Determination method of goat milk oligosaccharides
A determination method and technique of milk oligosaccharide, which are applied in the field of separation and determination of goat milk oligosaccharide, can solve the problems of increased operation difficulty, difficulty in separating low-content components, difficulty in qualitative determination, etc., and achieve the effects of stable results and simple and feasible determination method.
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Embodiment 1
[0058] Step 1, defatting and deproteinizing the goat milk. Centrifuge goat milk at 4°C and 10,000 rpm for 30 minutes to remove the fat in the upper layer and a small amount of protein at the bottom, take out the middle layer and add absolute ethanol at a ratio of 1:2 by volume, react at -20°C for 4 hours, and the reaction is complete Afterwards, centrifuge at 4000×g for 30 min at 4° C., take the supernatant, concentrate and dry in a vacuum concentrator, and obtain the crude goat milk oligosaccharide.
[0059] Step 2, purifying the crude goat milk oligosaccharide by using a Sephadex-G10 gel column to remove lactose. First, place Sephadex-G10 gel in a beaker and soak it in ultrapure water for 12 hours, remove air bubbles for 2 hours, and then add it to the chromatography column. Prepress at mL / min for 12h. The goat milk oligosaccharide crude product was loaded onto a Sephadex-G10 gel column, and ultrapure water was used as the eluent at a flow rate of 2.5mL / min. Sulfuric acid...
Embodiment 2
[0065] Step 1, defatting and deproteinizing the goat milk. Centrifuge goat milk at 4°C and 10,000 rpm for 30 minutes to remove the fat in the upper layer and a small amount of protein at the bottom, take out the middle layer and add absolute ethanol at a ratio of 1:2 by volume, react at -20°C for 4 hours, and the reaction is complete Afterwards, centrifuge at 4000×g for 30 min at 4° C., take the supernatant, concentrate and dry in a vacuum concentrator, and obtain the crude goat milk oligosaccharide.
[0066] Step 2, purifying the crude goat milk oligosaccharide by using a Sephadex-G10 gel column to remove lactose. First, place Sephadex-G10 gel in a beaker and soak it in ultrapure water for 12 hours, remove air bubbles for 2 hours, and then add it to the chromatography column. Prepress at mL / min for 12h. The goat milk oligosaccharide crude product was loaded onto a Sephadex-G10 gel column, and ultrapure water was used as the eluent at a flow rate of 2.5mL / min. Sulfuric acid...
Embodiment 3
[0072] Step 1, defatting and deproteinizing the goat milk. Centrifuge goat milk at 4°C and 10,000 rpm for 30 minutes to remove the fat in the upper layer and a small amount of protein at the bottom, take out the middle layer and add absolute ethanol at a ratio of 1:2 by volume, react at -20°C for 4 hours, and the reaction is complete Afterwards, centrifuge at 4000×g for 30 min at 4° C., take the supernatant, concentrate and dry in a vacuum concentrator, and obtain the crude goat milk oligosaccharide.
[0073] Step 2, purifying the crude goat milk oligosaccharide by using a Sephadex-G10 gel column to remove lactose. First, place Sephadex-G10 gel in a beaker and soak it in ultrapure water for 12 hours, remove air bubbles for 2 hours, and then add it to the chromatography column. Prepress at mL / min for 12h. The goat milk oligosaccharide crude product was loaded onto a Sephadex-G10 gel column, and ultrapure water was used as the eluent at a flow rate of 2.5mL / min. Sulfuric acid...
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