KR100663063B1 - Edible flaxseed oil which saturated fatty acid and toxic components were removed therefrom and preparative process thereof - Google Patents

Abstract

Alpha-linolenic acid, which is high in linseed oil, is expected to be used as a serum lipid improving agent, while saturated fatty acids, which are contained in a significant amount, weaken this effect. In addition, trace amounts of cyanogen-glycosides :: linamarine and cyclic-nonapeptide, which are toxic components, are contained in linseed oil. It is desirable to be.

In the present invention, the excess acid in the process of precipitating fatty acids by destroying cyclic nonapeptide, which is a toxic component in the process of alkali saponifying linseed oil to obtain a free fatty acid mixture, and neutralizing the alkali salt of the obtained fatty acid with acid. And cyanogen glycosides are destroyed by heating for a certain period of time, and then extracted with an organic solvent to obtain a free fatty acid mixture. The fatty acid mixture is treated with a urea-complex method to remove saturated fatty acids. The resulting fatty acid is reacted with oxalyl chloride to produce an acid chloride of unsaturated fatty acid, and then reacted with anhydrous glycerin in the presence of a suitable acid-receptor such as pyridine to synthesize triglycerides. And toxic components (cyanogen glyc The present invention relates to linseed oil with the removal or destruction of osides, and cyclic nonapeptides, and with a higher content of (alpha-linolenic acid: (n-3) compared to untreated linseed oil and a method of preparing the same.

Linseed oil, α-linolenic acid, unsaturated fatty acid, cyanide glycoside, fatty acid-urea complex, cyclic-nonapeptides

Description

Edible flaxseed oil which saturated fatty acid and toxic components were removed therefrom and preparative process

1 is a gas chromatogram for the fatty acid composition before the removal of saturated fatty acid of linseed oil,

2 is a gas chromatogram of fatty acid composition after removal of saturated fatty acid from linseed oil.

Figure 3 is a NMR spectrum of linseed oil synthesized after the removal of saturated fatty acids and toxic components.

The present invention provides an edible linseed oil from which saturated fatty acids and toxic components are removed, and a method of preparing the same.

Linseed or flaxseed oil, which has been mainly used in the paint industry, contains a higher amount of alpha-linoleic acid (alpha: n-3) than any other vegetable oils [see Table 1], resulting in hyperlipidemia and hypercholesterolemia. Although flaxseed or linseed oil contains toxic compounds such as cyanogen glycosides such as linamarine and small amounts of cyclic nonapeptides It is known that the use of untreated flax seed or linseed oil for food is prohibited by law. In addition, the linseed oil contains not only alpha-linolenic acid but also a considerable amount (about 10%) of saturated fatty acids (palmitic acid and stearic acid), thereby reducing the serum lipid improvement effect of alpha-linolenic acid. I'm making it.

An object of the present invention is to enhance the effect of improving serum lipids by destroying and eliminating cyanogen glycosides and cyclic nonapeptides harmful to humans in the process of removing saturated fatty acids contained in linseed oil. It is an object of the invention to produce linseed oil which has been toxic and at the same time eliminated toxicity.

Flaxseed or linseed oil contains a significant amount of cyanogen glycosides, such as linamarine, and when fermented linseed, an enzyme called nitrile lyase contained in linseed It is known that it acts on cyanogen glycosides to produce toxic clearing (HCN), so a parent food company in Canada has patented a method of heat treating flaxseed to inactivate nitrile lyase. Supply. However, this patent only inactivates nitrile lyase and does not remove or destroy cynogen glycosides such as linamarine (as found by our experimental results), resulting in HCN poisoning. Not only does not say that the risk of the disease has been completely eliminated, but no countermeasures have been taken against the immunosuppressive activity of another toxic cyclic nonapeptides and serum lipids of alpha-linolenic acid. No measures have been taken to eliminate the saturated fatty acids that weaken the improvement effect.

The lignan component, which is bound to the fiber part of linseed, is known to have phytoestrogen activity, so it is recommended to chew flaxseed whole without eating linseed oil in order to take this component together. Flax seeds, which are licensed as foods, are ingested by many people without the removal of toxic cyanogen glycosides, which may be considered a potential risk of cyan poisoning.

Saturated and unsaturated fatty acids (alpha-linolenic acid, linoleic acid and oleic acid) contained in linseed are often bound to triglycerides of the same molecule, so they are alkali gumed to one glycerin molecule. The saturated and unsaturated fatty acids bound together must be separate molecules. When the fatty acid mixture obtained by hydrolysis is urea-complex, saturated fatty acid is selectively converted into fatty acid-urea complex to precipitate as crystals, thereby separating saturated fatty acid and unsaturated fatty acid by centrifugation or filtration. Cyclic nonapeptides, one of the toxic constituents in linseed oil, are completely destroyed during alkaline saponification of linseed oil, and after acidification, an excess acid is added to precipitate fatty acids by adding acid. Heating can completely destroy and remove cyanogen glycosides, another toxic component. Oxalyl chloride is applied to the unsaturated fatty acid obtained by completely removing saturated fatty acid by urea-complex method to produce an acid chloride of unsaturated fatty acid, and then anhydrous glycerin in the presence of a suitable acid-receptor such as pyridine. It is a main technical problem of the present invention to prepare triglyceride (triglyceride) consisting of only unsaturated fatty acid which is the final substance to be obtained by reaction with.

 Very similar in chemical structure to this peptide as an indirect means of demonstrating complete destruction of the toxic cyclic nonapeptide under the same experimental conditions (alkali concentration, heating temperature and heating time) as saponification of linseed oil. An alkaline substance called bacitracin (cyclic decapeptide) was subjected to alkali saponification under the same reaction conditions, and then contained in trace amounts in linseed as an indicator of the disappearance of antimicrobial activity against Micrococcus luteus (IFO 12708). Indirectly demonstrated the destruction of cyclic nonapeptides, a toxic component known to be present.

To confirm the complete destruction and removal of cyanogen glycosides, hexane extract (flax oil), linseed (hexane extract residue), and fatty acid fractions of linseed and linseed were analyzed. Water extract of acetone powder extracted from raw amateur foil) was used to liberate the clearing (HCN), and then the amount of free clearing was determined by the pyridine-barbituric acid method (Talanta 44, 1203-9, 1997). Plant Science 126,155-6, 1997) and colorimetric was determined at 580nm. As a result, it was confirmed that cyanogen glycosides were not removed from the so-called heat-treated non-toxic flaxseed marketed as a health functional food, and cyanogen glycosides were not contained in the oil extracted from heat-treated flaxseed and raw flax seed with hexane and its hydrolysates. After extracting linseed oil, the remaining residues, that is, flaxseeds, contained cyanogen glycosides at the same level.

Compressing or solvent extracting flaxseed to prepare flaxseed oil; A second step of dissolving linseed oil in a lower alkanol having 1 to 4 carbon atoms and adding a sufficient amount of caustic alkaline aqueous solution to heat and saponify for several hours while at the same time completely destroying a small amount of cyclic nonapeptides; A third step in which after saponification, a sufficient amount of strong acid (hydrochloric acid, sulfuric acid, phosphoric acid, etc.) is added to precipitate a fatty acid, followed by heating for a long time to destroy and remove cyanogen glycosides; The precipitated fatty acid was extracted with an organic solvent, concentrated, and then shaken and cooled by urea-solution dissolved in lower alkanols having 1 to 4 carbon atoms, and cooled to precipitate saturated fatty acid-urea complex as crystals, followed by centrifugation or filtration. a fourth step of removing the -urea complex; Adding a acid to the unsaturated fatty acid fraction to precipitate, extracting with an organic solvent and concentrating to obtain an unsaturated fatty acid mixture; A sixth step of forming an acid chloride of the unsaturated fatty acid by acting oxalyl chloride on the unsaturated fatty acid; And a seventh step of synthesizing triglycerides of unsaturated fatty acids by mixing acid chlorides of unsaturated fatty acids with equivalent anhydrous glycerin and then adding a suitable acid-receptor such as pyridine.

next As an example  The present invention will be described in detail. But this Example  The present invention Limited  It is not.

Example  One

Dissolve 50 ml of ethanol or methanol in 20 gm of flaxseed oil, add about 3.5 gm of caustic soda dissolved in 50 ml of distilled water, install reflux cooling system, and heat for 5 hours to saponify linseed oil. Excess hydrochloric acid or sulfuric acid (e.g. 20 ml of 6N-HCl or 6N-H2SO4) is added, heated for 2 hours, neutralized and concentrated under reduced pressure to recover the alcohol (at this stage Cyanogen glycosides such as linamarine are completely destroyed and removed.) The extracted fatty acid mixture is extracted twice with about 100 ml of hexane, and the hexane layer is distilled to obtain approximately 18.5 gm of various fatty acid mixtures. Separately, 8gm of urea was dissolved in 50 ml of heated alcohol, mixed with vigorous shaking, and allowed to stand at room temperature overnight to precipitate -urea complex of saturated fatty acid as crystals. Centrifuge at 6000 rpm for 15 minutes to remove saturated fatty acid-urea complex crystals. 5 ml of N-HCl is added to the centrifugal supernatant to precipitate unsaturated fatty acid, followed by extraction with ethyl ether, dehydration with anhydrous forget-me-not and distillation of ether to obtain approximately 16 gm of unsaturated fatty acid mixture. This unsaturated fatty acid mixture is mixed with 7.5 gm of oxalyl chloride and left at room temperature for 24 hours to obtain an acid chloride of unsaturated fatty acid. The excess oxarylyl chloride was removed under reduced pressure in a boiling water-water bath, 1.75 gm of anhydrous glycerin and 4.5 gm of pyridine were added thereto and left at room temperature for 3 hours to complete the esterification reaction. The reaction was dissolved in about 100 ml of ethyl ether. After sequentially washing with N-HCl, water, N-NaHCO3 and water, dehydrating with anhydrous forget-me-not, and then distilling ethyl ether, triglycerides composed of only unsaturated fatty acids finally removed by removing saturated fatty acids and toxic components Same as oil) 16.5gm. 3 is an NMR spectrum of the obtained triglyceride. The result of reading the proton peaks of this spectrum is expressed as follows. NMR 300 MHz (TMS, CDCl 3, δ); 0.96 (methyl, 9H, t, J = 7.5 Hz), 1.296 (alipathic methylenes, 36-38 H, br s,), 1.497 (carboxyl adjacent methylene, 6H, t, J = 7.5 Hz), 2.046 (olefine adjacent methylene, 12H, m), 2.302 (= CHC Ha HbCH =, 6H, m), 2.795 (= CHCHa Hb CH =, 6H, m), 4.136 (glyceryl C Ha HbO-, 2H, m, J = 6.0 Hz, 16 Hz ), 4.287 (glyceryl CHa Hb O-, 2H, m, J = 6.0 Hz, 16 Hz), 5.259 (glyceryl CH2 (O-) C H (O-) CH2 (O-), 1H, m, J = 6.0 Hz, 16 Hz), 5.35 (olefinic, = CH-, 14-16H, m)

Example  2

Fatty acid analysis of linseed oil; 20 μl of linseed oil sample was dissolved in 100 μl of 28% sodium-methoxide, 100 μl of hexane was added and mixed well, followed by centrifugation. 5 μl of supernatant was taken, diluted 95 μl of hexane, and 10 μl of the sample was injected into a gas chromatograph. Analyzer: Hewlett packard 5890 II series (HP Co., Wilmington, DE, USA), detector; FID, Column: DB-23 (60m * 0.25mmID, 0.25μm), Oven Temperature: initial 130 ℃, final 230 ℃,, increasement: 2.7 ℃ / min, inlet temp: 270 ℃, detector temp: 300 ℃, carrier gas nitrogen, flow rate: 30ml / min

Table 1. Fatty acid composition ratio (%) of vegetable oil Analysis

 Table 1 shows the analysis of fatty acid content in various cooking oils, including linseed oil, available in the domestic market. 1 and 2 are gas chromatograms before and after removing saturated fatty acid from linseed oil. 1 shows ① palmitic acid, ② stearic acid, ③ oleic acid, ④ linoleic acid, and ⑤ α-linolenic acid. Palmitic acid and stearic acid peaks observed in FIG. 1 are not observed in FIG. 2. Fig. 2 is a gas chromatograph after removal of saturated fatty acid in linseed oil, where ①oleic acid, ②linoleic acid, and ③α-linolenic acid. Table 2 shows the content ratios of the various fatty acids calculated by the peak area integration values of FIGS. 1 and 2.

 Table 2 is a table comparing the fatty acid composition of linseed oil and linseed oil removed from saturated fatty acids.

Example  3

Linseed  Acid value, peroxide value, and Okdo  Measure;

The acid value, peroxide value, and iodine value of the flaxseed oil from which the saturated fatty acid and the toxic component obtained by the method of Example 1 were removed to determine whether the flaxseed oil obtained by the method of Example 1 was obtained with edible oil. Was measured by the method registered in the Republic of Korea Food Code. The results are shown in Table 3 and the results are not in an inedible range.

Example   4

Linamarin  Etc Cyanoglycosides  Content analysis;

In order to analyze the cyan content in raw flax and edible heat-treated linseed prepared by a commercially known method, and the flaxseed oil obtained by removing the saturated fatty acid and toxic components obtained in Example 1 of the present invention, these samples were subjected to nitrilease. (nitrile lyase) and linamarinase mixed enzyme (Plant Science 126, 155-62, 1997) to free the cyanide free pyridine barbituric acid method (Talanta 44, 1203-1209. 1997, Plant Science 126, 155-162. 1997), absorbance at 580 nm was measured, and was separately released by applying the same mixed enzymes (nitrile lyase and linamarinase) to a certain amount of standard linamarine (AG Scientific, Inc., USA). Cyanogen-glycosides content in the sample was determined by preparing a straight calibration curve in the range of 10-50 μg / 6.2 ml (final) by bleaching HCN in the same way. The. Nitrile lyase and linamarinase enzyme solutions were homogenized with a small amount of raw amylase in hexane and degreased to form acetone powder for the residue, and then a predetermined amount of 0.1M-acetate-buffer (pH 5.6). The same buffer solution was repeatedly dialyzed at low temperature with a dialysis external solution to completely remove cyan comp, and used as an enzyme solution. Clearing analysis values are shown in Table 4 in terms of linamarine content. As shown in Table 4, most of the cyanogen glycosides, which are so-called patented food products that are licensed for food, remain unremoved or destroyed, and cyanogen compounds in the linseed oil from which hexane is extracted from raw or heat-treated linseed. Flaxseeds extracted from linseed oil do not contain cyanogen glycosides and contain most of the cyanogen glycosids contained in linseed, regardless of whether they are heat-treated or raw.

Example  5

Cyclic Nonapeptide (Cyclic nonapeptide Inactivation experiments;

Experiments demonstrating the inactivation of cyclic nonapeptides, an immunotoxic component contained in linseed oil obtained in Example 1, adopted an indirect method. That is, although cyclic nonapeptides contained in linseed oil have been reported to have immunotoxicity, they are not easy to isolate because they are trace components at levels of 10-4% and are not commercially available as reagents. The experiment was carried out using a substitution of bacitracin, a peptide antibiotic with a very similar chemical structure to cyclic nonapeptides. In order to know whether this substance is completely destroyed under the same conditions as alkaline saponification of linseed oil of Example 1, the antimicrobial activity of baccitracin against Micrococcus luteus was determined using a liquid medium dilution method. The IC 50 value was indirectly verified to destroy and inactivate cyclic nonapeptide, an immunotoxic component. The measured IC50 values are shown in Table 5.

Table 5 shows the antimicrobial activity of the antibiotic bacitracin as an indirect method to demonstrate the complete destruction of toxic nonapeptides during saponification of linseed oil.

Example  6

A glycerol content test was conducted to confirm that the final product obtained in Example 1, that is, "saturated fatty acid and toxic component removed linseed oil" was prepared in triglyceride form. Glycerol content was analyzed using a commercially available triglyceride assay kit (enzyme method, Youngdong auto-kitage kit). This kit is used to analyze triglyceride in serum lipids which are colorimetrically analyzed by sequentially developing and developing lipase, glycerol kinase, glycerol-phosphate-oxidase, and peroxidase in serum lipids. Instead of using the included lipase, it was chemically examined, neutralized, and then analyzed using the enzymes contained in the kit. Separately, a calibration curve for this method was prepared using 99.7% -glycerol, and the glycerol content was analyzed for linseed oil and “flaxseed oil without saturated fatty acids and toxic components” prepared in Example 1, and the results are shown in Table 6. Indicated.

Table 6 shows the analysis of glycerol content of linseed oil, linseed oil removed from saturated fatty acids and toxic components.

 The present invention removes saturated fatty acids contained in linseed oil to increase the content of alpha-linolenic acid (n = 3), and cyclic-nonapeptides and cynogen glycosides which are harmful to humans. A novel linseed oil is provided which enhances the effect of improving serum lipids by destroying and eliminating cynogen glycosides and at the same time eliminating toxicity.

Claims (2)

 Flaxseed oil composed of triglycerides of unsaturated fatty acids by removing saturated fatty acids and cyclic nonapeptides and cyanogen glycosides. 1 compression or solvent extraction of linseed to prepare a flaxseed oil (flaxseed oil); 2. dissolving the obtained linseed oil in lower alkanols having 1 to 4 carbon atoms, saponifying by adding caustic alkali, and breaking the trace-containing cyclic nonapeptide; 3. Add the strong acid selected from hydrochloric acid, sulfuric acid and phosphoric acid to neutralize the caustic soda used for saponification to the saponified product obtained in the second step to precipitate fatty acid and heat for a certain time to obtain cyanogen glycosides ( destroying and removing cyanogen glycosides); 4. The precipitated fatty acid obtained in the third step was extracted with an organic solvent, concentrated, and added thereto with urea dissolved in a lower alkanol having 1 to 4 carbon atoms, followed by saturated fatty acid-urea complex. Precipitating and separating into crystals to remove the saturated fatty acid-urea complex; 5. adding an acid to an unsaturated fatty acids-urea complex dissolved in the solution to precipitate, followed by extraction and concentration with an organic solvent to obtain an unsaturated fatty acid mixture; 6. acting oxalyl chloride on the unsaturated fatty acid mixture to obtain an acid-chloride of unsaturated fatty acid; And 7. Glycerol and acid chlorides of unsaturated fatty acids are reacted in the presence of a suitable acid-receptor, followed by post-treatment to prepare triglycerides containing unsaturated fatty acids, which are completely free of saturated fatty acids and completely destroyed or eliminated of toxic components. How to.

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