CN113508839A - Compound traditional Chinese medicine lipid-lowering fermented beverage and research method of optimal proportion thereof - Google Patents
Compound traditional Chinese medicine lipid-lowering fermented beverage and research method of optimal proportion thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C2240/00—Use or particular additives or ingredients
- A23C2240/15—Use of plant extracts, including purified and isolated derivatives thereof, as ingredient in dairy products
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
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- Chemical & Material Sciences (AREA)
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Abstract
The invention discloses a compound traditional Chinese medicine lipid-lowering fermented beverage and a research method of the optimal proportion thereof, the formula components of the compound traditional Chinese medicine lipid-lowering fermented beverage comprise 30-100 parts of traditional Chinese medicine extract, 3-5 parts of glucose and 1-3 parts of skim milk powder by weight, and the research method of the optimal proportion of the compound traditional Chinese medicine lipid-lowering fermented beverage comprises the steps of determining the optimal fermentation amount of raw material proportion and determining the optimal fermentation process. In the invention, the raw material dosage and the fermentation process are optimized to obtain the optimal raw material proportion and the optimal fermentation process, which is beneficial to ensuring the taste and the production quality of the finished beverage.
Description
Technical Field
The invention relates to the technical field of fermented beverages, in particular to a compound traditional Chinese medicine lipid-lowering fermented beverage and a research method of the optimal proportion thereof.
Background
The traditional Chinese medicine decoction pieces are prepared from traditional Chinese medicines according to the traditional Chinese medicine theory and the traditional Chinese medicine preparation method, and can be directly used as traditional Chinese medicines in traditional Chinese medicine clinical practice. The traditional Chinese medicine and the traditional Chinese medicine decoction pieces have no absolute limit, and the traditional Chinese medicine decoction pieces comprise part of traditional Chinese medicine slices processed by a production place, original-shape traditional Chinese medicine decoction pieces and decoction pieces processed by cutting and stir-frying. With the development of technology, the traditional Chinese medicine fermented drink as one of the traditional Chinese medicine drinks has unique flavor and is developed quickly, and the lipid-lowering fermented drink is more and more popular with the increase of the number of the existing obese people.
Fermentation is taken as an important step of beverage production, plays an important role in the taste of traditional Chinese medicine beverages, at present, the using amount of lactic acid bacteria is generally obtained according to experience in the fermentation process and is judged, the concentration of traditional Chinese medicine extracting solution, the adding amount of glucose and the adding amount of skim milk powder are difficult to optimize, and the final taste is influenced.
Disclosure of Invention
In order to achieve the above object, the present invention adopts the following technical solutions
On one hand, the invention provides a compound traditional Chinese medicine lipid-lowering fermented beverage, which comprises the following formula components, by weight, 30-100 parts of traditional Chinese medicine extracting solution, 3-5 parts of glucose and 1-3 parts of skim milk powder.
As a preferred technical scheme of the invention, the preparation method of the traditional Chinese medicine extracting solution comprises the steps of weighing 6-8g of lotus leaves, 20-30g of Chinese yam, 6-10g of chrysanthemum, 9-11g of honeysuckle, 4-6g of dried orange peel, 18-22g of cassia seeds, 23-27g of poria cocos and 13-17g of coix seeds, adding a proper amount of water and other auxiliary agents, and decocting the juice to obtain the traditional Chinese medicine extracting solution.
In a second aspect, the invention further provides a research method for the optimal proportion of the compound traditional Chinese medicine lipid-lowering fermented beverage, which specifically comprises the following steps: s1, determining the optimal fermentation amount of the raw material ratio; and S2, determining the optimal fermentation process.
As a preferred technical solution of the present invention, in the step S1, the specific steps of determining the optimal fermentation amount of the raw material ratio are as follows: s11, determining pH value, total acid amount and sensory test items: taking a to-be-detected traditional Chinese medicine beverage sample, and directly detecting the pH value by using a pH meter; accurately measuring 2.0mL of sample solution to be measured, placing the sample solution to be measured in a 150mL conical flask, adding 40-60mL of distilled water, dropwise adding 1-2 drops of phenolphthalein indicator, slowly titrating the sample solution to be measured with 0.2mol/L NaOH titration solution, observing the color of the solution to become red, stopping titration, waiting for 30s, marking that the titration is finished and the color of the solution is not changed any more, recording the volume of consumed NaOH standard solution, and then calculating the total acid amount according to the volume of the NaOH standard solution; evaluating the sense of the sample according to a preset rule; s12, adjusting the pH value of the extracting solution: selecting dilute NaOH solution, and adjusting the pH value of the extracting solution to 6.4-6.6; s13, strain activation: preparing an MRS liquid culture medium, respectively inoculating lactobacillus plantarum and lactobacillus rhamnosus in the liquid culture medium, placing in an incubator at 37 ℃ for standing culture for 24 hours, taking out, selecting proper dilution by adopting a 10-fold gradient dilution method, respectively and uniformly coating a bacterial liquid on a prepared MRS plate, inversely placing in the incubator at 37 ℃ for culture for 36 hours, observing the growth condition of the strain, and selecting a strain with higher growth speed and larger bacterial colony on the plate; s14, determining the most appropriate strain compounding ratio: dividing the traditional Chinese medicine extracting solution into 5 groups, wherein the 1 st group is only inoculated with lactobacillus plantarum, the 2 nd group is only inoculated with lactobacillus rhamnosus, and the 3 rd to 5 th groups are respectively according to the ratio of lactobacillus plantarum: inoculating compound strains in the ratio of lactobacillus rhamnosus to 1:1, 1:1.5 and 1.5:1, wherein the fixed inoculation amount is 4%, placing the strains at 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the lactic acid content in the fermentation liquor at intervals of 4h, and determining the most appropriate strain compound ratio according to the change conditions of the two indexes; s15, determining the most suitable carbon source for the growth of the strain: selecting 4 carbon sources of glucose, fructose, sucrose and lactose, respectively adding the carbon sources into the traditional Chinese medicine extracting solution according to the addition amount of 2%, sterilizing, cooling to 40 ℃, inoculating 4% of mixed strains, wherein the ratio of the strains in the mixed strains is as follows: putting the lactobacillus rhamnosus at the temperature of 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the content of lactic acid in the fermentation liquor at intervals of 4h, and screening out a carbon source favorable for strain growth according to the change conditions of the two indexes; s16, determining the most suitable nitrogen source for the growth of the strain: selecting 3 nitrogen sources of soybean protein isolate, peptone and skim milk powder, respectively adding the nitrogen sources into the traditional Chinese medicine extracting solution by the adding amount of 1%, sterilizing, cooling to 40 ℃, inoculating 4% of mixed strains, wherein the ratio of the strains in the mixed strains is as follows: putting the lactobacillus rhamnosus at the temperature of 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the content of lactic acid in the fermentation liquor at intervals of 4h, and screening out a nitrogen source favorable for strain growth according to the change conditions of the two indexes; s17, obtaining the optimal concentration of the traditional Chinese medicine extracting solution, the optimal addition amount of glucose and the optimal addition amount of the skim milk powder through a single-factor test; s18, on the basis of the step S17, the three-factor three-level orthogonal experiment is carried out on the raw material ratio.
As a preferred technical solution of the present invention, in step S17, the specific steps of determining the optimal concentration of the chinese medicine extract, the optimal amount of glucose added, and the optimal amount of skimmed milk powder added are as follows: determining the optimal concentration of the traditional Chinese medicine extracting solution: respectively selecting the concentration of the traditional Chinese medicine extract from 30%, 40%, 50%, 60% and 70%, the adding amount of glucose is 8%, the adding amount of skim milk powder is 3%, inoculating 4% of compound strain, fermenting at 37 deg.C for 48h, and determining the optimal traditional Chinese medicine liquid concentration according to lactic acid content and sensory score; determining the optimal glucose addition amount: adding 3% of skimmed milk powder into the Chinese medicinal extractive solution, adding glucose in the amount of 0%, 4%, 6%, 8% and 10%, respectively, inoculating 4% of compound strain, fermenting at 37 deg.C for 48 hr, and determining the optimal glucose addition amount according to lactic acid content and sensory score; determining the optimal addition amount of the skim milk powder: adding 8% of glucose into the extracting solution, adding the skim milk powder into the extracting solution according to the adding amounts of 0%, 1%, 2%, 3% and 4%, respectively, inoculating 4% of compound strain, fermenting at 37 ℃ for 48h, and determining the optimal adding amount of the skim milk powder according to the lactic acid content and sensory score.
As a preferred technical solution of the present invention, in step S18, in the orthogonal experiment of the three-factor three-level, the concentrations of the traditional Chinese medicine extract are respectively selected from 30%, 40% and 50%; the addition amounts of glucose are respectively 6%, 8% and 10%; the addition amounts of the skim milk powder are respectively 1%, 2% and 3%.
As a preferred technical solution of the present invention, in the step 2, the specific steps for determining the optimal fermentation process are as follows: s21, determining the optimal lactobacillus inoculation amount: according to the lactobacillus plantarum: adjusting the ratio of lactobacillus rhamnosus to 1:1, adjusting the total inoculation amount to be 1%, 2%, 3%, 4% and 5% to inoculate in the extracting solution, fermenting at 37 ℃ for 48h, measuring the pH value and lactic acid content of the fermentation liquor, and determining the optimal lactobacillus inoculation amount; s22, determining the optimal fermentation temperature: according to the lactobacillus plantarum: inoculating lactobacillus rhamnosus with an inoculum size of 4% to the extractive solution, fermenting at 32 deg.C, 34 deg.C, 36 deg.C, 38 deg.C and 40 deg.C for 48h, measuring pH and lactic acid content of the fermentation broth, and determining the optimal fermentation temperature; s23, determining the optimal fermentation time: according to the lactobacillus plantarum: inoculating lactobacillus rhamnosus with a total inoculation amount of 4% into the extract at a ratio of 1:1, adjusting fermentation time at 37 deg.C, measuring pH value and lactic acid content of the fermentation liquid every 4h, and determining optimal fermentation time; s24, optimizing process parameters: according to the single-factor experiment result center combined design principle, three factors of an inoculation amount A, a fermentation temperature B and a fermentation time C are determined to be 17 experiment points, response surface experiments and analysis are carried out, and fermentation process parameters of the beverage are optimized.
(III) advantageous effects
1. The invention provides a compound traditional Chinese medicine lipid-lowering fermented beverage, which comprises, by weight, 30-100 parts of a traditional Chinese medicine extracting solution, 3-5 parts of glucose and 1-3 parts of skim milk powder, and the preparation method of the traditional Chinese medicine extracting solution comprises the steps of weighing 6-8g of lotus leaves, 20-30g of Chinese yam, 6-10g of chrysanthemum, 9-11g of honeysuckle, 4-6g of dried orange peel, 18-22g of cassia seeds, 23-27g of poria cocos and 13-17g of semen coicis, adding a proper amount of water and other auxiliary agents, and decocting juice to obtain the traditional Chinese medicine extracting solution, wherein the traditional Chinese medicine extracting solution has good taste and a weight-reducing effect;
2. the invention provides a research method for the optimal proportion of a compound traditional Chinese medicine lipid-lowering fermented beverage, which determines that the optimal fermentation strain of the beverage is lactobacillus plantarum by determining the proportion of raw materials: determining that the optimal carbon source is glucose and the optimal nitrogen source is skim milk powder, wherein the ratio of lactobacillus rhamnosus to lactobacillus rhamnosus is 1: 1;
3. the research method of the optimal proportion of the compound traditional Chinese medicine lipid-lowering fermented drink determines the optimal fermentation process, the lactobacillus inoculation amount is 4.5%, the fermentation temperature is 37 ℃, the fermentation time is 50 hours, and the taste of the drink is ensured by determining the optimal proportion and the optimal process.
Drawings
FIG. 1 is a schematic representation of the growth curve of Lactobacillus plantarum in accordance with the invention;
FIG. 2 is a schematic diagram of a growth curve of Lactobacillus rhamnosus according to the present invention;
FIG. 3 is a schematic diagram showing the effect of the ratio of the bacterial strain formulation of the present invention on the lactic acid content of the fermented beverage;
FIG. 4 is a schematic illustration of the effect of the strain formulation ratio of the present invention on the pH of a fermented beverage;
FIG. 5 is a schematic illustration of the effect of the type of carbon source of the present invention on the lactic acid content of a fermented beverage;
FIG. 6 is a schematic diagram showing the effect of the kind of carbon source of the present invention on the pH of fermented beverages;
FIG. 7 is a graph showing the effect of nitrogen source species of the present invention on the lactic acid content of fermented beverages;
FIG. 8 is a schematic representation of the effect of nitrogen source species of the present invention on pH of fermented beverages;
FIG. 9 is a schematic illustration of the effect of concentration of Chinese herbal liquid on lactic acid content of fermented beverages according to the present invention;
FIG. 10 is a graphical representation of the effect of concentration of a liquid Chinese medicine of the present invention on sensory evaluation scores;
FIG. 11 is a graphical representation of the effect of glucose addition on lactic acid content of fermented beverages according to the present invention;
FIG. 12 is a graphical representation of the effect of glucose addition levels on sensory score according to the present invention;
FIG. 13 is a graph showing the effect of the amount of skim milk powder added according to the present invention on the lactic acid content of a fermented beverage;
FIG. 14 is a graph showing the effect of the amount of skim milk powder added according to the present invention on sensory evaluation scores;
FIG. 15 is a graphical representation of the effect of the amount of inoculation of the present invention on the lactic acid content of a fermented beverage;
FIG. 16 is a graph showing the effect of inoculum size of the present invention on pH of fermented beverages;
FIG. 17 is a schematic illustration of the effect of fermentation temperature on lactic acid content of fermented beverages according to the present invention;
FIG. 18 is a schematic of the effect of fermentation temperature on pH of fermented beverages according to the present invention;
FIG. 19 is a schematic of the effect of fermentation time on lactic acid content of fermented beverages according to the present invention;
FIG. 20 is a schematic of the effect of fermentation time on pH of fermented beverages according to the present invention;
FIG. 21 is a line contour plot of the effect of fermentation temperature and inoculum size on lactic acid content of the present invention;
FIG. 22 is a graph of the response of fermentation temperature and inoculum size of the present invention on the effect of lactic acid content;
FIG. 23 is a line contour plot of the effect of inoculum size and fermentation time on lactic acid content of the present invention;
FIG. 24 is a graph showing the effect of inoculum size and fermentation time on lactic acid content in accordance with the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings of the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
A compound traditional Chinese medicine lipid-lowering fermented beverage comprises, by weight, 30-100 parts of traditional Chinese medicine extract, 3-5 parts of glucose and 1-3 parts of skim milk powder.
In the embodiment, the preparation method of the traditional Chinese medicine extracting solution comprises the steps of weighing 6-8g of lotus leaves, 20-30g of Chinese yam, 6-10g of chrysanthemum, 9-11g of honeysuckle, 4-6g of dried orange peel, 18-22g of cassia seeds, 23-27g of poria cocos and 13-17g of semen coicis, adding a proper amount of water and other auxiliary agents, and decocting the juice to obtain the traditional Chinese medicine extracting solution.
As shown in fig. 1 to fig. 24, in a second aspect, the present invention further provides a method for researching an optimal ratio of a compound traditional Chinese medicine lipid-lowering fermented beverage, which specifically includes the following steps:
s1, determining the optimal fermentation amount of the raw material ratio; and S2, determining the optimal fermentation process.
In this embodiment, in step S1, the specific steps of determining the optimal fermentation amount of the raw material ratio are as follows:
s11, determining pH value, total acid amount and sensory test items: taking a to-be-detected traditional Chinese medicine beverage sample, and directly detecting the pH value by using a pH meter; accurately measuring 2.0mL of sample solution to be measured, placing the sample solution to be measured in a 150mL conical flask, adding 40-60mL of distilled water, dropwise adding 1-2 drops of phenolphthalein indicator, slowly titrating the sample solution to be measured with 0.2mol/L NaOH titration solution, observing the color of the solution to become red, stopping titration, waiting for 30s, marking that the titration is finished and the color of the solution is not changed any more, recording the volume of consumed NaOH standard solution, and then calculating the total acid amount according to the volume of the NaOH standard solution; evaluating the sense of the sample according to a preset rule; the content of the sensory evaluation includes color, taste, odor (30 points) and tissue morphology as evaluation indexes, and the specific content is shown in table 1 below:
TABLE 1 sensory evaluation criteria
S12, adjusting the pH value of the extracting solution: selecting dilute NaOH solution, and adjusting the pH value of the extracting solution to 6.4-6.6;
s13, strain activation: preparing an MRS liquid culture medium, respectively inoculating lactobacillus plantarum and lactobacillus rhamnosus in the liquid culture medium, placing in an incubator at 37 ℃ for standing culture for 24 hours, taking out, selecting proper dilution by adopting a 10-fold gradient dilution method, respectively and uniformly coating a bacterial liquid on a prepared MRS plate, inversely placing in the incubator at 37 ℃ for culture for 36 hours, observing the growth condition of the strain, and selecting a strain with higher growth speed and larger bacterial colony on the plate;
it should be noted that, during a specific test, as shown in fig. 1, 0h to 4h are the lag phase of the lactobacillus plantarum, 4h starts to enter the logarithmic growth phase, and starts to stabilize after 24h, the stationary phase ends at 36h, and then enters the decay phase, the strain propagation speed in the logarithmic growth phase is fastest, and after the strain propagation speed enters the stationary phase, the total number of bacteria in the culture medium is the largest, so that the strain in the logarithmic growth phase is selected for inoculation, and therefore, the seed liquid culture time of the lactobacillus plantarum should be controlled to be 4h to 24 h. As shown in figure 2, 0h-4h is the delay phase of the growth of the lactobacillus rhamnosus, 4h begins to enter the logarithmic phase until 20h begins to tend to be stable, the stationary phase is finished at 32h, and then the strain enters the death phase, and the strain in the logarithmic phase is adopted for inoculation, so that the seed liquid culture time of the lactobacillus rhamnosus should be controlled within 4h-20 h. Since the start and end times of the logarithmic growth phase of the two strains are not completely the same, if the two strains are cultured in the seed solution at the same time, the seed solution culture time can be determined to be 8 hours in consideration of experimental feasibility.
S14, determining the most appropriate strain compounding ratio: dividing the traditional Chinese medicine extracting solution into 5 groups, wherein the 1 st group is only inoculated with lactobacillus plantarum, the 2 nd group is only inoculated with lactobacillus rhamnosus, and the 3 rd to 5 th groups are respectively according to the ratio of lactobacillus plantarum: inoculating compound strains in the ratio of lactobacillus rhamnosus to 1:1, 1:1.5 and 1.5:1, wherein the fixed inoculation amount is 4%, placing the strains at 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the lactic acid content in the fermentation liquor at intervals of 4h, and determining the most appropriate strain compound ratio according to the change conditions of the two indexes;
it should be noted that, in a specific experiment, as shown in fig. 3, no matter single-strain fermentation or mixed fermentation of two strains, the lactic acid content in the fermentation liquid shows an increasing trend of different degrees within 48 hours, the lactic acid yield of lactobacillus rhamnosus by single fermentation is higher than that of lactobacillus plantarum by single fermentation, and the lactic acid production capacity of lactobacillus rhamnosus is obviously better than that of lactobacillus plantarum because the lactic acid content can be used for measuring the acid production capacity of lactic acid bacteria. When the two strains are mixed and fermented, the lactic acid yield is obviously higher than that of lactobacillus plantarum when the compound ratio of lactobacillus plantarum to lactobacillus rhamnosus is 1:1, 1:1.5 and 1.5:1, and is slightly higher than that of lactobacillus rhamnosus when the compound ratio of lactobacillus plantarum to lactobacillus rhamnosus is 1:1, the lactic acid yield is the most when the compound ratio of lactobacillus plantarum to lactobacillus rhamnosus is 1:1, and the lactic acid yield is the least when the compound ratio of lactobacillus rhamnosus is 1:1 and 1.5: 1. Therefore, the optimal strain proportion is 1:1, the lactic acid content of the combined strain is changed by 5.67g/kg after fermentation for 48 hours, and the lactic acid content of the combined strain is respectively changed by 1.495g/kg, 4.67g/kg, 5.505g/kg and 5.395g/kg after fermentation for 48 hours, wherein the lactic acid content of the combined strain is only lactobacillus plantarum, only lactobacillus rhamnosus, the compound proportion of lactobacillus plantarum and lactobacillus rhamnosus is 1:1.5, and the compound proportion of lactobacillus plantarum and lactobacillus rhamnosus is 1.5: 1.
As shown in fig. 4, the pH value is continuously decreased with the increase of the fermentation time, which is mainly caused by the continuous accumulation of the generated lactic acid, but the pH values of the strains with different ratios are in a trend of increasing to different degrees within 48h, the strains are more active in the initial stage of fermentation, a large amount of lactic acid is generated, so that the pH value of the fermentation liquid is rapidly decreased, and the growth of the lactic acid bacteria is inhibited by the acidic environment with the continuous decrease of the pH value, so that the pH value is slowly decreased after 24h of fermentation. After fermentation for 48 hours, the pH values of the group of lactobacillus plantarum only, lactobacillus rhamnosus only, lactobacillus plantarum and lactobacillus rhamnosus with the compounding ratio of 1:1.5 and lactobacillus rhamnosus with the compounding ratio of 1.5:1 are respectively reduced to 4.75, 3.88, 3.78, 3.82 and 3.8, namely the pH value of 1:1 is reduced fastest, the pH value of lactobacillus plantarum is reduced slowest, the taste of lactobacillus rhamnosus after fermentation is more stimulated, the lactobacillus plantarum fermentation taste is fine and soft and has obvious aromatic odor generation, so that the quality of the two lactobacillus plantarum compound fermentation product is better.
S15, determining the most suitable carbon source for the growth of the strain: selecting 4 carbon sources of glucose, fructose, sucrose and lactose, respectively adding the carbon sources into the traditional Chinese medicine extracting solution according to the addition amount of 2%, sterilizing, cooling to 40 ℃, inoculating 4% of mixed strains, wherein the ratio of the strains in the mixed strains is as follows: putting the lactobacillus rhamnosus at the temperature of 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the content of lactic acid in the fermentation liquor at intervals of 4h, and screening out a carbon source favorable for strain growth according to the change conditions of the two indexes; it should be noted that, in the specific experiment, the influence of the carbon source type on the lactic acid content of the fermented beverage and the influence of the carbon source type on the pH value of the fermented beverage are obtained, and the specific contents are as follows: lactic acid bacteria can produce acids by fermentation using sugars, but their ability to use sugars varies. As can be seen from fig. 5, the lactic acid bacteria were fermented with four kinds of sugars with increasing fermentation time to produce different amounts of acid, and the consumption of glucose, fructose, sucrose, and lactose were sequentially performed from high to low. The utilization effect of the glucose and the fructose is better than that of the sucrose and the lactose, mainly because the glucose and the fructose are monosaccharides with the simplest structures and can be directly utilized; sucrose and lactose are both classified as disaccharides formed by two molecular monosaccharides, and the disaccharides are utilized only by being hydrolyzed into monosaccharides, and cannot be directly utilized by lactic acid bacteria. After fermentation for 48 hours, the lactic acid content of the glucose, fructose, sucrose and lactose experimental group is 6.275g/kg, 5.54g/kg, 2.485g/kg and 1.885g/kg respectively, and the lactic acid content of the blank group is 1.65 g/kg. From this, it was found that glucose is an appropriate carbon source because glucose has the best utilization effect and the highest lactic acid production amount during fermentation. As shown in fig. 6, lactic acid bacteria produce lactic acid by sugar fermentation, the pH value of the beverage is continuously decreased with the increase of the content of lactic acid, and after 48 hours of fermentation, the pH values of the experimental groups of glucose, fructose, sucrose and lactose are decreased by 3.20, 2.92, 2.39 and 2.03 respectively, and the pH value of the blank group is decreased by 1.73. The acid yield is the most and the pH changes the most when glucose is used as a carbon source. Therefore, glucose was selected as the carbon source.
S16, determining the most suitable nitrogen source for the growth of the strain: selecting 3 nitrogen sources of soybean protein isolate, peptone and skim milk powder, respectively adding the nitrogen sources into the traditional Chinese medicine extracting solution by the adding amount of 1%, sterilizing, cooling to 40 ℃, inoculating 4% of mixed strains, wherein the ratio of the strains in the mixed strains is as follows: putting the lactobacillus rhamnosus at the temperature of 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the content of lactic acid in the fermentation liquor at intervals of 4h, and screening out a nitrogen source favorable for strain growth according to the change conditions of the two indexes; it should be noted that, in the specific experiment, the influence of the nitrogen source type on the lactic acid content of the fermented beverage and the influence of the nitrogen source type on the pH value of the fermented beverage are obtained, and the specific contents are as follows:
lactic acid bacteria cannot synthesize proteins by themselves, but the requirements on nutrient substances are strict, so in order to meet the growth requirements of the lactic acid bacteria, the lactic acid bacteria need to hydrolyze the proteins in the environment to oligopeptides for self utilization. The addition of protein in the beverage can not only increase fragrance and improve taste, but also promote the growth of lactobacillus and improve the quality of the beverage. As shown in FIG. 7, the nitrogen source utilization capacity of the lactic acid bacteria is sequentially from strong to weak peptone, skim milk powder and soybean protein isolate, the fermentation time is 48 hours, the acid production variation is 7.290g/kg, 5.727g/kg and 1.59g/kg, and the blank group is 0.92 g/kg. Lactic acid bacteria produce the most acid using peptone, but fermented beverages have poor odor and taste, so peptone is excluded from the selection of nitrogen sources. The lactic acid bacteria has better utilization effect on the skim milk powder than the soybean protein isolate, which is probably related to the molecular size of the protein, the molecular size of the cow milk protein is 80-120 nm, and the molecular size of the soybean protein is 5000-8000 nm, so the skim milk powder is easier to be utilized. And the skim milk powder is used as a nitrogen source, and the fermented beverage has certain fragrance. Skim milk powder is therefore a suitable source of nitrogen. As shown in FIG. 8, the pH of the beverage was slightly increased by the initial addition of three nitrogen sources, probably due to the buffering action of the protein, and after 48 hours of fermentation, the pH of the peptone, skim milk powder, and soy protein isolate groups decreased by 2.48, 3.27, and 3.13, respectively, and the blank group decreased by 3.02. In consideration of food nutrition and quality, finally, skim milk powder is selected as a nitrogen source.
S17, obtaining the optimal concentration of the traditional Chinese medicine extracting solution, the optimal addition amount of glucose and the optimal addition amount of the skim milk powder through a single-factor test;
s18, on the basis of the step S17, the three-factor three-level orthogonal experiment is carried out on the raw material ratio. In this embodiment, in step S17, the specific steps of determining the optimal concentration of the chinese medicine extract, the optimal amount of glucose to be added, and the optimal amount of skimmed milk powder to be added include: determining the optimal concentration of the traditional Chinese medicine extracting solution: respectively selecting the concentration of the traditional Chinese medicine extract from 30%, 40%, 50%, 60% and 70%, the adding amount of glucose is 8%, the adding amount of skim milk powder is 3%, inoculating 4% of compound strain, fermenting at 37 deg.C for 48h, and determining the optimal traditional Chinese medicine liquid concentration according to lactic acid content and sensory score; in a specific experiment, as shown in fig. 9, after fermentation for 48 hours, the content of lactic acid in the experimental group of the traditional Chinese medicine liquid with the concentration of 30% -70% was respectively changed by 8.35g/kg, 8.52g/kg, 8.56g/kg, 8.41g/kg and 8.33 g/kg. The concentration of the traditional Chinese medicine liquid has less influence on the content of the lactic acid probably because the traditional Chinese medicine liquid contains less nutrient substances available for the lactic acid bacteria, and the lactic acid in the beverage is mainly generated because the lactic acid bacteria utilize the saccharides and proteins provided by the carbon source and the nitrogen source. When the concentration of the Chinese medicinal liquid reaches 40%, the content of lactic acid is almost not changed, the concentration of the Chinese medicinal liquid is continuously increased, the content of lactic acid is slightly reduced, and the content of lactic acid is reduced probably because the growth substances of lactic acid bacteria, such as organic acid and the like, in the Chinese medicinal liquid are increased, and the environment is not beneficial to the growth of the lactic acid bacteria.
The mixing ratio of the traditional Chinese medicine extract and water is different, and the smell, taste and color of the fermented beverage are also different. As can be seen from fig. 10, as the concentration of the Chinese medicinal liquid increases, the sensory score of the beverage increases first and then decreases, the beverage fermented by the Chinese medicinal liquid concentration of 30% has thinner mouthfeel and lighter color and lacks Chinese medicinal and fermented aroma, the beverage fermented by the Chinese medicinal liquid concentrations of 60% and 70% has sour and astringent mouthfeel and lighter fermented aroma, and the production cost is correspondingly increased. Therefore, the concentration of the traditional Chinese medicine liquid is preferably 30-50%.
Determining the optimal glucose addition amount: adding 3% of skimmed milk powder into the Chinese medicinal extractive solution, adding glucose in the amount of 0%, 4%, 6%, 8% and 10%, respectively, inoculating 4% of compound strain, fermenting at 37 deg.C for 48 hr, and determining the optimal glucose addition amount according to lactic acid content and sensory score; in the specific experiment, as can be seen from FIG. 11, the lactic acid content increased with the increase in the amount of glucose added, and after 48 hours of fermentation, the lactic acid contents of the 4% to 10% experimental groups were 7.4g/kg, 8.0g/kg, 8.7g/kg, and 8.9g/kg, respectively. It can be seen that the growth of lactic acid bacteria can be promoted by low concentration of glucose, and when 10% of glucose is added, the content of lactic acid is increased by only 0.22g/kg compared with the content of lactic acid in the group with 8% of glucose, probably because the osmotic pressure of lactic acid bacteria cells is changed by higher concentration of glucose. As can be seen from fig. 12, the addition amount of glucose has a large influence on the sensory evaluation of the beverage, the glucose can improve the bitter taste of the Chinese medicinal liquid and can also promote lactic acid bacteria to produce lactic acid, the taste of the beverage is optimal only when the sugar-acid ratio of the beverage is coordinated, and the taste of the fermented beverage is slightly sour when the addition amount of glucose is less than 8%; when the addition amount of the glucose is 8%, the fermented beverage is moderate in sweetness and sourness; when the addition amount of glucose is continuously increased, the glucose in the fermented beverage is not completely utilized, and a large amount of glucose is accumulated to cause a sweet taste. Therefore, a preferred level of glucose addition is 6% to 10%.
Determining the optimal addition amount of the skim milk powder: adding 8% of glucose into the extracting solution, adding the skim milk powder into the extracting solution according to the adding amounts of 0%, 1%, 2%, 3% and 4%, respectively, inoculating 4% of compound strain, fermenting at 37 ℃ for 48h, and determining the optimal adding amount of the skim milk powder according to the lactic acid content and sensory score. In the concrete test, as shown in fig. 13, the influence of the addition amount of the skim milk powder on the lactic acid content was large, and the lactic acid content of the test group to which 1% to 4% skim milk powder was added after 48 hours of fermentation was 6.3g/kg, 7.2g/kg, 8.51g/kg, and 8.782g/kg, respectively. It can be seen that the lactic acid content increases with the amount of the skim milk powder added. As can be seen from fig. 14, when the added amount of skim milk powder was 1%, the amount of protein available for lactic acid bacteria was small, fermentation was incomplete, the amount of lactic acid produced was small, and the sour taste of the fermented beverage was insufficient; when the addition amount of the skim milk powder is 2% or 3%, the sweet-sour ratio of the fermented beverage is coordinated, and the taste is moderate; when the addition amount of the skim milk powder is 4%, the acid production amount is large, the fermented beverage tastes sour, and loses luster and the quality is poor. Therefore, the preferable level of the addition amount of the skim milk powder is 1 to 3 percent.
In this embodiment, in step S18, in the orthogonal experiment of the three-factor three-level, the concentrations of the chinese medicine extract are respectively 30%, 40% and 50%; the addition amounts of glucose are respectively 6%, 8% and 10%; the addition amounts of the skim milk powder are respectively 1%, 2% and 3%. It should be noted that, three-factor three-level orthogonal experiments are performed on the raw material ratio, and as can be seen from table 2, in terms of the content of lactic acid, the influence of each factor on the content of lactic acid is ranked as follows: c (the added amount of the skim milk powder) is more than B (the added amount of the glucose) is more than A (the concentration of the Chinese medicinal liquid), and the optimal combination is A3B3C 3; in the aspect of sensory score, the influence of each factor on the sensory score is ranked as follows: the optimal combination is A2B2C3, wherein A (traditional Chinese medicine liquid concentration) > B (glucose addition quantity) > C (skim milk powder addition quantity). In order to save cost while ensuring the taste, the A2B2C3 combination scheme is selected for comprehensive consideration. Therefore, the concentration of the traditional Chinese medicine liquid of 40%, the addition amount of glucose of 8% and the addition amount of skim milk powder of 3% are selected, and at this time, the lactic acid content of the fermented beverage is 8.62g/kg, and the sensory score is 96.
Table 2 orthogonal experimental results of raw material ratio table
As can be seen from Table 3, the amount of glucose added (B) had a significant effect on the lactic acid content (P < 0.05), the amount of skim milk powder added (C) had a very significant effect on the lactic acid content (P < 0.01), and both the concentration of the herbal solution and the amount of glucose added had significant effects on the sensory score (P < 0.05).
TABLE 3 ANOVA RESULT TABLE
In table 3, differences were very significant (P < 0.01); marked variability (P < 0.05); - -was not significantly different (P > 0.05).
In conclusion, through single-factor experiments and orthogonal experimental analysis, the raw material ratio of the beverage is optimized, and the optimal raw material ratio is finally determined as follows: the concentration of the traditional Chinese medicine liquid is 40%, the adding amount of glucose is 8%, and the adding amount of the skim milk powder is 3%.
In this embodiment, in the step 2, the specific steps for determining the optimal fermentation process are as follows:
s21, determining the optimal lactobacillus inoculation amount: according to the lactobacillus plantarum: adjusting the ratio of lactobacillus rhamnosus to 1:1, adjusting the total inoculation amount to be 1%, 2%, 3%, 4% and 5% to inoculate in the extracting solution, fermenting at 37 ℃ for 48h, measuring the pH value and lactic acid content of the fermentation liquor, and determining the optimal lactobacillus inoculation amount; in the specific experiment, as can be seen from fig. 15 and 16, the lactic acid contents of the experimental groups with 1% -5% inoculum sizes after 48 hours of fermentation were 8.485g/kg, 8.775g/kg, 9.1g/kg and 8.97g/kg, respectively; the pH values were 3.62, 3.6, 3.58, 3.55, 3.57. The lactic acid content of the experimental group with the inoculation amount of 4 percent is increased by 0.325g/kg compared with the lactic acid content of the experimental group with the inoculation amount of 3 percent, and the lactic acid content of the experimental group with the inoculation amount of 5 percent is reduced by 0.12g/kg compared with the lactic acid content of the experimental group with the inoculation amount of 4 percent. Therefore, an inoculum size of 3% -5% was selected as the level of response surface experiments.
S22, determining the optimal fermentation temperature: according to the lactobacillus plantarum: inoculating lactobacillus rhamnosus with an inoculum size of 4% to the extractive solution, fermenting at 32 deg.C, 34 deg.C, 36 deg.C, 38 deg.C and 40 deg.C for 48h, measuring pH and lactic acid content of the fermentation broth, and determining the optimal fermentation temperature; it should be noted that in this experiment, the temperature plays an important role in the growth and metabolism of lactic acid bacteria during the whole fermentation process of the beverage, and affects the quality of the fermented beverage. As can be seen from FIGS. 17 and 18, after 48 hours of fermentation, the lactic acid contents of the experimental groups at the fermentation temperature of 32 ℃ to 40 ℃ are respectively 8.15g/kg, 8.33g/kg, 9.02g/kg, 8.98g/kg and 8.65 g/kg; the pH values were 3.66, 3.58, 3.52, 3.53, 3.56. Therefore, the fermentation temperature is too low, the initial activation time of the strain is longer, the subsequent propagation speed is slower, and the yield is lower; when the fermentation temperature is too high, the initial strain grows rapidly, the pH value is rapidly reduced along with the generation of lactic acid, but the lactic acid bacteria can inhibit the activity of the lactic acid bacteria when entering an acidic environment rapidly, so that the lactic acid bacteria can enter an apoptosis stage rapidly. Too low and too high temperature are not beneficial to the growth of lactic acid bacteria and affect the quality of fermented beverage. Therefore, a fermentation temperature of 35 ℃ to 39 ℃ was selected as the level of response surface experiments.
S23, determining the optimal fermentation time: according to the lactobacillus plantarum: inoculating lactobacillus rhamnosus with a total inoculation amount of 4% into the extract at a ratio of 1:1, adjusting fermentation time at 37 deg.C, measuring pH value and lactic acid content of the fermentation liquid every 4h, and determining optimal fermentation time;
it should be noted that, in the test process, the influence of the fermentation time on the lactic acid content and the pH value of the fermented beverage is shown in fig. 19 and fig. 20, the lactic acid content increases with the increase of the fermentation time, the pH decreases with the increase of the fermentation time, the rates of the increase of the lactic acid content and the decrease of the pH value are both fast first and slow second in the whole process, the fermentation time reaches 48h, the change curve of the lactic acid content and the pH value tends to be smooth, the lactic acid content increases by 0.21 when the fermentation time is 48h compared with 44h, and the pH value decreases by 0.04; the lactic acid content is only increased by 0.08 when the time is 52 hours compared with 48 hours, and the pH value is only reduced by 0.01 and basically has no change. Therefore, fermentation times of 44h to 48h were selected as the level of response surface experiments.
S24, optimizing process parameters: according to the single-factor experiment result center combined design principle, three factors of an inoculation amount A, a fermentation temperature B and a fermentation time C are determined to be 17 experiment points, response surface experiments and analysis are carried out, and fermentation process parameters of the beverage are optimized. It should be noted that, in the testing process, according to the single-factor experimental result and the Box-Behnken center combination design principle, the factor level table is determined, and the response surface experiment is performed, and the results are shown in tables 4 and 5:
table 4 level table of experimental factors of response surface
Table 5 table of experimental results of response surface
Regression fitting is carried out on the data obtained in the table 5 by using Design-expert8.0.6 software, and a multivariate quadratic equation of the lactic acid content (R1) to the seed amount (A), the fermentation temperature (B) and the fermentation time (C) is obtained as follows:
Y＝9.06+0.14A+0.15B+0.08C+0.15AB+0.18AC-0.023BC-0.27A2-0.44B2-0.19C2
as can be seen from the observation of Table 6, the interaction between the interaction terms AB and AC has a very significant effect on the lactic acid content (P < 0.01); the interaction of the interaction term BC had no significant effect on lactate content (P > 0.05), indicating a non-simple linear relationship between the selected factors and the response values. P of the regression model is less than 0.0001, which indicates that the regression model has high significance; meanwhile, the P of the missimulation error term is 0.0780 & gt 0.05, which shows that the simulation has higher reliability. The regression model with the coefficient of determination R2 equal to 0.9958 and the correction coefficient of determination Radj2 equal to 0.9831 demonstrated that the response (lactic acid content in R1) changed by 98.31% from the model, indicating that the model fit better.
As can be seen from the F value items in Table 6, the influence of A, B, C factors on the content of lactic acid is in the order of magnitude: fermentation temperature (B) > inoculation amount (A) > fermentation time (C).
TABLE 6 regression equation ANOVA
From the results of Table 6, the contour plot and the response surface 3D plot were plotted using Design-Expert8.0.6 software, and the influence of various factors on the lactic acid content was observed, as shown in FIGS. 21 to 24.
From the observation of FIGS. 21 to 24, it can be seen that the contour plots between the fermentation temperature (B) and the inoculation amount (A) and between the inoculation amount (A) and the fermentation time (C) are both elliptical, indicating that both sets of interactions are significant, wherein the response surface of the fermentation temperature graph has a steeper change surface than that of the inoculation amount, indicating that the lactic acid content is more sensitive to the change of the fermentation temperature; the change surface of the response surface graph of the inoculation amount is steeper than that of the fermentation time, which shows that the lactic acid content is more sensitive to the change of the inoculation amount, and the result is consistent with the result of the anova.
The optimal fermentation process of the fermented beverage obtained by the analysis of Desgin-Expert8.0.6 software is as follows: the inoculation amount is 4.47%, the fermentation temperature is 37.49 ℃, and the fermentation time is 49.63h, so that the lactic acid content can reach 9.12638g/kg under the condition. For the operability of the experiment, the process was adjusted to an inoculum size of 4.5%, a fermentation temperature of 37 ℃ and a fermentation time of 50 h. Meanwhile, in order to further determine the accuracy of the response surface method, the process is verified, and as shown in Table 7, when the inoculation amount is 4.5%, the fermentation temperature is 37 ℃, and the fermentation time is 50 hours, the lactic acid content can reach 9.09g/kg under the condition. The actual measured value is close to the predicted value of the software, and the response surface method is proved to be capable of effectively optimizing the beverage fermentation process parameters.
TABLE 7 lactic acid content comparison table for optimal fermentation process
In conclusion, the fermentation process conditions of the beverage are optimized, response surface design is carried out on the basis of single-factor experiments, and the optimal fermentation process parameters are obtained as follows: the inoculation amount is 4.5 percent, the fermentation temperature is 37 ℃, and the fermentation time is 50 hours. Under the condition, the content of the lactic acid can reach 9.09 g/kg.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. The compound traditional Chinese medicine lipid-lowering fermented drink is characterized by comprising, by weight, 30-100 parts of traditional Chinese medicine extract, 3-5 parts of glucose and 1-3 parts of skim milk powder.
2. The compound traditional Chinese medicine lipid-lowering fermented drink according to claim 2, which is characterized in that: the preparation method of the traditional Chinese medicine extracting solution comprises the steps of weighing 6-8g of lotus leaves, 20-30g of Chinese yam, 6-10g of chrysanthemum, 9-11g of honeysuckle, 4-6g of dried orange peel, 18-22g of cassia seeds, 23-27g of poria cocos and 13-17g of coix seeds, adding a proper amount of water and other auxiliary agents, and decocting the juice to obtain the traditional Chinese medicine extracting solution.
3. A research method for the optimal proportion of a compound traditional Chinese medicine lipid-lowering fermented drink is characterized by comprising the following steps: the method specifically comprises the following steps:
s1, determining the optimal fermentation amount of the raw material ratio;
and S2, determining the optimal fermentation process.
4. The method for researching the optimal proportion of the compound traditional Chinese medicine lipid-lowering fermented drink according to claim 3, is characterized in that: in step S1, the specific steps of determining the optimal fermentation amount of the raw material ratio are as follows:
s11, determining pH value, total acid amount and sensory test items: taking a to-be-detected traditional Chinese medicine beverage sample, and directly detecting the pH value by using a pH meter; accurately measuring 2.0mL of sample solution to be measured, placing the sample solution to be measured in a 150mL conical flask, adding 40-60mL of distilled water, dropwise adding 1-2 drops of phenolphthalein indicator, slowly titrating the sample solution to be measured with 0.2mol/L NaOH titration solution, observing the color of the solution to become red, stopping titration, waiting for 30s, marking that the titration is finished and the color of the solution is not changed any more, recording the volume of consumed NaOH standard solution, and then calculating the total acid amount according to the volume of the NaOH standard solution; evaluating the sense of the sample according to a preset rule;
s12, adjusting the pH value of the extracting solution: selecting dilute NaOH solution, and adjusting the pH value of the extracting solution to 6.4-6.6;
s13, strain activation: preparing an MRS liquid culture medium, respectively inoculating lactobacillus plantarum and lactobacillus rhamnosus in the liquid culture medium, placing in an incubator at 37 ℃ for standing culture for 24 hours, taking out, selecting proper dilution by adopting a 10-fold gradient dilution method, respectively and uniformly coating a bacterial liquid on a prepared MRS plate, inversely placing in the incubator at 37 ℃ for culture for 36 hours, observing the growth condition of the strain, and selecting a strain with higher growth speed and larger bacterial colony on the plate;
s14, determining the most appropriate strain compounding ratio: dividing the traditional Chinese medicine extracting solution into 5 groups, wherein the 1 st group is only inoculated with lactobacillus plantarum, the 2 nd group is only inoculated with lactobacillus rhamnosus, and the 3 rd to 5 th groups are respectively according to the ratio of lactobacillus plantarum: inoculating compound strains in the ratio of lactobacillus rhamnosus to 1:1, 1:1.5 and 1.5:1, wherein the fixed inoculation amount is 4%, placing the strains at 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the lactic acid content in the fermentation liquor at intervals of 4h, and determining the most appropriate strain compound ratio according to the change conditions of the two indexes;
s15, determining the most suitable carbon source for the growth of the strain: selecting 4 carbon sources of glucose, fructose, sucrose and lactose, respectively adding the carbon sources into the traditional Chinese medicine extracting solution according to the addition amount of 2%, sterilizing, cooling to 40 ℃, inoculating 4% of mixed strains, wherein the ratio of the strains in the mixed strains is as follows: putting the lactobacillus rhamnosus at the temperature of 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the content of lactic acid in the fermentation liquor at intervals of 4h, and screening out a carbon source favorable for strain growth according to the change conditions of the two indexes;
s16, determining the most suitable nitrogen source for the growth of the strain: selecting 3 nitrogen sources of soybean protein isolate, peptone and skim milk powder, respectively adding the nitrogen sources into the traditional Chinese medicine extracting solution by the adding amount of 1%, sterilizing, cooling to 40 ℃, inoculating 4% of mixed strains, wherein the ratio of the strains in the mixed strains is as follows: putting the lactobacillus rhamnosus at the temperature of 37 ℃ for fermentation for 48h, measuring the pH value of the fermentation liquor and the content of lactic acid in the fermentation liquor at intervals of 4h, and screening out a nitrogen source favorable for strain growth according to the change conditions of the two indexes;
s17, obtaining the optimal concentration of the traditional Chinese medicine extracting solution, the optimal addition amount of glucose and the optimal addition amount of the skim milk powder through a single-factor test;
s18, on the basis of the step S17, the three-factor three-level orthogonal experiment is carried out on the raw material ratio.
5. The method for researching the optimal proportion of the compound traditional Chinese medicine lipid-lowering fermented drink according to claim 4, is characterized in that: in step S17, the specific steps of determining the optimal concentration of the chinese medicine extract, the optimal amount of glucose added, and the optimal amount of skimmed milk powder added are:
determining the optimal concentration of the traditional Chinese medicine extracting solution: respectively selecting the concentration of the traditional Chinese medicine extract from 30%, 40%, 50%, 60% and 70%, the adding amount of glucose is 8%, the adding amount of skim milk powder is 3%, inoculating 4% of compound strain, fermenting at 37 deg.C for 48h, and determining the optimal traditional Chinese medicine liquid concentration according to lactic acid content and sensory score;
determining the optimal glucose addition amount: adding 3% of skimmed milk powder into the Chinese medicinal extractive solution, adding glucose in the amount of 0%, 4%, 6%, 8% and 10%, respectively, inoculating 4% of compound strain, fermenting at 37 deg.C for 48 hr, and determining the optimal glucose addition amount according to lactic acid content and sensory score;
determining the optimal addition amount of the skim milk powder: adding 8% of glucose into the extracting solution, adding the skim milk powder into the extracting solution according to the adding amounts of 0%, 1%, 2%, 3% and 4%, respectively, inoculating 4% of compound strain, fermenting at 37 ℃ for 48h, and determining the optimal adding amount of the skim milk powder according to the lactic acid content and sensory score.
6. The method for researching the optimal proportion of the compound traditional Chinese medicine lipid-lowering fermented drink according to claim 5, is characterized in that: in the step S18, in the orthogonal experiment of the three-factor three-level, the concentrations of the traditional Chinese medicine extracting solution are respectively selected to be 30%, 40% and 50%; the addition amounts of glucose are respectively 6%, 8% and 10%; the addition amounts of the skim milk powder are respectively 1%, 2% and 3%.
7. The method for researching the optimal proportion of the compound traditional Chinese medicine lipid-lowering fermented drink according to claim 3, is characterized in that: in the step 2, the specific steps for determining the optimal fermentation process are as follows:
s21, determining the optimal lactobacillus inoculation amount: according to the lactobacillus plantarum: adjusting the ratio of lactobacillus rhamnosus to 1:1, adjusting the total inoculation amount to be 1%, 2%, 3%, 4% and 5% to inoculate in the extracting solution, fermenting at 37 ℃ for 48h, measuring the pH value and lactic acid content of the fermentation liquor, and determining the optimal lactobacillus inoculation amount;
s22, determining the optimal fermentation temperature: according to the lactobacillus plantarum: inoculating lactobacillus rhamnosus with an inoculum size of 4% to the extractive solution, fermenting at 32 deg.C, 34 deg.C, 36 deg.C, 38 deg.C and 40 deg.C for 48h, measuring pH and lactic acid content of the fermentation broth, and determining the optimal fermentation temperature;
s23, determining the optimal fermentation time: according to the lactobacillus plantarum: inoculating lactobacillus rhamnosus with a total inoculation amount of 4% into the extract at a ratio of 1:1, adjusting fermentation time at 37 deg.C, measuring pH value and lactic acid content of the fermentation liquid every 4h, and determining optimal fermentation time;
s24, optimizing process parameters: according to the single-factor experiment result center combined design principle, three factors of an inoculation amount A, a fermentation temperature B and a fermentation time C are determined to be 17 experiment points, response surface experiments and analysis are carried out, and fermentation process parameters of the beverage are optimized.
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