Method for determination of form of selenium in selenium-rich plant material

Abstract

The invention discloses a method for determination of the form of selenium in a selenium-rich plant material. The method provided by the invention is characterized by comprising the following steps of 1, a selenium-rich plant material pretreatment process comprising carrying out extraction of a crushed selenium-rich plant material sample by an extraction buffer Tris-HCl, after the extraction, adding cellulase into the extract, carrying out wall breaking of cells of the selenium-rich plant material sample, orderly adding effective amounts of protease K and protease XIV into the extract, and carrying out hydrolysis extraction to obtain a solution of a sample needing to be detected, and 2, carrying out determination processes respectively on a solution of selenium having a standard form and the solution of the sample needing to be detected by the combination of the hydride generation-atomic fluorescence spectroscopy technology and the high performance liquid chromatography. The method provided by the invention is simple, can be realized easily, adopts nontoxic reagents and does not produce pollution.

Description

technical field

[0001] The invention relates to a method for determining the form of selenium in a selenium-rich plant material, in particular to a method for measuring the form of selenium in a selenium-rich plant material after pretreatment. Background technique

[0002] Selenium is one of the essential trace elements for the human body and a component of glutathione peroxidase (GSH-Px), which has the functions of anti-oxidation, anti-cancer, anti-aging, improving human immunity, and protecting cardiovascular and cerebrovascular ( Rotruck J T, Pope A L, Ganther H E, et al Selenium: biochemical role as a component of glutathione peroxidase. Science, 1973, 179, 588-590.). There are two-thirds of selenium-deficient areas in my country, and one-third of them are severely selenium-deficient areas. Selenium deficiency in humans and animals can cause Keshan disease, Kaschin-Beck disease, white muscle disease of animals, etc. In addition, some cancers, endemic thyroid disorders a...

Examples

Embodiment 1

[0057] Example 1 Determination of Selenium Species in Selenium-enriched Corn Stalks

[0058] The steps of the pretreatment method for the determination of selenium speciation in selenium-enriched corn stalks were optimized. The optimization process and results are as follows:

[0059] Weigh 0.2g selenium-enriched corn stalk sample into a 15mL plastic centrifuge tube, add 5mL extraction buffer Tris-HCl, ultrasonically extract for 20min, first add 40mg of cellulase, 50℃, shake at constant temperature for 1h, then add 0.5mg of proteinase K , 50°C, constant temperature shaking culture for 8 hours, then add 0.5 mg protease XIV, 37°C, constant temperature shaking culture for 8 hours, then at 4°C, rotate at 10000 rpm, centrifuge for 30 minutes, and pass the supernatant through a 0.22 μm filter membrane to prepare Test solution on the machine. Set the liquid chromatography conditions as mobile phase (40mmol / L(NH4)2HPO4, pH 6.0); flow rate 1mL / min; injection volume 100μL; peristaltic ...

Embodiment 2

[0078] Selenium speciation determination in embodiment 2 selenium-enriched wheat seedlings

[0079] Weigh 0.2g sample into a 15mL plastic centrifuge tube, add 5mL extraction buffer Tris-HCl, ultrasonically extract for 30min, add 40mg cellulase, 50°C, shake at constant temperature for 1h, then add 1mg proteinase K, shake at 50°C at constant temperature Cultivate for 12 hours, then add 1 mg of protease XIV, 37°C, shake at constant temperature for 12 hours, then centrifuge at 10,000 rpm for 30 minutes at 4°C, pass the supernatant through a 0.22 μm filter membrane, and measure the content of organic selenium on a machine. Other conditions are the same as in Example 1, and the results of the determination of the sample and the recovery of standard addition are shown in the following table.

[0080] Table 5 Optimization condition screening and effect

[0081]

[0082]It can be seen from Table 5 that in the pretreatment method of selenium-enriched wheat seedling selenium speciati...

technical field

[0001] The invention relates to a method for determining the form of selenium in a selenium-rich plant material, in particular to a method for measuring the form of selenium in a selenium-rich plant material after pretreatment. Background technique

[0002] Selenium is one of the essential trace elements for the human body and a component of glutathione peroxidase (GSH-Px), which has the functions of anti-oxidation, anti-cancer, anti-aging, improving human immunity, and protecting cardiovascular and cerebrovascular ( Rotruck J T, Pope A L, Ganther H E, et al Selenium: biochemical role as a component of glutathione peroxidase. Science, 1973, 179, 588-590.). There are two-thirds of selenium-deficient areas in my country, and one-third of them are severely selenium-deficient areas. Selenium deficiency in humans and animals can cause Keshan disease, Kaschin-Beck disease, white muscle disease of animals, etc. In addition, some cancers, endemic thyroid disorders a...

Examples

Embodiment 1

[0057] Example 1 Determination of Selenium Species in Selenium-enriched Corn Stalks

[0058] The steps of the pretreatment method for the determination of selenium speciation in selenium-enriched corn stalks were optimized. The optimization process and results are as follows:

[0059] Weigh 0.2g selenium-enriched corn stalk sample into a 15mL plastic centrifuge tube, add 5mL extraction buffer Tris-HCl, ultrasonically extract for 20min, first add 40mg of cellulase, 50℃, shake at constant temperature for 1h, then add 0.5mg of proteinase K , 50°C, constant temperature shaking culture for 8 hours, then add 0.5 mg protease XIV, 37°C, constant temperature shaking culture for 8 hours, then at 4°C, rotate at 10000 rpm, centrifuge for 30 minutes, and pass the supernatant through a 0.22 μm filter membrane to prepare Test solution on the machine. Set the liquid chromatography conditions as mobile phase (40mmol / L(NH4)2HPO4, pH 6.0); flow rate 1mL / min; injection volume 100μL; peristaltic ...

Embodiment 2

[0078] Selenium speciation determination in embodiment 2 selenium-enriched wheat seedlings

[0079] Weigh 0.2g sample into a 15mL plastic centrifuge tube, add 5mL extraction buffer Tris-HCl, ultrasonically extract for 30min, add 40mg cellulase, 50°C, shake at constant temperature for 1h, then add 1mg proteinase K, shake at 50°C at constant temperature Cultivate for 12 hours, then add 1 mg of protease XIV, 37°C, shake at constant temperature for 12 hours, then centrifuge at 10,000 rpm for 30 minutes at 4°C, pass the supernatant through a 0.22 μm filter membrane, and measure the content of organic selenium on a machine. Other conditions are the same as in Example 1, and the results of the determination of the sample and the recovery of standard addition are shown in the following table.

[0080] Table 5 Optimization condition screening and effect

[0081]

[0082]It can be seen from Table 5 that in the pretreatment method of selenium-enriched wheat seedling selenium speciati...

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