Method for separating unsaturated fatty acids from silkworm chrysalis oil based on composition complexation of reversible reaction

Abstract

The invention discloses a method for separating unsaturated fatty acids from silkworm chrysalis oil based on composition complexation of a reversible reaction. Combined separation is performed with a urea inclusion method and a beta-cyclodextrin inclusion method based on a reversible complex reaction, so that the total purity and total yield of unsaturated fatty acids such as alpha-linolenic acid, oleic acid, linoleic acid and the like in silkworm chrysalis oil can be increased to 86.5 percent and 91.1 percent, a separating process is easy and convenient to operate, the cost is low, and a theoretical basis and technical support are provided for future large-scale preparation of silkworm chrysalis oil unsaturated fatty acids and products thereof.

Description

Technical field

[0001] The invention belongs to the technical field of biochemical industry, and specifically relates to a reversible reaction-based combined complexation separation method for preparing high-purity unsaturated fatty acids (α-linolenic acid, linoleic acid and oleic acid) from silkworm pupa oil. Background technique

[0002] my country is a big country in the sericulture industry. As a by-product of silk reeling, silkworm pupa can produce nearly 200,000 tons of dried pupa each year, which is an important biomass oil resource. Taking silkworm pupa oil as an example, fat accounts for 9%-10% of fresh silkworm pupa, or 28%-30% of dry matter. Among them, saturated fatty acids account for about 20%, phospholipids and sterols account for about 4%, and unsaturated fatty acids account for about 75%. The unsaturated fatty acid of silkworm pupa oil is mainly composed of α-linolenic acid (about 30%), linoleic acid and oleic acid. It has many physiological functions such as lo...

Examples

Embodiment 1

[0025] The pupa mixed fatty acid and the saturated methanol solution of urea were stirred and mixed at a volume ratio of 1:1, clathrated at 0°C for 24h, the clathrate was cooled and crystallized, filtered, the filtrate obtained was extracted with petroleum ether, heated to reflux at 60°C, and the extract was washed with water , Vacuum degree of 0.001Mpa vacuum filtration, vacuum degree of 0.001Mpa, temperature of 90℃ to recover the solvent to obtain crude unsaturated fatty acid. The crude unsaturated fatty acid was dissolved in ethyl acetate to prepare a saturated solution, and then mixed with saturated β-cyclodextrin aqueous solution at a volume ratio of 1:5, heated in a water bath at 30°C for 0.1h, and shaken to obtain clathrates. Frozen at low temperature for 0.1h, filtered and dried to obtain β-cyclodextrin inclusion compound. The obtained clathrate is extracted with petroleum ether, heated to reflux at 60°C, and the extract is washed with water, and filtered under reduced ...

Embodiment 2

[0027] The mixed fatty acid of silkworm pupa and the saturated n-butanol solution of urea were stirred and mixed in a volume ratio of 1:10, and the inclusion compound was cooled and crystallized at 20°C for 1 h. The obtained filtrate was extracted with n-hexane, heated to reflux at 70°C, and washed with water. The extract is filtered under reduced pressure with a vacuum of 0.098Mpa, a vacuum of 0.098Mpa, and a temperature of 60°C to recover the solvent to obtain crude unsaturated fatty acids. The crude unsaturated fatty acid was dissolved in acetone to prepare a saturated solution, and then mixed with saturated β-cyclodextrin aqueous solution at a volume ratio of 1:15, heated in a water bath at 80°C for 24h, shaken to obtain clathrates, and frozen at 10°C for 24h. Filter and dry to obtain β-cyclodextrin inclusion compound. The obtained clathrate is extracted with petroleum ether, heated to reflux at 90°C, and the extract is washed with water, and the extract is filtered under r...

Embodiment 3

[0029] The pupa mixed fatty acid and the saturated ethanol solution of urea were stirred and mixed at a volume ratio of 1:2, clathrated for 2 hours at 4℃, the clathrate was cooled and crystallized, filtered, the filtrate obtained was extracted with petroleum ether, heated to reflux at 90℃, and the extract was washed with water , Vacuum degree of 0.090Mpa vacuum filtration, vacuum degree of 0.090Mpa, temperature 75℃ to recover the solvent to obtain crude unsaturated fatty acid. The crude unsaturated fatty acid was dissolved in ethanol to prepare a saturated solution, and then mixed with saturated β-cyclodextrin aqueous solution at a volume ratio of 1:9, heated in a water bath at 60°C for 1.5h, shaken to obtain clathrates, and frozen at a low temperature of -10°C 15h, filter and dry to obtain β-cyclodextrin inclusion compound. The obtained clathrate was extracted with petroleum ether, heated to reflux at 90°C, washed with water, and dried. The extract was filtered under reduced p...

Technical field

[0001] The invention belongs to the technical field of biochemical industry, and specifically relates to a reversible reaction-based combined complexation separation method for preparing high-purity unsaturated fatty acids (α-linolenic acid, linoleic acid and oleic acid) from silkworm pupa oil. Background technique

[0002] my country is a big country in the sericulture industry. As a by-product of silk reeling, silkworm pupa can produce nearly 200,000 tons of dried pupa each year, which is an important biomass oil resource. Taking silkworm pupa oil as an example, fat accounts for 9%-10% of fresh silkworm pupa, or 28%-30% of dry matter. Among them, saturated fatty acids account for about 20%, phospholipids and sterols account for about 4%, and unsaturated fatty acids account for about 75%. The unsaturated fatty acid of silkworm pupa oil is mainly composed of α-linolenic acid (about 30%), linoleic acid and oleic acid. It has many physiological functions such as lo...

Examples

Embodiment 1

[0025] The pupa mixed fatty acid and the saturated methanol solution of urea were stirred and mixed at a volume ratio of 1:1, clathrated at 0°C for 24h, the clathrate was cooled and crystallized, filtered, the filtrate obtained was extracted with petroleum ether, heated to reflux at 60°C, and the extract was washed with water , Vacuum degree of 0.001Mpa vacuum filtration, vacuum degree of 0.001Mpa, temperature of 90℃ to recover the solvent to obtain crude unsaturated fatty acid. The crude unsaturated fatty acid was dissolved in ethyl acetate to prepare a saturated solution, and then mixed with saturated β-cyclodextrin aqueous solution at a volume ratio of 1:5, heated in a water bath at 30°C for 0.1h, and shaken to obtain clathrates. Frozen at low temperature for 0.1h, filtered and dried to obtain β-cyclodextrin inclusion compound. The obtained clathrate is extracted with petroleum ether, heated to reflux at 60°C, and the extract is washed with water, and filtered under reduced ...

Embodiment 2

[0027] The mixed fatty acid of silkworm pupa and the saturated n-butanol solution of urea were stirred and mixed in a volume ratio of 1:10, and the inclusion compound was cooled and crystallized at 20°C for 1 h. The obtained filtrate was extracted with n-hexane, heated to reflux at 70°C, and washed with water. The extract is filtered under reduced pressure with a vacuum of 0.098Mpa, a vacuum of 0.098Mpa, and a temperature of 60°C to recover the solvent to obtain crude unsaturated fatty acids. The crude unsaturated fatty acid was dissolved in acetone to prepare a saturated solution, and then mixed with saturated β-cyclodextrin aqueous solution at a volume ratio of 1:15, heated in a water bath at 80°C for 24h, shaken to obtain clathrates, and frozen at 10°C for 24h. Filter and dry to obtain β-cyclodextrin inclusion compound. The obtained clathrate is extracted with petroleum ether, heated to reflux at 90°C, and the extract is washed with water, and the extract is filtered under r...

Embodiment 3

[0029] The pupa mixed fatty acid and the saturated ethanol solution of urea were stirred and mixed at a volume ratio of 1:2, clathrated for 2 hours at 4℃, the clathrate was cooled and crystallized, filtered, the filtrate obtained was extracted with petroleum ether, heated to reflux at 90℃, and the extract was washed with water , Vacuum degree of 0.090Mpa vacuum filtration, vacuum degree of 0.090Mpa, temperature 75℃ to recover the solvent to obtain crude unsaturated fatty acid. The crude unsaturated fatty acid was dissolved in ethanol to prepare a saturated solution, and then mixed with saturated β-cyclodextrin aqueous solution at a volume ratio of 1:9, heated in a water bath at 60°C for 1.5h, shaken to obtain clathrates, and frozen at a low temperature of -10°C 15h, filter and dry to obtain β-cyclodextrin inclusion compound. The obtained clathrate was extracted with petroleum ether, heated to reflux at 90°C, washed with water, and dried. The extract was filtered under reduced p...