METHOD FOR OBTAINING A BOTANICAL EXTRACT

Abstract

A method for preparing a botanical extract includes extracting plant matter with ultrapure water, and separating the solid plant matter from the liquid fraction, concentrating the liquid fraction, and purifying the concentrated liquid fraction by tangential flow filtration.

Claims

1. A method for preparing a botanical extract, comprising: a) extracting a plant matter with ultrapure water to obtain a liquid fraction and solid plant matter, and separating the solid plant matter from the liquid fraction; b) concentrating the liquid fraction; and c) purifying the liquid fraction obtained after concentrating by tangential flow filtration.

2. The method according to claim 1, wherein the plant matter is selected from the group formed by pomegranate, cucumber, broccoli, strawberry, Epilobium angustifolium, blueberry, grape or red vine, camomile, maypop, parsley, sage, arnica, mint, citrus, orange, lemon, grapefruit, cinnamon, Serenoa repens, nettle, sunflower, spinach, olive leaf, magnolia, bearberry, grape seed, olive, cannabis, coffee beans, artichoke flower, yerba mate, cocoa, Thea sinensis, ginseng, horse chestnut, turmeric, pine bark, artichoke plant, echinacea, ginseng, persimmon, pear, broccoli.

3. The method according to claim 1, wherein the extract obtained contains the active substances selected from the group formed by alkaloids, ellagitannins, anthocyanins, flavones and flavonols, flavonoids, sterols, ecdysteroids, steroids, iridoids, lignans, phenols, curcuminoids, cannabinoids, phenylpropanoids, saponins, oligomeric procyanidins, isothiocyanates, imino and amino sugars.

4. The method according to claim 1, wherein said active substance is not abscisic acid.

5. The method according to claim 1, wherein the plant matter is dried.

6. The method according to claim 1, wherein a volume of solvent equivalent to between 3 and 6 times the weight of the plant matter to be extracted is used.

7. The method according to claim 1, wherein the extraction is carried out at a temperature of 50° C. to 75° C.

8. The method according to claim 1, wherein the size of the membrane in the tangential flow filtration is between 100 Da and 100,000 Da.

9. The method according to claim 1, wherein the pressure in the tangential flow filtration is between 4 and 30 bar.

10. A composition comprising the botanical extract obtained according to claim 1 and at least one additional component.

11. The composition according to claim 10, wherein the extract obtained contains the active substances selected from the group formed by alkaloids, ellagitannins, anthocyanins, flavones and flavonols, flavonoids, sterols, ecdysteroids, steroids, iridoids, lignans, phenols, curcuminoids, cannabinoids, phenylpropanoids, saponins, oligomeric procyanidins, isothiocyanates, imino and amino sugars.

12. The composition according to claim 10, wherein the additional component is selected from the group formed by colloidal silica, talc, tricalcium phosphate, magnesium stearate, carbohydrates, polysaccharides, maltodextrins, cyclodextrins, water, polyols, cellulose, starches, and mixtures thereof.

13. The composition according to claim 12, further comprising 0.5% (w/w) to 35% (w/w) of an additional component.

14. The composition according to claim 13, further comprising approximately 20% (w/w) of maltodextrin.

15. The composition according to claim 14, further comprising approximately 2% (w/w) of silicon dioxide or tricalcium phosphate.

Description

EXAMPLES

Example 1: Preparation of 10 kg of Crushed Cucumbers Mixed With 30 kg of Purified Water

[0033] The mixture was exposed to a temperature of 70° C. using two extraction steps. The plant matter was separated from the micelle by solid-liquid filtration resulting in a liquid with a solids concentration of 6%. Said micelle underwent a tangential filtration process using a filter cartridge with a nanofiltration membrane (300 Da). The working pressure during the filtration process was between 4-30 bar.

[0034] The result of the filtration was that 80% of the active substance (total amino derivatives, including imino and amino sugars) was recovered in the concentrate phase. The purity of the active substance in the final product was 12% in the permeate phase which meant a purification by 3 times of the initial content.

Example 2: Preparation of 10 kg of Crushed Cucumbers Mixed With 30 kg of Purified Water

[0035] The mixture was exposed to a temperature of 70° C. using two extraction steps. The plant matter was separated from the micelle by solid-liquid filtration resulting in a liquid with a solids concentration of 6%. Said micelle underwent a tangential filtration process using a filter cartridge with a nanofiltration membrane (300 Da). For this process the pH of the micelle was adjusted to a value between 2-5 by the addition of a mineral acid. The working pressure during the filtration process was between 4-30 bar.

[0036] The result of the filtration was that 80% of the active substance (total amino derivatives) was recovered in the concentrate phase. The purity of the active substance in the final product was 17% in the permeate phase which meant a purification by 4 times of the initial content.

[0037] In this case the pH adjustment achieved an unexpected and not initially obvious purification effect in the extract that was filtered through the membrane.

Example 3: Preparation of 10 kg of Crushed Cucumbers Mixed With 30 kg of Purified Water

[0038] The mixture was exposed to a temperature of 50° C. and the plant matter was separated from the micelle by solid-liquid filtration resulting in a liquid with a solids concentration of 6%. Said micelle underwent a tangential filtration process using a filter cartridge with a nanofiltration membrane (300 Da). For this process the pH of the micelle was adjusted to a value between 2-5 by the addition of a mineral acid.

[0039] To aid filtration, 2% of glycerine was added to the mixture and the working pressure during the filtration process was between 4-30 bar.

[0040] The result of the filtration was that 83% of the active substance (total amino derivatives) was recovered in the concentrate phase. The purity of the active substance in the final product was 25% in the permeate phase which meant a purification by 6 times of the initial content.

[0041] In this case the pH adjustment and the addition of glycerine as an additive achieved an unexpected and not initially obvious purification effect in the extract that was filtered through the membrane.

Example 4: Extraction of 5 kg of Magnolia Bark by a Mixture of 60% Alcohol and Water at 55° C. Using Two Extraction Steps

[0042] The plant matter was separated from the micelle by solid-liquid filtration resulting in a liquid with a solids concentration of 4.5% and a flavonolignan content of 2.5%. Said micelle underwent a tangential filtration process using a filter cartridge with a large-pore membrane (1000-10,000 Da) suitable for the passage of organic solvents. For this process the pH of the micelle was adjusted to a value between 5-9 by adding a mineral base. The working pressure during the filtration process was between 4-30 bar.

[0043] The result of the filtration was that in the permeate phase 65% of the initial flavonolignans were recovered which meant a purification by 3 times of the initial content.

Example 5: Extraction of 8 kg of Crushed Turmeric Root With 30 kg of Alcohol Diluted With Water to 85%

[0044] The plant matter was separated from the micelle by solid-liquid filtration resulting in a liquid with a solids concentration of 3.0% and a curcuminoid content of 17%. Said micelle underwent a tangential filtration process using a filter cartridge with a nanofiltration membrane capable of filtering substances dissolved in organic solvents (300 Da). For this process the pH of the micelle was adjusted to a value between 7 and 8.5 by adding an alkaline substance (sodium bicarbonate). The working pressure during the filtration process was between 4-30 bar.

[0045] The result of the filtration was that 76% of the total initial curcuminoids were recovered in the permeate phase. The content in the final product was increased to values of 45% which meant a purification by 3 times of the initial content without the need to use organic solvents other than ethanol.

Example 6: Extraction of 4 kg of Cocoa Powder With Purified Water

[0046] The solid was separated from the liquid by filtration and the resulting liquid was concentrated by evaporation to a dry residue of more than 75%. The concentrate was mixed with ethanol in a ratio of 1 to 10. After stirring, the result was a precipitation of the sugars present in the solution. This solution was then filtered and the precipitate was discarded.

[0047] The liquid was adjusted to a dry residue of 2% with ethanol and passed through nanofiltration membranes capable of working with organic solvents (Organic solvent nanofiltration membranes, OSN). Specifically, a 900 Da membrane was used in diafiltration mode.

[0048] The result of the described process makes it possible to obtain a cocoa extract with a high content of flavan-3-ols and a reduction of the alkaloid content.

[0049] There are conventional methods for performing this reduction of alkaloids in cocoa, which can be contaminating since they use highly hazardous organic solvents such as chlorine products or solvents that can interfere with the aim of preserving the flavan-3-ols in the final product, such as ethyl acetate that would remove the alkaloids and also the flavan-3-ols. The same would happen with water.

[0050] This method is an efficient, environmentally friendly, reproducible way that also preserves the properties of the product.

Example 7: Increase of the flavan-3-ol Content in a Sample of Grape Seed Extract

[0051] To extract the grape seeds, ultrapure water was used as a solvent. The solid was separated from the liquid by filtration and the resulting liquid was concentrated by evaporation to a dry residue of more than 75%. The concentrate was mixed with ethanol in a ratio of 1 to 10. After stirring, the result was a precipitation of the sugars present in the solution. This solution was then filtered and the precipitate was discarded.

[0052] The liquid was adjusted to a dry residue of 2% with ethanol and passed through nanofiltration membranes capable of working with organic solvents (Organic solvent nanofiltration membranes, OSN). Specifically, a 900 Da membrane was used in diafiltration mode.

[0053] The result of this operation was an increase of more than three times in the flavan-3-ol content of the extract.

Example 8: Increase of the flavan-3-ol and Total Polyphenol Content in a Sample of Pine Bark Extract

[0054] To extract pine bark, ultrapure water was used as a solvent. The solid was separated from the liquid by filtration and the resulting liquid was concentrated by evaporation to a dry residue of more than 75%. The concentrate was mixed with ethanol in a ratio of 1 to 10. After stirring, the result was a precipitation of the sugars present in the solution. This solution was then filtered and the precipitate was discarded.

[0055] The liquid was adjusted to a dry residue of 2% with ethanol and passed through nanofiltration membranes capable of working with organic solvents (Organic solvent nanofiltration membranes, OSN). Specifically, a 900 Da membrane was used in diafiltration mode.

[0056] The result of this operation was an increase of more than three times in the flavan-3-ol and total polyphenol content of the extract.

Example 9: Extraction of 10 kg of Spinach Selected so as to Obtain a Product With High Ecdysteroid Content

[0057] The extraction was carried out with water at 65° C. and after that the solid was separated from the liquid by filtration.

[0058] A micelle was obtained with a dry residue of around 3% and a total of 3 kg of dry matter. The ecdysteroid content of the product was 0.19% of the dry matter.

[0059] Said micelle underwent a tangential filtration process using a filter cartridge with a nanofiltration membrane (300 Da). For this process the pH of the micelle was adjusted to a value between 3-5 by the addition of a mineral acid. The working pressure during the filtration process was between 4-30 bar.

[0060] The result of the filtration was that 95% of the initial ecdysteroids were recovered in the concentrate phase compared to the 10% obtained in the permeate phase. This meant an effective purification of around 4 times the ecdysteroid content.

Example 10: Extraction of 10 kg of Lemon Rind With Water at 50° C. for 4 Hours

[0061] The liquid was separated from the undissolved solid by centrifugation. The extract contained the compound of interest, eriocitrin, a flavonoid that is abundant in citrus fruits.

[0062] The micelle with dry residue of 5% underwent a process of purification by tangential cascade filtration. For these, three linked steps were carried out: [0063] 1. The micelle was filtered through a 100,000 Da membrane, with 99% of the eriocitrin passing into the permeate and obtaining 3% of dry residue. [0064] 2. The permeate of the first step was filtered through a 10,000 Da membrane, 99% of the compound of interest passing into the permeate and obtaining 1% of solids. [0065] 3. The permeate of step 2 was filtered through a 1000 Da membrane, 98% of the eriocitrin passing into the permeate and obtaining 0.5% of solids.

[0066] In the three steps as a whole, the liquid extract went from 5% of solids to 0.5% of solids, while maintaining the same amount of eriocitrin. Therefore, it was purified 10 times.

Example 11: Extraction of 10 kg of Lemon Rind With Water at 50° C. for 4 Hours

[0067] The liquid was separated from the undissolved solid by centrifugation. The extract contained the compound of interest, eriocitrin.

[0068] The micelle with dry residue of 5% underwent a process of purification by tangential cascade filtration. For these, three linked steps were carried out: [0069] 1. The micelle was filtered through a 100,000 Da membrane, with 99% of the eriocitrin passing into the permeate and obtaining 3% of dry residue. [0070] 2. The permeate of the first step was adjusted to a basic pH by adding an alkaline substance (sodium hydroxide) and the solution was filtered through a 10,000 Da membrane, 99% of the compound of interest passing into the permeate. The pH caused a higher retention of solids which finally resulted in 0.5% of solids in the permeate. [0071] 3. The permeate of step 2 was filtered through a 1000 Da membrane, 98% of the eriocitrin passing into the permeate and obtaining 0.25% of solids.

[0072] In the three steps as a whole, the liquid extract went from 5% of solids to 0.25% of solids, while maintaining the same amount of eriocitrin. Therefore, it was purified a total of 18 times.

Example 12: Extraction of 10 kg of Lemon Rind With Ultrapure Water at 50° C. for 4 Hours

[0073] Sulphuric ashes are a parameter that measures the content of inorganic cations present in the product. Tap water in many cities tends to contain high conductivity, even above 1000 μS. The use of tap water as extraction water reduces costs in the final product, but entails a series of drawbacks, including the presence of inorganic ions that considerably increase the content of sulphuric ash (Ca.sup.2+, Na.sup.+, K.sup.+), and may be outside the customer's specifications (around a maximum of 1%).

[0074] An extraction equivalent to Example 10 was carried out but with ultrapure water subjected to a double reverse osmosis process from 980 μS to 100 μS in a first phase and to less than 10 μS in a second phase. The extraction was carried out with water with conductivity lower than 10 μS. Once the purification was carried out as described in Example 10, it was dried using a spray-dryer obtaining a solid extract. The sulphuric ashes were analyzed and an extract made with tap water and another with ultrapure water with conductivity lower than 10 μS were compared. The results showed a decrease in the sulphuric ash content of 1.7% (out of the 1% admitted by the majority of customers) to 0.1% of the product made with ultrapure water.

Example 13: Extraction of 10 kg of Lemon Rind With Ultrapure Water at 50° C. for 4 Hours

[0075] Lemon rind was extracted with ultrapure water and the liquid was separated from the undissolved solid by centrifugation. The extract contained the compound of interest, eriocitrin, a flavonoid abundant in citrus.

[0076] The micelle with a dry residue of 5% underwent a process of purification by tangential cascade filtration. For these, three linked steps were carried out: [0077] 1 The micelle was filtered through a 100,000 Da membrane, with 99% of the eriocitrin passing into the permeate and obtaining 3% of dry residue. [0078] 2 The permeate of the first stage was filtered through a 10,000 Da membrane 99% of the compound of interest passing into the permeate and resulted in 1% of solids. [0079] 3 The permeate of step 2 was filtered through a 1,000 Da membrane, 98% of the eriocitrin passing into the permeate and obtaining 0.5% solids.

[0080] The liquid extract with 0.5% of solids is dried by spray dryer, but it is concentrated to 15% of solids to improve drying performance. For this, a concentration stage is carried out using reverse osmosis membranes.

[0081] Reverse osmosis allows the passage of water, but not the rest of the compounds in the extract, eg eriocitrin, or inorganic compounds such as sodium, potassium, or calcium salts, among others. When reverse osmosis concentrates (removes water), the concentration of solutes increases and reaches a point where it is not possible to concentrate more (since the working pressure too high). The use of ultrapure water makes it possible to reach the objective of between 15-20% of dry residue through a final stage of concentration through osmosis. The use of tap water only allows reaching a concentration of between 2.5-7.5% dry residue due to the high content of inorganic salts. The use of ultrapure water would improve the performance in hours of drying, more than 60% with respect to running water.

Example 14: Extraction of 10 kg of Lemon Rind With Ultrapure Water at 50° C. for 4 Hours

[0082] Lemon rind was extracted with ultrapure water and the liquid was separated from the undissolved solid by centrifugation. The extract contained the compound of interest, eriocitrin.

[0083] The micelle with a dry residue of 5% underwent a process of purification by tangential cascade filtration. For these, three linked steps were carried out: [0084] 1 The micelle was filtered through a 100,000 Da membrane, with 99% of the eriocitrin passing into the permeate and obtaining 3% of dry residue. [0085] 2 The permeate of the first stage was filtered through a 10,000 Da membrane 99% of the compound of interest passing into the permeate and resulted in 1% of solids. [0086] 3 The permeate of step 2 was filtered through a 1,000 Da membrane, 98% of the eriocitrin passing into the permeate and obtaining 0.5% solids.

[0087] Another parameter that it is indicative of the purity of the extract obtained is the conductivity measurement, an indicator of the presence of dissolved ions in the water.

[0088] There may be customer requirements that request to assess the content of salts or inorganic compounds, the measurement of conductivity in a 1% solids solution of the dry extract.

[0089] The extract obtained with ultrapure water and dried after its purification, was subjected to the conductivity test, measuring its value in a 1% solution. The conductivity values obtained were between 2.0-2.5 mS compared to 5.0-10.0 mS of an extract made with tap water.

Example 15. Extraction of 15 kg of Persimmon Fresh Fruits With Ultrapure Water

[0090] The fresh fruit persimmon has a dry matter content of 15%, water and high fibre content.

[0091] The extraction was carried out with 45 kg of water at 50° C. and with mechanical stirring for 75 minutes. The high content of fibers and cell walls present in the fruits makes it necessary to use 10 ml of Viscozyme® enzyme complex as an aid to the solid-liquid separation by filtration.

[0092] The use of ultrapure water in the process proved to be extremely useful since it increases the effectiveness of the enzymatic complex, being able to reduce the amount used to half that used with the process carried out with tap water.

[0093] After separation of the extracted solid and the extraction liquid, 49 litres of extract (micelle) were obtained with a dry residue of 2.1%. This extract presents an amount of hydrolyzable tannins of 3.2% determined by HPLC and expressed as epicatechin. Said micelle was subjected to a tangential filtration process using a filtration cartridge with a 300 Da nanofiltration membrane. The result of the filtration was that 90% of the starting hydrolyzable tannins were recovered in the concentrate phase.

[0094] This filtration resulted in a concentration of approximately 13.7 times the tannin content reaching a final purity of 44%.