CN108619166A - A kind of nanosizing selenium-rich asparagus juice and preparation method thereof - Google Patents

Abstract

A nanometerized selenium-enriched asparagus juice, which is a kind of asparagus juice loaded with selenized nanoparticles with a particle size of 70-100nm on asparagus saponin, and its components are: 3-5 parts of saponin in asparagus juice, 4-6 parts of sodium selenite; the preparation method of the above-mentioned nano-selenium-enriched asparagus juice is mainly to react the concentrated asparagus juice, Na ₂ SeO ₃ and dilute nitric acid solution at 50-90°C for 6-10 hours in proportion, and pass the asparagus The asparagus saponin in the asparagus juice undergoes an esterification reaction with Na ₂ SeO ₃ , Se=O replaces the ‑OH in the saponin to generate selenite, and selenium is introduced into the asparagus juice, and then the pH is adjusted with anhydrous sodium carbonate to 4-7, centrifuge, and dialyze the residual Na ₂ SeO ₃ to obtain nanometerized selenium-enriched asparagus juice. The invention introduces the trace element selenium into the asparagus juice to make a nano selenium structure, greatly improves the bioavailability of the selenium, and can reduce its side effects.

Description

A kind of nano selenium-enriched asparagus juice and preparation method thereof

Technical field The present invention relates to an asparagus juice and a preparation method thereof.

Background Art Selenium is an essential trace element for the human body. It has the functions of anti-cancer, anti-oxidation, enhancing human immunity, antagonizing harmful heavy metals, and regulating protein synthesis. Since selenium cannot be synthesized in the human body, and there is no organ to store selenium, the selenium needed by the human body must be obtained from food. Selenium deficiency in the human body can cause a variety of diseases, such as cancer, cardiovascular disease, liver disease, cataract, pancreatic disease, diabetes, reproductive system diseases and other symptoms. China is a big selenium-deficient country, and the per capita daily selenium intake in most parts of China is only between 50-250 micrograms. Two-thirds of China's population is seriously deficient in selenium, which is manifested in a serious shortage of selenium intake. Therefore, it is necessary to develop a preparation for supplementing the trace element selenium needed by the human body.

my country is a big producer of asparagus. Asparagus is rich in vitamin B, vitamin A, folic acid, selenium, iron, manganese, zinc and other trace elements and various amino acids necessary for the human body. It has the effect of regulating the body's metabolism and improving the body's immunity. In addition, asparagus is rich in active substances, which can stimulate the body's immune-suppressing carcinogens and accelerate detoxification. It can also prevent the division and growth of cancer cells, even reverse the cancer cells, promote the formation of antibodies, and improve the resistance to cancer. ; In addition, the folic acid and nucleic acid contained in it can effectively regulate the balance of the body and improve immunity.

In traditional production, selenium-containing fertilizers are applied during the growth of asparagus. Although asparagus can contain selenium, the bioavailability of selenium is not high.

SUMMARY OF THE INVENTION The object of the present invention is to provide a nano-selenized asparagus juice that greatly improves the bioavailability of selenium without increasing the amount of selenium and its preparation method. The present invention mainly uses asparagus juice as a raw material, through the esterification reaction between asparagus saponin in asparagus juice and Na ₂ SeO ₃ , Se=O replaces -OH in saponin to generate selenite, and then removes residual Na ₂ SeO ₃ dialyzed to obtain asparagus juice rich in nano-selenium.

The nano-selenium-enriched asparagus juice of the present invention is a kind of asparagus juice in which selenized nanoparticles with a particle size of 70-100 nm are loaded on asparagus saponin, and its components are asparagus juice in terms of mass fraction: 3-5 parts of saponin in asparagus juice, sub 4-6 parts of sodium selenate.

The preparation method of above-mentioned nanometerized selenium-enriched asparagus juice is as follows:

(1) Add sodium selenite to asparagus juice, and ultrasonically oscillate for 2 to 10 minutes to promote the dissolution of sodium selenite. Adding a dilute nitric acid solution with a concentration of 0.2% to 0.7% by volume to the asparagus juice solution under the condition of magnetic stirring at ~60rpm, and the dropping speed is 1 to 5 seconds/drop;

(2) reacting the mixed solution of step (1) in a water bath for 6 to 10 hours under the condition of 30 to 70 r/min magnetic stirring, and the temperature of the water bath is 50 to 90° C.;

(3) Cool the solution in step (2) to room temperature, adjust the pH to 4-7 with anhydrous sodium carbonate, centrifuge the solution at 6000-10000 r/min, dialyze the supernatant in deionized water, and dialyze the solution every 1 Change the water every 3 hours, take out 1-5mL dialysate and use ascorbic acid to detect the residual amount of free Na ₂ SeO ₃ , stop the dialysis when no red color appears, and pass the obtained solution through a 0.2 μm sterile microporous membrane to sterilize , to prepare nano-selenium-enriched asparagus juice.

Compared with the prior art, the present invention has the following advantages

1. Compared with traditional production, by applying selenium-containing fertilizers during the growth of asparagus, the bioavailability of selenium is greatly improved without increasing the amount of selenium.

2. The selenium in the prepared asparagus juice is at the nanometer level and combined with asparagus saponin as a template, so it has good dispersion and is conducive to the absorption and utilization of the human body. The combination of natural biomolecule asparagus saponin with nano-selenium is also conducive to improving the biocompatibility of selenium sex.

3. The prepared nano-selenium-enriched asparagus juice not only has health effects such as improving immunity that other asparagus products have, but also has a killing effect on cancer cells.

4. The trace element selenium is introduced into the asparagus juice to form a nano-selenium structure, which can fully exert its biological efficacy and reduce its side effects.

Description of drawings

Fig. 1 is the transmission electron microscope picture of the nanometerization selenium-enriched asparagus juice that the embodiment of the present invention 1 makes

Fig. 2 is the infrared spectrogram of the nano-selenium-enriched asparagus juice prepared in Example 2 of the present invention.

Fig. 3 is a comparison chart of cytotoxicity analysis on Hela cells between nano-selenium-enriched asparagus juice and common asparagus juice prepared in Example 3 of the present invention.

Detailed ways

Example 1

Dissolve 0.200g Na ₂ SeO ₃ (purchased from Shandong West Asia Chemical Industry Co., Ltd.) into 6.820mL concentrated asparagus juice (purchased from Qinhuangdao Changsheng Nutrition and Health Technology Co., Ltd., the main component is asparagus saponin, content ≥ 2.2 g/100 ml) , ultrasonically oscillate for 2 minutes to dissolve Na ₂ SeO ₃ ; add 19.325 mL of dilute nitric acid solution with a concentration of 0.2% to the solution dropwise under the condition of 20 rpm magnetic stirring, and control the dropping rate to 1 second/drop; under the condition of 30 rpm magnetic stirring, The water bath was reacted for 6 hours, and the temperature of the water bath was 50°C; then the solution was cooled to room temperature, and the pH was adjusted to 4 with anhydrous sodium carbonate, and the solution was centrifuged at 6000 rpm, and the supernatant was dialyzed in deionized water, and changed every 1 hour. Take out 1 mL of the dialysate and use ascorbic acid to detect the residual amount of free Na ₂ SeO ₃ , and stop the dialysis when no red color appears; the obtained solution is sterilized by a 0.2 μm sterile microporous membrane to obtain nano-rich Selenium asparagus juice, stored in a refrigerator at 4°C.

As shown in Figure 1, it can be seen that selenium atoms are successfully combined into asparagus saponin, and its particle size is at the nanometer level, ranging from 70 to 100 nm.

Example 2

Dissolve 0.250g Na ₂ SeO ₃ (purchased from Shandong West Asia Chemical Industry Co., Ltd.) into 9.090mL concentrated asparagus juice (purchased from Qinhuangdao Changsheng Nutrition and Health Technology Co., Ltd., the main component is asparagus saponin, content ≥ 2.2 g/100 ml) , ultrasonically vibrate for 6 minutes to dissolve Na ₂ SeO ₃ ; add 20.20 mL of dilute nitric acid solution with a concentration of 0.5% to the solution dropwise under the condition of 40 rpm magnetic stirring, and control the dropping speed to 3 seconds/drop; Under the condition of water bath reaction for 8 hours, the water bath temperature is 70 ℃; then the solution is cooled to room temperature, the pH is adjusted to 6 with anhydrous sodium carbonate, the solution is centrifuged at 8000 rpm, and the supernatant is dialyzed in deionized water every 2 Change the water every hour, take out 3mL dialysate and use ascorbic acid to detect the residual amount of free Na ₂ SeO ₃ , and stop the dialysis when no red color appears; the obtained solution is sterilized by a 0.2 μm sterile microporous membrane to obtain nanometer Selenium-enriched asparagus juice.

The prepared nano-selenium-enriched asparagus juice and concentrated asparagus juice were put into a freeze-dryer to freeze-dry, and an infrared spectrometer was used to detect the asparagus juice rich in nano-selenium and concentrated asparagus juice, as shown in Figure 2. The waveform, absorption intensity and peak width of the asparagus juice rich in nano-selenium were basically the same as those of the unmodified asparagus juice, indicating that the structure of the modified product itself was not damaged and the original properties of the asparagus juice were maintained. The difference is that the infrared spectrum of the asparagus juice rich in nano-selenium shows an obvious characteristic absorption peak at 785.32cm ^-1 , which is attributed to the Se=O stretching vibration peak, which proves that sodium selenite is successfully combined into the asparagus juice .

Example 3

Dissolve 0.300g Na ₂ SeO ₃ (purchased from Shandong West Asia Chemical Industry Co., Ltd.) into 11.365mL concentrated asparagus juice (purchased from Qinhuangdao Changsheng Nutrition and Health Technology Co., Ltd., the main component is asparagus saponin, content ≥ 2.2 g/100 ml) In the solution, ultrasonically vibrate for 10 minutes to dissolve Na ₂ SeO ₃ ; add 21.78mL of 0.7% dilute nitric acid solution dropwise to the solution under the condition of 60rpm magnetic stirring, and control the dropping speed to 5 seconds/drop; React for 10 hours, the temperature of the water bath is 90 ° C; then cool the solution to room temperature, adjust the pH to 7 with anhydrous sodium carbonate, centrifuge the solution at 10,000 rpm, dialyze the supernatant in deionized water, and change it every 3 hours water, and take out 5mL of the dialysate to detect the residual amount of free Na ₂ SeO ₃ with ascorbic acid, and stop the dialysis when no red color appears; the resulting solution is sterilized by a 0.2 μm sterile microporous membrane to obtain nano-selenium-enriched Store the asparagus juice in a refrigerator at 4°C.

MTT experiment: Hela cells in the logarithmic growth phase were inoculated into a 96-well plate at a density of 5,000 cells per well, and the amount added to each well was 100 μL, and placed in a 37°C, 5% _CO2 constant temperature incubator for cultivation. And observe regularly, after 24 hours, give nano-selenium-enriched asparagus juice; each group of samples to be tested is filtered with a 0.2 μL filter membrane and diluted 10 times with PBS buffer, and each group is set with 3 duplicate holes, and nano-selenium-enriched asparagus juice is given 100 μL, and then continue to culture in a constant temperature incubator for 24 hours; bromide tetrazolium (MTT) solution, and continue to incubate for 4 hours; suck out the supernatant in each well, and add 150 μL of dimethyl sulfoxide (DMSO) respectively, and wait until the crystals are fully dissolved. The absorbance was measured, and the cell viability in the wells of each group of samples was calculated. As can be seen from Figure 3, compared with the concentrated asparagus juice without selenium, the asparagus juice rich in nano-selenium has obvious killing effect on Hela cells, and as the concentration of asparagus juice rich in nano-selenium increases, the The killing effect of Hela cells is also gradually enhanced.

Claims (8)

1. A nanoselenium-enriched asparagus juice, characterized in that: it is a kind of asparagus juice with a particle diameter of 70～100nm selenized nanoparticles loaded on the asparagus saponin, and its components are by mass fraction: in the asparagus juice 3-5 parts of saponin, 4-6 parts of sodium selenite. 2. the preparation method of the nanometerized selenium-enriched asparagus juice of claim 1, is characterized in that: (1) Add sodium selenite to the asparagus juice, ultrasonically vibrate, and drop it into the asparagus juice solution under the condition of magnetic stirring according to the volume ratio of asparagus juice and dilute nitric acid solution of 6-12:17-23 Add dilute nitric acid solution; (2) reacting the mixed solution of step (1) in a water bath under magnetic stirring conditions for 6 to 10 hours; (3) Cool the solution in step (2) to room temperature, adjust the pH to 4-7 with anhydrous sodium carbonate, centrifuge the solution, dialyze the supernatant in deionized water, and change the water every 1-3 hours , and take out 1 ~ 5mL dialysate and use ascorbic acid to detect the residual amount of free Na ₂ SeO ₃ , stop the dialysis when no red color appears, and pass the obtained solution through a 0.2 μm sterile microporous membrane to sterilize to obtain nano-rich Se asparagus juice. 3. The method for preparing nano-selenium-enriched asparagus juice according to claim 2, characterized in that: the ultrasonic vibration treatment in the step (1) uses ultrasonic waves for 2-10 minutes. 4. The method for preparing nano-selenium-enriched asparagus juice according to claim 2, characterized in that: the stirring speed in the step (1) is 20-60 rpm. 5. The preparation method of nano-selenium-enriched asparagus juice according to claim 2, characterized in that: the volume percent concentration of dilute nitric acid in the step (1) is 0.2% to 0.7%, and the dropping speed is 1 to 5 seconds /drop. 6. The method for preparing nano-selenium-enriched asparagus juice according to claim 2, characterized in that: the rotating speed of the medium magnetic stirring in the step (2) is 30-70 rpm. 7. The method for preparing nano-selenium-enriched asparagus juice according to claim 2, characterized in that: the temperature of the water bath in the step (2) is 50-90°C. 8. The method for preparing nano-selenium-enriched asparagus juice according to claim 2, characterized in that: the centrifugal speed in the step (3) is 6000-10000 rpm.