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Method for extracting biotin with molasses as raw material and detection method

A technology of biotin and molasses, which is applied in the preparation of test samples and the measurement of color/spectral characteristics, etc. It can solve the problems of time-consuming operation, large amount of solvent, and difficulty in biotin extraction and purification, and achieve the effect of simple operation

Inactive Publication Date: 2014-07-16
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, to the detection of biotin in molasses, microbial method and high-performance liquid chromatography are the most commonly used, but there are following defects: microbial method will bring larger detection error because most strains are not specific, and The whole operation process is time-consuming and laborious
High-performance liquid chromatography requires a large investment, a large amount of solvent, and high cost. Because molasses is thick, dark in color, and contains many impurities, sample processing, that is, the extraction and purification of biotin, is difficult, and high-performance liquid chromatography is difficult for samples. The purity requirements are relatively high, and it is not suitable to use this method for detection of molasses samples with complex components, high pigment content, and large interference from impurities.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 takes molasses as raw material to extract biotin

[0021] Take 10g of molasses, add 100ml of 80% ethanol (V / V) solution, stir evenly, put it in 40KHz, power 300W, temperature 60 ℃ under the condition of ultrasonic for 30 minutes. The molasses gradually changes from liquid to block, and the molasses block is taken out, and the ethanol solution is quickly cooled to below 5°C, and then the ethanol solution is poured out to obtain the remaining residue.

[0022] Combine the molasses block and the residue, rinse with the prepared ethanol solution of the same concentration at 60°C, and pour off the ethanol solution after cooling to below 5°C to obtain the molasses block.

[0023] Add 100ml of 95% ethanol to the molasses slag, at 20KHz, 300W, and a temperature of 55°C, the ultrasonic probe penetrates 3cm below the liquid surface. The interval is 5 seconds, and the ultrasonic extraction is continued for 30 minutes under the setting of ultrasonic 3 minutes. The mo...

Embodiment 2

[0028] Embodiment 2 takes molasses as raw material to extract biotin

[0029] Take 15g of molasses, add 150ml of 95% ethanol solution (V / V), stir evenly, and ultrasonicate for 20 minutes at 40KHz, power 500W, and temperature 70°C. The molasses gradually changes from liquid to block, take out the molasses block, and quickly cool the remaining ethanol solution to above 5°C, then pour out the ethanol solution to obtain a small amount of remaining residue.

[0030] Combine the molasses block and the residue, rinse with the prepared ethanol solution of the same concentration at 70°C, pour off the ethanol solution when it is cooled to below 5°C, and obtain the molasses residue block.

[0031] Add 150ml of absolute ethanol to the molasses residue, at 20KHz, 500W, and a temperature of 65°C, the ultrasonic probe penetrates 3cm below the liquid surface. Continuous ultrasonic extraction was performed for 20 minutes at intervals of 5 seconds and 3 minutes. The molasses block is powdered...

Embodiment 3

[0035] Embodiment 3 takes molasses as raw material to extract biotin

[0036] Take 12g of molasses, add 120ml of 85% ethanol solution (V / V), stir evenly, place in 40KHz, power 400W, temperature 65 ℃ and sonicate for 25 minutes. During the ultrasonic process, the molasses gradually changed from liquid to block, and the molasses block was taken out, and the remaining ethanol solution was quickly cooled to above 5°C, and then the ethanol solution was poured out to obtain a small amount of remaining residue. Combine the molasses block and residue, and rinse with the prepared ethanol solution of the same concentration at 65°C. Pour off the ethanol solution when cooled to below 5°C to obtain molasses residue.

[0037] Add 120ml of 95% ethanol to the molasses slag, at 20KHz, 400W, and a temperature of 60°C, the ultrasonic probe penetrates 3cm below the liquid surface. The interval is 5 seconds, and the ultrasonic extraction is continued for 25 minutes under the setting of ultrasoni...

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Abstract

The invention discloses a method for extracting biotin with molasses as a raw material. The method is characterized by using molasses as a raw material, adding ethanol, carrying out 40KHz ultrasonic treatment to obtain molasses clinkers, adding ethanol, carrying out 20KHz ultrasonic treatment and carrying out extraction with N,N-dimethylformamide in a cooperative manner, thus finally obtaining a sample solution containing biotin. The invention also provides a thin layer chromatography (TLC) scanning detection method of biotin. The detection method comprises the following steps: (1) dropping the sample solution to be detected on a silica gel plate, adding a developer, developing the sample solution to be detected in a developing cylinder, and then taking out the thin layer plate and putting the thin layer plate in a fume hood to be aired; (2) spray-dyeing the developed and aired plate with a dyeing solution to obtain spots, in obvious contrast to the background, of standard substances and samples; (3) carrying out TLC scanning by utilizing a thin layer scanner in the absorption wavelength of 530nm, and computing the mass concentration of biotin in the sample solution. The methods have the advantages of simplicity, quickness, accuracy, reliability, stability and the like.

Description

technical field [0001] The invention relates to a method for extracting biotin from molasses as a raw material and a detection method for biotin. Background technique [0002] Biotin is one of the essential vitamins in life activities. It is a cofactor of many enzymes in the body, participates in the carboxylation, decarboxylation and dehydrogenation reactions of the body, and participates in the metabolism of sugar, fat and protein. Biotin has been widely used in food, chemical industry, medicine, animal husbandry, biological fermentation and other fields. In the production of glutamic acid fermentation, the control of the amount of biotin directly affects the growth, proliferation, metabolism, cell wall, cell membrane permeability and the rate of glutamic acid production of the producing bacteria, because most glutamic acid bacteria are Biotin-deficient, so the content of biotin plays a vital role in the production, so it is of great significance to carry out research on ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N21/25
Inventor 刘代成
Owner SHANDONG NORMAL UNIV
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