of ethyl acetate option abundant in tea polyphenols and 800kg of shattered fresh tea leaves are gone down into an oxidation fermenter, adhered to by adding 20L of ethyl acetate and 1.5 L of citric acid-disodium hydrogen phosphate barrier, at space temperature level Under the state, the air is gone through to mix and oxidize and ferment for 30-40 mins to get the theaflavin ester stage service;
( 3) The theaflavin ester phase service is first filteringed system to remove the tea dregs, after that concentrated as well as washed with sodium bicarbonate solution to acquire a liquid stage service and also an ester phase remedy; the tea dregs are dried and also stored for later usage.
( 4) Spray-dry after the liquid stage service of action (3) is concentrated, obtain the theaflavin completely dry powder of low spec;
( 5) the ester stage service of action (3) is focused to redeem ethyl acetate, and after that water is contributed to make theaflavin transfer to water;
( 6) The aqueous solution of Theaflavin Supplement after transsolution undergoes centrifugal filtering to remove solid impurities to get a pure theaflavin service;
( 7) Focusing the theaflavin solution to a solid web content of 20-30%, and after that spray drying to acquire a high-standard theaflavin dry powder item.
Via detection and estimation, the purity of the Theaflavin Supplement dry powder product is 55.6%, as well as the return is 54% (determined by tea polyphenols).