CN114014946B - Preparation method of yellow tea polysaccharide and application of yellow tea polysaccharide prepared by preparation method in preserved meat - Google Patents
Preparation method of yellow tea polysaccharide and application of yellow tea polysaccharide prepared by preparation method in preserved meat Download PDFInfo
- Publication number
- CN114014946B CN114014946B CN202210010856.1A CN202210010856A CN114014946B CN 114014946 B CN114014946 B CN 114014946B CN 202210010856 A CN202210010856 A CN 202210010856A CN 114014946 B CN114014946 B CN 114014946B
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- China
- Prior art keywords
- yellow tea
- tea polysaccharide
- polysaccharide composition
- yellow
- polysaccharide
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Images
Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
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- 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
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
- A23L13/42—Additives other than enzymes or microorganisms in meat products or meat meals
- A23L13/428—Addition of flavours, spices, colours, amino acids or their salts, peptides, vitamins, yeast extract or autolysate, nucleic acid or derivatives, organic acidifying agents or their salts or acidogens, sweeteners, e.g. sugars or sugar alcohols; Addition of alcohol-containing products
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- 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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/40—Colouring or decolouring of foods
- A23L5/41—Retaining or modifying natural colour by use of additives, e.g. optical brighteners
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention discloses a method for preparing yellow tea polysaccharide by an ultrahigh pressure technology and application of the prepared yellow tea polysaccharide in preserved meat. The preparation method mainly comprises the following steps: yellow tea → washing → drying at 60 deg.C → crushing → sieving with 100 mesh sieve → weighing a proper amount of dry powder of yellow tea → ultra-high pressure extraction → centrifugation for taking supernatant → concentration → precipitation → washing → redissolution → deproteinization → collection of supernatant → dialysis → freeze drying → obtaining yellow tea polysaccharide powder. The yellow tea polysaccharide is used for pickling the preserved meat, and has good antioxidant activity, bacteriostatic activity and color protection characteristics.
Description
Technical Field
The invention relates to preparation of yellow tea polysaccharide, in particular to yellow tea polysaccharide prepared by an ultrahigh pressure technology and application thereof in preserved meat.
Background
Tea is one of three major beverages in the world, and has a history of over 5000 years in dietary and medicinal aspects. Tea leaves contain a variety of active ingredients including polysaccharides, polyphenols, caffeine, amino acids, carbohydrates, proteins, lipids and trace amounts of other substances. Yellow tea is one of six major tea types in China, belongs to light fermented tea, and is known for its unique 'stuffy yellow' process. The yellow tea polysaccharide is an important functional macromolecular substance in tea, contains uronic acid, polyphenol, pectin, protein, mineral elements and the like, has various biological activities, and has more and more attention on health care efficacy along with wide application of the tea in the aspects of medicine, health care, food processing and the like. The current research shows that the yellow tea polysaccharide has great prospects in the aspects of regulating immunity, resisting tumors, resisting thrombus, reducing blood fat and blood sugar, resisting oxidation and the like, and has special efficacy for treating diabetes.
Although the yellow tea polysaccharide has so many pharmacological effects, people can only take the tea polysaccharide by drinking tea because the extraction technology of the yellow tea polysaccharide is still relatively backward, but the quantity of the tea polysaccharide taken in is very small, and the good pharmacological effect is difficult to achieve. The traditional preparation process is that tea leaves are crushed, firstly leached by acidic, neutral or alkalescent water at a certain temperature, and then the crude product is prepared by precipitation separation or column chromatography separation. The traditional extraction process has the disadvantages of more flows, high difficulty, high cost, poor oxidation resistance of the extracted tea polysaccharide, poor method reproducibility and low yield of the yellow tea polysaccharide. In addition, because the polarity of tea polyphenol and tea polysaccharide has a certain difference, it is difficult to find a solvent capable of efficiently and synchronously extracting tea polyphenol and tea polysaccharide. In addition, the raw materials used in the existing ultrahigh pressure mode (CN 112150182A) are common selenium-rich tea, the process optimization degree is low, the yield of the selenium polysaccharide is as high as 2.48%, the sugar yield is not high, the selenium polysaccharide is mainly used as a component of a functional product formula and plays roles of reducing blood sugar, reducing blood fat and supplementing selenium, the components of the obtained selenium polysaccharide are not further analyzed, and other physiological activities of the selenium-rich polysaccharide are not clear.
The research aims to optimize the extraction method of the tea polysaccharide through an ultrahigh pressure technology, greatly reduces the extraction time, increases the extraction efficiency and yield of the tea polysaccharide and saves the extraction cost compared with the traditional ultrahigh pressure extraction mode. Meanwhile, the ultrahigh pressure technology as a non-thermal processing extraction method can reduce the loss and damage of heat sensitive components. In addition, the raw material in the research is yellow tea or yellow tea old tea, and the extraction of the yellow tea polysaccharide from the raw material by the ultrahigh pressure technology belongs to the optimization of low-value resources, so that the comprehensive utilization value of the yellow tea old tea is favorably improved, and the pharmacological effect of the yellow tea polysaccharide is better exerted. More significantly, through optimization of a solvent, parameters, a process and the like, the extraction efficiency of the tea polysaccharide is improved, a certain amount of tea polyphenol can be obtained, the synchronous extraction of the tea polysaccharide and the tea polyphenol is realized, and a certain proportion of tea polyphenol exists in a final crude polysaccharide product. Therefore, the optimization of the extraction process of the yellow tea polysaccharide in the research is of great significance.
The preserved meat has a long history in China, is one of the traditional dry-cured meat products, and is mainly characterized in that the finished product has beautiful color, rich flavor, better storability and convenient eating. The fat oxidation product is also an important characteristic flavor substance component of the traditional preserved meat product in China, and the flavor components are required to be reserved for keeping the traditional characteristic flavor. Medical research shows that the fat oxidation products can induce various chronic diseases of the body and are the main causes of human body aging and cardiovascular diseases. Therefore, the development of natural antioxidants without toxic and side effects has great significance, and at present, more researches have found that some plant antioxidant active ingredients can effectively inhibit the oxidation of lipid and protein of meat and simultaneously reduce nitrite in preserved meat.
The invention applies the crude yellow tea polysaccharide to the preserved meat product, and the tea polysaccharide mainly comprises 26.1 percent of neutral sugar, 35.09 percent of uronic acid, 10 percent of tea polyphenol and 13.8 percent of protein. Wherein the neutral sugar comprises 8 sugars of mannose, rhamnose, ribose, glucose, galactose, xylose, arabinose and fucose, and the molar percentage is as follows: 20.46% mannose, 11.61% rhamnose, 3.19% ribose, 20.97% glucose, 17.14% galactose, 8.02% xylose, 15.85% arabinose, 2.76% fucose. The proper proportion has special properties, and the antioxidant activity, the bacteriostatic activity and the color protection characteristic are good. The antibacterial property, the oxidation resistance, the biodegradability and the color protection property of the preserved meat can be fully exerted in the pickling process of the preserved meat, the quality of the preserved meat is improved, the content of nitrite in the preserved meat is reduced, the flavor substance components in the preserved meat are enriched, and meanwhile, a part of nutrition and health care functions can be endowed to the preserved meat.
At present, the traditional preserved meat pickling method mostly adopts a dry pickling method. However, the traditional dry pickling method has the defects of slow salt penetration speed and uneven penetration amount, so that the texture of meat is influenced, the color and the taste of a cured meat finished product are inconsistent, and the requirement of large-scale industrial production cannot be met. Compared with the traditional preparation method of the tea-flavored preserved meat (CN 105379823A), the rolling and kneading pickling method can accelerate the pickling speed, shorten the pickling time, increase the water retention of meat products and improve the product quality. The vacuum rolling and kneading technology is combined with rolling and kneading under the vacuum condition, so that the permeation rate of the curing agent can be improved, the flavor of tea aroma and the tenderness of meat quality can be increased, and the preserved meat is endowed with unique flavor. In addition, tea leaves are mostly used as a pickling material in the traditional tea-flavor preserved meat preparation method, and the yellow tea polysaccharide extracted by using the preparation method disclosed by the invention is added to be matched with a vacuum rolling pickling method, so that the antibacterial property, the oxidation resistance and the color protection property of the yellow tea polysaccharide can be fully utilized, the preserved meat with good sensory properties can be pickled even if nitrite is not added, and the nitre-free tea-flavor preserved meat is produced.
Disclosure of Invention
The invention aims to effectively extract and separate polysaccharides in yellow tea by an ultrahigh pressure technology, optimize the extraction process of the yellow tea polysaccharides, avoid the problems of high extraction temperature, long extraction time, large extraction solvent amount, low tea polysaccharide yield and the like of the traditional yellow tea polysaccharides, and provide a method for efficiently extracting the yellow tea polysaccharides by the ultrahigh pressure technology. The yellow tea polysaccharide extracted by the method has unique proportion of neutral sugar, tea polyphenol, uronic acid and protein and good antioxidant activity, and when the yellow tea polysaccharide is applied to the quality improvement of preserved meat, the antibacterial property, the antioxidant property and the color protection property of the yellow tea polysaccharide can be fully utilized, the nitrite in the preserved meat can be obviously reduced, the flavor substance components in the preserved meat can be enriched, and meanwhile, the nutritional and health-care functions of a part of the yellow tea polysaccharide can be endowed to the preserved meat. In addition, in the cured meat pickling process, the cured meat with good sensory properties can be pickled by the tea polysaccharide adding process and the vacuum tumbling pickling method even if nitrite is not added.
The purpose of the invention is realized by the following technical scheme:
the preparation method of the yellow tea polysaccharide is characterized by comprising the following steps:
step 1) yellow tea pretreatment: cleaning old yellow tea, oven drying, pulverizing, sieving, making into yellow tea dry powder, sealing, drying and storing;
step 2) weighing a proper amount of yellow tea dry powder, and performing ultrahigh pressure extraction through a proper amount of solvent;
step 3) centrifuging the extraction solution for 5-15min at 7000r/min of 5000-;
step 4), concentrating the supernatant to 10% -20% of the original volume, adding 90% ethanol solution with 4-6 times of volume into the concentrated solution, and precipitating for 24 h;
step 5) collecting the precipitate, washing the precipitate for 2-3 times by using absolute ethyl alcohol and diethyl ether respectively, and redissolving the washed precipitate in water;
step 6), re-dissolving the washed precipitate in water, mixing the re-dissolved precipitate with Sevage solvent according to a volume ratio of 3:1, removing protein, collecting supernatant, and collecting for 2 times;
step 7), combining the two supernatants, dialyzing the supernatant by using an 8kDa dialysis bag, and freeze-drying to obtain the yellow tea crude polysaccharide;
in order to further realize the aim of the invention, preferably, the drying after the yellow tea is cleaned is blast drying, the adopted drying temperature is 60-80 ℃, and the yellow tea is crushed and then sieved by a sieve of 90-100 meshes;
preferably, the extraction solution is centrifuged at 6000r/min for 5 min;
preferably, the supernatant is concentrated to 15% of the original volume;
preferably, a 5-fold volume of 90% ethanol solution is added to the concentrate and allowed to settle for 24 h.
Preferably, the extraction solvent in step 2) is distilled water,
preferably, the extraction time in step 2) is 3-15 minutes;
preferably, the extraction pressure in the step 2) is 200-600 MPa;
preferably, the feed-liquid ratio in the step 2) is 1:5-25 g/mL;
preferably, the Sevage solvent in the step 6) is a mixture of chloroform and n-butanol with a volume ratio of 4: 1;
preferably, the extraction conditions of the yellow tea polysaccharide are as follows: the feed-liquid ratio is 20g/mL, the extraction time is 6min, and the extraction pressure is 300 MPa.
Preferably, the yield of the yellow tea polysaccharide is 9.32-10.81%, and the yellow tea polysaccharide mainly comprises 26.1-28.9% of neutral sugar, 35.09-38.42% of uronic acid, 10-12.5% of polyphenol, 3.8-4.2% of protein and the like. The neutral sugar of the yellow tea polysaccharide consists of 8 sugars, namely mannose, rhamnose, ribose, glucose, galactose, xylose, arabinose and fucose, and the molar percentage is 20.46 to 23.12 percent, 8.37 to 11.61 percent, 3.19 to 4.52 percent, 17.54 to 20.97 percent, 15.59 to 17.14 percent, 8.02 to 9.35 percent, 13.49 to 15.85 percent and 2.76 to 3.63 percent.
Preferably, the yield of the yellow tea polysaccharide is 10.81%, and the yellow tea polysaccharide has good in-vitro antioxidant activity which can reach 16.82 +/-0.28U mL-1. The yellow tea polysaccharide mainly contains neutral sugar 26.1%, uronic acid 35.09%, polyphenol 10%, and protein 3.8%. The composition of the neutral sugar comprises the following components in percentage by mole: 20.46% mannose, 11.61% rhamnose, 3.19% ribose, 20.97% glucose, 17.14% galactose, 8.02% xylose, 15.85% arabinose, 2.76% fucose.
Compared with the traditional yellow tea polysaccharide extraction process, the method can obviously improve the yield of the yellow tea polysaccharide. The extraction method of the ultrahigh pressure technology used by the invention not only has the advantages of low extraction temperature and short extraction time, but also has the advantages of high efficiency, good repeatability, maximum retention of active ingredients and the like, and can obtain yellow tea polysaccharide with higher antioxidant activity; the method has simple and convenient extraction technology operation, can retain the physiological activity of the yellow tea polysaccharide to a greater extent, meets the requirements of consumers on the flavor and the efficacy of the product, and has wide market prospect.
The yellow tea polysaccharide of 1.0g/kg is applied to the cured meat pickling process, and meanwhile, the vacuum rolling pickling method is adopted, so that compared with a blank group, the addition of the yellow tea polysaccharide in the cured meat can better improve the physicochemical properties and flavor of the product, and improve the quality of the cured meat. The concrete expression is as follows: after the tea polysaccharide is added, the sample keeps good color, the POV value and the TBARs value are obviously reduced, meanwhile, nitrite residue is hardly generated, and the tea polysaccharide has good sensory quality, good economic value and good use prospect.
Drawings
FIG. 1 is a graph showing the effect of extraction time on polysaccharide yield
FIG. 2 Effect of feed-liquid ratio on polysaccharide yield
FIG. 3 Effect of extraction pressure on polysaccharide yield
FIG. 4 SOD activity of the product of example 1
FIG. 5 monosaccharide composition analysis of the product of example 1
Figure 6 LDA analysis of different groups of bacon flavors.
Detailed Description
The present invention will be described in further detail with reference to examples, but the present invention is not limited to these examples.
Example 1 preparation of yellow tea polysaccharide
The method comprises the following steps:
step 1) tea leaf pretreatment: cleaning old yellow tea, drying at 60 deg.C, pulverizing, sieving with 100 mesh sieve to obtain yellow tea dry powder, sealing, drying and storing;
step 2) weighing 5g of yellow tea dry powder, and carrying out ultrahigh pressure extraction by using distilled water, wherein the extraction time is 6 minutes, the extraction pressure is 300MPa, and the material-liquid ratio is 20 g/mL;
step 3) centrifuging the extraction solution for 15min at 5000r/min, and collecting supernatant;
step 4), concentrating the supernatant to 10% of the original volume, adding 90% ethanol solution with 4 times of volume into the concentrated solution, and precipitating for 24 h;
step 5) collecting the precipitate, washing the precipitate for 3 times by using absolute ethyl alcohol and diethyl ether respectively, and redissolving the washed precipitate in water;
step 6), re-dissolving the washed precipitate in water, mixing the re-dissolved precipitate with Sevage solvent according to a volume ratio of 3:1, removing protein, collecting supernatant, and collecting for 2 times;
and 7) combining the two supernatants, dialyzing the supernatant by using an 8kDa dialysis bag, and freeze-drying to obtain the yellow tea crude polysaccharide.
Example 2 preparation of yellow tea polysaccharide
The method comprises the following steps:
step 1) tea leaf pretreatment: cleaning old yellow tea, drying at 60 deg.C, pulverizing, sieving with 100 mesh sieve to obtain yellow tea dry powder, sealing, drying and storing;
step 2) weighing 5g of yellow tea dry powder, and carrying out ultrahigh pressure extraction by using distilled water, wherein the extraction time is respectively 3, 6, 9, 12 and 15 minutes, the extraction pressure is 300MPa, and the material-liquid ratio is 20 g/mL;
step 3) centrifuging the extraction solution for 15min at 5000r/min, and collecting supernatant;
step 4), concentrating the supernatant to 10% of the original volume, adding 90% ethanol solution with 4 times of volume into the concentrated solution, and precipitating for 24 h;
step 5) collecting the precipitate, washing the precipitate for 3 times by using absolute ethyl alcohol and diethyl ether respectively, and redissolving the washed precipitate in water;
step 6), re-dissolving the washed precipitate in water, mixing the re-dissolved precipitate with Sevage solvent according to a volume ratio of 3:1, removing protein, collecting supernatant, and collecting for 2 times;
and 7) combining the two supernatants, dialyzing the supernatant by using an 8kDa dialysis bag, and freeze-drying to obtain the yellow tea crude polysaccharide, wherein the correlation between the extraction time and the polysaccharide yield is shown in figure 1.
Example 3 preparation of yellow tea polysaccharide
The method comprises the following steps:
step 1) tea leaf pretreatment: cleaning old yellow tea, drying at 60 deg.C, pulverizing, sieving with 100 mesh sieve to obtain yellow tea dry powder, sealing, drying and storing;
step 2) weighing 5g of yellow tea dry powder, and carrying out ultrahigh pressure extraction by using distilled water, wherein the extraction time is 6 minutes, the extraction pressures are respectively 200MPa, 300MPa, 400MPa, 500MPa and 600MPa, and the material-liquid ratio is 20 g/mL;
step 3) centrifuging the extraction solution for 15min at 5000r/min, and collecting supernatant;
step 4), concentrating the supernatant to 10% of the original volume, adding 90% ethanol solution with 4 times of volume into the concentrated solution, and precipitating for 24 h;
step 5) collecting the precipitate, washing the precipitate for 3 times by using absolute ethyl alcohol and diethyl ether respectively, and redissolving the washed precipitate in water;
step 6), re-dissolving the washed precipitate in water, mixing the re-dissolved precipitate with Sevage solvent according to a volume ratio of 3:1, removing protein, collecting supernatant, and collecting for 2 times;
and 7) combining the two supernatants, dialyzing the supernatant by using an 8kDa dialysis bag, and freeze-drying to obtain the yellow tea crude polysaccharide, wherein the correlation between the extraction pressure and the polysaccharide yield is shown in figure 3.
Example 4 preparation of yellow tea polysaccharide
The method comprises the following steps:
step 1) tea leaf pretreatment: cleaning old yellow tea, drying at 60 deg.C, pulverizing, sieving with 100 mesh sieve to obtain yellow tea dry powder, sealing, drying and storing;
step 2) weighing 5g of yellow tea dry powder, and carrying out ultrahigh pressure extraction by using distilled water, wherein the extraction time is 6 minutes, the extraction pressure is 300MPa, and the material-liquid ratios are respectively 5, 10, 15, 20 and 25 g/mL;
step 3) centrifuging the extraction solution for 15min at 5000r/min, and collecting supernatant;
step 4), concentrating the supernatant to 10% of the original volume, adding 90% ethanol solution with 4 times of volume into the concentrated solution, and precipitating for 24 h;
step 5) collecting the precipitate, washing the precipitate for 3 times by using absolute ethyl alcohol and diethyl ether respectively, and redissolving the washed precipitate in water;
step 6), re-dissolving the washed precipitate in water, mixing the re-dissolved precipitate with Sevage solvent according to a volume ratio of 3:1, removing protein, collecting supernatant, and collecting for 2 times;
and 7) combining the two supernatants, dialyzing the supernatant by using an 8kDa dialysis bag, and freeze-drying to obtain the yellow tea crude polysaccharide, wherein the correlation between the material-liquid ratio and the polysaccharide yield is shown in figure 2.
Example 5 determination of polysaccharide fraction
Measuring the content of neutral sugar by adopting a phenol-sulfuric acid colorimetric method; measuring the protein content by adopting a Coomassie brilliant blue G-250 method; and measuring the content of uronic acid by adopting a phenol-carbazole colorimetric method, and measuring the content of tea polyphenol by adopting a ferrous tartrate colorimetric method.
The yellow tea polysaccharide mainly comprises neutral sugar 26.1-28.9%, uronic acid 35.09-38.42%, polyphenol 10-12.5%, and protein 3.8-4.2%.
The yellow tea polysaccharide contains neutral sugar and uronic acid as main components, and also contains 3.8-4.2% of protein and 10% -12.5% of polyphenol, which indicates that the yellow tea polysaccharide is an acidic polysaccharide conjugated with protein. The yellow tea polysaccharide after vacuum freeze drying is light brown layered solid, is soluble in water and insoluble in high-concentration organic solvent.
Example 6 yellow tea polysaccharide in vitro antioxidant assay
SOD in the polysaccharide was measured by using the kit method, and the results are shown in FIG. 4. The SOD activity of tea polysaccharide is related to the concentration in dosage, and when the concentration of polysaccharide is 50 μ g mL-1And 200. mu.g mL-1The SOD activity of tea polysaccharide is at a high level when the polysaccharide concentration is 800 μ g mL-1The SOD activity of tea polysaccharide is 16.82 + -0.28U mL-1。
Example 7 analysis of monosaccharide composition of yellow tea polysaccharide
Preparation of a sample: precisely weighing about 5mg of yellow tea polysaccharide freeze-dried powder, adding 2mL of 2mol/L TFA solution, sealing, carrying out oil bath reaction at 120 ℃ for 6h, carrying out rotary evaporation and reduced pressure concentration until the yellow tea polysaccharide freeze-dried powder is dried, adding about 3mL of methanol to dissolve a dried substance, continuously carrying out rotary evaporation to dryness, repeating the operation for 3 times, adding 1mL of deionized water to dissolve the dried substance, adding 200 mu L of PMP-methanol solution and 200 mu L of 0.3mol/L sodium hydroxide solution into a test tube of which the volume is 0.1mL, carrying out water bath reaction at 70 ℃ for 30min, cooling to room temperature (25 +/-2), adding 210 mu L of 0.3mol/L hydrogen chloride solution, extracting with 1mL of chloroform, oscillating uniformly, and centrifuging at 10000r/min for 5 min. After centrifugation, the supernatant was collected and the operation was repeated 3 times. And finally filtering with a 0.45-micron filter membrane for high performance liquid chromatography detection. Chromatographic conditions are as follows: the column was Agilent ZORBAX Eclipse XDB-C18 (4.6 mm. times.250 mm, 5 μm); the mobile phase is 0.1mol/L phosphate buffer solution-acetonitrile (volume ratio is 83: 17); the flow rate is 1 mL/min; the sample injection amount is 20 mu L; the detection wavelength was 250 nm.
The correlation result is shown in fig. 5, the left side is a standard mixed monosaccharide spectrogram, the right side is a yellow tea polysaccharide sample spectrogram, and the peaks in the spectrogram are respectively: 1. mannose (Man); 2. rhamnose (Rha); 3. ribose (Rib); 4. galacturonic acid (GalA); 5. glucose (Glc); 6. galactose (Gal); 7. xylose (Xyl); 8. arabinose (Ara); 9. fucose (Fuc).
The neutral sugar of the yellow tea polysaccharide consists of 8 sugars, namely mannose, rhamnose, ribose, glucose, galactose, xylose, arabinose and fucose, and the molar percentage is 20.46 to 23.12 percent, 8.37 to 11.61 percent, 3.19 to 4.52 percent, 17.54 to 20.97 percent, 15.59 to 17.14 percent, 8.02 to 9.35 percent, 13.49 to 15.85 percent and 2.76 to 3.63 percent. Wherein the contents of mannose, glucose, galactose and arabinose are relatively high, and the contents of ribose and fucose are small.
Example 8 application of yellow tea polysaccharide to quality enhancement of preserved meat (containing nitrite)
The method comprises the steps of adopting a traditional bacon making process, scraping residual hair and dirt on a skin layer with pork back meat with the fat-lean ratio of 3: 7-4: 6, cleaning, trimming and cutting along the edge, cutting into meat blocks with the length of 15-20 cm and the width of 3cm, cutting a hole at the top end, rinsing in warm water at 30 ℃ for 2min, removing floating oil and dirt on the surface of meat strips, taking out and draining water. The pickling adopts a vacuum rolling pickling method, the tea-flavored pickling agent is uniformly coated on the surface of meat, the meat is placed in a vacuum container and is vacuumized by a vacuum pump, the vacuum degree is 86kPa, the meat is uniformly pickled by rolling for 1min every 24h, and the rotating speed is 20 r/min; the pickling temperature is 4-6 ℃, and the pickling time is 5 days. 3.5kg of common salt is used for every 100kg of raw meat. Auxiliary materials: white granulated sugar, soy sauce, white spirit, aniseed, cassia bark, pepper, tea polysaccharide and nitrite. Preheating for 1h at 45 deg.C (humidity of 49%), baking for 12h at 60 deg.C (humidity of 62%), smoking with crushed tea dust and cypress chips for 8h to obtain the final product, and suspending and storing the preserved meat at 20 deg.C for 7d to determine its physicochemical index. Preparing a blank group (A group) and a tea polysaccharide group (1.0 g/kg yellow tea polysaccharide B group) by the same process, and measuring the color difference, the peroxide value (POV), the thiobarbituric acid reactant (TBARs) value, the nitrite residue, the sensory evaluation and the volatile flavor substances of the two groups of bacons.
Example 9 application of yellow tea polysaccharide in quality improvement of preserved meat (without nitrite)
The method comprises the steps of adopting a traditional bacon making process, scraping residual hair and dirt on a skin layer with pork back meat with the fat-lean ratio of 3: 7-4: 6, cleaning, trimming and cutting along the edge, cutting into meat blocks with the length of 15-20 cm and the width of 3cm, cutting a hole at the top end, rinsing in warm water at 30 ℃ for 2min, removing floating oil and dirt on the surface of meat strips, taking out and draining water. The pickling process comprises uniformly coating the pickling agent on the surface of meat, placing into a vacuum container, vacuumizing with a vacuum degree of 86kPa, rolling for 1min every 24h for uniformly pickling, and rotating at a speed of 20 r/min; the pickling temperature is 4-6 ℃, and the pickling time is 5 days. 3.5kg of common salt is used for every 100kg of raw meat. Auxiliary materials: white granulated sugar, soy sauce, white spirit, aniseed, cassia bark, pepper and tea polysaccharide. Preheating for 1h at 45 deg.C (humidity of 49%), baking for 12h at 60 deg.C (humidity of 62%), smoking with crushed tea dust and cypress chips for 8h to obtain the final product, and suspending and storing the preserved meat at 20 deg.C for 7d to determine its physicochemical index. This example is a tea polysaccharide and no nitrite group added (1.0 g/kg yellow tea polysaccharide group C).
Measuring color difference of preserved meat, peroxide value (POV), thiobarbituric acid reactant (TBARs) value, nitrite residue, sensory evaluation, and volatile flavor substance.
The peroxide value was measured by direct titration according to GB 5009.227-2016 (national food safety standards for peroxide value determination) and converted to meq/kg of peroxide/kg of fat.
The TBARs value was measured by spectrophotometry according to GB 5009.181-2016 "measurement of malondialdehyde in food safety Standard".
The determination of the nitrite content is carried out by adopting a spectrophotometric method of GB 5009.33-2016 (determination of nitrite and nitrate in food safety standard food).
Sensory evaluation is based on sensory requirements in GB 2730-2015 national food safety Standard pickled and cured meat product, and fuzzy mathematical sensory evaluation method is applied. An evaluation group consisting of 10 evaluators performed sensory evaluation on 3 indexes of color, smell and state of 3 finished bacons, and scored from 4 grades of poor, general, good and excellent.
TABLE 1 sensory evaluation criteria for cured meat
And (3) carrying out electronic nose analysis, namely chopping the sample, putting 1.0g of the uniform sample into a 15mL headspace sample injection bottle, sealing the sample by using a double-layer preservative film, standing the sample at normal temperature for 30min, carrying out electronic nose detection analysis by using a headspace inspiration method, and repeating each sample for 3 times. The sampling parameters of the electronic nose are set as follows: the sample measurement interval time is 1s, the sensor automatic cleaning time is 60s, the sensor zero setting time is 10s, the sample injection preparation time is 5s, the analysis sampling time is 70s, and the sample injection flow is 400 mL/min. And then extracting characteristic values of each sensor of the electronic nose, and performing Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA) on the data by utilizing Winmaster software matched with a PEN3 sensor. In order to ensure the stability and accuracy of experimental data, 50-52 s of data in the determination process is selected for subsequent analysis.
GC-MS analysis shows that 3.0g of uniform sample is extracted from volatile flavor substances in a 15mL headspace bottle, the headspace bottle is sealed by a polytetrafluoroethylene spacer, a solid phase micro-extractor is inserted into the sample bottle, an extraction head is pushed out, the solid phase micro-extractor is exposed at the upper part of the sample bottle, and extraction is carried out for 40min at 60 ℃. After extraction, the extraction head is inserted into a gas chromatography sample inlet to desorb at 250 ℃ for 6min, and simultaneously an instrument is started to collect data. GC-MS detection conditions and data processing GC conditions: column model SH-Rxi-5 SilMS (30 m × 0.25mm × 0.25 μm), helium as carrier gas, flow rate of 1.19mL/min, injection port temperature of 250 deg.C, no flow diversion, initial temperature of 35 deg.C, holding for 3min, raising to 130 deg.C at 4 deg.C/min and holding for 2min, and raising to 230 deg.C at 8 deg.C/min and holding for 3 min. MS conditions: the ionization mode is EI, the ionization energy is 70eV, the interface temperature is 250 ℃, the ion source temperature is 200 ℃, and the mass scanning range (m/z) is 35-450.
TABLE 2 sensory evaluation results of bacon of different groups
Note: the evaluation grades of the bacon are excellent (V1), good (V2), normal (V3) and poor (V4).
TABLE 3 difference in physicochemical Properties of different groups of bacons
TABLE 4 types and percentages of volatile substances in different cured meat samples
The result shows that the sample keeps better color after the tea polysaccharide is added, the POV value and the TBARs value are obviously reduced, meanwhile, no nitrite residue is left, and better sensory quality is achieved; the Linear Discriminant Analysis (LDA) of the electronic nose is shown in FIG. 6, which can effectively distinguish the differences of the three groups of preserved meat. The gas chromatography-mass spectrometry (GC-MS) analysis result shows that the relative contents of volatile substances related to fat oxidation, such as aldehydes, ketones, alcohols and the like, of the preserved meat after the yellow tea polysaccharide is added are 16.093%, 4.588% and 13.208% respectively, which are lower than those of blank groups (19.363%, 16.203% and 24.246%). The relative content of phenolic substances of flavor substances characteristic to the preserved meat of the yellow tea polysaccharide treated group is 61.361%, which is higher than that of the blank group (29.619%), which shows that the addition of the yellow tea polysaccharide in the preserved meat can better improve the physicochemical properties and flavor of the product and improve the quality of the preserved meat.
The result shows that the preserved meat can be well prepared by the process of adding tea polysaccharide without additionally adding nitrite, no nitrite is detected in the preserved meat, simultaneously, sensory indexes related to the preserved meat are not obviously different from those of a group with normal nitrite, and the relative contents of volatile substances such as aldehydes, ketones, alcohols and the like related to fat oxidation are 16.481%, 5.233% and 14.582% which are lower than those of a blank group (19.363%, 16.203% and 24.246%). The relative content of phenolic substances of flavor substances characteristic to the preserved meat of the yellow tea polysaccharide treatment group is 58.281%, which is higher than that of the blank group (29.619%), so that the physical and chemical properties and flavor of the preserved meat can be better improved by adding the yellow tea polysaccharide into the preserved meat without using nitrite, and the quality of the preserved meat is improved.
Therefore, the yellow tea polysaccharide prepared by the invention has excellent physicochemical properties due to the unique composition, and can fully utilize the antibacterial property, the oxidation resistance and the color protection property of the yellow tea polysaccharide in the improvement of the quality of the preserved meat, obviously reduce the nitrite in the preserved meat, enrich the flavor substance components in the preserved meat, and endow part of the preserved meat with the nutrition and health care functions of the yellow tea polysaccharide.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (6)
1. A yellow tea polysaccharide composition is characterized in that: the preparation method comprises the following steps:
1) preprocessing yellow tea leaves: cleaning yellow tea leaves, drying, crushing, sieving to prepare yellow tea dry powder, sealing, drying and storing;
2) weighing a proper amount of dry yellow tea powder, and performing ultrahigh pressure extraction through a proper amount of solvent;
3) centrifuging the extractive solution at 5000-7000r/min for 5-15min, and collecting supernatant;
4) concentrating the supernatant to 10% -20% of the original volume, adding 90% ethanol solution with 4-6 times of the volume of the concentrated solution, and precipitating for 24 h;
5) collecting precipitate, washing the precipitate with anhydrous ethanol and diethyl ether for 2-3 times, and dissolving the washed precipitate in water;
6) mixing the re-dissolved solution with Sevage solvent according to the volume ratio of 3:1, deproteinizing, collecting supernatant, and collecting for 2 times;
7) mixing the two supernatants, dialyzing the supernatant with 8kDa dialysis bag, and freeze-drying at-80 deg.C to obtain yellow tea polysaccharide composition powder;
the extraction solvent in the step 2) is distilled water, and the extraction time is 3-15 minutes; the extraction pressure is 200-; the material-liquid ratio is 1:5-25 g/mL;
the yield of the yellow tea polysaccharide composition is 9.32-10.81%, the yellow tea polysaccharide composition mainly comprises 26.1-28.9% of neutral sugar, 35.09-38.42% of uronic acid, 10-12.5% of polyphenol and 3.8-4.2% of protein, the neutral sugar consists of mannose, rhamnose, ribose, glucose, galactose, xylose, arabinose and fucose 8 sugars, and the molar percentage is 20.46-23.12%, 8.37-11.61%, 3.19-4.52%, 17.54-20.97%, 15.59-17.14%, 8.02-9.35%, 13.49-15.85% and 2.76-3.63%.
2. The yellow tea polysaccharide composition of claim 1, wherein: the tea is yellow tea or old yellow tea;
the yellow tea is dried by blowing after being cleaned, the drying temperature is 60-80 ℃, and the yellow tea is crushed and then sieved by a sieve of 90-100 meshes.
3. Yellow tea polysaccharide composition according to claim 1 or 2, characterized in that:
centrifuging the extractive solution at 6000r/min for 5 min;
concentrating the supernatant to 15% of the original volume;
adding 90% ethanol solution with 5 times volume of the concentrated solution, and precipitating for 24 h.
4. Yellow tea polysaccharide composition according to claim 1 or 2, characterized in that: the Sevage solvent in the step 6) is a mixed solution of chloroform and n-butanol with the volume ratio of 4: 1.
5. The use of the yellow tea polysaccharide composition as claimed in any one of claims 1 to 4 in cured meat, wherein the yellow tea polysaccharide composition is applied to the curing process of cured meat, and a vacuum tumbling curing method is adopted, so that the yellow tea polysaccharide composition can improve the quality of the cured meat, including remarkably reducing the content of nitrite and improving flavor substances in the cured meat.
6. The use of the yellow tea polysaccharide composition of claim 5 in bacon, wherein the bacon with good organoleptic properties can be obtained by pickling the yellow tea polysaccharide composition without adding nitrite in the process of bacon preparation.
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