CN107118900B - Do soft high flavonoids applejack of type and preparation method thereof - Google Patents

Abstract

The invention discloses a dry soft high-flavonoid cider and a preparation method thereof. The preparation method of the low-alcohol alcohol base comprises the following steps: using ripe fruits of high-flavonoid apples as raw materials, crushing, beating, and sterilizing, adding pectinase and brewing yeast for fermentation, and collecting the liquid phase. The preparation method of the dry soft high-flavonoid cider provided by the present invention comprises the following steps: ① Mix 1 volume part of high alcohol base with 1.8-2.2 volume parts of low alcohol base, and first stand at 25-45°C for 24-72 hour, then stand still at 0-4°C for 24-72 hours, collect the liquid phase, which is the first wine; ②take the first wine obtained in step ①, and carry out high and low temperature alternating aging to obtain the finished wine. The dry and soft high-flavonoid cider provided by the invention not only contains flavonoids, phenolic acids and mineral elements and other apple-specific functional health-care ingredients, but also has strong irritation and taste of liquor, and has a huge market prospect.

Description

Dry soft high-flavonoid cider and preparation method thereof

technical field

The invention relates to a dry soft high-flavonoid cider and a preparation method thereof.

Background technique

The degree of wine, also known as alcohol content, refers to the volume percentage of ethanol in wine. If there is no special instruction, it usually refers to the volume percentage of ethanol in wine at 20°C. Alcohol is generally expressed by x%, sometimes also expressed by x% (V/V) or x%vol or x°. An alcohol content of x% means that at 20°C, 100 unit volumes of alcohol contain x unit volumes of ethanol.

Low-alcohol fruit wines, such as dry red wine and dry white wine, contain flavonoids (anthocyanins), essential amino acids and mineral elements and other nutritional and health ingredients. The consumption in China has shown a significant upward trend in recent years. However, low-alcohol fruit wine has low alcohol content and little stimulating effect, and cannot effectively meet the needs of people who are used to drinking liquor. In order to meet the needs of people who are accustomed to drinking liquor, in the prior art, the alcohol content of low-alcohol fruit wine is usually increased by adding sugar or edible alcohol to the fermented liquid.

Wine and wine culture are an important part of Chinese diet and social culture. Liquor has permeated the entire 5,000-year history of civilization in China, and plays an important role in various aspects from literary and artistic creation, cultural entertainment to food and cooking, and health care. When guests come from afar, no wine is not enough to express their deep affection. On good days and festivals, no wine is not enough to show joy and contentment. The spring breeze is proud, and no wine is not enough to express pride and ambition. Therefore, although the consumption of low-alcohol fruit wine in China is on the rise, there is still a lot of room for development of liquor with a certain alcohol content in the future.

"Medicine and food come from the same source, eat nutritiously, eat healthily" has become the consensus of people. Apple fruit contains more free polyphenols or flavonoids that are easily absorbed by the human body, which have good effects in anti-oxidation, prevention of cardiovascular and cerebrovascular diseases, and anti-tumor. The organic acid has the effects of refreshing the brain, nourishing yin and beautifying the skin, whitening the skin, clearing away heat and relieving fire, "an apple a day keeps the doctor away", quite a few countries in the world strongly recommend apples as the main consumer fruit. But for a long time, due to the excessive pursuit of yield and appearance, some excellent traits such as fruit polyphenols have been gradually eliminated in the long river of history. For this reason, Professor Chen Xuesen's research group took the lead in proposing the concept of "functional (high flavonoid) apple" and its breeding ideas. Four measures were proposed: first, based on the research on genetic variation of traits such as total phenolic content in fruits of Xinjiang red-flesh apples and apple varieties hybrids, the apple breeding method of "three selections, two early ones and one promotion" was proposed and implemented (Patent No. ZL 2013 10205419.6), the breeding efficiency has been significantly improved; the second is to use apple varieties such as 'Gala' with complex genetic background and Xinjiang red flesh apple (M. At present, 40 first- and second-generation segregation populations of hybrid (back) crosses have been constructed, 40,000 hybrid seedlings have been planted, and the "Multiple Source Quality Breeding Method for Fruit Trees" (patent application number 2015 10428448.8) and "Easy Coloring Apple Variety Cultivation Method" (patent application number 201510890141.X) and two breeding technology invention patents; the third is to use the offspring lines with basically stable traits as test materials in a timely manner to conduct quality trait evaluation and development mechanism research, and has obtained many Important progress; Fourth, according to the concept of "removable, miniaturized, intelligent, pure natural, no additives", it has developed a stroke-type high-flavonoid apple crushing beater, a multi-functional brewing fractionator, and a multi-effect condensation roaster. Complete sets of equipment and processing technology for high flavonoid cider processing such as wine vessels and separated wine storage vessels. At present, a high-efficiency apple breeding technology system that combines conventional hybridization and biotechnology has been established, a number of new varieties and excellent germplasm have been created, and a high-efficiency cultivation and deep processing technology system for new apple varieties has been developed. More than 30 invention patents have been authorized and declared, and 16 new varieties (series) have been bred; 120 related research papers have been published, including more than 20 SCI papers. These research results are generally at the leading level of similar international research.

Contents of the invention

The object of the present invention is to provide a kind of dry and soft high-flavonoid cider and its preparation method.

The present invention at first provides a kind of preparation method of apple processed product (low alcohol base), is characterized in that:

The ripe fruit of high-flavonoid apples is used as raw material, and the following steps are carried out in sequence:

(1) crushing and beating;

(2) Sterilization;

(3) adding pectinase;

(4) adding Saccharomyces cerevisiae and fermenting until the alcohol content of the fermentation system no longer increases;

(5) Collect the liquid phase, which is apple processed product.

In the preparation method of the low alcohol base, the ripe fruit of the high-flavonoid apple is the only raw material. In the preparation method of the low alcohol base, the ripe fruit of the high-flavonoid apple is the fruit after washing. In the preparation method of the low alcohol base, the ripe fruit of the high-flavonoid apple is the fruit washed with tap water. In the preparation method of the low-alcoholic alcohol base, the ripe fruit of the high-flavonoid apple is the fruit that has been thoroughly washed and removed with tap water.

In the step (1) of the preparation method of the low alcohol base, crushing and beating until the pulp particle size is below 1mm.

In the step (2) of the preparation method of the low alcohol base, the sterilization condition is: 80-100° C. for 5-12 hours. In the step (2) of the preparation method of the low-alcohol base, the sterilization conditions may specifically be: 100° C., 5 hours.

In the step (3) of the preparation method of the low alcohol base, when the temperature of the pulp is 45-55° C., pectinase is added. In the step (3) of the preparation method of the low alcohol base, when the pulp is naturally cooled to a temperature of 45-55° C., pectinase is added. In the step (3) of the preparation method of the low alcohol base, when the temperature of the pulp is 45-50° C., pectinase is added. In the step (3) of the preparation method of the low alcohol base, when the pulp is naturally cooled to a temperature of 45-50° C., pectinase is added. In the step (3) of the preparation method of the low alcohol base, when the temperature of the pulp is 45° C., pectinase is added. In the step (3) of the preparation method of the low alcohol base, when the pulp is naturally cooled to a temperature of 45° C., pectinase is added. In the step (3) of the preparation method of the low-alcoholic alcohol base, the amount of the pectinase added is: correspondingly add more than 1 million u of the pectinase per kilogram of ripe fruit of high-flavonoid apples. In the step (3) of the preparation method of the low alcohol base, the addition amount of the pectinase is: more than 1 million u-1.5 million u are correspondingly added per kilogram of fresh weight of ripe fruit of high flavonoid apples pectinase. In the step (3) of the preparation method of the low-alcohol alcohol base, the amount of the pectinase added is: 1 million u-1.5 million u of pectinase is added to each kilogram of mature fruit of high-flavonoid apples . In the step (3) of the preparation method of the low alcohol base, the addition amount of the pectinase is: 1 million u-1.5 million u of pectinase is added correspondingly to the ripe fruit of high flavonoid apples per kilogram of fresh weight pectinase. In the step (3) of the preparation method of the low-alcoholic alcohol base, the addition amount of the pectinase is: 2-3g pectinase is correspondingly added per kilogram of ripe fruit of high-flavonoid apples. In the step (3) of the preparation method of the low-alcoholic alcohol base, the addition amount of the pectinase is: 2-3g pectinase is correspondingly added per kilogram of ripe fruit of high-flavonoid apples with fresh weight.

In the step (4) of the preparation method of the low alcohol base, when the temperature of the pulp is 25-35° C., add Saccharomyces cerevisiae. In the step (4) of the preparation method of the low alcohol base, when the pulp is naturally cooled to a temperature of 25-35° C., Saccharomyces cerevisiae is added. In the step (4) of the preparation method of the low alcohol base, when the temperature of the pulp is 30° C., Saccharomyces cerevisiae is added. In the step (4) of the preparation method of the low alcohol base, when the pulp is naturally cooled to a temperature of 30° C., Saccharomyces cerevisiae is added. In the step (4) of the preparation method of the low-alcohol base, the addition amount of the Saccharomyces cerevisiae is: 10 ¹⁰ cfu Saccharomyces cerevisiae is added correspondingly to every kilogram of ripe fruit of high-flavonoid apples. In the step (4) of the preparation method of the low-alcoholic alcohol base, the addition amount of the Saccharomyces cerevisiae is: 10 ¹⁰ cfu Saccharomyces cerevisiae is correspondingly added per kilogram of fresh weight of ripe fruit of high-flavonoid apples. During the fermentation process, the temperature of the fermentation system was continuously monitored 24 hours after adding Saccharomyces cerevisiae, and the temperature was controlled at 15-20°C. When said "until the alcohol content of the fermentation system no longer increases", the alcohol content of the fermentation system is 9%-12%. The Saccharomyces cerevisiae can specifically be high flavonoid cider yeast No. 1.

In the step (5) of the preparation method of the low alcohol base, the "collecting the liquid phase" may specifically be "passing through a 600-mesh sieve to collect the liquid phase".

Apple processed products (low alcohol base) prepared by any of the above methods also belong to the protection scope of the present invention.

The invention also protects the application of the apple processed product (low alcohol base) in the preparation of cider.

The present invention also protects a preparation method of cider (dry soft high-flavonoid cider), comprising the following steps:

①Mix 1 volume part of high-alcoholic alcohol base with 1.8-2.2 volume parts of low-alcohol alcohol base, first stand at 25-45°C for 24-72 hours, then at 0-4°C for 24-72 hours, and collect the liquid phase, which is First wine;

② Take the primary wine obtained in step ①, and carry out high and low temperature alternate aging to obtain the finished wine.

In the step ①, mix 1 volume part of high-alcohol base and 2 volume parts of low-alcohol base, firstly let it stand at 30°C for 24 hours, then let it stand at 0°C for 48 hours, and collect the liquid phase, which is the initial liquor. In the step ①, the "collecting the liquid phase" can specifically be "passing through a 600-mesh sieve to collect the liquid phase".

In the step ②, the alternating high and low temperature curing is as follows: first stand at 25-45°C for 24-72 hours, then at -13°C--7°C for 24-72 hours, repeat more than 15 times. In the step ②, the alternating high and low temperature curing is as follows: firstly standing at 30°C for 24 hours, then standing at -10°C for 48 hours, repeating 15 times.

The cider (dry soft high-flavonoid cider) prepared by the method also belongs to the protection scope of the present invention. The alcohol content of the cider is 30%-36%, specifically 33%.

The high-alcoholic alcohol base uses ripe fruit of high-flavonoid apples as raw materials, and the preparation method includes the following steps in sequence:

(1) crushing and beating;

(2) Sterilization;

(3) adding pectinase;

(4) adding Saccharomyces cerevisiae and fermenting until the alcohol content of the fermentation system no longer increases;

(5) Distill and collect the distillate between 85-95°C, then distill the distillate again and collect the distillate between 85-95°C, several times until the distillate with an alcohol content of 80% or more is obtained The product is a high alcohol base.

In the highly alcohol-based preparation method, the ripe fruit of the high-flavonoid apple is the only raw material. In the preparation method of the high alcohol base, the ripe fruit of the high flavonoid apple is the fruit after washing. In the preparation method with high alcohol base, the ripe fruit of the high-flavonoid apple is the fruit after washing with tap water. In the preparation method with high alcohol base, the ripe fruit of the high-flavonoid apple is the fruit after thoroughly washing and removing impurities with tap water.

In the step (1) of the high-alcohol-based preparation method, crushing and beating until the pulp particle size is below 1mm.

In the step (2) of the highly alcohol-based preparation method, the sterilization conditions are: 80-100°C for 5-12 hours. In the step (2) of the highly alcohol-based preparation method, the sterilization conditions may specifically be: 100° C., 5 hours.

In the step (3) of the highly alcohol-based preparation method, when the temperature of the pulp is 45-55° C., pectinase is added. In the step (3) of the highly alcohol-based preparation method, when the fruit pulp is naturally cooled to a temperature of 45-55° C., pectinase is added. In the step (3) of the highly alcohol-based preparation method, when the temperature of the pulp is 45-50° C., pectinase is added. In the step (3) of the highly alcohol-based preparation method, when the fruit pulp is naturally cooled to a temperature of 45-50° C., pectinase is added. In the step (3) of the highly alcohol-based preparation method, when the temperature of the pulp is 45° C., pectinase is added. In the step (3) of the highly alcohol-based preparation method, when the fruit pulp is naturally cooled to a temperature of 45° C., pectinase is added. In the step (3) of the high-alcohol-based preparation method, the amount of pectinase added is: correspondingly add more than 1 million u of pectinase per kilogram of ripe fruit of high-flavonoid apples. In the step (3) of the preparation method of the high alcohol base, the addition amount of the pectinase is as follows: 1 million u-1.5 million u of more than 1 million u-1.5 million u of the mature fruit of the high-flavonoid apple per kilogram of fresh weight is correspondingly added pectinase. In the step (3) of the high-alcohol-based preparation method, the amount of pectinase added is: 1 million u-1.5 million u of pectinase is correspondingly added per kilogram of mature fruit of high-flavonoid apples. In the step (3) of the preparation method of the high alcohol base, the addition amount of the pectinase is as follows: 1 million u-1.5 million u of the ripe fruit of high flavonoid apples per kilogram of fresh weight is correspondingly added. Glue enzyme. In the step (3) of the highly alcohol-based preparation method, the added amount of the pectinase is: 2-3 g of pectinase is correspondingly added per kilogram of ripe fruit of high-flavonoid apples. In the step (3) of the highly alcohol-based preparation method, the added amount of the pectinase is: 2-3g pectinase is correspondingly added per kilogram of fresh weight of ripe fruit of high-flavonoid apples.

In the step (4) of the highly alcohol-based preparation method, when the temperature of the pulp is 25-35° C., add Saccharomyces cerevisiae. In the step (4) of the highly alcohol-based preparation method, when the pulp is naturally cooled to a temperature of 25-35° C., Saccharomyces cerevisiae is added. In the step (4) of the highly alcohol-based preparation method, when the temperature of the pulp is 30° C., Saccharomyces cerevisiae is added. In the step (4) of the highly alcohol-based preparation method, when the pulp is naturally cooled to a temperature of 30° C., Saccharomyces cerevisiae is added. In the step (4) of the highly alcohol-based preparation method, the amount of Saccharomyces cerevisiae added is: 10 ¹⁰ cfu Saccharomyces cerevisiae is correspondingly added per kilogram of ripe fruit of high-flavonoid apples. In the step (4) of the highly alcohol-based preparation method, the addition amount of the Saccharomyces cerevisiae is: 10 ¹⁰ cfu Saccharomyces cerevisiae is added correspondingly per kilogram of fresh weight of ripe fruit of high flavonoid apples. During the fermentation process, the temperature of the fermentation system was continuously monitored 24 hours after adding Saccharomyces cerevisiae, and the temperature was controlled at 15-20°C. When said "until the alcohol content of the fermentation system no longer increases", the alcohol content of the fermentation system is 9%-12%. The Saccharomyces cerevisiae can specifically be high flavonoid cider yeast No. 1.

In the step (5) of the highly alcohol-based preparation method, the "until obtaining a distillate with an alcohol content of 80% or more" can specifically be a distillate with an alcohol content of 80%-85% . In the step (5) of the highly alcohol-based preparation method, the "until obtaining a distillate with an alcohol content of 80% or more" may specifically be to obtain a distillate with an alcohol content of 80%.

The high-flavonoid apple is an apple germplasm whose flavonoid content is more than 5000 mg per kilogram fresh weight of mature fruit. The high-flavonoid apple is an apple germplasm whose flavonoid content is more than 7000 mg per kilogram fresh weight of mature fruit. The high-flavonoid apple is an apple germplasm whose flavonoid content is more than 10000 mg per kilogram fresh weight of mature fruit. The high-flavonoid apple can specifically be 'CSR6R6-888', 'CSR6R6-666' or 'CSR6R6-777'.

'CSR6R6-888', also known as Apple (Malus domestica) CSR6R6-888, has been preserved in the General Microbiology Center of China Committee for Microbial Culture Collection (CGMCC for short) on June 16, 2017, the address is: Beichenxi, Chaoyang District, Beijing Road No. 1, Yard No. 3, Institute of Microbiology, Chinese Academy of Sciences), the deposit registration number is CGMCC NO.14295.

'CSR6R6-666', also known as Apple (Malus domestica) CSR6R6-666, has been preserved in the General Microbiology Center of the China Committee for Microbial Culture Collection (CGMCC for short) on February 17, 2017. The address is: Beichen West, Chaoyang District, Beijing Road No. 1, Yard No. 3, Institute of Microbiology, Chinese Academy of Sciences), the deposit registration number is CGMCC NO.13783.

'CSR6R6-777', also known as Apple (Malus domestica) CSR6R6-777, has been preserved in the General Microorganism Center of China Committee for Microbial Culture Collection (CGMCC for short) on December 08, 2016. The address is: Beichen West, Chaoyang District, Beijing Road No. 1, Yard No. 3, Institute of Microbiology, Chinese Academy of Sciences), the deposit registration number is CGMCC NO.12468.

High flavonoid cider yeast No. 1, the full name is Saccharomyces sp. High flavonoid cider yeast No. 1, which has been preserved in the General Microorganism Center of China Committee for Culture Collection of Microorganisms (CGMCC for short) on June 16, 2017 , address: No. 3, Courtyard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences), and the deposit registration number is CGMCC NO.14250.

Effective use of high-flavonoid apple germplasm and high-flavonoid cider processing equipment to develop apple-specific functional components, strong fruit aroma, strong irritation and liquor taste, which can meet the needs of different consumer groups. A series of high-flavonoid cider is of great significance for extending the apple industry chain, enriching the diversity of the wine market, improving the quality and efficiency of enterprise products, and meeting market and consumer demand. The dry soft high-flavonoid cider developed by the present invention not only contains flavonoids, phenolic acids and mineral elements and other apple-specific functional health-care ingredients, but also has strong irritation and taste of liquor, pure wine aroma, strong fruit aroma, The entrance is clear and refreshing, with a lingering fragrance after drinking. It is suitable for people who drink low-alcohol liquor. It has great application and promotion value and huge market prospects.

Description of drawings

Figure 1 is a photo of 'CSR6R6-888' dry soft high flavonoid cider.

Figure 2 is a photo of 'CSR6R6-888' dry soft high flavonoid cider.

Detailed ways

The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent stores. Quantitative experiments in the following examples were all set up to repeat the experiments three times, and the results were averaged.

References mentioning 'Red Fuji' and 'Gala' apples: Chen Xuesen, Xin Peigang, etc., The Role of Marshal and Jinshuai in Breeding New Apple Varieties, Journal of Shandong Agricultural University, 1994, 25(2): 236— 248.

The preparation method of 0.5% hydrochloric acid methanol solution: mix 0.5 volume part of 35% concentrated hydrochloric acid with 99.5 volume parts methanol.

The pectinases used in the examples are all pectinases purchased from Ningxia Hersbit Biotechnology Co., Ltd. The relevant parameters are: solid powder, enzyme activity ≥ 500,000 u/g, and the recommended use condition is "pH3.2- 5.0, 10-50°C". Under the conditions of 50.0°C and pH 3.5, the amount of enzyme that catalyzes the hydrolysis of pectin to generate 1 μg of galacturonic acid in 1 minute is one pectinase activity unit (1u).

Example 1. Acquisition and Identification of Excellent Apple Germplasm 'CSR6R6-888'

The method for identifying whether an apple plant is R1R1 genotype, R6R6 genotype or R6R1 genotype is as follows: take the apple from the apple plant to be tested, extract the genomic DNA of the apple pulp, use the genomic DNA as a template, and use a primer pair composed of F3 and R3 PCR amplification, then judge the genotype according to the following standards: if the PCR amplification product is a band and is 497bp, the apple plant to be tested is R6R6 genotype; if the PCR amplification product is a band and is 386bp, the apple plant to be tested is R1R1 genotype; if the PCR amplification products are two bands and are 497bp and 386bp respectively, the apple plant to be tested is R6R1 genotype.

F3 (SEQ ID NO: 1): 5'-GGTGGTCAAAGATGTGTGTTGT-3';

R3 (SEQ ID NO: 2): 5'-TTTGCCTGCTACCCACTTCA-3'.

After testing, both 'Red Fuji' and 'Gala' apples were of R1R1 genotype.

1. Acquisition of ‘CSR6R6-888’

Xinjiang red-fleshed apples were used as parents, and were crossed with white-flesh cultivated apple varieties such as 'Red Fuji'. According to Mendel's law of inheritance, when Xinjiang red-flesh apple (R6R1 genotype) crosses with white-flesh cultivars such as 'Red Fuji' (R1R1 genotype), the offspring population should be red-flesh phenotype (R6R1 genotype): white-flesh phenotype (R1R1 genotype)=1:1. However, a single plant of the R6R6 genotype was found in the hybrid _F1 population.

A single strain of the R6R6 genotype was named 'CSR6R6-888'.

'CSR6R6-888' has the following phenotype: stems, leaves, flowers, peels, and pulps, as well as various developmental stages, are dark purple.

Fresh fruit quality of ‘CSR6R6-888’: sweet and sour, crisp and juicy, good fresh quality.

The 'CSR6R6-888' was multiplied by grafting shoots or tissue culture.

2. Deposit of 'CSR6R6-888'

'CSR6R6-888', also known as Apple (Malus domestica) CSR6R6-888, has been preserved in the General Microbiology Center of China Committee for Microbial Culture Collection (CGMCC for short) on June 16, 2017, the address is: Beichenxi, Chaoyang District, Beijing Road No. 1, Yard No. 3, Institute of Microbiology, Chinese Academy of Sciences), the deposit registration number is CGMCC NO.14295.

3. Deposit of ‘CSR6R6-666’

'CSR6R6-666' is another individual plant of R6R6 genotype bred by the inventor's laboratory in the early stage.

'CSR6R6-666' has the following phenotype: stems, leaves, flowers, peels, and pulps, as well as various developmental stages, are purple-red.

'CSR6R6-666', also known as Apple (Malus domestica) CSR6R6-666, has been preserved in the General Microbiology Center of the China Committee for Microbial Culture Collection (CGMCC for short) on February 17, 2017. The address is: Beichen West, Chaoyang District, Beijing Road No. 1, Yard No. 3, Institute of Microbiology, Chinese Academy of Sciences), the deposit registration number is CGMCC NO.13783.

4. Deposit of 'CSR6R6-777'

'CSR6R6-777' is another individual plant of R6R6 genotype bred by the inventor's laboratory in the early stage.

'CSR6R6-777', also known as Apple (Malus domestica) CSR6R6-777, has been preserved in the General Microorganism Center of China Committee for Microbial Culture Collection (CGMCC for short) on December 08, 2016. The address is: Beichen West, Chaoyang District, Beijing Road No. 1, Yard No. 3, Institute of Microbiology, Chinese Academy of Sciences), the deposit registration number is CGMCC NO.12468.

5. Content Analysis of Flavonoid Components

Respectively, 'CSR6R6-888', 'CSR6R6-666' and 'CSR6R6-777' were used as the plants to be tested.

1. Take the ripe apples on the plant to be tested, and take the apple pulp.

2. Take the pulp obtained in step 1 and grind it in liquid nitrogen to obtain powder.

3. Weigh 2g of the powder obtained in step 2, add 5mL of 0.5% methanolic hydrochloric acid solution, stand at 4°C for 2h, then centrifuge at 8000rpm for 20min, and collect the supernatant and residue respectively.

4. Take the residue obtained in step 3, add 5 mL of 0.5% methanolic hydrochloric acid solution, stand at 4°C for extraction for 1 h, then centrifuge at 8000 rpm for 20 min, and collect the supernatant.

5. Mix the supernatant obtained in step 3 and the supernatant obtained in step 4 to obtain a mixed solution.

6. Take the mixed solution obtained in step 5, and remove methanol by rotary evaporation at 37°C, dissolve the residue with 2-3ml methanol, then centrifuge at 8000rpm for 20min, and collect the supernatant.

7. Take the supernatant obtained in step 6, dilute it to 5 ml with methanol, then filter it with a 0.45 μm membrane filter, and collect the filtrate.

8. The filtrate obtained in step 7 was analyzed by HPLC-MS.

Liquid chromatography conditions:

The WATERS ACQUITY UPLC chromatograph is used, the chromatographic column is BEH C18 column (100mm×2.1mm), the filler particle size is 1.7μm; the column temperature is 45°C; the injection volume is 1μL;

The mobile phase is a mixture of A liquid and B liquid, and the flow rate is 0.3mL/min; A liquid is acetonitrile, and B liquid is an aqueous solution containing 0.2% (volume fraction) formic acid; 0-0.1min, A liquid accounts for the volume of the mobile phase The fraction is 5%; 0.1-20min, the volume fraction of liquid A in the mobile phase rises linearly from 5% to 20%; 20-22min, the volume fraction of liquid A in the mobile phase rises linearly from 20% to 80%; 22-22.1 min, the volume fraction of liquid A in the mobile phase decreased linearly from 80% to 5%; 22.1-25 min, the volume fraction of liquid A in the mobile phase was 5%.

Mass Spectrometry Conditions:

The mass spectrometer is WATERS MALDI SYNAPT Q-TOF MS, ESI ionization source, electrospray ionization positive ion acquisition mode (ESI+); scanning range 100-1500m/z; capillary voltage 3.5kV, cone voltage 30V; source temperature 100°C, The desolvation temperature is 300°C; the desolvation gas flow rate is 500L/h.

The contents of 9 specific flavonol substances are shown in Table 1. The detection methods of the 9 unique substances in Table 1 belong to the detection methods of flavonoid components and content. Evaluation [J]. Chinese Agricultural Sciences, 2014, 47(11): 2193-2204.

Table 1

The above results show that 'CSR6R6-888', 'CSR6R6-666' and 'CSR6R6-777' all have strong flavonoid synthesis ability, the pulp is rich in flavonoids, and contains special flavonol components, which are high class of flavonoids. Excellent germplasm of flavonoid apple varieties. 'CSR6R6-888' outperformed 'CSR6R6-666', and 'CSR6R6-666' outperformed 'CSR6R6-777'.

6. Determination of flavonoid content

Respectively, 'CSR6R6-888', 'CSR6R6-666' and 'CSR6R6-777' were used as the plants to be tested.

(1) Take the ripe apple fruit on the plant to be tested, and take the apple pulp.

(2) Take 1g of pulp, grind it with liquid nitrogen, then add 10ml of 4°C pre-cooled 65% (volume percentage) ethanol aqueous solution and mix well, stand at 4°C in the dark for 4h, then centrifuge at 12000g for 20min, and collect the supernatant liquid.

(3) Take the test tube, add 0.5ml of the supernatant obtained in step (2), then add 1mL 5g/100ml NaNO ₂ aqueous solution, 1ml 10g/100ml AL(NO ₃ ) ₃ aqueous solution, 4mL 2M NaOH aqueous solution, and mix well Let stand for 15 minutes, centrifuge at 8000 rpm for 10 minutes, take the supernatant, and measure the absorbance at 510 nm.

A standard curve was made with rutin (Sigma chemical, ST, Loiuis, USA) as the standard sample.

The flavonoid content of 'CSR6R6-888' apple fruit was 11358.7 mg/kg fresh weight.

The flavonoid content of 'CSR6R6-666' apple fruit was 9134.6 mg/kg fresh weight.

The flavonoid content of the apple fruit of 'CSR6R6-777' was 7555.1 mg/kg fresh weight.

Embodiment 2, the acquisition of yeast strain

1. Domestication and isolation of natural yeast

When the apples growing on the 'CSR6R6-888' plants are ripe, select good apples in the orchard and bring them back to the aseptic room, cut the peel into small pieces and put them into sterilized test tubes containing juice, and plug them with cotton plugs Place them in a constant temperature incubator at 25-28°C for 5-8 days. Yeast strains were isolated by streaking on plates, and three plates were made for each fermented mash, and cultured in an incubator at 28°C for 3 days. Observe the morphology of the colonies, pick well-separated and typical single colonies from the plate, and inoculate them on the slant of the test tube and the sterilized large test tube of apple juice, and select only 3 A single colony was numbered and marked to indicate the source of the strain. Place the slant of the test tube at 28°C and incubate for 3 days to check whether the bacterial lawn is pure, and the good strains with simple bacterial lawn should be stored in the refrigerator.

2. Yeast screening

1. Duchenne tube fermentation screening.

Using the Duchenne tube fermentation method, under the same culture conditions, measure the speed of the yeast strains to produce bubbles and the number of bubbles produced within a specified time, initially compare the fermentation ability and fermentation ability of each strain of yeast, and screen out the ones with excellent fermentation performance yeast strain. The experiment was repeated 3 times in parallel. Conditions for strain activation: 24 hours of constant temperature cultivation in apple juice at 10°Brix at 25°C. Fermentation conditions: Static fermentation in apple juice at 15°Brix at 20°C for 48h.

2. Yeast fermentation test (CO ₂ weight loss method).

3. Comparison of yeast cohesion (comparison method of yeast number).

4. Ethanol resistance and SO ₂ resistance test of yeast.

Using the Dunbar tube fermentation method, the yeast were respectively inserted into different ethanol concentrations (6%, 8%, 10%, 12%, V/V) and different _SO2 concentrations (50mg/L, 100mg/L, 150mg/L, 200mg/L) of apple juice, cultivated under the same conditions, observed the generation of bubbles in the Duchenne tube, compared the tolerance of each yeast strain to ethanol and SO ₂ , and further determined the strains suitable for cider brewing. The experiment was repeated 3 times in parallel. Conditions for strain activation: 24 hours of constant temperature cultivation in apple juice at 10°Brix at 25°C. Fermentation conditions: Static fermentation in apple juice at 15°Brix at 20°C for 96h (due to the presence of ethanol or SO ₂ , the fermentation of yeast will be inhibited to varying degrees, so the gas production observation time is extended to 96h). Inoculum size: (6~8)×10 ⁷ cells/ml.

5. Fermentation test of cider brewed by yeast.

Take a 500ml Erlenmeyer flask, add 400ml of apple juice (sugar content 20°Brix), inoculate 20ml/L of yeast seed liquid with a bacterial concentration of (3-3.6)× ¹⁰⁶ /ml, and ferment at 20°C. The cider was fermented to the 15th day and the tank was inverted to remove the wine legs. After aging for 10 days, the physical and chemical indicators of the cider were measured and sensory analysis was carried out.

Through the comparison of yeast fermentation ability, cohesion, resistance to SO ₂ and ethanol, and the analysis of physical and chemical indicators and sensory quality evaluation of the fermented cider, three strains of the best cider Saccharomyces cerevisiae were selected and named as high flavonoid Apple Cider Yeast No. 1, High-Flavonoid Apple Cider Yeast No. 2, High-Flavonoid Apple Cider Yeast No. 3.

Three, compare the performance of the three yeast strains that step 2 obtains to produce cider

The yeasts to be tested are: high flavonoid cider yeast No. 1, high flavonoid cider yeast No. 2 or high flavonoid cider yeast No. 3.

1. Take 'CSR6R6-888' ripe fruit without rot and wash it thoroughly with tap water.

2. After completing step 1, take the fruit, crush and beat until the pulp particle size is below 1mm.

3. After step 2 is completed, the pulp is sterilized (80°C, 12 hours).

4. After completing step 3, cool naturally, and add pectinase when the pulp temperature is 50° C. (add 2-3 g of pectinase per 1000 g of fresh weight fruit).

5. After completing step 4, continue to cool naturally. When the temperature of the fruit pulp is 30° C., add the yeast to be tested (10 ¹⁰ cfu of the yeast to be tested is added for every 1000 g of fresh weight fruit) to carry out fermentation. During the fermentation process, the alcohol content of the fermentation system is continuously monitored, and the fermentation is stopped when the alcohol content no longer increases (the alcohol content of the system at this moment is 9%-12%). During the fermentation process, the temperature of the fermentation system was continuously monitored 24 hours after adding the yeast to be tested, and the temperature was controlled at 15-20°C.

Analyze the physical and chemical indicators and evaluate the sensory quality of the obtained cider. The indicators of high-flavonoid cider yeast No. 1 are the most ideal. It can withstand 12% ethanol. The alcohol content of the original wine fermented by it reaches 11.9%, and the wine body is clear and transparent , with the typical flavor of cider.

4. Identification of high flavonoid cider yeast No.1

Morphological characteristics: ellipsoid, with obvious nuclei and vacuoles of various sizes.

Physiological and biochemical characteristics: can use glucose, maltose, fructose, soluble starch, sucrose, galactose to ferment gas, can assimilate glucose, maltose, fructose, soluble starch, sucrose, galactose, rhamnose, cellobiose, citric acid, Can not produce acid and starch-like compounds, can produce ester aroma substances.

Growth characteristics: the optimum growth pH is 5.5-6.5, and the optimum growth temperature is 29-31°C.

5. Preservation of high flavonoid cider yeast No. 1

High flavonoid cider yeast No. 1, the full name is Saccharomyces sp. High flavonoid cider yeast No. 1, which has been preserved in the General Microorganism Center of China Committee for Culture Collection of Microorganisms (CGMCC for short) on June 16, 2017 , address: No. 3, Courtyard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences), and the deposit registration number is CGMCC NO.14250.

Example 3 and Example 4 were carried out using high flavonoid cider yeast No. 1 as Saccharomyces cerevisiae.

Embodiment 3, the preparation of high alcohol base

Apples from ‘CSR6R6-888’, apples from ‘CSR6R6-666’ and apples from ‘CSR6R6-777’ were used to prepare high alcohol bases.

1. Take the ripe apple fruit without rot, and wash it thoroughly with tap water to remove impurities.

2. After completing step 1, take the fruit, crush and beat until the pulp particle size is below 1 mm.

3. After completing step 2, sterilize the pulp (100°C, 5 hours; in practical applications, sterilize at 80-100°C for 5-12 hours, and use a longer sterilization time when the temperature is low. When the temperature is higher, a shorter sterilization time is used, for example, when the temperature is 80°C, the sterilization time is 10-12 hours, and when the temperature is 100°C, the sterilization time is 5 hours).

4. After completing step 3, cool naturally. When the temperature of the pulp is 45°C (in practical applications, 45-55°C can be used, specifically 45-50°C), add pectinase (per 1000g fresh weight fruit , add 2-3g pectinase; the enzyme activity of 2-3g pectinase is more than 1 million u-1.5 million u).

5. After completing step 4, continue to cool naturally. When the pulp temperature is 30°C (in practical applications, 25-35°C can be used), add Saccharomyces cerevisiae (add 10 ¹⁰ cfu Saccharomyces cerevisiae per 1000g fresh weight fruit) , to ferment. During the fermentation process, the alcohol content of the fermentation system is continuously monitored, and the fermentation is stopped when the alcohol content no longer increases (the alcohol content of the system at this moment is 9%-12%). During the fermentation process, the temperature of the fermentation system was continuously monitored 24 hours after adding Saccharomyces cerevisiae, and the temperature was controlled at 15-20°C.

6. After completing step 5, carry out distillation and collect the distillate between 85-95°C. The alcohol content of this distillate is about 20%.

7. Take the distillate obtained in step 6, carry out multiple distillations, and collect the distillate between 85-95°C each time until the distillate with an alcohol content of more than 80% is obtained, which is a high alcohol base .

The high alcohol base obtained from 'CSR6R6-888' is named as 'CSR6R6-888' high alcohol base. The alcoholic strength of ' CSR6R6-888 ' high alcohol base that present embodiment obtains is 80%.

The high alcohol base obtained from 'CSR6R6-666' is named as 'CSR6R6-666' high alcohol base. The alcoholic strength of ' CSR6R6-666 ' high alcohol base that present embodiment obtains is 80%.

The high alcohol base obtained from 'CSR6R6-777' is named as 'CSR6R6-777' high alcohol base. The alcoholic strength of ' CSR6R6-777 ' high wine base that present embodiment obtains is 80%.

Embodiment 4, the preparation of low alcohol base

Apples from ‘CSR6R6-888’, apples from ‘CSR6R6-666’ and apples from ‘CSR6R6-777’ were used to prepare low-alcohol wine bases.

1. Take the ripe apple fruit without rot, and wash it thoroughly with tap water to remove impurities.

2. After completing step 1, take the fruit, crush and beat until the pulp particle size is below 1mm.

3. After completing step 2, sterilize the pulp (100°C, 5 hours; in practical applications, sterilize at 80-100°C for 5-12 hours, and use a longer sterilization time when the temperature is low. When the temperature is higher, a shorter sterilization time is used, for example, when the temperature is 80°C, the sterilization time is 10-12 hours, and when the temperature is 100°C, the sterilization time is 5 hours).

4. After completing step 3, cool naturally. When the temperature of the pulp is 45°C (in practical applications, 45-55°C can be used, specifically 45-50°C), add pectinase (per 1000g fresh weight fruit , add 2-3g pectinase; the enzyme activity of 2-3g pectinase is more than 1 million u-1.5 million u).

5. After completing step 4, continue to cool naturally. When the pulp temperature is 30°C (in practical applications, 25-35°C can be used), add Saccharomyces cerevisiae (add 10 ¹⁰ cfu Saccharomyces cerevisiae per 1000g fresh weight fruit) , to ferment. During the fermentation process, the alcohol content of the fermentation system is continuously monitored, and the fermentation is stopped when the alcohol content no longer increases (the alcohol content of the system at this moment is 9%-12%). During the fermentation process, the temperature of the fermentation system was continuously monitored 24 hours after adding Saccharomyces cerevisiae, and the temperature was controlled at 15-20°C.

6. After completing step 5, pass through a 600-mesh sieve to collect the liquid phase, which is the low-alcohol alcohol base.

The low-alcohol base obtained from 'CSR6R6-888' is named as 'CSR6R6-888' low-alcohol base. The alcoholic strength of ' CSR6R6-888 ' low alcohol base that present embodiment obtains is 10%.

The low-alcohol base obtained from 'CSR6R6-666' is named as 'CSR6R6-666' low-alcohol base. The alcoholic strength of ' CSR6R6-666 ' low-alcohol wine base that present embodiment obtains is 10%.

The low-alcohol base obtained from 'CSR6R6-777' is named as 'CSR6R6-777' low-alcohol base. The alcoholic strength of ' CSR6R6-777 ' low alcohol base that present embodiment obtains is 10%.

Embodiment 5, the preparation of dry soft type high flavonoid cider

1. Mix 1 part by volume of high-alcohol base and 2 parts by volume of low-alcohol base, first let stand at 30°C for 24 hours, then stand at 0°C for 48 hours, then pass through a 600-mesh sieve to collect the liquid phase, which is the first liquor. In practical applications, it can be left to stand at 25 to 45°C for 24 to 72 hours, and then at 0 to 4°C for 24 to 72 hours.

2. Take the initial wine obtained in step 1, and carry out high and low temperature alternating aging (first stand at 30°C for 24 hours, then at -10°C for 48 hours, repeat 15 times), to obtain finished wine. In practical applications, it can be left to stand at 25 to 45°C for 24 to 72 hours first, then at -13°C to -7°C for 24 to 72 hours, and repeated more than 15 times.

The finished wine prepared by using the 'CSR6R6-888' high-alcohol base prepared in Example 3 and the 'CSR6R6-888' low-alcohol base prepared in Example 4 is named 'CSR6R6-888' dry soft high flavonoid cider. Photos of ‘CSR6R6-888’ dry soft high-flavonoid cider are shown in Figure 1 and Figure 2.

The finished wine prepared by using the 'CSR6R6-666' high alcohol base prepared in Example 3 and the 'CSR6R6-666' low alcohol base prepared in Example 4 is named 'CSR6R6-666' dry soft high flavonoid cider.

The finished wine prepared by using the 'CSR6R6-777' high alcohol base prepared in Example 3 and the 'CSR6R6-777' low alcohol base prepared in Example 4 is named 'CSR6R6-777' dry soft high flavonoid cider.

Embodiment 6, alcohol content detection and sensory description

Get three kinds of dry and soft high-flavonoid cider prepared in Example 5, and detect the alcohol content and sensory index.

The results are shown in Table 2. The dry soft high-flavonoid cider with an alcohol content of 33% is orange-red in appearance, clear and transparent, with a pure and fruity aroma, mellow, refreshing and refreshing after drinking.

Table 2

Embodiment 7, detection of relevant indicators of national food safety standards

Three kinds of dry soft high-flavonoid cider prepared in Example 5 were taken, and the contents of methanol and heavy metals were detected according to the national food safety standards GB2757-2012 and GB2760-2014.

The results show that the content of methanol in the three dry and soft high-flavonoid ciders prepared in Example 5 meets the national standards for food safety indicators, and the content of aluminum and manganese in the three dry and soft high-flavonoid ciders prepared in Example 5 The contents of heavy metals are in line with the national standards of food safety indicators.

Embodiment 8, detection of nutrition and health care ingredients

The 38° Taishan Tequ Baijiu produced by Shandong Tai'an Distillery was used as a control. The flavonoid content, phenolic acid content and mineral element content of the three dry soft high-flavonoid ciders prepared in Example 5 and the control wine were detected respectively.

1. Extraction

1. Take 20mL of wine to be tested, adjust the pH value to 7 with 1mol/L NaOH, and extract with ethyl acetate for 3 times (add 20mL of ethyl acetate each time, mix well, let it stand and separate, and then collect the organic phase). The organic phases collected by three extractions were mixed, and then the solvent was rotatably evaporated at 40°C. The residue obtained at this time was flavonoids (neutral phenols), and 2 mL of methanol was added to dissolve the residue, then filtered with a 0.22 μm filter membrane, and then determined by HPLC .

2. Take the remaining wine liquid after 3 extractions in step 1, adjust the pH value to 2 with 2mol/L HCl, and extract 3 times with ethyl acetate (add 20mL ethyl acetate each time, mix well and let it stand for stratification. Then collect the organic phase), mix the organic phases collected by the three extractions, and then evaporate the solvent at 40°C, the residue obtained at this time is phenolic acid (acid phenol), add 2mL methanol to dissolve the residue, and then use a 0.22μm filter membrane Filtration followed by HPLC assay.

2. HPLC detection

1. Relevant parameters of flavonoids detected by HPLC

The high-performance liquid chromatography device model is 2695 (Waters, Milford, MA, USA), diode array detector (PDA 2998Waters, Milford, MA, USA), quarter pump and automatic sampler; the separation column is a symmetrical C 18 (4.6 × 150 mm, 3.5 μm) column (Waters, Milford, MA, USA), column temperature 20°C.

The injection volume was 10 μL.

The mobile phase is composed of solvent A (aqueous solution containing 2.5% acetic acid by volume) and solvent B (acetonitrile), or, the mobile phase is solvent B. Elution process (flow rate 1mL/min): 0-5min, the volume fraction of solvent B in the mobile phase increases linearly from 3% to 9%; 6-15min, the volume fraction of solvent B in the mobile phase increases linearly from 9% to 16% %; 16-33min, solvent B accounts for the volume fraction of mobile phase and rises to 36.4% linearly by 16%; 34-38min solvent B accounts for the volume fraction of mobile phase and keeps 100%; Last column repairs 10 minutes (solvent B accounts for mobile phase Volume fraction is 3%).

The detection wavelength of flavanol and dihydrochalcone is 280nm, and the detection wavelength of flavonol is 360nm. Flavanols include catechin, epicatechin, and proanthocyanidin B2. Dihydrochalcones include phlorizin. Flavonols include kaempferol, rutin, isoquercitrin (quercetin-3-glucoside), guacagin (quercetin-3-O-α-L-arabinopyranoside), hyperin (Quercetin-3-O-β-D-galactopyranoside), quercetin rhamnoside.

The elution time of catechin standard was 12.52min. The peak time of epicatechin standard was 16.06min. The peak eluting time of proanthocyanidin B2 standard product was 13.83min. The elution time of the standard phlorizin was 27.76min. The peak time of kaempferol standard product was 36.86min. The elution time of standard rutin was 21.37min. The peak eluting time of isoquercitrin standard substance was 23.25min. The peak eluting time of the standard guavaside was 24.39min. The peak eluting time of hyperin standard substance was 22.64min. The peak eluting time of quercetin rhamnoside standard substance was 25.35min. All the above standard products were purchased from Sigma-Aldrich Co.

2. Relevant parameters for HPLC detection of phenolic acids

The high-performance liquid chromatography device model is 2695 (Waters, Milford, MA, USA), diode array detector (PDA 2998Waters, Milford, MA, USA), quarter pump and automatic sampler; the separation column is a symmetrical C 18 (4.6 × 150 mm, 3.5 μm) column (Waters, Milford, MA, USA), column temperature 30°C.

The injection volume was 10 μL.

The mobile phase consisted of acetonitrile and 0.6% (volume ratio) acetic acid in water. Elution process (flow rate 0.5mL/min): 0-35min, the volume fraction of acetonitrile in the mobile phase increases linearly from 5% to 35%; 36-40min, the volume fraction of acetonitrile in the mobile phase maintains 35%; 41-42min, The volume fraction of acetonitrile in the mobile phase decreased linearly from 35% to 5%.

The detection wavelength is 280nm.

The peak eluting time of gallic acid standard was 6.51min. The peak eluting time of p-hydroxybenzoic acid standard was 15.33min. The elution time of catechin standard product was 15.55min. The elution time of chlorogenic acid standard was 15.60min. The peak time of caffeic acid standard was 17.29min. The elution time of the syringic acid standard was 17.3 minutes. The peak eluting time of vanillin standard was 21.66min. The elution time of the coumaric acid standard was 22.35 minutes. The peak eluting time of the standard ferulic acid was 24.09min. The elution time of the standard benzoic acid was 28.35min. The elution time of standard coumarin was 30.69min. The elution time of cinnamic acid standard was 37.21min. The elution time of the standard phloretin was 38.49min. All the above standard products were purchased from Sigma-Aldrich Co.

3. Mineral element detection

Instrument: Inductively coupled plasma mass spectrometry (ICP-MS, Thermo Fisher Scientific, USA); WX-8000 microwave digestion instrument (Shanghai Yiyao Instrument Technology Development Co., Ltd.); Dura series ultrapure water treatment system; the working parameters of the instrument are shown in the table 3.

table 3

project parameter project parameter RF power (W) 1050 Analog voltage (V) -1900 Atomizing gas flow rate (L/min) 0.86 Pulse voltage (V) 900 Sample lifting rate (L/min) 1.2 Data collection mode peak jump Ion lens voltage (V) 8.25 Dwell time (ms) 50 Cooling air flow(L/min) 13.8 Collect data points per mass 3 Carrier gas flow(L/min) 0.98 Integration time (ms) 500

Each single element standard solution (National Nonferrous Metals and Electronic Materials Analysis and Testing Center, 1000μg/mL). Preparation of standard solution (using 5% _HNO3 aqueous solution as solvent): absorb single-element standard solution, dilute to 20mg/L stock solution, then dilute to 200μg/L, 400μg/L stock solution, and then prepare in turn to obtain 8μg/L A series of standard solutions of L, 16μg/L, 24μg/L, 32μg/L, 48μg/L, 64μg/L, shake well and set aside. 5% HNO ₃ aqueous solution was used as the 0-concentration marking solution. The standard solution was detected by mass spectrometry to make a standard curve.

Weigh 0.2-1g of the wine sample to be tested into a polytetrafluoroethylene digestion tank (accurate to 0.1mg), add 5mL of concentrated nitric acid (the mass fraction of concentrated nitric acid is about 68%), and then put the digestion tank into a microwave digestion instrument, The digestion procedure is shown in Table 4. Then cool naturally. After the temperature has cooled to below 50°C, take out the digestion tank and put it in a fume hood, open the digestion tank, rinse with ultrapure water, and then transfer the liquid phase in the digestion tank to a 50mL volumetric flask. Dilute with pure water to volume, and then perform mass spectrometry detection. The content of each single element in the wine sample to be tested was calculated according to the standard curve.

Table 4

step temperature(℃) Holding time (min) 1 100 3 2 140 3 3 160 3 4 180 3 5 190 15

4. Results

The contents of flavonoid components in three dry soft high-flavonoid ciders and the control wine are shown in Table 5. The content of phenolic acid components in three dry soft high-flavonoid ciders and the control wine is shown in Table 6. The contents of mineral elements in three dry soft high-flavonoid ciders and the control wine are shown in Table 7. Contents of flavonoid components, phenolic acid components and The content of mineral elements was significantly higher than that of the control wine with similar degrees, among which the content of flavonoid components, phenolic acid components and mineral elements in dry soft high-flavonoid cider made from ripe apple fruits of CSR6R6-888 Highest.

Table 5 flavonoid component content

Table 6 phenolic acid component content

Table 7 Mineral element content

SEQUENCE LISTING

<110> Shandong Agricultural University

<120> dry soft high-flavonoid cider and preparation method thereof

<130> GNCYX171210

<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 accacttca 20

Claims (6)

1. A preparation method for cider, comprising the steps of: ①Mix 1 volume part of high-alcoholic alcohol base with 1.8-2.2 volume parts of low-alcohol alcohol base, first stand at 25-45°C for 24-72 hours, then at 0-4°C for 24-72 hours, and collect the liquid phase, which is First wine; ② Take the initial wine obtained in step ①, and carry out high and low temperature alternate aging to obtain the finished wine; The low-alcohol alcohol base uses the ripe fruit of high-flavonoid apples as the only raw material, and the preparation method includes the following steps in sequence: (1) crushing and beating; (2) Sterilization; (3) Add pectinase; (4) Add Saccharomyces cerevisiae and carry out fermentation until the alcohol content of the fermentation system no longer increases; (5) Collect the liquid phase, which is the low-alcohol alcohol base; The high-alcoholic alcohol base uses the ripe fruit of high-flavonoid apples as the only raw material, and the preparation method includes the following steps in sequence: (1) crushing and beating; (2) Sterilization; (3) Add pectinase; (4) Add Saccharomyces cerevisiae and carry out fermentation until the alcohol content of the fermentation system no longer increases; (5) Distill and collect the distillate between 85-95°C, then distill the distillate again and collect the distillate between 85-95°C, several times until the distillate with an alcohol content of 80% or more is obtained The product is a high alcohol base; In the low-alcohol wine base and the high-alcohol wine base, the high-flavonoid apple as the only raw material is 'CSR6R6-888'; the full name of 'CSR6R6-888' is Apple ( Malus domestica ) CSR6R6-888, and its preservation registration The number is CGMCCNO.14295; In the preparation method of the low alcohol base and the high alcohol base, the Saccharomyces cerevisiae is high flavonoid cider yeast No. 1; the full name of high flavonoid cider yeast No. 1 is Saccharomyces sp. High-flavonoid cider yeast No. 1, its deposit registration number is CGMCC NO.14250. 2. The method according to claim 1, characterized in that: in the step (1), crushing and beating until the pulp particle size is below 1mm. 3. The method according to claim 1, characterized in that: in the step (3), when the temperature of the pulp is 45-55°C, pectinase is added. 4. The method according to claim 1, characterized in that: in the step (4), when the temperature of the pulp is 25-35°C, Saccharomyces cerevisiae is added. 5. The method according to claim 1, characterized in that: in the step ②, the alternating high and low temperature curing is as follows: first stand at 25-45°C for 24-72 hours, then stand at -13°C--7°C Leave it for 24 to 72 hours, and repeat it more than 15 times. 6. The cider prepared by the method according to any one of claims 1 to 5.