CN103798703A - Preparation method capable of optimizing fermented vegetable juice through response surface methodology - Google Patents
Preparation method capable of optimizing fermented vegetable juice through response surface methodology Download PDFInfo
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- 235000021121 fermented vegetables Nutrition 0.000 title abstract 3
- 238000010993 response surface methodology Methods 0.000 title abstract 3
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 24
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 18
- 238000012360 testing method Methods 0.000 claims description 17
- 239000002253 acid Substances 0.000 claims description 16
- 239000002054 inoculum Substances 0.000 claims description 16
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
- A23L19/20—Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L23/00—Soups; Sauces; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
- A23L7/104—Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms
-
- 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
- A23V2400/121—Brevis
-
- 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
- A23V2400/143—Fermentum
-
- 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
-
- 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
- A23V2400/31—Leuconostoc
- A23V2400/325—Pseudomesenteroides
Abstract
The invention relates to a preparation method capable of optimizing fermented vegetable juice through response surface methodology. The method comprises the following steps: 1, preparing vegetables, namely washing celery and lotus cabbages, cutting the vegetables into segments, and putting the vegetables into a tank according to an equal mass ratio; 2, preparing noodle soup, namely respectively preparing flour and water into the noodle soup according to different horizontal gradients; 3, respectively adding the noodle soup and mixed bacteria into the tank filled with the vegetables for fermenting according to different horizontal gradients, thus obtaining the fermented vegetable juice which corresponds to different inoculation amounts and different ratios of flour to water and vegetables to noodle soup; 4, performing experimental design and statistical analysis, namely (1) performing a single factor experiment, and (2) performing response surface methodology design, namely establishing a quadric multiple regression equation based on three factors including the inoculation amounts, the ratio of flour to water and the ratio of vegetables to noodle soup according to the result of the single factor experiment; and 5, analyzing the experimental result and optimizing, namely performing drawing analysis by Design Expert 8.0 software, thus obtaining a response surface of the regression equation, a contour map of the response surface and the like. The method is simple in operation and suitable for industrial production.
Description
Technical field
The present invention relates to food and biological technical field, relate in particular to a kind of preparation method of response surface method optimization pulp-water.
Background technology
Pulp-water is the fermented food that NORTHWEST CHINA area has characteristic, be generally by after the first blanching of raw material (celery or bitter maror etc.), then put into hot water, with old pulp-water be introduction, then add noodle soup until fermenting-ripening.The pulp-water soup juice of fermenting-ripening is slightly creamy white, smell delicate fragrance, mouthfeel acid alcohol.Pulp-water can directly be scooped out and add white sugar to drink, and its soup juice sweet and sour taste adds green onion and the Chinese prickly ash of boiling in water for a while, then dress with soy, vinegar, etc. oil, then adds noodles to make pulp-water face after also can boiling.The Northwest summer is hot dry, and edible pulp-water or pulp-water face clearly go greasy, have the effects such as fat-reducing, appetizing, is the food of first-selected clearing away heat and removing summer.
Along with the raising of people's living standard, the food of health, health, nutrition more and more receives publicity.Dominant microflora in pulp-water is lactic acid bacteria, and its adjustable human body intestinal canal function improves immunity, suppresses tumor growth, and produced simultaneously lactic acid is not only the main component of pulp-water tart flavour, also can reduce the pH value of pulp-water, avoids other living contaminants.In pulp-water, also contain the Determination of Organic Acids such as tartaric acid, acetic acid, have antibacterial, antiviral, suppress the generation of cerebrum lipid peroxide, anti-inflammatory, anti-sudden change and the effect such as anticancer.
What current domestic research pulp-water was more is separation and the evaluation of wherein microorganism, and has obtained certain achievement in research.Present stage, is mainly directly putting type fermented method for the Study on Preparation of pulp-water, and its emphasis technological process is: clean-segment-blanching 2 ~ 5min of dish enters in proportion tank-add noodle soup-access bacterial classification-25 ~ 30 ℃ and ferments 5 ~ 7 days.Because of each department food custom difference, the kind of the dish of access and component, the sticky degree of noodle soup are also different, and the quality of the pulp-water of fermenting-ripening gives a mark to evaluate its quality mainly with the organoleptic indicator such as color and luster, mouthfeel, without effective convincingness.Patent CN201210470488.5 method of making pulp-water dish, it focuses on making the zymotechnique of pulp-water dish and the elaboration of parameter, only determined the ratio of access bacterial classification by single factor analysis, the ratio of water and flour, the factor such as fermentation temperature, time, zymotechnique is not carried out preferably, the nutritional properties of pulp-water not being described yet.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of simple to operate, preparation method that the response surface method that is easy to suitability for industrialized production is optimized pulp-water.
For addressing the above problem, a kind of response surface method of the present invention is optimized the preparation method of pulp-water, comprises the following steps:
(1) vegetables preparation: segment after respectively celery, lotus flower dish being cleaned, and in etc. the ratio of quality put into tank;
(2) noodle soup preparation: the ratio that the water that is 90 ~ 100 ℃ by flour and temperature is 1kg:60mL, 1 kg:80 mL, 1 kg:100 mL in horizontal gradient is respectively mixed with noodle soup;
(3) in the tank that is placed with described vegetables, be 1 kg:20 mL by horizontal gradient respectively, 1 kg:30 mL, the ratio of 1 kg:40 mL adds described noodle soup, and the inoculum concentration that is 5%, 7%, 9% by horizontal gradient respectively accesses Mixed Microbes, be the condition bottom fermentation 4 ~ 6 days of 20 ~ 40 ℃ in temperature, can obtain the pulp-water corresponding to the ratio of different vaccination amount, flour/water and vegetables/noodle soup;
(4) experimental design and statistical analysis:
1. single factor experiment:
Change successively the ratio of inoculum concentration, flour/water and vegetables/noodle soup, take DPPH clearance rate, viable count, total acid as testing index, then calculate according to the following formula its overall target:
Wherein: viable count is measured and adopted viable count in dilution-plate method working sample, and dilution factor is 10
-6, 10
-8; Each dilution factor does 3 repetitions; Above-mentioned flat board is cultivated to 24h at 30 ℃ of constant incubators, number total plate count;
Acidity assaying:
Take 25g test solution, be placed in 250ml triangular flask, add the phenolphthalein indicator of 40ml water and 0.2ml1%; Be titrated to blush 30s with 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide and do not fade, record consumes the numerical value of the volume of 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide, is denoted as V
1, same sample is measured twice; Blank test: water replaces test solution, by above-mentioned steps operation, record consumes the numerical value of the volume of 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide, is denoted as V
2; Then be calculated as follows total acid:
In formula:
X---in food the mass fraction of total acid in X, numerical value with grams per kilogram, g/kg (or g/L) represents;
The concentration of C---Standard Volumetric Solutions for Sodium Hydroxide, mol/L:
V
1---when titration test solution, consume the volume of Standard Volumetric Solutions for Sodium Hydroxide, mL;
V
2---when blank test, consume the volume of Standard Volumetric Solutions for Sodium Hydroxide, mL;
The extension rate of F---test solution;
K---the conversion coefficient of acid; Wherein the conversion coefficient of various acid is respectively: malic acid 0.067; Acetic acid 0.060; Tartaric acid 0.075; Citric acid 0.064; Containing the citric acid 0.070 () of a part crystallization water; Lactic acid 0.090; Hydrochloric acid 0.036; Phosphoric acid 0.049;
The quality of m---sample, g;
The mensuration of DPPH clearance rate:
First precision takes DPPH standard items 8mg, is placed in 100 mL volumetric flasks, and with methyl alcohol dissolving constant volume, obtaining concentration is 2 × 10
-4the DPPH standard reserving solution of mol/L, at 4 ~ 10 ℃, low temperature keeps in Dark Place for subsequent use; Then, after described storing solution being mixed in the ratio of 3mL:5mL with methyl alcohol, obtain working solution; Secondly, after methyl alcohol is mixed in the ratio of 1mL:3mL with described working solution, utilize UV-VIS spectrophotometry to measure absorbance in 517 nm places, be designated as A
blank=0.681; Finally, after the pulp-water of fermenting-ripening is mixed in the ratio of 1mL:3mL with methyl alcohol, get 10mL pulp-water centrifugal 10 min under 8000 r/min, obtain supernatant, after this supernatant mixes in the ratio of 1mL:3mL with described working solution, reacting at normal temperature without light 20 min, survey absorbance in 517 nm places and are designated as A
sample, parallel determination 2 times, calculate as follows DPPH clearance rate:
2. response surface method optimal design:
According to single factor experiment result, with these 3 factors of ratio of inoculum concentration, flour/water and vegetables/noodle soup, utilize Design Expert 8.0 softwares to carry out experimental design according to Box-Behnken design principle, with flour/water X
1, vegetables/noodle soup X
2with inoculum concentration X
3for independent variable, take overall target as response y, set up polynary quadratic regression equation:
Y=69.08+3.30X
1+11.27?X
2+9.99?X
3+3.92?X
1?X
2+5.93?X
1?X
3-2.31?X
2?X
3-9.18?X
1 2-4.43?X
2 2-8.44?X
3 2；
(5) interpretation and optimization:
Utilize Design Expert 8.0 softwares according to the analysis of drawing of polynary quadratic regression equation, obtain response surface and the contour map thereof of regression equation.
Described Mixed Microbes refers to that Lactobacillus brevis (Lactobacillus brevis), Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus fermenti (Lactobacillus fermentum), leuconostoc pseudomesenteroides (Lactobacillus pseudomesenteroides) are obtained by mixing in the ratio of 1g:1g:1g:1g, described Lactobacillus brevis (Lactobacillus brevis) (deposit number is CGMCC NO:6239), Lactobacillus plantarum (Lactobacillus plantarum) (deposit number is CGMCC NO:20242), lactobacillus fermenti (Lactobacillus fermentum) (deposit number is CGMCC NO:6233), leuconostoc pseudomesenteroides (Lactobacillus pseudomesenteroides) (deposit number is CGMCC NO:22568) is all from Chinese common micro-organisms culture presevation administrative center (depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences).
The present invention compared with prior art has the following advantages:
1, compared with Orthogonal Method, the response surface analysis method that the present invention adopts Box-Behnken Designs (BBD) center combination to design a model, just can draw optimum results by 3 changed factors, 3 levels and a small amount of experimental group (only 17 groups of experiments), obtain nutritiously, color and luster is limpid, the pulp-water of mouthfeel delicate fragrance.
2, the present invention is simple to operate, practical, for the efficient utilization of pulp-water suitability for industrialized production provides foundation.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 is the response surface figure of vegetables/noodle soup of the present invention and two impacts of factor reciprocation on overall target of flour/water.
Fig. 2 is the contour map of vegetables/noodle soup of the present invention and two impacts of factor reciprocation on overall target of flour/water.
Fig. 3 is the response surface figure of inoculum concentration of the present invention and two impacts of factor reciprocation on overall target of flour/water.
Fig. 4 is the contour map of inoculum concentration of the present invention and two impacts of factor reciprocation on overall target of flour/water.
Fig. 5 is the response surface figure of inoculum concentration of the present invention and two impacts of factor reciprocation on overall target of vegetables/noodle soup.
Fig. 6 is the contour map of inoculum concentration of the present invention and two impacts of factor reciprocation on overall target of vegetables/noodle soup.
The specific embodiment
Response surface method is optimized a preparation method for pulp-water, comprises the following steps:
(1) vegetables preparation: segment after respectively celery, lotus flower dish being cleaned, and in etc. the ratio of quality put into tank.
(2) noodle soup preparation: the ratio that the water that is 90 ~ 100 ℃ by flour and temperature is 1kg:60mL, 1 kg:80 mL, 1 kg:100 mL in horizontal gradient is respectively mixed with noodle soup.
(3) in the tank that is placed with vegetables, be 1 kg:20 mL by horizontal gradient respectively, 1 kg:30 mL, the ratio of 1 kg:40 mL adds noodle soup, and the inoculum concentration that is 5%, 7%, 9% by horizontal gradient respectively accesses Mixed Microbes, be the condition bottom fermentation 4 ~ 6 days of 20 ~ 40 ℃ in temperature, can obtain the pulp-water corresponding to the ratio of different vaccination amount, flour/water and vegetables/noodle soup.
Wherein: Mixed Microbes refers to that Lactobacillus brevis (Lactobacillus brevis), Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus fermenti (Lactobacillus fermentum), leuconostoc pseudomesenteroides (Lactobacillus pseudomesenteroides) are obtained by mixing in the ratio of 1g:1g:1g:1g, Lactobacillus brevis (Lactobacillus brevis) (deposit number is CGMCC NO:6239), Lactobacillus plantarum (Lactobacillus plantarum) (deposit number is CGMCC NO:20242), lactobacillus fermenti (Lactobacillus fermentum) (deposit number is CGMCC NO:6233), leuconostoc pseudomesenteroides (Lactobacillus pseudomesenteroides) (deposit number is CGMCC NO:22568) is all from Chinese common micro-organisms culture presevation administrative center (depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences).
(4) experimental design and statistical analysis:
1. single factor experiment:
Change successively the ratio of inoculum concentration, flour/water and vegetables/noodle soup, take DPPH clearance rate, viable count, total acid as testing index, then calculate according to the following formula its overall target:
Wherein: viable count is measured and adopted viable count in dilution-plate method working sample, and dilution factor is 10
-6, 10
-8.Each dilution factor does 3 repetitions; Above-mentioned flat board is cultivated to 24h at 30 ℃ of constant incubators, number total plate count;
Acidity assaying:
Take 25g test solution, be placed in 250ml triangular flask, add the phenolphthalein indicator of 40ml water and 0.2ml1%.Be titrated to blush 30s with 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide and do not fade, record consumes the numerical value of the volume of 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide, is denoted as V
1, same sample is measured twice; Blank test: water replaces test solution, by above-mentioned steps operation, record consumes the numerical value of the volume of 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide, is denoted as V
2; Then be calculated as follows total acid:
In formula:
X---in food the mass fraction of total acid in X, numerical value with grams per kilogram, g/kg (or g/L) represents;
The concentration of C---Standard Volumetric Solutions for Sodium Hydroxide, mol/L:
V
1---when titration test solution, consume the volume of Standard Volumetric Solutions for Sodium Hydroxide, mL;
V
2---when blank test, consume the volume of Standard Volumetric Solutions for Sodium Hydroxide, mL;
The extension rate of F---test solution;
K---the conversion coefficient of acid.The conversion coefficient of various acid is respectively: malic acid 0.067; Acetic acid 0.060; Tartaric acid 0.075; Citric acid 0.064; Containing the citric acid 0.070 () of a part crystallization water; Lactic acid 0.090; Hydrochloric acid 0.036; Phosphoric acid 0.049.
The quality of m---sample, g.
The mensuration of DPPH clearance rate:
First precision takes DPPH standard items 8mg, is placed in 100 mL volumetric flasks, and with methyl alcohol dissolving constant volume, obtaining concentration is 2 × 10
-4the DPPH standard reserving solution of mol/L, at 4 ~ 10 ℃, low temperature keeps in Dark Place for subsequent use; Then, after storing solution being mixed in the ratio of 3mL:5mL with methyl alcohol, obtain working solution; Secondly, after methyl alcohol is mixed in the ratio of 1mL:3mL with working solution, utilize UV-VIS spectrophotometry to measure absorbance in 517 nm places, be designated as A
blank=0.681; Finally, after the pulp-water of fermenting-ripening is mixed in the ratio of 1mL:3mL with methyl alcohol, get 10mL pulp-water centrifugal 10 min under 8000 r/min, obtain supernatant, after this supernatant mixes in the ratio of 1mL:3mL with working solution, reacting at normal temperature without light 20 min, survey absorbance in 517 nm places and are designated as A
sample, parallel determination 2 times, calculate as follows DPPH clearance rate:
2. response surface method optimal design:
According to single factor experiment result, with these 3 factors of ratio of inoculum concentration, flour/water and vegetables/noodle soup, utilize Design Expert 8.0 softwares to carry out experimental design (setting the results are shown in Table 1) according to Box-Behnken design principle, with flour/water X
1, vegetables/noodle soup X
2with inoculum concentration X
3for independent variable, take overall target as response y(testing program and the results are shown in Table 2), set up polynary quadratic regression equation:
Y=69.08+3.30X
1+11.27?X
2+9.99?X
3+3.92?X
1?X
2+5.93?X
1?X
3-2.31?X
2?X
3-9.18?X
1 2-4.43?X
2 2-8.44?X
3 2。
As shown in Table 3, the P < 0.01 of model, shows that regression equation is remarkable; Lose and intend P=0.2356 > 0.05, show not significantly, the quadratic regression equation predicated response value well of this test is described; To the test correction coefficient of determination R of model of regression equation
adj 2=0.8706, illustrate that this model can explain the variation of approximately 87% response; Coefficient of determination R
2=0.9438, show that equation model degree is better, test error is less; Coefficient of variation CV represents the accuracy of test, its value is less, the reliability of result of the test is higher, this test CV=8.55%, in tolerance interval, illustrate that result of the test is reliable, can replace the true point of test to carry out analysis and prediction to comprehensive nutritive index in the directly putting type fermented process of pulp-water with this model regression equation.
Table 1 experimental factor and coding schedule
Table 2 experimental design and result
The variance analysis of table 3 regression model
Note: (* P < 0.05, significant difference; * P < 0.01, difference is extremely remarkable).
0027 (5) interpretation and optimization:
Utilize Design Expert 8.0 softwares according to the analysis of drawing of polynary quadratic regression equation, obtain response surface and the contour map (referring to Fig. 1 ~ 6) thereof of regression equation.
Claims (2)
1. response surface method is optimized a preparation method for pulp-water, comprises the following steps:
(1) vegetables preparation: segment after respectively celery, lotus flower dish being cleaned, and in etc. the ratio of quality put into tank;
(2) noodle soup preparation: the ratio that the water that is 90 ~ 100 ℃ by flour and temperature is 1kg:60mL, 1 kg:80 mL, 1 kg:100 mL in horizontal gradient is respectively mixed with noodle soup;
In the tank that is placed with described vegetables respectively by being 1 kg:20 mL for horizontal gradient, 1 kg:30 mL, the ratio of 1 kg:40 mL adds described noodle soup, and the inoculum concentration that is 5%, 7%, 9% by horizontal gradient respectively accesses Mixed Microbes, be the condition bottom fermentation 4 ~ 6 days of 20 ~ 40 ℃ in temperature, can obtain the pulp-water corresponding to the ratio of different vaccination amount, flour/water and vegetables/noodle soup;
(4) experimental design and statistical analysis:
1. single factor experiment:
Change successively the ratio of inoculum concentration, flour/water and vegetables/noodle soup, take DPPH clearance rate, viable count, total acid as testing index, then calculate according to the following formula its overall target:
Wherein: viable count is measured and adopted viable count in dilution-plate method working sample, and dilution factor is 10
-6, 10
-8;
Each dilution factor does 3 repetitions; Above-mentioned flat board is cultivated to 24h at 30 ℃ of constant incubators, number total plate count;
Acidity assaying:
Take 25g test solution, be placed in 250ml triangular flask, add the phenolphthalein indicator of 40ml water and 0.2ml1%;
Be titrated to blush 30s with 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide and do not fade, record consumes the numerical value of the volume of 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide, is denoted as V
1, same sample is measured twice; Blank test: water replaces test solution, by above-mentioned steps operation, record consumes the numerical value of the volume of 0.1mol/L Standard Volumetric Solutions for Sodium Hydroxide, is denoted as V
2; Then be calculated as follows total acid:
In formula:
X---in food the mass fraction of total acid in X, numerical value with grams per kilogram, g/kg (or g/L) represents;
The concentration of C---Standard Volumetric Solutions for Sodium Hydroxide, mol/L:
V
1---when titration test solution, consume the volume of Standard Volumetric Solutions for Sodium Hydroxide, mL;
V
2---when blank test, consume the volume of Standard Volumetric Solutions for Sodium Hydroxide, mL;
The extension rate of F---test solution;
K---the conversion coefficient of acid; Wherein the conversion coefficient of various acid is respectively: malic acid 0.067; Acetic acid 0.060; Tartaric acid 0.075; Citric acid 0.064; Containing the citric acid 0.070 () of a part crystallization water; Lactic acid 0.090; Hydrochloric acid 0.036; Phosphoric acid 0.049;
The quality of m---sample, g;
The mensuration of DPPH clearance rate:
First precision takes DPPH standard items 8mg, is placed in 100 mL volumetric flasks, and with methyl alcohol dissolving constant volume, obtaining concentration is 2 × 10
-4the DPPH standard reserving solution of mol/L, at 4 ~ 10 ℃, low temperature keeps in Dark Place for subsequent use; Then, after described storing solution being mixed in the ratio of 3mL:5mL with methyl alcohol, obtain working solution; Secondly, after methyl alcohol is mixed in the ratio of 1mL:3mL with described working solution, utilize UV-VIS spectrophotometry to measure absorbance in 517 nm places, be designated as A
blank=0.681; Finally, after the pulp-water of fermenting-ripening is mixed in the ratio of 1mL:3mL with methyl alcohol, get 10mL pulp-water centrifugal 10 min under 8000 r/min, obtain supernatant, after this supernatant mixes in the ratio of 1mL:3mL with described working solution, reacting at normal temperature without light 20 min, survey absorbance in 517 nm places and are designated as A
sample, parallel determination 2 times, calculate as follows DPPH clearance rate:
2. response surface method optimal design:
According to single factor experiment result, with these 3 factors of ratio of inoculum concentration, flour/water and vegetables/noodle soup, utilize Design Expert 8.0 softwares to carry out experimental design according to Box-Behnken design principle, with flour/water X
1, vegetables/noodle soup X
2with inoculum concentration X
3for independent variable, take overall target as response y, set up polynary quadratic regression equation:
Y=69.08+3.30X
1+11.27?X
2+9.99?X
3+3.92?X
1?X
2+5.93?X
1?X
3-2.31?X
2?X
3-9.18?X
1 2-4.43?X
2 2-8.44?X
3 2；
(5) interpretation and optimization:
Utilize Design Expert 8.0 softwares according to the analysis of drawing of polynary quadratic regression equation, obtain response surface and the contour map thereof of regression equation.
2. a kind of response surface method as claimed in claim 1 is optimized the preparation method of pulp-water, it is characterized in that: described Mixed Microbes refers to that Lactobacillus brevis, Lactobacillus plantarum, lactobacillus fermenti, leuconostoc pseudomesenteroides are obtained by mixing in the ratio of 1g:1g:1g:1g; Described Lactobacillus brevis, Lactobacillus plantarum, lactobacillus fermenti, leuconostoc pseudomesenteroides are all from Chinese common micro-organisms culture presevation administrative center.
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