CN112649407A - Method for evaluating safety of organic selenium and inorganic selenium in agricultural products - Google Patents
Method for evaluating safety of organic selenium and inorganic selenium in agricultural products Download PDFInfo
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Abstract
The invention relates to a method for evaluating the safety of organic selenium and inorganic selenium in agricultural products. Adding an alkali solution into the selenium-rich agricultural product dry powder, oscillating in a constant temperature shaking table at 30-50 ℃ for 2-3h, centrifuging, filtering, collecting filtrate to obtain leaching liquor containing organic selenium, and measuring the concentration of the organic selenium in the obtained leaching liquor; preparing an aqueous solution containing inorganic selenium by taking sodium selenite as a source of the inorganic selenium, wherein the concentration of the inorganic selenium is similar to that of the organic selenium in the leaching liquor, respectively preparing an organic selenium culture solution and an inorganic selenium culture solution, and respectively culturing rooted wheat seedlings in glass beakers by taking a universal culture solution as a blank; and when the wheat seedlings grow to a two-leaf one-heart state, taking out the wheat seedlings, and measuring the content of malondialdehyde and proline in the wheat seedlings. The safety of organic selenium and inorganic selenium in the selenium-rich agricultural products is visually evaluated according to the contents of malondialdehyde and proline.
Description
Technical Field
The invention belongs to the technical field of agricultural production, and particularly relates to a method for contrastively evaluating the safety of organic selenium and inorganic selenium in agricultural products.
Background
Selenium is a trace element necessary for human bodies, but nearly one hundred thousand of people in the world are lack of selenium, and the lack of selenium can make people easily get cancers, keshan disease, Kaschin-Beck disease and the like, so the selenium supplement is particularly important. The international nutrition society recommends that adults take 50-200 mug of selenium element every day for the health of human bodies, in areas with severe selenium deficiency, people supplement selenium mainly eat inorganic selenium, namely selenium tablets (sodium selenite), however, the inorganic selenium has toxicity, and the direct large-amount supplement of the inorganic selenium can cause damage to human bodies. However, the selenium-rich product in the market is mixed, whether organic selenium is contained or inorganic selenium is simply combined with agricultural products, and whether a human body can safely supplement selenium cannot be known, so that the development of a novel evaluation method for detecting the safety difference between the organic selenium and the inorganic selenium in the agricultural products is very important.
Disclosure of Invention
The invention aims to provide a method which is simple and easy to operate and can intuitively evaluate the safety of organic selenium and inorganic selenium in selenium-rich agricultural products.
The technical scheme adopted by the invention is as follows: a method for evaluating the safety of organic selenium and inorganic selenium in agricultural products comprises the following steps:
1) cleaning selenium-rich agricultural products to be detected, drying and grinding to obtain selenium-rich agricultural product dry powder;
2) adding an alkali solution into the selenium-rich agricultural product dry powder, oscillating in a constant temperature shaking table at 30-50 ℃ for 2-3h, centrifuging, filtering the obtained supernatant, collecting filtrate to obtain leaching liquor containing organic selenium, and measuring the concentration of the organic selenium in the obtained leaching liquor;
3) using the leaching liquor with known organic selenium concentration obtained in the step 2) as a solventAdding K2SO4，Fe-EDTA，Ca(NO3)2，MgSO4·7H2O，KH2PO4，KCl，HBO3，ZnSO4，MnSO4·4H2O，CuSO4·5H2O and (NH)4)6Mo7O4Regulating pH to 6.5-7.5, and preparing organic selenium culture solution;
4) taking sodium selenite as a source of inorganic selenium, dissolving the sodium selenite in distilled water, and preparing an aqueous solution containing inorganic selenium, wherein the concentration of the inorganic selenium in the aqueous solution containing the inorganic selenium is equal to the concentration of organic selenium in a leaching solution +/-5 mu g/kg;
5) taking the aqueous solution with known inorganic selenium concentration prepared in the step 4) as a solvent, adding K2SO4，Fe-EDTA，Ca(NO3)2，MgSO4·7H2O，KH2PO4，KCl，HBO3，ZnSO4，MnSO4·4H2O，CuSO4·5H2O and (NH)4)6Mo7O4Adjusting pH to 6.5-7.5, and preparing inorganic selenium culture solution;
6) taking three 500ml glass beakers, respectively adding 500ml of organic selenium culture solution, 500ml of inorganic selenium culture solution and 500ml of general culture solution, respectively placing nylon nets above the glass beakers, respectively immersing rooted wheat seedling roots into the culture solution through nylon meshes, and supplementing the general culture solution according to the loss condition of the culture solution in the glass beakers every day;
7) and when the wheat seedlings grow to a two-leaf one-heart state, taking out the wheat seedlings, and measuring the content of malondialdehyde and proline in the wheat seedlings.
Preferably, in the method for evaluating the safety of organic selenium and inorganic selenium in agricultural products, in the step 2), the alkali solution is 0.1mol/L sodium hydroxide solution.
Preferably, in the method for evaluating the safety of organic selenium and inorganic selenium in agricultural products, in the step 2), 50mL of 0.1mol/L sodium hydroxide solution is added to every 1.0g of dry agricultural product powder.
Preferably, the agricultural product comprisesThe method for evaluating the safety of the organic selenium and the inorganic selenium comprises the following steps in step 3): the organic selenium culture solution per liter contains 0.75 × 10-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance is leaching liquor with known organic selenium concentration, and the pH value is adjusted to 6.5-7.5.
Preferably, in the method for evaluating the safety of organic selenium and inorganic selenium in agricultural products, in the step 5), the composition of the inorganic selenium culture solution is as follows: 0.75X 10 inorganic selenium culture solution/liter-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance is aqueous solution with known inorganic selenium concentration, and the pH is adjusted to 6.5-7.5.
Preferably, in the method for evaluating the safety of organic selenium and inorganic selenium in agricultural products, in step 6), the general culture solution comprises: 0.75X 10-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance of water, and the pH is adjusted to 6.5-7.5.
Preferably, in the method for evaluating the safety of organic selenium and inorganic selenium in agricultural products, in the step 6), the aperture of the nylon net is 2mm, and a root system of a wheat seedling is placed in each hole.
Preferably, the method for evaluating the safety of organic selenium and inorganic selenium in agricultural products comprises the following steps: selecting wheat seeds with full seeds, placing the wheat seeds in a culture dish which is fully soaked with ultrapure water gauze, and accelerating germination for 24 hours under the constant temperature and light-proof condition of 30 ℃.
The invention has the beneficial effects that:
1. according to the invention, the organic selenium in the agricultural products is leached by the alkali solution, so that the leaching rate is high.
2. According to the invention, the safety of the organic selenium and the inorganic selenium can be evaluated according to the influence result of the organic selenium culture solution and the inorganic selenium culture solution on the growth of wheat seedlings.
3. The invention can not only evaluate the difference of the safety of organic selenium and inorganic selenium, but also can be used as an index for whether selenium in selenium-rich agricultural products can harm human bodies.
Detailed Description
Preparation of general culture solution
The general culture solution comprises the following components: 0.75X 10-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10- 3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10- 7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance of water, and the pH is adjusted to 6.8. Used in the following examples.
Second, rooted wheat seedling
The cultivation method comprises the following steps: wheat seeds with full seeds are selected and placed in a culture dish which is soaked with ultrapure water gauze, and germination acceleration culture is carried out for 24 hours under the constant temperature and light-proof condition of 30 ℃, so as to obtain rooted wheat seedlings, which are used in the following examples.
Example 1
Mung bean sprouts are common agricultural products, are usually found on dining tables of every family, contain various plant proteins, amino acids and dietary fibers which are beneficial to human bodies, have low calorie, can prevent cancers in the aspects of a plurality of digestive tracts, effectively remove fat and cholesterol accumulated in blood vessel walls, and very accord with living habits of healthy diet of modern people. Selenium-rich mung bean sprouts also frequently appear in the market.
In the embodiment, selenium-rich mung bean sprouts are selected to evaluate the safety of organic selenium and inorganic selenium. Selecting 5 kinds of selenium-rich mung bean sprouts and 3 kinds of common mung bean sprouts for detection. The method comprises the following steps:
1. cleaning 5 kinds of selenium-rich mung bean sprouts and 3 kinds of common mung bean sprouts respectively, drying at 45 ℃, and grinding into powder to obtain mung bean sprout dry powder;
2. respectively adding 10g of mung bean sprout dry powder into 500mL of 0.1mol/L sodium hydroxide solution, oscillating for 2h in a constant temperature shaking table at 40 ℃, transferring to a centrifuge tube, centrifuging at the speed of 4000r/min for 20min at 4 ℃, filtering the obtained supernatant by using medium-speed filter paper, collecting filtrate, respectively obtaining organic selenium-containing selenium-rich mung bean sprout leaching liquor derived from 5 kinds of selenium-rich mung bean sprouts and common mung bean sprout leaching liquor derived from 3 kinds of common mung bean sprouts, and measuring the concentration of organic selenium in the obtained 5 kinds of organic selenium-containing selenium-rich mung bean sprout leaching liquor by using an atomic fluorescence photometer, wherein the results are shown in Table 1;
3. respectively using 5 known organic compounds obtained in step 2Extracting selenium-rich semen Phaseoli Radiati Germinatus extract with selenium concentration as solvent, and adding K2SO4，Fe-EDTA，Ca(NO3)2，MgSO4·7H2O，KH2PO4，KCl，HBO3，ZnSO4，MnSO4·4H2O，CuSO4·5H2O and (NH)4)6Mo7O4Adjusting pH to 6.8, and respectively preparing organic selenium culture solution. The organic selenium culture solution comprises the following components: the organic selenium culture solution per liter contains 0.75 × 10-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance is selenium-rich mung bean sprout leaching liquor with known organic selenium concentration, and the pH is adjusted to 6.8.
Respectively taking the 3 common mung bean sprout leaching liquor obtained in the step 2 as a solvent, and adding K2SO4，Fe-EDTA，Ca(NO3)2，MgSO4·7H2O，KH2PO4，KCl，HBO3，ZnSO4，MnSO4·4H2O，CuSO4·5H2O and (NH)4)6Mo7O4Adjusting the pH value to 6.8, and preparing common culture solutions respectively. The composition of the common culture solution is as follows: 0.75X 10 per liter of the common culture medium-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance of common mung bean sprout leaching liquor, and the pH is adjusted to 6.8.
4. Taking sodium selenite as a source of inorganic selenium, dissolving the sodium selenite in distilled water, preparing 5 kinds of aqueous solutions containing inorganic selenium, and enabling the 5 kinds of aqueous solutions containing the inorganic selenium to respectively correspond to the 5 kinds of organic selenium-rich mung bean sprout leaching liquor in the step 2 one by one, so that the concentration of the inorganic selenium in the aqueous solutions containing the inorganic selenium is equal to the concentration of organic selenium in the selenium-rich mung bean sprout leaching liquor +/-5 mug/kg;
5. taking the aqueous solution with known inorganic selenium concentration prepared in the step 4 as a solvent, and adding K2SO4，Fe-EDTA，Ca(NO3)2，MgSO4·7H2O，KH2PO4，KCl，HBO3，ZnSO4，MnSO4·4H2O，CuSO4·5H2O and (NH)4)6Mo7O4Adjusting pH to 6.8, and preparing inorganic selenium culture solution respectively. The inorganic selenium culture solution comprises the following components: 0.75X 10 inorganic selenium culture solution/liter-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10- 8mol/L(NH4)6Mo7O4The balance is aqueous solution with known inorganic selenium concentration, and the pH is adjusted to 6.5-7.5
6. Respectively adding 5 kinds of 500ml organic selenium culture solution, 5 kinds of 500ml inorganic selenium culture solution, 3 kinds of 500ml common culture solution and 500ml general culture solution into 14 glass beakers of 500ml, respectively placing nylon nets above the glass beakers, fixing the nylon nets by using rubber bands, wherein the aperture of each nylon net is 2mm, placing a wheat seedling root system into each hole, immersing the rooted wheat seedling root system into the culture solution through nylon meshes, and supplementing the general culture solution every day according to the loss condition of the culture solution in the glass beakers.
7. And when the wheat seedlings grow to a two-leaf one-heart state, taking out the wheat seedlings, and measuring the content of malondialdehyde and proline in the wheat seedlings. The results are shown in Table 1.
TABLE 1
| Selenium concentration (μ g/kg) | Malondialdehyde (mu g/g) | Proline (mug/g) | |
| Selenium-rich mung bean sprout sample 1 | 61.02 | 0.86 | 95 |
| Corresponding inorganic selenium water solution 1 | 60.00 | 0.92 | 100 |
| Selenium-rich mung bean sprout sample 2 | 40.17 | 0.90 | 95 |
| Corresponding inorganic selenium water solution 2 | 40.00 | 0.93 | 98 |
| Selenium-rich mung bean sprout sample 3 | 1304.46 | 1.13 | 133 |
| Corresponding inorganic selenium water solution 3 | 1300.00 | 1.28 | 146 |
| Selenium-rich mung bean sprout sample 4 | 152.91 | 0.99 | 104 |
| Corresponding inorganic selenium water solution 4 | 150.00 | 1.04 | 117 |
| Selenium-rich mung bean sprout sample 5 | 2959.37 | 1.31 | 162 |
| Corresponding inorganic selenium waterLiquid 5 | 2960.00 | 1.47 | 177 |
| Common mung Bean sprout sample 1 | 5.32 | 1.13 | 90 |
| Common mung Bean sprout sample 2 | 2.70 | 1.07 | 89 |
| Common mung Bean sprout sample 3 | 2.27 | 1.07 | 86 |
| General culture solution for control example | 0 | 1.08 | 86 |
When the content of malondialdehyde in wheat seedlings is more than 1.3 mu g/g and the content of proline is more than 140 mu g/g, selenium is considered to have harmful effect on the wheat seedlings and has no safety. As can be seen from Table 1, when the content of inorganic selenium is 1300 mug/kg, the content of malondialdehyde in wheat seedlings is close to 1.3 mug/g, but when the content of organic selenium is 2959.37 mug/kg, the content of malondialdehyde just exceeds 1.3 mug/g, and according to malondialdehyde index analysis, the safety range of organic selenium of the wheat seedlings is less than 2959.37 mug/kg, and the safety range of inorganic selenium is less than 1300.00 mug/kg; when the content of inorganic selenium is 1300 mu g/kg, the proline content in the wheat seedlings is higher than 140 mu g/g, but when the content of organic selenium is 1304.46 mu g/kg, the proline content of the wheat seedlings is lower than 140 mu g/g, and according to proline index analysis, the safety range of the organic selenium is larger than 1304.46 mu g/kg, and the safety range of the inorganic selenium is smaller than 1300 mu g/kg; by integrating the index analysis of malondialdehyde and proline, the upper limit of the safety range of organic selenium of wheat seedlings is 1304.46-2959.37 mug/kg, and the upper limit of the safety range of inorganic selenium is less than 1300 mug/kg. The conclusion that the organic selenium in the selenium-rich mung bean sprouts is safer than inorganic selenium can be obtained through the method, and the method is proved to be feasible.
Claims (8)
1. A safety evaluation method for organic selenium and inorganic selenium in agricultural products is characterized by comprising the following steps:
1) cleaning selenium-rich agricultural products to be detected, drying and grinding to obtain selenium-rich agricultural product dry powder;
2) adding an alkali solution into the selenium-rich agricultural product dry powder, oscillating in a constant temperature shaking table at 30-50 ℃ for 2-3h, centrifuging, filtering the obtained supernatant, collecting filtrate to obtain leaching liquor containing organic selenium, and measuring the concentration of the organic selenium in the obtained leaching liquor;
3) taking the leaching liquor with the known organic selenium concentration obtained in the step 2) as a solvent, and adding K2SO4，Fe-EDTA，Ca(NO3)2，MgSO4·7H2O，KH2PO4，KCl，HBO3，ZnSO4，MnSO4·4H2O，CuSO4·5H2O and (NH)4)6Mo7O4Regulating pH to 6.5-7.5, and preparing organic selenium culture solution;
4) taking sodium selenite as a source of inorganic selenium, dissolving the sodium selenite in distilled water, and preparing an aqueous solution containing inorganic selenium, wherein the concentration of the inorganic selenium in the aqueous solution containing the inorganic selenium is equal to the concentration of organic selenium in a leaching solution +/-5 mu g/kg;
5) taking the aqueous solution with known inorganic selenium concentration prepared in the step 4) as a solvent, adding K2SO4，Fe-EDTA，Ca(NO3)2，MgSO4·7H2O，KH2PO4，KCl，HBO3，ZnSO4，MnSO4·4H2O，CuSO4·5H2O and (NH)4)6Mo7O4Adjusting pH to 6.5-7.5, and preparing inorganic selenium culture solution;
6) taking three 500ml glass beakers, respectively adding 500ml of organic selenium culture solution, 500ml of inorganic selenium culture solution and 500ml of general culture solution, respectively placing nylon nets above the glass beakers, respectively immersing rooted wheat seedling roots into the culture solution through nylon meshes, and supplementing the general culture solution according to the loss condition of the culture solution in the glass beakers every day;
7) and when the wheat seedlings grow to a two-leaf one-heart state, taking out the wheat seedlings, and measuring the content of malondialdehyde and proline in the wheat seedlings.
2. The method for evaluating the safety of organic selenium and inorganic selenium in agricultural products of claim 1, wherein in the step 2), the alkali solution is 0.1mol/L sodium hydroxide solution.
3. The method for evaluating the safety of organic selenium and inorganic selenium in agricultural products as claimed in claim 2, wherein 50mL of 0.1mol/L sodium hydroxide solution is added to 1.0g of dry powder of agricultural products in step 2).
4. The method for evaluating the safety of organic selenium and inorganic selenium in agricultural products according to claim 1, wherein in the step 3), the composition of the organic selenium culture solution is as follows: the organic selenium culture solution per liter contains 0.75 × 10-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance is leaching liquor with known organic selenium concentration, and the pH value is adjusted to 6.5-7.5.
5. The method for evaluating the safety of organic selenium and inorganic selenium in agricultural products according to claim 1, wherein in the step 5), the composition of the inorganic selenium culture solution is as follows: 0.75X 10 inorganic selenium culture solution/liter-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10-3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance is aqueous solution with known inorganic selenium concentration, and the pH is adjusted to 6.5-7.5.
6. The method for evaluating the safety of organic selenium and inorganic selenium in agricultural products according to claim 1, wherein in the step 6), the composition of the universal culture solution is as follows: 0.75X 10-3mol/L K2SO4，1.1×10-4mol/L Fe-EDTA，2.0×10-3mol/L Ca(NO3)2，0.60×10-3mol/L MgSO4·7H2O，0.20×10-3mol/L KH2PO4，1.0×10- 3mol/L KCl，1.0×10-6mol/L HBO3，0.9×10-6mol/L ZnSO4，1.0×10-6mol/L MnSO4·4H2O，5.1×10-7mol/L CuSO4·5H2O，5.0×10-8mol/L(NH4)6Mo7O4And the balance of water, and the pH is adjusted to 6.5-7.5.
7. The method for evaluating the safety of organic selenium and inorganic selenium in agricultural products according to claim 1, wherein in the step 6), the aperture of the nylon net is 2mm, and a root of a wheat seedling is put into each hole.
8. The method for evaluating the safety of organic selenium and inorganic selenium in agricultural products according to claim 1, wherein the method for cultivating rooted wheat seedlings comprises the following steps: selecting wheat seeds with full seeds, placing the wheat seeds in a culture dish which is fully soaked with ultrapure water gauze, and accelerating germination for 24 hours under the constant temperature and light-proof condition of 30 ℃.
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