CN105901212B - Docynia delavayi fermented tea and preparation method thereof - Google Patents
Docynia delavayi fermented tea and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/34—Tea substitutes, e.g. matè; Extracts or infusions thereof
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
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- Tea And Coffee (AREA)
Abstract
The invention provides docynia delavayi
The preparation method of the fermented tea comprises the following steps: a. preparation of Docynia delavayi
A tea crude product; b. obtaining docynia delavayi of step aTreating the crude tea with steam for 10min, controlling the water content of tea pile to 12-20%, and performing pile fermentation at 45-55 deg.C for 3-5 hr; steaming for 20 min; c. fermentation of eurotium cristatum: adding sterile water according to the water-material ratio of 20-40% (mL/g), and inoculating eurotium cristatum; then culturing and fermenting for 7d-10d at the temperature of 25-35 ℃; d. c, sterilizing and drying the fermentation product in the step c to obtain docynia delavayi
Fermenting the tea. Docynia delavayi of the invention
The fermented tea has good sensory quality and taste, high essential amino acid content, high phloretin content, rich nutrient components and good application prospect.
Description
Technical Field
The invention belongs to the technical field of tea making and the technical field of microbial fermentation, and particularly relates to a specific product Docynia delavayi Durcz in substitutional tea
Fermented tea and its preparation methodA method. The product of the technology can be used for developing foods and functional foods.
Background
Docynia delavayi
Commonly known as wild apple, hawthorn tree, docynia delavayi and docynia delavayi
Docynia delavayi]Docynia delavayi 26509Docynia delavayi
Etc. of Docynia delavayi of Maloideae of Rosaceae
The general term for plants belonging to Docynia, an evergreen or semi-evergreen tree, includes three species: docynia delavayi Franch
(D.delavayi (Franch.) Schneid), Docynia delavayi
(D.indica (Wall.) Dcne) and Docynia delavayi
(D.longiunguis Q.Luo et J.L.Liu), mainly distributed in the southwest minority region and southeast Asia region of China, and originated in hillside, stream side and forest with elevation of 2000-.
Docynia delavayi
Is a common plant resource used as both medicine and food in the southwest minority nationality areas of China. The stem leaves and the fruits of the Chinese herbal medicine can be used as the medicine, and have the effects of diminishing inflammation, setting bone, relaxing muscles and joints, promoting blood circulation, soothing liver, relieving pain, clearing summer heat, disinfecting and the like; tender stem tips are also one of wild vegetables collected by Dai nationality in daily life. In addition, Docynia delavayi
Fruit nutritionRich and unique taste, is often eaten as wild fruits, and is partially developed into preserved fruits, fruit vinegar, fruit wine and the like; docynia delavayi Durcz in the current market
Most of the development and utilization are fruit sale and processing.
Studies have shown that docynia delavayi
The leaf contains high polyphenol, and has high health promotion value
The leaves can be collected all the year round, and the resource amount is large. However, Docynia delavayi is not developed and utilized at present
Leaf reports, not to mention
The preparation of leaves as substitute tea is reported.
Disclosure of Invention
The invention aims to provide docynia delavayi
Fermented tea and its preparation method are provided.
The invention provides a preparation process and a method of novel substitute leaf tea, enriches preparation technologies of substitute tea and microbial treatment technologies of raw materials, and develops development technologies of medicinal and edible dual-purpose articles.
The invention provides docynia delavayiThe preparation method of the fermented tea comprises the following steps:
a. preparation of Docynia delavayi
Tea crude product: docynia delavayi DielsSpreading fresh leaves for 2-4hr, deactivating enzyme, rolling for the first time, spreading for 0.5-1hr, rolling for the second time, and drying to water content of 8-11% to obtain Docynia delavayi
A tea crude product;
wherein the water-removing temperature is 200-300 ℃, the water-removing time is 5-10 min, the mixture is spread for 30-60 min after being taken out of the pot, and then the mixture is twisted;
wherein, the rolling is carried out by a rolling machine in a light pressing way for 10-25 min for the first time; the secondary rolling mode is light pressing for 5min-10 min;
b. obtaining docynia delavayi of step a
Treating the crude tea with steam for 10min, controlling the water content of tea pile to 12-20%, and performing pile fermentation at 45-55 deg.C for 3-5 hr; steaming for 20 min;
c. fermentation of eurotium cristatum: adding sterile water according to the water-material ratio of 20-40% (mL/g), and inoculating eurotium cristatum; then culturing and fermenting for 7d-10d at the temperature of 25-35 ℃;
d. c, sterilizing and drying the fermentation product in the step c to obtain docynia delavayi
Fermenting the tea.
Pile fermentation temperature: 45-55 ℃ means that the environment of the tea pile and the temperature of the raw materials of the tea pile are kept at 45-55 ℃.
Wherein, in the step b, the primary steam treatment is saturated steam treatment.
Saturated steam: that is, vapor in a saturated state, when liquid evaporates in a limited closed space, liquid molecules enter the upper space through the liquid surface, and when the number of molecules entering the space per unit time is equal to the number of molecules in the returned liquid, evaporation and condensation are in a dynamic equilibrium state, and the vapor at this time is called saturated vapor.
Wherein, in the step c, sterile water is added according to the water-material ratio of 35 percent.
Wherein, in the step c, after the Eurotium cristatum is inoculated, the quantity of the Eurotium cristatum is 106～107one/mL.
Wherein, in the step c, fermentation is carried out at the temperature of 28 ℃; and/or turning over for 2-3 times during the fermentation process.
Wherein in the step d, the sterilization method comprises the following steps: keeping at 60-80 deg.C for 0.5hr-1hr while turning over for 2-3 times.
Wherein, in the step d, the drying temperature is 40-60 ℃, and the water content is controlled to be 8-11%.
Further, the temperature for drying is 50 ℃.
The invention also provides the docynia delavayi prepared by the method
Fermenting the tea.
Docynia delavayi of the invention
The fermented tea has good sensory quality and taste, high essential amino acid content, high phloretin content and rich nutrient components, is suitable for being drunk widely and daily, and has higher development and application values.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 is a graph of the response signals of each sensor to a tea sample
FIG. 2 various pairs of sensorsSample response values before and after tea fermentation
Docynia delavayi Franch FIG. 3
Two-dimensional graph for principal component analysis before and after tea fermentation
Docynia delavayi (figure 4)
Three-dimensional graph for analyzing main components before and after tea fermentation
Docynia delavayi Franch FIG. 5
Common peak fingerprint before and after tea fermentation
Figure 6 measurement of Docynia delavayiChromatogram of phlorizin and phloretin content in tea
Detailed Description
The following examples are further illustrative, but the present invention is not limited to these examples.
The experimental materials and instruments used in the invention are as follows:
docynia delavayiLeaf: collected from Dehong Zhou, Panzhihua city, West Chang city, etc. of Yunnan province;
eurotium cristatum (eurotium cristatum): is purchased from Beinanjianglian institute of biotechnology, BNCC146559, namely Eurotium cristatum strain with the preservation number of China general microbiological culture Collection center of CGMCC 3.448;
the 6cr-25 model tea rolling machine is purchased from Fei Asia mechanical manufacturing company, Inc. in Jingun county of Sichuan province, α -ASTREE electronic tongue is purchased from French Alpha MOS company, Agilent 1260 high performance liquid chromatograph is purchased from Agilent, Inc. of America, S-433D amino acid analyzer is purchased from Sykam company of Germany, WSL-2 comparative colorimeter is purchased from Shanghai Xinrui instrument and balance, Inc., Sartorius BP211D model electronic chromatograph is purchased from Beijing Saedolis instrument and system, Inc., and DHG-9240A electrothermal constant temperature air-blast drying box is purchased from Shanghai Jing Macro Experimental facilities, Inc.
Example 1 Docynia delavayi of the invention
Preparation of fermented tea
Preparation of unfermented Docynia delavayi
Tea crude product
According to the pan-frying process of green tea, the Docynia delavayi is prepared
The tea is prepared by the following specific method:
docynia delavayi Diels
Spreading fresh leaves for 2-4hr, deactivating enzyme, rolling for the first time, spreading for 0.5-1hr, rolling for the second time, drying, controlling water content at 8-11%, and docynia delavayi
A tea crude product;
the deactivation of enzymes and the rolling are key links:
1) the water-removing temperature is 200-300 ℃, and the water-removing time is 5-10 min. The green-removing can be carried out manually by using a frying pan, the temperature of the frying pan is 200-300 ℃, the leaf feeding amount of each pan is 5-7 Kg, and the sound of crisp leaves like fried beans can be heard after the leaves are put into the pan. The enzyme deactivation time is 5-10 min, the enzyme deactivation time is until the green taste disappears, the faint scent overflows slightly, the leaf color is dark green, the leaf stalks are continuously folded, the enzyme deactivation leaves are pinched into a ball by hands, and the ball can be slowly bounced off after the hands are loosened. Spreading for 30-60 min after taking out of the pot, and then rolling;
2) the rolling can be carried out by a small rolling machine (for example: 6cr-25 tea rolling machine, Feiya machine manufacturing Co., Ltd., Jing, county, of Sichuan province), 15Kg-20Kg of tea leaves were put, and the tea leaves were first rolled and kneaded under light pressure for 10min-25 min. Kneading for 5-10 min under light pressure.
Preparation of Eurotium cristatum seed liquid
Inoculating eurotium cristatum strain stored at 4 deg.C on Sasa liquid culture medium, shake culturing at 28 deg.C and 150r/min for 24 hr, selecting appropriate amount of mycelium, inoculating on Sasa slant culture medium, culturing for 7d, adding appropriate amount of sterile water into slant tube, repeatedly purging slant, making spore suspension, transferring into 100mL sterile triangular flask, and regulating spore concentration to 10 by blood count plate method6～107And (5) obtaining the seed liquid after seed treatment per mL.
Thirdly, preparing docynia delavayi by solid state fermentation method
Fermented tea
The technical route is as follows: docynia delavayi
Tea crude product → steaming for 10min → pile fermentation at 45-55 deg.C for 3hr-5hr → adjusting water-material ratio to 35% → sterilizing → inoculating 5% eurotium cristatum → flowering culturing at 20-30 deg.C for 7d-10d → baking at 60-80 deg.C for 0.5hr-1hr → drying at 40-60 deg.C → product
The method comprises the following specific steps:
weighing 5g of Docynia delavayi
Steaming tea for 10min, transferring into 100mL fermentation bottle, performing pile fermentation at 45-55 deg.C for 3-5 hr (controlling water content of pile fermentation at 12-20%), performing saturated steaming for 20min, adding sterile water at water-material ratio of 20-40% (V/M), inoculating 3-10% (V/M) inoculating strain to obtain Eurotium cristatum seed liquid (controlling spore concentration of Eurotium cristatum liquid to 10)6～107one/mL), fermenting for 7d-10d at 25-35 ℃ until the surface of the tea leaves has typical yellow golden flowers, and turning over for 2-3 times in the fermentation process;
baking at 60-80 deg.C for 0.5hr-1hr while turning over for 2-3 times; finally drying at 40-60 ℃, controlling the water content to be 8-11 percent, obtaining Docynia delavayi
Fermented tea, i.e. the inventionDocynia delavayi
Tea is prepared from folium Camelliae sinensis.
The following test examples are used to illustrate the advantageous effects of the present invention:
experimental example 1 Docynia delavayi
Comparative test of tea preparation method
First, test materials
The method of the invention prepares Docynia delavayi
Tea (a): docynia delavayi prepared according to the method of example 1
Tea;
preparation of Docynia delavayi by traditional method
Tea (B): according to the pan-frying process of green tea, the Docynia delavayi is prepared
Tea (i.e., non-fermented Docynia delavayi of example 1)
Crude tea).
And (3) statistics and analysis: all experimental data were statistically analyzed using SPSS 17.0 software and Excel.
Second, measurement and evaluation test of sensory index
Will docynia delavayi
Tea and Docynia delavayi
Grinding fermented tea, sieving with 60 mesh sieve to obtain tea sample powder, packaging into tea bags according to the specification of 3 g/bag, drying and preserving.
Soaking the two samples in 1 bag of tea water at a ratio of 1: 50 respectively for 5min, and taking out the tea bag to obtain tea soup for use.
1. Determination of tea soup chroma
The chroma of the two tea soups is respectively measured by a WSL-2 comparison color measuring instrument (WSL-2 comparison color measuring instrument), and then the color of the tea soups is named according to the requirements of the instruction of the instrument.
Comparison of Docynia delavayi before and after fermentation
The color of the tea broth of the tea changed and the results are shown in Table 1.
TABLE 1 Docynia delavayi
Color change of tea soup before and after tea fermentation
Table 1Chroma changes ofnative and fermented Docyniaindica tea
As can be seen from Table 1, the color of the tea soup before and after fermentation changes obviously, the tea soup after fermentation is orange yellow, and the color comparison result of the tea soup is as follows: yellow 14, red 3.2; the color before fermentation is bright orange yellow, and the result of color comparison of the tea soup is as follows: yellow 10, red 2, grey 1.
Docynia delavayi of the invention
The color of the fermented tea soup is thicker and heavier, and is due to the fact that various active and colored substances are dissolved into the tea soup through the fermentation of the eurotium cristatum, and docynia delavayiThe color and quality of the tea are obviously improved.
Thus, Docynia delavayi obtained by the present invention
Fermentation ofThe tea has thicker and more intense color and luster quality than the non-fermented Docynia delavayiThe light color of tea is better.
2. Sensory evaluation analysis
Randomly selecting 10 sensory panel examiners, and carrying out simple food sensory training on the sensory panel examiners. And performing descriptive evaluation and grade grading on the taste and the aroma of the tea soup according to the evaluation standard in the table 2.
TABLE 2 Docynia delavayi
Criteria for tea evaluation
Table2The sensory evaluation criteria of D.indicatea
Docynia delavayi
The changes of sensory evaluation before and after tea fermentation are shown in Table 3.
TABLE 3 Docynia delavayi
Sensory evaluation results before and after tea fermentation
Table3 Results for the sensory evaluation of native and fermentedD.indica tea
Note: compared with the pre-fermentation, p is less than 0.01, and the difference is very obvious.
As can be seen from Table 3, Docynia delavayi
The sensory difference before and after the tea fermentation is very obvious (p is less than 0.001). Wherein Docynia delavayi of the present invention
The total score of the fermented tea is higher, and the sensory evaluation is obviously better than that of the non-fermented Docynia delavayiTea is prepared from folium Camelliae sinensis.
In particular, Docynia delavayi of the invention
The fermented tea has thick and heavy color and quality, the tea soup tastes sweet, and the tea soup has mellow taste which cannot be achieved before fermentation; furthermore, Docynia delavayi of the present invention
The fermented tea has unique 'fungus flower fragrance', pure and high fragrance.
3. Electronic tongue evaluation analysis
Separately separating the fermented docynia delavayi
Tea and unfermented Docynia delavayi
And filtering the two tea soups, cooling to room temperature, transferring 80mL of the two tea soups, placing the two tea soups in a 120mL electronic tongue special beaker, and standing for 15 min. The instrument was calibrated using chilled boiling water as the calibration solution, and then the tea soup was subjected to electronic tongue analysis. The electronic sensor collected 120s per tea soup sample and recorded 1 time per second. Each sample was measured in 6 replicates.
3.1 determination of sample time
According to the working principle of the electronic tongue, the electronic tongue can be subjected to self-checking, diagnosis, correction and other steps before signal collection, so that the fluctuation is large in the initial stage of detecting a sample, as shown in fig. 1.
As can be seen from FIG. 1, the horizontal axis in the graph is the measurement time, the vertical axis is the collected response value, the 7 sensors are ZZ, BA, BB, CA, GA, HA and JB respectively, and the 7 sensors are not specific sensors, have sensitivity to four tastes of sour, sweet and bitter, but have different sensitivity degreesThis corresponds to 7 panelists all evaluating the samples for overall flavor. Docynia before fermentation
For example, the tea shows that the response value changes very obviously in the first 10s, the fluctuation is obvious in the range of 10s to 50s, the fluctuation is small in the range of 50s to 100s, and the response value is basically stable in the range of 100s to 120s, so that the data of 120s is selected as the final response value. The fluctuation of the measured data of the first 3 times is large, so the measured data of the last 3 times is selected as the original data to be analyzed.
3.2 sensor response Signal analysis
And selecting the measurement data of 120s as a final response, analyzing the response signals of the sensors, and analyzing the response values of the sensors to obtain electronic tongue radar images of the samples before and after fermentation, wherein the result is shown in figure 2.
As can be seen from FIG. 2, all the sensor response values are between 150 and 8500, wherein the response values of the CA, BB, BA and HA sensors are above 4000, the response values of the rest sensors are low, but the detection signal intensity difference of the CA sensor is obvious, so that the CA is Docynia delavayi
A sensor sensitive to flavor difference of samples before and after tea fermentation.
3.3 PCA principal component analysis results
The signal collected by the electronic tongue was subjected to principal component analysis, and the results are shown in fig. 3 and 4.
Each 1 point in fig. 3 and 4 represents information for one sample, and the greater the distance between the point and the sample, the greater the difference between the samples.
From FIG. 3, Docynia delavayi
The tea soup tastes different before and after tea fermentation, the sample before fermentation is mainly concentrated on the upper part, and the sample after fermentation is concentrated on the lower part. The differences between the samples are further demonstrated from figure 4, consistent with the sensory evaluation results.
After analyzing the data, it was found that the contribution rate of principal component 1(PC1) was 95.3%, and the contribution rate of principal component 2(PC2) was 3.21%, indicating that the contribution rates of principal components 1 and 2 to the total explanatory variable were 98.5%, which sufficiently reflected the overall information of the sample.
The analysis result of the electronic tongue shows that: the tea soup before and after fermentation has obvious difference in sensory quality.
Moving three, moving
Determination of amino acid content in tea
1. Measurement method
Weighing 1.5g of samples before and after fermentation respectively, adding 80mL of boiling water, leaching in a boiling water bath for 45min, filtering, collecting the filtrate in a 100mL volumetric flask, washing residues with a small amount of deionized water, cooling, metering volume, and shaking uniformly to obtain the tea soup. Taking 5mL of tea soup, adding 5mL of 10% sulfosalicylic acid into a centrifuge tube, shaking uniformly, standing for 10min, centrifuging at 4000r/min for 10min, diluting the supernatant to different degrees, and filtering with a 0.22-micrometer microporous filter membrane to be tested. A chromatographic column: LCA K07 (150mm × 4.6mm), detection wavelength: 570nm, 440nm, eluent flow: 0.45mL/min, reaction solution flow: 0.25mL/min, column temperature: gradient temperature of 57-74 ℃, reaction temperature: 130 ℃. Comparison of Docynia delavayi
Amino acid content before and after tea fermentation, wherein:
1) content of essential amino acids for human body: the content of 8 essential amino acids in human body is the sum of the contents of threonine, methionine, valine, isoleucine, leucine, phenylalanine, lysine and tryptophan.
2) Content of flavor-developing amino acid: the content of the delicious amino acid is the sum of the contents of glutamic acid and aspartic acid; the sweet amino acid content is the sum of the glycine content, the alanine content, the serine content and the proline content; the aromatic amino acid content is the sum of tyrosine and phenylalanine.
2. Measurement results
Docynia delavayi
The results of amino acid content before and after tea fermentation are shown in Table 4.
TABLE 4 Docynia delavayiAmino acid changes before and after tea fermentation
Note: the growth rate (amino acid content after fermentation-amino acid content before fermentation)/amino acid content before fermentation × 100%.
Increased "+" amino acid content; "-" reduced amino acid content; the "0" amino acid content was unchanged.
Amino acids are important nutritional ingredients, and the content and composition of various amino acids directly affect the nutritional value of foods and are closely related to human taste.
As can be seen from Table 4, two Docynia delavayi Hemsl
Tea contains 16 major amino acids, but the amino acid content shows a large variation. Wherein Docynia delavayi of the present inventionThe essential amino acid content of the fermented tea is increased by 40.506%, and the nutritive value is higher; the content of the fresh amino acid in the flavor amino acid is reduced by 38.583 percent, while the content of the sweet amino acid and the aromatic amino acid is obviously increased by 37.838 percent and 42.775 percent respectively, thereby improving the taste as a whole.
Thus, Docynia delavayi of the present invention
The fermented tea has high content of essential amino acids, high nutritive value, and greatly improved sweet and fragrant amino acidsAnd the taste is better.
Fourthly, measuring and evaluating chemical components before and after fermentation
1. Preparation of test solution
1.1 HPLC fingerprint analysis sample preparation
Respectively removing docynia
Tea and docynia delavayi
Placing the fermented tea at 50 ℃, drying to constant weight, grinding into powder by using a mortar, sieving by using a 60-mesh sieve (the sieving rate is more than 98%), precisely weighing 0.100g of powder, placing the powder into a 25mL volumetric flask, adding 20mL of methanol, carrying out ultrasonic extraction at 25 ℃ for 30min, cooling to room temperature, fixing the volume by using the methanol to obtain an extracting solution, and passing a sample through a 0.45-micrometer microporous filter membrane to obtain a sample for fingerprint spectrum determination. Storing at low temperature for later use.
1.2 HPLC assay preparation of test samples
Precisely measuring 0.5mL of the extract obtained by filtering the extract through a 0.45-micrometer microporous membrane in the 1.1, placing the extract in a 10mL volumetric flask, adding methanol to a constant volume, and passing the sample through a 0.45-micrometer microporous membrane to obtain the sample for rapidly determining phlorizin and phloretin. Storing at low temperature for later use.
2. Sample HPLC fingerprint analysis
Comparing docynia before and after fermentation
The chemical components of tea are changed.
Chromatographic conditions are as follows:
a chromatographic column: agilent ZORBAX extended-C18 column (4.6X 250mm, 5 μm), column temperature: 25 ℃, mobile phase: acetonitrile (A), methanol (B) and water (C), and linear gradient elution: 0-40 min (5-15% A: 0-35% B: 95-50% C), 40-44 min (15-30% A: 35-0% B: 50-70% C), 44-55 min (30-40% A: 70-60% C), 55-59 min (40-100% A: 60-0% C), 59-70 min (100% A) volume flow: 1mL/min, sample size: 10 μ L, detection wavelength: 285 nm.
Comparing the Docynia delavayi according to HPLC finger prints
The chemical composition changes before and after tea fermentation, and the results are shown in FIG. 5.
As can be seen from FIG. 5, docynia delavayi before and after fermentation
The tea has a plurality of changed components, and through comparison, 18 common peaks are finally determined, and the No. 6 peak is determined to be phlorizin and the No. 10 peak is determined to be phloretin through an external standard method.
Docynia delavayi
The composition changes before and after tea fermentation are shown in Table 5.
TABLE 5 Docynia delavayi
Changes in ingredients before and after tea fermentation
Note that "+" shared peak area after fermentation increased, "-" shared peak area after fermentation decreased, "△"
A peak which disappears after fermentation, and a peak which newly appears after fermentation of '▲'.
As can be seen from table 5, of the 18 common peaks before and after fermentation, 6 chromatographic peak areas decreased and 12 chromatographic peak areas increased after fermentation; in addition, 11 chromatographic peaks appear after fermentation, which indicates that a new compound is ready, and 3 chromatographic peak areas after fermentation are reduced to zero, which indicates that a substance is completely degraded and disappears.
Thus, the Docynia delavayi of the present invention is compared to before fermentationThe chemical components of the fermented tea are obviously changed.
3. Content determination of phlorizin and phloretin in sample
Measuring the content of phlorizin and phloretin in the samples before and after fermentation, and comparing the change before and after fermentation.
3.1 drawing of Standard Curve
Precisely weighing 0.0040g of phlorizin standard substance and 0.0020g of phloretin standard substance, dissolving with methanol to constant volume to 50mL, and preparing into a reference substance mixed stock solution with phlorizin mass concentration of 80 mug/mL and phloretin mass concentration of 40 mug/mL. Transferring 0.05 mL, 0.1 mL, 0.2 mL, 0.5mL, 1, 2 mL and 5mL of stock solutions into a 10mL volumetric flask, carrying out constant volume with methanol to prepare phlorizin and phloretin mixed standard solutions with different concentrations, analyzing according to 2.3.3.1 chromatographic conditions, and carrying out linear fitting by taking the concentration X (mu g/mL) as a horizontal coordinate and the chromatographic peak area Y (mAU multiplied by min) as a vertical coordinate.
The regression equation of phlorizin is obtained as Y-19.77558X +4.22904, r20.99998, indicating that phlorizin concentrations between 0.4 and 80 μ g/mL are well linear;
the regression equation of phloretin is that Y is 34.78806X +2.96544, r20.99999, indicating a good linearity of phloretin concentration between 0.24 and 40. mu.g/mL.
3.2 measurement of content of phlorizin and phloretin in sample
Taking the prepared HPLC content determination sample, injecting sample according to the following chromatographic conditions, and calculating docynia delavayi before and after fermentation
The content of phlorizin and phloretin in the tea.
Chromatographic conditions are as follows: a chromatographic column: agilent ZORBAX extended-C18 column (4.6X 250mm, 5 μm), column temperature: 25 ℃, mobile phase: acetonitrile (A) to water (C), linear gradient elution: 0-15min (15% -50% A: 85% -50% C), 15-30min (50% -100% A: 50% -0% C), volume flow: 1mL/min, sample size: 10 μ L, detection wavelength: 285 nm.
3.3 HPLC chromatogram results of phlorizin and phloretin
Docynia delavayi
HPLC chromatogram of phlorizin and phloretin in tea are shown in figure 6.
As can be seen from FIG. 6, the Docynia delavayi of the present invention was compared with that before fermentation
The phlorizin content in the fermented tea is reduced by 27 percent (from 251.87mg/g to 198.25mg/g), while the phloretin content is increased by 6.87 times (from 2.4943mg/g to 19.619mg/g), so that the phloretin is greatly increased. Description of Docynia delavayiThe main functional components of the fermented tea are obviously changed, the phlorizin is converted into the phloretin in a large amount, and the phloretin is increased to 7.8 times of the original phloretin after being fermented for 7 days.
Phloretin is aglycon of phlorizin, has strong antioxidant activity, higher activity than phlorizin, has certain cancer adjuvant treatment effect, can be used as natural skin whitening agent, and has effects of protecting cardiovascular system, inhibiting glucose transport, and regulating autoimmunity. Docynia delavayi of the invention
The fermented tea has high content of phloretin and high nutritive and health-care value.
To sum up, docynia delavayi of the invention
The fermented tea has good sensory quality and taste, high essential amino acid content, high phloretin content, rich nutrient components and good application prospect.
Claims (9)
1. Docynia delavayiProcess for preparing fermented tea, especially fermented teaCharacterized in that: the method comprises the following steps:
a. preparation of Docynia delavayiTea crude product: docynia delavayi DielsSpreading fresh leaves for 2-4hr, deactivating enzyme, rolling for the first time, spreading for 0.5-1hr, rolling for the second time, and drying to water content of 8-11% to obtain Docynia delavayiA tea crude product;
wherein the water-removing temperature is 200-300 ℃, the water-removing time is 5-10 min, the mixture is spread for 30-60 min after being taken out of the pot, and then the mixture is twisted;
wherein, the rolling is carried out by a rolling machine in a light pressing way for 10-25 min for the first time; the secondary rolling mode is light pressing for 5min-10 min;
b. obtaining docynia delavayi of step aTreating the crude tea with steam for 10min, controlling the water content of tea pile to 12-20%, and performing pile fermentation at 45-55 deg.C for 3-5 hr; steaming for 20 min;
c. fermentation of eurotium cristatum: adding sterile water according to the water-material ratio of 20-40% (mL/g), and inoculating eurotium cristatum; then culturing and fermenting for 7d-10d at the temperature of 25-35 ℃;
d. c, sterilizing and drying the fermentation product in the step c to obtain docynia delavayiFermenting the tea.
2. The method of claim 1, wherein: in the step b, the primary steam treatment is saturated steam treatment.
3. The method of claim 1, wherein: in the step c, sterile water is added according to the water-material ratio of 35 percent.
4. The method of claim 1, wherein: in step c, after the Eurotium cristatum is inoculated, the quantity of the Eurotium cristatum is 106～107one/mL.
5. The method of claim 1, wherein: in the step c, fermentation is carried out at the temperature of 28 ℃; and/or turning over for 2-3 times during the fermentation process.
6. The method of claim 1, wherein: in the step d, the sterilization method comprises the following steps: keeping at 60-80 deg.C for 0.5hr-1hr while turning over for 2-3 times.
7. The method of claim 1, wherein: in the step d, the drying temperature is 40-60 ℃, and the moisture is controlled to be 8-11%.
8. The method of claim 7, wherein: the drying temperature is 50 ℃.
9. Docynia delavayi produced by the process of any one of claims 1 to 8Fermenting the tea.
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