( 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.