CN113702134A - Sample preparation method of bamboo shoot meat analysis sample and application of sample - Google Patents
Sample preparation method of bamboo shoot meat analysis sample and application of sample Download PDFInfo
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- CN113702134A CN113702134A CN202111067098.9A CN202111067098A CN113702134A CN 113702134 A CN113702134 A CN 113702134A CN 202111067098 A CN202111067098 A CN 202111067098A CN 113702134 A CN113702134 A CN 113702134A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Preparation Of Fruits And Vegetables (AREA)
Abstract
The invention discloses a sample preparation method of bamboo shoot meat analysis samples, which takes bamboo shoots to be chemically and biologically analyzed as raw materials and comprises the following steps: selecting raw materials, cleaning, removing bases, punching and sampling, chopping, uniformly mixing, freezing, packaging and refrigerating to obtain the bamboo shoot meat analysis sample. The invention can obtain sample points with different aging degrees and different physiological activities, overcomes the defects of the existing sample preparation, solves the problems of uneven growth of bamboo shoots, poor representativeness of the traditional sampling method, large deviation of reliability of comparison among analysis batches and the like, can greatly improve the representativeness and the accuracy of the analysis of uneven bamboo shoot meat samples, reduce system errors, improve the reliability and the accuracy of analysis results, play a great role in the aspect of biological analysis of the bamboo shoots, and is particularly suitable for nutrient component analysis, heavy metal content analysis, pesticide residue analysis, antibiotic content analysis, post-harvest physiological and biochemical analysis and extraction, analysis and sampling of DNA or RNA of the bamboo shoots.
Description
The application is a divisional application with the application date of 2016, 11 and 25, the application number of 201611052523.6, and the name of the invention being 'a sample preparation method for bamboo shoot meat analysis samples'.
Technical Field
The invention belongs to the technical field of food analysis, and particularly relates to a sample preparation method of a bamboo shoot meat analysis sample for chemical and biological analysis of bamboo shoots.
Background
China is at the center of the distribution of bamboo in the world, and the area of the bamboo forest accounts for about 30 percent of the total area of the bamboo forest in the world. The edible part of bamboo is bamboo shoot, and Chinese bamboo shoot has been eaten for more than 3000 years. Bamboo shoots in bamboo species in China can be eaten by at least more than 200 kinds, and become one of the main forest vegetables in China. With the progress of human civilization, bamboo shoots have been developed from self-use to commercial products in the time of western Han dynasty. The medicinal value and the health care efficacy of bamboo shoots are widely recognized in China folks, and the traditional Chinese medicine considers that the bamboo shoots are sweet and cold in nature and taste, enter lung and stomach channels, have the effects of detoxifying and promoting eruption, clearing heat and reducing phlegm, and harmonizing the middle-jiao and moistening the intestines. In recent years, bamboo shoots are also found to have various health-care functions of losing weight, preventing intestinal cancer, reducing blood fat, resisting aging, enhancing immunity of organisms and the like, have a certain food therapy effect, and are novel health-care food. Currently, in japan, canada, korea, etc., many people consider bamboo shoots as one of the vegetables that must be eaten every day, which is indispensable for a meal. Bamboo shoot food is also popular with consumers in countries such as europe and america.
In order to discuss the relationship between the content of heavy metals in bamboo shoots and the heavy metals in soil, bamboo shoot samples in a main production area are collected for determination, and the average mass fractions of the heavy metals cadmium (Cd), lead (Pb), zinc (Zn), chromium (Cr) and copper (Cu) of the phyllostachys pracecox shoots are respectively 0009 mg-kg-1，0093mg·kg-1，8780mg·kg-1，0026mg·kg-1And 0830mg kg-1.5 Phyllostachys nigra (Carr.) Mitford cv. Pubescens) in Shaoxing city213 spring bamboo shoot samples are collected in the production area, the content of 5 heavy metals of arsenic, lead, cadmium, mercury and chromium is measured, and the detection rate of the 5 heavy metals exceeds 98 percent.
The annual yield of bamboo shoots in China is close to 1000 ten thousand tons, the yield of the fresh bamboo shoots is increased year by year, and the biological analysis of the bamboo shoots cannot be kept away from the fresh-keeping research of the fresh bamboo shoots.
No matter the analysis of the nutrition quality of the bamboo shoots, the analysis of heavy metals, the analysis of pesticide residues, the biological analysis of physiology, biochemistry and the like, the sampling and the sample preparation are not required, and the method is the most basic guarantee and the most key step for the accurate and reliable analysis. The previous sampling methods are all sample sampling methods with uniform texture, such as halving, quartering, diagonal dividing and the like, and are applied to irregular samples, so that the introduced system error is large; the main problems of the sampling method are as follows: firstly, because bamboo shoots are different from traditional vegetables and fruits, the bamboo shoots belong to buds in plant biological organ classification, and are large tender shoots, life activities are particularly vigorous, different areas on the whole buds are different, the whole bamboo shoot meat tissue of the bamboo shoots as the buds has no uniformity, the sampling is difficult to achieve representativeness, and the system error brought by sampling factors is large. Secondly, when the traditional sampling technology, namely a bisection or quartering method is adopted, ideal equipartition is difficult to achieve due to the irregular shape of the bamboo shoots, and the samples do not have the characteristics of equipartition. Thirdly, although the heterogeneity of the whole bamboo shoot can be overcome by using the whole bamboo shoot as a sample, the sample amount is large, on one hand, the storage is difficult, on the other hand, only a small part of the sample can be taken during specific analysis, and therefore, systematic error factors are not really eliminated. Fourth, especially in cell research and DNA & RNA extraction, the amount of sample required is very small, and conventional sampling methods cannot be widely representative.
With the development of scientific technology and the enhancement of health concept of people, the quality and safety of food are more emphasized, the pollution trend of heavy metal and pesticide is expanded, and the risk of the traditional sampling and sample preparation method is increased. The sample preparation method has important application value for analyzing the plant samples with heterogeneity of texture and irregular shape.
Disclosure of Invention
In view of the above problems in the prior art, an object of the present invention is to provide a method for preparing an analysis sample of bamboo shoot meat for chemical and biological analysis of bamboo shoots, which is particularly suitable for analysis of nutrient components, analysis of heavy metals, analysis of pesticide residues, analysis of antibiotic contents, post-harvest physiological and biochemical analysis, and extraction and analysis of DNA or RNA of bamboo shoots.
The invention provides a sample preparation method of an analysis sample of bamboo shoot meat, which comprises the following steps:
1) removing the base part of the bamboo shoots to obtain the bamboo shoots;
the thickness of the base part is 1-3 mm;
2) inserting a puncher into the bamboo shoot flesh tissue of the sample bamboo shoot from the bottom of the sample bamboo shoot along the direction parallel to the bamboo shoot shell of the sample bamboo shoot to obtain a bamboo shoot flesh sample strip;
the distribution of the bamboo shoot meat sample strips at the bottoms of the bamboo shoots comprises: 3, 5, 7 or 9 points;
3) crushing the bamboo shoot sample strips to obtain bamboo shoot particles;
the diameter of the bamboo shoot particles is less than or equal to 1 mm;
4) uniformly placing the bamboo shoot particles at the bottom of a container, and adding liquid nitrogen into the container for freezing to obtain a sample;
the volume ratio of the bamboo shoot particles to the container comprises 1 (3-5);
the method for adding the liquid nitrogen comprises batch injection, wherein the number of the batch injection comprises the following steps: a first implant, a second implant, and a third implant;
during the first injection and the second injection, the volume ratio of the liquid nitrogen to the broken bamboo shoot particles is (0.9-1.2): 1; when the third injection is carried out, the volume ratio of the liquid nitrogen to the volume of the bamboo shoot particles is (0.4-0.6): 1;
5) placing the sample in a freezing-resistant container for cold storage and standby;
the material of the freeze-resistant container comprises polyvinyl fluoride;
the volume ratio of the sample to the freeze-tolerant container comprises 1 (2-3);
the refrigerating temperature comprises-86 ℃ to-40 ℃.
Preferably, the bamboo shoot comprises a phyllostachys pracecox shoot, a phyllostachys glauca shoot or a phyllostachys pubescens shoot.
Preferably, the bamboo shoots comprise bamboo shoots which have no plant diseases, insect pests, erosion plaques, no mechanical injury and no microbial growth.
Preferably, when the sample is applied to biological analysis, the bamboo shoot includes fresh bamboo shoots with glossy bamboo shoot shells or fresh bamboo shoots without mildew and rot spots.
Preferably, the bamboo shoots include fresh bamboo shoots with glossy shells selected from a large number of bamboo shoot raw materials, or bamboo shoots which have undergone a preservation treatment for a different storage time and are kept away from mold and rot spots.
Preferably, the bamboo shoots are cleaned before the step 1);
the cleaning method comprises the following steps: removing silt on the bamboo shoot base and the bamboo shoot shell, and washing with clear water;
the cleaning time is less than or equal to 3-10 min;
the water temperature for rinsing with clean water is 10-30 ℃.
Preferably, the selected bamboo shoot further comprises a cleaning process, and the cleaning process comprises: washing the selected bamboo shoots in a water pool or basin, washing away silt on the surface of the base and the bamboo shoot shells, and washing with clean water.
Preferably, the operation of removing the base includes: the bamboo shoots are placed on a wood chopping block or a plastic chopping block, and the base is cut off by a stainless steel knife.
Preferably, the perforator in step 2) further comprises sterilization before use.
Preferably, the material of the puncher comprises a stainless steel puncher.
Preferably, the punch further comprises a pre-treatment before use; the pretreatment comprises the following steps: soaking the stainless steel puncher in a 1-4% sodium hydroxide dilute solution for 5-30 minutes, then cleaning the puncher with tap water until the cleaning solution is neutral, and finally washing the puncher with purified water or distilled water for 2-3 times; wrapping a puncher with packaging paper, putting the puncher into an electric heating air blowing drying oven, carrying out dry heat sterilization at 130-150 ℃ for 1.5-3 hours, and cooling to room temperature for later use;
preferably, in the step 2),
when the diameter of the cross section of the bottom of the bamboo shoot is 3-5 cm, the aperture of the puncher is 0.3-0.5 cm;
when the diameter of the cross section of the bottom of the bamboo shoot is 6-15 cm, the aperture of the puncher is 0.5-0.7 cm;
when the diameter of the cross section of the bottom of the bamboo shoot is larger than 15cm, the aperture of the puncher is 0.7-1.0 cm.
Preferably, the method of pulverizing comprises: the bamboo shoot meat sample strips are put on a clean and sanitary chopping block and cut up by a knife.
Preferably, the container comprises: an enamel cup or a stainless steel cup.
Preferably, the method for uniformly distributing the bamboo shoot particles comprises the following steps: shaking, shaking up and down or stirring with glass rod for several times.
Preferably, the volume ratio of the liquid nitrogen to the bamboo shoot particles in the first injection and the second injection is 1: 1; and when the third injection is carried out, the volume ratio of the liquid nitrogen to the bamboo shoot particles is 0.5: 1.
Preferably, when the liquid nitrogen is injected into the container in batches, the container is shaken or stirred by a stainless steel rod, so that the broken bamboo shoot grains are not bonded.
Preferably, when no gas overflow is observed at the container mouth and white bamboo shoot particles are obtained, the injection of liquid nitrogen is stopped.
The invention also provides the application of the sample preparation method in the preparation of samples of solid or plant tissues and organs with uneven texture.
The invention also provides application of the sample prepared by the sample preparation method in nutrient component analysis, heavy metal content analysis, pesticide residue analysis, antibiotic content analysis, post-harvest physiological and biochemical analysis, DNA extraction analysis or RNA extraction analysis of bamboo shoots.
By adopting the technology, compared with the prior art, the invention has the following beneficial effects:
1) according to the method, samples of the bamboo shoots are prepared by adopting the method, sample points with different aging degrees and different physiological activities can be obtained, the defects of the existing sample preparation are overcome, the problems of bamboo shoot growth heterogeneity, poor representativeness of the traditional sampling method, large deviation of reliability of comparison among analysis batches and the like are solved, the representativeness and the accuracy of heterogeneous bamboo shoot meat sample analysis can be greatly improved, the system error is reduced, the reliability and the accuracy of an analysis result are improved, and a great effect is played on the aspect of biological analysis of the bamboo shoots;
2) the invention provides a brand-new sample preparation method for bamboo shoot analysis samples, which is particularly suitable for analysis of nutrient components, analysis of heavy metal content, analysis of pesticide residues, analysis of antibiotic content, post-harvest physiological and biochemical analysis and extraction and analysis of DNA or RNA of bamboo shoots, can give a greater play to the representativeness of the analysis samples by the method, and provides a more scientific and reasonable basis for quality evaluation and quality control of bamboo shoot products; providing a sample in a real sense for the biological research of the picked bamboo shoots;
3) the method can control the sampling amount of the sample, but does not influence the representativeness of the sample, can take a large amount of samples, can also take a very small amount of samples, is particularly preferable for analyzing genes and the like, is flexible in sampling, and can realize the wide representativeness of the sample when a small amount of samples are taken for analyzing the samples;
4) the method provided by the invention has strong representativeness, is not only suitable for sampling all bamboo, but also suitable for preparing samples of solid samples with uneven texture or samples of plant tissues, organs and the like, provides an active thinking mode for analyzing the samples and eliminating system errors, and is suitable for popularization and application.
Drawings
FIG. 1a is a schematic view of a punching angle of a puncher for a phyllostachys pracecox shoot;
FIG. 1b is a schematic view of the punching angle of the moso bamboo shoot by the puncher;
FIG. 1c is a schematic view of the punching angle of the green bamboo shoot by the puncher;
FIG. 2 is a schematic cross-sectional view of a 3-point punch sampling;
FIG. 3 is a schematic cross-sectional view of a 5-point punch sampling;
FIG. 4 is a schematic cross-sectional view of a 7-point punch sampling;
FIG. 5 is a schematic cross-sectional view of a 9-point punch sampling.
In the figure: 1-a hole puncher.
Detailed Description
The invention is further described with reference to the following examples, but the scope of the invention is not limited thereto:
the invention discloses a sample preparation method of bamboo shoot meat analysis samples, which takes bamboo shoots to be analyzed as raw materials and is prepared by the steps of selecting, cleaning, removing bases, punching, sampling, chopping, uniformly mixing, freezing, packaging and refrigerating, wherein the sample preparation process comprises the following steps: selecting raw materials → cleaning → removing base portion → sampling by punching → cutting up → mixing → freezing → packaging → refrigerating → sampling, the concrete steps are as follows:
1) selecting raw materials from a large amount of bamboo shoots to be analyzed, selecting the bamboo shoots which have no plant diseases, insect pests, erosion spots, mechanical injuries and microorganism growth, and during biological analysis, requiring fresh bamboo shoots with glossy bamboo shoot shells or bamboo shoots which have been subjected to other fresh-keeping treatments and have different storage times to avoid mildew and rot spots;
2) cleaning, namely cleaning the selected bamboo shoots in a water pool or a water basin to remove silt on the surfaces of the base parts and the bamboo shoot shells; then washing with clean water, wherein the temperature of the water for washing is controlled to be 10-30 ℃, and the whole washing time is controlled within 3-10 minutes;
3) removing the basal part, putting the cleaned bamboo shoots on a wood chopping board or a plastic chopping board, and cutting off the basal part by a stainless steel knife to be 1-3 mm thick;
4) preparing a puncher, soaking the stainless steel puncher in dilute sodium hydroxide alkali liquor for 5-30 minutes for punching and sampling, cleaning the stainless steel puncher with tap water until the cleaning solution is nearly neutral, and finally washing the stainless steel puncher with purified water or distilled water for 2-3 times; wrapping a hole puncher with packaging paper, putting the wrapped hole puncher into an electrothermal blowing drying oven, carrying out dry heat sterilization at 130-150 ℃ for 1.5-3 hours, cooling to room temperature for later use, inserting the hole puncher into bamboo shoot meat tissues from the base of the bamboo shoots along the direction parallel to the bamboo shoot shells of the base, and distributing sampling insertion points at the base according to the cross section of the base of the bamboo shoots in a 3-point mode, a 5-point mode, a 7-point mode or a 9-point mode, wherein as shown in figures 2-5, as shown in figure 2, the 3-point mode has two punching modes, and three holes are arranged on a straight line or in a triangle.
The method comprises the steps of customizing a puncher 1 according to the length of bamboo shoots, wherein the length of the puncher is close to the length of bamboo shoot meat of the bamboo shoots; the aperture of the puncher 1 is determined according to the thickness of the bamboo shoots, the puncher with smaller selective aperture is used for selecting the thinner bamboo shoot body of the bamboo shoots, and the puncher with larger selective aperture is used for selecting the thicker bamboo shoot body. As shown in fig. 1a, 1b and 1c, the bamboo shoot body of the phyllostachys pracecox shoot is elongated, and a punch 1 having a small aperture is selected; the moso bamboo shoots are thick and straight, and a puncher 1 with large aperture is selected to punch holes in the straight direction; roughly bending the body of the green bamboo shoot, selecting a puncher 1 with a large aperture, and obliquely punching; generally, the length of the punch 1 is 10 to 30 cm, and the diameter of the hole of the punch 1 is 0.3 to 1 cm. Selecting bamboo shoots with the cross section of the bamboo shoot base part of 3-5 cm in diameter and a puncher with the hole diameter of 0.3-0.5 cm; selecting a perforator with the diameter of 0.5-0.7 cm for bamboo shoots with the diameter of 6-15 cm at the cross section of the bamboo shoot base; selecting bamboo shoots with the diameter of more than 15cm at the cross section of the bamboo shoot base part, and selecting a puncher with the aperture of 0.7-1 cm; sampling quantity is large, and a puncher with a large aperture is selected; sampling quantity is small, and a puncher with small aperture is selected;
5) cutting bamboo shoot sample taken out by the puncher on a clean and sanitary chopping board, and chopping the bamboo shoot sample with a knife, wherein the diameter of bamboo shoot particles is not more than 1 mm;
6) uniformly mixing, namely putting bamboo shoot particles into a container such as an enamel cup or a stainless steel cup, wherein the bamboo shoot amount accounts for 1/5-1/3 of the volume of the container, and fully shaking, shaking up and down or stirring for a plurality of times by using a glass rod;
7) freezing, injecting liquid nitrogen with the volume close to that of the broken bamboo shoots into a container containing the uniformly mixed broken bamboo shoots, shaking the container or stirring by using a stainless steel bar, and injecting the liquid nitrogen with the same volume again and stirring when the opening of the container cannot see the gas overflow so as to ensure that the broken bamboo shoots are not bonded. And injecting liquid nitrogen with the volume of about 1/2 of the bamboo shoot particles for the third time, and fully shaking the container until no gas overflows from the opening of the container. At the moment, the bamboo shoot particles are white;
8) packaging bottles with proper sizes are selected according to the amount of samples, frozen bamboo shoot particles are poured into an anti-freezing polyvinyl fluoride bottle in time, the filling amount is 1/2-2/3 of the volume of the bottle, a cover is covered, and a label is pasted;
9) and (4) freezing and storing the packaged sample immediately in an ultralow temperature refrigerator at-86 to-40 ℃, and taking the sample according to the required amount during analysis.
Example 1
1. Selecting fresh bamboo shoots which have no plant diseases, insect pests and erosion spots on bamboo shoots and have no mechanical injury on bamboo shoot bodies from commercially available bamboo shoots;
2. cleaning raw materials, namely cleaning selected bamboo shoots in a water tank, and washing away silt carried on the surfaces of bases and bamboo shoot shells by using tap water at the temperature of 10-20 ℃; then the water is used for washing. Cleaning within 3-10 minutes;
3. removing the base part, placing the cleaned bamboo shoots on a clean chopping board, and cutting off the base part by a stainless steel knife to be 3mm thick;
4. punching and sampling, namely longitudinally inserting bamboo shoot tissues from the base part of each bamboo shoot by using a small-size puncher which is cleaned and sterilized, sampling 3 points of the base part of each bamboo shoot, wherein the insertion points are distributed on a straight line, one central point is arranged, and the other two points are taken at the middle position of the wall thickness;
5. cutting the bamboo shoot and meat strips on a clean and sanitary chopping board into particles with the diameter not more than 1 mm;
6. uniformly mixing, putting the bamboo shoot particles into a stainless steel basin, fully shaking the bamboo shoot particles until the thickness of the bamboo shoot particles is about 1/3 of the depth of the stainless steel basin, and stirring the bamboo shoot particles for a plurality of times by using a glass rod until the bamboo shoot particles are uniform;
7. freezing, injecting liquid nitrogen with the volume close to that of the bamboo shoot particles into a container containing the uniformly mixed bamboo shoot particles, stirring by using a stainless steel rod, injecting liquid nitrogen with the same volume again and stirring when gas cannot be seen, so that the particles are not bonded, injecting liquid nitrogen with the volume about 1/2 of the bamboo shoot particles for the third time, and fully shaking until the gas cannot be seen to overflow;
8. packaging, namely timely transferring the frozen bamboo shoot particles into a polyvinyl fluoride bottle, filling 1/2-2/3 of the volume of the bottle, covering the bottle with a cover, and sticking a label;
9. freezing and storing the packaged sample immediately in an ultralow temperature refrigerator at-80 ℃ for multiple times.
Example 2
The method is used for preparing a moso bamboo shoot sample for analyzing the heavy metal content and the pesticide residue, and comprises the following steps:
1. selecting raw materials, digging bamboo shoots in a moso bamboo shoot producing area or purchasing the bamboo shoots in a farmer market, then selecting the bamboo shoots without plant diseases and insect pests and erosion spots on bamboo shoots bodies, and taking the bamboo shoots without mechanical injury except for cuts as the raw materials for sampling;
2. cleaning the selected raw materials are placed in a cleaning basin, and tap water at room temperature (about 20 ℃) is used for cleaning mud on the base and the bamboo shoot shell surface; and then washed with distilled water. The cleaning time is controlled within 5-8 minutes;
3. removing the basal part, placing the cleaned bamboo shoot on a wood chopping board, and cutting off the basal part by a stainless steel knife to be 2.5mm thick;
4. punching and sampling as shown in FIG. 1b, inserting bamboo shoot flesh tissue from the base of moso bamboo shoot along the direction parallel to the bamboo shoot shell of the base by using a cleaning and sterilizing puncher, wherein the insertion points are 1 central point and the rest 8 points are approximately and uniformly distributed near the circumference of 1cm away from the edge as shown in FIG. 5;
5. cutting, namely putting the bamboo shoot meat sample strips taken out in the steps on a clean and sanitary chopping board, and cutting into particles with the diameter of 0.8 mm;
6. uniformly mixing, namely putting bamboo shoot particles into a stainless steel cup, wherein the bamboo shoot amount accounts for 1/4-1/3 of the volume of the container, and stirring for a plurality of times by using a glass rod;
7. freezing, injecting liquid nitrogen with the volume close to that of the bamboo shoot particles into a stainless steel cup containing the uniformly mixed bamboo shoot particles, shaking until no gas overflows from the cup mouth, injecting liquid nitrogen with the same volume and stirring again to ensure that the particles are not bonded, injecting liquid nitrogen with the volume about that of the bamboo shoot particles 1/2 for the third time, and fully shaking until no gas overflows;
8. packaging, pouring the frozen bamboo shoot particles into a 1000ml polyethylene bottle in time, filling 2/3 bottles with the volume being equal to the volume of the bottle, covering the bottle with a cover, and sticking a label;
9. freezing and storing the packaged sample in an ultralow temperature refrigerator at-40 ℃ immediately, and taking the sample according to the required amount during analysis.
Example 3
The method comprises the following specific steps of preparing a physiological and biochemical index analysis sample of bamboo shoot meat of green bamboo shoots:
1. the selection of the bamboo shoot raw materials is to select bamboo shoots which have no plant diseases, insect pests and erosion spots on the bamboo shoot bodies, no mechanical injury except the cuts and glossy bamboo shoot shells from the newly picked bamboo shoots in a green bamboo forest land of Pingyangcun Mazhou province from 7-9 months. Selecting bamboo shoots which are stored for a certain time interval and have no mildew and rot spots as sampling raw material bamboo shoots in the later fresh-keeping process;
2. washing the selected initial raw materials by tap water to remove mud on the surfaces of the base part and the bamboo shoot shell; then washing with pure water of Waha. The temperature of the water for cleaning is controlled between 20 ℃ and 30 ℃, and the whole cleaning time is controlled within 10 minutes. The fresh-keeping bamboo shoots at the later stage do not need to be cleaned;
3. removing a basal part, putting the cleaned bamboo shoots on a plastic chopping board, and cutting off the basal part by a stainless steel knife to be 1-2 mm thick;
4. punching and sampling, as shown in figure 1c, inserting bamboo shoot flesh tissues from the center of the base of the bamboo shoot and a point which is 0.5-1 cm away from the edge on a crossed cross line of the cross by using a puncher which is cleaned and sterilized in advance along the direction parallel to the bamboo shoot shell of the base, and sampling 5 points of each bamboo shoot as shown in figure 3;
5. chopping, placing the bamboo shoot stick on clean and sanitary chopping board, and chopping with knife. The diameter of the bamboo shoot particles is 0.6 mm;
6. uniformly mixing, putting bamboo shoot particles into a 1000ml enamel cup, wherein the bamboo shoot amount accounts for 1/5 of the volume of the container, and fully shaking, shaking and vertically shaking for a plurality of times;
7. freezing, injecting about 200ml of liquid nitrogen into an enamel cup containing uniformly mixed bamboo shoot particles, shaking until no gas appears at the cup mouth, injecting the liquid nitrogen with the same volume again and stirring to ensure that the particles are not bonded, injecting the liquid nitrogen with the volume of about 100ml for the third time, and fully shaking the enamel cup until no gas appears. At the moment, the bamboo shoot particles are white;
8. packaging, pouring the frozen bamboo shoot particles into a low-temperature-resistant polyvinyl fluoride bottle with the volume of 200ml in time, filling 2/3 of the volume of the bottle, covering the bottle with a cover, and labeling;
9. freezing and storing the packaged sample in an ultra-low temperature refrigerator at-86 ℃.
In conclusion, the sample preparation method provided by the invention is simple to operate and convenient to popularize, and is suitable for bamboo shoots of various types and shapes.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (9)
1. A sample preparation method of an analysis sample of bamboo shoot meat comprises the following steps:
1) removing the base part of the bamboo shoots to obtain the bamboo shoots;
the thickness of the base part is 1-3 mm;
2) inserting a puncher into the bamboo shoot flesh tissue of the sample bamboo shoot from the bottom of the sample bamboo shoot along the direction parallel to the bamboo shoot shell of the sample bamboo shoot to obtain a bamboo shoot flesh sample strip;
the distribution of the bamboo shoot meat sample strips at the bottoms of the bamboo shoots comprises: 3, 5, 7 or 9 points;
3) crushing the bamboo shoot sample strips to obtain bamboo shoot particles;
the diameter of the bamboo shoot particles is less than or equal to 1 mm;
4) uniformly placing the bamboo shoot particles at the bottom of a container, and adding liquid nitrogen into the container for freezing to obtain a sample;
the volume ratio of the bamboo shoot particles to the container comprises 1 (3-5);
the method for adding the liquid nitrogen comprises batch injection, wherein the number of the batch injection comprises the following steps: a first implant, a second implant, and a third implant;
during the first injection and the second injection, the volume ratio of the liquid nitrogen to the broken bamboo shoot particles is (0.9-1.2): 1; when the third injection is carried out, the volume ratio of the liquid nitrogen to the volume of the bamboo shoot particles is (0.4-0.6): 1;
5) placing the sample in a freezing-resistant container for cold storage and standby;
the material of the freeze-resistant container comprises polyvinyl fluoride;
the volume ratio of the sample to the freeze-tolerant container comprises 1 (2-3);
the refrigerating temperature comprises-86 ℃ to-40 ℃.
2. A method as claimed in claim 1, wherein the bamboo shoots comprise phyllostachys pracecox shoots, phyllostachys glauca shoots or phyllostachys pracecox shoots.
3. A sample preparation method as claimed in claim 1, wherein said bamboo shoots comprise bamboo shoots having no plant or insect pest plaque, no mechanical injury and no microbial growth.
4. A method as claimed in claim 3, wherein the bamboo shoots, when the sample is subjected to biological analysis, comprise fresh bamboo shoots having a glossy shoot shell or fresh bamboo shoots having no mildew stains.
5. The sample preparation method according to claim 1, wherein the bamboo shoots are cleaned before the step 1);
the cleaning method comprises the following steps: removing silt on the bamboo shoot base and the bamboo shoot shell, and washing with clear water;
the cleaning time is less than or equal to 3-10 min;
the water temperature for rinsing with clean water is 10-30 ℃.
6. The method of claim 1, wherein the punch of step 2) further comprises sterilizing prior to use.
7. The sample preparation method according to claim 1, wherein in the step 2),
when the diameter of the cross section of the bottom of the bamboo shoot is 3-5 cm, the aperture of the puncher is 0.3-0.5 cm;
when the diameter of the cross section of the bottom of the bamboo shoot is 6-15 cm, the aperture of the puncher is 0.5-0.7 cm;
when the diameter of the cross section of the bottom of the bamboo shoot is larger than 15cm, the aperture of the puncher is 0.7-1.0 cm.
8. Use of the sampling method of any one of claims 1 to 7 for sampling of solid or plant tissue organs with inhomogeneous texture.
9. Use of the sample prepared by the sample preparation method according to any one of claims 1 to 7 in analysis of nutrient components, analysis of heavy metal content, analysis of pesticide residues, analysis of antibiotic content, post-harvest physiological and biochemical analysis, analysis of DNA extraction, or analysis of RNA extraction of bamboo shoots.
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| JP4368414B1 (en) * | 2009-04-20 | 2009-11-18 | 薫美 藤井 | Processed koji suitable for frozen storage, method for producing the same, and method for freezing koji |
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| CN103487397B (en) * | 2013-09-23 | 2015-10-28 | 浙江农林大学 | A kind of thunder bamboo shoots hardness method for quick and device |
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| JP4368414B1 (en) * | 2009-04-20 | 2009-11-18 | 薫美 藤井 | Processed koji suitable for frozen storage, method for producing the same, and method for freezing koji |
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