CN107099422B - High flavonoids applejack of dry low profile and preparation method thereof - Google Patents
High flavonoids applejack of dry low profile and preparation method thereof Download PDFInfo
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- CN107099422B CN107099422B CN201710522666.7A CN201710522666A CN107099422B CN 107099422 B CN107099422 B CN 107099422B CN 201710522666 A CN201710522666 A CN 201710522666A CN 107099422 B CN107099422 B CN 107099422B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/04—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
- C12H6/00—Methods for increasing the alcohol content of fermented solutions or alcoholic beverages
- C12H6/02—Methods for increasing the alcohol content of fermented solutions or alcoholic beverages by distillation
Abstract
The invention discloses a kind of dry high flavonoids applejacks of low profile and preparation method thereof.Height wine base preparation method, using the ripening fruits of high flavonoids apple as raw material, crushing and beating, sterilizing are added pectase, saccharomyces cerevisiae are added and ferments, distill and collect the distillate between 85 95 DEG C, until alcoholic strength is 80% or more.The preparation method of the high flavonoids applejack of dry low profile provided by the invention：1. 1 parts by volume height wine base is mixed with 5.5 6.5 parts by volume minuent wine bases, first 24~72 hours are stood for 25~45 DEG C, then 0~4 DEG C stands 24~72 hours, collects liquid phase, as first wine；2. the first wine for taking step 1. to obtain carries out high and low temperature alternative curing, obtains finished wine.The high flavonoids applejack of dry low profile provided by the invention not only contains the distinctive functional health ingredients of apples such as flavonoids, phenolic acid and mineral element, but also has stronger irritation and white wine mouthfeel, has huge market foreground.
Description
Technical field
The present invention relates to a kind of dry high flavonoids applejacks of low profile 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 ethyl alcohol in wine, unless otherwise specified, usually
Refer to the percent by volume in wine shared by ethyl alcohol at 20 DEG C.Alcoholic strength generally indicates with x%, also use sometimes 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 rose situation in the consumption of China in apparent in recent years.But low fruit
The alcohol content of wine is low, stimulation is small, cannot effectively meet the demand that custom drinks the crowd of white wine.In order to meet custom drink it is white
The demand of wine crowd usually 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 entire China five
In civilized history in thousand, occupy from literary and artistic creation, entertainment to the everyways such as diet culinary art, health care important
Position.Visitor comes from a distant place, and no wine is not enough to expression deep feeling kindness.Fine moment happy festival time, no wine are not enough to show cheerful and light-hearted satisfied.Spring breeze obtains
Meaning, no wine are not enough to the lofty ideal that gives off one's lofty sentiments.Therefore, although low alcohol fruit wine rises situation in the consumption of China in apparent, have
The white wine of certain wine degree will still have prodigious development space in future.
" doctor's food homology eats nutrition, eats health " has become the common recognition of people.Apple contains more human body and is easy to inhale
The free polyphenol or flavonoids received, it is anti-oxidant, prevent cardiovascular and cerebrovascular disease and it is antitumor etc. all have preferable effect,
And phenolic acid (Phenolicacids) is a kind of organic acid containing phenol ring, has arousing brain, raising spirit, enriching yin beauty treatment, skin whitening, clear
It is pyrolyzed effect of fire, " one day apple, doctor is far from me ", considerable country is all using apple as major consumers fruit in the world
Product and recommend energetically.But for a long time, due to the excessive pursuit to yield and appearance, and some merits such as fruit polyphenol exist
The long korneforos of history is gradually eliminated.For this purpose, seminar where Chen Xuesen professor takes the lead in proposing " functional form (high flavonoids) apple
The concept and its breeding strategy of fruit " improve breeding efficiency, expand answering for high flavonoids apple to improve breeding of new variety scheme
With range, four main measures are taken：First, to the property such as Xinjiang red meat apple and apple variety first generation of hybrid fruit total phenol content
On the basis of shape Research on Genetic Variation, the Apple breeding method (patent No. ZL 2,013 1 of " three select two early one to promote " is proposed and implemented
0205419.6), breeding efficiency significantly improves；Second is that utilizing apple varieties such as ' loud, high-pitched sounds ' of genetic background complexity and Xinjiang red meat
Apple (M.sieversiif.niedzwetzkyana) carries out multi-parent strain hybridization and is 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, it is colonized 40,000 plants of hybrid seedling, and declared " a variety of source 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；Third, be in time examination material with the basicly stable offspring's strain of character,
Quality trait evaluation and the research of development mechanism are carried out, and has achieved many impressive progresses；Fourth, according to " removable, small-sized
Change, be intelligent, pure natural, no added " theory, have developed including the high flavonoids apple crushing and beating device of a kind of stroke type, more
Function wine brewing fractionator, multiple-effect condensation bake the high flavonoids applejack complete equipment for processing such as drinking vessel and separation Wine storage device and processing work
Skill.Currently, having created the apple high-efficient breeding technique system that conventional hybridization is organically combined with biotechnology, formulate a collection of new
Kind and excellent germplasm have developed Variety of Apple highly effective matched cultivation and deep process technology system.It is special to authorize and declare invention
Sharp 30 remainders, are bred as new varieties (being) 16；Correlative study paper 120, wherein more than 20 piece of SCI papers are delivered, these researchs
Achievement is totally in the top standard of international similar research.
Invention content
The object of the present invention is to provide a kind of dry high flavonoids applejacks of low profile and preparation method thereof.
Present invention firstly provides a kind of preparation methods of apple processed goods (height wine base), it is characterised in that：
Using the ripening fruits of high flavonoids apple as raw material, including the following steps that carry out successively：
(1) crushing and beating；
(2) it sterilizes；
(3) pectase is added；
(4) saccharomyces cerevisiae is added and ferments, until the alcoholic strength of fermentation system no longer increases；
(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 obtain the distillate that alcoholic strength is 80% or more, as apple processed goods.
In the preparation method of the height wine base, the ripening fruits of the high flavonoids apple is unique raw material.The height
It spends in the preparation method of 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 tap water.The preparation of the height wine base
In method, the ripening fruits of the high flavonoids apple is that the fruit after impurity elimination is thoroughly cleaned with tap water.
In the step (1) of the preparation method of the height wine base, crushing and beating to pulp granularity is 1mm or less.
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, it is added
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, it is added
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 stating the preparation method of 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 be added 1,000,000 u or more 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 be accordingly added 1,000,000 u-150, ten thousand u 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 pectase of 1,000,000 u-150, ten thousand u is accordingly added in the ripening fruits of high flavonoids apple.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 pectase of 1000000 u-150, ten thousand u.In the step (3) of the preparation method of the height wine base, the addition of the pectase
Amount is：2-3g pectases are accordingly added in the ripening fruits of every kilogram high flavonoids apple.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, add
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, it is added
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 is accordingly added 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 is accordingly added 1010cfu
Saccharomyces cerevisiae.During the fermentation, the temperature of fermentation system, control are continued to monitor since being added after saccharomyces cerevisiae 24 hours
Temperature processed is 15-20 DEG C.When described " until the alcoholic strength of fermentation system no longer increases ", the alcoholic strength of fermentation system is 9%-
12%.The saccharomyces cerevisiae concretely high flavonoids saccharomyces cidri No.1.
It is described " until it is 80% or more 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 described " until obtaining the distillate that alcoholic strength is 80% or more " in (5), it is 80% to distillate concretely to obtain alcoholic strength
Object.
The apple processed goods (height wine base) that any description above method is prepared also belongs to protection scope of the present invention.
The present invention also protects application of the apple processed goods (height wine base) in preparing applejack.
The present invention also protects a kind of preparation method of applejack (the high flavonoids applejack of dry low profile), includes the following steps：
1. 1 parts by volume height wine base is mixed with 5.5-6.5 parts by volume minuent wine bases, first 25~45 DEG C of standings 24~72 are small
When, then 0~4 DEG C stand 24~72 hours, collect liquid phase, as just wine；
2. the first wine for taking step 1. to obtain carries out high and low temperature alternative curing, obtains finished wine.
The step 1. in, 1 parts by volume height wine base is mixed with 6 parts by volume minuent wine bases, first 30 DEG C stand 24 hours,
It stands 48 hours for 0 DEG C again, collects liquid phase, as first wine.The step 1. in, " collect liquid phase " concretely " crosses 600 mesh
Sieve collects liquid phase ".
The step 2. in, high and low temperature alternative curing is：First stand 24~72 hours, again -8 DEG C~-2 for 25~45 DEG C
DEG C stand 24~72 hours, repeat 15 times or more.The step 2. in, high and low temperature alternative curing is：First 30 DEG C quiet
It sets 24 hours, again -5 DEG C and stands 48 hours, repeat 15 times.
The applejack (the high flavonoids applejack of dry low profile) that the method is prepared also belongs to protection scope of the present invention.
The number of degrees of the applejack are 15%-21%, concretely 18%.
Using the ripening fruits of high flavonoids apple as raw material, preparation method in turn includes the following steps the minuent wine base：
(1) crushing and beating；
(2) it sterilizes；
(3) pectase is added；
(4) saccharomyces cerevisiae is added and ferments, until the alcoholic strength of fermentation system no longer increases；
(5) liquid phase is collected, as low wine base.
In the preparation method of the minuent wine base, the ripening fruits of the high flavonoids apple is unique raw material.It is described low
It spends in the preparation method of wine base, the ripening fruits of the high flavonoids apple is the fruit after cleaning.The system of the minuent wine base
In Preparation Method, the ripening fruits of the high flavonoids apple is the fruit after being cleaned with tap water.The preparation of the minuent wine base
In method, the ripening fruits of the high flavonoids apple is that the fruit after impurity elimination is thoroughly cleaned with tap water.
In the step (1) of the preparation method of the minuent wine base, crushing and beating to pulp granularity is 1mm or less.
In the step (2) of the preparation method of the minuent 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 minuent 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 minuent wine base, when pulp temperature is 45-55 DEG C, pectin is added
Enzyme.In the step (3) of the preparation method of the minuent wine base, when it is 45-55 DEG C that pulp, which naturally cools to temperature, it is added
Pectase.In the step (3) of the preparation method of the minuent wine base, when pulp temperature is 45-50 DEG C, pectin is added
Enzyme.In the step (3) of the preparation method of the minuent wine base, when it is 45-50 DEG C that pulp, which naturally cools to temperature, it is added
Pectase.In the step (3) of the preparation method of the minuent wine base, when pulp temperature is 45 DEG C, pectase is added.Institute
In the step (3) for stating the preparation method of 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 minuent wine base, the addition of the pectase is：Every kilogram high flavonoids apple
Ripening fruits accordingly be added 1,000,000 u or more pectase.In the step (3) of the preparation method of the minuent wine base, institute
The addition for stating pectase is：The ripening fruits of the high flavonoids apple of every kilogram of fresh weight be accordingly added 1,000,000 u-150, ten thousand u with
On pectase.In the step (3) of the preparation method of the minuent wine base, the addition of the pectase is：Every kilogram
The pectase of 1,000,000 u-150, ten thousand u is accordingly added in the ripening fruits of high flavonoids apple.The preparation method of the minuent 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 pectase of 1000000 u-150, ten thousand u.In the step (3) of the preparation method of the minuent wine base, the addition of the pectase
Amount is：2-3g pectases are accordingly added in the ripening fruits of every kilogram high flavonoids apple.The preparation method of the minuent 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 minuent 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 minuent wine base, when it is 25-35 DEG C that pulp, which naturally cools to temperature, add
Enter saccharomyces cerevisiae.In the step (4) of the preparation method of the minuent wine base, when pulp temperature is 30 DEG C, wine brewing is added
Yeast.In the step (4) of the preparation method of the minuent wine base, when it is 30 DEG C that pulp, which naturally cools to temperature, it is added
Saccharomyces cerevisiae.In the step (4) of the preparation method of the minuent wine base, the addition of the saccharomyces cerevisiae is：Every kilogram
The ripening fruits of high flavonoids apple is accordingly added 1010Cfu saccharomyces cerevisiaes.The step of the preparation method of the minuent 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 is accordingly added 1010cfu
Saccharomyces cerevisiae.During the fermentation, the temperature of fermentation system, control are continued to monitor since being added after saccharomyces cerevisiae 24 hours
Temperature processed is 15-20 DEG C.When described " until the alcoholic strength of fermentation system no longer increases ", the alcoholic strength of fermentation system is 9%-
12%.The saccharomyces cerevisiae concretely high flavonoids saccharomyces cidri No.1.
In the step (5) of the preparation method of the minuent wine base, described " collecting liquid phase " concretely " crosses 600 mesh
Sieve collects liquid phase ".
The high flavonoids apple is the apple kind that the Flavonoid Content of every kilogram of fresh weight ripening fruits is 5000mg or more
Matter.The high flavonoids apple is the apple germplasm that the Flavonoid Content of every kilogram of fresh weight ripening fruits is 7000mg or more.Institute
State the apple germplasm that Flavonoid Content that high flavonoids apple is every kilogram of fresh weight ripening fruits is 10000mg or more.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 are：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 are：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 are：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 No.1, full name are saccharomycete (Saccharomyces sp.) high flavonoids applejack
Yeast No.1 has been 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 efficiently used, developing both has apple
The distinctive functional component of fruit, the smell of fruits is very sweet, and has stronger irritation and white wine mouthfeel, can meet different consumer groups' need
The high flavonoids applejack of series asked, for elongating Apple Industry chain, rich liquor product Market Diversification, improving enterprise product quality
With benefit, meet market and consumption demand is of great significance.The high flavonoids applejack of dry low profile provided by the invention, both contained
The distinctive functional health ingredient of the apples such as flavonoids, phenolic acid and mineral element, and there is stronger irritation and white wine mouthfeel, wine
Perfume is coordinated, entrance alcohol is flat, it is refreshing and, aftertaste is elegant, and the crowd for drinking low alcohol white spirit is suitble to drink, with great application and popularization value
And huge market foreground.
Description of the drawings
Fig. 1 is the photo that ' CSR6R6-888 ' does the high flavonoids applejack of low profile.
Fig. 2 is the photo that ' CSR6R6-888 ' does the high flavonoids applejack of low profile.
Specific implementation mode
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly
What routine biochemistry reagent shop was commercially available.Quantitative test in following embodiment is respectively provided with three repeated experiments, as a result makes even
Mean value.
Refer to the bibliography of ' Fuji apple ' apple and ' loud, high-pitched sound ' apple：Chen Xuesen, it is 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：0.5 parts by volume, 35% concentrated hydrochloric acid and 99.5 parts by volume methanol are mixed
It closes.
Pectase used in embodiment is the pectase purchased from Ningxia jade of the He family Bioisystech Co., Ltd, correlation ginseng
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 items
Under part, it is a pectinase activity unit (1u) that 1min, which is catalyzed hydrolyzed pectin and generates 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, extracts the genomic DNA of apple pulp, using genomic DNA as template, using the primer of F3 and R3 compositions
To carrying out PCR amplification, following standard interpretation genotype is then pressed：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.
One, the acquisition of ' CSR6R6-888 '
Xinjiang red meat apple hybridizes as parent with plain boiled pork cultivation apple varieties such as ' Fuji apples '.According to Mendelian inheritance
Law, Xinjiang red meat apple (R6R1 genotype) hybridize with plain boiled pork cultivation apple varieties such as ' Fuji apple ' (R1R1 genotype),
Progeny population should be red meat phenotype (R6R1 genotype):Plain boiled pork phenotype (R1R1 genotype)=1:1.But in hybridization F1Dai Qun
The single plant of R6R6 genotype is found that in body.
The single plant of one plant of R6R6 genotype is named as ' CSR6R6-888 '.
' CSR6R6-888 ' has following phenotype：The each sections 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 are excellent.
By way of grafting twig or tissue culture, ' CSR6R6-888 ' is expanded numerous.
Two, the preservation of ' CSR6R6-888 '
' 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 are：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.
Three, the preservation of ' CSR6R6-666 '
' CSR6R6-666 ' is the single plant for another plant of R6R6 genotype that the laboratory where inventor selects early period.
' CSR6R6-666 ' has following phenotype：The each sections such as stem, leaf, flower, pericarp and pulp and each stage of development are equal
For aubergine.
' 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 are：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.
Four, the preservation of ' CSR6R6-777 '
' CSR6R6-777 ' is the single plant for another plant of R6R6 genotype that the laboratory where inventor selects early period.
' 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 are：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.
Five, flavonoid component content analysis
' CSR6R6-888 ', ' CSR6R6-666 ' and ' CSR6R6-777 ' is used as plant to be measured respectively.
1, the transparent apple on plant to be measured is taken, apple pulp is taken.
2, the pulp for taking step 1 to obtain grinds in liquid nitrogen and obtains powder.
3, the powder that 2g steps 2 obtain is weighed, 0.5% methanol hydrochloride solutions of 5mL are added, 4 DEG C stand extraction 2h, then
8000rpm centrifuges 20min, collects supernatant and residue respectively.
4,0.5% methanol hydrochloride solutions of 5mL are added in the residue for taking step 3 to obtain, and 4 DEG C stand extraction 1h, then
8000rpm centrifuges 20min, collects supernatant.
5, the supernatant that step 3 obtains and the supernatant that step 4 obtains are mixed, obtains mixed liquor.
6, the mixed liquor for taking step 5 to obtain, 37 DEG C of revolvings remove methanol, and residue 2-3ml methanol dissolves, then
8000rpm centrifuges 20min, collects supernatant.
7, the supernatant for taking step 6 to obtain collects filtrate with methanol constant volume to 5ml then with 0.45 μm of membrane filtration.
8, 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 columns (100mm × 2.1mm), filler grain
1.7 μm of diameter；45 DEG C of column temperature；1 μ L of sampling volume；
Mobile phase is the mixed liquor of A liquid and B liquid, flow velocity 0.3mL/min；A liquid is acetonitrile, and B liquid is containing 0.2% (volume
Score) formic acid 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 acquisition of electro-spray ionization cation
Pattern (ESI+)；Scanning range 100-1500m/z；Capillary voltage 3.5kV, orifice potential 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 substances 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 the red meat apple first generation of hybrid to bibliography
[J] Scientia Agricultura Sinicas, 2014,47 (11) are evaluated in variation and the excellent strain of functional form apple:2193-2204..
Table 1
The above result shows that ' CSR6R6-888 ', ' CSR6R6-666 ' and ' CSR6R6-777 ' all has very strong class Huang
Ketone synthesis capability, pulp is rich in flavonoids, and contains special flavonols component, is the excellent of high flavonoids apple variety
Germplasm.' CSR6R6-888 ' is better than ' CSR6R6-666 ', and ' CSR6R6-666 ' is better than ' CSR6R6-777 '.
Six, Flavonoid Content measures
' CSR6R6-888 ', ' CSR6R6-666 ' and ' CSR6R6-777 ' is used as plant to be measured respectively.
(1) the transparent apple fruit on plant to be measured is taken, apple pulp is taken.
(2) take 1g pulp, liquid nitrogen grinding that it is water-soluble that 4 DEG C of 65% (volumn concentration) ethyl alcohol being pre-chilled of 10ml are then added
Liquid and mixing, 4 DEG C are protected from light standing extraction 4h, and then 12000g centrifuges 20min, collects supernatant.
(3) test tube is taken, the supernatant that 0.5ml steps (2) obtain is added, then sequentially adds 1mL 5g/100ml NaNO2
Aqueous solution, 1ml 10g/100ml AL (NO3)3Aqueous solution, 4mL 2M NaOH aqueous solutions stand 15min, 8000rpm after mixing
10min is centrifuged, supernatant is taken, then measures light absorption value at 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.
The acquisition of embodiment 2, yeast strain
One, the domestication and separation of unartificial yeast
When 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 detached using plate streaking partition method, each fermentation liquid makees 3 tablets, trained in 28 DEG C of insulating boxs
Support 3d.Morphologic observation is carried out to bacterium colony, picking detaches the good, single bacterium colony with typicalness from tablet, is inoculated in examination respectively
In pipe inclined-plane and sterilized Boiling tube cider, 3 single bacterium colonies are only selected on 3 tablets of each fermentation liquid, and number
Label, 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 set
It is stored in refrigerator.
Two, yeast screening assay
1, Du Shi pipes fermentation screening.
Using Du Shi pipe fermentation methods, under identical condition of culture, measures the speed of yeast strain aerogenesis bubble and providing
In time aerogenesis steep number, tentatively more each saccharomycete rise ferment ability and fermentability, it is excellent to filter out fermenting property
Yeast strain.Test parallel be repeated 3 times.Bacterial strain activation condition：25 DEG C constant temperature incubation is for 24 hours in the cider of 10 ° of Brix.Hair
Ferment condition：20 DEG C of static fermentation 48h in the cider of 15 ° of Brix.
2, saccharomycetes to make fermentation power tests (CO2Weight-loss method).
3, saccharomycete coherency compares (yeast number comparison method).
4, the resistance to ethyl alcohol 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, more each yeast strain is to ethyl alcohol, SO2Tolerance degree, further determine that suitable
The bacterial strain of apple liquor brewing.Test parallel be repeated 3 times.Bacterial strain activation condition：25 DEG C of constant temperature incubations in the cider of 10 ° of Brix
24h.Fermentation condition：20 DEG C in the cider of 15 ° of Brix static fermentation 96h (due to ethyl alcohol or SO2Presence, to saccharomycete
Fermentation will produce inhibiting effect to varying degrees, therefore aerogenesis observing time is extended for 96h).Inoculum concentration：(6~8) × 107
A/ml.
5, saccharomycete brews cider fermentation experiment.
500ml triangular flasks are taken, 400ml ciders (20 ° of Brix of pol) are added, bacteria concentration is accessed by the inoculum concentration of 20ml/L
For (3~3.6) × 106The saccharomycete seed liquor of a/ml, 20 DEG C of fermentations.Fermentation carries out tank switching to 15d, removes wine foot, then old
After making 10d, measures the physical and chemical index of applejack and carry out organoleptic analysis.
Compared by yeast fermenting power, cohesiveness compares, resistance to SO2, resistance to ethyl alcohol ability compare and ferment to it gained apple
The physical and chemical index of wine is analyzed and organoleptic quality evaluations, filters out three plants of best applejack saccharomyces cerevisiaes, and it is yellow to be respectively designated as high class
Ketone saccharomyces cidri No.1, high flavonoids saccharomyces cidri two, high flavonoids saccharomyces cidri three.
Three, the performance for the three saccharomycetes production applejack that comparison step two obtains
Yeast to be measured is respectively：High flavonoids saccharomyces cidri No.1, high flavonoids saccharomyces cidri two or high class are yellow
Ketone saccharomyces cidri three.
1, it takes ' CSR6R6-888 ' without rotten ripening fruits, is thoroughly cleaned with tap water.
2, after completing step 1, fruit is taken, carries out crushing and beating, until pulp granularity is 1mm or less.
3, after completing step 2, pulp is sterilized (80 DEG C, 12 hours).
4, after completing step 3, pectase is added when pulp temperature is 50 DEG C (per the fruit of 1000g fresh weights for natural cooling
It is real, 2-3g pectases are added).
5, after completing step 4, continue natural cooling, when pulp temperature is 30 DEG C, it is (fresh per 1000g that yeast to be measured is added
The fruit of weight is added 1010The yeast to be measured of cfu), it ferments.The alcoholic strength that fermentation system is continued to monitor in fermentation process, when
Stop fermentation when alcoholic strength no longer increases (alcoholic strength of system is 9%-12% at this time).In fermentation process, from addition ferment to be measured
Start to continue to monitor the temperature of fermentation system after 24 hours female, controlled at 15-20 DEG C.
Physical and chemical index analysis and organoleptic quality evaluations are carried out to obtained applejack, high flavonoids saccharomyces cidri No.1
Indices are most ideal, are resistant to 12% ethyl alcohol, for the former wine alcoholic strength of fermentation up to 11.9%, wine body clear has apple
The typical flavor of fruit wine.
Four, the identification of high flavonoids saccharomyces cidri No.1
Morphological feature：In oval spherical, the vacuole for having apparent nucleus and differing 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 cannot 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.
Five, the preservation of high flavonoids saccharomyces cidri No.1
High flavonoids saccharomyces cidri No.1, full name are saccharomycete (Saccharomyces sp.) high flavonoids applejack
Yeast No.1 has been 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.
Embodiment 3 and embodiment 4 are carried out as saccharomyces cerevisiae using high flavonoids saccharomyces cidri No.1.
The preparation of embodiment 3, height wine base
The apple on ' CSR6R6-888 ', the apple on ' CSR6R6-666 ' and the apple on ' CSR6R6-777 ' is respectively adopted
Fruits prepare height wine base.
1, it takes without rotten transparent apple fruit, impurity elimination is thoroughly cleaned with tap water.
2, after completing step 1, fruit is taken, carries out crushing and beating, until pulp granularity is 1mm or less.
3, after completing step 2, pulp is sterilized (100 DEG C, 5 hours；80-100 DEG C of sterilizing can be used in practical application
5-12 hours, longer sterilization time was used when the temperature is low, when temperature is higher using shorter sterilization time, such as when
Using 10-12 hours sterilization times at 80 DEG C, sterilization time when small using 5 when 100 DEG C).
4, after completing step 3, natural cooling (in practical application, can be used 45-55 DEG C, specifically when pulp temperature is 45 DEG C
Can be used 45-50 DEG C) when, pectase is added and (per the fruit of 1000g fresh weights, 2-3g pectases are added；The enzyme activity of 2-3g pectases
For 1,000,000 u-150, ten thousand u or more).
5, after completing step 4, continue natural cooling, when pulp temperature is 30 DEG C (in practical application, can be used 25-35 DEG C)
When, saccharomyces cerevisiae is added and (per the fruit of 1000g fresh weights, is added 1010Cfu saccharomyces cerevisiaes), it ferments.Continue in fermentation process
The alcoholic strength for monitoring fermentation system stops fermentation when alcoholic strength no longer increases (alcoholic strength of system is 9%-12% at this time).
In fermentation process, the temperature of fermentation system is continued to monitor since being added after saccharomyces cerevisiae 24 hours, controlled at 15-20
℃。
6, it after completing step 5, is distilled, collects the distillate between 85-95 DEG C.The alcoholic strength of the distillate is about
20%.
7, the distillate for taking step 6 to obtain, is repeatedly distilled, and collects the distillate between 85-95 DEG C every time,
Until obtaining alcoholic strength as 80% or more distillate, 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 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 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 obtained
The alcoholic strength of ' CSR6R6-777 ' height wine base is 80%.
The preparation of embodiment 4, low wine base
The apple on ' CSR6R6-888 ', the apple on ' CSR6R6-666 ' and the apple on ' CSR6R6-777 ' is respectively adopted
Fruits prepare low wine base.
1, it takes without rotten transparent apple fruit, impurity elimination is thoroughly cleaned with tap water.
2, after completing step 1, fruit is taken, carries out crushing and beating, until pulp granularity is 1mm or less.
3, after completing step 2, pulp is sterilized (100 DEG C, 5 hours；80-100 DEG C of sterilizing can be used in practical application
5-12 hours, longer sterilization time was used when the temperature is low, when temperature is higher using shorter sterilization time, such as when
Using 10-12 hours sterilization times at 80 DEG C, sterilization time when small using 5 when 100 DEG C).
4, after completing step 3, natural cooling (in practical application, can be used 45-55 DEG C, specifically when pulp temperature is 45 DEG C
Can be used 45-50 DEG C) when, pectase is added and (per the fruit of 1000g fresh weights, 2-3g pectases are added；The enzyme activity of 2-3g pectases
For 1,000,000 u-150, ten thousand u or more).
5, after completing step 4, continue natural cooling, when pulp temperature is 30 DEG C (in practical application, can be used 25-35 DEG C)
When, saccharomyces cerevisiae is added and (per the fruit of 1000g fresh weights, is added 1010Cfu saccharomyces cerevisiaes), it ferments.Continue in fermentation process
The alcoholic strength for monitoring fermentation system stops fermentation when alcoholic strength no longer increases (alcoholic strength of system is 9%-12% at this time).
In fermentation process, the temperature of fermentation system is continued to monitor since being added after saccharomyces cerevisiae 24 hours, controlled at 15-20
℃。
6, after completing step 5,600 mesh sieve is crossed, liquid phase is collected, as low wine base.
The low wine base that ' CSR6R6-888 ' is obtained is named as ' CSR6R6-888 ' low wine base.What the present embodiment obtained
The alcoholic strength of ' CSR6R6-888 ' minuent wine base is 10%.
The low wine base that ' CSR6R6-666 ' is obtained is named as ' CSR6R6-666 ' low wine base.What the present embodiment obtained
The alcoholic strength of ' CSR6R6-666 ' minuent wine base is 10%.
The low wine base that ' CSR6R6-777 ' is obtained is named as ' CSR6R6-777 ' low wine base.What the present embodiment obtained
The alcoholic strength of ' CSR6R6-777 ' minuent wine base is 10%.
The preparation of embodiment 5, the high flavonoids applejack of dry low profile
1,1 parts by volume height wine base is mixed with 6 parts by volume minuent wine bases, first stands 24 hours for 30 DEG C, then 0 DEG C stands 48
Hour, 600 mesh sieve is then crossed, liquid phase is collected, as first wine.In practical application, can it is first 25 to 45 DEG C stand 24 to 72 hours, then
0 to 4 DEG C stands 24 to 72 hours.
2, the first wine that step 1 obtains is taken, high and low temperature alternative curing is carried out and (first stands 24 hours, again -5 DEG C standings 48 for 30 DEG C
Hour, repeat 15 times), obtain finished wine.In practical application, can it is first 25 to 45 DEG C stand 24 to 72 hours, again -8 DEG C to -
2 DEG C stand 24 to 72 hours, repeat 15 times or more.
' CSR6R6-888 ' prepared by ' CSR6R6-888 ' the height wine base and embodiment 4 prepared using embodiment 3 is low
The finished wine that wine base is prepared is named as ' CSR6R6-888 ' and does the high flavonoids applejack of low profile.' CSR6R6-888 ' does low profile
The photo of high flavonoids applejack 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 wine base is prepared is named as ' CSR6R6-666 ' and does the high flavonoids applejack of low profile.
' CSR6R6-777 ' prepared by ' CSR6R6-777 ' the height wine base and embodiment 4 prepared using embodiment 3 is low
The finished wine that wine base is prepared is named as ' CSR6R6-777 ' and does the high flavonoids applejack of low profile.
Embodiment 6, alcoholic strength detection and sense organ description
Three kinds of dry high flavonoids applejacks of low profile prepared by Example 5 detect alcoholic strength and organoleptic indicator.
It the results are shown in Table 2.The high flavonoids applejack of dry low profile that alcoholic strength is 18%, appearance are orange red, clear, wine
Perfume is coordinated, fruity protrudes, entrance alcohol is flat, it is refreshing and, aftertaste is elegant.
Table 2
Embodiment 7, the detection of national food safety standard index of correlation
Three kinds of dry high flavonoids applejacks of low profile prepared by Example 5, according to national food safety standard GB2757-
2012 and GB2760-2014 detects methanol and content of beary metal.
The result shows that the methanol content in three kinds of dry high flavonoids applejacks of low profile prepared by embodiment 5 meets food peace
All referring to target national standard, the content of beary metal such as aluminium, manganese in three kinds of dry high flavonoids applejacks of low profile prepared by embodiment 5 are equal
Meet the national standard of food security index.
Embodiment 8, nutritive and health protection components detection
Abundant dry white wine and the wine that alcoholic strength with Zhangyu Wine Making Co., Ltd., Yantai City's production is 12%
The abundant claret that precision is 12% is control.Three kinds of dry high flavonoids apples of low profile that respectively prepared by detection embodiment 5
The Flavonoid Content and phenolic content of wine and control wine.
One, it extracts
1,20mL wine to be measured is taken, with 1mol/L NaOH tune pH value to 7,3 times is extracted with ethyl acetate and (20mL is added every time
Ethyl acetate is sufficiently mixed rear stratification, then collects organic phase), the organic of collection will be extracted three times to be mixed, then 40
DEG C rotary evaporation of solvent, the residue obtained at this time is flavonoids (neutral phenol), 2mL methanol dissolution residual substances is added, then
With 0.22 μm of membrane filtration, HPLC measurement is then carried out.
2, remaining wine liquid is extracted with ethyl acetate 3 with 2mol/L HCl tune pH value to 2 after taking in step 13 extractions
Secondary (20mL ethyl acetate being added every time, be sufficiently mixed rear stratification, then collect organic phase), has what extraction three times was collected
Machine mixes, then 40 DEG C of rotary evaporation of solvent, and the residue obtained at this time is phenolic acid (acidic phenol), and it is molten that 2mL methanol is added
Residue is solved, then with 0.22 μm of membrane filtration, then carries out HPLC measurement.
Two, HPLC is detected
1, HPLC detects the relevant parameter of flavonoids
Highly effective liquid phase chromatographic device model 2695 (Waters, Milford, MA, USA), diode array detector
(PDA 2998 Waters, Milford, MA, USA), season pumps and automatic sampler；Splitter is (4.6 × 150 millis of symmetrical C 18
Rice, 3.5 μm) column (Waters, Milford, MA, USA), 20 DEG C of column temperature.
Sample size is 10 μ L.
Mobile phase is made of solvent A (aqueous solution containing the acetic acid that volume ratio is 2.5%) and solvent B (acetonitrile), alternatively, 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 account 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 packet
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.The above standard items are purchased from Sigma-
Aldrich Co。
2, HPLC detects the relevant parameter of phenolic acid
Highly effective liquid phase chromatographic device model 2695 (Waters, Milford, MA, USA), diode array detector
(PDA 2998 Waters, Milford, MA, USA), season pumps and automatic sampler；Splitter is (4.6 × 150 millis of symmetrical C 18
Rice, 3.5 μm) column (Waters, Milford, MA, USA), 30 DEG C of column temperature.
Sample size is 10 μ L.
Mobile phase is made of acetonitrile and 0.6% (volume ratio) acetic acid aqueous solution.Elution process (flow velocity 0.5mL/min)：0-
35min, acetonitrile account for the volume fraction of mobile phase by 5% linear rise to 35%；36-40min, acetonitrile account for the volume point of mobile phase
Number keeps 35%；41-42min, acetonitrile account 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 ferulic 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.The above standard items are equal
Purchased from Sigma-Aldrich Co.
Three, result
Flavonoid component content in three kinds of dry high flavonoids applejacks of low profile and two kinds of control wine is shown in Table 3.Three kinds dry
Phenolic acid constituent content in the high flavonoids applejack of low profile and two kinds of control wine is shown in Table 4.With high flavonoids excellent germplasm
The transparent apple fruit of CSR6R6-666, CSR6R6-777 and CSR6R6-888 are 3 kinds of dry high flavonoids of low profile made of raw material
The flavonoid component content of applejack is all remarkably higher than the control wine of two kinds of similar number of degrees with phenolic acid constituent content, wherein
The transparent apple fruit of CSR6R6-888 is that flavonoid component content and phenol in the high flavonoids applejack of low profile are done made of raw material
Acid constituents content highest.
3 flavonoid component content of table
4 phenolic acid constituent content of table
SEQUENCE LISTING
<110>Shandong Agricultural University
<120>High flavonoids applejack of dry low profile and preparation method thereof
<130> GNCYX171207
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 22
<212> DNA
<213>Artificial sequence
<400> 1
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<210> 2
<211> 20
<212> DNA
<213>Artificial sequence
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tttgcctgct acccacttca 20
Claims (6)
1. a kind of preparation method of applejack, includes the following steps：
1. 1 parts by volume height wine base is mixed with 5.5-6.5 parts by volume minuent wine bases, first 24~72 hours are stood for 25~45 DEG C,
It stands 24~72 hours for 0~4 DEG C again, collects liquid phase, as first wine；
2. the first wine for taking step 1. to obtain carries out high and low temperature alternative curing, obtains finished wine；
Using the ripening fruits of high flavonoids apple as unique raw material, preparation method in turn includes the following steps the height wine base：
(1) crushing and beating；
(2) it sterilizes；
(3) pectase is added；
(4) saccharomyces cerevisiae is added and ferments, until the alcoholic strength of fermentation system no longer increases；
(5) it distills and collects the distillate between 85-95 DEG C, then distill distillate again and collect between 85-95 DEG C
Distillate, repeatedly, until obtaining alcoholic strength as 80% or more distillate, as height wine base；
Using the ripening fruits of high flavonoids apple as unique raw material, preparation method in turn includes the following steps the minuent wine base：
(1) crushing and beating；
(2) it sterilizes；
(3) pectase is added；
(4) saccharomyces cerevisiae is added and ferments, until the alcoholic strength of fermentation system no longer increases；
(5) liquid phase is collected, as low wine base；
In the height wine base and the low wine base, the high flavonoids apple as unique raw material is ' CSR6R6-888 '；
The full name of ' CSR6R6-888 ' is apple (Malus domestica) CSR6R6-888, preservation registration number CGMCC
NO.14295；
In the preparation method of the height wine base and the low wine base, the saccharomyces cerevisiae is high flavonoids saccharomyces cidri
No.1；The full name of high flavonoids saccharomyces cidri No.1 is saccharomycete (Saccharomyces sp.) high flavonoids applejack ferment
Female No.1, preservation registration number are CGMCC NO.14250.
2. the method as described in claim 1, it is characterised in that：In the step (1), crushing and beating to pulp granularity is 1mm
Below.
3. the method as described in claim 1, it is characterised in that：In the step (3), when pulp temperature is 45-55 DEG C, add
Enter pectase.
4. the method as described in claim 1, it is characterised in that：In the step (4), when pulp temperature is 25-35 DEG C, add
Enter saccharomyces cerevisiae.
5. the method as described in claim 1, 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, -8 DEG C again~-2 DEG C, repeat 15 times or more.
6. the applejack that any the method is prepared in claim 1 to 5.
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| CN101531960A (en) * | 2009-01-24 | 2009-09-16 | 广东岭南为多生命高科有限公司 | Wine-making preparation method adopting litchi fermentation |
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| CN103146536A (en) * | 2013-04-02 | 2013-06-12 | 罗福国 | Method for brewing bamboo health-care wine by high concentration bamboo juice |
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| CN106613921A (en) * | 2017-02-21 | 2017-05-10 | 山东农业大学 | Method for continuously improving red characters of apples by combining continuous multi-generation bud mutation selection and cross breeding |
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| 苹果酿造工艺研究;王思勰;《中国优秀硕士学位论文全文数据库工程科技Ι辑》;20160131(第1期);第4,24-27,53-58页 * |
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