Method for separating 5-methyltetrahydrofolate and improving stability, and used wall material

A methyltetrahydrofolate and stability technology, which is applied to medical preparations with non-active ingredients, medical preparations containing active ingredients, and pharmaceutical formulas. It can solve the problems of slow separation speed and light influence, and achieve good results. Excellent effects of protection, bioavailability improvement, and embedding effect

Active Publication Date: 2020-04-24
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, silica gel column is mostly used for the separation of folic acid, but because the silica gel column needs to be activated by silica gel, and then perform operations such as packing, based on the instability of folic acid, separation using silica gel is inevitably affected by light, and the separation speed is slow.

Method used

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  • Method for separating 5-methyltetrahydrofolate and improving stability, and used wall material
  • Method for separating 5-methyltetrahydrofolate and improving stability, and used wall material
  • Method for separating 5-methyltetrahydrofolate and improving stability, and used wall material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1, a kind of method for isolating 5-methyltetrahydrofolate from Lactobacillus sake fermented liquid, carries out following steps successively:

[0053] 1), the preparation of bacteria slime:

[0054] Get a ring of activated Lactobacillus sake (L-sakei) in MRS liquid medium, 37 ℃ culture 24h; Get the culture solution obtained and inoculate in MRS liquid medium according to the inoculum size of 3% (volume %) at 37 ℃ Cultivate for 24 hours; then centrifuge (rotate at 10,000 rpm for 10 minutes), remove the supernatant, and obtain the sludge;

[0055] MRS liquid medium: peptone 10g, yeast powder 5g, glucose 20g, beef extract 10g, sodium acetate 5g, dipotassium hydrogen phosphate 2g, diamine hydrogen citrate 2g, Tween-80 1mL, magnesium sulfate 0.58g, manganese sulfate 0.25g , pH7.0, dilute to 1L with deionized water. Autoclave, 121°C, 15min.

[0056] Preparation of MRS solid medium: add 2% agar on the basis of MRS liquid medium; namely:

[0057] Peptone 10g, ye...

Embodiment 2

[0075] Embodiment 2, the preparation of glycosylation reaction product (wall material), carries out following steps successively:

[0076] 1), casein and chitosan oligosaccharide (1:1w / w), dissolved in phosphate buffer solution (0.2mol / L, pH8.0), so that the concentration of casein is 2mg / mL;

[0077] About 50ml of the mixed solution was placed on a magnetic stirrer and stirred evenly, and synthesized by a dual-frequency ultrasonic microwave ultraviolet combined catalytic synthesizer; ultrasonic power 300w, microwave power 450w, time 20min, temperature 100°C.

[0078] The degree of grafting of casein is 26.21%. The detection method of the degree of grafting is in accordance with the conventional OPA detection method.

[0079] 2), the obtained product is put into a dialysis bag (Mr=8,000~14,000D), and dialyzed in deionized water for (24±2)h to remove unbound product, and the dialysate (deionized water) is changed every 12h, and the Freeze-dry the retentate in the dialysis bag...

Embodiment 3

[0110] Embodiment 3, the method for improving the stability of 5-methyltetrahydrofolate, comprises the following steps:

[0111] The glycosylation reaction product (obtained in Example 2) was dissolved in 0.2 mol / L phosphate buffer solution (pH=7), the concentration of the glycosylation product was 1 mg / mL, and it was placed on a magnetic stirrer for stirring Evenly, then filter, take 4mL from the filtrate and place it in a 5mL sample bottle, then add 100μg of 5-methyltetrahydrofolate, 2 Under the protection of 25°C, seal and avoid light and mix well; get 5-methyltetrahydrofolate microcapsules.

[0112] The embedding rate of casein glycosylation complex on 5-methyltetrahydrofolate reached 75.29%.

[0113]

[0114] Comparative Example 3-1. The amount of 5-methyltetrahydrofolate added in Example 3 was changed from 100 μg to 50 μg, 150 μg, 200 μg, and 250 μg respectively, and the rest were the same as in Example 3.

[0115] Carry out the following experiments respectively: ...

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Abstract

The invention discloses a preparation method of a wall material for improving the stability of 5-methyltetrahydrofolate. The preparation method comprises the following steps: dissolving casein and chitosan oligosaccharide in a phosphate buffer solution of which the pH value is 7.0-10.0 for synthesis; dialyzing an obtained product in deionized water, drying, and carrying out glycosylation reaction,so that an obtained glycosylation reaction product can be used as a wall material for improving the stability of 5-methyltetrahydrofolate. The invention further provides a method for improving the stability of 5-methyltetrahydrofolate. 5-methyltetrahydrofolate is embedded by utilizing the wall material. The invention further provides a method for separating 5-methyltetrahydrofolate from lactobacillus sake fermentation liquor. The method comprises the following steps: enabling the fermentation liquor to pass through a C18 solid-phase extraction column, and eluting with ultrapure water; therefore, separation of 5-methyltetrahydrofolate from the fermentation liquid is realized.

Description

technical field [0001] The invention belongs to the field of food engineering, and in particular relates to a method for improving the stability of 5-methyltetrahydrofolate and wall materials used, and also relates to a method for separating 5-methyltetrahydrofolate from fermentation liquid. Background technique [0002] Folic acid is a water-soluble vitamin, mainly 5-methyltetrahydrofolate in the human body; its property is unstable, easy to decompose when exposed to light, and storage at room temperature will greatly destroy folic acid, and folic acid plays an important role in biological life activities, especially pregnant women . So folic acid needs to be microencapsulated. [0003] At present, silica gel column is mostly used for the separation of folic acid, but because the silica gel column needs to be activated by silica gel, and then perform operations such as packing, based on the instability of folic acid, separation using silica gel is inevitably affected by li...

Claims

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Application Information

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IPC IPC(8): A61K9/50A61K31/519A61K47/36A61K47/42C07D475/04A61P3/02
CPCA61K9/5052A61K9/5036A61K31/519C07D475/04A61P3/02
Inventor 熊春华齐健沈忱翁凡舒王小青钟晶莹陆海霞修丽丽顾青
Owner ZHEJIANG GONGSHANG UNIVERSITY
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