CN105067749A - Extraction and detection method of defatted Euphausia superba erucic acid - Google Patents
Extraction and detection method of defatted Euphausia superba erucic acid Download PDFInfo
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Abstract
The present invention discloses an extraction and detection method of defatted Euphausia superba erucic acid. The method comprises: (1) carrying out heating reflux extraction on defatted Euphausia superba with methanol, and removing methanol to obtain a paste; (2) taking a chromatography column, drying silica gel, wetting with petroleum ether, and packing; (3) loading the paste in the step (1) into the column, carrying out gradient elution, and collecting the eluents in a section manner, wherein eluants are different proportions of petroleum ether and ethyl acetate, and the elution is performed according to the polarity from the low polarity eluant to the high polarity eluant; (4) carrying out primary qualitation on the collected eluents by using a thin layer chromatography method, merging and expanding, simply dotting the eluent with the Rf of 0.5, evaporating, and volatilizing to obtain an erucic acid sample; and (5) carrying out intermediate infrared transmittance detection. The present invention provides the extraction and detection of the defatted Euphausia superba erucic acid, wherein the detection method is simple, rapid, accurate, stable and reliable, and the yield of the extraction method can achieve 2.19%.
Description
Technical field
The present invention relates to a kind of isolation and determination method of degreasing krill erucic acid, belong to food, chemistry and field of medicaments.
Background technology
Erucic acid has another name called cis-13-22 carbon monoenoic acid, is a kind of long-chain unsaturated fatty acid, is mainly present in the vegetable seeds of Cruciferae and trollflower section, also find that there is the existence of erucic acid in some marine animal greases.One of purposes of erucic acid produces erucyl amide.Can also for the manufacture of regenerated fiber, polyester and textile auxiliary, PVC stabilizer, the agent of paint dryness, surface-coating resins and processing behenic acid etc.Erucic acid and glyceride thereof can be applicable to food industry or cosmetic formulation art; For the production of surfactant (washing agent), lubricant, also for Biochemical Research, organic synthesis etc.Erucic acid is a kind of very important oil-fat chemical products, has extensive use in metallurgy, machinery, rubber, chemical industry, paint, weaving and medicine and other fields.Erucic acid is used for aluminum paste, good polishing action can be played, make it in color and luster, tack etc., all show good performance.Erucic acid is of many uses with it, added value is high, the huge market demand and the feature such as renewable, is just demonstrating more and more important effect, be described as the important fine chemical material of 21 century by expert at modern times many industrial circles.
Krill (EuphausiasuperbaDana) is one of single living resources maximum on the earth, and its standing crop is estimated as 6.5-10 hundred million tons, year amount of fishing can reach 100,000,000 tons.This is equivalent to the whole world fish of a year and crustaceanly fishes for summation, and nervous for alleviation world food, shortage of resources, has very important meaning.Krill is also a kind of huge living resources newly preparing erucic acid.
Summary of the invention
The object of this invention is to provide a kind of quick, easy, accurate, reliable krill erucic acid isolation and determination method.
The technical solution used in the present invention is for achieving the above object:
An extracting method for degreasing krill erucic acid, comprises the following steps:
(1) degreasing krill is adopted methyl alcohol heating and refluxing extraction, removing methyl alcohol, obtains paste;
(2) chromatographic column is got, with sherwood oil wetting dress post after silica dehydrator;
(3) the paste loading in step (1), gradient elution, eluant, eluent is sherwood oil and the ethyl acetate of different proportion, according to polarity wash-out from low to high, and Fractional Collections eluent;
(4) collected eluent is adopted thin-layered chromatography initial characterization, merge after launching and only have single point and R
fthe eluent of=0.50, evaporation, volatilizes to obtain erucic acid sample.
A detection method for degreasing krill erucic acid, comprises the following steps:
(1) degreasing krill is adopted methyl alcohol heating and refluxing extraction, removing methyl alcohol, obtains paste;
(2) chromatographic column is got, with sherwood oil wetting dress post after silica dehydrator;
(3) the paste loading in step (1), carries out gradient elution by low polarity to high polarity, and eluant, eluent is sherwood oil and the ethyl acetate of different proportion, Fractional Collections eluent;
(4) collected eluent is adopted thin-layered chromatography initial characterization, merge after launching and only have single point and R
fthe eluent of=0.50, evaporation, volatilizes to obtain erucic acid sample.
(5) adopt KBr pressed disc method to carry out compressing tablet in the erucic acid sample in step (4), gained compressing tablet carries out the detection of middle infrared light transmission rate (MIR-TR).
In step (1), described degreasing krill is for adopting hydrophobic solvent by krill except degrease, and described hydrophobic solvent is normal hexane, vegetable oil extractant or sherwood oil.
In step (1), the mass volume ratio of described degreasing krill and methyl alcohol is 1g:(6 ~ 10) mL, preferred mass volume ratio is 1g:6mL, adopt methyl alcohol heating and refluxing extraction 2 ~ 5 times, be preferably 3 times, Extracting temperature is 70 ~ 80 DEG C, and extraction time is 60min.
Prove through a large amount of experiments, when the mass volume ratio of degreasing krill and methyl alcohol is 1g:(6 ~ 10) mL time, the extraction effect of erucic acid sample is best.
In step (1), the water cut of described degreasing krill is 7 ~ 8%.
In step (2), described chromatographic column specification is 3 × 30cm
2, used silica gel specification is 200-300 order.The silicagel column of different size is investigated, finds to use the separating effect of 200-300 order silica gel best.
In step (2), silica gel fills post with sherwood oil is wetting after dry 8 ~ 12h at 100 ~ 110 DEG C.
In step (3), the volume ratio of described sherwood oil and ethyl acetate is 1:0, (50 ~ 0): 1, and the elution volume of each gradient is 500mL; When sherwood oil: when the volume ratio of ethyl acetate is 1:0, the polarity of eluant, eluent is minimum, when sherwood oil: when the volume of ethyl acetate is 0:1, and the polarity of eluant, eluent is maximum.
One preferably embodiment is: in step (3), the eluent petroleum ether of opposed polarity gradient and the volume ratio of ethyl acetate are respectively: 0:1,50:1,30:1,20:1,15:1,10:1,3:1,1:1,1:0, and the elution volume of each gradient is 500mL.
In step (4), the thin-layer developing plate that thin-layered chromatography uses is GF
254high Performance Thin plate (10 × 10cm2), developping agent is dimethylbenzene: ethyl acetate: glacial acetic acid=9:2:0.5, and exhibition distance is 8cm.Qualitative coloring agent used is massfraction is 5 ~ 15% sulfuric acid solutions, and being preferably massfraction is the sulfuric acid solution of 10%, dries 5min for 110 DEG C.
After developping agent of the present invention launches, component spot circle, clear, launches fast.
Different developer is investigated, selects massfraction to be 10%H
2sO
4when solution is as developer, develop the color sensitive, effect is best.
In step (5), tabletting conditions: the mass ratio of sample and KBr is 1:4, pressure 12MPa, duration 30s.MIR-TR testing conditions: resolution 4cm
-1, sweep time 16s, mid-infrared light source, KBr beam splitter.
The invention has the beneficial effects as follows:
(1) the invention provides extraction and the detection method of krill erucic acid, detection method is simple, fast, accurately, stable, reliably, the yield of extracting method can reach 2.19%.
(2) the present invention from abundant, extract erucic acid renewable resource degreasing krill, develop a kind of new resources preparing erucic acid.
(3) the present invention makes the value of krill obtain further development and utilization, and thinking has been widened in the exploitation for living resources, has broad application prospects.
Accompanying drawing explanation
The infrared figure of erucic acid in Fig. 1: Bruker infrared picture library.
Fig. 2: the infrared figure of embodiment 1 sample.
Fig. 3: the infrared figure of sample in embodiment 2.
Fig. 4: the infrared figure of sample in embodiment 3.
Embodiment
Below in conjunction with drawings and Examples, the present invention is further described.
Embodiment 1
The dry krill 100g of degreasing, adds methyl alcohol 600mL, heating and refluxing extraction 5 times in 70 DEG C of water-baths, and each extraction is filtered for 60 minutes afterwards.Merging filtrate, rotary evaporation is gone out methyl alcohol and is obtained paste 60g.
Get paste 2.00g loading, carry out column chromatography for separation.The sherwood oil of opposed polarity gradient: ethyl acetate (volume ratio is 1:0,50:1,30:1,20:1,15:1,10:1,3:1,1:1,0:1), by polarity gradient elution from low to high.The elution volume of each gradient is 500mL.Collect eluent stage by stage.
Eluent is carried out initial characterization by thin-layer chromatography.Thin-layer developing plate is GF
254high Performance Thin plate (10 × 10cm2), developping agent is dimethylbenzene: ethyl acetate: glacial acetic acid=9:2:0.5, and exhibition distance is 8cm.Qualitative coloring agent used to be massfraction be 5% sulfuric acid solution, dry 5min for 110 DEG C.Merge after launching and only have single point and R
fthe eluent of=0.50, revolves steaming, volatilizes to obtain erucic acid sample.The quality that precision weighing obtains erucic acid sample is 70.8mg.
Get erucic acid sample 5mg, add 20mgKBr, evenly, 12MPa, pressurization 30s obtains transparent compressing tablet in grinding.Detected by MIR-TR, MIR-TR testing conditions: resolution 4cm
-1, sweep time 16s, mid-infrared light source, KBr beam splitter, determines that this sample is erucic acid, and as shown in Figure 2, Fig. 1 is the infrared figure of erucic acid in the infrared picture library of Bruker.Obtain the dry krill of 100g degreasing as calculated and can prepare erucic acid 2.12g.
Embodiment 2
The dry krill 100g of degreasing, adds methyl alcohol 800mL, heating and refluxing extraction 3 times in 75 DEG C of water-baths, and each extraction is filtered for 60 minutes afterwards.Merging filtrate, rotary evaporation is gone out methyl alcohol and is obtained paste 61g.
Get paste 1.66g loading, carry out column chromatography for separation, the sherwood oil of opposed polarity gradient: ethyl acetate (1:0,50:1,30:1,20:1,15:1,10:1,3:1,1:1,0:1), by polarity gradient elution from low to high.The elution volume of each gradient is 500mL.Collect eluent stage by stage.
Eluent is carried out initial characterization by thin-layer chromatography.Thin-layer developing plate is GF254 High Performance Thin plate (10 × 10cm2), and developping agent is dimethylbenzene: ethyl acetate: glacial acetic acid=9:2:0.5, and exhibition distance is 8cm.Qualitative coloring agent used to be massfraction be 10% sulfuric acid solution, dry 5min for 110 DEG C.Merge after launching and only have single point and R
fthe eluent of=0.50, revolves steaming, volatilizes to obtain erucic acid sample.The quality that precision weighing obtains erucic acid sample is 58.8mg.
Get erucic acid sample 5mg, add 20mgKBr, evenly, 12MPa, pressurization 30s obtains transparent compressing tablet in grinding.Detected by MIR-TR, MIR-TR testing conditions: resolution 4cm
-1, sweep time 16s, mid-infrared light source, KBr beam splitter, determines that this sample is erucic acid, as shown in Figure 3.Obtain the dry krill of 100g degreasing as calculated and can prepare erucic acid 2.16g.
Embodiment 3
The dry krill 100g of degreasing, adds methyl alcohol 1000mL, heating and refluxing extraction 2 times in 80 DEG C of water-baths, and each extraction is filtered for 60 minutes afterwards.Merging filtrate, rotary evaporation is gone out methyl alcohol and is obtained paste 62g.
Get paste 1.79g loading, carry out column chromatography for separation, the sherwood oil of opposed polarity gradient: ethyl acetate (1:0,50:1,30:1,20:1,15:1,10:1,3:1,1:1,0:1), by polarity gradient elution from low to high.The elution volume of each gradient is 500mL.Collect eluent stage by stage.
Eluent is carried out initial characterization by thin-layer chromatography.Thin-layer developing plate is GF254 High Performance Thin plate (10 × 10cm2), and developping agent is dimethylbenzene: ethyl acetate: glacial acetic acid=9:2:0.5, and exhibition distance is 8cm.Qualitative coloring agent used to be massfraction be 15% sulfuric acid solution, dry 5min for 110 DEG C.Merge after launching and only have single point and R
fthe eluent of=0.50, revolves steaming, volatilizes to obtain erucic acid sample.The quality that precision weighing obtains erucic acid sample is 63.4mg.
Get erucic acid sample 5mg, add 20mgKBr, evenly, 12MPa, pressurization 30s obtains transparent compressing tablet in grinding.Detected by MIR-TR, MIR-TR testing conditions: resolution 4cm
-1, sweep time 16s, mid-infrared light source, KBr beam splitter determines that this sample is erucic acid, as shown in Figure 4.Obtain the dry krill of 100g degreasing as calculated and can prepare erucic acid 2.19g.
The dry krill of degreasing described in embodiment 1 ~ 3, its water cut is 7 ~ 8%.
Claims (10)
1. an extracting method for degreasing krill erucic acid, is characterized in that, comprises the following steps:
(1) adopted by degreasing krill methyl alcohol heating to extract, removing methyl alcohol, obtains paste;
(2) chromatographic column is got, with sherwood oil wetting dress post after silica dehydrator;
(3) the paste loading in step (1), gradient elution, eluant, eluent is sherwood oil and the ethyl acetate of different proportion, according to polarity wash-out from low to high, and Fractional Collections eluent;
(4) collected eluent is adopted thin-layered chromatography initial characterization, merge after launching and only have single point and R
fthe eluent of=0.50, evaporation, volatilizes to obtain erucic acid sample.
2. extracting method as claimed in claim 1, is characterized in that: in step (1), and described degreasing krill is for adopting hydrophobic solvent by krill except degrease, and described hydrophobic solvent is normal hexane, vegetable oil extractant or sherwood oil.
3. extracting method as claimed in claim 1, it is characterized in that: in step (1), the mass volume ratio of described krill and methyl alcohol is 1g:(6 ~ 10) mL, adopt methyl alcohol heating and refluxing extraction 2 ~ 5 times, Extracting temperature is 70 ~ 80 DEG C, and extraction time is 60min.
4. extracting method as claimed in claim 1, it is characterized in that: in step (2), used silica gel specification is 200-300 order.
5. extracting method as claimed in claim 1, it is characterized in that: in step (3), the volume ratio of described sherwood oil and ethyl acetate is 1:0, (50 ~ 0): 1, and the elution volume of each gradient is 500mL.
6. extracting method as claimed in claim 5, it is characterized in that: in step (3), the eluent petroleum ether of opposed polarity gradient and the volume ratio of ethyl acetate are respectively: 0:1,50:1,30:1,20:1,15:1,10:1,3:1,1:1,1:0.
7. extracting method as claimed in claim 1, is characterized in that: in step (4), and the thin-layer developing plate that thin-layered chromatography uses is GF
254high Performance Thin plate, area is 10 × 10cm
2, developping agent is dimethylbenzene, ethyl acetate and glacial acetic acid mixed solution, and exhibition distance is 8cm.
8. extracting method as claimed in claim 1, is characterized in that: in step (4), qualitative coloring agent used to be massfraction be 5 ~ 15% sulfuric acid solution, dry 5min for 110 DEG C.
9. from a detection method for krill erucic acid, it is characterized in that, comprise the following steps:
S1: adopt the arbitrary described method of claim 1 ~ 8 to prepare erucic acid sample;
S2: the erucic acid sample of gained in step S1 is adopted KBr pressed disc method compressing tablet, gained compressing tablet carries out middle infrared light transmission rate and detects;
Testing conditions: resolution 4cm
-1, sweep time 16s, mid-infrared light source, KBr beam splitter.
10. detection method as claimed in claim 9, is characterized in that: in step S2, tabletting conditions: the mass ratio of sample and KBr is 1:4, pressure 12MPa, duration 30s.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN85103044A (en) * | 1985-10-29 | 1987-05-06 | 贵州省农业科学院中心实验室 | Vegetable seed (oil) erucic acid Fast Detection Technique |
| CN87100646A (en) * | 1987-01-24 | 1987-11-11 | 朱德友 | The erucic acid production technique |
| CA2619858A1 (en) * | 2008-02-22 | 2009-08-22 | Kening Yao | Brassica juncea lines with high oleic acid profile in seed oil |
| CN104049057A (en) * | 2014-05-30 | 2014-09-17 | 中国农业科学院油料作物研究所 | Method for determining compositions of fatty acids and acylglycerols at alpha and beta positions of structured lipid by combining solid-phase extraction and gas chromatography |
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2015
- 2015-07-23 CN CN201510438715.XA patent/CN105067749B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN85103044A (en) * | 1985-10-29 | 1987-05-06 | 贵州省农业科学院中心实验室 | Vegetable seed (oil) erucic acid Fast Detection Technique |
| CN87100646A (en) * | 1987-01-24 | 1987-11-11 | 朱德友 | The erucic acid production technique |
| CA2619858A1 (en) * | 2008-02-22 | 2009-08-22 | Kening Yao | Brassica juncea lines with high oleic acid profile in seed oil |
| CN104049057A (en) * | 2014-05-30 | 2014-09-17 | 中国农业科学院油料作物研究所 | Method for determining compositions of fatty acids and acylglycerols at alpha and beta positions of structured lipid by combining solid-phase extraction and gas chromatography |
Non-Patent Citations (2)
| Title |
|---|
| SYED TUFAIL HUSSAIN SHERAZI等: "Erucic acid evaluation in rapeseed and canola oil by Fourier", 《EUR. J. LIPID SCI. TECHNOL.》 * |
| 马趣环等: "糙叶败酱化学成分研究", 《中药材》 * |
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