CN103864839A - Method for extracting crude phospholipid from cottonseed oil nigre and method for extracting lecithin from crude phospholipid - Google Patents
Method for extracting crude phospholipid from cottonseed oil nigre and method for extracting lecithin from crude phospholipid Download PDFInfo
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Abstract
The invention discloses a method for extracting crude phospholipid from cottonseed oil nigre and a method for extracting lecithin from crude phospholipid. The method for enriching phospholipid from the cottonseed oil nigre, provided by the invention, comprises the following steps: (1), collecting the cottonseed oil nigre, collecting a solid-phase material after oil is removed from acetone, adding petroleum ether, TBHQ (Tert-Butyl Hydroquinone), VE (Vitamin E) and CA (Cellulose Acetate), stirring for 1-5 hours at a temperature of 20-40 DEG C, centrifuging and collecting supernate; (2), collecting the supernate obtained in the step (1), concentrating to obtain a concentrated liquid; and (3), collecting a part by volume of the concentrated liquid obtained in the step (2), adding 1-5 parts by volume of acetone for settling, and collecting a solid-phase material to obtain a target product. According to the method, the problem that the cottonseed oil nigre pollutes the environment is solved, the added value of products is increased, and the rapid development of the cottonseed oil nigre resource comprehensive-utilizing industry is greatly promoted.
Description
Technical field
The method that the present invention relates to extract the method for raw phospholipid from Oleum Gossypii semen soap stock and extract Yelkin TTS from raw phospholipid.
Background technology
China is main Chan Mian big country in the world, and State Statistics Bureau's " national economy and social development statistical communique in 2008 " shows, approximately 5,000,000 hectares of national cotton cultivated areas in 2008,1,000 ten thousand tons of cottonseed ultimate productions.Xinjiang is the province of my domestic cotton maximum, and cotton is the characteristic resources of the tool advantage in Xinjiang, and 2008 Annual planting areas reach 2,000 ten thousand mu, and cottonseed output reaches 4,000,000 tons, accounts for 36% of national ultimate production.Xinjiang cottonseed is large with its output, protein content is high, oil yield is high, favored by domestic processing enterprise.
A cottonseed part is as seed, and all the other all produce grease by traditional cottonseed processing technique (using the method such as screw press, prepressing extraction), and its product is (by sorting out value) such as Oleum Gossypii semen, cottonseed cake (cotton dregs), cotton seed hulls, oil foots (soap stock).
Oleum Gossypii semen soap stock (claiming again cotton oil distillation residues) is the byproduct of Oleum Gossypii semen in alkali refining process, conventionally accounts for the 5-10% of quality of crude oil.Oleum Gossypii semen soap stock is conventionally containing phosphatide 26.3 ± 6.9%.
Phosphatide is the phosphorous fatty substance that a class is present in organic sphere, it is a kind of mixture, mainly (be commonly called as Yelkin TTS by phosphatidylcholine, PC), phosphatidylethanolamine (is commonly called as kephalin, PE), the composition such as phosphatidylserine (PS), phosphatidylinositols (being commonly called as lipositol, PI) and phosphatidic acid (PA).Phosphatide is natural emulsifying agent and tensio-active agent, has higher nutritive value and special physiological property, can be widely used in the fields such as medicine, food, feed, fiber, makeup, leather, dyeing, chemical industry, environmental protection.
In the Oleum Gossypii semen course of processing, if calculate to produce 10% Oleum Gossypii semen soap stock, the annual whole nation is by the Oleum Gossypii semen soap stock that produces 200,000 tons, and resource is very abundant, but most of Oleum Gossypii semen source mill, all using Oleum Gossypii semen soap stock as waste discharge, causes huge pollution and waste.At present, Oleum Gossypii semen soap stock is mainly for the preparation of soap, lipid acid and biofuel, and ignored this high value-added product of phosphatide, caused the significant wastage of phosphatide resource.
Summary of the invention
The object of this invention is to provide the method for extracting the method for raw phospholipid and extract Yelkin TTS from Oleum Gossypii semen soap stock from raw phospholipid.
The method of the first provided by the invention enrichment phosphatide from Oleum Gossypii semen soap stock, comprises the steps:
(1) get Oleum Gossypii semen soap stock, with collecting solid formation after acetone deoiling, add sherwood oil, TBHQ, V
eand CA, 20-40 ℃ is stirred 1-5h, centrifugal and collect supernatant liquor; Described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 20-100ml sherwood oil: 0.002-0.006g TBHQ:0.01-0.03gV
e: 0.002-0.006g CA;
(2) get the supernatant liquor that step (1) obtains, after concentrating, obtain concentrated solution;
(3) get the concentrated solution that 1 parts by volume step (2) obtains, add 1-5 parts by volume acetone to precipitate, collect solid formation, obtain object product.
In described step (1): described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 80ml sherwood oil: 0.002-0.006g TBHQ:0.01-0.03g V
e: 0.002-0.006g CA.In described step (1): described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 80ml sherwood oil: 0.004g TBHQ:0.02g V
e: 0.004g CA.In described step (1): described " 20-40 ℃ is stirred 1-5h " is " 20 ℃ are stirred 2h ".In described step (1): described centrifugal condition specifically can be 3000r/min, 5min.Consider from productive rate, repeatedly repeating step (1), then merges supernatant liquor and carries out step (2).Specifically step (1) can be carried out to twice, then merge supernatant liquor and carry out step (2).
In described step (2): described concentrated method is rotary evaporation.Oleum Gossypii semen soap stock in every 20g step (1) obtains 5 ± 1ml concentrated solution in step (2).
In described step (3): described " 1-5 parts by volume acetone " is " 2 parts by volume acetone ".In described step (3), the method for described " collection solid formation " specifically can be " suction filtration is also collected filter residue ".
Described method also can comprise the steps, the described solid formation that step (3) is obtained is with dry after washing with acetone.
In described step (1), described Oleum Gossypii semen soap stock can carry out repeatedly (as 3 times) acetone deoiling continuously.For 20
goleum Gossypii semen soap stock can add 40mL acetone, then stirring at room temperature 5min at every turn.
The method of the second provided by the invention enrichment phosphatide from Oleum Gossypii semen soap stock, comprises the steps:
(1) get Oleum Gossypii semen soap stock, with collecting solid formation after acetone deoiling, add sherwood oil, TBHQ, V
eand CA, 20-40 ℃ is stirred 1-5h, centrifugal and collect respectively supernatant liquor and precipitation, by supernatant liquor called after supernatant liquor first, will precipitate called after and precipitate first; While preparing described supernatant liquor first, described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 20-100ml sherwood oil: 0.002-0.006g TBHQ:0.01-0.03gV
e: 0.002-0.006g CA;
Get described precipitation first, add sherwood oil, TBHQ, V
eand CA, 20-40 ℃ is stirred 1-5h, centrifugal and collect supernatant liquor, by supernatant liquor called after supernatant liquor second; While preparing described supernatant liquor second, described sherwood oil, described TBHQ, described V
eadd-on while preparing described supernatant liquor first together with the add-on of described CA;
Described supernatant liquor first and described supernatant liquor second are merged, obtain supernatant liquor third;
(2) get the supernatant liquor the third that step (1) obtains, after concentrating, obtain concentrated solution;
(3) get the concentrated solution that 1 parts by volume step (2) obtains, add 1-5 parts by volume acetone to precipitate, collect solid formation, obtain object product.
In described step (1): while preparing described supernatant liquor first, described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 80ml sherwood oil: 0.002-0.006g TBHQ:0.01-0.03g V
e: 0.002-0.006g CA.In described step (1): while preparing described supernatant liquor first, described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 80ml sherwood oil: 0.004g TBHQ:0.02g V
e: 0.004g CA.In described step (1): described centrifugal condition specifically can be 3000r/min, 5min.In described step (1): described " 20-40 ℃ is stirred 1-5h " is " 20 ℃ are stirred 2h ".
In described step (2): described concentrated method is rotary evaporation.Oleum Gossypii semen soap stock in every 20g step (1) obtains 5 ± 1ml concentrated solution in step (2).
In described step (3): described " 1-5 parts by volume acetone " is " 2 parts by volume acetone ".In described step (3), the method for described " collection solid formation " specifically can be " suction filtration is also collected filter residue ".
Described method also can comprise the steps, the described solid formation that step (3) is obtained is with dry after washing with acetone.
In described step (1), described Oleum Gossypii semen soap stock can carry out repeatedly (as 3 times) acetone deoiling continuously.For 20g Oleum Gossypii semen soap stock, can add 40mL acetone, then stirring at room temperature 5min at every turn.
The product (being raw phospholipid, called after product first) that arbitrary described method obtains above also belongs to protection scope of the present invention.
The present invention also protect a kind of from above arbitrary described product first the method for enrichment Yelkin TTS, comprise the steps:
1. get described product first, adding volumn concentration is more than 84% aqueous ethanolic solution or straight alcohol, adds TBHQ, V
eand CA, 15-55 ℃ is stirred 15-75min, then centrifugal collection supernatant liquor;
2. get the supernatant liquor that 1. step obtains, adding discoloring agent and making its concentration is 2-6g/100ml, and 15-55 ℃ is stirred 15-75min, and suction filtration is also collected filtrate;
3. collect the filtrate that 2. step obtains, add ZnCl
2or ZnCl
2the aqueous solution, ZnCl
2quality be the 4-20% of the quality of described product first, stirring reaction 10-50min, suction filtration is also collected filter residue;
4. get the filter residue that 3. step obtains, to colourless, then dry with washing with acetone, obtain dry-matter (product second).
Described step 1. in, described aqueous ethanolic solution is that volumn concentration is 92% aqueous ethanolic solution.Described step 1. in, described " 15-55 ℃ stir 15-75min ", is " 35 ℃ are stirred 45min ".Described step 1. in, described centrifugal parameter is 3000rpm, 5min.
Described step 2. in, the concentration of described discoloring agent is 5g/100ml.Described step 2. in, described " 15-55 ℃ stir 15-75min " be " 30 ℃ of stirring 60min ".Described step 2. in, described discoloring agent is diatomite, aluminium sesquioxide (preferably), bar-shaped gac or granular carbon.
Described step 3. in, ZnCl
2quality be object product claimed in claim 9 quality 12%.Described step 3. in, described " stirring reaction 10-50min " is " stirring reaction 30min ".Described step 3. in, described ZnCl
2the aqueous solution is the ZnCl of 50g/100ml
2the aqueous solution.
Described step 1. in, described product first, described aqueous ethanolic solution, described TBHQ, described V
eas follows with the proportioning of described CA: 4g product first: 16-48ml aqueous ethanolic solution: 0.0006-0.001g TBHQ:0.002-0.006g V
e: 0.0006-0.001g CA.
Described step 1. in, described product first, described aqueous ethanolic solution, described TBHQ, described V
eas follows with the proportioning of described CA: 4g product first: 32ml aqueous ethanolic solution: 0.0008g TBHQ:0.004g V
e: 0.0008g CA.
At present, be mainly oil foot and yolk for the preparation of the raw material of phosphatide, from cotton oil distillation residues, extract phosphatide and do not report both at home and abroad.Adopt method provided by the invention enrichment phosphatide from Oleum Gossypii semen soap stock to have advantages of that extraction yield is high, easy and simple to handle, with low cost.The present invention is for to turn waste into wealth cotton oil distillation residues resource, and the thruster that becomes expansion Cotton industry chain provides new approaches.The problem that the invention solves Oleum Gossypii semen soap stock contaminate environment, has improved added value of product, will carry forward vigorously the fast development of China's cotton oil distillation residues comprehensive utilization of resources industry.
Accompanying drawing explanation
Fig. 1 is the color atlas of Yelkin TTS standard substance.
Fig. 2 is the canonical plotting of Yelkin TTS.
Fig. 3 is the color atlas that detects the lecithin content in the product that the step 11 of embodiment 5 obtains.
Fig. 4 is the result of the step 3 of embodiment 1.
Fig. 5 is the result of the step 4 of embodiment 1.
Fig. 6 is the result of the step 5 of embodiment 1.
Fig. 7 is the result of the step 6 of embodiment 1.
Fig. 8 is the result of the step 7 of embodiment 1.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.Test materials used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples, all arranges and repeats experiment, results averaged for three times.Room temperature in the present embodiment refers to 20 ± 2 ℃.Japan Shimadzu UV-2401PC ultraviolet-visible spectrophotometer.Britain Edwards vacuum freeze drying.The multiplex vacuum pump of Great Wall, Zhengzhou SHB-III circulating water type.Germany Staotious BP211D 100,000/electronic analytical balance.Beijing bright DKZW type electronic thermostatic water-bath, German EYELA Rotary Evaporators.Town in Shanghai booth Anke-40B low speed large capacity centrifuge.The Yingshan Mountains, Hubei SX-2.5-10 type retort furnace.Shimadzu LC-10AT high performance liquid chromatograph.UV-2401 ultraviolet-visible spectrophotometer (Japanese Shimadzu).BP211D electronic analytical balance (German Sai Duolisi).HJ-6B bull temperature control magnetic stirring apparatus (Community of Jin Tan County).Town in Shanghai booth Anke-40B low speed large capacity centrifuge.SHB-III circulating water type vacuum pump (Great Wall, Zhengzhou).Britain Edwards vacuum freeze drying.What the suction filtration in embodiment adopted is qualitative filter paper Xinhua No. 1.
BHT, full name is " 2,6-toluene di-tert-butyl phenol ", is a kind of antioxidant, purchased from Yuan Ye bio tech ltd, Shanghai, article No. is YY12692-500g.BHA, full name is " butylated hydroxy anisole ", is a kind of antioxidant, purchased from Yuan Ye bio tech ltd, Shanghai, article No. is YY13492-250g.TBHQ, full name is " tertiarybutylhydroquinone ", is a kind of antioxidant, purchased from Yuan Ye bio tech ltd, Shanghai, article No. is YY14074-100g.V
ebe vitamin-E, purchased from Yuan Ye bio tech ltd, Shanghai, article No. is YY11518-10g.CA is citric acid, and purchased from Tianjin recovery development in science and technology company limited, article No. is 20111212.Yelkin TTS standard substance, full name is " soy phosphatidylcholine PC ": purchased from sigma company, article No. is P7443.
The measuring method of phospholipid concentration is molybdenum blue colorimetric method (GB/T5537-2008).Typical curve equation is y=5.8218x+0.0132, the quality percentage composition that x is phosphatide, and y is absorbancy, and good in 0.02mg-0.14mg scope internal linear relation, R is 0.9998, and average average recovery is 98.14%-102.37%, and RSD is less than 2%.
Phospholipids content × 100% in phospholipids content ÷ raw material in phosphatide extraction yield=product.
The measuring method of Yelkin TTS is high performance liquid chromatography, Shimadzu LC-10AT high performance liquid chromatograph, chromatographic column be Féraud door Luna silica (2) silicagel column (250 × 4.6mm, 5 μ m); Moving phase: normal hexane: Virahol: water=1:16:4(volume ratio); Detection wavelength is 214nm; Column temperature is 25 ℃; Flow velocity is 0.7mL/min; Sample size is 20 μ l.With Yelkin TTS standard substance production standard curvilinear equation, take Yelkin TTS concentration (μ g/mL) as X-coordinate, peak area (× 10
5) be ordinate zou drawing standard curve, its linear equation is y=0.2018x+1.6163, R is 0.9998, illustrates that Yelkin TTS concentration is good in 28.33～566.67 μ g/mL scope internal linear relations.The color atlas of Yelkin TTS standard substance is shown in Fig. 1, and typical curve is shown in Fig. 2.The color atlas of the product that the step 11 of embodiment 5 obtains is shown in Fig. 3.
Lecithin content × 100% in lecithin content ÷ raw phospholipid in Yelkin TTS extraction yield=product.
Percent of decolourization
The foundation of embodiment 1, method and the optimization of parameter
Oleum Gossypii semen soap stock: Shihezi Kang Long grease Trade Co., Ltd., the quality percentage composition of phosphatide is 23.42%.
The quality percentage composition of phosphatide in the quality × product of the phospholipids content=product in product.
Phospholipids content=100g × 21.05% in raw material.
One, preparation de-oiling soap stock (single part)
(1) get 20g Oleum Gossypii semen soap stock, add 40mL acetone, stirring at room temperature 5min, then suction filtration collect filter residue.
(2) get the filter residue that step (1) obtains, add 40ml acetone, stirring at room temperature 5min, then suction filtration collect filter residue.
(3) get the filter residue that step (2) obtains, add 40ml acetone, stirring at room temperature 5min, then suction filtration collect filter residue, is de-oiling soap stock.
Two, the optimization of anti-oxidant technique
Oxidation index
a
233for sample liquid be 233nm at wavelength absorbancy, A
215for the sample liquid absorbancy that is 215nm at wavelength.
Getting five parts of step 1 obtains de-oiling soap stock (first part is mixed with 80ml sherwood oil; Second part is mixed with BHT and 80ml sherwood oil, and the quality of BHT is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 3rd part is mixed with BHA and 80ml sherwood oil, and the quality of BHA is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 4th part is mixed with TBHQ and 80ml sherwood oil, and the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 5th part and V
eand the mixing of 80ml sherwood oil, V
equality be for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock 0.02%), stirring at room temperature 2 hours, then detects absorbancy.Oxidation index in first part is that the oxidation index in 1.28, the second parts is that oxidation index in 1.03, the three parts is that oxidation index in 1.12, the four parts is that oxidation index in 0.84, the five part is 0.97.The resistance of oxidation of TBHQ is the strongest.
Getting six parts of step 1 obtains de-oiling soap stock (first part is mixed with 80ml sherwood oil; Second part and TBHQ, V
eand the mixing of 80ml sherwood oil, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.01% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 3rd part is mixed with TBHQ, BHT and 80ml sherwood oil, and the quality of TBHQ is for the preparation of 0.01% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, and the quality of BHT is for the preparation of 0.01% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 4th part is mixed with TBHQ, BHA and 80ml sherwood oil, and the quality of TBHQ is for the preparation of 0.01% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, and the quality of BHA is for the preparation of 0.01% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 5th part and TBHQ, V
eand the mixing of 80ml sherwood oil, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.05% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 6th part and TBHQ, V
eand the mixing of 80ml sherwood oil, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock 0.1%), stirring at room temperature 2 hours, then detects absorbancy.Oxidation index in first part is that the oxidation index in 1.33, the second parts is that oxidation index in 0.78, the three part is that oxidation index in 0.95, the four part is that oxidation index in 1.02, the five parts is that oxidation index in 0.72, the six part is 0.67.0.02%TBHQ+0.1%V
eresistance of oxidation the strongest.
Getting six parts of step 1 obtains de-oiling soap stock (first part is mixed with 80ml sherwood oil; Second part is mixed with TBHQ and 80ml sherwood oil, and the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 3rd part is mixed with TBHQ, CA and 80ml sherwood oil, and the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, and the quality of CA is for the preparation of 0.005% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 4th part is mixed with TBHQ, CA and 80ml sherwood oil, and the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, and the quality of CA is for the preparation of 0.01% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 5th part is mixed with TBHQ, CA and 80ml sherwood oil, and the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, and the quality of CA is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 6th part is mixed with TBHQ, CA and 80ml sherwood oil, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA be for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock 0.03%), stirring at room temperature 2 hours, then detects absorbancy.Oxidation index in first part is that the oxidation index in 1.25, the second parts is that oxidation index in 0.85, the three part is that oxidation index in 0.82, the four part is that oxidation index in 0.79, the five part is that oxidation index in 0.77, the six part is 0.96.0.02%TBHQ+0.02%CA is more excellent combination.
Get a step 1 and obtain de-oiling soap stock, with TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be that the quality of CA is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, room temperature leaves standstill 2 hours, then detects absorbancy.Carry out repeating for three times experiment, average oxidation index is 0.61.
Three, the add-on of parameter optimization-sherwood oil
1, get five parts of step 1 and obtain de-oiling soap stock (first part and TBHQ, V
e, CA and 20ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be that the quality of CA is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; Second part and TBHQ, V
e, CA and 40ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be that the quality of CA is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 3rd part and TBHQ, V
e, CA and 60ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be that the quality of CA is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 4th part and TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be that the quality of CA is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock; The 5th part and TBHQ, V
e, CA and 100ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA be for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock 0.02%), stirring at room temperature 2h.
2, the product of every part of step 1 operates respectively as follows: after completing steps 1, centrifugal (3000r/min, 5min), collects supernatant liquor.
3, the product of every part of step 2 operates respectively as follows: get the supernatant liquor that step 2 obtains, be rotated evaporation, obtain the concentrated solution of 5 ± 1ml.
4, the product of every part of step 3 operates respectively as follows: get the concentrated solution that step 3 obtains, add acetone (volume ratio of concentrated solution and acetone is 1:2), suction filtration is also collected filter residue, uses twice of washing with acetone, then lyophilize, the dry-matter obtaining is raw phospholipid (product).
Get respectively each product, detect the quality percentage composition of phosphatide in product and calculate phosphatide extraction yield, the results are shown in Figure 4.Result shows: along with the increase of solid-liquid ratio, the purity of phosphatide declines gradually, and the extraction yield of phosphatide increases gradually, in the time that solid-liquid ratio is greater than 1/4, phospholipid purity and extraction yield change slowly, consider the extraction yield two indexes of phosphatide in phospholipid purity and soap stock, the best proportioning of Oleum Gossypii semen soap stock and sherwood oil is 1g/4ml.
Four, parameter optimization-extraction time
1, get five parts of step 1 and obtain de-oiling soap stock (every portion and TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA be for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock 0.02%), first part of stirring at room temperature 1h, second part of stirring at room temperature 2h, the 3rd part of stirring at room temperature 3h, the 4th part of stirring at room temperature 4h, the 5th part of stirring at room temperature 5h.
2, with 2 of step 3.
3, with 3 of step 3.
4, with 4 of step 3.
Get respectively each product, detect the quality percentage composition of phosphatide in product and calculate phosphatide extraction yield, the results are shown in Figure 5.Result shows: extraction time is little on phospholipid purity and extraction yield impact, and in the time that extraction time is 2h, phosphatide extraction yield is the highest, reaches 54.42%, and prolongation extraction time yield reduces gradually, and its reason may be that partial oxidation has occurred phosphatide.The extraction yield that considers phospholipid purity and phosphatide in raw phospholipid, show that the optimum extraction time is 2h.
Five, parameter optimization-extraction temperature
1, get three parts of step 1 and obtain de-oiling soap stock (every portion and TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA be for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock 0.02%), first part 20 ℃ are stirred 2h, second part 30 ℃ and stir 2h, the 3rd part 40 ℃ and stir 2h.
2, with 2 of step 3.
3, with 3 of step 3.
4, with 4 of step 3.
Get respectively each product, detect the quality percentage composition of phosphatide in product and calculate phosphatide extraction yield, the results are shown in Figure 6.Result shows: in raw phospholipid, the purity of phosphatide declines along with extracting the rising of temperature, the extraction yield of phosphatide is along with the rising of extracting temperature is first risen and declined afterwards, reason is in the time that temperature exceedes 30 ℃, there is partial oxidation in phosphatide, because sherwood oil boiling point is low, in the time of 30 ℃ and 40 ℃, have volatilization in various degree, therefore consider extraction yield and the sherwood oil boiling point of phospholipid purity in raw phospholipid, phosphatide, selecting the suitableeest extraction temperature is 20 ℃.
Six, parameter optimization-acetone add-on
1, get five parts of step 1 and obtain de-oiling soap stock (every portion and TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA be for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock 0.02%), 20 ℃ are stirred 2h.
2, with 2 of step 3.
3, with 3 of step 3.
4, get the concentrated solution that step 3 obtains, (volume ratio of first portion of concentrated solution and acetone is 1:1 to add acetone; The volume ratio of second portion of concentrated solution and acetone is 1:2; The volume ratio of the 3rd portion of concentrated solution and acetone is 1:3; The volume ratio of the 4th portion of concentrated solution and acetone is 1:4; The volume ratio of the 5th portion of concentrated solution and acetone is 1:5), suction filtration is also collected filter residue, and with washing with acetone twice, then lyophilize, the dry-matter obtaining is raw phospholipid (product).
Get respectively each product, detect the quality percentage composition of phosphatide in product and calculate phosphatide extraction yield, the results are shown in Figure 7.Result shows: along with the increase of acetone add-on, phospholipid purity and phosphatide extraction yield are all in slow reduction; When acetone and concentrated solution volume are than 2/1 time, phosphatide has precipitated completely, and phospholipid purity and phosphatide extraction yield are the highest; When acetone adds when excessive, the phosphatide having precipitated has occurred to be again partly dissolved; Although illustrate that phosphatide is insoluble to acetone, excessive acetone still can make phosphatide that a small amount of dissolving occurs.Therefore, the optimal volume proportioning of concentrated solution and acetone is 1:2.
Seven, parameter optimization-extraction time
Get three parts of step 1 and obtain de-oiling soap stock, process respectively as follows:
First part: (1) is by first part of de-oiling soap stock and TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA is that 0.02%, 20 ℃ for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock stirs 2h, centrifugal (3000r/min, 5min), collects supernatant liquor; (2) get the supernatant liquor that step (1) obtains, be rotated evaporation, obtain the concentrated solution of 5 ± 1ml; (3) get the concentrated solution that step (2) obtains, add acetone (volume ratio of concentrated solution and acetone is 1:2), suction filtration is also collected filter residue, and with washing with acetone twice, then lyophilize, the dry-matter obtaining is raw phospholipid (product).
Second part: (1) is by first part of de-oiling soap stock and TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA is that 0.02%, 20 ℃ for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock stirs 2h, centrifugal (3000r/min, 5min), collects respectively supernatant liquor and precipitation; (2) get the precipitation that step (1) obtains, with TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA is that 0.02%, 20 ℃ for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock stirs 2h, centrifugal (3000r/min, 5min), collects supernatant liquor; (3) supernatant liquor that supernatant liquor step (1) being obtained and step (2) obtain merges; (4) get the supernatant liquor that step (3) obtains, be rotated evaporation, obtain the concentrated solution of 5 ± 1ml; (5) get the concentrated solution that step (4) obtains, add acetone (volume ratio of concentrated solution and acetone is 1:2), suction filtration is also collected filter residue, and with washing with acetone twice, then lyophilize, the dry-matter obtaining is raw phospholipid (product).
The 3rd part: (1) is by first part of de-oiling soap stock and TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA is that 0.02%, 20 ℃ for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock stirs 2h, centrifugal (3000r/min, 5min), collects respectively supernatant liquor and precipitation; (2) get the precipitation that step (1) obtains, with TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA is that 0.02%, 20 ℃ for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock stirs 2h, centrifugal (3000r/min, 5min), collects respectively supernatant liquor and precipitation; (3) get the precipitation that step (2) obtains, with TBHQ, V
e, CA and 80ml sherwood oil mix, the quality of TBHQ is for the preparation of 0.02% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, V
equality be for the preparation of 0.1% of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock, the quality of CA is that 0.02%, 20 ℃ for the preparation of the quality of the Oleum Gossypii semen soap stock of de-oiling soap stock stirs 2h, centrifugal (3000r/min, 5min), collects supernatant liquor; (4) supernatant liquor that supernatant liquor step (1) being obtained, the supernatant liquor that step (2) obtains and step (3) obtain merges; (5) get the supernatant liquor that step (4) obtains, be rotated evaporation, obtain the concentrated solution of 5 ± 1ml; (6) get the concentrated solution that step (5) obtains, add acetone (volume ratio of concentrated solution and acetone is 1:2), suction filtration is also collected filter residue, and with washing with acetone twice, then lyophilize, the dry-matter obtaining is raw phospholipid (product).
Get respectively each product, detect the quality percentage composition of phosphatide in product and calculate phosphatide extraction yield, the results are shown in Figure 8.Result shows: along with the increase of extraction time, phosphatide extraction yield increases gradually, purity first increases and then decreases, extracting 2 times compares for 3 times with extracting, its extraction effect is suitable, and purity is the highest while extracting 2 times, consider purity, phosphatide extraction yield and the angle that reduces production costs is set out, selecting optimum extraction time is 2 times.
Eight, orthogonal experiment
On the basis of single factor research of step 3, step 4, step 5, step 6 and step 7, the further impact of 3 factors of volume ratio (C) of mass volume ratio (A), extraction time (B), acetone and the concentrated solution of employing orthogonal test investigation Oleum Gossypii semen soap stock and sherwood oil on phosphatide extraction effect, take phosphatide extraction yield as index, adopt L9 (34) orthogonal table arrangement test, its level of factor coding schedule is in table 1.
Table 1 orthogonal design level of factor table
| Level | A（g/mL） | B（h） | C（v/v） |
| 1 | 1/2 | 1 | 1/1 |
| 2 | 1/3 | 2 | 2/1 |
| 3 | 1/4 | 3 | 3/1 |
Test according to L9 (34) orthogonal table, measure and calculate phosphatide extraction yield, result is analyzed with range method and analysis of variance, refers to table 2 and table 3.
Table 2 orthogonal experiment plan is taken into account interpretation of result
Table 3 analysis of variance table
| Factor | Sum of square of deviations | Degree of freedom | F ratio | F threshold value |
| Solid-liquid ratio | 201.422 | 2 | 5.749 | 19.000 |
| Extraction time | 8.968 | 2 | 0.256 | 19.000 |
| Acetone multiple | 19.958 | 2 | 0.570 | 19.000 |
| Error | 35.03 | 2 | ? | ? |
Note: F0.01(2,2)=99.0, F0.05(2,2)=19.0, F0.10(2,2)=9.0
From range analysis (table 3), each factor on the impact order of phosphatide extraction yield is: A>C>B, i.e. Oleum Gossypii semen soap stock and sherwood oil mass volume ratio > acetone and enriched material volume ratio > extraction time.Be A3B2C2 by the known optimum extraction process of intuitive analysis, the volume ratio that Oleum Gossypii semen soap stock and sherwood oil mass volume ratio are 1/4, extraction time is 2h, acetone and concentrated solution is 2/1, this result with investigate result with single factor and conform to.
Can find out from analysis of variance table (table 3), Oleum Gossypii semen soap stock and sherwood oil mass volume ratio, extraction time, acetone and enriched material volume ratio affect without significance test.Comprehensive experiment of single factor and orthogonal experiments, determine that optimal extract process is: Oleum Gossypii semen soap stock and sherwood oil mass volume ratio are 1/4, extraction temperature is 20 ℃, and extraction time is 2h, and extraction time is 2 times, and acetone and enriched material volume ratio are 2/1.
Embodiment 2, from Oleum Gossypii semen soap stock, extract phosphatide
Oleum Gossypii semen soap stock: Shihezi Kang Long grease Trade Co., Ltd., the quality percentage composition of phosphatide is 21.05%.
1, preparation de-oiling soap stock
(1) get 20g Oleum Gossypii semen soap stock, add 40mL acetone, stirring at room temperature 5min, then suction filtration collect filter residue.
(2) get the filter residue that step (1) obtains, add 40ml acetone, stirring at room temperature 5min, then suction filtration collect filter residue.
(3) get the filter residue that step (2) obtains, add 40ml acetone, stirring at room temperature 5min, then suction filtration collect filter residue, is de-oiling soap stock.
2, get the de-oiling soap stock that step 1 obtains, add 80mL sherwood oil, 0.004g TBHQ, 0.02g V
eand 0.004gCA, 20 ℃ are stirred 2h, and centrifugal (3000r/min, 5min), collects respectively supernatant liquor and precipitation.
3, get the precipitation that step 2 obtains, add 80mL sherwood oil, 0.004g TBHQ, 0.02g V
ewith 0.004g CA, 20 ℃ are stirred 2h, and centrifugal (3000r/min, 5min) collects supernatant liquor.
4, the supernatant liquor that supernatant liquor step 2 being obtained and step 3 obtain merges, and is then rotated evaporation, obtains the concentrated solution of 5 ± 1ml.
5, get the concentrated solution that step 4 obtains, add acetone (volume ratio of concentrated solution and acetone is 1:2), suction filtration is also collected filter residue.
6, get the filter residue that step 5 obtains, with washing with acetone twice, then lyophilize, the dry-matter obtaining is raw phospholipid (product).
7, get the product that step 6 obtains, detect the quality percentage composition of phosphatide wherein and calculate phosphatide extraction yield.
Carry out repeating for three times experiment, the results are shown in Table 4.The average quality percentage composition of phosphatide is that 82.81%, RSD value is less than 1%,
The quality percentage composition of phosphatide and phosphatide extraction yield in table 4 product
| ? | The quality percentage composition (%) of phosphatide in product | Phosphatide extraction yield (%) |
| Repeat for the first time experiment | 83.41 | 66.34 |
| Repeat for the second time experiment | 82.88 | 67.79 |
| Repeat for the third time experiment | 82.16 | 66.85 |
| On average | 82.81 | 66.98 |
| RSD(%) | 0.77 | 1.09 |
Embodiment 3, from Oleum Gossypii semen soap stock, extract phosphatide
Oleum Gossypii semen soap stock: Shihezi Kang Long grease Trade Co., Ltd., the quality percentage composition of phosphatide is 19.37%.
Method with reference to embodiment 2 operates that (difference is only step 2 and step 3), the results are shown in Table 5.
2, get the de-oiling soap stock that step 1 obtains, add 80mL sherwood oil, 0.002g TBHQ, 0.01g V
eand 0.002gCA, 20 ℃ are stirred 2h, and centrifugal (3000r/min, 5min), collects respectively supernatant liquor and precipitation.
3, get the precipitation that step 2 obtains, add 80mL sherwood oil, 0.002g TBHQ, 0.01g V
ewith 0.002g CA, 20 ℃ are stirred 2h, and centrifugal (3000r/min, 5min) collects supernatant liquor.
The quality percentage composition of phosphatide and phosphatide extraction yield in table 5 product
| ? | The quality percentage composition (%) of phosphatide in product | Phosphatide extraction yield (%) |
| Repeat for the first time experiment | 48.62 | 77.14 |
| Repeat for the second time experiment | 47.05 | 79.32 |
| Repeat for the third time experiment | 49.47 | 78.47 |
| On average | 48.38 | 78.31 |
| RSD(%) | 1.23 | 1.41 |
Embodiment 4, from Oleum Gossypii semen soap stock, extract phosphatide
Oleum Gossypii semen soap stock: Shihezi Kang Long grease Trade Co., Ltd., the quality percentage composition of phosphatide is 21.05%.
Method with reference to embodiment 2 operates that (difference is only step 2 and step 3), the results are shown in Table 6.
2, get the de-oiling soap stock that step 1 obtains, add 80mL sherwood oil, 0.006g TBHQ, 0.03g V
eand 0.006gCA, 20 ℃ are stirred 2h, and centrifugal (3000r/min, 5min), collects respectively supernatant liquor and precipitation.
3, get the precipitation that step 2 obtains, add 80mL sherwood oil, 0.006g TBHQ, 0.03g V
ewith 0.006g CA, 20 ℃ are stirred 2h, and centrifugal (3000r/min, 5min) collects supernatant liquor.
The quality percentage composition of phosphatide and phosphatide extraction yield in table 6 product
| ? | The quality percentage composition (%) of phosphatide in product | Phosphatide extraction yield (%) |
| Repeat for the first time experiment | 53.14 | 81.33 |
| Repeat for the second time experiment | 51.87 | 79.98 |
| Repeat for the third time experiment | 51.44 | 79.26 |
| On average | 52.15 | 80.19 |
| RSD(%) | 1.69 | 1.31 |
Embodiment 5, from raw phospholipid, extract Yelkin TTS
One, parameter optimization-alcohol concn
1, get and in six parts of embodiment 2, repeat for the first time raw phospholipid (every part of 4.0g raw phospholipid that experiment prepares; First part adds 32mL100% ethanol, and second part adds 32mL84% aqueous ethanolic solution, and the 3rd part adds 32mL88% aqueous ethanolic solution, and the 4th part adds 32mL92% aqueous ethanolic solution, and the 5th part adds 32mL96% aqueous ethanolic solution; % in this bracket all represents volume ratio), add TBHQ, V
eand CA, 0.02% of the quality that the quality of TBHQ is raw phospholipid, V
ethe quality quality that is raw phospholipid 0.1%, 0.02%, 35 ℃ of the quality that the quality of CA is raw phospholipid is stirred 45min, the then centrifugal 5min of 3000rpm, collects supernatant liquor.
2, the product of every part of step 1 operates respectively as follows: get the supernatant liquor that step 1 obtains, adding discoloring agent (aluminium sesquioxide) and making its concentration is 5g/100ml, and 30 ℃ are stirred 60min, and suction filtration is also collected filtrate.
3, the product of every part of step 2 operates respectively as follows: collect the filtrate that step 2 obtains, add precipitation agent (50g/100mlZnCl
2the aqueous solution), ZnCl
2the quality quality that is raw phospholipid 12%, stirring reaction 30min, suction filtration is also collected filter residue.
4, the product of every part of step 3 operates respectively as follows: get the filter residue that step 3 obtains, with 5 ℃ of washing with acetones, to colourless, then lyophilize, obtains dry-matter (product).
The purity of lecithin of first part of product is 64.37%.
The purity of lecithin of second part of product is 53.68%.
The purity of lecithin of the 3rd part of product is 73.03%.
The purity of lecithin of the 4th part of product is 82.25%.
The purity of lecithin of the 5th part of product is 75.19%.
The purity of lecithin of the 4th part of product is significantly higher than other product, is optimized parameter so select 92% aqueous ethanolic solution.
Two, parameter optimization-ethanol volume
1, get and in five parts of embodiment 2, repeat for the first time raw phospholipid (every part of 4.0g raw phospholipid that experiment prepares; First part adds 16mL92% aqueous ethanolic solution, and second part adds 24mL92% aqueous ethanolic solution, and the 3rd part adds 32mL92% aqueous ethanolic solution, the 4th part adds 40mL92% aqueous ethanolic solution, the 5th part adds 48mL92% aqueous ethanolic solution, and the % in this bracket all represents volume ratio), add TBHQ, V
eand CA, 0.02% of the quality that the quality of TBHQ is raw phospholipid, V
ethe quality quality that is raw phospholipid 0.1%, 0.02%, 35 ℃ of the quality that the quality of CA is raw phospholipid is stirred 45min, the then centrifugal 5min of 3000rpm, collects supernatant liquor.
2, with 2 of step 1.
3, with 3 of step 1.
4, with 4 of step 1.
The purity of lecithin of first part of product is 34.17%.
The purity of lecithin of second part of product is 56.03%.
The purity of lecithin of the 3rd part of product is 74.32%.
The purity of lecithin of the 4th part of product is 79.17%.
The purity of lecithin of the 5th part of product is 83.43%.
The purity of lecithin of the 3rd part, the 4th part and the 5th part product is all higher, considers based on cost angle, and selecting 32mL aqueous ethanolic solution is optimized parameter.
Three, parameter optimization-extraction temperature
1, get and in five parts of embodiment 2, repeat for the first time (every part 4.0 of the raw phospholipid that experiment prepares
graw phospholipid; Every portion adds 32mL92% aqueous ethanolic solution; % in this bracket all represents volume ratio), add TBHQ, V
eand CA, 0.02% of the quality that the quality of TBHQ is raw phospholipid, V
ethe quality quality that is raw phospholipid 0.1%, the quality of CA be raw phospholipid quality 0.02%, first part 15 ℃ are stirred 45min, second part 25 ℃ stirring 45min, the 3rd part 35 ℃ stirring 45min, the 4th part 45 ℃ stirring 45min, the 5th part 55 ℃ stirring 45min, then the centrifugal 5min of 3000rpm, collects supernatant liquor.
2, with 2 of step 1.
3, with 3 of step 1.
4, with 4 of step 1.
The purity of lecithin of first part of product is 61.75%.
The purity of lecithin of second part of product is 83.96%.
The purity of lecithin of the 3rd part of product is 87.27%.
The purity of lecithin of the 4th part of product is 73.43%.
The purity of lecithin of the 5th part of product is 41.11%.
The purity of lecithin of the 3rd part of product is significantly higher than other product, is optimized parameter so select 35 ℃.
Four, parameter optimization-extraction time
1, get and in five parts of embodiment 2, repeat for the first time (every part 4.0 of the raw phospholipid that experiment prepares
graw phospholipid; Every portion adds 32mL92% aqueous ethanolic solution; % in this bracket all represents volume ratio), add TBHQ, V
eand CA, 0.02% of the quality that the quality of TBHQ is raw phospholipid, V
ethe quality quality that is raw phospholipid 0.1%, the quality of CA be raw phospholipid quality 0.02%, first part 35 ℃ are stirred 15min, second part 35 ℃ stirring 30min, the 3rd part 35 ℃ stirring 45min, the 4th part 35 ℃ stirring 60min, the 5th part 35 ℃ stirring 75min, then the centrifugal 5min of 3000rpm, collects supernatant liquor.
2, with 2 of step 1.
3, with 3 of step 1.
4, with 4 of step 1.
The purity of lecithin of first part of product is 64.42%.
The purity of lecithin of second part of product is 81.35%.
The purity of lecithin of the 3rd part of product is 85.42%.
The purity of lecithin of the 4th part of product is 86.21%.
The purity of lecithin of the 5th part of product is 85.03%.
The purity of lecithin of the 3rd part, the 4th part and the 5th part product is all higher, considers based on cost angle, and selection 45min is optimized parameter.
Five, parameter optimization-discoloring agent
1, get in four parts of embodiment 2 and repeat for the first time the raw phospholipid (every part of 4.0g) that experiment prepares, add 32mL92%(volume ratio) aqueous ethanolic solution, add TBHQ, V
eand CA, 0.02% of the quality that the quality of TBHQ is raw phospholipid, V
ethe quality quality that is raw phospholipid 0.1%, 0.02%, 35 ℃ of the quality that the quality of CA is raw phospholipid is stirred 45min, the then centrifugal 5min of 3000rpm, collects supernatant liquor.
2, get the supernatant liquor that step 1 obtains, (first part of employing is discoloring agent as diatomite to add discoloring agent, second part adopts aluminium sesquioxide as discoloring agent, the 3rd part adopts bar-shaped gac as discoloring agent, the 4th part adopts granular carbon as discoloring agent) and to make its concentration be 5g/100ml, 30 ℃ are stirred 60min, and suction filtration is also collected filtrate.
The absorbancy (A1) of the supernatant liquor that 3, under 383nm wavelength, detecting step 1 obtains respectively and the absorbancy (A2) of the filtrate that step 2 obtains, calculate percent of decolourization.
The percent of decolourization of first part is 18.14%.
The percent of decolourization of second part is 58.69%.
The percent of decolourization of the 3rd part is 2.43%.
The percent of decolourization of the 4th part is 49.02%.
The percent of decolourization of second part is the highest, so selection aluminium sesquioxide is optimized parameter.
Six, parameter optimization-discoloring agent concentration
1, get in five parts of embodiment 2 and repeat for the first time the raw phospholipid (every part of 4.0g) that experiment prepares, add 32mL92%(volume ratio) aqueous ethanolic solution, add TBHQ, V
eand CA, 0.02% of the quality that the quality of TBHQ is raw phospholipid, V
ethe quality quality that is raw phospholipid 0.1%, 0.02%, 35 ℃ of the quality that the quality of CA is raw phospholipid is stirred 45min, the then centrifugal 5min of 3000rpm, collects supernatant liquor.
2, get the supernatant liquor that step 1 obtains, add discoloring agent (aluminium sesquioxide) and make its concentration for (first part for 2g/100ml, second part for 3g/100ml, the 3rd part for 4g/100ml, the 4th part for 5g/100ml, the 5th part be 6g/100ml), 30 ℃ are stirred 60min, and suction filtration is also collected filtrate.
The absorbancy (A1) of the supernatant liquor that 3, under 383nm wavelength, detecting step 1 obtains respectively and the absorbancy (A2) of the filtrate that step 2 obtains, calculate percent of decolourization.
The percent of decolourization of first part is 42.35%.
The percent of decolourization of second part is 55.70%.
The percent of decolourization of the 3rd part of thing is 78.14%.
The percent of decolourization of the 4th part is 86.56%.
The percent of decolourization of the 5th part is 87.02%.
The percent of decolourization of the 4th part and the 5th part is all higher, considers based on cost angle, and selection 5g/100ml is optimized parameter.
Seven, parameter optimization-bleaching time
1, with 1 of step 6.
2, get the supernatant liquor that step 1 obtains, adding discoloring agent (aluminium sesquioxide) and making its concentration is 5g/100ml, first part 30 ℃ stir 15min, second part 30 ℃ stir 30min, the 3rd part 30 ℃ stir 45min, the 4th part 30 ℃ stir 60min, the 5th part 30 ℃ stir 75min,, suction filtration is also collected filtrate.
The absorbancy (A1) of the supernatant liquor that 3, under 383nm wavelength, detecting step 1 obtains respectively and the absorbancy (A2) of the filtrate that step 2 obtains, calculate percent of decolourization.
The percent of decolourization of first part is 68.93%.
The percent of decolourization of second part is 73.31%.
The percent of decolourization of the 3rd part is 81.43%.
The percent of decolourization of the 4th part is 88.68%.
The percent of decolourization of the 5th part is 85.12%.
The percent of decolourization of the 4th part is the highest, so selection 60min is optimized parameter.
Eight, parameter optimization-bleaching temperature
1, with 1 of step 6.
2, get the supernatant liquor that step 1 obtains, adding discoloring agent (aluminium sesquioxide) and making its concentration is 5g/100ml, first part 15 ℃ stir 60min, second part 25 ℃ stir 60min, the 3rd part 30 ℃ stir 60min, the 4th part 45 ℃ stir 60min, the 5th part 55 ℃ stir 60min,, suction filtration is also collected filtrate.
The absorbancy (A1) of the supernatant liquor that 3, under 383nm wavelength, detecting step 1 obtains respectively and the absorbancy (A2) of the filtrate that step 2 obtains, calculate percent of decolourization.
The percent of decolourization of first part is 71.48%.
The percent of decolourization of second part is 82.68%.
The percent of decolourization of the 3rd part is 90.08%.
The percent of decolourization of the 4th part is 91.44%.
The percent of decolourization of the 5th part is 92.84%.
The percent of decolourization of the 3rd part, the 4th part and the 5th part is all higher, considers based on cost angle, and selecting 30 ℃ is optimized parameter.
Nine, parameter optimization-precipitation agent add-on
1, with 1 of step 6.
2, with 2 of step 1.
3, collect the filtrate that step 2 obtains, add precipitation agent (50g/100ml ZnCl
2the aqueous solution), ZnCl in first part
2the quality quality that is raw phospholipid 4%, ZnCl in second part
2the quality quality that is raw phospholipid 8%, ZnCl in the 3rd part
2the quality quality that is raw phospholipid 12%, ZnCl in the 4th part
2the quality quality that is raw phospholipid 16%, ZnCl in the 5th part
2the quality quality that is raw phospholipid 20%, stirring reaction 30min, suction filtration is also collected filter residue.
4, with 4 of step 1.
The purity of lecithin of first part of product is 76.17%, and yield is 15.75%.
The purity of lecithin of second part of product is 79.35%, and yield is 22.25%.
The purity of lecithin of the 3rd part of product is 82.41%, and yield is 23.84%.
The purity of lecithin of the 4th part of product is 71.24%, and yield is 27.27%.
The purity of lecithin of the 5th part of product is 64.51%, and yield is 30.68%.
The purity of lecithin of the 3rd part of product is significantly higher than other product, so select 12% to be optimized parameter.
Ten, parameter optimization-sedimentation time
1, with 1 of step 6.
2, with 2 of step 1.
3, collect the filtrate that step 2 obtains, add precipitation agent (50g/100ml ZnCl
2the aqueous solution), ZnCl
212%, the first part of stirring reaction 10min, second part of stirring reaction 20min, the 3rd part of stirring reaction 30min, the 4th part of stirring reaction 40min, the 5th part of stirring reaction 50min of the quality quality that is raw phospholipid, suction filtration is also collected filter residue.
4, with 4 of step 1.
The purity of lecithin of first part of product is 79.15%, and yield is 16.73%.
The purity of lecithin of second part of product is 81.49%, and yield is 24.17%.
The purity of lecithin of the 3rd part of product is 82.16%, and yield is 24.48%.
The purity of lecithin of the 4th part of product is 80.57%, and yield is 23.64%.
The purity of lecithin of the 5th part of product is 81.29%, and yield is 23.17%.
The purity of lecithin of the 3rd part of product is significantly higher than other product, so selection 30min is optimized parameter.
11, adopt all parameters after optimizing from raw phospholipid, to extract Yelkin TTS
1, get in 4.0g embodiment 2 and repeat for the first time the raw phospholipid that experiment prepares, add 32mL92%(volume ratio) aqueous ethanolic solution, add TBHQ, V
eand CA, 0.02% of the quality that the quality of TBHQ is raw phospholipid, V
ethe quality quality that is raw phospholipid 0.1%, 0.02%, 35 ℃ of the quality that the quality of CA is raw phospholipid is stirred 45min, the then centrifugal 5min of 3000rpm, collects supernatant liquor.
2, get the supernatant liquor that step 1 obtains, adding discoloring agent (aluminium sesquioxide) and making its concentration is 5g/100ml, and 30 ℃ are stirred 60min, and suction filtration is also collected filtrate.
3, collect the filtrate that step 2 obtains, add the precipitation agent (ZnCl of 50g/100ml
2the aqueous solution), ZnCl
2the quality quality that is raw phospholipid 12%, stirring reaction 30min, suction filtration is also collected filter residue.
4, get the filter residue that step 3 obtains, with 5 ℃ of washing with acetones, to colourless, then lyophilize, obtains dry-matter (product).
In product, total phospholipids purity is 89.51%.In product, purity of lecithin is 82.41%, and yield is 23.31%.
Claims (10)
1. a method for enrichment phosphatide from Oleum Gossypii semen soap stock, comprises the steps:
(1) get Oleum Gossypii semen soap stock, with collecting solid formation after acetone deoiling, add sherwood oil, TBHQ, V
eand CA, 20-40 ℃ is stirred 1-5h, centrifugal and collect supernatant liquor; Described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 20-100ml sherwood oil: 0.002-0.006g TBHQ:0.01-0.03gV
e: 0.002-0.006g CA;
(2) get the supernatant liquor that step (1) obtains, after concentrating, obtain concentrated solution;
(3) get the concentrated solution that 1 parts by volume step (2) obtains, add 1-5 parts by volume acetone to precipitate, collect solid formation, obtain object product.
2. the method for claim 1, is characterized in that: in described step (1):
Described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 80ml sherwood oil: 0.002-0.006g TBHQ:0.01-0.03g V
e: 0.002-0.006g CA;
Described " 20-40 ℃ is stirred 1-5h " is " 20 ℃ are stirred 2h ".
3. method as claimed in claim 2, is characterized in that: in described step (1):
Described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 80ml sherwood oil: 0.004g TBHQ:0.02g V
e: 0.004g CA.
4. a method for enrichment phosphatide from Oleum Gossypii semen soap stock, comprises the steps:
(1) get Oleum Gossypii semen soap stock, with collecting solid formation after acetone deoiling, add sherwood oil, TBHQ, V
eand CA, 20-40 ℃ is stirred 1-5h, centrifugal and collect respectively supernatant liquor and precipitation, by supernatant liquor called after supernatant liquor first, will precipitate called after and precipitate first; While preparing described supernatant liquor first, described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 20-100ml sherwood oil: 0.002-0.006g TBHQ:0.01-0.03gV
e: 0.002-0.006g CA;
Get described precipitation first, add sherwood oil, TBHQ, V
eand CA, 20-40 ℃ is stirred 1-5h, centrifugal and collect supernatant liquor, by supernatant liquor called after supernatant liquor second; While preparing described supernatant liquor second, described sherwood oil, described TBHQ, described V
eadd-on while preparing described supernatant liquor first together with the add-on of described CA;
Described supernatant liquor first and described supernatant liquor second are merged, obtain supernatant liquor third;
(2) get the supernatant liquor the third that step (1) obtains, after concentrating, obtain concentrated solution;
(3) get the concentrated solution that 1 parts by volume step (2) obtains, add 1-5 parts by volume acetone to precipitate, collect solid formation, obtain object product.
5. method as claimed in claim 4, is characterized in that: in described step (1):
While preparing described supernatant liquor first, described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 80ml sherwood oil: 0.002-0.006g TBHQ:0.01-0.03g V
e: 0.002-0.006g CA;
Described " 20-40 ℃ is stirred 1-5h " is " 20 ℃ are stirred 2h ".
6. method as claimed in claim 5, is characterized in that: in described step (1):
While preparing described supernatant liquor first, described Oleum Gossypii semen soap stock, described sherwood oil, described TBHQ, described V
eas follows with the proportioning of described CA: 20g Oleum Gossypii semen soap stock: 80ml sherwood oil: 0.004g TBHQ:0.02g V
e: 0.004g CA.
7. as the method as described in arbitrary in claim 1 to 6, it is characterized in that: in described step (2):
Described concentrated method is rotary evaporation;
Oleum Gossypii semen soap stock in every 20g step (1) obtains 5 ± 1ml concentrated solution in step (2).
8. as the method as described in arbitrary in claim 1 to 7, it is characterized in that: in described step (3):
Described " 1-5 parts by volume acetone " is " 2 parts by volume acetone ".
9. the object product that in claim 1 to 8, arbitrary described method obtains.
10. Accessory Right requires the method for enrichment Yelkin TTS in the object product described in 9, comprises the steps:
1. get the object product described in claim 9, adding volumn concentration is more than 84% aqueous ethanolic solution or straight alcohol, adds TBHQ, V
eand CA, 15-55 ℃ is stirred 15-75min, then centrifugal collection supernatant liquor;
2. get the supernatant liquor that 1. step obtains, adding discoloring agent and making its concentration is 2-6g/100ml, and 15-55 ℃ is stirred 15-75min, and suction filtration is also collected filtrate;
3. collect the filtrate that 2. step obtains, add ZnCl
2or ZnCl
2the aqueous solution, ZnCl
2quality be the 4-20% of the quality of object product claimed in claim 9, stirring reaction 10-50min, suction filtration is also collected filter residue;
4. get the filter residue that 3. step obtains, to colourless, then dry with washing with acetone, obtain dry-matter.
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| CN201410068133.2A CN103864839B (en) | 2014-02-26 | 2014-02-26 | From Oleum Gossypii semen soap stock, extract the method for raw phospholipid and from raw phospholipid, extract the method for Yelkin TTS |
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| CN201410068133.2A CN103864839B (en) | 2014-02-26 | 2014-02-26 | From Oleum Gossypii semen soap stock, extract the method for raw phospholipid and from raw phospholipid, extract the method for Yelkin TTS |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104397322A (en) * | 2014-11-06 | 2015-03-11 | 江南大学 | Preparation method of oriental sesame phospholipid |
| CN105494887A (en) * | 2015-12-19 | 2016-04-20 | 仇颖超 | Method for extracting soya bean lecithin from soybean curb residues |
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|---|---|---|---|---|
| US6426423B1 (en) * | 1998-11-20 | 2002-07-30 | I.P. Holdings | Methods for treating phosphatide-containing mixtures |
| US20030050492A1 (en) * | 1998-11-20 | 2003-03-13 | Ip Holdings, L.L.C. | Vegetable oil refining |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6426423B1 (en) * | 1998-11-20 | 2002-07-30 | I.P. Holdings | Methods for treating phosphatide-containing mixtures |
| US20030050492A1 (en) * | 1998-11-20 | 2003-03-13 | Ip Holdings, L.L.C. | Vegetable oil refining |
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| Title |
|---|
| 王岚: "棉籽油皂脚的开发与利用", 《中国油脂》, vol. 30, no. 2, 31 December 2005 (2005-12-31), pages 34 - 36 * |
| 赵洪涛等: "棉油皂脚中磷脂的提取工艺研究", 《食品科技》, vol. 38, no. 4, 31 December 2013 (2013-12-31), pages 256 - 260 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104397322A (en) * | 2014-11-06 | 2015-03-11 | 江南大学 | Preparation method of oriental sesame phospholipid |
| CN105494887A (en) * | 2015-12-19 | 2016-04-20 | 仇颖超 | Method for extracting soya bean lecithin from soybean curb residues |
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| CN103864839B (en) | 2016-01-06 |
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