WO2004048419A9

WO2004048419A9 - Method of classifying, purifying and using fenugreek seeds

Info

Publication number: WO2004048419A9
Application number: PCT/JP2002/013432
Authority: WO
Inventors: Toshihisa Tanaka; Kenji Iwamoto
Original assignee: Air Green Co Ltd; Toshihisa Tanaka; Kenji Iwamoto
Priority date: 2002-11-28
Filing date: 2002-12-20
Publication date: 2004-12-29
Also published as: JPWO2004048419A1; AU2002368394A8; JP2005341801A; WO2004048419A1; AU2002368394A1

Abstract

Fenugreek seeds have been employed for edible use as well as a material for extracting galactomannan, essential oil, oleoresin, saponin, proteins, 4-hydroxyisoleucine, etc. therefrom. These substances are extracted exclusively from seeds. In case of chemical extraction, however, all of these substances can be hardly extracted at the same time, since different extraction solvents should be employed for galactomannan and other substances. In addition, a large amount of an alcohol is needed in extracting galactomannan, which makes the product expensive compared with other galactomannan-type thickeners such as guar gum. A method of preparing an albumen powder containing galactomannan at a high purity which comprises separating albumen from seed coat with a grinder mill by taking advantage of the difference in hardness between albumen and seed coat, then milling the albumen in an impact mill or an air-stream mill to give particles with different sizes and then classifying them through a sieve. Thus, it becomes possible to separate galactomannan exclusively by the physical procedures. As a result, all effective components contained in seed coat can be obtained simultaneously and the production cost of fenugreek galactomannan can be successfully cut down.

Description

Description: Separation, purification and utilization of fenugurik seeds

The present invention provides a method for physically separating fenugreek seeds into endosperm and seed coat (including cotyledons), physically refining the endosperm, and producing an endosperm powder containing high-purity galactomannan. It is a method of using huengreek seed coat (including cotyledons) and endosperm as various industrial raw materials including extraction. Background art

Fenugreek (Trigonella Foenum Graecum) is a one-year grass of the legume family, and its seeds have been used for food and medicine since ancient times. The most famous use is as a raw material for curry, used as a spice to produce the aroma and stickiness of curry. The main component of fenugreek endosperm is galactomannan (see Fig. 1), which is a high molecular polysaccharide that binds galactose (see Fig. 2) and mannose (see Fig. 2) at a ratio of about 1: 1. Because it is a polysaccharide that humans cannot digest, it is generally called dietary fiber. Galactomannan has the property of absorbing water and gelling when dissolved in water, and is widely used as a natural thickener as an industrial raw material for foods, cosmetics, pharmaceuticals, and the like. Other galactomannan-based natural thickeners include guar gum and mouth-to-cast bean gum, but compared to these gums (a substance that has a thickening effect is referred to as gum in English), fenugeek gum (endosperm) ) Has the highest proportion of galactose. Galacto in galactomannan It has the property of improving water solubility, emulsifiability and stability as the ratio of water increases. The ratio of galactose to mannose is 1: 2 for guar gum, 1: 4 for locust bean gum, and 1: 1 for fenegriek gum. Whereas other gums need to be warmed or agitated longer when dissolved, fenegreak gum has the excellent properties of quickly dissolving in water at room temperature to form a stable gel. . Therefore, it is receiving much attention as a raw material from various fields. Fenugreek seeds have long been used as medicinal plants that have been used only as edible spices for nutritional tonicity, increased appetite, antipyretics, increased lactation, and reduced blood sugar. These effects are caused by galactomannan contained in endosperm and various components contained in the seed coat. A commonly used effect in the Middle East and India is the promotion of lactation by Fenugreek. The Fuenuguriku, steroidal saponin (see FIG. 3) are included many, _c Jiosugenin there is Jiosugenin typical Furoosutanoru type saponins therein is industrially, female hormones progesterone synthesis Taking fenegreek seed, a precursor, is thought to be converted into progesterone in the body, which in turn promotes lactation. For this reason, women in the Middle East and India are encouraged to eat fenugreek seeds after childbirth. In addition to such medicinal effects, saponin has industrial value as a foaming agent and an emulsifier. In addition, fenugurik seeds have a lipoprotein-lowering effect and a hypoglycemic effect in cholesterol. Many of these research reports have been published. Only the documents at hand include the following.

Cholesterol lowering action: RD Sharma, Hypercholesterolemic activity of fenugreek (T. foenum graecum) An experimental study in rats, Nutrition Report International, July 1984 Vol.30 No.1.

A.J.Evans, R 丄 .Hood, D.G.Oakenfull, G.S.Sidhu, Relationship between structure and function of dietary fiber: a comparative study of effect of three galactmannans on cholesterol metabolism in the rat.

RD Sharma, An evaluation of hypercholesterolemic factor of fenugreek PRPU Rao; B. Sesikeran, PS Rao, AN Naidu, VV Rao, EP Ramachandran, Short term nutritional and safety evaluation of fenugreek, Nutrition Research Vol. 16 No. 9, pp 1495- 1505, 1996

A. Neeraja, P. Rajyalakshmi, Hypoglycemic effect of precessed fenugreek seed in human, Food Sci. Technol, 1996, Vol. 33, No. 5, 427-430

Hypoglycemic action:

R.D.Sharma, T.C.Raghuram, Hypoglycemic effect of fenugreek seed in non-insulin dependent diabetic subjects, Nutrition Research Vol. 10, PP.731-739, 1990.

G. Ribes, Y.Sauvaire, C. Da Costa, J.C.Baccou, Antidiabetic effects of subfractions from fenugreek seeds in diabetic dogs, Proceedings of the society for experimental biology and medicine, 182, 159-166, 1986.

Z. Madar, Fenugreek (Trigonella Foenum Graecum) as a means of reducing postprandial clucose level in diabetic rats, Nutrition Reports International, June 1984 Vol. 29 No. 6

A. Bordia, SK Verma, KC Srivastava, Effect of ginger (Zingiber officinale Rose.) And fenugreek (T rigonella foenum graecum L.) on blood lipids, blood sugar and platelet aggregation in patients with coronary artery disease, Prosaglandins, Leukotrienes and Essential Fatty Acids, 1997, 56 (5), 379-384 Dietary fiber usually has a more or less lowering effect on cholesterol and triglyceride, and naturally, fenugreek endosperm also has these effects. An effect that is not found on other dietary fibers in the fenegriek species is its hypoglycemic effect. Initially, a number of studies have suggested that galactomannan contained in the fenugreek endosperm is involved in the hypoglycemic effect. It was found to contain amino acids, which acted to increase insulin secretion and lower blood glucose.

Y. Sauvaire, P. Petit, C. Borca, M. Manteghetti, Y. Baissac et. Al " 4-Hydroxyisoleucine, A Novel Amino Acid Potentiator of Insulin Secretion, Diabetes, Vol. 47, February 1998.

Since the galactomannan of fenugreek is also considered to have a sugar absorption inhibitory effect, it is considered that fenugreek suppresses the rise of blood sugar by multiple actions. Furthermore, the fenugreek seed coat contains a resin part and a large amount of a tank which are used as a fragrance of the power ray, and can be used as an industrial raw material in many fields by extracting these.

Huenegeek seeds have a completely different composition between endosperm and seed coat. Endosperm is mostly made of a polysaccharide called galactomannan, while seed coat is composed of essential oils, oleoresins, saponins, and proteins. Made of, cellulose, hemicellulose, lignin, etc. Galactomannan is a high-molecular-weight polysaccharide in which galactose is attached to the linear polymer of mannose by the side chain, and has the property of increasing the viscosity when dissolved in water. It is widely used as an emulsifier in food, cosmetics and other industrial raw materials. However, the use of fenugreek galactomannan is less than other galactomannans. The small seeds make it difficult to remove the endosperm, and the unique aroma and bitterness alter the taste and flavor of food when mixed with food. Normally, the method of extracting galactomannan from seeds involves powdering the seeds and then chemically treating them. This method utilizes the property that galactomannan dissolves in water but does not dissolve in alcohol. First, powdered seeds are put into water to elute galactomannan. Next, the residue is removed by filtration or centrifugation, and alcohol (mainly ethanol) is added to the remaining aqueous solution of galactomannan until a concentration of about 50% is obtained. Kutmannan causes precipitation. By filtering the solution, a precipitate of galactomannan can be obtained. The resulting galactomannan is colored brown and smells, so it is decolorized and deodorized again using alcohol to obtain white galactomannan.

The disadvantage of this method is that large amounts of alcohol are required. Since galactomannan has a strong viscous action, it becomes messed up in a 30/0 solution and loses its fluidity. Therefore, the residue cannot be removed by filtration or centrifugation. For filtration and centrifugation, the flowable concentration of 1-2Q / is limited. For example, assuming that galactomannan is eluted from fengreat seeds in 1 L of water to obtain a 2% aqueous solution of galactomannan, the same amount of alcohol is required to precipitate galactomannan. And the resulting galactomannan is 20 g. In addition, alcohol is used for washing, so alcohol that is more than 50 times the amount of galactomannan actually obtained is required. If this method is used, the cost of alcohols, especially ethanol, is high, and it is impossible to produce galactomannan on a commercial basis. (Commercial base is the price that can compete with guar gum and mouth-to-mouth bean gum)

In addition to galactomannan, fenugreek seeds contain fenugreek oil (essential oil), fenugreek oleoresin (resin), saponins, proteins, etc. It is done in. In particular, fenugreek oil and fenugreek oleoresin are widely used as fragrances. These extractions are all performed by crushing seeds. Also recently talked about Extraction of 4-no- and hydroxy-isoleucine, insulin-secreting thighs that are considered to be involved in the hypoglycemic effect of fenegurik seeds, is also performed from seeds. (US patent 5,470,879)

According to the patent for the method of manufacturing 4-hydroxyisoleucine, it is very similar to the conventional method of extracting savon and the like. Figure 5 shows a flow chart of our extraction method for saponins. This is done by extracting phenoglycoyl from fenegriek seeds using hexane as a solvent, extracting the residue with 70% ethanol, concentrating it, placing it in 95% ethanol to precipitate saponin, and filtering. Then, saponin is removed, and the remaining solution is concentrated to obtain fenuguri oleoresin. Furthermore, the residue is extracted with an alkaline aqueous solution and neutralized to precipitate the protein and remove the protein. It is unlikely that all this extraction will be done at the same time, but individual extractions will be made from Huenig seeds based on demand. The following documents describe such extraction methods.

A. Benichou, A. Aserin, N. Garti, Steroid-Saponins from fenugreek seeds: Extraction, Purification and Surface properties, J. Dispersion Science and Technology, 20 (1 & 2), 581-605, 1999.

N. Garti, A. Aserin, B. Sternheim, Fenugreek Galactomannans ad Food Emulsifiers, Lebensm.—Wiss, u-Technol., 30, 305-311, 1997

WS Taylor, MS Zaman, Z. Mir, RS.Mir, SN Acharya, Analysis of Steroidal Sapogenins from Amber Fenugreek (Trigonella foenum gracecum) by Capillary Gas Chromatography and Combined Gas Chromatography / ass Spectrometry, 丄 Agric. Food Chem. 1997, 45 , 753-759.

RR.Petit, YDSauvaire, DM Hillaire-Buy et.al., Steroid saponins from fenugreek seeds: Extraction, purification, and pharmacological investigation on feeding behavior and plasma cholesterol, Steroids 60: 674-680, 1995 I. Blank, J. Lin, S. Devaud, R. Fumeaux, The Principal Flavor Compounds of Fenugreek (Trigonella foenum gracecum L), 997 Ameican Chemical Society, Chapter 3.

MK Osman, LS.Simon, Biochemical studies of some non-conventional sources of protein Part 5.Extraction and characterization of protein from fenugreek seed (Trigonella foenum) Die Nahrung 35, 1991, 3, 303-308

4-Hydroxyisoleucine is extracted by passing a 70% alcohol extraction solution through a cation exchange resin and a silica gel adsorption column to isolate 4-/-hydroxyisoleucine. This extraction method itself is a method that has been commonly used in the past, without any novelty. After isolation of 4-hydroxyisoleucine, the above method can be carried out to extract the same tree, savonin and the like. When chemically extracting all active ingredients from fenegurieg seeds, the method of extracting galactomannan is completely different from the method of extracting saponin, 4-hydroxyisoleucine, and the like. Galactomannan is an extraction with water, while saponin and 4-hydroxyisoleucine are with an alcohol solution. If galactomannan is first extracted with water, saponin, 4- /, and hydroxy-isoleucine are also dissolved in the extract. To add ethanol to a 2% galactomannan aqueous solution to make a 50% ethanol solution and precipitate galactomannan), and to make a large amount of the remaining extract into a 70% ethanol solution or a 95% ethanol solution, use a large-scale concentrator. And a lot of ethanol again. With the method of extracting galactomannan with water, it is impossible to extract saponin, 4-hydroxyisoleucine, etc. industrially at low cost. After all, if galactomannan is first extracted from the fungigreek seed, saponin and 4-hydroxyisoleucine cannot be extracted. Conversely, it seems better to extract saponin, 4-hydroxyisoleucine, etc. first, and then extract galactomannan.In this case, the extraction of galactomannan is described in Fig. 5. Must be performed prior to protein extraction. However, Since the water extraction itself requires a large amount of alcohol, it does not solve the problem of inexpensively isolating the active ingredient contained in fenugreek seeds.

Eventually, we conclude that the use of water in the processing of Huenegreek seeds is contraindicated. Disclosure of the invention

We decided to consider an extraction method by physical treatment as a method to obtain galactomannan without using water. The method initially thought was to scrape the surface of the seed, and thought that if the seed coat was scraped, endosperm would come out. If the seed coat is not discharged at the same time as the seed coat is removed, only the endosperm remains. Therefore, as a method for shaping the seed coat, a device as shown in Fig. 6 in which a propeller is installed at the bottom of a cylindrical wire mesh (peeling off) Device). The seeds are put into it, and the propellers are rotated by a motor to rotate the seeds at high speed in a cylindrical wire mesh, and the seeds come into contact with the wire mesh and the seed coat on the surface is shaved. I thought. The diameter of the wire mesh was 60 mesh of 250 microns. This is because the size of the endosperm was assumed to be larger. Shaved seed coats (including cotyledons) are discharged from this wire mesh when they become smaller than 250 microns. As a result of the experiment, the seed coat and endosperm were separated by rotating for about 2 hours. This is due to the difference in hardness between the seed coat, cotyledon and endosperm, with cotyledon being the softest, followed by seed coat and endosperm. After placing the seeds, the seeds are rotated in the cylindrical wire mesh of the peeling device, so that the wire mesh and the seeds rub each other, the seed coat is shaved, and the seed coat becomes thin to some extent. When the seeds collided with each other and collided with the wire mesh, the seeds were broken, the endosperm and cotyledons jumped out, and the seed coat and cotyledons that were exposed were discharged as small particles out of the wire mesh. It is based on the principle that only the endosperm remains in the wire mesh. Because the endosperm is hard and cannot be easily scraped off with this peeling device, only the endosperm remains.

The next thought was that putting the broken seeds rather than putting the seeds in the peeling device might help the seeds separate faster. The seeds have an oval shape and therefore have low resistance, but we thought that broken seeds would be separated quickly because they are uneven and resistant. So we tried to crush the seeds with various commercially available crushers. In some mills, the seed coat and endosperm, which have high crushing ability, were broken finely and came out of the wire mesh, so they could not be sorted by the peeling device. With another mill, only the surface was shaved and slippery, which took a very long time to grind. After several experiments, the crusher (mill type crusher) was able to pulverize moderately, and the seed coat and endosperm were separated. The milled seeds were a mixture of seed coat and endosperm. The seed pieces were placed in a peeling device, and the seed coat and endosperm were separated. As a result, the seed coat and endosperm were separated in about an hour of rotation.

Even though most of the seed coat could be separated from the endosperm, there was endosperm in which part of the seed coat was attached to the endosperm, and without removing it, it was not possible to obtain an endosperm powder with a high galactomannan content. So, I tried to remove a part of the attached seed coat by rotating the peeling device further. By rotating it for about 100 minutes, the seed coat can be removed to some extent, but it has the disadvantage of requiring a long processing time. It is the lens of endosperm This is because when the seed coat has a shape like that and the seed coat adheres to the convex part, it can be removed quickly, but it is difficult to remove it when the seed coat adheres to the concave part.

So, I thought that it would be possible to sort endosperm using a color sorter. Hu X Nugreek The seed coat is brown, and the endosperm is light brown. For this reason, endosperm with attached seed coat appears darker with a color sorter, and endosperm without seed coat power appears white, and can be sorted with a color sorter. The color-separated endosperm with no attached seed coat was placed in a peeling machine, rotated for about 30 minutes, and the surface was thinly exposed to obtain a fine endosperm powder with a galactomannan purity of 80% or more.

Endosperm before being subjected to a color sorter is mixed in various sizes of about 0.5mm to 2mm. The color sorter recognizes each of these endosperms with a camera, and when it recognizes dark endosperm, it shoots off only that endosperm with an air gun. Using a color sorter separates the endosperm with and without the seed coat and increases the efficiency of the subsequent peeling treatment. The problem was that the processing capacity of the color sorter was low. Increasing the processing capacity (moving speed) reduces the ¾IJ ability, and decreasing the processing capacity improves the sorting ability. With the optimal sorting capacity, only a few kilograms can be processed per hour. The monthly production capacity was about 700 kg. When the endosperm to which a part of the seed coat was attached was exposed with a peeling device, a long processing time was required. In light of the above, in order to increase productivity with this method, it was necessary to take some measures such as improving, changing, and adding mechanical equipment.

Another solution has been found in the process of powdering the endosperm. Our Huen Greek The specification of endosperm powder originally stipulated a particle size of 100 mesh (150 im) or less. F: E-Nugreek endosperm was too hard and resilient to be powdered with a mortar or impact mill. Therefore, it was decided to pulverize it with the most powerful air-flow-type pulverizer currently considered. We used a pneumatic pulverizer called Selenium Mirror (trademark) manufactured by Masuko Sangyo Co., Ltd. (Masuyuki Sangyo Co., Ltd .: 1-12-24, Honcho, Kawaguchi-shi, Saitama) An air-flow crusher is comprised of a plurality of high-speed rotating star-shaped metal plates called rotor blades in a cylindrical tube. By blowing a jet stream into the cylinder and rotating the metal plate, a swirling vortex of air is generated in the cylinder at high speed, causing effects such as impact 'shearing', 'compression', 'milling' and high frequency vibration. Is a crusher for pulverizing the powder. Crushers that perform this function are available from several companies. (See Fig. 7) However, even with this crusher, only 30% of Fenugreek endosperm can be reduced to a powder of 100 mesh or less at one time, and it must be passed through three times to obtain a powder of 100 mesh or less. Needed.

When the process of crushing once, passing through a 100-mesh sieve, further crushing, and passing through a 100-mesh sieve is repeated, the color of the powder passing through the 100-mesh sieve and the color of the remaining powder are different. Noticed. This is the result of the force that the seed coat part attached to the endosperm was finely crushed first and the endosperm part was not sufficiently crushed. This was attributed to the difference in hardness between the seed coat and the endosperm.

Fenugreek endosperm is in the form of a lens and has irregularities. When the surface is peeled off by a peeling device, the convex side is easily peeled off, but the concave side is hardly peeled off. Therefore, the peeling device cannot completely remove the seed coat. To solve this problem, embryos When the milk is crushed into particles, endosperm particles and seed coat particles are produced, and the seed coat is softer, resulting in smaller particles, and endosperm becomes larger particles than the seed coat. As a result, it was found that the seed coat and endosperm could be separated by a sieve.

Next, the endosperm that had not been sorted by the color sorter, that is, the endosperm containing the endosperm with the seed coat portion mixed, was passed through an air-flow crusher at a low rotation speed. The resulting powder

When passed through a 100 mesh sieve, 20% passed through the sieve and 80% did not. The powder passed through the sieve had a galactomannan content of 40-60%. The color was brown and odory and the taste was bitter. When the powder that did not pass through the 100 mesh was passed through the air-flow type powder again at a low rotation speed, 10% passed through a 100 mesh sieve and 90% did not pass through the sieve. The galactomannan content of the sieved powder was 60-80%. The powder that did not pass through the sieve had a galactomannan content of 80% or more. As a result, it was confirmed that phenogreek endosperm powder containing galactomannan with a high purity can be produced by crushing fenegriek endosperm and separating the powder according to the difference in particle size. This method was clearly more efficient than the method of peeling the surface. Finally, when crushed at the maximum number of revolutions, 90% of the powder passes through 70 mesh, and this is the final product.

The air-flow type mill has a drawback that the pulverizing capacity is high but the processing capacity is small. In the first and second pulverization, the rotation speed is reduced to produce weak powder, so the pulverizing capacity is lower than that of the air-flow type pulverizer, but the throughput is large, and it may be possible with the impact type pulverizer. I thought and decided to give it a try. The impact-type pulverizer crushes the raw material by giving an impact to the raw material, and does not blow the jet stream like the air-flow-type pulverizer. Our use In the crusher, a large number of knurls are mounted around a disk-shaped metal plate, and the disk rotates at high speed so that the hammer crushes the raw material. Various types of impact crushers are commercially available. (Refer to Fig. 8) The results of the experiment show that in the first grinding, 10% passed through a 100-mesh sieve, and 90% did not. In the second grinding, 25% passed through a 100 mesh sieve and 75% did not pass. The ratio was about 30%, which is the same as that when weakly pulverized with an air-flow type pulverizer. The powder that did not pass had a galactomannan content of 80% or more. As a result, it was found that the impact-type pulverizer can be used to remove the seed coat. Since the impact crusher has five times the processing capacity of the air crusher, it has been found that the combined use of the impact crusher and the air crusher can dramatically improve the throughput. Also, the impact type crusher has an advantage that the price is lower than that of the airflow type crusher.

The remaining problem is the separation of seed coat and endosperm from the seed. In this case as well, we thought whether it could be separated from the difference in hardness between the seed coat and endosperm. However, any crusher is made for the purpose of making powder. The performance depends on how fine the powder can be made. However, the only way to grind the Huen Greek seeds is that the seed coat is broken, and the endosperm comes out without breaking and can be further separated. I thought that a simpler crusher rather than a very high-performance crusher would be more suitable, and after examining various crushers, a mortar-type crusher for sawing was found. I decided to try. The machine tested was a slate mill (trademark) from Nishimura Seisakusho Co., Ltd. (9 each of Matsuyama-cho, Yao-shi, Osaka). The principle is the same as a stone mill. It is. The two mortars with metal grooves face each other with a certain gap, and the inner mortar rotates to grind the raw material, and the crushed raw material is discharged to the outside through the groove It works. (See Fig. 9)

The difference from ordinary millstones is that millstones are milled by turning the upper and lower mills in close contact and produce powder, while milling mills are installed with a certain distance between the upper and lower mills. Is not closely related. The principle is that if a certain interval is left, particles smaller than that interval cannot be formed. When crushed by this crusher, a granular powder such as coarsely ground coffee beans can be produced. 18-30 mesh for this crusher

(0.5-0.85 mm) stitches were connected. The reason for adding | chang to the mesh size of the sieve is that the smaller the mesh size, the finer the particles of the seed coat that are discharged, the greater the number of passes through the crusher, and the slower the processing speed. Conversely, when the mesh size is coarse, the number of times of passing through the grinder is reduced, and the processing speed is increased, but the small endosperm passes through the sieve and the yield is reduced. Since the mesh size of the sieve is related to the characteristics of the seeds of the raw material and the production yield, it is necessary to adjust the mesh size as needed. We determined that the smallest endosperm was 0.5 mm, so the mesh size was between 18-30 mesh.

The experiments were performed on sieves with a mesh size of 0.7 mm, but the throughput of the grinder was high and our machine processed 120 kg of seeds per hour. When the mortar spacing was set to a minimum of 0.48 mm, 40% of the seed coat and cotyledons passed through the sieve in the first pulverization, and the endosperm was not broken at all and did not pass through the sieve in the form of a lens. Was discharged. Endosperm is hard but elastic and shaped like a lens, so it has a certain width However, because of the lack of thickness, it is thought that they would pass sideways and slip through the gap between the mortars. The seed coat is crushed because it is soft and brittle, but the endosperm is thought to pass through in its shape without being crushed due to its elasticity. In the second milling, 40% passed through the sieve again, and the remaining 60% that did not pass through were all endosperm, almost completely separating seed coat and endosperm. As a result, a pulverizing production method with a much higher processing capacity was established compared to the production of refined fengreat endosperm powder using a peeling device that had been used previously.

Endosperm powder made from seeds accounts for 20-25% of the seeds, of which high-purity endosperm powder with a galactomannan content of 80% or more accounts for 70%. The remaining 75-80% is seed coat and cotyledons. The method of separating seed coat and endosperm only by a physical method without performing water extraction in this way can produce galactomannan at low cost without using a large amount of alcohol. Furthermore, galactomannan is not contained in the seed coat obtained as a by-product. That is, without extracting galactomannan, extract many active ingredients, fenegrig oil, fenegrig oleoresin, fenegrig saponin, Φhydroxyisoleucine, and fenugreek tank, which are contained in fenegreeg seeds. It became possible. By extracting various active ingredients from the seed coat, not from the seeds, it became possible to obtain all the active ingredients contained in Fenugreek seed power << efficient and less expensive. The method for extracting the active ingredient from the seed coat is impossible without the present invention for separating the seed coat and endosperm, and is a new invention accompanying the present invention.

In addition, by crushing the seed coat into powder, it becomes a flavoring that adds curry flavor to food etc. Can be used. In addition, the medicinal properties of penugreek are caused by substances contained in the seed coat in large amounts, and by taking it, various health-enhancing effects can be obtained. It can also be used as a raw material. As a raw material for functional foods, health foods and the like, fengreak endosperm powder can also be used. Galactomannan contained in endosperm has the property of swelling and gelling when dissolved in water, and is a food that humans cannot digest. Dietary fiber itself has the effect of an inflatable laxative, which can relieve constipation, and flapping can provide a feeling of fullness. It can be used as a diet food material. In this way, by separating fenugreek seeds into endosperm and seed coat, the utility value of fenugreek seeds is increased and it can be used as an industrial raw material for many purposes.

"Fenugu Greek Endosperm Powder" Standards and Test Methods

[Properties]

Endosperm powder A: Light brown powder with slightly animal odor.

Endosperm powder B: Light brown powder with a slight smell.

[Confirmation test] A, B common

(1) Add 2 mL of this product to 4 mL of isopropyl alcohol, moisten it, add 200 mL of water with vigorous stirring, and stir until dispersed evenly to form a viscous liquid. Stir this solution in a water bath for about 10 minutes, then cool to room temperature Sometimes the viscosity is almost the same as before heating.

(2) Add 2 mL of sodium (1 → 20) to 10 mL of the viscous liquid obtained in (1), mix and leave it in a jelly form.

Only A

(3) When 0.5g of this product is dissolved in an appropriate amount of water, dissolve within 5 minutes. The dissolved jelly-like substance has no odor or bitterness.

(1) Acid insolubles A: 3% or less B: 5% or less

Precisely weigh about 2.0 g of this product, place it in a beaker containing 150 mL of water and 15 mL of sulfuric acid, cover with a watch glass, and heat on a water bath for 6 hours. Occasionally wash away the sample on the beaker wall with a glass rod to compensate for water loss due to evaporation. After cooling, accurately weigh about 500 mg of diatomaceous earth for filtration, which has been dried at 105 ° C for 3 hours, add to this solution, mix well, and filter using a glass filter of known weight. Wash the residue on the filter 3 times with hot water, dry it at 105 ° C for 3 hours and weigh. Subtract the weight of the glass filter and the diatomaceous earth for filtration from the weight obtained at this time.

(2) Water-insoluble substances A: 3% or less B: 5% or less

Precisely weigh about 1 g of this product, add 199 mL of water, and stir vigorously for 10 minutes. Then, mix gently for 3 minutes. This solution is placed in an Erlenmeyer flask, stoppered, and heated on a water bath at 30 ° C for 3 hours. Remove 50 g of solution and centrifuge at 3000 rpm for 30 minutes You. Remove the supernatant, add 50 mL of purified water, stir, and centrifuge again under the same conditions. Repeat this operation twice. Remove all supernatant, dry at 105 ° C for 2 hours, and weigh the residue.

(3) Common for heavy metals A and B 20 ppm or less

Remove 1 g of this product, conduct the test by operating the Japanese Pharmacopoeia 13, general test 23, and heavy metal test 2 according to the second method. To the comparative solution, add 2.0 mL of the standard solution.

(4) Arsenic A, B common 2 ppm or less

Prepare 1 g of this product, prepare a sample solution according to the Japanese Pharmacopoeia 13, General Trial! ^ Last 41, Arsenic 1¾ method 3, and perform the test using the device B. [Loss on drying] A, B common 10.0% Below (1g, 105. C, 5 hours)

[Viscosity] (Intrinsic viscosity of 1 w / v% solution)

A: 3000 mPa's or more

B: 1000 mPa's or more

Perform a retest on the 1% solution of this product according to JP13, General Test Method 36, and Viscosity Test Method. (Grain 闳 100 mesh or less (B) 7Ό mesh or less (A)

[Nutrition test]

(1) Protein 5.0% or less

Precisely weigh approximately 0.15 g of the dried product of this product, measure the amount of nitrogen by the Japanese Pharmacopoeia 13, General Test Method 31, and nitrogen determination method, and multiply this by 6.25 to obtain the amount of protein. Nitrogen measurement (mg) X6.25

Tank / amount of protein (%) = X100

Sampling volume (g) xiooo

6.25: Nitrogen protein conversion factor

From `` New Food Analysis Method '' edited by Japan Food Science Association

(2) Starch not detected

After adding 10 mL of water to 0.1 g of this product, heating and cooling, and adding 2 drops of iodine TS, it does not show a blue color.

(3) Fat 1.0% or less

Precisely weigh about 10 g of this product and put it in a cylindrical filter paper. Lightly fill the sample with the desulfurizer and place it in a Soxhlet extraction tube. Dry the flask of the receiver in an electric constant temperature drier at 100 to 105 ° C for 30 minutes to 1 hour, transfer to a desiccator, allow it to cool for 1 hour, weigh to 0. mg, and determine the constant weight. Add about 2/3 of getyl ether, connect the cooling pipe, and extract for 5 hours in an electric constant temperature water bath. After the extraction, remove the extraction tube, pull out the cylindrical filter paper with tweezers, connect it again to the cooling tube, heat it on a hot water bath, take out the flask when most of the ether in the flask has been transferred to the extraction tube, Warm still on the water bath and evaporate the rest of the ether. Wipe the outside of the flask with gauze or the like, put it in an electric constant temperature dryer at 100 to 105 ° C, dry it, transfer it to a desiccator, allow it to cool, and weigh it. W— Wo

Sample lipid (%): X 100

s

S: Sampling amount (g)

W: measured value (g)

W ₀ : Constant weight of receiver (g)

(4) Dietary fiber A: 80% or more B: 70% or more

Dry the product, weigh accurately about 1 g of it, add 50 mL of 0Ό8Μ phosphate buffer and 0.1 mL of heat-resistant amylase solution, and react in a boiling water bath for 30 minutes. After cooling to room temperature, adjust the pH to 7.5 ± 0.1 using a pH meter while adding about 10 mL of 0.275 M sodium hydroxide J-dimension. Add 0.1 mL of the protease solution and react in a water bath at 60 ° C for 30 minutes. After cooling to room temperature, adjust the pH to 4.5 ± 0.2 using a pH meter while adding about 10 mL of 0.325 M hydrochloric acid. To this, add 0.3 mL of Ami-mouth dalcosidase solution and react in a water bath at 60 ° C for 30 minutes. Add 200 mL of ethanol pre-heated to 60 ° C to this solution, and precipitate the dietary fiber for 1 hour at room temperature. This was filtered through a glass filter (G2) with a suction filter I, and the residue collected on the glass filter was filtered into 78 v / v% ethanol (20 ml_x 3 times) and 95 v / v% ethanol (10 mL 2 times). Then, wash sequentially with acetone (10 mL x 2). The residue is dried overnight at 105 ± 3 ° C with a glass filter, allowed to cool in a desiccator for about 1 hour, and weighed. Perform the above operation in a system that does not contain a sample to obtain a reagent blank value. R—B Dietary fiber (%) = x 100

w

R: remaining weight

B fiber blank

W: Sampling fee

(5) Calories

0.5kcal / g or less

Calculate by the following formula based on the measured values of protein and lipid (%) obtained above. Protein measured value X 3.47 + lipid measured value X 8.37 calories ikcal / g) =

100

* Energy conversion constant: From Alwater conversion coefficient

[Ash] Common to A and B 1.5% or less

Take 1 g of this product, and operate it according to JP13, general cloud it test method 16, and humility.

[Quantitative] Galactomannan A: 80% or more B: 60% or more (1) Reagents and reagents

■ β-mannanase (Japan / Quycon Inc.)

, -Galactosidase (Nippon Biocon Corporation)

-β-nicotinamide adenine dinucleotide (NAD) (Sigma) .Galactose dehydrogenase (Nippon Biocon Inc.)

■ 50mM acetate buffer (pH 4.5)

To 8 mL of sodium acetate TS, add 12 mL of dilute acetic acid and water to make 100 mL. ■ Tris buffer (pH8.6, 200mM)

Dissolve 2.42 g of 2-amino-2-hydroxyethyl-1,3-prononediol in 100 mL of water,

Add 0.1 .- hydrochloric acid to pH 8.6.

(2) Test method

1. Accurately weigh about 10 mg of this product, place it in a centrifuge tube, and add 5 mL of ethanol to 85-90. After incubating at C for 5 minutes, centrifuge (10 minutes, 3500 rpm / min). Carefully remove the supernatant. Repeat this twice.

2. Add 10.0mL of 50mM vinegar Mffi solution (pH4.5), stir, put in a boiling water tank, heat for 5 minutes, transfer to 40 ° C constant temperature water tank, and leave overnight (about 12 hours).

3. Add 20 µL of β-mannanase and incubate at 40 ° C for 180 minutes. Stir occasionally during that time

4 c

4. Centrifuge (3500 rpm / min, 10 minutes) and collect 0.1 mL of the supernatant. Add 0.2 mL of 50 mM acetate buffer (pH 4.5) and add 20 μL of galactosidase. Incubate the cells at 40 ° C for 60 minutes.

5. Add 2.5 mL of 200 mM Tris buffer (pH 8.6) to each cell, add 0.1 mL of NAD (0.1 g / 10 mL) and 8 L of galactose dehydrogenase, and incubate at 40 ° C for 60 minutes. 6. Measure the absorbance of this solution at 340 nm. <Calculation formula>

1

Galactose content (%) = (E-Eo) X X 1384.6

W

E: Absorbance at 340 nm (sample solution) E ₀ : Absorbance at 340 nm (blank) W: Sampling fee (mg)

1384.6: Conversion factor to galactomannan amount [Microbial limit test] A, B

1. Number of general viable bacteria 3 X 10 or less

2.Specific bacteria Escherichia coli: Not detected Perform the test by operating according to JP 13, general test method 41, and microorganism limit test method.

[Residual agricultural chemicals]

1. Endrin and dieldrin (including andrin): not detected

2. BHC: 0.2ppm or less

3. DDT: 0.2ppm or less (1) Equipment

Use gas chromatography with an electron capture detector (ECD).

(2) Standard product

■ — BHC standard product: Contains BHC 99% or more. Melting point: 157-159 ° C ■ β-BHC standard: β-BHC contains 98% or more. Melting point: 308-310 ° C'γ-BHC standard: r-BHC 99% or more. Melting point: 112-114 ° C ■ δ-BHC standard: <5- Contains 95% or more of BHC. Melting point: 137-140 ° C 'pp'-DDD Standard: Contains more than 98% of pp'-DDD. Melting point: 108-110. C ■ pp'-DDE standard products: Contains pp'-DDE99o / ₀ or more. Melting point: 88-90 ° C 'op'-DDT Standard: Contains more than 98% of op'-DDT. Melting point: 73-75 ° C ■ pp'-DDT standard: Contains 99% or more of pp'-DDT. Melting point: 102-104 ° C ■ Aldrin standard: Aldrin contains 97% or more. Melting point: 103-104 ° C. Contains endrin standard: 98% or more endrin. Decomposes at 200 ° C when determining melting point.

'Dildoline standard: Contains 98% or more dieldrin. Melting point is 177 ~ 179 ° C

(3) Preparation of sample solution

Add 10 g of powder, place in a flask, add 150 mL of benzene, and extract all day and night with stirring (about 12 hours or more). This is filtered and the filtrate is taken. Add an appropriate amount of anhydrous sodium sulfate, leave the mixture for 1 hour with occasional shaking, and filter into an evaporator flask. Then concentrate to exactly 20mL under reduced pressure I do.

Next, a column of 20 mm in inner diameter and 300 mm in length was charged with 20 g of synthetic magnesium gayate for column chromatography, and then about 8 g of anhydrous sodium sulfate suspended in hexane, and a small amount of hexane was placed at the top of the column. Allow hexane to drain to the extent that it remains. After injecting exactly 5 mL of the above concentrate into this column, inject hexane 'ether (17: 3), take about 300 mL of the first effluent, concentrate under reduced pressure to exactly 5 mL, and concentrate this sample. Make a solution.

(4) Test operation

1) Qualitative

'Column carrier: diatomaceous earth (177-250 m standard mesh sieve) was washed by refluxing with 6N hydrochloric acid for 2 hours, then washed with distilled water until the effluent became neutral, dried, and treated with methylsilazane (pyridine) Hexamethyldisilazane (special grade) immersed in trimethylchlorosilane (special grade) (5: 3: 1), washed with water for 10 minutes, and dried.

'Column packing: Incorporate 50/0 of gas chromatography silicon to the column carrier.

'Column: inner diameter 3mr, length 150-200cm, glass

'Column temperature: 180-210 ° C

'' Inlet temperature: 220—250 ° C

, Detector temperature: around 250 ° C

'Carrier gas: Use high-purity nitrogen. Aldrin flow rate in about 6 minutes Adjust to

2) Quantitation

Quantify by the peak height method or peak area method based on the test results obtained under appropriate conditions. If a large amount is detected, perform measurement using the internal standard method. When performing the internal standard method, use a standard product that has a retention time at a position where no peak is observed in the qualitative test as the internal standard product. ) Confirmation test (thin layer chromatography)

This test is performed only on test solutions whose 2) quantification result exceeds the pesticide residue standard based on the Food Sanitation Law.

The thin plate used in this test was prepared by spreading silica gel for thin layer chromatography containing 10% gypsum on a glass plate to a thickness of 500 m, heating at 130 ° C for about 1 hour, and activating it. I do. Store this in a container containing silica gel for drying and use it within one week.

Concentrate the sample solution in an appropriate amount (containing 10 to 20 Ug of organic chlorine agent) to several mL under reduced pressure, transfer to a small centrifuge tube, and send dry air or nitrogen to concentrate to about 0.1 mL. Take 10 to 20 L of the mixture with a micropipette or micro-syringe and attach it to a thin plate. Furthermore, apply 10 to 20 L each of 100 ppm standard and n-hexane to the above thin plate. Using n-hexane: ethyl acetate (9: 1) as an eluent, remove the thin plate when it rises about 10 cm by the ascending method and air dry it for about 5 minutes. Spray with 0.5% ortho-tolidine 'ethanol solution for about 5 minutes After leaving it for a while, irradiating it with UV light for about 1 day from a distance of several cm using a germicidal lamp (15W) produces brown-blue spots. Compare the spots of the sample solution and the standard sample. These standards and test methods shall be stipulated separately, and shall apply mutatis mutandis to the general rules of the Bureau and the General Test Methods of the Bureau.

“Fenugu Greek seed coat” standard and test method [Description] A brown powder with a peculiar smell and a bitter taste.

Saponin

Dissolve 1 g of this product in 20 mL of water and filter. The filtrate (1 L) is spotted on a silica gel thin layer plate, dried, and then developed with a developing solvent (cloth form: methanol: water = 65: 42.5: 10). After the thin plate was air-dried, a solution of antimony trichloride: 6N-hydrochloric acid = 1: 1 (w /) was sprayed, and 105. When incubated for 10 minutes in an electric furnace kept at C, a purple spot is obtained at Rf = 0.4-0.6. [Drying loss]

10.0% or less (105 ° C, 5 hours)

[Granularity]

70 mesh (212 m) or less (1) Protein 20% or more

The test method is the same as for the endosperm powder.

(2) Starch not detected

The test method is the same as for the endosperm powder.

(3) Fat 10% or less

The I test method is the same as that of the L-powder L powder.

(4) Dietary fiber 60% or more

The test method is the same as for the endosperm powder. (5) Calories

The test method is the same as for the endosperm powder.

[Residual agricultural chemicals]

1. Endrin and dieldrin (including andrin): not detected,

2. BHC: 0.2ppm or less 3. DDT: 0.2ppm or less

The test method is the same as for the endosperm powder.

[Microorganism limit test] 1. Number of general viable bacteria 2x10 ³ / g or less 2.Specific bacteria E. coli: not detected

Operate and test according to JP13, General Test Method 41, and Microorganism Limit Test Method. This standard and the H test method shall be stipulated separately, and shall apply mutatis mutandis to the general rules of the Bureau and the General Law of the Bureau. BRIEF DESCRIPTION OF THE FIGURES

Figure 1: Structural formula of fenuguricugalactomannan

Figure 2: Structural formulas of galactose and mannose in fenegriegalactomannan

Fig. 3: Formulation of froststanol-type steroidal saponins contained in Huene Greek seed coat

Fig. 4: Structural formula of 4-hydroxyisoleucine contained in Fenugreek seed coat Fig. 5: Flow chart for carrying out the invention

Figure 6: Stripper

Place the seeds of fenegriek or endosperm in a wire mesh with a diameter of 250 m (60 mesh), and rotate the propeller in the wire mesh with a motor. The seeds or endosperm create a force to invert each other, and make contact with the surrounding wire mesh, and the surface of the seed or endosperm is peeled off by friction and impact. The endosperm is hard and does not peel off much, but the seed coat is soft and peels off first. In the case of seeds, as the peeling of the seed coat progresses, the seed coat breaks due to impact, and the endosperm inside comes out. In the case of endosperm, the surrounding seed coat is peeled off, leaving only the endosperm. Figure 7: Air-flow crusher

This figure depicts the inside of an air-flow crusher, in which there are a plurality of spouted blades (rotating blades) in a container, which rotate at high speed. A jet stream is blown in from the outside, and a swirling vortex is generated inside. Fig. 8: Impact type crusher

This figure depicts the inside of an impact-type pulverizer, in which a hammer is mounted around a centered disk. The disk rotates at a high speed, and the hammer rotates at a high speed with a slight distance from the wall of the built-in container. Input: L Nugreek Endosperm is hit by a hammer and ground.

Figure 9: Mortar mill for sawing

This figure shows only the mortar used in the mortar mill, and shows a ring-shaped outer mill and a conical inner mill. The actual machine is installed inside ^! With a certain gap horizontally, with the inner mill entering the outer mill. The outer mill is fixed and only the inner mill rotates. The fenegriek seeds introduced from the top of the cone are crushed when passing through the gap and discharged through the groove to the bottom of the cone. BEST MODE FOR CARRYING OUT THE INVENTION

Separation of seed coat and endosperm

The best mode for carrying out the invention is shown as a flowchart in FIG. seed The seed coat and endosperm are separated from the mortar by passing them through a mortar-type crusher with a certain distance between the mortars, crushing the seeds, and passing the crushed seeds through an 18 to 30 mesh sieve. Separate by This process is repeated until only the endosperm remains.

Endosperm purification

The endosperm is crushed with an impact crusher. Since it is difficult to pulverize endosperm to 100 mesh or less with an impact-type powder mill, the seed coat that adheres to endosperm is pulverized to 100 mesh or less. This is removed using a sieve. This operation is repeated several times. The mesh size of the sieve and the number of revolutions of the crusher are adjusted according to the situation. When the endosperm that does not pass through the sieve is dissolved in water, the final product must be milky white and non-colored. The sieved product with a galactomannan content of 60-80% is commercialized as B-class endosperm powder. If there is no impact crusher, an air crusher can be used instead.

Endosperm crushing

Endosperm powder that has not passed through the 100 mesh is pulverized by an airflow pulverizer. After passing through a 70-mesh sieve, heat-sterilize and package.

Use of seed coat

When used as an extraction raw material.

Huene Greek oil

The seed coat is extracted with n-hexane. Any extractor may be used. A Soxhlet extractor that does not require a lot of solvent is economical. Filter the extract and remove the seed coat. The solvent is distilled off from the filtered extract to obtain n-greek oil. 4 /, Idoxyisoleucine

Dry sufficiently so that no n-hexane remains in the seed coat. Put the seed coat back into the extractor and extract with 70% ethanol solution. The ethanol concentration is adjusted as appropriate. The extraction with 70% ethanol solution is repeated at least twice. Filter the extract and remove the seed coat. The filtered extract is passed through a cation exchange resin column. Next, the ammonia solution is passed through the column to elute the adsorbed substances. The concentration of the ammonia solution is adjusted as appropriate. The eluate is concentrated under reduced pressure, and then concentrated and freeze-dried to obtain a powder containing crude 4 J, idroxyisoleucine. Package the powder.

Huene Greek Saponin

The effluent from the column is concentrated under reduced pressure. When the 95% ethanol solution is added to the concentrate, the saponin is screened. The precipitated saponin is filtered off. After washing the saponin with the 95% ethanol solution, vacuum-dry and pack the powder.

Fenugreek oleoresin

Concentration of the filtrate under reduced pressure leaves phenylene oleoresin. Pack it. Huene Greek protein

After the remaining seed coat is dried, it is extracted with alkaline water. Filter the extract and remove the seed coat of the residue. Neutralization of the extract with acid causes protein precipitation, and the protein precipitate is filtered off. The protein is dried under vacuum and packaged. When using the seed coat as a flavoring or health food ingredient.

Since the seed coat contains a large amount of oil, it hardens when powdered and then heat sterilized. Because it cannot be sterilized, heat sterilization before powdering, sterilize and pulverize. After crushing, immediately pack. Industrial potential

Since Huen Greek is used as a spice in curry, and most Japanese eat the curry, most Japanese are eating Fuen Greek, but most Japanese Not knowing about Huenegriek. In the United States and Canada, fenugreek is cultivated as livestock feed, but its seeds are not edible at all. Isn't fenigree grown for food in India and the Middle East? However, if you want to grow Huen Greek, you can do it anywhere in the world.

—On the other hand, the demand in Japan for galactomannan-based natural thickeners is about 3,000 tons per year. Until now, fenugreek galactomannan has not been used as a thickener, so these are all demands for guar gum and locust bean gum. Guar gum and Mouth cast bean gum rely on full imports due to limited production areas. Globally, demand for galactomannan is more than 10 times higher than in Japan. Since the physical properties of fenugreek gum are superior to guar gum and locust bean gum, it is highly likely that it will be used as a substitute for guar gum or locust bean gum. For example, if 10% of 3000 tons or 300 tons are used, 1200 tons of seeds will be required. (Because the endosperm is about 25% of the seed) New agriculture could occur in Japan to grow the fenegurieg Has the ability. This possibility is a common possibility not only in Japan but also in countries around the world.

Fenugreek gum has the characteristics of a functional food as well as the characteristics of a thickener. Therefore, another demand may arise. Then, if there is a great deal of demand in the future, this has the potential to become an industry. As demand for fenegriek gum increases, large amounts of seed coats are generated as by-products. The seed coat contains many active ingredients. By extracting this, it can be used as a raw material for medicines, cosmetics, flavors, and foods. The extracted seed coat is the same as cellulose, hemicellulose, lignin, etc., and the same as wood and wood. It also occurs in large quantities as biomass. If this is used as fuel for power generation, etc., a truly recycling-type environment can be created. By converting it to charcoal, it can be used as a raw material for generating hydrogen gas.

According to the present invention, it is not a dream to separate Huen Greek seeds into seed coat and endosperm to promote agriculture, develop various industries, treat diseases and obtain energy.

Claims

The scope of the claims

1. A method for physically separating fenegurik seeds into endosperm and seed coat (including cotyledons), purifying endosperm, and producing endosperm powder containing high-purity galactomannan.

2. Use of physically separated fenugreek seed coat (including cotyledons) and endosperm as various industrial raw materials including extraction