Process for separating pinitol from carob extract

Abstract

To provide a process for the separation of at least one inositol from a carob extract.SOLUTION: This process comprises the steps of: a) providing a filtered and demineralised carob extract having a Brix value greater than 60 and a pinitol content, in weight percent based on the weight of the extract, from 5 to 25%; b) subjecting the carob extract of step a) to a process of chromatographic separation of the pinitol, wherein the process comprises subjecting the extract to at least one passage through a chromatographic resin, thus obtaining an aqueous solution having a pinitol content, in weight percent based on the total weight of the solution, from 35 to 70%, and which has a Brix value of 20 or lower; and c) subjecting the aqueous solution thus obtained in step b) to a purification step, thus obtaining a purified aqueous solution having a pinitol content, in weight percent based on the total weight of the solution, of more than 55%.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates generally to the field of the food supplement industry, and in particular to pinitol-based food supplements. Regarding ment.

[0002] More particularly, the present invention relates to a process for separating pinitol from carob extract. Regarding. [Background technology]

[0003] Pinitol (3-O-methyl-1,2,4 cis-3,5,6 trans-hexa Hydroxycyclohexanol or 3-O-methyl-D-chiro-inositol) D-chiro-inositol (CH) with a molecular weight of 194.18 g / mol 14 O6) methyl ether It's Teru.

[0004] Pinitol (or D-pinitol) has the effect of lowering blood sugar levels and reducing insulin levels when administered orally. It is known for its effects on improving the function of the brain, as well as its use in the treatment of diabetes and obesity. Nitrite enhances creatine absorption, similar to when taken with carbohydrates. This allows you to get the amount of creatine you need without having to consume large amounts of carbohydrates. It becomes Noh.

[0005] Additionally, pinitol enhances muscle tissue function, increasing glycogen production in the muscles. It promotes glucose transport in muscle tissue. This activity of pinitol is beneficial for athletes It can be used to improve the performance of athletes. increases glucose uptake into muscle cells and replenishes glycogen stores in the muscles. This results in stable blood sugar levels and allows you to maintain high levels of energy for longer periods of time. You will be able to continue.

[0006] Pinitol is available in supplement form at a dose of 0.1 mg to 1.0 g per kg of body weight per day. It is administered orally in a non-oral or intravenous form or by incorporating it into food and drink. Administration is also possible.

[0007] Pinitol was first isolated from pine trees, but is also found in soybeans (weight percent based on the dry weight of soybeans). In some Asian countries where soy consumption is high, Pinitol intake from beans is estimated to be greater than 5 mg / kg / day.

[0008] Pinitol is a plant of Bougainvillea spectabilis and It is also found in Gliricidia sepium. It is also found in the fruit of the calab (Ceratonia siliqua), and Pinitol can be extracted using lithographic methods.

[0009] The carob tree is a long-lived, evergreen broadleaf fruit tree that grows slowly. Carob paste and seeds are used to make chocolate substitutes, while carob Many food thickeners and gelling agents are obtained from seed powder.

[0010] The composition of carob extract is typically expressed as a percentage by weight (based on the dry weight of the carob extract): Sucrose 40-65%, pinitol 7-15%, fructose 7-17%, glucose Therefore, carob contains a very high amount of pinitol. It is a rich source of pinitol, higher than, for example, soybeans and pine needles (0.5-1%). Includes.

[0011] Patent document 1 (Compania General del Algarrobo de España, SA) describes carob extract. A process for separating pinitol from extract is described. The sucrose contained in the syrup is converted to fructose and glucose, and the resulting syrup Specifically, the sugars contained in the syrup are separated by chromatography using a strong cationic resin. pinitol is separated from the solution to obtain an aqueous solution of pinitol with a purity of over 90%. The nitrol is separated.

[0012] Patent Document 2 (Amicogen Co. Ltd.) describes a method for separating pinitol from carob syrup. This method increases the pinitol content in the syrup and To obtain a product containing pinitol of low purity (40-50%), bacteria, yeast, or The process involves culturing the microbial cells or fungi. After separating the microbial cells, the resulting syrup is added with activated This process involves the treatment with charcoal and a crystallization process, which results in a purity of over 90%. A product containing nitrol is obtained.

[0013] All of the above processes can obtain pinitol with a purity of over 90%, but It is complicated and expensive. In fact, in the case of Patent Document 1, various filtration processes are envisaged, Then, the first desalting is carried out with a strong cation resin (Na), the extract is concentrated, and the cation resin The sucrose is converted into sucrose by passing it through an anion resin for desalting. Later, by chromatography by ISMB®, pinitol was chromatographed. Each pass requires the use of large amounts of water, and the output from each column Various steps are required to concentrate the resulting solution.

[0014] In the case of Patent Document 2, there is a process of culturing microorganisms in carob syrup, so A post-process to separate the cells of the food product is required. Both of these processes can be difficult to implement on an industrial scale, due in part to the need for additional equipment. do.

[0015] Pinitol can also be obtained by chemical synthesis, but this method is very expensive.

[0016] For this reason, in the field, pinitol from carob (and subsequently D-thiazol-2-one from pinitol) has been widely used. A simpler and cheaper process for isolating rho-inositol than the prior art processes It is required to provide [Prior art documents] [Patent documents]

[0017] [Patent Document 1] European Patent No. 1241155 [Patent Document 2] Korean Patent Publication No. 2004-0016338 Summary of the Invention [Problem to be solved by the invention]

[0018] The technical problem of the present invention is therefore to provide a method for producing carob extracts from carob, and more particularly from carob extracts. A practical, inexpensive, and versatile method for isolating pinitol (and D-chiro-inositol) The object of the present invention is to provide a process that is efficient, scalable, and high-yield. [Means for solving the problem]

[0019] This problem is solved by the present invention by isolating at least one inositol from carob extract. This process is resolved by: a) Brix value greater than 60 and pinitol content is 100% by weight based on the weight of the extract providing a filtered and desalted carob extract having 5-25% by weight of carob; b) subjecting the carob extract of step a) to chromatographic separation of pinitol; performing a process comprising applying said extract to a chromatography resin; At least one pass was performed to ensure that the pinitol content was a weight percentage based on the total weight of the solution. obtaining an aqueous solution having a Brix value of 20 or less and a concentration of 35-70% and, c) Purifying the aqueous solution obtained in step b) to obtain a pinitol-containing solution. obtaining a purified aqueous solution having a content of greater than 55% by weight based on the total weight of the solution; It has.

[0020] The term "pinitol" in this patent refers to pinitol in the D configuration (D-pinitol). D-pinitol is the only configuration of pinitol present in carob extract. do.

[0021] As used herein, "carob extract" refers to the maceration of chopped carob pods. This refers to an aqueous solution obtained by separating a coarse solid residue from the aqueous solution obtained by squeezing a liquid containing lactic acid bacteria.

[0022] Preferably, the at least one inositol is pinitol and / or D-thiamine. It is preferably selected from rho-inositol, more preferably pinitol.

[0023] Soaking is generally performed by mixing the pods with water in a weight ratio of about 1:3 and soaking them at a temperature of 60 to 90°C for 1 to 2 hours. The process is carried out for 24 hours at a pH of 4.5 to 5.5. The pressing is generally carried out by a press, e.g. It is carried out continuously.

[0024] The resulting solution is generally dark in color and contains suspended particles. Generally glucose, fructose, sucrose, pinitol, and other sugars or impurities It contains malt and typically has a Brix value of 10-30.

[0025] The filtered extract of step a) may be filtered using any filtration technique known in the art, preferably a rotary vacuum filter. More preferably, the filter aid comprises perlite.

[0026] Preferably, the distribution of particle diameters of perlite greater than 160 μm is 5% (w / w) to 10% (w / w), more preferably 7% (w / w).

[0027] Preferably, the density value of the perlite is between 90 and 130 g / l, more preferably 110 g / l. is.

[0028] Perlite suitable for the purposes of the present invention is, for example, Randalite® W24 ( The French company is the Ceca Arkema Group.

[0029] Perlite is a soft rock composed of aluminum silicate that expands when heated. This expanded material is then ground to produce filter aids of various sizes (degrees).

[0030] Preferably, in addition to or instead of filtration by a rotary filter, a filter effluent is used. A bell filter (also called a precoat filter) with vertically arranged elements This may include a filtration step.

[0031] Preferably, the filter medium comprises diatomaceous earth.

[0032] Preferably, the diatomaceous earth comprises SiO2.

[0033] Preferably, the diatomaceous earth is of the flux-calcined type.

[0034] A suitable filter medium for the purposes of the present invention is, for example, Dicalite Speedplus® trademark) or Dicalite 6000 (Palumbo Trading, Srl, Italy).

[0035] Preferably, the filtration is performed using a rotary filter and / or a bell filter. Additionally or alternatively, filtration may be performed using tangential filtration techniques known in the art. The method may include a filtration step by passing the mixture through a tangential filter. The filter is equipped with a pore size of 0.45 μm or less.

[0036] Preferably, the carob extract of step a) is concentrated. The concentration can be carried out by any of the methods known in the art. Methods such as thermal concentration, preferably at a temperature of 40 to 90°C, 6000 to 10000 l / h This is done at a flow rate of .

[0037] Preferably, the carob extract of step a) is decolorized.

[0038] Preferably, the decolorization is carried out by adsorption chromatography.

[0039] Preferably, the decolorization is performed by treating the carob extract with an adsorbent resin, more preferably styrene-divinyl by passing it through an adsorbent resin containing a diphenyl benzene (DVB) copolymer-based matrix. Selection of the appropriate resin and process parameters is within the skill of one of ordinary skill in the art. Resins suitable for decolorization are available from Sepa, Resindion Srl (Milan, Italy). beads® SP207 adsorbent resin.

[0040] Preferably, the carob extract of step a) is purified by cation exchange chromatography and anion exchange chromatography. It is desalted (or rectified) by ion exchange chromatography.

[0041] Preferably, the carob extract in step a) is prepared by subjecting the carob extract to at least one anion. anion exchange resin and at least one cation exchange resin, more preferably a weak anion exchange resin It is desalted (or conditioned) by passage through a resin and a strong cation exchange resin.

[0042] Preferably, the carob extract in step a) is prepared by subjecting the carob extract to at least one anion. Anion exchange resin, more preferably a weak anion exchange resin, followed by a cation exchange resin, more preferably a weak anion exchange resin. The solution is desalted by passing it through a strong cation exchange resin.

[0043] Preferably, the carob extract in step a) is subjected to at least two weakly acidic conditions. Desalted by passing through an ion exchange resin and at least two strong cation exchange resins. do.

[0044] Preferably, the carob extract in step a) is purified by subjecting the carob extract to two weak anion exchangers. The solution is desalted by passage through a resin and two strong cation exchange resins.

[0045] Preferably, the carob extract is passed at least once through a weak anion exchange resin, followed by a strong cation exchange resin. Before passing it through the ion exchange resin, the carob extract is passed through a strong anion exchange resin.

[0046] Preferably, said at least one passage of the carob extract through a weak anion exchange resin comprises: This is the final pass.

[0047] In a preferred embodiment, the desalting comprises treating the carob extract with: i. passing the carob extract through a weak anion exchange resin for the first time; ii. passing the carob extract through a strong cation exchange resin for the first time; iii. passing the carob extract through a weak anion exchange resin for a second time; iv. passing the carob extract through a strong anion exchange resin; v. Passing the carob extract through a strong cation exchange resin for the second time. This is how it is done.

[0048] As is known, strong ion exchange resins can operate over a pH range of 0 to 12, Weak ion exchange resins are only capable of exchange to a narrower extent. Weak cation exchange resins are weak anion exchange resins act in the neutral range, while weak anion exchange resins act in the basic range.

[0049] A weak anion exchange resin suitable for the present invention is Relite RAM1 (registered trademark) Resindion Srl, Milan, Italy), Dowex® MWA-1 (Dowex, Japan) Chemical Company), Purolite(trademark) A100 (Dow Chemical Company, Japan) ·Company) etc.

[0050] Strong anion exchange resins suitable for the present invention include Relite RAP1 (registered trademark) Resindion Srl, Milan, Italy; Amberlite® IRA900 (Ola Lenntech BV (Netherlands), Purolite® A500 (Lenntech BV (Netherlands) ) are mentioned.

[0051] Strong cation exchange resins suitable for the present invention include Relite RPS (registered trademark) Resindion Srl, Milan, Italy; Amberlite® IRC200, Lenntech BV (Denmark), Purolite® A150 (Lenntech BV (Netherlands) Examples include:

[0052] Preferably, the desalting step is carried out continuously, i.e. without interrupting the desalting process. can be.

[0053] Preferably, the desalting process removes 100% of the impurities and ions present in the extract.

[0054] Preferably, the pH of the solution leaving the desalting step is between 3 and 5. In practice, within this pH range This can prevent the extract from browning.

[0055] Selection of appropriate resins and process parameters is within the skill of one in the art.

[0056] Preferably, the carob extract of step a) has a Brix value of at least 65.

[0057] Brix (Bx) is the percentage (%w / w) of solid matter dissolved in a liquid. In the present invention, the Brix degree is measured by one of the methods known in the art. Thus, for example, it can be carried out using a refractometer. A refractometer suitable for the purposes of the present invention is This is the Atago RX-9000CX model (Atago USA, Inc.).

[0058] Preferably, the conductivity value of the extract of step a) is between 70 and 110 μS / cm, more preferably The conductivity is 90 to 100 μS / cm. The conductivity is measured according to a method known in the art, for example, For example, this can be done using a conductivity meter.

[0059] Preferably, the pH of the carob extract in step a) is between 2 and 4.5, more preferably 2.5. ~3.5.

[0060] Preferably, the absorbance value of the carob extract in step a) is measured using a quartz cuvette with a 1 cm path length. , measured at 430 nm, is 0.005 to 0.030, more preferably 0.010 to 0. The number is 020.

[0061] Preferably, the carob extract of step a) comprises 5% by weight based on the weight of the extract. It contains up to 20%, more preferably 10-15% pinitol.

[0062] Preferably, the carob extract of step a) comprises 5% by weight based on the weight of the extract. Contains ~15%, more preferably 8-10% sucrose.

[0063] Preferably, the carob extract of step a) comprises, in weight percentages based on the weight of the extract: 5-15%, more preferably 8-10% sucrose; 5-20%, more preferably 10 ~15% pinitol; 20-50%, more preferably 30-40% fructose; It contains 0 to 50%, more preferably 30 to 40% glucose.

[0064] Preferably, step b) comprises treating the carob extract of step a) with, for example, Diaion™ Strong cation exchange resin (Na ) such as UBK530 resin (Resindion Srl, Milan, Italy) + Other resins suitable for the purposes of the present invention include Diai on™ UBK535, UBK550, and UBK555 (Resi, Milan, Italy) Examples of resins include those of Andion Srl.

[0065] Preferably, step b) is carried out by (continuous) simulated moving bed chromatography (SMB chromatography). more preferably by improved (continuous) chromatographic separation ("improved Simulated Moving Bed (ISMB)), such as ISMB® (Improved Simulated Moving Bed, Mitsubishi Chemical This is carried out by the Japan Industrial Co., Ltd.

[0066] Preferably, the aforementioned simulated moving bed chromatography method (SMB chromatography), Preferably, the aforementioned continuous chromatographic separation ISMB, in particular ISMB®, is It is carried out using four columns.

[0067] As is known, simulated moving bed chromatography is a continuous multi-column chromatograph. The Fee process is a technology known since 1961 that continuously processes a purified binary mixture. It is used when preparing

[0068] The aforementioned ISMB method, developed by Mitsubishi Chemical Corporation (Tokyo, Japan), is an improvement of the SMB method. This allows the two components to be separated.

[0069] Preferably, in step b), elution is carried out with demineralized water.

[0070] Preferably, the aqueous solution obtained in step b) has a Brix value of 15 or less, more preferably 1 is less than or equal to 0.

[0071] Preferably, the pinitol content of the aqueous solution obtained in step b) is 0.01g based on the total weight of the solution. The weight percentage based on the total weight is 50 to 70%, and more preferably 60 to 70%.

[0072] Preferably, the sucrose content of the aqueous solution obtained in step b) is The weight percentage is 2 to 8%.

[0073] Preferably, the aqueous solution obtained in step b) has a weight percentage based on the total weight of the solution , sucrose content 2-8%, glucose content 20-32%, pinitol content The content of fructose is 50 to 70%, more preferably 60 to 70%, and the fructose content is 0 to 6%.

[0074] Preferably, at the end of step b), a second solution (wastewater) having a Brix value of 25 to 40 is obtained. is also obtained.

[0075] Preferably, the pinitol content of the second solution is a percentage by weight based on the total weight of the solution. In most cases, it is less than 10%.

[0076] Preferably, this second solution contains, by weight percent based on the total weight of the solution, sucrose Content 0-4%, glucose content 2-10%, pinitol content 2-7%, full The lactose content is 85-95%.

[0077] Preferably, the purification step c) comprises concentrating the solution obtained in step b), preferably by heating. This includes the step of:

[0078] Preferably, the step of concentrating the solution by heat is carried out to a Brix value of 60 or more, more preferably The solution is heated to a temperature of 25 to 60°C until the temperature reaches 70 or more, more preferably 70 to 75°C. This includes heating to

[0079] Preferably, in step c), the concentration of the solution is followed by a crystallization step of the resulting solution. cormorant.

[0080] Preferably, the crystallization step is carried out by stirring the concentrated solution for 18 to 20 minutes until crystals form and precipitate. This is done by keeping the temperature at 25°C for 3 to 10 days.

[0081] Preferably, in step c), the concentrate obtained is etched with ethyl acetate until pure crystals are formed. After adding ethyl alcohol (e.g., 71% by volume of ethyl alcohol solution), the crystals are precipitated. This completes the crystallization.

[0082] The parameters and materials used in the purification step c) can be varied to obtain the desired results. is within the capabilities of one skilled in the art.

[0083] Preferably, at the end of purification step c), at least 70%, more preferably at least 8 0%, more preferably at least 85%, even more preferably at least 90%, most preferably Preferably, a concentrate containing pinitol in a purity of at least 95% is obtained.

[0084] In this patent application, the purity of a component is based on the weight of the solution or crystals containing that component. It is understood that the weight percentages are expressed as weight percent of the component based on the weight of the component.

[0085] Preferably, the concentrate obtained in step c) containing pinitol at a purity of more than 55% is centrifuged. A precipitate containing pinitol and a supernatant containing glucose are obtained.

[0086] Preferably, the resulting precipitate is heated, more preferably at about 45° C. for at least 2 days. By dehumidifying, pinitol is obtained in the form of a white powder with a purity of at least 95%.

[0087] Preferably, after step c), the aqueous solution obtained in step c) or the precipitate containing pinitol is Pinitol obtained after dehumidification of the sediment was subjected to acid hydrolysis to obtain D-tyrofoam. Then, an aqueous solution containing D-chiro-inositol was obtained, and the aqueous solution containing D-chiro-inositol was then subjected to strong anionic An aqueous solution containing D-chiro-inositol is prepared by passing it through an ion exchange resin at least once. Preferably, D-chiro-inositol is obtained by chromatographic separation from D - containing chiro-inositol in a weight percentage of at least 95% based on the total weight of the solution , preferably a step d) is carried out to obtain an aqueous solution having a Brix value of 1 or less.

[0088] Preferably, in step d), the acid hydrolysis is carried out by adding an aqueous solution of pinitol (e.g., 33% This is done by adding HCl (v / v).

[0089] Preferably, following the addition of HCl, the resulting aqueous solution is allowed to stand for at least 12 hours, more preferably or a boiling process for at least 24 hours.

[0090] Preferably, in step d), the strong anion exchange resin is RAP1® (Italian Resindion Srl (Milan, Italy), Amberlite IRA900 (Netherlands) Lenntech BV (Netherlands), and Purolite® A500 (Lenntech BV (Netherlands) BV), preferably RAP1®.

[0091] Preferably, in step d), the aqueous solution is dehydrated before passing it through the strong anion exchange resin. The color step is more preferably carried out by adding activated carbon to the solution.

[0092] Preferably, the activated carbon is present in an amount of 50 to 150 g per hectoliter of solution, more preferably It is added to the solution at a concentration of 80-120 g per hectoliter of solution.

[0093] The activated carbon is selected from activated carbons with a median diameter of 4 to 50 μm, more preferably 8 to 15 μm. It is preferable to do so.

[0094] Preferably, the BET of the activated carbon is 1200 to 2000 m 2 / g, more preferably 15 00~1800m 2 / g.

[0095] Activated carbon suitable for the purposes of the present invention is, for example, Picapure HP 120 (Italian Pica Italia SpA (Italy) and Decoran (registered trademark) (AEB (Italy) )) are mentioned.

[0096] Median diameter (MT50 or d50) is understood to be measured using a laser particle meter. 50 weight percent of the particles have a small diameter and 50 weight percent of the particles have a large diameter. The diameter corresponds to the particle size measured by the laser particle meter as described above. It means IZ.

[0097] BET surface area is measured according to the procedure of ASTM D-3037 / 89. It is intended to be what is intended.

[0098] Preferably, in step d), the Brix of the aqueous solution introduced into the strong anion exchange resin is The value is 6.5 or higher.

[0099] Preferably, in step d), the pH value of the aqueous solution leaving the strong anion exchange resin is adjusted to a basic The pH value is preferably 8 to 12.

[0100] Preferably, the D-chiro-inositol obtained in step d) that comes off the strong anion exchange resin is By acidifying the aqueous solution containing the compound, the pH of the acidified aqueous solution is 3 to 5, preferably about 4. A solution is obtained.

[0101] Preferably, the acidification step is carried out with a weak acid, such as citric acid.

[0102] Preferably, the aqueous solution containing D-chiro-inositol obtained in step d) is concentrated to obtain and a Brix value of 60 or more, more preferably 65 or more, and even more preferably 70 or more. A condensed solution is obtained.

[0103] Preferably, the resulting concentrated aqueous solution is subjected to crystallization, more preferably, the concentrated aqueous solution is subjected to crystallization at about Keep at a temperature of 7-10°C for 2-6 hours.

[0104] Preferably, after crystallization, the D-chiro-inositol is subjected to a dehumidifying treatment, more preferably an absorption treatment. By carrying out the above steps, the purified product is at least 90%, more preferably at least 95% purified. D-chiro-inositol is obtained.

[0105] Preferably, the yield of pinitol is based on the weight of the carob pods from which the pinitol was extracted. At least 3% by weight, more preferably at least 5%, and even more preferably is at least 7%, most preferably 7 to 10%.

[0106] Preferably, the yield of pinitol is based on the weight of the carob pods from which the pinitol was extracted. <15% by weight or less.

[0107] Preferably, the yield of pinitol is determined based on the weight of the starting pinitol (contained in the pods). The weight percentage based on the amount is 80% or more.

[0108] Preferably, the process of the present invention is carried out continuously.

[0109] Therefore, the process of the present invention is useful for the separation of pinitol, the separation of D-chiro-inositol, In fact, by carrying out the process up to step c), By continuing the process, pinitol can be converted to D-chiro-inositide. It is also possible to obtain D-chiro-inositol using a portion of pinitol. By obtaining pinitol, it is possible to obtain both pinitol and D-chiro-inositol. be.

[0110] Surprisingly, the process of the present invention allows for greater production of pinitol and and / or D-chiro-inositol in a simpler, faster, cheaper and more pure form. It was discovered that it was possible to obtain

[0111] In fact, the process of the present invention contemplates a relatively small number of passes compared to the prior art. .

[0112] Furthermore, the presence of a desalting step, especially when carried out according to a preferred embodiment of the present invention, In this case, pinitol is separated from a relatively high concentration aqueous solution containing pinitol at a relatively high concentration. This allows the aqueous solution to be diluted (if the pinitol content is the same). Since the amount required is relatively small, it is possible to further streamline the process. The amount of water required for dilution each time the sample is passed through the chromatography is relatively small, reducing costs and It also leads to a reduction in waste.

[0113] In a preferred embodiment, the desalting step uses a specific resin sequence, in particular: i) weak anion resin, ii) strong cation resin, iii) weak anion resin, iv) strong anion resin, and finally v) strong The preferred sequences i to v, which comprise the cationic resins in this order, are particularly advantageous.

[0114] Anion and cation resins are used alternately, as well as strong and weak ion exchange resins. By alternating with ion exchange resin, a particularly high recovery rate of pinitol can be achieved. In addition, by passing the solution through a strong cation resin at the end, the pH of the discharged solution becomes acidic. This is particularly advantageous because browning of the solution can be avoided and a separate decolorization step is not required.

[0115] A further advantage of the method of the present invention is that it can be carried out continuously. It allows for greater simplification and automation compared to continuous processes, resulting in increased speed. [Brief explanation of the drawings]

[0116] [Figure 1]FIG. 1 is a block diagram showing a preferred embodiment of a portion of the process of the present invention, leading to obtaining an aqueous solution of step b) from carob pods, having a pinitol content of 35-70% by weight based on the total weight of the solution, and a Brix value of 20 or less. [Figure 2] 1 shows the results of HPLC analysis related to the determination of the composition of the steeped and pressed carob extract described in Example 1. [Figure 3] FIG. 1 is a diagram of the passes associated with the desalting step according to a preferred embodiment (Example 1) of the present invention. [Figure 4] 1 shows the results of HPLC analysis related to determining the composition of carob extract after filtration, bleaching, conditioning (demineralization), and concentration as described in Example 1. [Figure 5] FIG. 1 is a block diagram of a preferred embodiment of the process of the present invention, showing the process from step b) of obtaining a purified aqueous solution in step c) from an aqueous solution having a pinitol content of 35-70% by weight based on the total weight of the solution and a Brix value of 20 or less (Example 1). [Figure 6] 1 shows the results of an HPLC analysis relating to the determination of the composition of the aqueous solution obtained in step b), described as fraction 1 of Example 1, having a pinitol content of 35-70% and a Brix value of 20 or less. [Figure 7] 1 shows the results of HPLC analysis related to the determination of fraction 2 described in Example 1. [Figure 8] 1 shows the results of HPLC analysis relating to the determination of the composition of the purified aqueous solution obtained in step c) described in Example 1, having a pinitol content of greater than 55% by weight based on the total weight of the solution. [Figure 9] FIG. 1 is a block diagram showing a preferred embodiment of a part of the process of the present invention, showing the process from the purified aqueous solution of step c) to obtaining an aqueous solution of step d) containing D-chiro-inositol and having a Brix value of 1 or less (Example 2). [Figure 10] 4 shows the results of an HPLC analysis relating to the determination of the composition of the aqueous solution of step d) containing D-chiro-inositol of Example 2 and having a Brix value of ≦1. DETAILED DESCRIPTION OF THE INVENTION

[0117] The present invention will now be described with reference to example embodiments, which are provided for purposes of illustration and not limitation. will be further explained.

[0118] Example 1 Pinitol separation process (Figures 1 to 8)

[0119] Chop 500 kg of carob pods into pieces of about 1 cm and mix in a ratio of 1 part pods to 3 parts water. The pieces were then pressed to determine the composition (dry weight of juice). (based on dry weight percentage) sucrose 62.5%, glucose 11.2%, pini The composition of the product is 10.1% ethanol, 16.1% fructose, and 0.5% impurities (composition in Figure 2). Salobrant extract was obtained.

[0120] The above composition was determined by HPLC using HO as the eluent, a flow rate of 0.6 ml / min, and a column temperature of 7 5°C, column size 8mm I.D., column length 300mm, functional group Ca, cation exchange resin It was determined using fat.

[0121] The Brix value of the resulting extract was 18.

[0122] The extract was filtered through a rotary filter under vacuum using Perlite Randal ite® W24 (Ceca Arkema Group, France) was used as a filter aid. The mixture was filtered.

[0123] The filtrate is then filtered through a vertically arranged filter element containing diatomaceous earth, specifically D icalite Speedplus (Palumbo Trading Srl, Italy) as a supporting material It was filtered a second time through the used Bell filter.

[0124] The filtrate was then filtered through a Tangier filter using a membrane with a pore size of approximately 0.45 μm as the filter element. The mixture was filtered a third time through a potential filter.

[0125] The filtered extract was purified by Sepabeads® SP207 (Resindio, Italy). n Srl) adsorption resin for decolorization, then desalting (or passing through the following resins in the following order): was adjusted) (see diagram in Figure 3). 1) Column 1: Relite RAM1 / M (Resindion Srl, Milan, Italy) (weak anion) 2) Column 2: Relite RPS (Resindion Srl, Milan, Italy) (Strong Yang Io hmm) 3) Column 3: Relite RAM1 / M (Resindion Srl, Milan, Italy) (weak anion) 4) Column 3: Relite RAP1 (Resindion Srl, Milan, Italy) (strong negative on) 5) Column 4: Relite RPS (Resindion Srl, Milan, Italy) (Strong Yang Io hmm)

[0126] Table 1 shows the properties of each resin.

[0127] [Table 1]

[0128] The operating conditions for each column are shown in Table 2.

[0129] [Table 2]

[0130] The properties of the four resins mentioned above are shown in Table 3. [Table 3]

[0131] The operating conditions for the four resins mentioned above are shown in Table 4.

[0132] [Table 4]

[0133] The conductivity of the extract was 100 μS / cm, the pH was 3.10, and the color was measured as The absorbance (Abs) was measured at 430°C in a quartz cuvette with a path length of 1 cm. value).

[0134] Next, the obtained extract is heated and passed through a vacuum condition from an inlet at a temperature of 80°C to an inlet at a temperature of 45°C. After passing it through the outlet and concentrating it, 65°Bx was obtained. The composition of this extract was (percent dry weight based on dry weight) sucrose 5%, glucose 38%, pinito The ethanol content was 15%, fructose 38%, and impurities 4% (see Figure 4).

[0135] The above composition was analyzed by HPLC as described above, with eluent HO and a flow rate of 0.6 ml / min. , Column temperature 75℃, Column size 8mm I.D., Column length 300mm, Functional group Ca, It was determined using a cation exchange resin.

[0136] The resulting concentrated extract was then passed through four UBK 530 columns (Resinde, Milan, Italy). ion srl) and an ISMB (registered trademark) plant (Improved Simulated Moving Bed, Mitsubishi Chemical Industries, Ltd. The solution was supplied to a supplier (company) and eluted with demineralized water.

[0137] Other operating parameters are shown in Table 5

[0138] [Table 5]

[0139] Legend: W: Water flow velocity F: Feeding speed P: Volume of purified solution R: Volume of concentrate ("waste")

[0140] Table 6 shows the ISBM operating conditions.

[0141] [Table 6]

[0142] This chromatography yielded two liquid fractions with the compositions shown in Table 7 (respectively See Figures 6 and 7).

[0143] [Table 7]

[0144] The above compositions were determined by HPLC as previously described.

[0145] Next, fraction 1 containing 70% pinitol was separated from the inlet at 80°C under vacuum and transferred to the inlet at 4°C. The mixture was concentrated by heating until the Brix value reached 73, and then refrigerated for 2 days. The mixture was kept at 0°C to form pinitol crystals.

[0146] After crystals have formed and settled, add 71% by volume of ethyl alcohol to 5 parts of the concentrated solution. The crystals were purified by adding 2 parts of coal to the concentrate to obtain pinitol with a purity of over 95%. The composition of the crystals obtained was (weight percent based on the weight of the concentrate): 3% glucose, 1% pinyl alcohol, 1% ethanol. The total content was 96.5% tallow, 0% sucrose, and 0% fructose (see Figure 8).

[0147] The above composition was determined by HPLC under the conditions described above.

[0148] This purified solution was centrifuged at 4000 rpm to remove pinitol and alcohol. A precipitate and a supernatant containing glucose and alcohol were obtained.

[0149] The precipitate was heated at 45°C for 2 days to remove moisture, and the purity of pinitol was over 95%. 30 g of powder was obtained.

[0150] This result indicates that 90 weight percent of the weight of pinitol contained in the starting pods is This corresponds to the yield of pinitol obtained in the same manner as above.

[0151] The resulting white powder sample was then compared with a standard pinitol sample to confirm that the substance was pinitol. It was confirmed that

[0152] For this purpose, an equal aliquot of each sample was mixed with MeOH / H2O in an 80 / 20 ratio. The samples were dissolved in the mixture to a concentration of 15 ppm (μg / ml).

[0153] The samples were analyzed by LC / MS (liquid crystallography) using a Luna NH2 column (150 x 2.2, 3 μm). The analysis was carried out using a solvent-based chromatography / mass spectrometry (PELC). The elution was carried out by isocratic elution using a mobile phase consisting of water (20%) and ethanol (20%). The analytical procedure was carried out for 15 minutes, and the flow rate used was 300 μl / min.

[0154] Equipment used: Water Micromass Q-TOF Premier mass spectrometer .

[0155] Analysis confirmed the concordance of the two samples.

[0156] Example 2 D-chiro-inositol isolation process (Figures 9 and 10) 30 g of powdered pinitol with a purity of over 95% obtained in Example 1 was added to a 1-liter flask. The solution was added to 16 g of water and 104 g of 33% hydrochloric acid was added.

[0157] The solution was heated (45°C to 60°C) for 20 minutes, and 40 ml of 7.2 N hydrochloric acid was added. 50 ml of water was added under reflux, and the solution was then boiled and stirred for 24 hours at constant reflux. Boiling continued for a while.

[0158] After 24 hours, the solution was stirred for 60 minutes, and activated carbon was added to the solution (100-150 g / h) to obtain 1160 ml of a solution with a Brix value of 6.5.

[0159] The solution was then filtered to remove the brown component that formed during heating. Dicalite Speedplus (registered trademark) (Palumbo Trading, Srl, Italy) ) diatomaceous earth and 50 weight percent Randalite® W24 as an auxiliary agent (Ceca Arkema Group, France) Rotary milling under vacuum using a mixture of perlite Filtration was carried out using a filter.

[0160] At this stage, the solution should have a pH of 1 and a clarity of 2 NTU (Nephelometric Turbidity Units). and was colorless.

[0161] The solution was then neutralized.

[0162] The solution was then passed through a strong anion exchange resin (Relite RAP1) to adjust the pH to 9. After reaching pH 10, the solution was acidified with citric acid to a pH of 4.0.

[0163] The resulting solution had a Brix value of 0.3 and was subsequently concentrated to a Brix value of 70. It shrunk.

[0164] The solution was then kept at a temperature of 8°C for 24 hours to allow the crystallization of D-chiro-inositol. It was.

[0165] The D-chiro-inositol content of this concentrate was 95% or more.

[0166] Finally, the concentrate is dehumidified by absorption to produce a white powder of D-chiro-inositol with a purity of over 95%. The final yield was 29g.

[0167] This result corresponds to a yield of nearly 100%.

[0168] Next, the obtained white powder sample and D-chiro-inositol were analyzed using the method described in Example 1 above. Comparison with a standard sample of D-chiro-inositol confirmed that the substance was D-chiro-inositol.

[0169] Analysis confirmed the concordance of the two samples.

Claims

1. 1. A process for isolating at least one inositol from carob extract, comprising: a) providing a filtered and desalted carob extract having a Brix value greater than 60 and a pinitol content of 5-25% by weight based on the weight of the extract; b) subjecting the carob extract of step a) to a chromatographic separation process for pinitol, which comprises passing the extract through a chromatographic resin at least once to obtain an aqueous solution having a pinitol content of 35-70% by weight based on the total weight of the solution and a Brix value of 20 or less; and wherein the carob extract of step a) is desalted by passing it through at least two weak anion exchange resins and at least two strong cation exchange resins; After passing the carob extract through a weak anion exchange resin at least once, the carob extract is passed through a strong anion exchange resin before passing it through a strong cation exchange resin.

2. 10. The process of claim 1, further comprising: in step a), the carob extract is decolorized.

3. The desalting step comprises: i. passing the carob extract through a weak anion exchange resin for the first time; ii. Passing the carob extract through a strong cation exchange resin for the first time; iii) passing the carob extract through a weak anion exchange resin for a second time; iv. passing the carob extract through a strong anion exchange resin; v. passing the carob extract a second time through a strong cation exchange resin.

4. 4. The process of claim 1, wherein the carob extract of step a) comprises 5 to 20%, or 10 to 15%, pinitol, in weight percentages based on the weight of the extract.

5. 5. The process according to any one of claims 1 to 4, wherein step b) is carried out by the "Simulated Moving Bed Chromatography (SMB Chromatography)" method, by Improved Continuous Chromatographic Separation (ISMB), or by ISMB® (Mitsubishi Chemical Industries, Ltd.).

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