CN105153220A - Preparation method of phosphatidylcholine - Google Patents

Abstract

The invention relates to a preparation method of phosphatidylcholine, belonging to the technical field of chemical engineering. The preparation method mainly comprises the following steps: removing oil from rape seed cake, pulverizing into particles, and carrying out microbial fermentation by using an enzyme in Canis familiaris; after the fermentation finishes, submerging the fermentation product by using an ethanol solution, collecting the filtrate, concentrating the filtrate under reduced pressure, taking the supernatant, adding acetone under certain conditions, standing and airing to obtain the phosphatidylcholine. The prepared phosphatidylcholine has higher purity (up to 95% or above). The method has the advantages of lower cost and simple preparation steps.

Description

A kind of preparation method of phosphatidylcholine

Technical field

The present invention relates to a kind of preparation method of phosphatidylcholine, belong to technical field of chemical engineering.

Background technology

Phosphatidylcholine is a kind of natural, safe, efficient emulsifying agent, and can be used for transfusion or used for intravenous injection emulsifying agent, is the most frequently used, the safest emulsifying agent in injection field.Meanwhile, phosphatidylcholine, due to the feature of its liposome, can be used as the carrier of cancer therapy drug and other slow releasing pharmaceuticals, significantly improves the activity of medicine, reduces toxic side effect, have the efficient advantage of low toxicity compared with other medicines carrier.When being used for the purposes such as injection, liposome when phosphatidylcholine, all strict demand is had to the purity of phosphatidylcholine, foreign matter content, water-content, usually require that phosphatidylcholine purity is greater than 95 ~ 98%, the content of phosphatidylethanolamine and Lysophosphatidylcholone is less than 1% simultaneously, water-content lower than 2%, simultaneously to pyrogen, the equal finite quantity requirement of heavy metal content.

The method preparing phosphatidylcholine common in prior art has multiple, wherein has counter-current extraction, and after extraction, purity is relatively high, but full-range yield is still lower, and extraction agent is generally acetonitrile or monohydroxy-alcohol, and extraction selectivity is low, and material loss is serious.Supercritical CO ₂extract the obtained phosphatidylcholine removing neutral fat, purity is only 70%, and ubiquity purity is low, there is the deficiency that yield is low, facility investment is large.The method of separating phospholipids phatidylcholine and membrane sepn combines with column chromatography, the deficiency such as the method exists that film easily pollutes, not easily regeneration, poor repeatability, cost intensive, treatment capacity are little, and separation selectivity is low, can not isolated molecule amount is close completely phospholipid fraction.

Summary of the invention

The technical problem that the present invention mainly solves: for utilizing counter-current extraction, supercritical CO ₂extract and membrane sepn combine with column chromatography partition method obtain phosphatidylcholine, yield is low, purity is low, simultaneously can facility investment large, the problem that required cost is high, provide a kind of preparation method of phosphatidylcholine, first the grease in rapeseed cake is mainly got rid of by the present invention, and be ground into particulate matter, and utilize the enzyme in the stone of a dog's gallbladder, kidney or bladder to carry out fermentable, after having fermented, utilize ethanolic soln to flood fermented product, and collect filtrate, to its concentrating under reduced pressure, get upper liquid, under certain condition, add acetone, leave standstill air-dry.Phosphatidylcholine purity prepared by the present invention is higher, and up to more than 95%, cost is lower simultaneously, and step of preparation process is simple.

In order to solve the problems of the technologies described above, the technical solution adopted in the present invention:

(1) get 1 ~ 2kg rapeseed cake and put into container, get soybean and egg yolk by solid-to-liquid ratio 1:3 to put into juice extractor and squeeze the juice, get the liquid of squeezing out to be placed in container and to soak rapeseed cake, after leaving standstill 20 ~ 30min, the rapeseed cake after soaking is put into pulverizer and is ground into 40 ~ 60 order particulate matter;

(2) particulate matter of above-mentioned gained is put into container, add wherein massfraction be 1 ~ 2% copperas solution flood particulate matter 1 ~ 3cm, at temperature is 80 ~ 90 DEG C, filter after stirring 25 ~ 30min, the filtrate of gained is put into fermentation vat, add 200 ~ 400g stone of a dog's gallbladder, kidney or bladder wherein, after stirring, add 80 ~ 120g glucose respectively wherein again, 300 ~ 400g extractum carnis, 1 ~ 3g dipotassium hydrogen phosphate and 20 ~ 50g ammonium phosphate stir, functional quality mark be 40% sodium hydroxide regulate pH be 6.5 ~ 7.0, sealing and fermenting 10 ~ 15 days, temperature remains 40 ~ 45 DEG C,

(3) after above-mentioned fermentation ends, add wherein massfraction be 80 ~ 85% ethanolic soln flood fermented product 3 ~ 5cm, under rotating speed is 2000 ~ 3000r/min, stir fast filtering after 30 ~ 40min, collect filtered liquid and be placed in container;

(4) said vesse is reduced pressure, be decompressed to 400 ~ 600Pa, and at temperature is 70 ~ 80 DEG C, by liquid concentration to 30 ~ 40% of original volume, by the concentrated solution of gained and propylene glycol by volume 1:1 carry out Homogeneous phase mixing, leave standstill layering after 10 ~ 30min, get upper liquid;

(5) upper liquid of gained is put into container, at temperature is 80 DEG C, after stirring 40 ~ 50min, stir wherein and add the acetone that temperature is 10 ~ 15 DEG C, until produce without precipitation, then filter after leaving standstill 10 ~ 15min, the filtrate of gained is put into air dryer air-dry.

Application of the present invention: be raw material by the phosphatidylcholine of above-mentioned preparation, after adding glycerine or soybean oil dilution, form by gelatin coatings, make lecithin capsule, or directly take in granular form, take this product, effectively can promote brain development, memory, can also remove comedo, skin care simultaneously.

The invention has the beneficial effects as follows:

(1) phosphatidylcholine purity is higher, up to more than 95%;

(2) preparation process is simple, and cost is low.

Embodiment

First get 1 ~ 2kg rapeseed cake and put into container, get soybean and egg yolk by solid-to-liquid ratio 1:3 to put into juice extractor and squeeze the juice, get the liquid of squeezing out to be placed in container and to soak rapeseed cake, after leaving standstill 20 ~ 30min, the rapeseed cake after soaking is put into pulverizer and is ground into 40 ~ 60 order particulate matter, again the particulate matter of above-mentioned gained is put into container, add wherein massfraction be 1 ~ 2% copperas solution flood particulate matter 1 ~ 3cm, at temperature is 80 ~ 90 DEG C, filter after stirring 25 ~ 30min, the filtrate of gained is put into fermentation vat, add 200 ~ 400g stone of a dog's gallbladder, kidney or bladder wherein, after stirring, add 80 ~ 120g glucose respectively wherein again, 300 ~ 400g extractum carnis, 1 ~ 3g dipotassium hydrogen phosphate and 20 ~ 50g ammonium phosphate stir, functional quality mark be 40% sodium hydroxide regulate pH be 6.5 ~ 7.0, sealing and fermenting 10 ~ 15 days, temperature remains 40 ~ 45 DEG C, after above-mentioned fermentation ends, add wherein massfraction be 80 ~ 85% ethanolic soln flood fermented product 3 ~ 5cm, under rotating speed is 2000 ~ 3000r/min, stir fast filtering after 30 ~ 40min, collect filtered liquid and be placed in container, said vesse is reduced pressure, is decompressed to 400 ~ 600Pa, and at temperature is 70 ~ 80 DEG C, by liquid concentration to 30 ~ 40% of original volume, by the concentrated solution of gained and propylene glycol by volume 1:1 carry out Homogeneous phase mixing, leave standstill layering after 10 ~ 30min, get upper liquid, the upper liquid of gained is put into container, at temperature is 80 DEG C, after stirring 40 ~ 50min, stir wherein and add the acetone that temperature is 10 ~ 15 DEG C, until produce without precipitation, then filter after leaving standstill 10 ~ 15min, the filtrate of gained is put into air dryer air-dry.

Example 1

First get 2kg rapeseed cake and put into container, get soybean and egg yolk by solid-to-liquid ratio 1:3 to put into juice extractor and squeeze the juice, get the liquid of squeezing out to be placed in container and to soak rapeseed cake, after leaving standstill 30min, the rapeseed cake after soaking is put into pulverizer and is ground into 60 order particulate matter; Again the particulate matter of above-mentioned gained is put into container, add wherein massfraction be 2% copperas solution flood particulate matter 1cm, at temperature is 90 DEG C, filter after stirring 30min, the filtrate of gained is put into fermentation vat, add the 400g stone of a dog's gallbladder, kidney or bladder wherein, after stirring, add 120g glucose, 400g extractum carnis, 3g dipotassium hydrogen phosphate and 50g ammonium phosphate more wherein respectively to stir, functional quality mark be 40% sodium hydroxide regulate pH be 7.0, sealing and fermenting 15 days, temperature remains 45 DEG C; After above-mentioned fermentation ends, add wherein massfraction be 85% ethanolic soln flood fermented product 5cm, under rotating speed is 3000r/min, stir fast filtering after 40min, collect filtered liquid and be placed in container; Said vesse is reduced pressure, is decompressed to 600Pa, and at temperature is 80 DEG C, by liquid concentration to 40% of original volume, by the concentrated solution of gained and propylene glycol by volume 1:1 carry out Homogeneous phase mixing, leave standstill layering after 30min, get upper liquid; The upper liquid of gained is put into container, at temperature is 80 DEG C, after stirring 50min, stirs wherein and add the acetone that temperature is 15 DEG C, until produce without precipitation, then filter after leaving standstill 15min, the filtrate of gained is put into air dryer air-dry.Be raw material by the phosphatidylcholine of above-mentioned preparation, after adding glycerine or soybean oil dilution, form by gelatin coatings, make lecithin capsule, take 1 every day, each 2, effectively can promote brain development, memory.

Example 2

First get 1kg rapeseed cake and put into container, get soybean and egg yolk by solid-to-liquid ratio 1:3 to put into juice extractor and squeeze the juice, get the liquid of squeezing out to be placed in container and to soak rapeseed cake, after leaving standstill 20min, the rapeseed cake after soaking is put into pulverizer and is ground into 40 order particulate matter; Again the particulate matter of above-mentioned gained is put into container, add wherein massfraction be 1% copperas solution flood particulate matter 1cm, at temperature is 80 DEG C, filter after stirring 25min, the filtrate of gained is put into fermentation vat, add the 200g stone of a dog's gallbladder, kidney or bladder wherein, after stirring, add 80g glucose, 300g extractum carnis, 1g dipotassium hydrogen phosphate and 20g ammonium phosphate more wherein respectively to stir, functional quality mark be 40% sodium hydroxide regulate pH be 6.5, sealing and fermenting 10 days, temperature remains 40 DEG C; After above-mentioned fermentation ends, add wherein massfraction be 80% ethanolic soln flood fermented product 3cm, under rotating speed is 2000r/min, stir fast filtering after 30min, collect filtered liquid and be placed in container; Said vesse is reduced pressure, is decompressed to 400Pa, and at temperature is 70 DEG C, by liquid concentration to 30% of original volume, by the concentrated solution of gained and propylene glycol by volume 1:1 carry out Homogeneous phase mixing, leave standstill layering after 10min, get upper liquid; The upper liquid of gained is put into container, at temperature is 80 DEG C, after stirring 40min, stirs wherein and add the acetone that temperature is 10 DEG C, until produce without precipitation, then filter after leaving standstill 10min, the filtrate of gained is put into air dryer air-dry.Take the particulate matter made with this phosphatidylcholine every day, every day 2, effectively remove comedo on the face, skin care.

Example 3

First get 1kg rapeseed cake and put into container, get soybean and egg yolk by solid-to-liquid ratio 1:3 to put into juice extractor and squeeze the juice, get the liquid of squeezing out to be placed in container and to soak rapeseed cake, after leaving standstill 25min, the rapeseed cake after soaking is put into pulverizer and is ground into 50 order particulate matter; Again the particulate matter of above-mentioned gained is put into container, add wherein massfraction be 1% copperas solution flood particulate matter 2cm, at temperature is 85 DEG C, filter after stirring 27min, the filtrate of gained is put into fermentation vat, add the 300g stone of a dog's gallbladder, kidney or bladder wherein, after stirring, add 100g glucose, 350g extractum carnis, 2g dipotassium hydrogen phosphate and 30g ammonium phosphate more wherein respectively to stir, functional quality mark be 40% sodium hydroxide regulate pH be 6.7, sealing and fermenting 12 days, temperature remains 42 DEG C; After above-mentioned fermentation ends, add wherein massfraction be 83% ethanolic soln flood fermented product 4cm, under rotating speed is 2500r/min, stir fast filtering after 35min, collect filtered liquid and be placed in container; Said vesse is reduced pressure, is decompressed to 500Pa, and at temperature is 75 DEG C, by liquid concentration to 35% of original volume, by the concentrated solution of gained and propylene glycol by volume 1:1 carry out Homogeneous phase mixing, leave standstill layering after 20min, get upper liquid; The upper liquid of gained is put into container, at temperature is 80 DEG C, after stirring 45min, stirs wherein and add the acetone that temperature is 12 DEG C, until produce without precipitation, then filter after leaving standstill 12min, the filtrate of gained is put into air dryer air-dry.Be raw material by the phosphatidylcholine of above-mentioned preparation, after adding glycerine or soybean oil dilution, form by gelatin coatings, make lecithin capsule, take 1 every day, each 1, effectively can promote brain development, memory.

Claims (1)

1. a preparation method for phosphatidylcholine, is characterized in that concrete preparation process is:

(1) get 1 ~ 2kg rapeseed cake and put into container, get soybean and egg yolk by solid-to-liquid ratio 1:3 to put into juice extractor and squeeze the juice, get the liquid of squeezing out to be placed in container and to soak rapeseed cake, after leaving standstill 20 ~ 30min, the rapeseed cake after soaking is put into pulverizer and is ground into 40 ~ 60 order particulate matter;

(2) particulate matter of above-mentioned gained is put into container, add wherein massfraction be 1 ~ 2% copperas solution flood particulate matter 1 ~ 3cm, at temperature is 80 ~ 90 DEG C, filter after stirring 25 ~ 30min, the filtrate of gained is put into fermentation vat, add 200 ~ 400g stone of a dog's gallbladder, kidney or bladder wherein, after stirring, add 80 ~ 120g glucose respectively wherein again, 300 ~ 400g extractum carnis, 1 ~ 3g dipotassium hydrogen phosphate and 20 ~ 50g ammonium phosphate stir, functional quality mark be 40% sodium hydroxide regulate pH be 6.5 ~ 7.0, sealing and fermenting 10 ~ 15 days, temperature remains 40 ~ 45 DEG C,

(3) after above-mentioned fermentation ends, add wherein massfraction be 80 ~ 85% ethanolic soln flood fermented product 3 ~ 5cm, under rotating speed is 2000 ~ 3000r/min, stir fast filtering after 30 ~ 40min, collect filtered liquid and be placed in container;

(4) said vesse is reduced pressure, be decompressed to 400 ~ 600Pa, and at temperature is 70 ~ 80 DEG C, by liquid concentration to 30 ~ 40% of original volume, by the concentrated solution of gained and propylene glycol by volume 1:1 carry out Homogeneous phase mixing, leave standstill layering after 10 ~ 30min, get upper liquid;

(5) upper liquid of gained is put into container, at temperature is 80 DEG C, after stirring 40 ~ 50min, stir wherein and add the acetone that temperature is 10 ~ 15 DEG C, until produce without precipitation, then filter after leaving standstill 10 ~ 15min, the filtrate of gained is put into air dryer air-dry.