CN115777697A - Method for preserving postharvest Sichuan radix codonopsis - Google Patents
Method for preserving postharvest Sichuan radix codonopsis Download PDFInfo
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- CN115777697A CN115777697A CN202211429225.XA CN202211429225A CN115777697A CN 115777697 A CN115777697 A CN 115777697A CN 202211429225 A CN202211429225 A CN 202211429225A CN 115777697 A CN115777697 A CN 115777697A
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- fresh
- preservative
- sichuan
- epsilon
- polylysine
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Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
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- Seasonings (AREA)
Abstract
The invention provides a method for preserving the picked Sichuan radix codonopsitis, which comprises the steps of soaking precooled fresh Sichuan radix codonopsitis in a preservative, draining, putting the soaked fresh Sichuan radix codonopsitis in a PA/PE microporous preservation bag, extruding air in the bag, sealing and storing at-12-20 ℃; the preservative is aqueous solution of nisin and/or epsilon-polylysine. The fresh-keeping method of the invention can effectively maintain the sensory quality of the fresh codonopsis pilosula from Sichuan, reduce the weight loss rate, delay the physiological deterioration processes of aging, softening, browning and the like, effectively maintain the contents and the oxidation resistance of the major active ingredients of codonopsis pilosula from Sichuan, such as lobetyolin, atractyloide III and syringin, and effectively retain the flavor substance components of the fresh codonopsis pilosula from Sichuan in the final storage period. The method is simple to operate, green and safe, has no equipment requirement and has obvious effect. Can improve the commodity value of the codonopsis pilosula and has obvious economic benefit.
Description
Technical Field
The invention belongs to the technical field of preservation of traditional Chinese medicinal materials, and particularly relates to a postharvest preservation method for codonopsis pilosula in Sichuan province.
Background
The radix Codonopsis pilosulae is the root of radix Codonopsis pilosulae (Codonopsis tandshen Oliv) which belongs to one of the important sources of radix Codonopsis medicine collected in the Chinese pharmacopoeia of 2020 edition, is used as medicine, has mild nature and sweet taste, and has the effects of tonifying middle-jiao and Qi, nourishing blood and promoting the production of body fluid. The codonopsis pilosula is widely distributed in China and is a traditional bulk tonifying traditional Chinese medicinal material in China. The codonopsis pilosula mainly contains chemical components such as saccharides, steroids, terpenes, polyacetylene, flavone, organic acid and the like, wherein micromolecular characteristic components such as lobetyolin, syringin, atractyloide III and the like are often used as index components to evaluate the authenticity of codonopsis pilosula medicinal materials or identify varieties, years and harvesting periods. The radix codonopsitis is used as a medicine-food homologous Chinese medicinal material and has the effects of treating diseases and preserving health. However, when people eat the fresh medicinal materials, the use of the fresh medicinal materials in the field of food is greatly limited due to the limitations of mouthfeel and processing form, and the fresh medicinal materials are deeply favored by people due to the characteristics of better mouthfeel, unique flavor, rich nutrition, remarkable health-care function, convenience in eating and processing and the like. However, the best harvesting period of the codonopsis pilosula is very centralized and very short, and the harvested fresh codonopsis pilosula has strong enzyme activities, high water content, high respiratory strength and quick nutrient consumption. If the collected codonopsis pilosula is not processed and dried or kept fresh in time, physiological changes such as dehydration shrinkage, softening browning, mildew and rot and the like are easy to occur, so that the effective components in the codonopsis pilosula are further rapidly degraded, and the color, smell, taste, texture and nutritional value of the fresh codonopsis pilosula are seriously affected, thereby shortening the shelf life and limiting the development of the codonopsis pilosula industry.
At present, the fresh codonopsis pilosula is mostly sold in the market by digging and selling at the present and transporting with soil, is greatly limited by time and space, and is difficult to meet the consumption demand of the market on the fresh codonopsis pilosula. The systematic research on the preservation of the picked fresh codonopsis pilosula is almost blank, and the application of the modern preservation technology in the preservation of the fresh codonopsis pilosula is not reported for a while. How to prolong the shelf life of fresh codonopsis pilosula and improve the commercial value of the fresh codonopsis pilosula becomes a problem to be solved urgently.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a method for preserving the picked codonopsis pilosula.
The invention provides a method for preserving the picked Sichuan radix codonopsitis, which comprises the steps of soaking precooled fresh Sichuan radix codonopsitis in a preservative, draining, putting the soaked fresh Sichuan radix codonopsitis in a PA/PE microporous preservation bag, extruding air in the bag, sealing and storing at-12-20 ℃; the preservative is aqueous solution of nisin and/or epsilon-polylysine.
Preferably, in the preservative, the weight percentage concentration of nisin is 0.02-0.10%; and/or
In the preservative, the weight percentage concentration of the epsilon-polylysine is 0.02 to 0.10 percent.
Preferably, in the preservative, the weight percentage concentration of nisin is 0.05-0.08%; and/or
In the preservative, the weight percentage concentration of epsilon-polylysine is 0.05-0.10%.
Preferably, the preservative is an aqueous solution of nisin and epsilon-polylysine.
Preferably, the preservative consists of the following components in percentage by weight: 0.05% of nisin, 0.05% of epsilon-polylysine and the balance of water.
Preferably, the pre-cooling is pre-cooling at 0 ± 1 ℃.
Preferably, the soaking time is 1.5-2.5min.
Preferably, the storage is carried out at-4 to 12 ℃ after sealing.
Preferably, the sample is stored at 2 ℃ after sealing.
The invention has the following beneficial effects:
the invention adopts a response surface method to optimize the method for preserving the collected Sichuan radix codonopsis pilosulae to obtain an optimal scheme, and provides a theoretical basis for production practice. The fresh-keeping method can effectively maintain the sensory quality of the fresh codonopsis pilosula in 80 days, reduce the weight loss rate, delay the physiological deterioration processes of aging, softening, browning and the like, effectively maintain the contents and the antioxidant capacity of the main active ingredients of the codonopsis pilosula, such as lobetyolin, atractyloide III and syringin, and effectively retain the flavor substance components in the fresh codonopsis pilosula in the late storage period. The method is simple to operate, green and safe, has no equipment requirement and has obvious effect. Can improve the commodity value of the codonopsis pilosula and has obvious economic benefit.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 shows the effect of nisin concentration on the fresh-keeping effect of fresh Codonopsis Lanceolata.
FIG. 2 shows the effect of epsilon-polylysine concentration on the preservation effect of fresh Codonopsis lanceolata.
FIG. 3 shows the effect of storage temperature on the fresh-keeping effect of fresh Codonopsis Lanceolata.
FIG. 4 is a graph of the temperature (A) versus nisin concentration (B) interaction response.
FIG. 5 is a line graph showing interaction contour of temperature (A) and nisin concentration (B).
FIG. 6 is a graph of the temperature (A) versus the epsilon-polylysine concentration (C) interaction response.
FIG. 7 is a line contour plot of temperature (A) versus epsilon-polylysine concentration (C).
FIG. 8 is a graph of the interaction response curve of nisin concentration (B) and epsilon-polylysine concentration (C).
FIG. 9 is a line contour plot of the interaction of nisin concentration (B) with epsilon-polylysine concentration (C).
FIG. 10 is a graph (finger print) of volatile substances Gallery Plot of Codonopsis pilosula (Codonopsis tangshen) for different storage periods.
FIG. 11 is a graph of PCA of Codonopsis tangshen for various storage periods.
Detailed Description
The following examples are intended to facilitate a better understanding of the invention, but are not intended to limit the invention thereto. The experimental procedures in the following examples are conventional unless otherwise specified. The experimental materials used in the following examples were purchased from conventional biochemicals, unless otherwise specified.
Example 1
The screening method of the optimal preservation condition of the picked Sichuan radix codonopsis comprises the following steps:
1. single factor experiment
1. Experimental Material
Fresh Sichuan radix Codonopsis with soil was harvested and sent to the university of Lanzhou college institute of medicine (1-2 days for transportation). Screening out fresh radix Codonopsis Lanceolatae with good appearance, strong root and strip, no damage, and similar shape and size, cleaning, removing non-medicinal parts, draining off surface water, and pre-cooling in a refrigerator at 0 + -1 deg.C for 24 hr.
Precooling can eliminate field heat and prevent mildew caused by accumulation before preservation treatment; also can unify the temperature of the radix codonopsis pilosulae so as to facilitate the subsequent temperature regulation.
PA/PE microporous preservation bag (thickness 0.1mm, 5cm from the two sides of the preservation bag respectively have 8 pairs of holes with the aperture of 0.1 mm) (procured in Cangzhou Tuofei plastics Limited, size 20cm x 45 cm); nisin (purchased from Zheng Zhou Bainafo bioengineering GmbH); epsilon-polylysine (purchased from Zheng, benfo bioengineering, inc.).
2. Fresh-keeping method
2.1 study of the influence of different preservatives on the fresh-keeping effect of fresh radix Codonopsis Lanceolatae
Dividing the precooled fresh Sichuan radix Codonopsis into 6 groups, and respectively soaking the Sichuan radix Codonopsis into different concentrationsThe preservative solution (solvent is redistilled deionized water) is kept for about 2min, and the mass ratio of the feed liquid is 6:5, namely, the material-liquid ratio is to ensure that the fresh szechuan tangshen root is completely immersed in the preservative aqueous solution, the soaking is about 2min to ensure that the surface of the fresh szechuan tangshen root is completely covered by the preservative solution, the draining is carried out, the fresh szechuan tangshen root is weighed and then placed into a PA/PE microporous preservation bag, the air in the bag is extruded out, the bag is stored in a constant-temperature constant-humidity box at the temperature of 25 ℃ after being sealed, the sensory quality (color, smell, hardness and taste) and the weight loss rate of the szechuan tangshen root are measured at the storage stages of 3, 6, 9 and 12d, a certain amount of sample is taken (the sample is stored in a refrigerator at the temperature of-80 ℃ after being frozen by liquid nitrogen) at the storage stage of 12d, and H is measured 2 O 2 Content and. O 2 - The production rate was determined and each treatment was repeated 3 times, wherein the preservatives were nisin and epsilon-polylysine, respectively. The concentration gradient of each preservative solution is shown in table 1 by taking fresh Sichuan radix codonopsis which is not treated, packaged by a PA/PE microporous preservative bag and stored in a constant temperature and humidity box at 25 ℃ as a reference.
TABLE 1 treatment concentration gradient of aqueous solutions of various preservatives
2.2 study of the influence of different storage temperatures on the fresh keeping effect of fresh radix Codonopsis
Dividing the precooled fresh radix Codonopsis into 6 groups at random, weighing, placing into PA/PE microporous fresh-keeping bag with thickness of 0.2mm, extruding out air in the bag, sealing, storing at-12 deg.C, -4 deg.C, 12 deg.C, 20 deg.C, 25 deg.C, measuring sensory quality (color, smell, hardness, taste) and weight loss rate of radix Codonopsis, and taking a certain amount of sample (freezing with liquid nitrogen and storing in refrigerator at-80 deg.C) at 12d 2 O 2 Content and. O 2 - The production rate was measured by repeating each treatment 3 times without any treatment, and fresh Codonopsis tangshen (Codonopsis tangshen) was packed in PA/PE microporous freshness-retaining bags and stored in a constant temperature and humidity chamber at 25 ℃ as a control.
3. Index measurement
3.1 sensory Scoring
The appearance of fresh Sichuan radix Codonopsis is evaluated according to four grades of I, II, III and IV (excellent, good, common and poor) from four aspects of color, smell, hardness and taste, wherein the grade I corresponds to 25-21, the grade II corresponds to 20-16, the grade III corresponds to 15-11 and the grade IV corresponds to less than 10. Sensory composite score = color score + odor score + hardness score + mouthfeel score.
TABLE 2 sensory Scoring Standard Table
3.2 weight loss ratio
And measuring by adopting a weighing method. Weight loss% = (weight before storage-weight after storage)/weight before storage × 100%.
3.3 H 2 O 2 Determination of content
The experimental procedures refer to the method of Caojiankang 'guidance of physiological and biochemical experiments after fruit and vegetable harvest'. To be equivalent to H 2 O 2 The amount (. Mu.mol) is plotted on the abscissa (X) and the A415 nm value on the ordinate (Y) to yield the calibration curve Y =0.0775X +0.0003 (R) 2 = 0.9991). Per gram fresh weight of H contained in the tissue 2 O 2 As the amount of H 2 O 2 The content, expressed as mu mol/g, is calculated according to the formula (1):
wherein C is H in the sample examined by the standard curve 2 O 2 Concentration (. Mu. Mol); v t The total volume (mL) of the sample extract; v 1 Volume of sample extract (mL) used for the determination; FW is the fresh weight (g) of the sample.
3.4 ·O 2 - Production rate determination
The experimental procedures refer to the method of Wanghifei fruit and vegetable storage and processing. Taking the amount of substance (umol) corresponding to superoxide anion as abscissa (X) and A530 nm as ordinate (Y) to obtainStandard curve Y =5.1232X-0.0006 (R) 2 = 0.9998). The amount of superoxide anion produced per gram fresh weight of sample per minute was expressed as the superoxide anion production rate in nmol/(min g). The calculation formula is as follows (2):
wherein n is the amount (mu mol) of superoxide anion in the solution found by the standard curve; v is the volume (mL) of the sample extract; v s Volume (mL) of the sampled extract at the time of measurement; t is the time (min) for the sample to react with hydroxylamine; and m is the fresh weight (g) of the sample.
3.5 data analysis
WPS Office 2021 is adopted for data processing, software SPSS 22.0 is adopted for Pearson significance correlation analysis, and software GraphPad Prism 8.0.1 is adopted for data processing and drawing.
4. Analysis of results
4.1 Effect of Nisin concentration on fresh Codonopsis tangshen
FIG. 1 shows the effect of nisin concentration on the preservation effect of fresh Codonopsis lanceolata.
As can be seen from figure 1, when the nisin concentration is 0.02-0.10%, the sensory quality of fresh Sichuan radix codonopsis pilosulae can be remarkably maintained; significant reduction of O 2 - The generation rate and the delay of quality deterioration. Wherein when the nisin concentration is 0.06%, the sensory score of fresh radix Codonopsis Lanceolatae in the last storage stage is highest, the weight loss rate is lowest, and H is 2 O 2 Content and. O 2 - The production rate is lowest, and the preservation effect is best.
Influence of 4.2 epsilon-polylysine concentration on fresh-keeping effect of fresh radix Codonopsis Lanceolatae
FIG. 2 shows the effect of epsilon-polylysine concentration on the preservation effect of fresh Codonopsis lanceolata.
As can be seen from figure 2, when the concentration of the epsilon-polylysine is 0.02 to 0.10 percent, the sensory quality of the fresh Sichuan radix codonopsis pilosulae can be remarkably maintained; significant reduction of O 2 - Producing a rate and delaying quality degradation. It is composed ofWhen the concentration of the middle epsilon-polylysine is 0.06 percent, the sensory score of fresh radix codonopsis pilosulae in the final storage stage is highest, the weight loss rate is lowest, and H is 2 O 2 Content and. O 2 - The production rate is lowest, and the preservation effect is best.
4.3 Effect of storage temperature on fresh Codonopsis tangshen
FIG. 3 shows the effect of storage temperature on the fresh-keeping effect of fresh Codonopsis Lanceolata.
As can be seen from figure 3, when the storage temperature is-12-20 ℃, the sensory quality of the fresh Sichuan radix codonopsis can be remarkably maintained; remarkably reducing weight loss rate and O 2 - Producing a rate and delaying quality degradation. Wherein when the storage temperature is 4 ℃, the sensory score of the fresh Sichuan radix codonopsis pilosulae in the final storage stage is highest, the weight loss rate is lowest, and H 2 O 2 Content and. O 2 - The production rate is low, and the preservation effect is best.
In conclusion, the optimal treatment concentrations and the optimal storage temperatures of the two preservatives are selected through the single-factor experiment. When the concentration of nisin and epsilon-polylysine is 0.02-0.10% and the storage temperature is-12-20 ℃, the storage time of the fresh Sichuan radix codonopsis can be remarkably prolonged, and the first-grade fresh Sichuan radix codonopsis is still the second-grade better Sichuan radix codonopsis in the final storage stage.
2. Response surface experiment
1. Experimental Material
As in the single factor experiment.
2. Fresh-keeping method
On the basis of 3 single-factor experiments, design-Expert 11 software is used for designing a response surface scheme. The storage temperature (A), the nisin concentration (B) and the epsilon-polylysine concentration (C) are respectively 3 independent variables, and the weight coefficient of each index is determined by adopting an analytic hierarchy process-index correlation-based weight determination method (AHP-CRITIC) to obtain sensory score, weight loss rate, effective component indexes (lobetyolin, syringin and atractyloide III), antioxidant activity indexes (total flavonoids, total phenols and DPPH free radical clearance rate) and aging-related physiological indexes (H) 2 O 2 Content,. O 2 - Production rate, malondialdehyde content, superoxide dismutase, hydrogen peroxideEnzyme), browning related physiological indexes (polyphenol oxidase and peroxidase), and softening related physiological indexes (pectin methylesterase, polygalacturonase, cellulase and beta-glucosidase), and taking the comprehensive score as a response value. The optimal value in each single-factor experiment was selected to be a zero level, and response surface experiments were performed for 17 combinations of treatments. The center and level of the experimental design are shown in table 3, and the specific grouping condition and corresponding comprehensive score of each treatment combination are shown in table 4.
TABLE 3 independent variables and their code values in response surface analysis experiments
TABLE 4 response surface Experimental groups and composite scores
Randomly dividing the precooled fresh Sichuan radix codonopsis into 6 groups, and immersing the groups into the mixed preservative for 2min, wherein the material-liquid ratio is 6: and 5, draining, weighing, putting into a PA/PE microporous fresh-keeping bag, extruding air out of the bag, sealing, storing at a corresponding temperature, sampling at 80d in the storage period, measuring the 19 indexes, and repeating the treatment for 3 times.
The preparation method of the mixed preservative comprises the following steps: and calculating and weighing nisin and epsilon-polylysine according to the mass ratio of the feed liquid, dissolving nisin and epsilon-polylysine in water, and uniformly stirring for later use.
3. Index measurement
3.1 sensory Scoring
As in the single factor experiment.
3.2 weight loss ratio
The same as single factor experiment.
3.3 H 2 O 2 Content (wt.)
As in the single factor experiment.
3.4 ·O 2 - Rate of generation
The same as single factor experiment.
3.5 cell wall degrading enzyme Activity assay
The Pectin Methylesterase (PME), polygalacturonase (PG), cellulase (Cx) and beta-glucosidase (beta-glu) activity are measured by referring to the Caojiakang physiological and biochemical experiment guidance after fruit and vegetable harvest. Taking the amount of glucose as abscissa (X) and the A540 nm value as ordinate (Y), a calibration curve Y =0.6335X-0.0068 (R) 2 ＝0.9992)。
PG enzyme activity units are defined as the amount of enzyme required to release 1mg of galacturonic acid per 1g fresh weight per 1h as one enzyme activity unit (U/g FW). PME enzyme activity units are defined as the amount of enzyme required to release 1mg galacturonic acid per 1g fresh weight per 1h as one enzyme activity unit (U/g FW). Cx enzyme Activity units are defined as the amount of enzyme required to release 1mg reducing sugars per 1g fresh weight per 1h as one enzyme activity unit (U/g FW). The β -glu enzyme activity unit is defined as the amount of enzyme required to release 1mg of reducing sugar per 1g fresh weight per 1h as one enzyme activity unit (U/g FW).
3.6 measurement of other physiological indices
Malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, catalase (CAT) activity, polyphenol oxidase (PPO) activity and Peroxidase (POD) activity were determined with reference to the test kit instructions.
3.7 determination of the content of lobetyolin, syringin and atractyloide III
3.7.1 preparation of test solutions
Taking 4g of fresh Sichuan radix codonopsitis powder, precisely weighing, placing in a conical flask with a plug, adding 50mL of methanol, carrying out ultrasonic extraction for 45min, cooling, filtering, concentrating in water bath to dryness, then re-dissolving to 2mL with methanol, shaking up, and filtering with a 0.45-micron filter membrane to obtain a stock solution of a sample to be detected. And (4) injecting samples according to chromatographic conditions to respectively determine atractyloide III and syringin. Precisely absorbing 200 mu L of stock solution of a sample to be measured, adding methanol to dilute the stock solution to a constant volume of 1mL, shaking up, filtering with a 0.45 mu m filter membrane to obtain the sample to be measured, and carrying out sample injection according to chromatographic conditions to measure the lobetyolin.
3.7.2 drawing of Standard Curve
Radix Codonopsis alkynil standard curve: radix Codonopsis alkyne glycoside is used as reference substance to prepare radix Codonopsis alkyne glycoside stock solution with concentration of 1.042 mg/ml. It was then diluted with methanol to 32.56, 65.13, 130.25, 260.50, 521.00, 1042.00 μ g/mL series of graded lobetyolin control solutions. Chromatographic conditions are as follows: volume ratio acetonitrile: water =26:74 at a flow rate of 1.0 mL/min -1 The detection wavelength is 267nm, the column temperature is 30 ℃, the sample is added by 10 mu L, and a regression equation is drawn by taking the concentration of a reference substance as a horizontal coordinate (X) and the peak area as a vertical coordinate (Y) and is Y =8.5547X +65.207 (R) 2 ＝0.9993)。
Syringin standard curve: syringin is used as a reference substance to prepare a syringin stock solution with the concentration of 1.044 mg/ml. It was then diluted with methanol to 6.53, 13.05, 26.10, 52.20, 104.40 μ g/mL series of graded syringin control solutions. The chromatographic conditions are as follows: the mobile phase is acetonitrile (A) -0.2% by volume of acetic acid water solution (B), and the gradient elution is carried out by the following steps: 0-10 min, 10% (A) by volume; 10-20 min, 15% (A) volume percentage; 20-30 min, 10 percent of volume (A) and 1.0 mL/min of flow rate -1 Detecting wavelength of 220nm, column temperature of 30 ℃, sample feeding amount of 20 mu L, and drawing a regression equation of Y =90.613X +75.282 (R) by taking the concentration of a reference substance as a horizontal coordinate (X) and the peak area as a vertical coordinate (Y) 2 ＝1)。
Atractylenolide III standard curve: atractylodes lactone III is used as a reference substance to prepare atractylis lactone III stock solution with the concentration of 1.238 mg/ml. It was then diluted with methanol to 0.31, 1.24, 6.19, 30.95, 309.50, 619.00, 1238.00 μ g/mL series of graded atractylenolide iii control solutions. The chromatographic conditions are volume ratio acetonitrile: water =71:29 at a flow rate of 1.0 mL/min -1 Detecting wavelength of 220nm, column temperature of 30 deg.C, sample feeding amount of 20 μ L, and drawing regression equation with reference substance concentration as abscissa (X) and peak area as ordinate (Y) as Y =43.536X +18.169 (R) 2 ＝0.9999)。
3.8 measurement of Total Flavonoids, total phenols content and DPPH radical scavenging ability
3.8.1 preparation of test solutions
Precisely weighing 2g of fresh Sichuan radix Codonopsis lyophilized powder, placing in a 150mL conical flask with a plug, performing ultrasonic extraction for 2 times and 30min each time by using 25mL 70% ethanol as a solvent, performing suction filtration on the filtrate to a 100mL volumetric flask, fixing the volume to a scale, shaking up, and filtering with a microporous membrane to obtain a subsequent filtrate, namely a sample solution.
3.8.2 drawing of Standard Curve
Standard curve of total flavonoids: accurately weighing 2mg of rutin standard substance dried at 105 deg.C to constant weight, dissolving with appropriate amount of anhydrous ethanol, metering volume to 10mL, shaking to obtain rutin standard substance stock solution with concentration of 0.2000g/L, and storing at 4 deg.C for use. Precisely measuring rutin control solution 0.4,0.8,1.2,1.6,2.0 and 2.4mL, respectively placing in 10mL measuring bottles, adding water to 2.4mL, adding 5% sodium nitrite solution 0.4mL, shaking, standing for 6min, adding 10% aluminum nitrate 0.4mL, shaking, standing for 6min, adding 4% sodium hydroxide solution 4mL, adding water to scale, shaking, standing for 15min, and making into control solution with rutin concentrations of 0.008,0.016,0.024,0.032,0.040 and 0.048g/L in sequence. Taking A505 nm as ordinate (Y), concentration as abscissa (X), and standard curve of Y =9.6571X-0.0164 (R) 2 ＝0.9991)。
Total phenol standard curve: accurately weighing 1mg of gallic acid standard, dissolving with water, diluting to 10mL, shaking, diluting 1mL with distilled water 8 times, and making into 0.0125g/L reference standard solution. Accurately sucking 0.8,1.6,2.4,3.2,4.0 and 4.8mL of standard solution, respectively placing the standard solution into a 10mL measuring flask, adding distilled water to 2.0mL, uniformly mixing, adding 1mL of Freund's reagent, after 3min, adding 10% Na with weight percentage 2 CO 3 Diluting the solution 2mL with water to 10mL, heating in 30 deg.C water bath for 30min, taking out, shaking to obtain control solution with gallic acid concentration of 0.0010,0.0020,0.0030,0.004,0.005, and 0.006g/L. Taking A765 nm as ordinate (Y) and concentration as abscissa (X), the calibration curve is Y =217.31X +0.0761, (R) 2 ＝0.9991)。
3.8.3 sample assays
And (3) total flavone content determination: precisely measuring 2mL of the test solution, measuring absorbance according to the operation under the item of a standard curve, and calculating the content of the total flavonoids in the sample according to the standard curve.
And (3) total phenol content determination: precisely measuring 1.5mL of the test solution, operating according to the standard curve, measuring absorbance, and calculating the total phenol content in the sample according to the standard curve.
DPPH free radical clearance determination: taking DPPH 4mg, placing in a 100mL measuring flask, adding absolute ethyl alcohol to a constant volume to obtain 0.1mmol/L stock solution, and placing at 4 ℃ for later use. Transferring 100 μ L of the sample solution and 2mL of the stock solution, placing in the same test tube, shaking, reacting in a dark room for 30min, and measuring the absorbance value As at wavelength of 517 nm. 2mL of the stock solution and 100. Mu.L of absolute ethanol were transferred and placed in the same test tube, shaken, and the absorbance Ab of the mixed solution was measured. Transferring 2mL of absolute ethyl alcohol and 100 mu L of test solution, placing the absolute ethyl alcohol and the test solution in the same test tube, measuring the absorbance Ac of the mixed solution, and calculating the clearance rate, wherein the formula is as shown in (3):
3.9 data analysis
The same as single factor experiment.
4. Analysis of results
4.1 Determining each index weight by AHP-CRITIC mixed weighting method
According to the Box-Behnken experiment Design principle, design Expert 11 software is used for carrying out three-factor three-level response surface experiment Design by taking temperature, nisin concentration and epsilon-polylysine concentration as experiment factors to obtain 17 treatment combinations, after fresh Sichuan radix codonopsis pilosulae is subjected to corresponding fresh-keeping treatment and stored, the 80 th day of the storage period is sampled, and the 19 indexes are measured. The measurement results are shown in tables 5 to 7.
TABLE 5 influence of different fresh-keeping treatments on sensory score, weight loss rate, effective components and antioxidant activity of fresh radix Codonopsis
TABLE 6 influence of different fresh-keeping treatments on the physiological indexes of oxidation aging and browning of fresh radix Codonopsis
TABLE 7 influence of different fresh-keeping treatments on the physiological index of fresh radix Codonopsis Lanceolatae
Determining the 19 index weights by adopting an AHP-CRITIC mixed weighting method, wherein the comprehensive weight calculation formula of the mixed weighting method is as follows (4):
the data were processed according to the SPSS 22.0 software, and the 19-term index integrated weight was calculated according to the formula shown in Table 8.
TABLE 8 weight analysis results
And carrying out comprehensive grading comparison on the processed experimental data by adopting weight coefficients obtained by calculation of an AHP, CRITIC and AHP-CRITIC mixed weighting method. The correlation coefficients of the CRITIC and AHP, the CRITIC and AHP-CRITIC mixed weighting method and the AHP and AHP-CRITIC mixed weighting method are respectively 0.926,0.928 and 0.998, the correlation of the three methods is extremely obvious (P < 0.01), and the scoring results of the 3 methods are consistent.
From the weight coefficient analysis, the correlation coefficient of CRITIC and AHP is-0.038, and the correlation is not significant (P =0.878> < 0.05), which indicates that the information reflected by the CRITIC and the AHP does not have the superposition.
The AHP-CRITIC mixed weighting method is considered from the main aspect and the objective aspect, the embodied information is more comprehensive, and the method is more scientific and reasonable than a single method.
4.2 regression model construction and analysis of variance
After 17 sets of processed composite scores were obtained (see Table 4 above), regression fitting was performed using Design-Expert 11 software. And obtaining a quadratic polynomial fitting equation of independent variable storage temperature (A), nisin concentration (B), epsilon-polylysine concentration (C) and dependent variable comprehensive score (Y).
Y＝60.47-9.79*A-2.99*B-3.75*C+2.49*AB-0.2668*AC+3.56*BC-20.50*A 2 -9.14*B 2 -11.56*C 2 。
The results of the anova are shown in Table 9.
TABLE 9 analysis of variance of response surface
Note: p is<0.01 is extremely significant, P<0.05 is significant: r is 2 ＝0.9572，R 2 Adj ＝0.9022，CV＝12.26％。
From the data results in table 9, it can be seen that the model regression equation is significant, the misfit term is not significant, and the fitting degree of the regression equation is good, which has statistical significance. Determining the coefficient R 2 A value of 0.9572 indicates that the measured value is closer to the predicted value. Correction factor R 2 Adj Is 0.9022, which shows that the three factors have 90.22 percent of influence on the fresh-keeping effect of the fresh codonopsis pilosula, and shows that the experiment has higher accuracy. The coefficient of variation CV was 12.26%, which was less than 15%, indicating good stability of the experiment.
According to the value of P, the quality guarantee period of fresh Sichuan radix codonopsis is influenced by the following factors: a > C > B, and the quadratic term A 2 、B 2 And C 2 The effect on composite score was very significant.
4.3 two-factor interaction response surface analysis
The response surface graph and the contour graph of the influence of the 3 interactive items are shown in fig. 4-9, which shows that the level selection of each experimental factor is reasonable, wherein the influence of the interaction of the temperature and the two preservatives on the comprehensive score at the final storage stage of the fresh codonopsis pilosula is stronger, and the influence of the interaction between the two preservatives on the comprehensive score is second, and is consistent with the result of the P value.
4.4 validation of optimal conditions
The optimal method for obtaining the storage and preservation of the collected codonopsis pilosula in the szechuan tangshen by using Design-Expert 11 software comprises the following steps: the mixed preservative contains 0.05 percent of nisin and 0.05 percent of epsilon-polylysine, and the storage temperature is 2 ℃. Three verification experiments are carried out under the condition, the 19 indexes are measured on the 80 th day of storage, the results are shown in table 10, and the comprehensive scores are calculated according to the AHP-CRITIC mixed weight in table 8 and are respectively 66.8810, 63.5256 and 60.7902, the average value is 63.7323 and is closer to the model predicted value of 62.4208, so that the degree of fitting of the equation is good, the reliability of the obtained optimal fresh-keeping method is high, and the optimal fresh-keeping method can be used for industrial fresh-keeping storage of fresh radix codonopsis pilosulae.
Table 10 verifies the results of the experiment
Example 2
A method for storing and preserving the collected Sichuan radix codonopsis comprises the following steps:
(1) Immersing the precooled fresh Sichuan radix codonopsis in a mixed preservative for 2min, wherein the material-liquid ratio is 6:5, wherein the concentration of nisin in the mixed preservative is 0.05%, and the concentration of epsilon-polylysine is 0.05%;
(2) Draining radix Codonopsis, weighing, placing into PA/PE microporous fresh-keeping bag, squeezing out air, sealing, and storing at 2 deg.C.
Example 3
And (3) under the optimal preservation condition (namely, the preservation is carried out by adopting the mode of example 2), measuring the volatile compounds in the final storage stage of the fresh codonopsis pilosula.
1. Material
Fresh szechuan tangshen root sample (S0); storing a 40 th Sichuan radix codonopsis sample (LPN 40) under the optimal fresh-keeping condition; storing an 80 th Sichuan radix Codonopsis sample (LPN 80) under optimal fresh-keeping conditions; storing a 3 rd Chuanshen sample (RT 3) at room temperature and storing a 6 th Chuanshen sample (RT 6) at room temperature.
2. Method of producing a composite material
2.1 instruments
Flavour
Flavor analyzer (GAS phase-ion mobility spectrometry), GAS, germany.
2.2 sample treatment
1g of the sample was placed in a 20mL headspace bottle, incubated at 60 ℃ for 15 minutes, and then injected.
2.3 GC-IMS conditions
Automatic headspace sampler: the sample volume is 500 mu L, the incubation time is 15min, the incubation temperature is 60 ℃, and the analysis time is 20min. A chromatographic column: MXT-WAX (30 m. Times.0.53 mm. Times.1 μm), column temperature: 60 ℃; carrier gas/drift gas: high purity nitrogen (99.999%); drift velocity: 150mL/min; carrier gas flow rate: 2mL/min (0-2 min), 2-10 mL/min (2-10 min), 10-100 mL/min (10 min-20 min); temperature of ion transfer tube: 45 ℃; temperature of the sample injection needle: 85 ℃.
3. Analysis of results
The fingerprint of the volatile substances of 5 stored Codonopsis tangshen is shown in FIG. 10, and 102 volatile organic compounds, including 9 esters, 23 aldehydes, 16 alcohols, 9 ketones, 1 acid, 1 thiophene, 1 furan and 42 unknown compounds, were detected.
The aldehydes and alcohols are the largest in the quantitative volatiles. The main volatile of fresh radix Codonopsis Lanceolatae is n-hexanal, (E) -2-hexenal, etc., wherein n-hexanal is the main component of its special fragrance. From the results, it can be seen that the content of n-hexanal is gradually reduced after storage and preservation, and the preservation method described in example 2 can maintain good quality of radix codonopsis aroma to some extent.
FIG. 10 is a graph (finger print) of volatile substances Gallery Plot of Codonopsis pilosula (Codonopsis tangshen) stored for different periods.
Consistent with the conclusions of the Gallery Plot, it can be seen from fig. 11 that 5 samples can be distinguished by PCA analysis. Wherein LPN40 and LPN80 are closer to S0, and the overall volatile components are similar. Further shows that the fresh keeping method in the embodiment 2 can better keep the volatile flavor substances of the fresh szechuan tangshen root in the storage process.
FIG. 11 is a graph of PCA of the volatile substances from Codonopsis tangshen during different storage periods.
Example 4
Compared with the embodiment 2, the mixed preservative has the nisin concentration of 0.06 percent and the epsilon-polylysine concentration of 0.06 percent, and is stored at 4 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 5
Compared with the embodiment 2, the mixed preservative has the nisin concentration of 0.02 percent and the epsilon-polylysine concentration of 0.06 percent, and is stored at 4 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 6
Compared with the embodiment 2, the mixed preservative has the nisin concentration of 0.02 percent and the epsilon-polylysine concentration of 0.05 percent, and is stored at the temperature of 2 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 7
Compared with the embodiment 2, the mixed preservative has the nisin concentration of 0.08 percent and the epsilon-polylysine concentration of 0.06 percent, and the PA/PE microporous preservative bag is packaged and then stored at 0 ℃.
Example 8
Compared with the embodiment 2, the adopted storage and preservation method of the codonopsis pilosula in the szechuan tangshen root has the advantages that the concentration of nisin in the preservative is 0.08 percent, and the codonopsis pilosula in the preservative is stored at 2 ℃ after being packaged by PA/PE microporous preservative bags.
Example 9
Compared with the embodiment 2, the adopted storage and preservation method of the codonopsis pilosula in which the nisin concentration is 0.04 percent is adopted, and the codonopsis pilosula in which the nisin is packaged by PA/PE microporous preservation bags is stored at the temperature of-4 ℃.
Example 10
Compared with the embodiment 2, the method for storing and preserving the collected Sichuan radix codonopsis pilosulae has the advantages that the concentration of epsilon-polylysine in the preservative is 0.06 percent, and the Sichuan radix codonopsis pilosulae is stored at 0 ℃ after being packaged by a PA/PE micropore preservative bag.
Example 11
Compared with the embodiment 2, the method for storing and preserving the collected Sichuan radix codonopsis pilosulae has the advantages that the concentration of epsilon-polylysine in the preservative is 0.04 percent, and the Sichuan radix codonopsis pilosulae is stored at 12 ℃ after being packaged by a PA/PE microporous preservative bag.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. 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 (9)
1. A method for preserving the picked Sichuan radix codonopsis pilosulae is characterized by comprising the following steps: soaking the precooled fresh Sichuan radix codonopsis in a preservative, draining, putting into a PA/PE microporous preservative bag, extruding air in the bag, sealing and storing at-12-20 ℃; the preservative is aqueous solution of nisin and/or epsilon-polylysine.
2. The method of claim 1, wherein: in the preservative, the weight percentage concentration of nisin is 0.02-0.10%; and/or
In the preservative, the weight percentage concentration of the epsilon-polylysine is 0.02 to 0.10 percent.
3. The method of claim 1, wherein: in the preservative, the weight percentage concentration of nisin is 0.05-0.08%; and/or
In the preservative, the weight percentage concentration of epsilon-polylysine is 0.05-0.10%.
4. The method of claim 3, wherein: the preservative is aqueous solution of nisin and epsilon-polylysine.
5. The method of claim 4, wherein: the preservative comprises the following components in percentage by weight: 0.05% of nisin, 0.05% of epsilon-polylysine and the balance of water.
6. The method of claim 1, wherein: the pre-cooling is performed at 0 +/-1 ℃.
7. The method of claim 1, wherein: the soaking time is 1.5-2.5min.
8. The method of claim 1, wherein: sealing and storing at-4-12 deg.C.
9. The method of claim 8, wherein: sealed and stored at 2 ℃.
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