CN105533019A - Hypoglycemic aglucone-enriched monascus-guava leaf tea as well as preparation method and application thereof - Google Patents
Hypoglycemic aglucone-enriched monascus-guava leaf tea as well as preparation method and application thereof Download PDFInfo
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
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Abstract
The invention discloses a hypoglycemic aglucone-enriched monascus-guava leaf tea as well as a preparation method and an application thereof. The preparation method comprises the following steps: draining the washed guava leaves, crushing the drained guava leaves and then taking the sole crushed guava leaves or a mixture of the crushed guava leaves and natural accessory materials as the culture medium; inoculating the monascus seed liquid into the culture medium to carrying out solid fermentation; drying the culture medium after solid fermentation, so that the hypoglycemic aglucone-enriched monascus-guava leaf tea is prepared. The hypoglycemic aglucone-enriched monascus-guava leaf tea prepared according to the preparation method has a reduced bitter-astringent taste of the guava leaf and an improved fragrant flavor of the tea. According to the preparation method, the macro-molecular functional components of the guava leaf are degraded into micro-molecular functional components; the contents of the aglucones, including quercetin, kaempferol and the like, are increased; and the functional components of the monascus are also increased. Thus, the hypoglycemic aglucone-enriched monascus-guava leaf tea has very large application potential in the field of food or the field of healthcare products.
Description
Technical field
The invention belongs to field of food, particularly a kind of red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon and preparation method and application.
Background technology
Guava Leaf, as a kind of material of medicine-food two-purpose, has use history for many years, have hypoglycemic, anti-inflammatory, antibacterial, anti-oxidant, control dysentery, the multiple curative effect such as cardioprotection.Many research shows that function of polysaccharide composition main in Guava Leaf comprises Flavonoid substances.Its flavonoid substance is based on flavone sugar glycoside material, but the hypoglycemic ability of flavonoid glycoside metaclass material (as Quercetin, Kaempferol etc.) is obviously better than flavone sugar glycoside material (as rutin), and is easier to absorption of human body.Flavone glycoside is converted into the flavone aglycone of the stronger easier absorption of human body of non-oxidizability, tiring of Guava Leaf will be improved greatly.
In addition, Guava Leaf has certain bitter taste.Therefore be that raw material prepares and is rich in flavone aglycone and mouthfeel is easy to the food that allows people accept with Guava Leaf, will greatly improve the economic worth of Guava Leaf.
Summary of the invention
Primary and foremost purpose of the present invention is that the shortcoming overcoming prior art is with not enough, provides a kind of preparation method of red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon.
Another object of the present invention is to the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of the rich hypoglycemic aglycon providing above-mentioned preparation method to obtain.This red colouring agent for food, also used as a Chinese medicine Guava Leaf tea mouthfeel is better, brews soup look ruddy bright.
Another object of the present invention is the application of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea providing described rich hypoglycemic aglycon.
Object of the present invention is achieved through the following technical solutions: a kind of preparation method of red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon, comprises the steps:
(1) drained by the Guava Leaf cleaned up, pulverize, independent Guava Leaf is culture matrix or in Guava Leaf, adds natural auxiliaries and becomes culture matrix; The water content of culture matrix is mass percent 40 ~ 65%;
(2) in culture matrix, inoculate monascus seed liquor, carry out solid state fermentation, ferment to culture matrix for yellow or red to dark violet redness; The condition of solid state fermentation is: the inoculum concentration of monascus seed liquor is 5 ~ 10% (v/w of culture matrix quality, mL/g), in 27 ~ 34 DEG C, 50 ~ 65% humidity bottom fermentations, between yeast phase, the water content of material is made to remain on 40 ~ 65% (w/w) by moisturizing.
(3) culture medium good for solid state fermentation is dried, obtain the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon.
Natural auxiliary material described in step (1) is the composition providing the material of carbon source or provide the material of carbon source and provide the material of nitrogenous source to be formed.
The described material of carbon source that provides is preferably at least one in glucose, sucrose, maltose, corn flour, rice meal and glycerine; Be more preferably rice meal, or the composition of rice meal and glycerine composition.
The described material of nitrogenous source that provides is preferably at least one in peptone, yeast extract and sodium glutamate; Be more preferably peptone.
Described natural auxiliary material is preferably the composition that rice meal, peptone and glycerine are formed; Be more preferably the composition that rice meal, peptone and glycerine are formed by 3g:0.05g:1mL proportioning.
The addition of the natural auxiliary material described in step (1) is preferably equivalent to more than 60% (w/w) of Guava Leaf addition; Be preferably 60 ~ 100%.
Monascus described in step (2) is preferably monascus GIM3.592, but is not limited to this bacterium.
Monascus seed liquor described in step (2) is the monascus ruber being in increased logarithmic phase.
Described monascus seed liquor prepares preferably by following steps: be inoculated in by the monascus ruber activated in sterilized fluid nutrient medium and carry out cultivation propagation in 27 ~ 34 DEG C, make it be in increased logarithmic phase, obtain monascus seed liquor; The formula of fluid nutrient medium is as follows: glucose 20g/L, yeast extract 3g/L, peptone 10g/L, KCl0.05g/L, FeSO
47H
2o0.01g/L and KH
2pO
44g/L, is settled to 1000mL with water.
The monascus ruber that described activation is good prepares preferably by following steps: be inoculated on slant medium by the monascus ruber of preservation, in 27 ~ 34 DEG C of cultivations; The formula of slant medium is glucose 20g/L, yeast extract 3g/L, peptone 10g/L, KCl0.05g/L, FeSO
47H
2o0.01g/L, KH
2pO
44g/L, agar 10 ~ 15g, be settled to 1000mL with water.
Described cultivation propagation preferably expands cultivation step by step.
Described expansion is step by step cultivated and is comprised: use culture vessel to be placed on shaking table and carry out Shaking culture, volume progressively amplifies; Or be first placed on shaking table with culture vessel and carry out Shaking culture, then carry out amplification with bioreactor and cultivate.
The condition that described culture vessel is cultivated is preferably: in culture vessel, the useful load of fluid nutrient medium is equivalent to 1/5 ~ 2/5,27 ~ 34 DEG C of triangular flask volume, 150 ~ 200rpm cultivates 27 ~ 28h.
Described culture vessel is preferably triangular flask.
The condition of described bioreactor is preferably: in bioreactor, the useful load of fluid nutrient medium is equivalent to 50 ~ 80% of bioreactor volume, 27 ~ 34 DEG C of aerobic cultivations.
Preferably 8 ~ 16 days time of the solid state fermentation described in step (2); Be more preferably 13 ~ 15 days.
The condition of the moisturizing described in step (2) is preferably moisturizing in the 7th day of solid state fermentation, and rate of water make-up is 2mL, thus makes water content remain on 40 ~ 65% (w/w).
The temperature of the oven dry described in step (3) is preferably 50 ~ 60 DEG C.
A red colouring agent for food, also used as a Chinese medicine Guava Leaf tea for rich hypoglycemic aglycon, is obtained by above-mentioned preparation method.
The red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of described rich hypoglycemic aglycon is applied in field of food and/or field of health care products; It is direct-edible, also can be processed into varieties of food items further, as the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea biscuit etc. of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea beverage of rich hypoglycemic aglycon, rich hypoglycemic aglycon.
The present invention has following advantage and effect relative to prior art:
The red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of the rich hypoglycemic aglycon obtained by preparation method provided by the invention reduces the bitter taste of Guava Leaf, increase tea smell, improve and brew soup look, and be Small molecular functional component by the large molecular function ingredient degradation of Guava Leaf, improve Quercetin, the Aglycones content such as Kaempferol, add red colouring agent for food, also used as a Chinese medicine functional component simultaneously, include but not limited to monascorubin, citrinin (Monacolin) class material, GABA, the compositions such as ergosterol, these compositions are to the oxidation resistance strengthening body, reduce cholesterol, prevention cardiovascular and cerebrovascular disease, improve constipation etc. and there is facilitation.
Accompanying drawing explanation
Fig. 1 is that embodiment 1 adds Quercetin in the red colouring agent for food, also used as a Chinese medicine Guava Leaf of different carbon source ferment making, Kaempferol changes of contents testing result figure; Wherein: M is monascus, CK is the Guava Leaf before not fermenting.
Fig. 2 is that embodiment 1 adds pigment content change testing result figure in the red colouring agent for food, also used as a Chinese medicine Guava Leaf of different carbon source ferment making; Wherein: M monascus, CK is the Guava Leaf before not fermenting.
Fig. 3 is that embodiment 2 adds Quercetin in the red colouring agent for food, also used as a Chinese medicine Guava Leaf of different nitrogen sources ferment making, Kaempferol changes of contents testing result figure; Wherein: M is monascus, CK is the Guava Leaf before not fermenting.
Fig. 4 is that embodiment 2 adds pigment content change testing result figure in the red colouring agent for food, also used as a Chinese medicine Guava Leaf of different egg sources ferment making; Wherein: M is monascus, CK is the Guava Leaf before not fermenting.
Fig. 5 is that embodiment 3 adds Quercetin, Kaempferol changes of contents testing result figure in the red colouring agent for food, also used as a Chinese medicine Guava Leaf of different carbon source, nitrogenous source orthogonal experiment optimal set ferment making; Wherein: M is monascus, CK is the Guava Leaf before not fermenting.
Fig. 6 is that embodiment 3 adds pigment content change testing result figure in the red colouring agent for food, also used as a Chinese medicine Guava Leaf of different carbon source, nitrogenous source orthogonal test optimal set ferment making; Wherein: M is monascus, CK is the Guava Leaf before not fermenting.
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment 1
(1) preparation of seed liquor: adopt Monascus GIM3.592 (MonascusankaGIM3.592, Guangdong Province's Culture Collection).Be inoculated in fluid nutrient medium by the inclined-plane seed activated and cultivate, inoculum concentration is (5 ~ 10) × 10
5individual cell/mL, 29 ~ 32 DEG C, 180rpm shaking table cultivation 28 ~ 32h, make seed liquor have ripe spore, obtain seed liquor; The formula of liquid seed culture medium is as follows: glucose 20g/L, yeast extract 3g/L, peptone 10g/L, KCl0.05g/L, FeSO
47H
2o0.01g/L and KH
2pO
44g/L, is settled to 1000ml with water.
(2) solid state fermentation: add 5g Guava Leaf (this Guava Leaf is through cleaning, draining, pulverize) in 250ml triangular flask, add 3g sugar (glucose, sucrose or maltose) or 3g starchy material (corn flour or rice meal) (all for providing carbon source), initial water content 55%, gross weight is about 20g.121 DEG C of sterilizing 20min.After culture medium cooling, the seed liquor that inoculation step (1) prepares in this solid-state fermentation culture medium, inoculum concentration is 10% (v/w) of solid-state fermentation culture medium gross mass; Fermentation is placed 15 days at 30 DEG C.Ferment moisturizing in the 7th day once, rate of water make-up 2mL, makes the water content of material remain on 45 ~ 60% (w/w).
(3) after having fermented then 60 DEG C dry 15h, obtain the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon.Pulverize, by 40 mesh sieves, for following composition detection with pulverising mill:
1. the detection of citrinin: get 2.0g sample in 100mL centrifuge tube, adds 50mL70% (v/v) ethanol, and ultrasonic (80w) extracts 30min, and 0.45 μm of Filter paper filtering is got supernatant and carry out HPLC analysis after micropore organic membrane filter.Concrete analysis condition is: the highly effective liquid phase chromatographic system of fluorescence detector, determined wavelength-excitation wavelength 331nm, emission wavelength 500nm, column temperature 30 DEG C, C18 chromatographic column.Mobile phase used is: A-water: 36% acetic acid=100:10 (v:v), B-acetonitrile: 36% acetic acid=100:10 (v:v), and flow velocity is 1.0mL/min, sample size 20 μ L.Testing conditions: gradient elution-0min, 80%A+20%B, 28min, 50%A+50%B, 30min, 15%A+85%B, 50min, 15%A+85%B (being volume ratio).
2. the detection of Monacolin K: accurately take sample 0.4g, in 50mL volumetric flask, adds 75% (v/v) ethanol constant volume, ultrasonic under room temperature (80w) 1h.With the centrifugal 10min of 3500r/min rotating speed, get supernatant and cross organic miillpore filter, filtrate is analyzed through HPLC.Concrete analysis condition is: the highly effective liquid phase chromatographic system of UV-detector, determined wavelength 232nm, column temperature 30 DEG C, C18 chromatographic column.Mobile phase used is: A-pH=2.5 ultra-pure water (phosphoric acid adjusts pH), B-acetonitrile, flow velocity is 1.0ml/min, sample size 10 μ L.Testing conditions: 45% (v/v) pH2.5 (phosphoric acid adjusts pH) ultra-pure water, 55% (v/v) acetonitrile.Analysis time is 50min.
3. the detection of pigment: take 0.1g sample and be placed in 50mL tool grinding port plug test tube with a scale, add and adjust 70% ethanol (v/v) of pH=2 to 50mL with hydrochloric acid, 60 DEG C of water-bath 1h are put into after shaking up, filtrate is diluted suitable multiple by the ethanolic solution taking out the pH=2 of the rear comparable sodium of cooling, under wavelength 410nm, 470nm and 510nm, red colouring agent for food, also used as a Chinese medicine Guava Leaf dark brown (light absorption value) AU410 is measured with ultraviolet-uisible spectrophotometer, AU470 and AU510 is worth, this value is multiplied by extension rate and is uranidin, citraurin, red plain color valency, unit is AU/g.
4. Quercetin and kaempferol detect: accurately take sample 0.4g, in 10mL volumetric flask, add 75% (v/v) ethanol constant volume, ultrasonic 30min under room temperature.0.45 μm of Filter paper filtering, get clear liquid and cross organic miillpore filter, filtrate is analyzed through HPLC.Concrete analysis condition is: the highly effective liquid phase chromatographic system of UV-detector, determined wavelength 370nm, column temperature 30 DEG C, C18 chromatographic column.Mobile phase used is: A-1% aqueous acetic acid, B-methyl alcohol: acetonitrile=40:15 (adding 1% acetic acid), and flow velocity is 0.5mL/min, sample size 10 μ L.Testing conditions: gradient elution-0min, 68%A+32%B, 22min, 68%A+32%B, 50min, 23.2%A+76.8%B, 55 ~ 60min, 68%A+32%B (being volume ratio).Analysis time is 60min.
According to the rich hypoglycemic aglycon red koji fermentation Guava Leaf tea that above-mentioned condition is obtained, relatively non-fermented tea and red koji fermentation Guava Leaf tea brew soup look, it is bright that visible red koji fermentation Guava Leaf tea brews soup look ruddy, it is pinkish that unfermentable Guava Leaf brews soup look, and red koji fermentation Guava Leaf liquor color is more tempting.
Adopt high performance liquid chromatography to detect Quercetin and Kaempferol content, as shown in Figure 1, when wherein adding rice, the content of Quercetin and Kaempferol is higher, is respectively 1.396mg/g, 0.204mg/g for result.After testing, haematochrome content is 420AU/g, and citraurin content is 440AU/g, and Yellow pigment content is 785AU/g, the results are shown in Figure 2.Contrast the contrast of unfermentable Guava Leaf by the product after monascus+rice meal fermentation, find that the content of wherein newly-increased composition Mo Nakelin K is 0.45mg/g, citrinin do not detected.Detect data in table 1.
Embodiment 2
(1) preparation of seed liquor: with embodiment 1 step (1).
(2) solid state fermentation: add 5g Guava Leaf (this Guava Leaf is by cleaning, draining, pulverize) in 250ml triangular flask, add 3g rice meal (for providing carbon source) and 0.05g nitrogenous source (peptone, yeast extract or sodium glutamate), initial water content 55%.121 DEG C of sterilizing 20min.After culture medium cooling, the seed liquor that inoculation step (1) prepares in this solid-state fermentation culture medium, inoculum concentration is 10% (v/w) of solid-state fermentation culture medium quality; Fermentation is placed 15 days at 30 DEG C.Ferment moisturizing in the 7th day once, rate of water make-up 2mL, makes the water content of material remain on 45 ~ 60% (w/w).
(3) after having fermented then 60 DEG C dry 15h, obtain the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon.Detection method is with embodiment 1.
According to the rich hypoglycemic aglycon red koji fermentation Guava Leaf tea that above-mentioned condition is obtained, adopt high performance liquid chromatography to detect Quercetin and Kaempferol content, the results are shown in Figure 3, when wherein adding 0.25% peptone, the content of Quercetin and Kaempferol is higher is respectively 1.892mg/g, 0.218mg/g.After testing, haematochrome content is 380AU/g, and citraurin content is 420AU/g, and Yellow pigment content is 760AU/g, the results are shown in Figure 4.Relative to unfermentable Guava Leaf, wherein the content of newly-increased composition Mo Nakelin K is 0.23mg/g, citrinin do not detected.Detect data in table 1.
Embodiment 3
(1) preparation of seed liquor: with embodiment 1 step (1).
(2) solid state fermentation: pass through rice meal, the orthogonal test (table 2) of peptone and glycerine addition draws, 5g Guava Leaf, rice meal 3g, 0.05g peptone and 1mL glycerine is added, initial water content 55% (gross mass is about 20g) in the triangular flask of 250mL.121 DEG C of sterilizing 20min.After culture medium cooling, the seed liquor that inoculation step (1) prepares in this solid-state fermentation culture medium, inoculum concentration is 10% (v/w) of solid-state fermentation culture medium quality; Fermentation is placed 15 days at 30 DEG C.Ferment moisturizing in the 7th day once, rate of water make-up 2mL, makes the water content of material remain on 45 ~ 60% (w/w).
(3) after having fermented then 60 DEG C dry 15h, obtain the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon.Detection method is with embodiment 1.
According to the rich hypoglycemic aglycon red koji fermentation Guava Leaf tea that above-mentioned condition is obtained, high performance liquid chromatography is adopted to detect Quercetin and Kaempferol content, the results are shown in Figure 5, wherein gross mass is about in 20g culture medium and adds 3g rice meal, 0.05g peptone, during 1mL glycerine, the content best result of Quercetin and Kaempferol Wei 1.895mg/g, 0.343mg/g.After testing, haematochrome content is 440AU/g, and citraurin content is 430AU/g, and Yellow pigment content is 750AU/g, the results are shown in Figure 6.Wherein the content of newly-increased composition Mo Nakelin K is 0.43mg/g, citrinin do not detected.Detect data in table 1.
Functional component testing result in each example of table 1
Table 2 example three orthogonal table
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from Spirit Essence of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. a preparation method for the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon, is characterized in that comprising the steps:
(1) drained by the Guava Leaf cleaned up, pulverize, independent Guava Leaf is culture matrix or in Guava Leaf, adds natural auxiliaries and becomes culture matrix; The water content of culture matrix is mass percent 40 ~ 65%;
(2) in culture matrix, inoculate monascus seed liquor, carry out solid state fermentation, ferment to culture matrix for yellow or red to dark violet redness; The condition of solid state fermentation is: the inoculum concentration of monascus seed liquor is 5 ~ 10% of culture matrix quality, in 27 ~ 34 DEG C, 50 ~ 65% humidity bottom fermentations, makes the water content of material remain on 40 ~ 65% between yeast phase by moisturizing;
(3) culture medium good for solid state fermentation is dried, obtain the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon;
Natural auxiliary material described in step (1) is the composition providing the material of carbon source or provide the material of carbon source and provide the material of nitrogenous source to be formed.
2. the preparation method of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon according to claim 1, is characterized in that:
The described material of carbon source that provides is at least one in glucose, sucrose, maltose, corn flour, rice meal and glycerine;
The described material of nitrogenous source that provides is at least one in peptone, yeast extract and sodium glutamate.
3. the preparation method of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon according to claim 2, is characterized in that: the composition that described natural auxiliary material rice meal, peptone and glycerine are formed.
4. the preparation method of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon according to claim 1, is characterized in that: the addition of the natural auxiliary material described in step (1) is more than 60% (w/w) being equivalent to Guava Leaf addition.
5. the preparation method of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon according to claim 1, is characterized in that: the monascus seed liquor described in step (2) is the monascus ruber being in increased logarithmic phase.
6. the preparation method of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon according to claim 5, it is characterized in that: described monascus seed liquor prepares as follows: the monascus ruber activated is inoculated in sterilized fluid nutrient medium and carries out cultivation propagation in 27 ~ 34 DEG C, make it be in increased logarithmic phase, obtain monascus seed liquor; The formula of fluid nutrient medium is as follows: glucose 20g/L, yeast extract 3g/L, peptone 10g/L, KCl0.05g/L, FeSO
47H
2o0.01g/L and KH
2pO
44g/L, is settled to 1000mL with water.
7. the preparation method of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon according to claim 6, is characterized in that:
The monascus ruber that described activation is good prepares as follows: be inoculated on slant medium by the monascus ruber of preservation, in 27 ~ 34 DEG C of cultivations; The formula of slant medium is glucose 20g/L, yeast extract 3g/L, peptone 10g/L, KCl0.05g/L, FeSO
47H
2o0.01g/L, KH
2pO
44g/L, agar 10 ~ 15g, be settled to 1000mL with water;
Described cultivation propagation is for expand cultivation step by step.
8. the preparation method of the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon according to claim 1, is characterized in that:
The time of the solid state fermentation described in step (2) is 8 ~ 16 days;
The condition of the moisturizing described in step (2) is moisturizing in the 7th day of solid state fermentation, and rate of water make-up is 2mL, thus makes water content remain on 40 ~ 65% (w/w).
9. a red colouring agent for food, also used as a Chinese medicine Guava Leaf tea for rich hypoglycemic aglycon, be is characterized in that: obtained by the preparation method described in any one of claim 1 ~ 8.
10. the red colouring agent for food, also used as a Chinese medicine Guava Leaf tea of rich hypoglycemic aglycon according to claim 9 is applied in field of food or field of health care products.
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Cited By (1)
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| CN111772015A (en) * | 2020-07-17 | 2020-10-16 | 浙江红石梁集团天台山乌药有限公司 | Red yeast black-bone leaf tea, granules and preparation method of beverage of red yeast black-bone leaf tea |
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Application publication date: 20160504 |