CN107118920A

CN107118920A - High flavonoids applejack of the strong type of sweet tea and preparation method thereof

Info

Publication number: CN107118920A
Application number: CN201710524264.0A
Authority: CN
Inventors: 陈学森; 左卫芳; 张天亮; 鲁墨森; 张宗营; 王楠; 姜生辉; 许海峰; 王意程; 毛志泉; 姜远茂
Original assignee: Shandong Agricultural University
Current assignee: Shandong Agricultural University
Priority date: 2017-06-30
Filing date: 2017-06-30
Publication date: 2017-09-01
Anticipated expiration: 2037-06-30
Also published as: CN107118920B

Abstract

The invention discloses high flavonoids applejack of the strong type of a kind of sweet tea and preparation method thereof.The high sugar wine base of 1 parts by volume and the low wine base of 6 parts by volume are mixed to get low high sugar wine base.The ripening fruits of height wine base, low wine base and high sugar wine base using high flavonoids apple is raw material.Height wine base preparation method：Fermentation no longer raises to alcoholic strength, distills 85 95 DEG C of distillates of collection, until alcoholic strength is more than 80%；Low wine base preparation method：Fermentation no longer raises to alcoholic strength, collects liquid phase；High sugar wine base preparation method：Fermentation to alcoholic strength is 1%, sterilizing, collects liquid phase.The high flavonoids apple method for preparing medicated wine of the strong type of sweet tea：1 parts by volume height wine base is mixed with the low high sugar wine base of 0.5 parts by volume, and then high and low temperature alternative is cured.The high flavonoids applejack of the strong type of sweet tea that the present invention is provided, both containing the distinctive functional health composition of the apples such as flavonoids, phenolic acid and mineral element, has stronger excitant and white wine mouthfeel, with huge market prospect again.

Description

High flavonoids applejack of the strong type of sweet tea and preparation method thereof

Technical field

The present invention relates to high flavonoids applejack of the strong type of a kind of sweet tea and preparation method thereof.

Background technology

The number of degrees of wine, also known as alcoholic strength, refer to the percent by volume shared by ethanol in wine, unless otherwise specified, generally Refer to the percent by volume in wine shared by ethanol at 20 DEG C.Alcoholic strength typically represents with x%, sometimes also with x% (V/V) or X%vol or x ° of expression.Alcoholic strength is that x% is meant, at 20 DEG C, the second containing x unit volumes in the wine of 100 unit volumes Alcohol.

Low alcohol fruit wine, such as claret, dry white wine, due to containing flavonoids (anthocyanin), needed by human The nutritive and health protection components such as amino acid and mineral element, are in recent years obvious rising situation in the consumption of China.But, low fruit The alcohol content of wine is low, stimulation is small, it is impossible to effectively meet the demand that custom drinks the crowd of white wine.Drink white to meet custom The demand of wine crowd, generally increases low alcohol fruit wine by way of the sugaring in zymotic fluid or edible alcohol in the prior art Alcoholic strength.

Wine and spirits culture, are Chinese diet and socio-cultural important component.White wine has been permeated in whole China five In the civilized history of thousand, occupy important from literary and artistic creation, entertainment to everyways such as diet culinary art, health cares Position.Visitor comes from a distant place, and no wine is not enough to express deep feeling kindness.In the fine moment happy festival time, it is cheerful and light-hearted satisfied that no wine is not enough to display.Spring breeze is obtained Meaning, no wine is not enough to the lofty ideal that gives off one's lofty sentiments.Therefore, although low alcohol fruit wine is in obvious rising situation in the consumption of China, have The white wine of certain wine degree will still have very big development space in future.

" doctor's food homology, eats nutrition, eats health " turns into the common recognition of people.Apple contains more human body and easily inhaled Receive free polyphenol or flavonoids, it is anti-oxidant, prevention cardiovascular and cerebrovascular disease and it is antitumor in terms of be respectively provided with preferable effect, And phenolic acid (Phenolicacids) is the organic acid that a class contains phenol ring, with arousing brain, raising spirit, enriching yin beauty treatment, skin whitening, clear Be pyrolyzed fire effect, " one day apple, doctor is away from me ", in the world it is considerable country all using apple be used as major consumers fruit Product and recommend energetically.But for a long time, due to the undue pursuit to yield and outward appearance, and some merits such as fruit polyphenol exist The long korneforos of history is gradually eliminated.Therefore, seminar where Chen Xuesen professor takes the lead in proposing " functional form (high flavonoids) apple Concept really " and its breeding strategy, to improve breeding of new variety scheme, improve breeding efficiency, expand answering for high flavonoids apple With scope, four main measures are taken：One is to property such as Xinjiang red meat apple and apple variety first generation of hybrid fruit total phenol contents On the basis of shape Research on Genetic Variation, the Apple breeding method (patent No. ZL 2,013 1 of " three select a two early rush " is proposed and implemented 0205419.6), breeding efficiency is significantly improved；Two be to utilize the apple varieties such as genetic background complicated ' loud, high-pitched sound ' and Xinjiang red meat Apple (M.sieversiif.niedzwetzkyana) carries out multi-parent strain hybridization with being returned repeatedly, it is intended to carry out quality breeding, mesh It is preceding to have built miscellaneous (time) friendship one, two generation segregating population 40,40,000 plants of hybrid seedling is colonized, and declared " many introduces a collection product of fruit tree Matter method of breeding " (number of patent application 2,015 10428448.8) and " easy coloring apple variety cultivating method " (number of patent application 201510890141.X) 2 breeding technique patents of invention；Three be in time using the basicly stable offspring's strain of character as examination material, Quality trait evaluation and the research of development mechanism are carried out, and has achieved many impressive progresses；Four be according to " removable, small-sized Change, it is intelligent, pure natural, no added " theory, have developed including the high flavonoids apple crushing and beating device of a kind of stroke type, many High flavonoids applejack complete equipment for processing and the processing works such as function wine brewing fractionator, the roasting drinking vessel of multiple-effect condensation and separation Wine storage device Skill.At present, the apple high-efficient breeding technique system that conventional hybridization is organically combined with biotechnology has been created, has formulated a collection of new Kind and excellent germplasm, have developed Variety of Apple highly effective matched cultivation and deep process technology system.Authorize and declare invention special Sharp 30 remainders, are bred as new varieties (being) 16；Correlative study paper 120, the wherein piece of SCI papers more than 20 are delivered, these researchs Achievement is totally in the top standard of international similar research.

The content of the invention

It is an object of the invention to provide high flavonoids applejack of the strong type of a kind of sweet tea and preparation method thereof.

Present invention firstly provides a kind of composition for being used to produce applejack, including height wine base and low high sugar wine Base；

The low wine base of the high sugar wine base of 1 parts by volume and 5.5-6.5 parts by volume is mixed to get by the low high sugar wine base；

The height wine base, the low wine base and the high sugar wine base are using the ripening fruits of high flavonoids apple as original Material；

The preparation method of the height wine base in turn includes the following steps：

(1) crushing and beating；

(2) sterilize；

(3) pectase is added；

(4) add saccharomyces cerevisiae and fermented, until the alcoholic strength of fermentation system is no longer raised；

(5) distill and collect the distillate between 85-95 DEG C, then distillate is distilled again and collect 85-95 DEG C it Between distillate, repeatedly, until obtaining the distillate that alcoholic strength is more than 80%, as height wine base；

The preparation method of the low wine base in turn includes the following steps：

(1) crushing and beating；

(2) sterilize；

(3) pectase is added；

(5) liquid phase is collected, is low wine base；

The preparation method of the high sugar wine base in turn includes the following steps：

(1) crushing and beating；

(2) sterilize；

(3) pectase is added；

(4) add saccharomyces cerevisiae and fermented, until the alcoholic strength of fermentation system is 0.8%-1.2%；

(5) sterilize；

(6) liquid phase, as high sugar wine base are collected.

The low wine base of the high sugar wine base of 1 parts by volume and 6 parts by volume is mixed to get by the low high sugar wine base.

In the preparation method of the height wine base, the ripening fruits of the high flavonoids apple is unique raw material.The height In the preparation method for spending wine base, the ripening fruits of the high flavonoids apple is the fruit after cleaning.The system of the height wine base In Preparation Method, the ripening fruits of the high flavonoids apple is the fruit after being cleaned with running water.The preparation of the height wine base In method, the ripening fruits of the high flavonoids apple is with the fruit after running water thoroughly cleaning impurity elimination.

In the step (1) of the preparation method of the height wine base, crushing and beating to pulp granularity is below 1mm.

In the step (2) of the preparation method of the height wine base, the condition of the sterilizing is：80-100 DEG C of sterilizing 5- 12 hours.In the step (2) of the preparation method of the height wine base, the condition of the sterilizing is concretely：100 DEG C, it is 5 small When.

In the step (3) of the preparation method of the height wine base, when pulp temperature is 45-55 DEG C, pectin is added Enzyme.In the step (3) of the preparation method of the height wine base, when it is 45-55 DEG C that pulp, which naturally cools to temperature, add Pectase.In the step (3) of the preparation method of the height wine base, when pulp temperature is 45-50 DEG C, pectin is added Enzyme.In the step (3) of the preparation method of the height wine base, when it is 45-50 DEG C that pulp, which naturally cools to temperature, add Pectase.In the step (3) of the preparation method of the height wine base, when pulp temperature is 45 DEG C, pectase is added.Institute In the step (3) for the preparation method for stating height wine base, when it is 45 DEG C that pulp, which naturally cools to temperature, pectase is added. In the step (3) of the preparation method of the height wine base, the addition of the pectase is：Every kilogram high flavonoids apple Ripening fruits accordingly add 1,000,000 more than u pectase.In the step (3) of the preparation method of the height wine base, institute The addition for stating pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight accordingly add the u of 1,000,000 u-150 ten thousand with On pectase.In the step (3) of the preparation method of the height wine base, the addition of the pectase is：Every kilogram The ripening fruits of high flavonoids apple accordingly adds the u of 1,000,000 u-150 ten thousand pectase.The preparation method of the height wine base In the step (3), the addition of the pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight is accordingly added The u of 1000000 u-150 ten thousand pectase.In the step (3) of the preparation method of the height wine base, the addition of the pectase Measure and be：The ripening fruits of every kilogram high flavonoids apple accordingly adds 2-3g pectases.The preparation method of the height wine base In the step (3), the addition of the pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight is accordingly added 2-3g pectases.

In the step (4) of the preparation method of the height wine base, when pulp temperature is 25-35 DEG C, wine brewing is added Yeast.In the step (4) of the preparation method of the height wine base, when it is 25-35 DEG C that pulp, which naturally cools to temperature, plus Enter saccharomyces cerevisiae.In the step (4) of the preparation method of the height wine base, when pulp temperature is 30 DEG C, wine brewing is added Yeast.In the step (4) of the preparation method of the height wine base, when it is 30 DEG C that pulp, which naturally cools to temperature, add Saccharomyces cerevisiae.In the step (4) of the preparation method of the height wine base, the addition of the saccharomyces cerevisiae is：Every kilogram The ripening fruits of high flavonoids apple accordingly adds 10¹⁰Cfu saccharomyces cerevisiaes.The step of the preparation method of the height wine base (4) in, the addition of the saccharomyces cerevisiae is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight accordingly adds 10¹⁰cfu Saccharomyces cerevisiae.During the fermentation, the temperature of fermentation system, control are continued to monitor since adding after saccharomyces cerevisiae 24 hours Temperature processed is 15-20 DEG C.When described " until the alcoholic strength of fermentation system is no longer raised ", the alcoholic strength of fermentation system is 9%- 12%.The saccharomyces cerevisiae concretely high flavonoids saccharomyces cidri one.

It is described " until it is more than 80% to obtain alcoholic strength in the step (5) of the preparation method of the height wine base Distillate ", concretely obtains the distillate that alcoholic strength is 80%-85%.The step of the preparation method of the height wine base Suddenly in (5), described " until obtaining the distillate that alcoholic strength is more than 80% " concretely obtains alcoholic strength and distillated for 80% Thing.

In the preparation method of the low wine base, the ripening fruits of the high flavonoids apple is unique raw material.It is described low In the preparation method for spending wine base, the ripening fruits of the high flavonoids apple is the fruit after cleaning.The system of the low wine base In Preparation Method, the ripening fruits of the high flavonoids apple is the fruit after being cleaned with running water.The preparation of the low wine base In method, the ripening fruits of the high flavonoids apple is with the fruit after running water thoroughly cleaning impurity elimination.

In the step (1) of the preparation method of the low wine base, crushing and beating to pulp granularity is below 1mm.

In the step (2) of the preparation method of the low wine base, the condition of the sterilizing is：80-100 DEG C of sterilizing 5- 12 hours.In the step (2) of the preparation method of the low wine base, the condition of the sterilizing is concretely：100 DEG C, it is 5 small When.

In the step (3) of the preparation method of the low wine base, when pulp temperature is 45-55 DEG C, pectin is added Enzyme.In the step (3) of the preparation method of the low wine base, when it is 45-55 DEG C that pulp, which naturally cools to temperature, add Pectase.In the step (3) of the preparation method of the low wine base, when pulp temperature is 45-50 DEG C, pectin is added Enzyme.In the step (3) of the preparation method of the low wine base, when it is 45-50 DEG C that pulp, which naturally cools to temperature, add Pectase.In the step (3) of the preparation method of the low wine base, when pulp temperature is 45 DEG C, pectase is added.Institute In the step (3) for the preparation method for stating low wine base, when it is 45 DEG C that pulp, which naturally cools to temperature, pectase is added. In the step (3) of the preparation method of the low wine base, the addition of the pectase is：Every kilogram high flavonoids apple Ripening fruits accordingly add 1,000,000 more than u pectase.In the step (3) of the preparation method of the low wine base, institute The addition for stating pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight accordingly add the u of 1,000,000 u-150 ten thousand with On pectase.In the step (3) of the preparation method of the low wine base, the addition of the pectase is：Every kilogram The ripening fruits of high flavonoids apple accordingly adds the u of 1,000,000 u-150 ten thousand pectase.The preparation method of the low wine base In the step (3), the addition of the pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight is accordingly added The u of 1000000 u-150 ten thousand pectase.In the step (3) of the preparation method of the low wine base, the addition of the pectase Measure and be：The ripening fruits of every kilogram high flavonoids apple accordingly adds 2-3g pectases.The preparation method of the low wine base In the step (3), the addition of the pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight is accordingly added 2-3g pectases.

In the step (4) of the preparation method of the low wine base, when pulp temperature is 25-35 DEG C, wine brewing is added Yeast.In the step (4) of the preparation method of the low wine base, when it is 25-35 DEG C that pulp, which naturally cools to temperature, plus Enter saccharomyces cerevisiae.In the step (4) of the preparation method of the low wine base, when pulp temperature is 30 DEG C, wine brewing is added Yeast.In the step (4) of the preparation method of the low wine base, when it is 30 DEG C that pulp, which naturally cools to temperature, add Saccharomyces cerevisiae.In the step (4) of the preparation method of the low wine base, the addition of the saccharomyces cerevisiae is：Every kilogram The ripening fruits of high flavonoids apple accordingly adds 10¹⁰Cfu saccharomyces cerevisiaes.The step of the preparation method of the low wine base (4) in, the addition of the saccharomyces cerevisiae is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight accordingly adds 10¹⁰cfu Saccharomyces cerevisiae.During the fermentation, the temperature of fermentation system, control are continued to monitor since adding after saccharomyces cerevisiae 24 hours Temperature processed is 15-20 DEG C.When described " until the alcoholic strength of fermentation system is no longer raised ", the alcoholic strength of fermentation system is 9%- 12%.The saccharomyces cerevisiae concretely high flavonoids saccharomyces cidri one.

In the step (5) of the preparation method of the low wine base, " the collection liquid phase " concretely " crosses 600 mesh Sieve, collects liquid phase ".

In the preparation method of the high sugar wine base, the ripening fruits of the high flavonoids apple is unique raw material.The height In the preparation method of sugar wine base, the ripening fruits of the high flavonoids apple is the fruit after cleaning.The system of the high sugar wine base In Preparation Method, the ripening fruits of the high flavonoids apple is the fruit after being cleaned with running water.The preparation of the high sugar wine base In method, the ripening fruits of the high flavonoids apple is with the fruit after running water thoroughly cleaning impurity elimination.

In the step (1) of the preparation method of the high sugar wine base, crushing and beating to pulp granularity is below 1mm.

In the step (2) of the preparation method of the high sugar wine base, the condition of the sterilizing is：80-100 DEG C of sterilizing 5- 12 hours.In the step (2) of the preparation method of the high sugar wine base, the condition of the sterilizing is concretely：100 DEG C, it is 5 small When.

In the step (3) of the preparation method of the high sugar wine base, when pulp temperature is 45-55 DEG C, pectin is added Enzyme.In the step (3) of the preparation method of the high sugar wine base, when it is 45-55 DEG C that pulp, which naturally cools to temperature, add Pectase.In the step (3) of the preparation method of the high sugar wine base, when pulp temperature is 45-50 DEG C, pectin is added Enzyme.In the step (3) of the preparation method of the high sugar wine base, when it is 45-50 DEG C that pulp, which naturally cools to temperature, add Pectase.In the step (3) of the preparation method of the high sugar wine base, when pulp temperature is 45 DEG C, pectase is added.Institute In the step (3) for the preparation method for stating high sugar wine base, when it is 45 DEG C that pulp, which naturally cools to temperature, pectase is added. In the step (3) of the preparation method of the high sugar wine base, the addition of the pectase is：Every kilogram high flavonoids apple Ripening fruits accordingly add 1,000,000 more than u pectase.In the step (3) of the preparation method of the high sugar wine base, institute The addition for stating pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight accordingly add the u of 1,000,000 u-150 ten thousand with On pectase.In the step (3) of the preparation method of the high sugar wine base, the addition of the pectase is：Every kilogram The ripening fruits of high flavonoids apple accordingly adds the u of 1,000,000 u-150 ten thousand pectase.The preparation method of the high sugar wine base In the step (3), the addition of the pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight is accordingly added The u of 1000000 u-150 ten thousand pectase.In the step (3) of the preparation method of the high sugar wine base, the addition of the pectase Measure and be：The ripening fruits of every kilogram high flavonoids apple accordingly adds 2-3g pectases.The preparation method of the high sugar wine base In the step (3), the addition of the pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight is accordingly added 2-3g pectases.

In the step (4) of the preparation method of the high sugar wine base, when pulp temperature is 25-35 DEG C, wine brewing is added Yeast.In the step (4) of the preparation method of the high sugar wine base, when it is 25-35 DEG C that pulp, which naturally cools to temperature, plus Enter saccharomyces cerevisiae.In the step (4) of the preparation method of the high sugar wine base, when pulp temperature is 30 DEG C, wine brewing is added Yeast.In the step (4) of the preparation method of the high sugar wine base, when it is 30 DEG C that pulp, which naturally cools to temperature, add Saccharomyces cerevisiae.In the step (4) of the preparation method of the high sugar wine base, the addition of the saccharomyces cerevisiae is：Every kilogram The ripening fruits of high flavonoids apple accordingly adds 10¹⁰Cfu saccharomyces cerevisiaes.The step of the preparation method of the high sugar wine base (4) in, the addition of the saccharomyces cerevisiae is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight accordingly adds 10¹⁰cfu Saccharomyces cerevisiae.During the fermentation, the temperature of fermentation system, control are continued to monitor since adding after saccharomyces cerevisiae 24 hours Temperature processed is 15-20 DEG C.It is described " until the alcoholic strength of fermentation system is 0.8%-1.2% " concretely " until fermentation system Alcoholic strength be 1% ".The saccharomyces cerevisiae concretely high flavonoids saccharomyces cidri one.

In the step (5) of the preparation method of the high sugar wine base, the condition of the sterilizing is：80-100 DEG C of sterilizing 5- 12 hours.In the step (5) of the preparation method of the high sugar wine base, the condition of the sterilizing is concretely：100 DEG C, it is 5 small When.

In the step (6) of the preparation method of the high sugar wine base, " the collection liquid phase " concretely " crosses 600 mesh Sieve, collects liquid phase ".

The present invention also protects application of the composition in applejack is prepared

The present invention also protection one kind prepares the preparation method of applejack (the high flavonoids applejack of the strong type of sweet tea), including following step Suddenly：

1. 1 parts by volume height wine base and the low high sugar wine base of 0.3-0.7 parts by volume are mixed, first 25~45 DEG C of standings 24~ 72 hours, then 0~4 DEG C stand 24~72 hours, collect liquid phase, be just wine；

2. the first wine that 2. step obtains is taken, high and low temperature alternative curing is carried out, obtains finished wine.

1. the step be：The low high sugar wine base of 1 parts by volume height wine base and 0.5 parts by volume is mixed, first 30 DEG C of standings 24 hours, then 0 DEG C stand 48 hours, collect liquid phase, be just wine.The step 1. in, " the collection liquid phase " concretely " crossing 600 mesh sieves, collect liquid phase ".

The step 2. in, high and low temperature alternative curing is：First 25~45 DEG C stand 24~72 hours, -21 DEG C again~- 15 DEG C stand 24~72 hours, repeat more than 15 times.The step 2. in, high and low temperature alternative curing is：First 30 DEG C Stand and stand 48 hours in 24 hours, again -18 DEG C, repeat 15 times.

The applejack (the high flavonoids applejack of the strong type of sweet tea) that any of the above methods described is prepared falls within the present invention's Protection domain.The number of degrees of the applejack are 52%-58%, concretely 55%.

The high flavonoids apple be every kilogram of fresh weight ripening fruits Flavonoid Content be more than 5000mg apple kind Matter.The high flavonoids apple be every kilogram of fresh weight ripening fruits Flavonoid Content be more than 7000mg apple germplasm.Institute State the apple germplasm that Flavonoid Content that high flavonoids apple is every kilogram of fresh weight ripening fruits is more than 10000mg.The height Flavonoids apple is concretely ' CSR6R6-888 ', ' CSR6R6-666 ' or ' CSR6R6-777 '.

' CSR6R6-888 ', also known as apple (Malus domestica) CSR6R6-888, is protected on June 16th, 2017 Being hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, (abbreviation CGMCC, address is：Chaoyang District, Beijing City The institute 3 of North Star West Road 1, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCC NO.14295.

' CSR6R6-666 ', also known as apple (Malus domestica) CSR6R6-666, was protected on 2 17th, 2017 Being hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, (abbreviation CGMCC, address is：Chaoyang District, Beijing City The institute 3 of North Star West Road 1, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCC NO.13783.

' CSR6R6-777 ', also known as apple (Malus domestica) CSR6R6-777, is protected on December 08th, 2016 Being hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, (abbreviation CGMCC, address is：Chaoyang District, Beijing City The institute 3 of North Star West Road 1, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCC NO.12468.

High flavonoids saccharomyces cidri one, full name is saccharomycete (Saccharomyces sp.) high flavonoids applejack Yeast one, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center's (letter on June 16th, 2017 Claim CGMCC, address is：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica), preservation registration Number be CGMCC NO.14250.

High flavonoids apple excellent germplasm and high flavonoids applejack complete equipment for processing are effectively utilized, developing both has apple Really distinctive functional component, the smell of fruits is very sweet, has stronger excitant and white wine mouthfeel again, can meet different consumer groups' need The serial high flavonoids applejack asked, for elongating Apple Industry chain, rich liquor product Market Diversification, improving enterprise product quality With benefit and meeting and being respectively provided with significance in terms of market and consumption demand.The high flavonoids apple of the strong type of sweet tea that the present invention is provided Fruit wine, both containing the distinctive functional health composition of the apples such as flavonoids, phenolic acid and mineral element, again have stronger excitant and White wine mouthfeel, fruity is fresh and sweet, and the wine is fragrantly scented, and entrance is bold, long times of aftertaste, and the crowd for being adapted to drink high spirit drinks, with weight Big application and popularization value and huge market prospect.

Brief description of the drawings

Fig. 1 is the photo of the high flavonoids applejack of the strong type of ' CSR6R6-888 ' sweet tea.

Fig. 2 is the photo of the high flavonoids applejack of the strong type of ' CSR6R6-888 ' sweet tea.

Embodiment

Following embodiment facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments Method, is conventional method unless otherwise specified.Test material used in following embodiments, is certainly unless otherwise specified What routine biochemistry reagent shop was commercially available.Quantitative test in following examples, is respectively provided with three repetition experiments, as a result makes even Average.

Refer to the bibliography of ' Fuji apple ' apple and ' loud, high-pitched sound ' apple：Chen Xuesen, pungent training just etc., marshal and Jin Shuai are in apple Effect in fruit breeding of new variety, Journal of Shandong agri.Univ, 1994,25 (2)：236—248.

The preparation method of 0.5% methanol hydrochloride solution：The concentrated hydrochloric acid of 0.5 parts by volume 35% is mixed with 99.5 parts by volume methanol Close.

Pectase used is the pectase purchased from Ningxia jade of the He family Bioisystech Co., Ltd, correlation ginseng in embodiment Number is：Solid powder, the u/g of enzyme activity >=500,000, it is proposed that use condition is " pH3.2-5.0,10-50 DEG C ".50.0 DEG C, pH3.5 bars Under part, it is a pectinase activity unit (1u) that 1min catalysis hydrolyzed pectins, which generate the enzyme amount of 1 μ g galacturonic acids,.

The acquisition and identification of embodiment 1, apple excellent germplasm ' CSR6R6-888 '

Identify that the method that apple plants are R1R1 genotype, R6R6 genotype or R6R1 genotype is as follows：From apple to be measured Apple is taken on fruit plant, the genomic DNA of apple pulp is extracted, using genomic DNA as template, the primer constituted using F3 and R3 To entering performing PCR amplification, then by following standard interpretation genotype：If pcr amplification product is a band and is 497bp, to be measured Apple plants are R6R6 genotype；If pcr amplification product is a band and is 386bp, apple plants to be measured are R1R1 genes Type；If pcr amplification product is two bands and respectively 497bp and 386bp, apple plants to be measured are R6R1 genotype.

F3 (sequence 1)：5’-GGTGGTCAAAGATGTGTGTTGT-3’；

R3 (sequence 2)：5’-TTTGCCTGCTACCCACTTCA-3’.

After testing, ' Fuji apple ' apple and ' loud, high-pitched sound ' apple are R1R1 genotype.

First, the acquisition of ' CSR6R6-888 '

Xinjiang red meat apple is as parent, with the plain boiled pork cultivation apple variety hybridization such as ' Fuji apple '.According to Mendelian inheritance Law, the plain boiled pork cultivation apple variety hybridization such as Xinjiang red meat apple (R6R1 genotype) and ' Fuji apple ' (R1R1 genotype), its Progeny population should be red meat phenotype (R6R1 genotype):Plain boiled pork phenotype (R1R1 genotype)=1:1.But, in hybridization F₁Dai Qun The individual plant of R6R6 genotype is found that in body.

The individual plant of one plant of R6R6 genotype is named as ' CSR6R6-888 '.

' CSR6R6-888 ' has following phenotype：The each several parts such as stem, leaf, flower, pericarp and pulp and each stage of development are equal For royal purple.

The fruit fresh food quality of ' CSR6R6-888 '：Sour and sweet palatability, crisp succulence, fresh food quality is excellent.

By way of grafting twig or tissue culture, ' CSR6R6-888 ' is expanded numerous.

2nd, the preservation of ' CSR6R6-888 '

3rd, the preservation of ' CSR6R6-666 '

' CSR6R6-666 ' is the individual plant for another plant of R6R6 genotype that the laboratory early stage where inventor is selected.

' CSR6R6-666 ' has following phenotype：The each several parts such as stem, leaf, flower, pericarp and pulp and each stage of development are equal For aubergine.

4th, the preservation of ' CSR6R6-777 '

' CSR6R6-777 ' is the individual plant for another plant of R6R6 genotype that the laboratory early stage where inventor is selected.

5th, flavonoid component content analysis

' CSR6R6-888 ', ' CSR6R6-666 ' and ' CSR6R6-777 ' is used as plant to be measured respectively.

1st, the transparent apple on plant to be measured is taken, apple pulp is taken.

2nd, the pulp that step 1 is obtained is taken, grinding obtains powder in liquid nitrogen.

3rd, the powder that 2g steps 2 are obtained is weighed, the methanol hydrochloride solutions of 5mL 0.5% are added, 4 DEG C stand extraction 2h, then 8000rpm centrifuges 20min, and supernatant and residue are collected respectively.

4th, the residue that step 3 is obtained is taken, the methanol hydrochloride solutions of 5mL 0.5% are added, 4 DEG C stand extraction 1h, then 8000rpm centrifuges 20min, collects supernatant.

5th, the supernatant mixing that the supernatant and step 4 obtained step 3 is obtained, obtains mixed liquor.

6th, the mixed liquor that step 5 is obtained is taken, 37 DEG C of revolvings remove methanol, and residue 2-3ml methanol dissolves, then 8000rpm centrifuges 20min, collects supernatant.

7th, the supernatant that step 6 is obtained is taken, with methanol constant volume to 5ml, then with 0.45 μm of membrane filtration, filtrate is collected.

8th, the filtrate for obtaining step 7 carries out HPLC-MS analyses.

Liquid phase chromatogram condition：

Using WATERS ACQUITY UPLC chromatographs, chromatographic column is BEH C18 posts (100mm × 2.1mm), filler grain 1.7 μm of footpath；45 DEG C of column temperature；The μ L of sampling volume 1；

Mobile phase is the mixed liquor of A liquid and B liquid, and flow velocity is 0.3mL/min；A liquid is acetonitrile, and B liquid is containing 0.2% (volume Fraction) formic acid the aqueous solution；The volume fraction that 0-0.1min, A liquid account for mobile phase is 5%；0.1-20min, A liquid account for mobile phase Volume fraction is by 5% linear rise to 20%；20-22min, A liquid account for the volume fraction of mobile phase by 20% linear rise extremely 80%；22-22.1min, A liquid account for the volume fraction of mobile phase by 80% linear decline to 5%；22.1-25min, A liquid accounts for flowing The volume fraction of phase is 5%.

Mass Spectrometry Conditions：

Mass spectrograph is WATERS MALDI SYNAPT Q-TOF MS, ESI ionization sources, the collection of electro-spray ionization cation Pattern (ESI+)；Scanning range 100-1500m/z；Capillary voltage 3.5kV, taper hole voltage 30V；100 DEG C of source temperature, precipitation temperature 300 DEG C of degree；Desolventizing gas flow 500L/h.

The content of 9 kinds of specific flavones alcohol matters is shown in Table 1.The detection method of 9 kinds of peculiar materials in table 1 belongs to flavonoids Component and detection method of content, (Chen Xuesen, Zhang Jing, Liu great Liang wait the heredity of the Xinjiang red meat apple first generation of hybrid to bibliography Variation and excellent strain evaluation [J] the Scientia Agricultura Sinicas of functional form apple, 2014,47 (11):2193-2204..

Table 1

Result above shows that it is yellow that ' CSR6R6-888 ', ' CSR6R6-666 ' and ' CSR6R6-777 ' is respectively provided with very strong class Ketone synthesis capability, pulp is rich in flavonoids, and is the excellent of high flavonoids apple variety containing special flavonols component Germplasm.' CSR6R6-888 ' is better than ' CSR6R6-666 ', and ' CSR6R6-666 ' is better than ' CSR6R6-777 '.

6th, Flavonoid Content is determined

(1) the transparent apple fruit on plant to be measured is taken, apple pulp is taken.

(2) 1g pulp is taken, liquid nitrogen grinding, 65% (volumn concentration) ethanol for then adding 4 DEG C of precoolings of 10ml is water-soluble Liquid is simultaneously mixed, and 4 DEG C of lucifuges, which are stood, extracts 4h, and then 12000g centrifuges 20min, collects supernatant.

(3) test tube is taken, the supernatant that 0.5ml steps (2) are obtained is added, then sequentially adds 1mL 5g/100ml NaNO₂ The aqueous solution, 1ml 10g/100ml AL (NO₃)₃The aqueous solution, the 4mL 2M NaOH aqueous solution, stand 15min, 8000rpm after mixing 10min is centrifuged, supernatant is taken, then determines light absorption value under 510nm.

It is that standard specimen does standard curve with rutin (rutin, Sigma chemical, ST, Loiuis, USA).

The Flavonoid Content of the Apple of ' CSR6R6-888 ' is 11358.7mg/kg fresh weights.

The Flavonoid Content of the Apple of ' CSR6R6-666 ' is 9134.6mg/kg fresh weights.

The Flavonoid Content of the Apple of ' CSR6R6-777 ' is 7555.1mg/kg fresh weights.

Embodiment 2, the acquisition of yeast strain

First, the domestication and separation of unartificial yeast

During the ripe apples grown on ' CSR6R6-888 ' plant, excellent apple is selected to take back desinfection chamber in orchard, Pericarp is cut into small pieces in the test tube equipped with fruit juice for being put into and having killed bacterium, is stoppered tampon and is placed in 25~28 DEG C of constant incubators Cultivate 5~8d.Yeast strain is separated using plate streaking partition method, each fermentation liquid does in 3 flat boards, 28 DEG C of insulating boxs and trained Support 3d.Morphologic observation is carried out to bacterium colony, picking separates the good, single bacterium colony with typicalness from flat board, and examination is inoculated in respectively In pipe inclined-plane and sterilized Boiling tube cider, 3 single bacterium colonies are only selected on 3 flat boards of each fermentation liquid, and number Mark, to show bacterium source.Test tube slant is placed in 28 DEG C of culture 3d, checks whether lawn is simple, and the simple good bacterial strain of lawn is put Stored in refrigerator.

2nd, yeast screening assay

1st, Du Shi pipes fermentation screening.

Using Du Shi pipe fermentation methods, under identical condition of culture, the speed of yeast strain aerogenesis bubble is determined and in regulation In time aerogenesis steep number, tentatively more each saccharomycete rise ferment ability and fermentability, filter out fermenting property excellent Yeast strain.Experiment is parallel to be repeated 3 times.Bacterial strain activation condition：25 DEG C of incubated 24h in 10 ° of Brix cider.Hair Ferment condition：20 DEG C of static fermentation 48h in 15 ° of Brix cider.

2nd, saccharomycetes to make fermentation power test (CO₂Weight-loss method).

3rd, saccharomycete coherency compares (yeast number comparison method).

4th, the resistance to ethanol of saccharomycete, resistance to SO₂Experiment.

Using Du Shi pipe fermentation methods, by saccharomycete be respectively connected to different ethanol concentration (6%, 8%, 10%, 12%, V/V) With different SO₂In the cider of concentration (50mg/L, 100mg/L, 150mg/L, 200mg/L), cultivated under the same conditions, The bubble production in Du Shi pipes is observed, relatively more each yeast strain is to ethanol, SO₂Tolerance degree, further determine that suitable The bacterial strain of apple liquor brewing.Experiment is parallel to be repeated 3 times.Bacterial strain activation condition：25 DEG C incubated in 10 ° of Brix cider 24h.Fermentation condition：20 DEG C in 15 ° of Brix cider static fermentation 96h (due to ethanol or SO₂Presence, to saccharomycete Fermentation can produce inhibitory action to varying degrees, therefore aerogenesis observing time is extended for 96h).Inoculum concentration：(6~8) × 10⁷ Individual/ml.

5th, saccharomycete brews cider fermentation experiment.

500ml triangular flasks are taken, 400ml ciders (20 ° of Brix of pol) are added, bacteria concentration is accessed by 20ml/L inoculum concentration For (3~3.6) × 10⁶Individual/ml saccharomycete seed liquor, 20 DEG C of fermentations.Fermentation carries out tank switching to 15d, removes wine pin, then old Make after 10d, determine the physical and chemical index of applejack and carry out organoleptic analysis.

Compared by yeast fermenting power, cohesiveness compares, resistance to SO₂, resistance to ethanol ability compare and it is fermented gained apple The physical and chemical index analysis of wine and organoleptic quality evaluations, filter out three plants of optimal applejack saccharomyces cerevisiaes, are respectively designated as high class yellow Ketone saccharomyces cidri one, high flavonoids saccharomyces cidri two, high flavonoids saccharomyces cidri three.

3rd, three saccharomycetes that comparison step two is obtained produce the performance of applejack

Yeast to be measured is respectively：High flavonoids saccharomyces cidri one, high flavonoids saccharomyces cidri two or high class are yellow Ketone saccharomyces cidri three.

1st, take ' CSR6R6-888 ' without rotten ripening fruits, use running water thoroughly cleaning.

2nd, complete after step 1, take fruit, carry out crushing and beating, until pulp granularity is below 1mm.

3rd, complete after step 2, pulp is sterilized (80 DEG C, 12 hours).

4th, complete after step 3, natural cooling, pectase is added when pulp temperature is 50 DEG C (per the fruit of 1000g fresh weights It is real, add 2-3g pectases).

5th, complete after step 4, continue natural cooling, when pulp temperature is 30 DEG C, add yeast to be measured (fresh per 1000g The fruit of weight, adds 10¹⁰Cfu yeast to be measured), fermented.The alcoholic strength of fermentation system is continued to monitor in fermentation process, when Stop fermentation when alcoholic strength is no longer raised (now the alcoholic strength of system is 9%-12%).In fermentation process, from addition ferment to be measured Start to continue to monitor the temperature of fermentation system after female 24 hours, it is 15-20 DEG C to control temperature.

Physical and chemical index analysis and organoleptic quality evaluations are carried out to obtained applejack, high flavonoids saccharomyces cidri one Indices are most preferable, are resistant to 12% ethanol, and former wine alcoholic strength that it ferments is up to 11.9%, wine body clear, with apple The typical local flavor of fruit wine.

4th, the identification of high flavonoids saccharomyces cidri one

Morphological feature：It is spherical in ellipse, there are obvious nucleus and the vacuole differed in size.

Physiological and biochemical property：Glucose, maltose, fructose, soluble starch, sucrose, galactolipin fermentation production can be utilized Gas, can assimilate glucose, maltose, fructose, soluble starch, sucrose, galactolipin, rhamnose, cellobiose, citric acid, Acid and kind of starch compound can not be produced, ester fragrance matter can be produced.

Growth characteristics：The most suitable growth pH5.5-6.5,29-31 DEG C of optimum growth temperature.

5th, the preservation of high flavonoids saccharomyces cidri one

Embodiment 3 and embodiment 4 are carried out as saccharomyces cerevisiae using high flavonoids saccharomyces cidri one.

The preparation of embodiment 3, height wine base

The apple on the apple on ' CSR6R6-888 ', the apple on ' CSR6R6-666 ' and ' CSR6R6-777 ' is respectively adopted Fruits prepare height wine base.

1st, take without rotten transparent apple fruit, with running water thoroughly cleaning impurity elimination.

3rd, complete after step 2, pulp is sterilized (100 DEG C, 5 hours；Can be using 80-100 DEG C of sterilizing in practical application 5-12 hours, when the temperature is low using longer sterilization time, when temperature is higher using shorter sterilization time, for example when Using the sterilization time of 10-12 hours at 80 DEG C, sterilization time when small using 5 when 100 DEG C).

4th, complete after step 3, natural cooling, (in practical application, 45-55 DEG C can be used, specifically when pulp temperature is 45 DEG C 45-50 DEG C can be used) when, add pectase and (per the fruit of 1000g fresh weights, add 2-3g pectases；The enzyme activity of 2-3g pectases For more than the u of 1,000,000 u-150 ten thousand).

5th, complete after step 4, continue natural cooling, when pulp temperature is 30 DEG C (in practical application, can use 25-35 DEG C) When, add saccharomyces cerevisiae and (per the fruit of 1000g fresh weights, add 10¹⁰Cfu saccharomyces cerevisiaes), fermented.Continue in fermentation process Monitor the alcoholic strength of fermentation system, stop fermentation when alcoholic strength is no longer raised (now the alcoholic strength of system is 9%-12%). In fermentation process, the temperature of fermentation system is continued to monitor since adding after saccharomyces cerevisiae 24 hours, control temperature for 15-20 ℃。

6th, complete after step 5, distilled, collect the distillate between 85-95 DEG C.The alcoholic strength of the distillate is about 20%.

7th, the distillate that step 6 is obtained is taken, is repeatedly distilled, the distillate between 85-95 DEG C is collected every time Distill again, until obtaining the distillate that alcoholic strength is more than 80%, as height wine base.

The height wine base that ' CSR6R6-888 ' is obtained is named as ' CSR6R6-888 ' height wine base.What the present embodiment was obtained The alcoholic strength of ' CSR6R6-888 ' height wine base is 80%.

The height wine base that ' CSR6R6-666 ' is obtained is named as ' CSR6R6-666 ' height wine base.What the present embodiment was obtained The alcoholic strength of ' CSR6R6-666 ' height wine base is 80%.

The height wine base that ' CSR6R6-777 ' is obtained is named as ' CSR6R6-777 ' height wine base.What the present embodiment was obtained The alcoholic strength of ' CSR6R6-777 ' height wine base is 80%.

The preparation of embodiment 4, low high sugar wine base

First, the preparation of low wine base

The apple on the apple on ' CSR6R6-888 ', the apple on ' CSR6R6-666 ' and ' CSR6R6-777 ' is respectively adopted Fruits prepare low wine base.

6th, complete after step 5, cross 600 mesh sieves, collect liquid phase, be low wine base.

The low wine base that ' CSR6R6-888 ' is obtained is named as ' CSR6R6-888 ' low wine base.

The low wine base that ' CSR6R6-666 ' is obtained is named as ' CSR6R6-666 ' low wine base.

The low wine base that ' CSR6R6-777 ' is obtained is named as ' CSR6R6-777 ' low wine base.

2nd, the preparation of high sugar wine base

The apple on the apple on ' CSR6R6-888 ', the apple on ' CSR6R6-666 ' and ' CSR6R6-777 ' is respectively adopted Fruits prepare high sugar wine base.

5th, complete after step 4, continue natural cooling, when pulp temperature is 30 DEG C (in practical application, can use 25-35 DEG C) When, add saccharomyces cerevisiae and (per the fruit of 1000g fresh weights, add 10¹⁰Cfu saccharomyces cerevisiaes), fermented.Continue in fermentation process The alcoholic strength of fermentation system is monitored, fermentation is stopped when alcoholic strength is 1%.In fermentation process, from adding saccharomyces cerevisiae 24 hours Start to continue to monitor the temperature of fermentation system afterwards, it is 15-20 DEG C to control temperature.

6th, complete after step 5, (100 DEG C, 5 hours of heat sterilization；Can be small using 80-100 DEG C of sterilizing 5-12 in practical application When, when the temperature is low using longer sterilization time, when temperature is higher using shorter sterilization time, such as when 80 DEG C Using the sterilization time of 10-12 hours, sterilization time when small using 5 when 100 DEG C), stand and 600 mesh sieves crossed after cooling, receive Collect liquid phase, as high sugar wine base.

The high sugar wine base that ' CSR6R6-888 ' is obtained is named as ' CSR6R6-888 ' high sugar wine base.

The high sugar wine base that ' CSR6R6-666 ' is obtained is named as ' CSR6R6-666 ' high sugar wine base.

The high sugar wine base that ' CSR6R6-777 ' is obtained is named as ' CSR6R6-777 ' high sugar wine base.

3rd, the preparation of low high sugar wine base

The low wine base of the high sugar wine base of 1 parts by volume ' CSR6R6-888 ' and 6 parts by volume ' CSR6R6-888 ' is mixed, obtained ' CSR6R6-888 ' low high sugar wine base.The alcoholic strength of ' CSR6R6-888 ' low high sugar wine base that the present embodiment is obtained is 10%.

The low wine base of the high sugar wine base of 1 parts by volume ' CSR6R6-666 ' and 6 parts by volume ' CSR6R6-666 ' is mixed, obtained ' CSR6R6-666 ' low high sugar wine base.The alcoholic strength of ' CSR6R6-666 ' low high sugar wine base that the present embodiment is obtained is 10%.

The low wine base of the high sugar wine base of 1 parts by volume ' CSR6R6-777 ' and 6 parts by volume ' CSR6R6-777 ' is mixed, obtained ' CSR6R6-777 ' low high sugar wine base.The alcoholic strength of ' CSR6R6-777 ' low high sugar wine base that the present embodiment is obtained is 10%.

The preparation of the high flavonoids applejack of the strong type of embodiment 5, sweet tea

1st, the low high sugar wine base of 1 parts by volume height wine base and 0.5 parts by volume is mixed, first 30 DEG C stand 24 hours, then 0 DEG C 48 hours are stood, 600 mesh sieves are then crossed, liquid phase is collected, is first wine., can first 25 to 45 DEG C of standings 24 to 72 in practical application Hour, then 0 to 4 DEG C stand 24 to 72 hours.

2nd, the first wine that step 1 is obtained is taken, carrying out high and low temperature alternative curing, (first 30 DEG C stand 24 hours, again -18 DEG C standings 48 Hour, repeat 15 times), obtain finished wine., can first 25 to 45 DEG C of standings 24 to 72 hours, -21 DEG C again in practical application 24 to 72 hours are stood to -15 DEG C, is repeated more than 15 times.

' CSR6R6-888 ' prepared by ' CSR6R6-888 ' the height wine base and embodiment 4 prepared using embodiment 3 is low The finished wine that high sugar wine base is prepared is named as the high flavonoids applejack of the strong type of ' CSR6R6-888 ' sweet tea.' CSR6R6-888 ' sweet tea The photo of the strong high flavonoids applejack of type is shown in Fig. 1 and Fig. 2.

' CSR6R6-666 ' prepared by ' CSR6R6-666 ' the height wine base and embodiment 4 prepared using embodiment 3 is low The finished wine that high sugar wine base is prepared is named as the high flavonoids applejack of the strong type of ' CSR6R6-666 ' sweet tea.

' CSR6R6-777 ' prepared by ' CSR6R6-777 ' the height wine base and embodiment 4 prepared using embodiment 3 is low The finished wine that high sugar wine base is prepared is named as the high flavonoids applejack of the strong type of ' CSR6R6-777 ' sweet tea.

Embodiment 6, alcoholic strength detection and sense organ description

Three kinds of high flavonoids applejacks of the strong type of sweet tea prepared by Example 5, detection alcoholic strength and organoleptic indicator.

It the results are shown in Table 2.Alcoholic strength is the 55% high flavonoids applejack of the strong type of sweet tea, and outward appearance is golden yellow, clear, fruit Perfume is fresh and sweet, the wine is fragrantly scented, and entrance is bold, long times of aftertaste.

Table 2

Embodiment 7, the detection of national food safety standard index of correlation

Three kinds of high flavonoids applejacks of the strong type of sweet tea prepared by Example 5, according to national food safety standard GB2757- 2012 and GB2760-2014 detects methanol and content of beary metal.

As a result show, the methanol content in three kinds of high flavonoids applejacks of the strong type of sweet tea prepared by embodiment 5 meets food peace The content of beary metal such as aluminium, manganese in the three kinds of high flavonoids applejacks of the strong type of sweet tea prepared all referring to target national standard, embodiment 5 are equal Meet the national standard of food security index.

Embodiment 8, nutritive and health protection components detection

It is using the Mount Taishan magma white wine that the alcoholic strength that alcoholic strength is produced as 53 degree of Maotai and Taian Shandong brewery is 52 degree Control.The three kinds of high flavonoids applejacks of the strong type of sweet tea and Flavonoid Content, the phenolic acid of control wine that respectively prepared by detection embodiment 5 contain Amount and Mineral Elements Content.

First, extract

1st, 20mL wine to be measured is taken, adjusts pH value to 7 with 1mol/L NaOH, is extracted with ethyl acetate 3 times and (adds 20mL every time Ethyl acetate, is sufficiently mixed rear stratification, then collects organic phase), by three extractions collect it is organic mixing, then 40 DEG C rotary evaporation of solvent, the residue now obtained is flavonoids (neutral phenol), adds 2mL methanol dissolution residual substances, then With 0.22 μm of membrane filtration, HPLC measure is then carried out.

2nd, remaining wine liquid after 3 extractions is taken in step 1, adjusts pH value to 2 with 2mol/L HCl, is extracted with ethyl acetate 3 Secondary (adding 20mL ethyl acetate every time, be sufficiently mixed rear stratification, then collect organic phase), by having that three extractions are collected Machine is mixed, then 40 DEG C of rotary evaporation of solvent, and the residue now obtained is phenolic acid (acidic phenol), adds 2mL methanol molten Residue is solved, then with 0.22 μm of membrane filtration, HPLC measure is then carried out.

2nd, HPLC is detected

1st, HPLC detects the relevant parameter of flavonoids

Highly effective liquid phase chromatographic device model 2695 (Waters, Milford, MA, USA), PDAD (PDA 2998Waters, Milford, MA, USA), season pump and automatic sampler；Splitter is symmetrical C 18 (4.6 × 150 millis Rice, 3.5 μm) post (Waters, Milford, MA, USA), 20 DEG C of column temperature.

Sample size is 10 μ L.

Mobile phase is made up of solvent orange 2 A (containing the aqueous solution of the volume ratio for 2.5% acetic acid) and solvent B (acetonitrile), or, stream Dynamic is mutually solvent B.Elution process (flow velocity 1mL/min)：0-5min, solvent B account for the volume fraction of mobile phase by 3% linear rise To 9%；6-15min, solvent B account for the volume fraction of mobile phase by 9% linear rise to 16%；16-33min, solvent B accounts for flowing The volume fraction of phase is by 16% linear rise to 36.4%；The volume fraction that 34-38min solvents B accounts for mobile phase keeps 100%； Last pillar is repaired 10 minutes (volume fraction that solvent B accounts for mobile phase is 3%).

The Detection wavelength of flavanols and dihydrochalcone is 280nm, and the Detection wavelength of flavonols is 360nm.Flavanols bag Include catechin, epicatechin, procyanidin B 2.Dihydrochalcone includes phloridzin.Flavonols includes Kaempferol, rutin, different Mongolian oak Skin glycosides (Q3Glc), guaijaverin (Quercetin -3-O- α-L- arabopyranoses glycosides), Hyperoside (quercitrin Element -3-O- β-D- galactopyranosides), Quercetin rhamnoside.

The appearance time of catechin standard items is 12.52min.The appearance time of epicatechin standard items is 16.06min. The appearance time of procyanidin B 2 standard items is 13.83min.The appearance time of phloridzin standard items is 27.76min.Kaempferol The appearance time of standard items is 36.86min.The appearance time of rutin standard items is 21.37min.Isoquercitrin standard items go out Peak time is 23.25min.The appearance time of guaijaverin standard items is 24.39min.The appearance time of Hyperoside standard items For 22.64min.The appearance time of Quercetin rhamnoside standard items is 25.35min.Above standard items are purchased from Sigma- Aldrich Co。

2nd, HPLC detects the relevant parameter of phenolic acid

Highly effective liquid phase chromatographic device model 2695 (Waters, Milford, MA, USA), PDAD (PDA 2998Waters, Milford, MA, USA), season pump and automatic sampler；Splitter is symmetrical C 18 (4.6 × 150 millis Rice, 3.5 μm) post (Waters, Milford, MA, USA), 30 DEG C of column temperature.

Sample size is 10 μ L.

Mobile phase is made up of acetonitrile and 0.6% (volume ratio) acetic acid aqueous solution.Elution process (flow velocity 0.5mL/min)：0- 35min, acetonitrile accounts for the volume fraction of mobile phase by 5% linear rise to 35%；36-40min, acetonitrile accounts for the volume integral of mobile phase Number keeps 35%；41-42min, acetonitrile accounts for the volume fraction of mobile phase by 35% linear decline to 5%.

Detection wavelength 280nm.

The appearance time of gallic acid standard items is 6.51min.The appearance time of P-hydroxybenzoic acid standard items is 15.33min.The appearance time of catechuic acid standard items is 15.55min.The appearance time of chlorogenic acid standard items is 15.60min.Coffee The appearance time of coffee acid standard items is 17.29min.The appearance time of syringic acid standard items is 17.3min.Vanillic aldehyde standard items Appearance time is 21.66min.The appearance time of coumaric acid standard items is 22.35min.The appearance time of forulic acid standard items is 24.09min.The appearance time of benzoic acid standard items is 28.35min.The appearance time of cumarin standard items is 30.69min.Meat The appearance time of cinnamic acid standard items is 37.21min.The appearance time of phloretin standard items is 38.49min.Above standard items are equal Purchased from Sigma-Aldrich Co.

3rd, mineral element is detected

Instrument：Inductive coupling Plasma-Mass Spectroscopy (ICP-MS, Thermo Fisher Scientific companies of the U.S.)；WX- 8000 microwave dissolvers (Shanghai EU Apparatus Technology Development Co., Ltd.)；Dura series ultra-pure-water treatment systems；Instrument Running parameter is shown in Table 3.

Table 3

Project	Parameter	Project	Parameter
				Radio-frequency power (W)	1050	Analog voltage (V)	-1900
Atomization gas flow (L/min)	0.86	Pulse voltage (V)	900
				Sample lifting speed (L/min)	1.2	Data acquisition scheme	Jump peak
Ion lens voltage (V)	8.25	Residence time (ms)	50
				Cooling gas flow (L/min)	13.8	Per mass number gathered data point	3
Carrier gas flux (L/min)	0.98	The time of integration (ms)	500

Unit element standard liquid (national non-ferrous metal and electronic material Institute of Analysis, 1000 μ g/mL).Standard The preparation of solution (uses 5%HNO₃The aqueous solution is solvent)：Suction unit element standard liquid, is diluted to 20mg/L storing solution, 200 μ g/L, 400 μ g/L storing solution are diluted to again, then preparation obtains 8 μ g/L, 16 μ g/L, 24 μ g/L, 32 μ g/L, 48 μ successively G/L, 64 μ g/L series standard solution, shake up stand-by.Using 5%HNO₃The aqueous solution marks solution as 0 concentration.Standard is molten Liquid carries out Mass Spectrometer Method, makes standard curve.

Weigh 0.2-1g wine samples to be measured and (be accurate to 0.1mg) into polytetrafluoroethylene (PTFE) counteracting tank, add 5mL concentrated nitric acids Counteracting tank, is then put into microwave dissolver by (concentrated nitric acid mass fraction is about 68%), clears up program and is shown in Table 4.Then it is natural Cooling, after temperature is cooled to below 50 DEG C, takes out counteracting tank and is put into fume hood, open counteracting tank, use ultra-pure water rinse, so The liquid phase in counteracting tank is transferred in 50mL volumetric flasks afterwards, scale is settled to ultra-pure water dilution, then carries out Mass Spectrometer Method. Reference standard curve calculates the content of unit element in wine sample to be measured.

Table 4

Step	Temperature (DEG C)	Soaking time (min)
			1	100	3
2	140	3
			3	160	3
4	180	3
			5	190	15

4th, result

Flavonoid component content in three kinds of high flavonoids applejacks of the strong type of sweet tea and control wine is shown in Table 5.Three kinds of strong types of sweet tea Phenolic acid constituent content in high flavonoids applejack and control wine is shown in Table 6.Three kinds of high flavonoids applejacks of the strong type of sweet tea and right 7 are shown in Table according to the Mineral Elements Content in wine.With high flavonoids excellent germplasm CSR6R6-666, CSR6R6-777 and CSR6R6-888 Transparent apple fruit flavonoid component content, the phenolic acid component of the 3 kinds of high flavonoids applejacks of the strong type of sweet tea that are made for raw material contain Amount and Mineral Elements Content are all remarkably higher than the control wine of the similar number of degrees, and wherein CSR6R6-888 transparent apple fruit is raw material Flavonoid component content, phenolic acid constituent content and Mineral Elements Content highest in the high flavonoids applejack of the strong type of sweet tea being made.

The flavonoid component content of table 5

The phenolic acid constituent content of table 6

The Mineral Elements Content of table 7

SEQUENCE LISTING

<110>Shandong Agricultural University

<120>High flavonoids applejack of the strong type of sweet tea and preparation method thereof

<130> GNCYX171213

<160> 2

<170> PatentIn version 3.5

<210> 1

<211> 22

<212> DNA

<213>Artificial sequence

<400> 1

ggtggtcaaa gatgtgtgtt gt 22

<210> 2

<211> 20

<212> DNA

<213>Artificial sequence

<400> 2

tttgcctgct acccacttca 20

Claims

1. a kind of composition for being used to produce applejack, including height wine base and low high sugar wine base；

The ripening fruits of the height wine base, the low wine base and the high sugar wine base using high flavonoids apple is raw material；

(1) crushing and beating；

(2) sterilize；

(3) pectase is added；

(5) distill and collect the distillate between 85-95 DEG C, then distillate is distilled and collected between 85-95 DEG C again Distillate, repeatedly, until obtaining the distillate that alcoholic strength is more than 80%, as height wine base；

(1) crushing and beating；

(2) sterilize；

(3) pectase is added；

(5) liquid phase is collected, is low wine base；

(1) crushing and beating；

(2) sterilize；

(3) pectase is added；

(5) sterilize；

(6) liquid phase, as high sugar wine base are collected.

2. composition as claimed in claim 1, it is characterised in that：The high flavonoids apple is every kilogram of fresh weight ripening fruits Flavonoid Content be more than 5000mg apple germplasm.

3. composition as claimed in claim 1 or 2, it is characterised in that：

In the step (1) of the preparation method of the height wine base, crushing and beating to pulp granularity is below 1mm；

In the step (1) of the preparation method of the low wine base, crushing and beating to pulp granularity is below 1mm；

4. the composition as described in any in claims 1 to 3, it is characterised in that：

In the step (3) of the preparation method of the height wine base, when pulp temperature is 45-55 DEG C, pectase is added；

In the step (3) of the preparation method of the low wine base, when pulp temperature is 45-55 DEG C, pectase is added；

In the step (3) of the preparation method of the high sugar wine base, when pulp temperature is 45-55 DEG C, pectase is added.

5. the composition as described in any in Claims 1-4, it is characterised in that：

In the step (4) of the preparation method of the height wine base, when pulp temperature is 25-35 DEG C, saccharomyces cerevisiae is added；

In the step (4) of the preparation method of the low wine base, when pulp temperature is 25-35 DEG C, saccharomyces cerevisiae is added；

In the step (4) of the preparation method of the high sugar wine base, when pulp temperature is 25-35 DEG C, saccharomyces cerevisiae is added.

6. application of any composition in applejack is prepared in claim 1 to 5.

7. any described composition prepares the preparation method of applejack, including following step in 6 in a kind of application claim 1 Suddenly：

1. 1 parts by volume height wine base and the low high sugar wine base of 0.3-0.7 parts by volume are mixed, first 25~45 DEG C of standings 24~72 are small When, then 0~4 DEG C stand 24~72 hours, collect liquid phase, be just wine；

8. method as claimed in claim 7, it is characterised in that：The step 2. in, high and low temperature alternative curing is：First 25 ~45 DEG C of standings stand 24~72 hours in 24~72 hours, -21 DEG C again~-15 DEG C, repeat more than 15 times.

9. the applejack that the methods described of claim 7 or 8 is prepared.