A method for the determination of selenium speciation in selenium-enriched plant dry powder
A technology for selenium-enriched plants and dry powder, which is applied in the field of analysis and testing, can solve the problems of high cost of protease, selenium form conversion, long reaction time, etc., and achieves the effects of high extraction efficiency, short time and simple operation.
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Embodiment 1
[0035] (1) Dissolve 0.1 g of dry powder of rice chestnuts in 5 mL of deionized water, control the temperature at 40°C, and ultrasonicate at 40KHz for 15 minutes. After the ultrasonication is completed, centrifuge the obtained mixture at 4000 rpm for 15 minutes and take the supernatant to obtain supernatant 1;
[0036] (2) Dissolve 1 mg of sodium dodecyl sulfate (SDS) in 7 mL of Tris-HCL with a pH of 7.5, dissolve the precipitate after centrifugation in all steps (1), and add 10 mg of flavor protease (Flavourzyme500MG, Novozymes Biotechnology Co., Ltd.) was stirred for 4 hours, and then 10 mg of protease E (Pronase E, Type XIV, ≥3.5 units / mg, Sigma-aldrich Company) was stirred for 4 hours, and the temperature of the enzyme was 40 °C. After solution, centrifuge at 4000rpm for 15min to take the supernatant to obtain supernatant 2;
[0037] (3) Take the centrifuged precipitate in step (2) and repeat the operation in step (2) for enzymolysis. After enzymolysis, centrifuge at 4000rp...
Embodiment 2
[0048] (1) Dissolve 0.1g of dry cabbage powder in 5mL of deionized water, control the temperature at 40°C, and ultrasonicate at 40KHz for 25min. After the ultrasonication is completed, centrifuge the obtained mixture at 4000rpm for 15min and take the supernatant to obtain supernatant 1;
[0049] (2) Dissolve 1 mg of SDS in 7 mL of Tris-HCL with pH 7.5, dissolve the precipitate after centrifugation in step (1), first add 10 mg of flavor protease and stir for 4 hours, then add 10 mg of protease E and stir Enzyme hydrolysis for 4 hours, the temperature of enzymolysis is 40°C, after enzymolysis, centrifuge at 4000 rpm for 15 minutes to take the supernatant, and obtain supernatant 2;
[0050] (3) Take the centrifuged precipitate in step (2) and repeat the operation in step (2) for enzymolysis. After enzymolysis, centrifuge at 4000rpm for 15min to take the supernatant to obtain supernatant 3;
[0051] (4) Collect the supernatant obtained in each step and filter it through a 0.45 μm ...
Embodiment 3
[0053] (1) Dissolve 0.1g of broccoli dry powder in 7mL of deionized water, control the temperature at 40°C, and ultrasonicate at 40KHz for 15min. After the ultrasonication is completed, centrifuge the obtained mixture at 4000rpm for 15min and take the supernatant to obtain supernatant 1;
[0054] (2) Dissolve 1 mg of SDS in 7 mL of Tris-HCL with pH 7.5, dissolve the precipitate after centrifugation in step (1), first add 10 mg of flavor protease to it and stir for 5 hours, then add 10 mg of protease E and stir Enzymolysis was performed for 5 hours at a temperature of 40°C. After enzymolysis, centrifuge at 4000 rpm for 15 minutes to obtain supernatant 2;
[0055] (3) Take the centrifuged precipitate in step (2) and repeat the operation in step (2) for enzymolysis. After enzymolysis, centrifuge at 4000rpm for 15min to take the supernatant to obtain supernatant 3;
[0056] (4) Collect the supernatant obtained in each step and filter it through a 0.45 μm nylon filter head. The con...
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