CN112180006A - Method for constructing tracing of apostichopus japonicus origin - Google Patents
Method for constructing tracing of apostichopus japonicus origin Download PDFInfo
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- G01N30/02—Column chromatography
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Abstract
The invention discloses a method for constructing a traceability of a production place of stichopus japonicus, which comprises the following steps: s1, obtaining the optimal feeding concentration range of various trace elements of the stichopus japonicus through a test for pre-judging the optimal feeding concentration range of the trace elements of the stichopus japonicus at a specific growth and development stage, and then preparing a standard stichopus japonicus feed; s2, adopting standard stichopus japonicus feed to carry out feeding test on stichopus japonicus at a specific growth and development stage, detecting the attenuation and deposition rules of trace elements in the stichopus japonicus, and constructing a standard fingerprint of the deposition amount of various trace elements; s3, detecting the content of trace elements in the stichopus japonicus to be detected, comparing the content with a standard fingerprint, and tracing the origin, quality and authenticity of the stichopus japonicus to be detected. The invention realizes the identification of the identity, quality and producing area of the stichopus japonicus by adding the trace elements which are beneficial to the growth and development of the stichopus japonicus into the stichopus japonicus feed, performing feeding tests on the stichopus japonicus and constructing the standard fingerprint spectrum by utilizing the deposition rule of the trace elements in the stichopus japonicus.
Description
Technical Field
The invention relates to the technical field of analysis and test, in particular to a construction method for tracing the origin of stichopus japonicus.
Background
The stichopus japonicus is mainly produced in an intertidal zone on the coast of yellow Bohai sea, is one of main culture economic types in the coastal region of the northern China, and has unique flavor and higher medical and health care effects. The body wall of the stichopus japonicus is the main edible part, is rich in various nutrient substances and trace elements, and is popular with many consumers.
However, the stichopus japonicus breeding industry is developed rapidly at present, the quality of the stichopus japonicus feed is uneven, and the healthy development of the stichopus japonicus breeding industry is restricted, so that the quality of the stichopus japonicus commodity is uneven, and the phenomenon of sub-quality is caused.
Therefore, the invention not only can promote the growth and development of the stichopus japonicus in different stages and enhance the immunity of the organism of the stichopus japonicus, but also can help the stichopus japonicus to safely live in summer (when the water temperature exceeds 25 ℃, the stichopus japonicus enters a sleep-aiding state, the stichopus japonicus at the stage can have the phenomena of metabolism weakening, body mass reduction, internal organ degeneration and the like), and a standard stichopus japonicus fingerprint map (the fingerprint map refers to DNA of certain complex substances, such as traditional Chinese medicines, certain organisms or certain tissues or cells, and after protein is properly treated, a chromatogram map or a graph capable of marking the chemical characteristics is obtained by adopting a certain analysis means). The existing fingerprints are mainly divided into traditional Chinese medicine fingerprints, DNA fingerprints and peptide fingerprints. The method can draw a standard fingerprint spectrum with excellent confidentiality by manually controlling the deposition amount of trace elements of each part (body wall, intestinal tract, bone piece and other tissues) of the stichopus japonicus according to a traditional Chinese medicine fingerprint spectrum technology, thereby not only standardizing the stichopus japonicus breeding industry, but also realizing the rapid tracing of the origin of the stichopus japonicus production place so as to conveniently identify the identity, the quality and the authenticity of the stichopus japonicus.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a method for constructing a stichopus japonicus origin tracing area.
In order to achieve the above object, the present invention provides a method for tracing the origin of stichopus japonicus, comprising the following steps:
s1, preparation of standard stichopus japonicus feed: the method comprises the steps of obtaining the optimal feeding concentration range of at least three trace elements of stichopus japonicus at a corresponding stage by carrying out a test for pre-judging the optimal feeding concentration range of the trace elements on the stichopus japonicus at a specific growth and development stage, and preparing a standard stichopus japonicus feed containing at least three trace elements in the optimal feeding concentration range according to the optimal feeding concentration range, wherein the optimal feeding concentration represents the concentration of the trace elements harmless to the healthy growth of the stichopus japonicus;
s2, constructing a standard fingerprint of the deposition amount of at least three trace elements in each part of the stichopus japonicus: carrying out a feeding test on the stichopus japonicus at a specific growth and development stage for a certain time by adopting the prepared standard stichopus japonicus feed, then detecting the attenuation condition and the deposition rule of the trace elements in each part of the stichopus japonicus body, and constructing a standard fingerprint of the deposition amount of at least three trace elements in each part of the stichopus japonicus body;
s3, tracing the source and the origin of the stichopus japonicus to be detected: and (4) tracing the origin, quality and authenticity of the stichopus japonicus to be detected by detecting the content of the trace elements of each part in the stichopus japonicus to be detected and comparing the content with the standard fingerprint obtained in the step S2.
Further, the trace elements comprise fat-soluble vitamins, water-soluble vitamins and mineral elements, wherein the fat-soluble vitamins comprise at least one of vitamin A, vitamin D and vitamin E, and the water-soluble vitamins comprise at least one of vitamin C, vitamin B1 and vitamin B2; the mineral elements include but are not limited to selenium, zinc, copper, cobalt, manganese, iron, calcium, magnesium; the specific growth and development stage is a seedling stage of the stichopus japonicus, a growth stage of young stichopus japonicus or a pre-summer stage of adult stichopus japonicus; and each part in the detected stichopus japonicus is the body wall, the intestinal tract or the bone fragment of the stichopus japonicus.
Further, the preparation of the standard stichopus japonicus feed comprises the following specific steps:
(1) crushing, sieving and gradually and fully mixing all feed raw materials for later use;
(2) putting the uniformly mixed feed raw materials into a fermentation tank, adding beneficial microorganisms, and fermenting the feed raw materials by utilizing a beneficial microorganism fermentation enzymolysis technology to obtain fermented feed;
(3) adding at least three trace elements which are in the optimal feeding concentration range into the fermented feed, fully stirring and uniformly mixing, and adding water to ensure that the water content is kept to be 20-40% of the total weight of the feed to obtain a prefabricated feed;
(4) and (3) placing the prefabricated material in an extrusion granulator for extrusion forming to obtain the standard stichopus japonicus feed.
Further, the adding mode of the at least three trace elements is a free combination mode in different combinations or different proportions within the optimal feeding concentration range; the addition mode of the trace elements is direct addition or microorganism enrichment.
Further, the beneficial microorganism is one or the combination of more than two of yeast, probiotic bacillus, clostridium butyricum, lactobacillus, bifidobacterium or actinomycetes, and the adding amount of the beneficial microorganism is 0.2-1% of the total weight of the feed.
Further, the fermentation condition is that the fermentation is carried out for 20-28h at room temperature.
The second purpose of the present invention is to provide the application of the construction method for tracing the origin of stichopus japonicus, which is used for identifying the origin, quality and authenticity of the origin of stichopus japonicus.
In the invention, the stichopus japonicus at a specific growth and development stage (stichopus japonicus offspring stage, young stichopus japonicus growth stage or adult stichopus japonicus pre-summer stage) is selected to be fed for a test, mainly because the stichopus japonicus at different growth and development stages have different attenuation and deposition rules on trace elements, for example, at the stichopus japonicus offspring stage and the young stichopus japonicus growth stage, the addition of suitable trace elements can promote growth and enhance the immunity of the organism; in the pre-summer stage of the adult stichopus japonicus, the adult stichopus japonicus can enter a sleep-in state when the water temperature exceeds 25 ℃, the metabolism of the adult stichopus japonicus is weakened, the body mass is reduced, the internal organs are degraded and the like, and sufficient energy supply is provided for the adult stichopus japonicus in the pre-summer stage, for example, a proper amount of trace element selenium and vitamin E are added, so that the immunity of the organism of the adult stichopus japonicus can be enhanced while the growth and development of the adult stichopus japonicus are promoted, the adverse reaction of the adult stichopus japonicus caused by high temperature is relieved, the adult stichopus japonicus can be safely exposed to summer, the stress injury can be quickly recovered, the environmental change is.
In the invention, the part of the stichopus japonicus to be detected is the body wall, intestinal tract or bone slice of the stichopus japonicus, and mainly different trace elements are deposited in different parts of the stichopus japonicus, such as high fat content in the digestive tract and easy enrichment of fat-soluble vitamins, such as vitamin A, vitamin D and vitamin E; most mineral elements and water-soluble vitamins such as selenium, zinc, copper, cobalt, manganese, iron, magnesium, vitamin C and the like are easily enriched in the body wall of the stichopus japonicus, and the corresponding content can be obtained by detecting the body wall of the stichopus japonicus; the mineral element calcium is more easily enriched in bone slices of the stichopus japonicus, and the content of the mineral element calcium can be obtained by detecting the bone slices in the body wall of the stichopus japonicus (wherein the bone slices in the body wall are obtained by putting the body wall into a test tube of 2mol/L sodium hydroxide, heating the body wall by an alcohol lamp until the body wall is dissolved, sucking out the sediment at the bottom of the test tube to be detected); the nutrient substances absorbed by the stichopus japonicus not only can be supplied for the stichopus japonicus to grow, but also can be enriched in each part in the stichopus japonicus, so that the attenuation rule of the trace elements can be obtained, and further, various trace elements in each part in the stichopus japonicus can be detected according to the attenuation rule.
Compared with the prior art, the invention has the following beneficial effects:
aiming at the physiological and metabolic characteristics of the stichopus japonicus, the raw materials of the stichopus japonicus feed are pretreated by mixed microbial fermentation and enzymolysis technology, so that macromolecular nutrient substances such as saccharides, lipids, proteins and the like in the raw materials of the feed are decomposed into micromolecular substances such as monosaccharide, polysaccharide, fatty acid, amino acid, polypeptide and the like which are beneficial to the digestion and absorption of the stichopus japonicus, the absorption and utilization rate of the feed is improved, the waste is avoided, trace elements (such as vitamin A, vitamin D, vitamin E, vitamin C, selenium, zinc, copper, cobalt, manganese, iron, calcium or magnesium) which are beneficial to the growth and development of the stichopus japonicus are added into the stichopus japonicus feed, and the feed is used for feeding the stichopus japonicus in specific production and development stages (offspring seeds, young stichopus japonicus growth stages and adult stichopus japonicus early-sleep stages) for a period of time, so that the growth and development of the stichopus japonicus are further promoted, and meanwhile, different trace elements, Bone fragments and body wall), and establishes standard fingerprint spectra with strong confidentiality for deposition amount of different microelements in each part of stichopus japonicus, thereby realizing identification of identity, quality and origin of stichopus japonicus.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples.
Example 1
The jew-loving sea cucumber is usually jew-loving sea cucumber which is really reputed as "crown of sea cucumber", and in this embodiment, the stichopus japonicus preliminarily judged to be jew-loving sea cucumber in appearance is taken as an example to identify whether the stichopus japonicus to be detected is jew-loving sea cucumber produced by this company. Firstly, carrying out a pre-determination optimal feeding concentration test on 2-year stichopus japonicus (weight: 140-; the following is described as a representative of the prejudged test to obtain the optimum range of feeding concentration of elemental selenium:
s1: preparation of standard stichopus japonicus feed: pulverizing basic materials (sea mud, yeast tablet, herba Zosterae Marinae, rainbow powder, scallop edge and Spirulina etc.), sieving with 100 mesh sieve, adding at a certain proportion, and mixing step by step; putting the uniformly mixed basic feed into a fermentation tank, adding a beneficial microorganism mixture (comprising saccharomycetes, probiotic spores, clostridium butyricum, lactobacillus, bifidobacteria and actinomycetes) in proportion, and fermenting the feed raw materials for 26 hours at room temperature by utilizing a beneficial microorganism fermentation enzymolysis technology to obtain a fermented feed; adding different gradient trace element selenium, taking a group without the selenium element as a control group, and carrying out a pre-determination test on the stichopus japonicus (in early summer sleeping period), wherein the experimental result shows that the feed with the feeding concentration of 0.5-1 mg/kg of the selenium element can obviously promote the growth of the stichopus japonicus (in early summer sleeping period) and ensure that the content of the selenium in the body wall of the stichopus japonicus is 0.45-0.56 mg/kg through selenium enrichment, so that the selenium-enriched stichopus japonicus standard is reached, and the optimal feeding concentration range of the selenium element is 0.45-0.56 mg/kg;
similarly, the optimal feeding concentration ranges of various trace elements (copper 4-7 mg/kg, zinc 17-30 mg/kg, cobalt 12-19 mg/kg, manganese 10-20 mg/kg, magnesium 300-400 mg/kg, iron 25-30 mg/kg and calcium 15-20 mg/kg) are respectively obtained by carrying out a pre-determination test on other trace elements;
according to the obtained optimal feeding concentration range of the multiple trace elements, freely selecting and adding four trace elements of selenium, copper, magnesium and manganese (wherein the concentration of selenium is 0.5 mg/kg, the concentration of copper is 5mg/kg, the concentration of magnesium is 300 mg/kg and the concentration of manganese is 20 mg/kg) into the fermented feed, fully stirring and uniformly mixing, adding water to keep the water content at 30% of the total weight of the feed to obtain a prefabricated material, and placing the prefabricated material into an extrusion granulator for extrusion forming to obtain the standard stichopus japonicus feed.
S2, constructing a standard fingerprint of the deposition rule of four trace elements including selenium, copper, magnesium and manganese of 2-year-old oplopanax elatus nakai: feeding test of 2-year-old stichopus japonicus (weight: 140-;
s3, tracing the source and the origin of the stichopus japonicus to be detected: detecting the contents of four trace elements of selenium, copper, magnesium and manganese in the body wall of the 2-year stichopus japonicus to be detected to obtain that the concentration of selenium in the body wall of the 2-year stichopus japonicus to be detected is 0.45 mg/kg, the concentration of copper is 1.55 mg/kg, the concentration of magnesium is 12.05mg/kg and the concentration of manganese is 23.2 mg/kg, comparing with the standard fingerprint spectrum to obtain that the four trace elements of selenium, copper, magnesium and manganese are all in the standard fingerprint spectrum, and thus judging that the stichopus japonicus to be detected is high-quality Liaoshen produced by Dalianxin Yulong ocean biology science and technology Limited.
Example 2
The method comprises the following steps of renting the young sea cucumber seeds produced by the Dalianxin Yulong ocean biology science and technology GmbH in a leasing mode, buying the young sea cucumber seeds after culturing for a period of time, feeding standard stichopus japonicus feed with strong confidentiality at the young sea cucumber stage, constructing standard fingerprint spectrums of various trace elements in the young sea cucumber bodies according to the deposition change rule of the trace elements in the young sea cucumber bodies (step S2 in reference example 1), and judging the sources and the qualities of the trace elements by detecting the content of the trace elements of all parts (intestinal tracts, bone fragments and body walls) in the purchased stichopus japonicus bodies and comparing the content with the standard fingerprint spectrums (step S3 in reference example 1), wherein the concrete implementation steps are as follows:
s1: preparation of standard stichopus japonicus feed: through the test of pre-judging the optimal feeding concentration range of the trace elements for the stichopus japonicus offspring seeds, the optimal feeding concentration ranges of at least three trace elements (vitamin C500-1500 mg/kg, selenium 0.5-1.5 mg/kg, zinc 17-30 mg/kg, cobalt 12-19 mg/kg, manganese 10-20 mg/kg, magnesium 300-400 mg/kg, calcium 15-20 mg/kg and the like) of the stichopus japonicus at the stage are obtained, preparing standard stichopus japonicus feed containing four trace elements of vitamin C, selenium, magnesium and zinc according to the obtained optimal feeding concentration range of the trace elements of the stichopus japonicus offspring seeds (wherein the concentration of the vitamin C is 500 mg/kg, the concentration of the selenium is 1 mg/kg, the concentration of the magnesium is 350 mg/kg and the concentration of the zinc is 30 mg/kg);
s2: constructing a standard fingerprint spectrum of the four trace element deposition rules of the stichopus japonicus in the seedling stage: the standard stichopus japonicus feed is adopted to regularly feed stichopus japonicus seedlings to obtain the standard fingerprint of the deposition amount of the four trace elements in the stichopus japonicus body wall (the content of the trace elements in the body wall is 200 mg/kg of vitamin C170-.
S3, tracing the source and the origin of the stichopus japonicus to be detected: detecting the content of the four trace elements in the body wall of the stichopus japonicus offspring seed to be detected to obtain the concentration of the vitamin C of 192mg/kg, the concentration of the selenium of 0.3 mg/kg, the concentration of the magnesium of 11.85mg/kg and the concentration of the zinc of 85mg/kg in the body wall, comparing the concentration with the standard fingerprint spectrum to obtain the content of the four trace elements of the vitamin C, the selenium, the magnesium and the zinc of all in the standard fingerprint spectrum, and judging the stichopus japonicus offspring seed to be detected to be the Liaoshen offspring seed produced by Dalianxin Yulong ocean biology science and technology Co.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (7)
1. A method for constructing a traceability of a production area of stichopus japonicus is characterized by comprising the following steps:
s1, preparation of standard stichopus japonicus feed: the optimal feeding concentration range of at least three trace elements of the stichopus japonicus at a corresponding stage is obtained by carrying out a test for pre-judging the optimal feeding concentration range of the trace elements on the stichopus japonicus at a specific growth and development stage, and a standard stichopus japonicus feed containing at least three trace elements in the optimal feeding concentration range is prepared according to the optimal feeding concentration range;
s2, constructing a standard fingerprint of the deposition amount of at least three trace elements in each part of the stichopus japonicus: carrying out a feeding test on the stichopus japonicus at a specific growth and development stage for a certain time by adopting the prepared standard stichopus japonicus feed, then detecting the attenuation condition and the deposition rule of the trace elements in each part of the stichopus japonicus body, and constructing a standard fingerprint of the deposition amount of at least three trace elements in each part of the stichopus japonicus body;
s3, tracing the source and the origin of the stichopus japonicus to be detected: and (4) tracing the origin, quality and authenticity of the stichopus japonicus to be detected by detecting the content of the trace elements of each part in the stichopus japonicus to be detected and comparing the content with the standard fingerprint obtained in the step S2.
2. The method as claimed in claim 1, wherein the trace elements include fat-soluble vitamins, water-soluble vitamins, and mineral elements, the fat-soluble vitamins include at least one of vitamin A, vitamin D, and vitamin E, and the water-soluble vitamins include at least one of vitamin C, vitamin B1, and vitamin B2; the mineral elements comprise at least one of selenium, zinc, copper, cobalt, manganese, iron, calcium and magnesium; the specific growth and development stage is a seedling stage of the stichopus japonicus, a growth stage of young stichopus japonicus or a pre-summer stage of adult stichopus japonicus; and each part in the detected stichopus japonicus is the body wall, the intestinal tract or the bone fragment of the stichopus japonicus.
3. The method for constructing the stichopus japonicus origin tracing of claim 1, wherein the preparation of the standard stichopus japonicus feed comprises the following specific steps:
(1) crushing, sieving and gradually and fully mixing all feed raw materials for later use;
(2) putting the uniformly mixed feed raw materials into a fermentation tank, adding beneficial microorganisms, and fermenting the feed raw materials by utilizing a beneficial microorganism fermentation enzymolysis technology to obtain fermented feed;
(3) adding at least three trace elements which are in the optimal feeding concentration range into the fermented feed, fully stirring and uniformly mixing, and adding water to ensure that the water content is kept to be 20-40% of the total weight of the feed to obtain a prefabricated feed;
(4) and (3) placing the prefabricated material in an extrusion granulator for extrusion forming to obtain the standard stichopus japonicus feed.
4. The method for tracing and constructing a apostichopus japonicus origin place according to claim 3, wherein the at least three trace elements are added in different combinations or in different proportions within the optimal feeding concentration range; the addition mode of the trace elements is direct addition or microorganism enrichment.
5. The method as claimed in claim 3, wherein the beneficial microorganism is one or a combination of two or more of yeast, probiotic bacillus, clostridium butyricum, lactobacillus, bifidobacterium or actinomycetes, and the amount of the beneficial microorganism is 0.2-1% of the total weight of the feed.
6. The method for constructing the stichopus japonicus origin tracing method according to claim 3, wherein the fermentation condition is fermentation at room temperature for 20-28 h.
7. Use of the method according to any one of claims 1 to 6 for tracing the origin of stichopus japonicus, for identifying the origin, quality and authenticity of products of stichopus japonicus.
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