CN115777697B - Method for preserving radix codonopsis pilosulae after picking - Google Patents
Method for preserving radix codonopsis pilosulae after picking Download PDFInfo
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- CN115777697B CN115777697B CN202211429225.XA CN202211429225A CN115777697B CN 115777697 B CN115777697 B CN 115777697B CN 202211429225 A CN202211429225 A CN 202211429225A CN 115777697 B CN115777697 B CN 115777697B
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Classifications
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- 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 radix codonopsis pilosulae after picking, which comprises the steps of soaking pre-cooled fresh radix codonopsis pilosulae in preservative, draining, putting into 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. The fresh-keeping method can effectively maintain the sensory quality of the fresh radix codonopsis pilosulae within 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 antioxidation capability of the main active components of the radix codonopsis pilosulae, namely the radix codonopsis pilosulae acetylenic glycoside, the atractylenolide III and the syringin, and effectively retain the flavor components in the fresh radix codonopsis pilosulae at the end of storage. The method is simple to operate, green and safe, has no equipment requirement and has obvious effect. Can improve the commodity value of the dangshen and has remarkable economic benefit.
Description
Technical Field
The invention belongs to the technical field of preservation of traditional Chinese medicinal materials, and particularly relates to a method for preserving radix codonopsis pilosulae after picking.
Background
The radix codonopsis pilosulae is root of radix codonopsis pilosulae (Codonopsis tangshen Oliv) belonging to the root of radix codonopsis pilosulae belonging to the family of Campanulaceae, belonging to one of important sources of the radix codonopsis pilosulae in the Chinese pharmacopoeia of 2020 edition, and has the effects of tonifying middle-jiao, replenishing qi, nourishing blood and promoting salivation by taking the root as a medicine. The codonopsis pilosula is widely distributed in China and is a traditional bulk tonic Chinese medicinal material in China. The radix Codonopsis mainly contains saccharide, steroid, terpenoid, polyacetylene, flavone, organic acid, etc., wherein small molecular characteristic components such as radix Codonopsis acetylenic glycoside, syringin and atractylenolide III are commonly used as index components for evaluating the authenticity of radix Codonopsis medicinal material or identifying variety, age and harvest time. The radix codonopsis is used as a traditional Chinese medicine with the same medicine and food functions and has the effects of treating diseases and preserving health. However, when eating, the dry medicinal materials are greatly limited in use in the food field due to the limitations of taste and processing form, and the fresh medicinal materials are favored by people due to the characteristics of better taste, unique flavor, rich nutrition, obvious health care function, convenient eating and processing and the like. However, the best harvesting period of the dangshen is very concentrated and short, the activity of various enzymes of the fresh dangshen after harvesting is strong, the water content is high, the respiration intensity is high, and the nutrient component consumption is fast. If the fresh dangshen is not timely processed and dried or preserved after being picked, physiological changes such as water loss shrinkage, softening and browning, mildew and rot are easy to occur, further effective components in the dangshen are rapidly degraded, the color, smell, taste, texture and nutritive value of the fresh dangshen are seriously affected, and therefore the shelf life is shortened, and the development of the dangshen industry is limited.
At present, fresh dangshen is usually excavated and sold at present in the market, and is sold in a mode of carrying out soil transportation, so that the limitation of time and space is very large, and the consumption requirement of the market on the fresh dangshen is difficult to meet. The systematic research on fresh-keeping of fresh dangshen after picking is almost blank, and the application of the modern fresh-keeping technology in fresh-keeping of fresh dangshen is not reported temporarily. How to prolong the shelf life of fresh dangshen and improve the commercial value of the dangshen is 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 radix codonopsis pilosulae after picking.
The invention provides a method for preserving radix codonopsis pilosulae after picking, which comprises the steps of soaking pre-cooled fresh radix codonopsis pilosulae in preservative, draining, putting into 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.
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 epsilon-polylysine is 0.02-0.10%.
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: nisin 0.05%, epsilon-polylysine 0.05% and water balance.
Preferably, the pre-cooling is performed at 0+ -1deg.C.
Preferably, the soaking time is 1.5-2.5min.
Preferably, the sealing is followed by storage at-4-12deg.C.
Preferably, the container is stored at 2℃after sealing.
The beneficial effects of the invention are as follows:
the invention optimizes the method for preserving the radix codonopsis pilosulae after picking by adopting a response surface method 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 radix codonopsis pilosulae within 80 days, reduce the weight loss rate, delay the physiological deterioration processes of aging, softening, browning and the like, effectively maintain the content and the antioxidation capability of the main active components of the radix codonopsis pilosulae, namely the radix codonopsis pilosulae acetylenic glycoside, the atractylenolide III and the syringin, and effectively retain the flavor components in the fresh radix codonopsis pilosulae at the end of storage. The method is simple to operate, green and safe, has no equipment requirement and has obvious effect. Can improve the commodity value of the dangshen and has remarkable economic benefit.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate the invention and together with the embodiments of the invention, serve to explain the invention. In the drawings:
FIG. 1 shows the effect of nisin concentration on fresh Sichuan radix Codonopsis.
FIG. 2 shows the effect of epsilon-polylysine concentration on fresh radix Codonopsis Lanceolatae preservation effect.
FIG. 3 shows the effect of storage temperature on the fresh keeping effect of fresh Sichuan radix Codonopsis.
FIG. 4 is a graph showing the response of temperature (A) to nisin concentration (B).
FIG. 5 is a plot of temperature (A) versus nisin concentration (B).
FIG. 6 is a graph showing the response of the interaction of temperature (A) with epsilon-polylysine concentration (C).
FIG. 7 is a contour plot of temperature (A) versus epsilon-polylysine concentration (C).
FIG. 8 is a graph showing the response of nisin concentration (B) to epsilon-polylysine concentration (C).
FIG. 9 is a plot of the interaction contour of nisin concentration (B) with epsilon-polylysine concentration (C).
Fig. 10 shows the Plot (fingerprint) of volatile substances gallry Plot of radix codonopsis pilosulae at different storage periods.
FIG. 11 PCA graphs of volatile materials of Sichuan radix Codonopsis during different storage periods.
Detailed Description
The following examples facilitate a better understanding of the present invention, but are not intended to limit the same. The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the examples described below, unless otherwise specified, were purchased from conventional biochemistry reagents.
Example 1
The screening method of the optimal fresh-keeping condition of the radix codonopsis pilosulae after picking comprises the following steps:
1. single factor experiment
1. Experimental materials
The fresh szechwon tangshen root is picked up and sent to the university of Lanzhou college of medicine (1-2 days of transportation) with soil. Screening fresh radix codonopsis pilosulae with good appearance, strong root, no worm damage and mildew, no damage and similar shape and size, cleaning, removing non-medicinal parts, draining surface water, and pre-cooling the medicinal materials at 0+ -1deg.C for 24 hr.
The pre-cooling can eliminate field heat and prevent mildew caused by accumulation before preservation treatment; the temperature of the szechuan pilose asiabell root can be unified, so that the subsequent temperature regulation is convenient.
PA/PE microporous fresh-keeping bag (thickness 0.1mm, 5cm apart from two sides of fresh-keeping bag respectively have 8 pairs of holes with aperture 0.1 mm) (purchased from Cangzhou Tuofeiling Co., ltd., size 20cm×45 cm); nisin (purchased from bioengineering, inc. Of bayer buddha, zheng); epsilon-polylysine (purchased from biological engineering Co., ltd., bayer, zhengzhou).
2. Fresh-keeping method
2.1 study of the Effect of different antistaling Agents on fresh Sichuan radix Codonopsis
Dividing the pre-cooled fresh radix codonopsis pilosulae into 6 groups at random, and respectively immersing the radix codonopsis pilosulae in preservative aqueous solutions (the solvent is distilled deionized water) with different concentrations for about 2min, wherein the mass ratio of the feed liquid is 6:5, namely, the feed-liquid ratio is to ensure that the fresh radix codonopsis pilosulae is fully immersed in the aqueous solution of the preservative, the soaking is carried out for about 2 minutes, the surface of the fresh radix codonopsis pilosulae is fully covered by the preservative, the fresh radix codonopsis pilosulae is drained, the fresh radix codonopsis pilosulae is weighed and put into a PA/PE microporous preservative bag, air in the bag is extruded, the bag is sealed and stored in a constant temperature and humidity box at 25 ℃, the sensory quality (color, smell, hardness and taste) and the weight loss rate of the radix codonopsis pilosulae are measured at 3 rd, 6 th, 9 th and 12d of storage, and a certain amount of samples (stored in a refrigerator at-80 ℃ after liquid nitrogen freezing) are taken at 12d of storage, and the mixture is subjected to H 2 O 2 Content sum O 2 - The production rate was measured and each treatment was repeated 3 times, wherein the preservative was nisin and epsilon-polylysine, respectively. The PA/PE microporous fresh-keeping bag is packaged and stored at 25 ℃ without any treatmentThe concentration gradient of the fresh radix codonopsis pilosulae in the temperature and humidity box is shown in table 1.
TABLE 1 treatment concentration gradient of aqueous solutions of respective antistaling agents
2.2 study of the influence of different storage temperatures on the fresh-keeping Effect of fresh Sichuan radix Codonopsis
Dividing pre-cooled fresh radix Codonopsis Lanceolatae into 6 groups, weighing, placing into 0.2mm thick PA/PE microporous fresh-keeping bag, extruding air in the bag, sealing, storing at-12deg.C, -4deg.C, 12deg.C, 20deg.C, 25deg.C, measuring sensory quality (color, smell, hardness, taste) and weight loss rate of radix Codonopsis Lanceolatae at 3, 6, 9, and 12d, collecting a certain amount of sample (stored in-80deg.C refrigerator after freezing with liquid nitrogen) at 12d, and collecting H 2 O 2 Content sum O 2 - The production rate was measured and each treatment was repeated 3 times, with fresh radix codonopsis pilosulae packaged in PA/PE microporous fresh-keeping bags and stored in a constant temperature and humidity cabinet at 25 ℃ as a control.
3. Index measurement
3.1 sensory scoring
The appearance of fresh Sichuan pilose asiabell root is evaluated according to four grades of I, II, III and IV (excellent, good, general and poor), wherein grade I corresponds to 25-21, grade II corresponds to 20-16, grade III corresponds to 15-11 and grade IV corresponds to less than 10. Sensory integrated score = color score + odor score + hardness score + mouthfeel score.
TABLE 2 sensory scoring criteria
3.2 weight loss ratio
The measurement is carried out by a weighing method. Weight loss% = (weight before storage-weight after storage)/weight before storage×100%.
3.3 H 2 O 2 Content determination
Experimental procedures refer to Cao Jiankang "physiological and Biochemical Experimental guidance after fruit and vegetable harvest". To be equivalent to H 2 O 2 The quantity (. Mu. Mol) is on the abscissa (X), the A415 nm value is on the ordinate (Y), the standard curve Y=0.0775X+0.0003 (R) 2 = 0.9991). In terms of H contained in per gram of fresh weight tissue 2 O 2 Is taken as H 2 O 2 Content expressed as μmol/g, calculated as (1):
wherein C is H in a sample of standard curve examination 2 O 2 Concentration (. Mu. Mol); v (V) t Total volume of sample extract (mL); v (V) 1 Sample extract volume (mL) for measurement; FW is the fresh weight of the sample (g).
3.4 ·O 2 - Production rate determination
Experimental procedures reference Wang Hongfei methods of fruit and vegetable storage and processing. The amount (umol) of the substance corresponding to the superoxide anion is taken as an abscissa (X), the A530 nm value is taken as an ordinate (Y), and a standard curve Y= 5.1232X-0.0006 (R 2 =0.9998). The amount of the superoxide anion generating substance per minute per gram of fresh weight sample was expressed as nmol/(min.g) as the superoxide anion generating rate. The calculation formula is as follows (2):
wherein n is the amount (mu mol) of the superoxide anion substance in the solution as determined by the standard curve; v is the volume (mL) of the sample extract; v (V) s To determine the volume (mL) of the sample extract; t is the time (min) for the sample to react with hydroxylamine; m is the fresh weight (g) of the sample.
3.5 data analysis
Data processing is performed by using WPS Office 2021, pearson significance correlation analysis is performed by using SPSS 22.0 software, and data processing and drawing are performed by using GraphPad Prism 8.0.1 software.
4. Analysis of results
4.1 Effect of nisin concentration on fresh Changshen fresh-keeping Effect
FIG. 1 shows the effect of nisin concentration on fresh Sichuan radix Codonopsis.
As can be seen from fig. 1, the sensory quality of fresh codonopsis pilosula can be remarkably maintained when the concentration of nisin is 0.02-0.10%; significantly reduce O 2 - The rate of production delays deterioration of quality. Wherein, when the concentration of nisin is 0.06%, the fresh radix codonopsis pilosulae has the highest sensory score at the end of storage and the lowest weight loss rate and H 2 O 2 Content sum O 2 - The production rate is the lowest and the fresh-keeping effect is the best.
Influence of 4.2 epsilon-polylysine concentration on fresh radix codonopsis preservative effect
FIG. 2 shows the effect of epsilon-polylysine concentration on fresh radix Codonopsis Lanceolatae preservation effect.
As can be seen from fig. 2, the sensory quality of fresh codonopsis pilosula can be remarkably maintained when the concentration of epsilon-polylysine is 0.02-0.10%; significantly reduce O 2 - The rate of production delays deterioration of quality. Wherein when the concentration of epsilon-polylysine is 0.06%, the sensory score of fresh radix codonopsis pilosulae at the end of storage is highest, the weight loss rate is lowest, and H 2 O 2 Content sum O 2 - The production rate is the lowest and the fresh-keeping effect is the best.
4.3 influence of storage temperature on fresh-keeping Effect of fresh Sichuan radix Codonopsis
FIG. 3 shows the effect of storage temperature on the fresh keeping effect of fresh Sichuan radix Codonopsis.
As can be seen from fig. 3, the sensory quality of the fresh szechwon tangshen can be remarkably maintained when the storage temperature is between-12 and 20 ℃; significantly reduce the weight loss rate and O 2 - The rate of production delays deterioration of quality. Wherein the fresh radix Codonopsis Lanceolatae has highest sensory score at the end of storage at 4deg.C, lowest weight loss rate, and H 2 O 2 Content sum O 2 - The production rate is low, and the fresh-keeping effect is best.
In summary, the optimal treatment concentration and the optimal storage temperature of the two antistaling agents 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-12-20deg.C, the storage time of fresh radix Codonopsis Lanceolatae can be remarkably prolonged, and the first-stage fresh radix Codonopsis Lanceolatae is still the second-stage better radix Codonopsis Lanceolatae at the end of storage.
2. Response surface experiment
1. Experimental materials
And the same as that of the single factor experiment.
2. Fresh-keeping method
On the basis of 3 single-factor experiments, a response surface scheme is designed by using Design-Expert 11 software. The storage temperature (A), nisin concentration (B) and epsilon-polylysine concentration (C) are respectively 3 independent variables, and the weight coefficient of each index is determined by adopting a analytic hierarchy process-a weight determination method (AHP-CRITIC) based on index correlation to obtain sensory scores, weight loss rate, effective component indexes (codonopsis pilosula alkyne glycoside, syringin and atractylolide III), antioxidant activity indexes (total flavone, total phenol and DPPH free radical clearance rate) and aging-related physiological indexes (H) 2 O 2 Content, & O 2 - Production rate, malondialdehyde content, superoxide dismutase, catalase), browning related physiological indexes (polyphenol oxidase, peroxidase), softening related physiological indexes (pectin methylesterase, polygalacturonase, cellulase, beta-glucosidase) and taking the comprehensive score of 19 evaluation indexes as a response value. The optimal value in each single factor experiment was chosen to be zero level and 17 response surface experiments were performed in combination. The center and the level of the experimental design are shown in table 3, and the specific grouping conditions and the corresponding comprehensive scores of each treatment combination are shown in table 4.
TABLE 3 independent variables and encoded values in response surface analysis experiments
Table 4 response surface experimental grouping and composite scoring
Dividing the pre-cooled fresh radix codonopsis pilosulae into 6 groups at random, immersing the groups in the mixed preservative for 2min, wherein the feed-liquid ratio is 6:5, draining, weighing, putting into a PA/PE microporous fresh-keeping bag, extruding air in the bag, sealing, storing at a corresponding temperature, sampling at 80d in the storage period, measuring the 19 indexes, and repeating each treatment for 3 times.
The preparation method of the mixed preservative comprises the following steps: and calculating and weighing nisin and epsilon-polylysine reagent according to the mass ratio of the feed liquid, dissolving the nisin and epsilon-polylysine reagent in water, and uniformly stirring for later use.
3. Index measurement
3.1 sensory scoring
And the same as that of the single factor experiment.
3.2 weight loss ratio
And the same as that of the single factor experiment.
3.3 H 2 O 2 Content of
And the same as that of the single factor experiment.
3.4 ·O 2 - Rate of production
And the same as that of the single factor experiment.
3.5 cell wall degrading enzyme Activity assay
Pectin Methylesterase (PME), polygalacturonase (PG), cellulase (Cx) and beta-glucosidase (beta-glu) activity determination is referred to Cao Jiankang, guidelines for physiological and biochemical experiments after fruit and vegetable harvest. The glucose amount is taken as the abscissa (X), the A540 nm value is taken as the ordinate (Y), and the standard curve Y=0.6335X-0.0068 (R 2 ＝0.9992)。
The PG enzyme activity unit is 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). The PME enzyme activity unit is 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). The Cx 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). The β -glu enzyme activity unit is defined as the amount of enzyme required to release 1mg of reducing sugar per 1h per 1g fresh weight as one enzyme activity unit (U/g FW).
3.6 other physiological index determination
Malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, catalase (CAT) activity, polyphenol oxidase (PPO) activity and Peroxidase (POD) activity are determined by referring to the detection kit instructions.
3.7 determination of the content of the components of Codonopsis pilosula alkyne glycoside, syringin and atractylenolide III
3.7.1 preparation of sample solution
Taking 4g of fresh szechuan pilose asiabell root powder, precisely weighing, placing into a conical flask with a plug, adding 50mL of methanol, carrying out ultrasonic extraction for 45min, cooling, filtering, concentrating in water bath until the powder is dried, then redissolving the powder into 2mL by using methanol, shaking uniformly, and filtering by using a 0.45 mu m filter membrane to obtain a sample stock solution to be detected. And (5) respectively measuring atractylenolide III and syringin by sample injection according to chromatographic conditions. Precisely sucking 200 mu L of stock solution of the sample to be detected, adding methanol to dilute the stock solution to a volume of 1mL, shaking the stock solution uniformly, filtering the stock solution with a 0.45 mu m filter membrane to obtain the sample to be detected, and sampling the sample to detect the campanumoside according to chromatographic conditions.
3.7.2 drawing of a Standard Curve
Standard curve of codonopsis pilosula acetylenic glycoside: the campanumoside is used as a reference substance to prepare a campanumoside stock solution with the concentration of 1.042 mg/ml. It was then diluted with methanol to a series of gradient codonopsis pilosula acetylenic glycoside control solutions of 32.56, 65.13, 130.25, 260.50, 521.00, 1042.00 μg/mL. Chromatographic conditions: volume ratio acetonitrile: water = 26:74, flow Rate 1.0mL min -1 The detection wavelength is 267nm, the column temperature is 30 ℃, the inlet sample is 10 mu L, the concentration of the reference substance is taken as the abscissa (X), the peak area is taken as the ordinate (Y), and the regression equation is drawn as 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, 26Syringin control solutions with serial gradients of 10, 52.20 and 104.40 mug/mL. The chromatographic conditions are as follows: the mobile phase is acetonitrile (A) -acetic acid water solution (B) with the volume percentage of 0.2 percent, and the gradient elution is as follows: 0-10 min, volume percentage 10% (A); 10-20 min, 15% (A) by volume percent; 20-30 min, volume percentage 10% (A), flow rate 1.0 mL-min -1 The detection wavelength is 220nm, the column temperature is 30 ℃, the inlet sample is 20 mu L, the concentration of the reference substance is taken as the abscissa (X), the peak area is taken as the ordinate (Y), and the regression equation is drawn as Y=90.613X+75.282 (R) 2 ＝1)。
Standard curve for atractylis lactone iii: the atractylenolide III is used as a reference substance to prepare atractylenolide III stock solution with the concentration of 1.238 mg/ml. Then the samples were diluted with methanol to obtain a gradient of 0.31, 1.24, 6.19, 30.95, 309.50, 619.00, 1238.00. Mu.g/mL of the atractylodis lactone III control solution. The chromatographic conditions are acetonitrile in volume ratio: water=71: 29, flow rate 1.0mL min -1 The detection wavelength is 220nm, the column temperature is 30 ℃, the input sample is 20 mu L, the concentration of the reference substance is taken as the abscissa (X), the peak area is taken as the ordinate (Y), and the regression equation is drawn as Y=43.536X+18.169 (R) 2 ＝0.9999)。
3.8 determination of Total Flavonoids, total phenol content and DPPH free radical scavenging ability
3.8.1 preparation of sample solution
Precisely weighing 2g of fresh radix codonopsis pilosulae freeze-dried powder, placing the powder in a 150mL conical flask with a plug, carrying out ultrasonic extraction for 2 times by taking 25mL of 70% ethanol as a solvent for 30min each time, carrying out suction filtration on the filtrate into a 100mL volumetric flask, fixing the volume to a scale, shaking uniformly, and filtering by using a microporous filter membrane to obtain a subsequent filtrate, namely a sample solution.
Drawing of 3.8.2 standard curve
Total flavone standard curve: precisely weighing rutin standard substance 2mg dried to constant weight at 105deg.C, dissolving with appropriate amount of absolute ethanol, fixing volume with absolute ethanol to 10mL, shaking to obtain rutin standard substance stock solution with concentration of 0.2000g/L, and storing at 4deg.C for use. Precisely measuring rutin reference substance solution 0.4,0.8,1.2,1.6,2.0,2.4mL respectively in 10mL measuring flask, adding water to 2.4mL respectively, shaking uniformly, standing for 6min, and adding nitric acid with weight percentage of 5% and sodium nitrite solution 0.4mLShaking up 0.4mL of aluminum, placing for 6min, adding water to the scale after weighing 4mL of sodium hydroxide solution with the weight percentage of 4%, shaking up, and placing for 15min to obtain reference substance solutions with rutin concentrations of 0.008,0.016,0.024,0.032,0.040 and 0.048g/L in sequence. The A505 nm value is taken as an ordinate (Y), the concentration is taken as an abscissa (X), and the standard curve is obtained as Y= 9.6571X-0.0164, (R) 2 ＝0.9991)。
Total phenol standard curve: accurately weighing 1mg of gallic acid standard substance, dissolving with water, fixing volume to 10mL, shaking, diluting 1mL with distilled water by 8 times, and making into 0.0125g/L reference substance standard solution. Accurately sucking 0.8,1.6,2.4,3.2,4.0 and 4.8mL of standard substance solution, respectively placing into 10mL measuring flask, respectively adding distilled water to 2.0mL, mixing, adding 1mL of Fu reagent, and weighing 10% Na after 3min 2 CO 3 2mL of the solution is diluted to 10mL by water, and after heating in a water bath at 30 ℃ for 30min, the solution is taken out and uniformly shaken to prepare the reference substance solutions with the gallic acid concentration of 0.0010,0.0020,0.0030,0.004,0.005 and 0.006g/L in sequence. The concentration is on the abscissa (X) with the value of A765 nm on the ordinate (Y), the standard curve is Y=217.31X+0.0761, (R) 2 ＝0.9991)。
3.8.3 sample assay
Determination of total flavone content: 2mL of the sample solution is precisely measured, the operation is carried out according to the standard curve item, the absorbance is measured, and the total flavone content in the sample is calculated according to the standard curve.
Determination of total phenol content: precisely measuring 1.5mL of the sample solution, operating according to a standard curve item, measuring absorbance, and calculating the total phenol content in the sample according to the standard curve.
DPPH radical scavenging Rate determination: taking DPPH 4mg, placing into a 100mL measuring flask, and fixing the volume to the scale by absolute ethyl alcohol to obtain 0.1mmol/L stock solution, and placing at 4 ℃ for standby. 100 mu L of the sample solution and 2mL of the stock solution are removed, placed in the same test tube, shaken well, reacted in a dark room for 30min, and the absorbance value As is measured at 517nm wavelength. 2mL of the stock solution and 100. Mu.L of absolute ethyl alcohol were removed, placed in the same test tube, shaken well, and the absorbance Ab of the mixed solution was measured. Transferring 2mL of absolute ethyl alcohol and 100 mu L of a test solution, placing the absolute ethyl alcohol and the test solution in the same test tube, measuring absorbance Ac of the mixed solution, and calculating the clearance, wherein the formula is as (3):
3.9 data analysis
And the same as that of the single factor experiment.
4. Analysis of results
4.1 AHP-CRITIC mixed weighting method for determining weight of each index
According to the Box-Behnken experimental Design principle, design Expert 11 software is used for carrying out three-factor three-level response surface experimental Design by taking temperature, nisin concentration and epsilon-polylysine concentration as experimental factors, 17 treatment combinations are obtained, fresh szechuan pilose asiabell roots are subjected to corresponding fresh-keeping treatment and are stored, sampling is carried out at 80d of a storage period, 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 scores, weight loss rates, active ingredients and antioxidant Activity of fresh Sichuan radix Codonopsis
TABLE 6 influence of different fresh-keeping treatments on the physiological index of oxidation aging and browning of fresh Sichuan radix Codonopsis
TABLE 7 influence of different fresh-keeping treatments on the softening physiological index of fresh Sichuan radix Codonopsis
The 19 index weights are determined by adopting an AHP-CRITIC mixed weighting method, and the comprehensive weight calculation formula of the mixed weighting method is as follows (4):
and processing data according to SPSS 22.0 software, and calculating 19 index comprehensive weights according to a formula, wherein the comprehensive weights are shown in Table 8.
TABLE 8 weight analysis results
And (3) carrying out comprehensive scoring comparison on the processed experimental data by respectively adopting the weight coefficients calculated by the AHP, CRITIC and AHP-CRITIC mixed weighting method. The correlation coefficients of the CRITIC and AHP, CRITIC and AHP-CRITIC mixed weighting method and the AHP-CRITIC mixed weighting method are 0.926,0.928,0.998 respectively, the correlation of the three is extremely remarkable (P < 0.01), and the scoring results of the 3 methods are consistent.
From weight coefficient analysis, CRITIC and AHP have a correlation coefficient of-0.038, and the correlation is not significant (p=0.878 > 0.05), indicating that the information reflected by both has no superimposability.
The AHP-CRITIC mixed weighting method is more comprehensive in reflected information and more scientific and reasonable than a single method in terms of both subjective and objective aspects.
4.2 establishment of regression model and analysis of variance
After 17 sets of treatment composite scores (see Table 4 above), regression fits were performed using Design-Expert 11 software. A quadratic polynomial fit equation of the independent variable storage temperature (A), nisin concentration (B) and epsilon-polylysine concentration (C) with the dependent variable composite score (Y) was obtained.
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 analysis of variance results are shown in Table 9.
TABLE 9 response surface analysis of variance results
Note that: p (P)<0.01 is extremely remarkable, P<0.05 is significant: r is R 2 ＝0.9572，R 2 Adj ＝0.9022，CV＝12.26％。
From the data results in table 9, the model regression equation is obvious, the mismatch term is not obvious, and the fitting degree of the regression equation is good, so that the method has statistical significance. Determining the coefficient R 2 0.9572, the measured value is closer to the predicted value. Correction coefficient R 2 Adj 0.9022, the three factors have 90.22% influence on the fresh-keeping effect of the fresh radix codonopsis pilosulae, and the experiment is proved to have higher accuracy. The coefficient of variation CV was 12.26% and less than 15%, indicating good stability of the experiment.
According to the value of P, the quality guarantee period of the fresh radix codonopsis pilosulae is influenced by all factors as follows: a > C > B, and quadratic term A 2 、B 2 And C 2 The impact on the composite score is very pronounced.
4.3 two-factor interaction response surface analysis
The response curve diagram and the contour diagram of the influence of 3 interaction items are shown in fig. 4-9, which show that the level selection of each experimental factor is reasonable, wherein the influence of the interaction of temperature and two kinds of antistaling agents on the comprehensive score of the storage end stage of the fresh radix codonopsis pilosulae is strong, and the influence of the interaction between the two kinds of antistaling agents on the comprehensive score is secondary, and the result is consistent with the result of the value P.
4.4 verification of optimal conditions
The optimal method for obtaining the storage and fresh-keeping of the codonopsis pilosula after picking by using Design-Expert 11 software comprises the following steps: the mixed preservative contains 0.05% of nisin and 0.05% of epsilon-polylysine, and the storage temperature is 2 ℃. Under the condition, three verification experiments are carried out, the 19 indexes are measured on the 80 th day of storage, the results are shown in table 10, comprehensive scores are obtained by calculation according to AHP-CRITIC mixing weights in table 8, the comprehensive scores are 66.8810, 63.5256 and 60.7902 respectively, the average value is 63.7323 and is close to a model predicted value 62.4208, the fitting degree of the equation is good, the reliability of the obtained optimal fresh-keeping method is high, and the method can be used for industrialized fresh-keeping storage of fresh radix codonopsis pilosulae.
Table 10 validates the results of the experiment
Example 2
A method for storing and preserving radix codonopsis pilosulae after picking comprises the following steps:
(1) Immersing the pre-cooled fresh szechwon tangshen root into the mixed preservative for 2min, wherein the feed-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 pilosulae, weighing, placing into a PA/PE microporous fresh-keeping bag, extruding air in the bag, sealing, and storing at 2 ℃.
Example 3
And (3) under the optimal fresh-keeping condition (namely, fresh keeping is carried out by adopting the method of the example 2), and measuring the volatile compounds at the end of the storage period of the fresh szechwon tangshen root.
1. Material
Fresh radix codonopsis pilosulae sample (S0); storing the 40 d-th radix codonopsis pilosulae sample (LPN 40) under the optimal fresh-keeping condition; storing the 80d Sichuan radix codonopsis sample (LPN 80) under the optimal fresh-keeping condition; the 3d Szechwan radix Codonopsis sample (RT 3) and the 6d Szechwan radix Codonopsis sample (RT 6) were stored at room temperature.
2. Method of
2.1 instruments
Flavour Flavor analyzer (GAS-ion mobility spectrometry), GAS company, germany.
2.2 sample handling
1g of the sample is 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 injection volume is 500 mu L, the incubation time is 15min, the incubation temperature is 60 ℃, and the analysis time is 20min. 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 flow rate: 150mL/min; carrier gas flow rate: 2mL/min (0-2 min), 2-10 mL/min (2-10 min), 10-100 mL/min (10-20 min); ion transfer tube temperature: 45 ℃; sample injection needle temperature: 85 ℃.
3. Analysis of results
The fingerprints of the volatile substances of the codonopsis pilosula with 5 different storage periods are shown in figure 10, and 102 volatile organic compounds are detected, wherein 9 esters, 23 aldehydes, 16 alcohols, 9 ketones, 1 acid, 1 thiophene, 1 furan and 42 unknown compounds.
Of the quantitative volatiles, the aldehydes and alcohols content was the greatest. The main volatile matters of the fresh szechwon tangshen root are n-hexanal, (E) -2-hexenal and the like, wherein the n-hexanal is the main component of the special fragrance. From the results, the storage preservation method shows that the content of hexanal gradually decreases, and the preservation method in example 2 can maintain good fragrance quality of radix codonopsis to a certain extent.
Fig. 10 is a graph (fingerprint) of volatile matter gallry Plot of radix codonopsis pilosulae with different storage periods.
Consistent with the conclusion of the Gallery Plot, the PCA analysis can distinguish between 5 samples from fig. 11. Wherein LPN40, LPN80 are closer to S0 and the overall volatile components are similar. It is further shown that the preservation method of example 2 can better retain the volatile flavor of fresh radix codonopsis pilosulae during storage.
Fig. 11 is a PCA diagram of volatile materials of Sichuan radix Codonopsis with different storage periods.
Example 4
Compared with the embodiment 2, the concentration of nisin in the mixed preservative is 0.06%, the concentration of epsilon-polylysine is 0.06%, and the preservative is stored at 4 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 5
Compared with the embodiment 2, the concentration of nisin in the mixed preservative is 0.02%, the concentration of epsilon-polylysine is 0.06%, and the preservative is stored at 4 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 6
Compared with the embodiment 2, the concentration of nisin in the mixed preservative is 0.02%, the concentration of epsilon-polylysine is 0.05%, and the preservative is stored at 2 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 7
Compared with the embodiment 2, the concentration of nisin in the mixed preservative is 0.08%, the concentration of epsilon-polylysine is 0.06%, and the preservative is stored at 0 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 8
Compared with the embodiment 2, the concentration of nisin in the preservative is 0.08%, and the preservative is stored at 2 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 9
Compared with the embodiment 2, the concentration of nisin in the preservative is 0.04%, and the preservative is stored at-4 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 10
Compared with the embodiment 2, the concentration of epsilon-polylysine in the preservative is 0.06%, and the preservative is stored at 0 ℃ after being packaged by a PA/PE microporous preservative bag.
Example 11
Compared with the embodiment 2, the concentration of epsilon-polylysine in the preservative is 0.04%, and the preservative is stored at 12 ℃ after being packaged by a PA/PE microporous preservative bag.
Finally, it should be noted that: the foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A method for preserving radix codonopsis pilosulae after picking is characterized in that: soaking precooled fresh radix codonopsis pilosulae in preservative, draining, placing into PA/PE microporous preservative bag, extruding air in the bag, sealing, and storing at-12-20deg.C; the preservative is aqueous solution of nisin and/or epsilon-polylysine;
in the preservative, the weight percentage concentration of nisin is 0.02-0.10%; and/or
In the preservative, the weight percentage concentration of epsilon-polylysine is 0.02-0.10%;
the pre-cooling is pre-cooling at 0+/-1 ℃;
the soaking time is 1.5-2.5min.
2. The method according to claim 1, characterized in that: 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%.
3. The method according to claim 1 or 2, characterized in that: the preservative is an aqueous solution of nisin and epsilon-polylysine.
4. A method according to claim 3, characterized in that: the preservative consists of the following components in percentage by weight: nisin 0.05%, epsilon-polylysine 0.05% and water balance.
5. The method according to claim 1, characterized in that: sealing and storing at-4-12deg.C.
6. The method according to claim 5, wherein: sealing and storing at 2 ℃.
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