WO2001062697A1 - Dihydro and hexahydro isoalpha acids having a high ratio of trans to cis isomers, production thereof, and products containing the same - Google Patents
Dihydro and hexahydro isoalpha acids having a high ratio of trans to cis isomers, production thereof, and products containing the same Download PDFInfo
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- WO2001062697A1 WO2001062697A1 PCT/US2001/005339 US0105339W WO0162697A1 WO 2001062697 A1 WO2001062697 A1 WO 2001062697A1 US 0105339 W US0105339 W US 0105339W WO 0162697 A1 WO0162697 A1 WO 0162697A1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C3/00—Treatment of hops
- C12C3/04—Conserving; Storing; Packing
- C12C3/08—Solvent extracts from hops
- C12C3/085—Extraction of hops with beerwort
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C5/00—Other raw materials for the preparation of beer
- C12C5/02—Additives for beer
Definitions
- IA isoalpha acids
- DHIA dihydro-isoalpha acids
- HHIA hexahydro-isoalpha acids
- IA and THIA do not form insoluble crystalline precipitates upon standing, due to their chemical composition, which includes a keto group on the lower acyl side chain.
- Commercially available all cis isomer DHIA and HHIA have this keto group reduced to an alcohol. They form precipitates over time, which are exceptionally hard to redissolve.
- Their solubility in water at pH 10 is about 1%, and much less at pH 7 to 8.
- the products described herein, containing large amounts of the trans isomers of DHIA and HHIA are remarkably and unexpectedly soluble in water and overcome this limitation, being soluble in water at all concentrations below about 10% to 40%, depending upon the trans isomer content . More detailed description of the prior art:
- IA is sold as a 30% solution of its potassium salt at a pH of about 10 in water.
- DHIA is sold as a 35% solution of its potassium salt in water at a pH of about 10.5 and above, from which large, insoluble crystals of DHIA will precipitate over time.
- THIA is used as a 5% or 10% solution of its potassium salt at a pH of about 9.5 to 10.5 in water; and HHIA is not sold as an aqueous solution per se because of its limited solubility.
- the hop acids and particularly 30% IA and 35% DHIA, as potassium salts at pH 10 or above in water, act as co-solvents for themselves.
- the co-solvent effect is demonstrated by the known tendency to precipitate and separate at lower concentrations, as discussed below under the Westermann prior art.
- all forms of hop acids can be solubilized in propylene glycol, as described in Todd (U.S. P. 3,486,906), and are available in this form, which also adds the advantage of increasing their dispersibility in soft water at pH 10 and above.
- Propylene glycol and ethanol solutions are the only forms of HHIA available, and their utility is impaired by the requirement of a solvent .
- the high trans products overcome the need to use propylene glycol or ethanol as a solvent .
- soft water must be used as the diluting agent for all potassium salt solutions of the iso acids, since calcium and magnesium in the water will form chelates with hop acids and cause a haze and agglomerates and gummy precipitates.
- Below pH about 9 to 10 in deionized water the dilute solutions of the prior art DHIA and HHIA do not form a clear solution upon mixing but rather form gummy precipitates upon standing. The high trans products do not.
- hop acids post-fermentation One common method of adding the hop acids post-fermentation is to dilute them to a 1% or less concentration in soft water to which KOH has been added to bring the pH to 10 or above (Held, Master Brewers of the Americas
- suspensions were made from commercial all cis products ( ⁇ l2, 1 23-24) made by the prior art procedures described herein. They are suspensions. They had the advantage over the prior art commercial solutions of DHIA in that they did not require heating to about 80-90° C to redissolve precipitates before use. Indeed, one of the major advantages was the ability to redissolve the micro-particles by heating to about 60°C .
- the redissolved solution was in turn diluted to 1% in soft water at a pH of 10 prior to injection in the beer.
- the micro particles could be added directly to pH 10 soft water preheated to about 50 °C, wherein they would dissolve and form a clear 1% solution within five minutes.
- the 1% solution forms a haze upon standing (page 23, line 5), while the high trans product does not. Furthermore, his product, like other prior art products, is not soluble at a 1 % concentration in neutral soft water, whereas the products described herein are completely soluble. And furthermore, his product still required heating, albeit less vigorous than 80-90° C.
- the novel high trans product can preferably be used at ambient temperature, including brewhouse cellar temperatures of 10°C or less. The commercialization of his product was abandoned because of its limitations in practical brewing use, and particularly the need to heat it and the lack of clarity upon dilution.
- DHIA is made from alpha acids by isomerization and reduction using sodium borohydride, first described by Koch (U.S. . 3,044,879).
- Koch U.S. . 3,044,879
- a superior process based on Koch was described in Westermann (U.S. P. 3,798,332), which used an extract made by his earlier invention (U.S. P. 3,558,326).
- Goldstein U.S. P. 4,324,810) describes a method of making DHIA without the use of organic solvents.
- manufacturers optionally separate the alpha acids from the remainder of the extract prior to isomerization and reduction, as described in Goldstein U.S. P. 4,767,640. These investigators produced the essentially all cis forms of the acids.
- Todd U.S. P. 4,002,683 describes an improved method for separation of alpha acids and subsequent isomerization to IA, which is the preferred method of separating alpha acids from an extract.
- a less desirable procedure for the separation of alpha acids from an extract and conversion to IA is given in Klingel, (U.S. . 3,364,265), who also describes solid salts of IA.
- Mitchell U.S. P. 3,949,092 describes a superior process.
- the method of separating and purifying the alpha acids is not critical to the disclosed process.
- the purity of the reduced product is a critical element.
- the ratio of trans to cis isomers is very critical, and new to the art.
- THIA Procedures for making THIA from alpha acids are described in Stegink (U.S. P. 5,296,637), and Hay, (U.S. P. 5,013,571).
- THIA is made from beta acids after the procedure of Worden, (U.S. . 3,923,897).
- HHIA is made by borohydride reduction after the method of Todd (U.S. P. 4,666,731, Example 10), who employs less than half the molar equivalents of Worden, (U.S. P. 3,552,975) to achieve reduction in a highly alkaline medium.
- Hay also describes the catalytic reduction of cis DHIA to make cis HHIA.
- HHIA like DHIA, must be substantially free of impurities if it is to form the novel product of the present invention. None of these investigators have suggested this aspect of the present invention.
- Guzinski (U.S. P. 5,200,227) describes mixtures of the prior art concentrated aqueous products which, due to co-solvent effects, do not readily crystallize. These had the advantage of physical stability over the single acid products, but imposed limitations on the ratios of different acids which the brewer could add to a beer. Occasionally, it was found that large crystals would form from these mixtures after prolonged storage, but not to the extent formed in the single-acid forms of commerce. Because of the limitations on the ratios of the different hop acids, they have limited utility. These products formed two phase, gummy solutions upon dilution in water, just as do prior art 35% DHIA solutions. The novel forms of DHIA and HHIA described herein overcome these limitations, since they are non-crystallizing and do not form gummy particulates .
- Bavisotto U.S. P. 3,615,660 describes the use of e ulsifiers to stabilize DHIA extracts and make them suitable for adding to wort or beer.
- the instant products overcome the need for the use of emulsifying agents which end up in the beer, and the precipitation of the DHIA extract as the emulsion breaks upon addition to the beer.
- the aroma is derived from the essential oil contained in the hop cone.
- Aroma control is compatible with this invention by addition of hop essential oil to the kettle (preferably in the saponified extract described in Guzinski, USP 5,750,179). This invention also allows the addition of hop essential oil to the DHIA and HHIA solutions, wherein it is sufficiently soluble to enable the brewer to add controlled amounts of essential oil to the finished brew.
- the objects of this disclosure are to provide DHIA and HHIA having a high trans to cis isomer ratio and, as a consequence, to provide: 1. A non-precipitating solution of DHIA and / or HHIA. 2. Non-precipitating mixtures of DHIA and / or HHIA solutions with added IA and THIA. 3. Hop acid solutions which do not form a haze or particulates upon direct injection into finished beer. 4. The analytical criteria which will provide quality assurance for the products, and which differentiates them from all prior art products. 5. The operational variables which may be adjusted by the manufacturer when making the novel products.
- hop acids do not form precipitates which must be heated to redissolve, or which must be filtered from the beer. They are soluble in soft water and their dilute solutions will not form hazes in the brewing cellar injection tank. Because the purity of the hop acid must be high to make them clear, they do not contribute an off-flavor "hang" to the beer, but rather possess only the desired fleeting bitterness without after-bitter, especially on the palate. They can be directly injected into finished beer without forming haze or visible particulates, contrary to prior art products.
- the preparation of the product critically differs from the prior art in that the reduction is performed in an aqueous medium with sodium borohydride (potassium borohydride is less preferred) at a pH below about 12, and preferably in the range of about 10 to 11, and at temperatures, times and concentrations which do not convert trans isomers to cis isomers.
- aqueous medium with sodium borohydride (potassium borohydride is less preferred) at a pH below about 12, and preferably in the range of about 10 to 11, and at temperatures, times and concentrations which do not convert trans isomers to cis isomers.
- Prior art products are made using a more highly alkaline aqueous medium (pH 13.5), since it is well known that borohydrides decompose readily if the water in which they are dissolved is not highly alkaline.
- the presently- disclosed and critical procedure allows some borohydride to decompose due to the lower pH, while the remainder acts as a reducing agent. Buffers may be used
- the effect of the lower pH on the DHIA or HHIA is to allow trans isomers to form without being changed to the cis isomer. It increases the critical ratio of trans to cis isomers. Unless the trans isomer HPLC area count is at least 10% of the cis isomer area count, and preferably greater than about 20 to 30%, the product will not form a clear liquid aqueous solution at all concentrations from 1% to 20% and more. This is critical to the invention. Prior art products have a ratio of trans to cis isomers of less than about 3% to 5% and, in most, trans isomers are undetectable. None of them will form clear solutions at concentration ranges of 10-20% in water, even at elevated pHs . The novel solubility properties of high trans isomer ratio containing DHIA and HHIA are disclosed for the first time in this specification. The preferred method also involves the reduction of
- Goldstein in U.S. P. 4,767,640 separates the alpha acids from the extract, at a marginally higher pH than the critical pH of Todd, prior to isomerization/reduction without the use of solvents. He obtains an improved product, devoid of non-isohumulone light unstable products (NILUPS) found in the prior art products.
- NILUPS non-isohumulone light unstable products
- Westermann claimed complete light stability, it is clear that the detection of instability had progressed by the time of Goldstein's invention, and he was able to improve the light stability of Westermann's products .
- His product is claimed suitable for post-fermentation addition to beer, but not specifically for pre- or post- final filtration. This may be because his product forms amorphous agglomerates and crystals on standing.
- innocuous solvents such as hydrocarbons C-10 and below are useful in assisting the separations and purifications of the high trans products. They are not essential but rather optional and will assist in the removal of the unwanted impurities, some of which are visible as post hop acid peaks in the HPLC. Others, such as "waxes", may be undetectable in the HPLC assay. These must be substantially absent for the claimed DHIA and HHIA to remain clear in aqueous solutions when added to soft water. Chicoye et al, in U.S. P. 4,759,941, describe a method for making DHIA by treating hop pellets with borohydride.
- Example 9 shows the differences in performance of the products in beer.
- HHIA hexahydro-isoalpha acids
- DHIA dihy- droisoalpha acids
- HHIA hexahydro-isoalpha acids having a ratio of trans to cis isomers greater than 10%; such a mixture comprising hexahydro-isocoalpha acids, hexahydro-iso-n-alpha acids, and hexahydro-isoadalpha acids; such a mixture wherein the ratio is greater than 20%; such a mixture wherein the ratio is greater than 40%; and such a mixture wherein the ratio is greater than 70%.
- HHIA hexahydro-isoalpha acids
- such a mixture in the form of an aqueous solution of potassium salts of the HHIA which solution forms a single phase liquid at a 20% concentration by weight of the potassium salts at a pH less than 9.5; such a mixture wherein the solution forms a single phase liquid at a 10% concentration by weight of the potassium salts of the HHIA at a pH less than 8.5; such a mixture in the form of an aqueous solution of the potassium salts of the HHIA at a pH of 7 to 10.5 which is a single-phase solution when at a concentration of 5% by weight; such a mixture in the form of an aqueous solution of the potassium salts of the HHIA at a pH of 7 to 9.5 which is a single-phase solution when at a concentration of 10% by weight; and such a solution which, when diluted to a 1% concentration by weight in distilled water, forms a clear solution which does not form a haze upon standing for six hours .
- IA isoalpha acids
- THIA tetrahydroisoalpha acids
- DHIA dihydro-isoalpha acids having a ratio of trans to cis isomers greater than 10%; such a mixture comprising dihydro-isocoalpha acids, dihydro- iso-n-alpha acids, and dihydro-isoadalpha acids; such a mixture wherein the ratio is greater than 20%; such a mixture wherein- the ratio is greater than 30%.
- such a mixture in the form of an aqueous solution of potassium salts of the DHIA which solution forms a single phase liquid at a 20% concentration by weight of the potassium salts at a pH less than 9.5; such a mixture wherein the solution forms a single phase liquid at a 10% concentration by weight of the potassium salts of the DHIA at a pH less than 8.5; such a mixture in the form of an aqueous solution of the potassium salts of the DHIA at a pH of 7 to 10.5 which is a single-phase solution when at a concentration of 5% by weight; such a mixture in the form of an aqueous solution of the potassium salts of the DHIA at a pH of 7 to 9.5 which is a single-phase solution when at a concentration of 10% by weight; and such a solution which, when diluted to a 1% concentration by weight in distilled water, forms a clear solution which does not form a haze upon standing for six hours ; Also, such a mixture which contains less than 5% by
- such a mixture of DHIA or HHIA which is in the form of a single-phase aqueous solution of its potassium salts at a pH above about 7.5 when at a concentration of 20% by weight.
- the process of reducing (a) isoalpha acids (IA) to produce dihydroisoalpha acids (DHIA) or (b) tetrahydroisoalpha acids (THIA) to produce hexahydroiso- alpha acids (HHIA) the DHIA or the HHIA product having a trans to cis isomer ratio greater than 10%, the reduction being carried out in an aqueous medium at a pH of about 8.5 to about 12.4 using a borohydride; such a process wherein IA are reduced to DHIA having a trans to cis isomer ratio greater than 10% using less than about 0.81 molar equivalents of a borohydride and a pH up to about 11.8; such a process wherein THIA are reduced
- HHIA phase is concentrated at a pH below about 10.5 and greater than 6 by evaporation of water, to give a concentrated aqueous phase containing between about 5% and about 40% DHIA or HHIA; such a process wherein the pH is between 6.5 and 8.5 and the DHIA or HHIA concentration is less than about 25-s; such a process wherein the borohydride is selected from the group consisting of sodium borohydride and potassium borohydride; and such a process wherein the DHIA having a trans to cis isomer ratio greater than 10% is subsequently converted to HHIA having a trans to cis isomer ratio greater than 10% by catalytic hydrogenation.
- trans and cis isomers of the hop analogues exist.
- Critical to this invention is the heretofore unknown effect of a high trans: cis isomer ratio on the solubility of the DHIA and HHIA.
- this ratio is expressed as the % of the HPLC area counts of the trans divided by the area counts of the cis isomers. In prior art products it is well below 5% and usually almost zero.
- UV spectra UV Spectra
- Hops-6 American Society of Brewing Chemists spectro procedure
- This entails diluting the test sample in alkaline methanol and running a scan, and using a formula to calculate % alpha acids. This procedure is included in the prior art references .
- Absorption at 254 nm is a maximum for iso acids, and the strength of the sample is calculated on this basis using the extinction coefficient (El%/lcm) .
- the sample is dissolved in alkaline methanol, the absorbance at
- the extinction coefficient will vary for the various hop acids due to differences in molecular weights, analogue composition, and the standards historically used to determine them. For the purposes of this specification, the following numbers are used:
- Hop extracts are diluted to a concentration of about
- the detector is set to measure the entire UV absorbance spectrum between 230-400 nm with a resolution of 1.2 nm, filter response of 1, and sampling rate of 1 point/sec.
- HPLC plots are reported in "maxplot" mode, which reports the maximum absorbance value between 230- 400 nm at each point in the chromatogram.
- Data is analyzed by Millennium ® 32 software (version 3.05.01, Waters and Associates) .
- the % impurities eluting after the hop acid is determined using the % area count at peak maximum. This is because many of the impurities do not have significant absorbance at 254, but peak in the range of 270 nm and above. An extinction coefficient is not needed for this calculation, as it only measures the total area under the peaks at the absorption maximum.
- the subject hop acids are identified in the traces, as well as the peaks eluting after them, and the instrument calculates the area counts .
- the relative area counts are independent of concentration of the solution injected into the HPLC.
- Yields are based on an average molecular weight of the mixture of analogues .
- Infra-red (IR) spectra are useful for demonstrating the different chemical composition of the pure hop acid and the haze forming substances which do not absorb uv light. For the purposes of this specification, they are defined as "waxes.” These are isolated and the spectra described in Example 11. GENERAL DESCRIPTION OF THE INVENTION
- Examples 1 thru 4 show variations on the preferred process for making a high trans , highly soluble DHIA and HHIA which, in turn, do not form hazes upon dilution to 1% in distilled water..
- hop acids The purity of the hop acids must be exceptionally high if solutions of the high trans product are to remain clear. Substances eluting after the hop acid in the HPLC procedure must be less than 6%, preferably less than 4%, and most preferably less than 1% to 2%. Likewise, "waxes'
- trans to cis isomer ratio of above about 10%, especially above about 20% to 30%, results in the greatly increased solubility of the cis isomers, which, as mentioned in Example 6, are shown to be about 1.5% maximum for equally pure cis DHIA and 0.75% for cis HHIA.
- the trans isomers increase the solubility of the cis isomers from about 1% in water to 10% and more. For example, a 20% solution of HHIA containing 4% trans isomers and 16% cis isomers does not form crystalline precipitates. A 16% solution of cis isomers does. As mentioned above, the solubility of the cis isomers alone is about 1%.
- the effect of the trans isomers upon the solubility of the cis isomers' solubility is contrary to expectation, since the higher the solute content, the lower should be the solubility of related compounds .
- This effect cannot be a simple result of the analogue mixture, since the analogues are the same for the cis and trans forms.
- the claimed products contain the approximate mixture of analogues found in the parent hop. Neither of these critical elements-the high trans isomer ratio combined with all of the parent hop analogues-- have grounding in the prior art .
- Edible anti freeze substances such as ethanol, propylene glycol, and glycerine may be added to the inventive products if they are to be exposed to below freezing temperatures.
- buffering agents other than potassium phosphate may be used. These include sodium and other phosphates, as well as borates and citrates. Details concerning the required process parameters are discussed in the Examples.
- the claimed products When the claimed products are dried, they form amorphous solids which can readily be rehydrated to form aqueous solutions with the properties of the original aqueous solutions.
- Dehydration can be performed by techniques known to the art, such as spray drying or by evaporation of water under vacuum or even at atmospheric pressure.
- the claimed product is differentiated from prior art products by its high trans isomer content, the trans isomers being at least about 10% of the cis isomers (a trans to cis ratio of 10%) , and preferably 20%, and most preferably above 30%. It is further differentiated by the absence of substances which elute after the DHIA or HHIA by HPLC analysis, such substances consisting of artifacts and by-products of the reduction reaction. These substances interfere with the clarity of the aqueous solutions of the products.
- the products are further differentiated from the prior art in that substances which are soluble in hydrocarbon solvents and not detected by the HPLC procedure are essentially absent .
- the products form stable single phase aqueous solutions at pHs substantially below the 10.5 to 11 minimums of the prior art, for example between about 7 and 9.5, and are not dependent upon a 35% hop acid concentration, as shown by Westermann, to make a pourable liquid product.
- the solutions are stable at concentrations in the range of about 5% to 40%. While the instant products are preferably maintained at a pH below about 9.5, they are also stable at the pHs of the prior art .
- the products form clear, non-hazing solutions in distilled water at concentrations of 1% to about 5% and more.
- Prior art products require raising the pH of the water to above 10 to effect dispersion of a 1% solution, and even then haze forms upon standing. They form gums and precipitates when added to distilled water. This simple test is one means of determining if the product meets the analytical requirements described above and in the Claims.
- Reaction temperatures below about 85° C are feasible, the reaction taking longer at lower temperatures.
- the preferred range is about 40° to 75° C.
- the reduction should be terminated before a significant amount of trans isomer is converted to the cis form. This occurs more rapidly at high pHs and temperatures .
- the analytical procedures described herein provide a guide to termination times.
- non-uv absorbing substances undetectable by the HPLC procedure, which must be critically less than about 3%, preferably less than 2%, and most preferably less than 1% of the weight of the hop acids.
- waxes Removal of these unwanted and unidentified substances, called “waxes” in this specification, and not absorbing uv light, is preferably achieved by separating them from aqueous solutions at a pH below 10.5, and preferably below about 8.5 to 9.5, and even as low as 7.5.
- concentration of the hop acids in the aqueous phase during "wax" removal is less than about 20%, and preferably less than 15%. Because of the insolubility of the all cis prior art forms at these pHs, separations were done at elevated temperatures (Goldstein) or less than about half of the DHIA was captured into the "clean" phase.
- yields were poor and sufficient impurities were present to cause haze and precipitation when diluted in distilled water. It is speculated that the presence of the trans forms assists in the separations, and therefore is critical to the "clean-up" procedure.
- the herein disclosed art gives yields in excess of 75% and up to 85% to 90%. Separation of unwanted substances is preferably effected using a hydrocarbon solvent, especially of C-6 to C-10, but other water immiscible solvents such as ether or methylene chloride may be used.
- the solid separations are conducted at a pH below about 9.5.
- FIG. 1 is a depiction of the structural formulas of cis and trans IA and DHIA.
- FIG. 2 is a depiction of the structural formulas of cis and trans THIA and HHIA.
- FIG. 3 is a trace of a HPL Chromatogram of typical prior art all-cis DHIA plotting absorbance units against time.
- FIG. 4 is a trace of a HPL Chromatogram of trans DHIA plotting absorbance units against time.
- FIG. 5 is a trace of a HPL Chromatogram of high trans DHIA plotting absorbance units against time.
- FIG. 6 is an overlay of the chromatograms of FIGS. 3 and 5.
- FIG. 7 is a trace of a HPL Chromatogram of a cis HHIA commercial product plotting absorbance units against time.
- FIG. 8 is a trace of a HPL Chromatogram of trans HHIA plotting absorbance units against time.
- FIG. 9 is a trace of a HPL Chromatogram of high trans HHIA plotting absorbance units against time.
- FIG. 10 is an overlay of the chromatograms of FIGS. 7 and 9. DETAILED DESCRIPTION OF THE INVENTION
- Example 1 Preparation of a liquid DHIA by reduction without the use of solvent in the reducing medium.
- the organic phase containing the DHIA, was then partitioned against 100 ml of warmed distilled water brought to a pH of 8.4 with potassium hydroxide.
- a pH of between about 7.6 to 8.8 is optimal for this separation, although higher and lower pHs may be used.
- concentrations of 20%, preferably 15%, and most preferably about 10 % or less in the aqueous phase are suitable for the partition.
- the aqueous phase was reextracted with hexane to remove "waxes" and other insoluble material.
- the aqueous layer was then desolventized and concentrated to a 19% solution under vacuum.
- the 19% aqueous solution was soluble with complete clarity at all concentrations in distilled water, and the trans isomers HPLC area count was 33% of the cis isomer area count (trans: cis ratio equals 33%). The yield was 74%.
- the 19% solution remained clear, single phase, and without precipitates, unlike any prior art product.
- Charcoal and silica gel and other adsorbents are useful alternatives for water immiscible solvents for removing the substances which detract from the clarity of the soluble DHIA solutions.
- Lower alkanols such as methanol, ethanol, and isopropanol may be present in the reaction medium. They will speed the reaction, but will also decrease the ratio of trans isomers .
- the prior art products consisting of 35% DHIA aqueous liquids wherein the hop acids are substantially free of trans isomers, as in the prior art examples below, are insoluble in water, and form a haze following addition to pH 10 water.
- Example 2 Preparation of DHIA from IA with the use of solvent .
- 500g of a 25% (measured by total absorbance at 254 nm) aqueous solution of crude IA was added to 250 ml of a 4% solution of NaBH 4 (0.75 molar equivalents) .
- the pH was 11.4.
- 125ml of limonene, a C-10 hydrocarbon was added, and the mixture heated with agitation for 3 hours at 70-75° C.
- the mixture was cooled, and the organic phase separated from the aqueous DHIA phase. The organic phase was discarded.
- aqueous DHIA phase was concentrated under vacuum to 20% DHIA, during which process all residual limonene was removed.
- the DHIA had a trans: cis ratio of 24% as measured by HPLC area counts.
- the absorbance, as measured at peak maxima, of the post-DHIA eluting substances was about 2.0 to 2.5% of the total DHIA area count.
- the post-DHIA eluting substances were about 9.9% in the crude reaction mixture prior to partitioning. This shows the effectiveness of the partition in removing the unwanted, haze forming impurities) .
- the yield was about 70% of the crude IA.
- the 20% aqueous DHIA solution remained clear and without crystallization for more than three months. It formed clear solutions at all concentrations when diluted in distilled water.
- hexane be used during the boron removal procedure .
- the HHIA was recovered from the hexane solution by partition into water with dilute KOH to a pH between about 6.5 and 8.5, preferably and in this example about 7.5, to form an aqueous solution of HHIH.
- the concentration was about 10%. Its optional range is about 5% to 15% and less preferably 20% HHIA.
- the organic phase was separated from the aqueous HHIA phase and discarded.
- the aqueous phase and about 10% by volume of hexane were heated to reflux with agitation. Temperature is preferably elevated, and can be the reflux temperature of hexane or other water immiscible solvent, the reflux temperature being easily controlled.
- the aqueous phase is then concentrated to any desired % HHIA by removal of water under vacuum, which also assures removal of any residual solvent.
- the resulting aqueous phase was
- HHIA 13.4% HHIA by uv.
- the peaks eluting after HHIA were 1.77% of the HHIA peaks by area count at peak maximum, and HHIA was 92.1% of the area count at peak maximums .
- the balance was material eluting prior to HHIA, which does not interfere with the solubility. It had a trans: cis ratio of 98%. It did not form precipitates on standing for three months .
- HHIA with the above solubility characteristics and a trans: cis ratio of above 10% may also be made by catalytic hydrogenation of DHIA with a tran:cis ratio of above 10%. However, this is not a preferred procedure.
- Example 4 Reduction of THIA to HHIA in a buffered system.
- the aqueous phase was discarded.
- the hexane phase was reextracted at pH about 3 and then with water, pH 4, to remove residual boron.
- the hexane-organic phase was then extracted into 100 ml of water by adjusting the pH to about 8 with 10% KOH.
- the aqueous phase was reextracted twice with hexane, which was discarded.
- the aqueous phase was then rotovapped to provide a 22.5% HHIA solution by uv. It assayed 85% HHIA by HPLC, with 0.44% post-HHIA peaks, 14.6% pre-HHIA peaks, and the HHIA had a trans: cis ratio of 75%. It remained as a clear, light tan solution without the formation of crystalline materials for more than three months .
- potassium phosphate is a preferred buffer
- other buffers such as mixtures of potassium and boron salts may be used.
- the advantage of using a buffer is the maintenance of pH within a narrow range, thus avoiding the effect of variations in pH as the unreduced form is converted to the reduced form, which has a different pKa.
- Example 5 Compatible single phase liquid, non-precipitating mixtures of soluble DHIA and HHIA with IA and THIA .
- IA and THIA do not form crystalline precipitates, perhaps due to their molecular structure. Since the soluble forms of DHIA and HHIA have been unknown to the art, this example is designed to demonstrate the limits of compatibilities of the mixtures. Guzinski U.S. P. 5,200,227 shows the limits of compatible mixtures of IA, DHIA, THIA, and HHIA which do not form precipitates of DHIA or HHIA on standing. The objective of his invention was to overcome the tendency of liquid solutions of DHIA and HHIA to form insoluble precipitates of the hop acids on standing. His formulations have found limited use in the art, since upon standing precipitates may form, particularly with cycles of heating and cooling such as occur during transportation.
- Pure recrystallized cis DHIA has an upper solubility limit of about 1 to 1.5% in distilled water, and pure recrystallized cis HHIA's upper limit is about 0.75% even at pH 10.
- the presently inventive products form clear solutions in distilled water at the concentrations noted above, as well as below 1%. If a high trans DHIA or HHIA at a concentration above about 20 to 25%, for example 35% to 40%, is desired to minimize shipping costs, any precipitates can be dissolved easily by dilution and agitation.
- the high trans DHIA has very different physical properties than essentially all cis prior art DHIA.
- the prior art DHIA precipitates very slowly at a 35% concentration, and separates into two phases at less than about 30%.
- the high trans DHIA remains without precipitates at concentrations below about 25%. The reason for this difference in physical and solubility characteristics is unknown.
- the impurities have an important effect upon the clarity of the 1% solution.
- the optimal product has a trans ratio above 10%, and preferably above 20%, and especially above 30%,
- the DHIA and HHIA impurities eluting after the hop acid by HPLC, as measured by area count at peak maximums, are desirably less than about 8%, and especially less than about 5%, and most preferably less than about 3%, and most desirably less than 1%.
- bitterness profiles of the high trans products were shown to be the same as those of the existing commercial types by two techniques, both of which involve dilution of the test material in water at a concentration of 15 ppm.
- the first is a triangle test, in which individuals are asked to select the odd sample among three samples presented to them. The results are then subjected to statistical analysis to determine if the samples are different within a 95% confidence limit. This test showed that there was no difference in bitterness between the commercial products and the inventive samples.
- the second test is more sophisticated, and tells whether or not the hop acid has the same fading characteristics in the mouth. It is called a time-intensity analysis, and involves the subject tasting a sample in water, and recording the bitterness impression at five second intervals. This provides an analysis of the maximum bitterness perceived, as well as the manner in which the bitterness disappears in the mouth.
- the inventive high trans to cis ratio sample had the same perceived intensity as one commercial DHIA sample, which was more bitter than a second commercial DHIA sample, but otherwise had the same type of fading curves. All had similarly shaped time intensity curves. High trans ratio HHIA samples also had the same time intensity curves as all cis HHIA.
- Example 9 Comparative clarity and solubilities upon injection into beer.
- the products of the above examples were directly injected into a commercial beer at brewing cellar temperature of about 4-8°C . Because the Guzinski PCT suspensions were too thick to inject directly, all formulations were diluted to 1% concentration prior to injection.
- the prior art products, including the microparticles of Guzinski did not form clear solutions upon standing, even though the pH of the solutions was above 10, and was substantially above that of the high trans to cis isomer ratio DHIA and HHIA solutions (pH about 7 to 8) .
- the bottles were shaken, held overnight, and haze measured using a clear glass bottle. The increase in FTU haze units is shown below in Table 9-1. Table 9-1. Behavior ,of products upon injection into beer.
- Example 10 Comparison of HPLC differences between commercial all cis hop products and the novel high trans isomer DHIA and HHIA products.
- Figure 3 is an HPLC trace of a typical prior art all cis- DHIA.
- Fig. 4 is a trace of trans DHIA, which contains a very small amount of cis.
- Fig. 5 is a trace of the new, highly soluble, high trans DHIA.
- Fig. 6 is an overlay of Fig. 3 and Fig. 5, in which the solid line is the cis product, and the dotted line is the new high trans soluble product.
- Figures 7, 8, 9 and 10 show the comparable traces for HHIA.
- Fig. 7 is the trace of the prior art HHIA
- Fig. 8 of all trans HHIA Fig. 9 of the new high trans HHIA
- Fig. 10 an overlay of Fig. 7 and Fig. 9.
- trans isomers convert to cis isomers upon refluxing at pH 12.5 in water. Their identity was further confirmed by using this procedure to produce the cis forms.
- the new high trans DHIA product of Fig. 5 has a large peak, eluting at about 12 % minutes, which is present as a shoulder in the all cis product of Fig. 3, and essentially absent in the all trans DHIA of Fig. 4.
- This peak is an n-analogue cis - stereoisomer which becomes the 16 minute n - analogue cis isomer upon heating at a pH above that used to make the high-trans product. It will be noted that this peak is very evident in the overlay of Fig. 6. Its presence does not affect the identification of the trans peaks.
- Peaks appearing after the last trans isomer are those which are considered post- DHIA and HHIA impurities. Since there are many of them, all with very low area counts in these purified products, they look and are insignificant. In unpurified product, such as commercial DHIA, or an unpurified product, they would be clearly visible and represent about 8% to 15% and more of the area count .
- Example 11 Separation and analysis of "waxes", and haze contributed thereby.
- the aqueous phase was concentrated to 12% HHIA, which removed residual heptane. It formed a clear 1% solution in distilled water, without the formation of hazes upon standing. This demonstrates that the haze forming "waxes" had been removed, and the HHIA will not form haze upon injection into beer as a 1% aqueous solution, which indeed is the case.
- the "waxes" by themselves caused a haze measured at 30 FTU when 2 ppm were injected as a 1% alcoholic solution into beer.
- the conventional dewaxing is done by separating insoluble materials from solutions of HHIA at concentrations above about 15% to 20%. To remove the last traces of haze forming substances, it is preferable to dilute the HHIA to less than 15%, preferably 12-14%, and most preferably below about 10% concentration. Unless the solutions are dilute, more than three extractions of the aqueous phase with the organic solvent are required. It is considered, based on this experiment, that the
- HHIA acts as a cosolvent for the "waxes," which is why HHIA concentrations of less than about 15% facilitate their removal This is why they have not been successfully removed from the prior art all cis isomer commercial products.
- a pH 7.8 DHIA solution with a % trans: cis ratio of 42 was extracted twice with hexane at a concentration of about 15% and the aqueous phase separated from the organic phases. The organics were discarded and the aqueous phase concentrated and desolventized to give a clear 20.7% solution. 227.6g of this clear solution was diluted to a 10% concentration in water. The solution remained clear. Upon dilution in water to 1%, it developed a very slight haze. It was then warmed to 60°C.
- the separated aqueous phase was reduced in volume under vacuum to remove residual hexane, and diluted to 10% and 1% in distilled water. No haze formed upon standing, showing that the haze forming waxes had been removed. Injection of the 1% DHIA solution into beer did not cause an increase in FTU at 10 ppm, whereas the waxes caused an increase of between 10 and 20 FTU at 2 ppm when injected as a 1% alcoholic solution. This again demonstrates that when the purified product does not throw a haze in dilute aqueous solutions, it will not cause a measurable haze when injected into beer. After the aqueous solution was concentrated to 12% DHIA, it remained as a clear solution and did not form precipitates.
- the IR spectra were obtained on a Perkin Elmer 1710 FTIR spectrometer using thin films of the "waxes" cast on KBr windows .
- the spectra showed the presence of OH groups at 3200-3600 and 1000-1300 cm “1 ; aliphatic CH groups at 2800-3000 cm “1 , and carbonyl CO groups at 1600- 1750 cm “1 .
- the OH and carbonyl CO stretching bands are greatly enhanced in pure samples of DHIA and HHIA, while the "waxes" have significantly less absorbance between 2800 and 3000 cm “1 . This is interpreted to mean than the "waxes" contain substantial amounts of aliphatic hydrocarbon moieties in the form of fatty alcohols, which are absent in the pure hop acids. These aliphatic hydrocarbon moieties explain the haze forming potential of the "waxes” .
- Westermann builds on the work of Koch, who showed DHIA in alcoholic solutions which, when added to wort, improved the light stability of beer. Westermann' s objective was to provide a DHIA fraction suitable for post-fermentation addition, and of improved purity.
- the procedure of -326 was followed on a bench scale by combining 50g of a supercritical C02 hop extract (46.5% alpha by uv, 23.25 g) , 175 ml of water, 75 ml of hexane, and 13 g of commercial SWS (12% NaBH 4 in 40% NaOH, 0.63 molar equivalents).
- the pH was 13.5.
- the mixture was heated at 60°C (140° F) for three hours, cooled, and acidified to pH 2 with dilute H 2 S0. It was agitated warm for one hour, and the phases separated.
- the hexane-DHIA phase was washed again with acidic water, and then with water to remove residual boron and acid..
- the hexane-DHIA phase was made to 140 ml with hexane, and 20 ml aliquots were withdrawn for the -188 experiments 11-15, which provided his best yields and most "pure" product. Each aliquot is calculated to contain about 3.32g of DHIA.
- hexane solution was added 20 ml of distilled water, and the pH adjusted upward with a small amount of dilute KOH to the target pH.
- the aqueous phase was separated from the hexane phase, which contained resins, beta acids , and other unwanted substances .
- the aqueous phase was extracted with methylene chloride (10ml) at a pH of 2-3. At this pH the hop substances are extracted into the solvent . The methylene chloride was separated, removed under vacuum, the solids weighed and then assayed by uv and HPLC.
- the weight amounts extracted into the water increase with the pH, as did those of Westermann. This is because the solubility of the hop acids in water increases with pH.
- the uv purities are below his , being measured by the total absorbance at 254nm.
- the HPLC purities are below the uv purities. This is because the HPLC separates the impurities, such as humulinic acid and alpha and beta acids, from the DHIA. Only the DHIA content is used in the HPLC purity calculation. Since humulinic acid has a lower molecular weight than DHIA, and is extracted into water at a lower pH, a higher concentration of that acid will result in a higher uv "purity", such as claimed by him.
- Peaks appearing after the last trans isomer are those which are considered post- DHIA and HHIA impurities. Since there are many of them, all with very low area counts in these purified products, they look and are insignificant. In unpurified product, such as commercial DHIA, or a slightly impure product, they would be clearly visible and represent about 8% to 15% and more of the area count .
- trans isomers convert to cis isomers upon refluxing at pH 12.5 in water. Their identity was further confirmed by using this procedure.
- hop extract 46.5% alpha acids by uv, and 45% KOH (46.6 g) were agitated and warmed to about 45 deg for about 5 minutes, placed in a separatory funnel, and the lower somewhat alkaline aqueous phase containing the alpha acids was separated. It contained 19.4% alpha by uv, as compared to Goldstein's 16.8%.
- reaction mixture was agitated at 175°F (79° C) for 2 hours .
- the boron was removed from the aqueous phase by acidification with 50% H 2 S0 4 with agitation, separation of the lower aqueous layer from the upper acidic DHIA oily phase, and by rewashing of the oily phase at a pH of about 2 with water, and then with water alone.
- the separation and agitation was facilitated by heating to reduce the viscosity of the oil.
- 6.7 g of the acidic DHIA was agitated with water and the pH adjusted with dilute KOH to make a 40% DHIA solution by uv at pH 7.0. It was agitated at 50-65°C. for one hour.
- the aqueous phase was allowed to separate from the NILUPS oil phase as the mixture cooled, and the phases separated in a funnel.
- the DHIA phase assayed 37% DHIA by uv.
- the HPLC showed that this consisted of 83.4% DHIA, and 7.1% post-DHIA peaks by area count at peak maxima. It was not "pure" DHIA. It did not contain trans DHIA.
- the warm liquid DHIA phase separated into two phases, with amorphous semi-crystalline solids appearing overnight . It was reheated to dissolve the crystals prior to evaluation in water, in which it formed a hazy dispersion at pH 10 and concentration of 1%. It formed a two phase solution on cooling before crystals appeared. It formed a hazy dispersion in distilled water.
- Example 15 Comparative with Goldstein -640 Example 1 plus 3.
- Example 1 The procedure in his Example 1 was followed, which differs from his Example 2 in that about 0.74 instead of 0.5 mole equivalents of NaBH 4 is used for reduction.
- the acidic DHIA-NILUP oil phase was separated from residual boron using acidic water. It was titrated with dilute KOH to a pH of 7.0-7.2. , as in his example.
- the aqueous DHIA phase was separated, after cooling overnight, from the NILUP oil phase.
- the DHIA aqueous phase assayed 41.5% DHIA by uv, and the DHIA was 80.2% of the HPLC 254nm area count.
- Example 3 The acid form as recovered in his Example 3 assayed 78.2% DHIA, and 8.2% post-DHIA by area count at peak maxima. He does not provided HPLC assays for these final products. Trans DHIA was not detected. Yields were consistent with those reported by Goldstein. The warm liquid DHIA phase formed amorphous solids overnight. It was reheated to dissolve the crystals prior to evaluation in beer in Example 9.
- Example 16 Discussion of effect of varying conditions upon the reduction .
- the conditions which produce a high trans product are similar to those of the prior art, in that reaction times, temperatures, and pHs, and equivalents of borohydride are found.
- the reason that the high trans product has not been produced in the prior art is simply that the right combination of conditions has not been used. Indeed, the thrust of the art has been to produce the 35% all cis DHIA first described by Westermann.
- the prior art conditions under which the reductions were performed were not critical, in that the principal objective was to make sure that essentially all IA was reduced and the light stability improved. This meant that high pHs and temperatures, as well as long reaction times, gave acceptable yields without residual IA, since cis DHIA is stable at high pHs .
- HHIA humulinic acid type and other degradation products were measured as DHIA by uv analysis, yields as measured by uv at 254nm were considered acceptable.
- Commercial production of HHIA essentially follows the procedure of Todd (U.S. P. 4,666,731), which is similar to Westermann, in which SWS (12% sodium borohydride in 40% NaOH) is used, and the pH during the reduction is above 13. The products are all cis isomers.
- trans DHIA or trans HHIA The high pH reduction will not make either trans DHIA or trans HHIA. This is because they "epimerize,” which means that they isomerize to the cis form under the time, temperature, and/or pH conditions of the prior art. Furthermore, the proper balance between time, temperature, pH, and borohydride equivalents must achieve essentially complete reduction of the IA if the DHIA is to be light stable, and yet not epimerize the trans isomer to the cis one, or form undesirable and haze forming over-reduced products. The following rules can be applied to guide one skilled in the art: (1) the lower the pH, the longer the reaction time. (2) the higher the pH, the easier to eliminate residual IA.
- Marginally acceptable DHIA in acceptable yield can be made at a pH of up to about 11.8. Above that pH, formation of the cis isomer predominates, especially above 50° C. Increasing the mole equivalents of borohydride beyond 0.75 moles increases by-product formation to above 15%. By reducing the pH to 11, and the time to two hours, little epimerization occurs and byproduct formation is about the same as at pH 11.7. By reducing the pH to 10.5 , and the temperature to 50° C, epimerization becomes negligible but the time required to achieve about the same residual IA is 5 to 7 hours.
- Table 16-1 compares the prior art ranges with feasible and optimal conditions for reduction of IA and THIA. It will be noticed that the combination of preferred conditions do not coincide with the prior art . For HHIA, which epimerizes more slowly than DHIA, it is noticed that the pH can safely be raised above the pH for DHIA
Abstract
Description
Claims
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DE60128331T DE60128331D1 (en) | 2000-02-25 | 2001-02-20 | DIHYDRO AND HEXAHYDRO ISOALPHIC ACIDS HAVING A HIGH TRANSACTION TO CIS ISOMERS, PROCESS FOR THEIR PRODUCTION AND THESE PRODUCTS CONTAINING THEREOF |
EP01910969A EP1263704B1 (en) | 2000-02-25 | 2001-02-20 | Dihydro and hexahydro isoalpha acids having a high ratio of trans to cis isomers, production thereof, and products containing the same |
CA002400177A CA2400177C (en) | 2000-02-25 | 2001-02-20 | Dihydro and hexahydro isoalpha acids having a high ratio of trans to cis isomers, production thereof, and products containing the same |
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US09/512,944 US6583322B1 (en) | 2000-02-25 | 2000-02-25 | Dihydro and hexahydro isoalpha acids having a high ratio of trans to cis isomers, production thereof, and products containing the same |
US09/512,944 | 2000-02-25 |
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US8206753B2 (en) * | 2001-06-20 | 2012-06-26 | Metaproteomics, Llc | Anti-inflammatory botanical products for the treatment of metabolic syndrome and diabetes |
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US20040115290A1 (en) * | 2001-06-20 | 2004-06-17 | Tripp Matthew L. | Modulation of inflammation by hops fractions and derivatives |
US7270835B2 (en) * | 2001-06-20 | 2007-09-18 | Metaproteomics, Llc | Compositions that treat or inhibit pathological conditions associated with inflammatory response |
US7901713B2 (en) * | 2001-06-20 | 2011-03-08 | Metaproteomics, Llc | Inhibition of COX-2 and/or 5-LOX activity by fractions isolated or derived from hops |
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US20040219240A1 (en) * | 2001-06-20 | 2004-11-04 | Babish John G. | Anti-inflammatory pharmaceutical compositions for reducing inflammation and the treatment or prevention of gastric toxicity |
US8168234B2 (en) | 2001-06-20 | 2012-05-01 | Metaproteomics, Llc | Compositions that treat or inhibit pathological conditions associated with inflammatory response |
US7718198B2 (en) * | 2001-06-20 | 2010-05-18 | Metaproteomics, Llc | Treatment modalities for autoimmune diseases |
US7815944B2 (en) * | 2001-06-20 | 2010-10-19 | Metaproteomics, Llc | Anti-inflammatory pharmaceutical compositions for reducing inflammation and the treatment of prevention of gastric toxicity |
US20090263522A1 (en) * | 2001-10-26 | 2009-10-22 | Babish John G | Curcuminoid Compositions Exhibiting Synergistic Inhibition Of The Expression And/Or Activity Of Cyclooxygenase-2 |
US7279185B2 (en) * | 2001-10-26 | 2007-10-09 | Metaproteonics, Llc | Curcuminoid compositions exhibiting synergistic inhibition of the expression and/or activity of cyclooxygenase-2 |
US8158160B2 (en) | 2001-11-13 | 2012-04-17 | Eric Hauser Kuhrts | Anti-inflammatory cyclooxygenase inhibitors |
NZ539642A (en) * | 2002-10-21 | 2007-01-26 | Metaproteomics Llc | Compositions containing extracts from hops in combination with a second component for treating inflammatory response |
GB0306568D0 (en) * | 2003-03-21 | 2003-04-30 | Unilever Plc | Compositions of natural products and use thereof |
KR20060105429A (en) * | 2003-05-22 | 2006-10-11 | 메타프로테오믹스, 엘엘씨 | Anti-inflammatory pharmaceutical compositions for reducing inflammation and the treatment or prevention of gastric toxicity |
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US7914831B2 (en) | 2004-02-27 | 2011-03-29 | Metaproteomics, Llc | Synergistic anti-inflammatory pharmaceutical compositions and related methods using curcuminoids or methylxanthines |
US20050192356A1 (en) * | 2004-02-27 | 2005-09-01 | Babish John G. | Synergistic anti-inflammatory pharmaceutical compositions and methods of use |
EP1817941A4 (en) * | 2004-11-13 | 2009-11-11 | Metaproteomics Llc | Compositions exhibiting inhibition of cyclooxygenase-2 |
WO2006065131A1 (en) * | 2004-11-22 | 2006-06-22 | Universiteit Leiden, Faculteit Van Wiskunde En Natuurwetenschappen | Method of improving the stability of hop extracts and hop flavoured beverages |
AU2006280074A1 (en) * | 2005-08-09 | 2007-02-22 | Metaproteomics, Llc | Protein kinase modulation by hops and acacia products |
JP2009518439A (en) * | 2005-12-09 | 2009-05-07 | メタプロテオミクス,エルエルシー | Protein kinase regulation by hops and acacia products |
EP2046353A4 (en) * | 2006-06-20 | 2010-01-27 | Metaproteomics Llc | Tetrahydro-isoalpha acid based protein kinase modulation cancer treatment |
US20080051466A1 (en) * | 2006-06-20 | 2008-02-28 | Metaproteomics, Llc | Isoalpha acid based protein kinase modulation cancer treatment |
MX2009010049A (en) * | 2007-03-19 | 2010-03-04 | Metaproteomics Llc | Methods and compositions for promoting bone and joint health. |
WO2008140842A1 (en) * | 2007-05-11 | 2008-11-20 | Metaproteomics, Llc | Methods and compositions for heavy metal detoxification |
AU2008335156A1 (en) * | 2007-12-10 | 2009-06-18 | Metaproteomics, Llc | Substituted 1,3-cyclopentadione multi-target protein kinase modulators of cancer, angiogenesis and the inflammatory pathways associated therewith |
JP2011516492A (en) * | 2008-04-02 | 2011-05-26 | メタプロテオミクス, エルエルシー | Attenuation of endothelial inflammation and endothelium-monocyte interaction by substituted 1,3-cyclopentadione |
US9796955B2 (en) * | 2010-05-24 | 2017-10-24 | S.S. Steiner, Inc. | Natural and stable solutions of alpha-acids and their use for the improvement of foam quality of beer |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3552975A (en) * | 1965-03-01 | 1971-01-05 | Kalamazoo Spice Extract Co | Hop flavors for malt beverages and the like |
US5767319A (en) * | 1996-10-30 | 1998-06-16 | Miller Brewing Company | Preparation of tetrahydroiso-α-acids by the hydrogenation of the metal salts of iso-α-acids |
Family Cites Families (27)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3044879A (en) | 1959-02-11 | 1962-07-17 | Miller Brewing | Anactinic malt product and hop extract therefor |
US3364265A (en) | 1962-08-15 | 1968-01-16 | Steiner Inc S S | Hop constituents and method of making same |
US3486906A (en) | 1965-03-01 | 1969-12-30 | Kalamazoo Spice Extract Co | Water-dispersible hop flavors for malt beverages and the like |
US3949092A (en) | 1965-11-17 | 1976-04-06 | Bush Boake Allen Limited | Hop extract containing potassium isohumulate |
GB1161787A (en) | 1965-11-17 | 1969-08-20 | Bush Boake Allen Ltd | Manufacture of Hop Extracts. |
US3558326A (en) | 1967-09-21 | 1971-01-26 | Miller Brewing | Process for isomerizing and purifying hop extracts |
US3615660A (en) | 1969-06-11 | 1971-10-26 | Pfizer | Hop extract emulsion and preparation and use thereof |
CA945088A (en) | 1970-09-17 | 1974-04-09 | Alexander W. White | Isomerised hop extract |
US4002683A (en) | 1971-05-03 | 1977-01-11 | Kalsec, Inc. | Process for isomerizing alpha acids to iso-alpha acids |
GB1423129A (en) | 1971-08-04 | 1976-01-28 | Bush Boake Allen Ltd | Isomerised hop extracts |
US3965188A (en) | 1972-01-10 | 1976-06-22 | Miller Brewing Company | Hop extract process and product |
US3798332A (en) | 1972-01-10 | 1974-03-19 | Miller Brewing | Preparation of hot extracts and use in beer making operation |
US3923897A (en) | 1973-04-02 | 1975-12-02 | Kalamazoo Spice Extract Co | Production of hoplike beverage bittering materials |
AU476799B2 (en) | 1974-02-04 | 1976-08-05 | Carlton And United Breweries Limited | Preparation of a water soluble isomerised hop extract |
ZA792350B (en) * | 1978-05-26 | 1980-05-28 | Brewing Patents Ltd | Production of iso-alpha acids |
CH649778A5 (en) * | 1978-06-20 | 1985-06-14 | Mueller Adam | METHOD FOR ISOMERIZING ALPHA ACIDS IN A HOP EXTRACT. |
US4324810A (en) | 1980-05-29 | 1982-04-13 | Miller Brewing Company | Hop extracts and method of preparation |
US4590296A (en) * | 1984-01-25 | 1986-05-20 | Miller Brewing Company | Process for separation of beta-acids from extract containing alpha-acids and beta-acids |
US4666731A (en) | 1984-02-28 | 1987-05-19 | Kalamazoo Holdings, Inc. | Separation of the constituents of CO2 hop extracts |
US4759941A (en) | 1985-04-30 | 1988-07-26 | Miller Brewing Company | Anactinic hopping materials and method of preparation |
US4767640A (en) | 1985-10-29 | 1988-08-30 | Miller Brewing Company | Light stable hop extracts and method of preparation |
US5013571A (en) | 1990-01-31 | 1991-05-07 | Pfizer Inc. | Methods for making tetrahydroisoalpha and hexahydroisoalpha acids |
US5200227A (en) | 1992-05-11 | 1993-04-06 | Kalamazoo Holdings, Inc. | Stable aqueous solutions of tetrahydro and hexahydro iso-alpha acids |
US5296637A (en) | 1992-12-31 | 1994-03-22 | Kalamazoo Holdings, Inc. | Production of odor-free tetrahydroisohumulates from alpha acids via their tetrahydrohumulates and subsequent isomerization |
US5750179A (en) | 1994-09-13 | 1998-05-12 | Kalamazoo Holdings, Inc. | Process for producing a water-soluble lipidic hop extract |
US5600012A (en) * | 1995-04-06 | 1997-02-04 | John I. Haas, Inc. | Process for producing tetrahydroisoalpha acids |
WO1997033971A1 (en) | 1996-03-15 | 1997-09-18 | Kalamazoo Holdings, Inc. | Solid hop acid salt compositions |
-
2000
- 2000-02-25 US US09/512,944 patent/US6583322B1/en not_active Expired - Lifetime
-
2001
- 2001-02-20 EP EP01910969A patent/EP1263704B1/en not_active Expired - Lifetime
- 2001-02-20 CA CA002400177A patent/CA2400177C/en not_active Expired - Lifetime
- 2001-02-20 DE DE60128331T patent/DE60128331D1/en not_active Expired - Lifetime
- 2001-02-20 WO PCT/US2001/005339 patent/WO2001062697A1/en active IP Right Grant
- 2001-02-20 AT AT01910969T patent/ATE361908T1/en not_active IP Right Cessation
- 2001-07-23 US US09/911,063 patent/US6521796B2/en not_active Expired - Lifetime
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3552975A (en) * | 1965-03-01 | 1971-01-05 | Kalamazoo Spice Extract Co | Hop flavors for malt beverages and the like |
US5767319A (en) * | 1996-10-30 | 1998-06-16 | Miller Brewing Company | Preparation of tetrahydroiso-α-acids by the hydrogenation of the metal salts of iso-α-acids |
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EP1263704B1 (en) | 2007-05-09 |
US6521796B2 (en) | 2003-02-18 |
US20020007084A1 (en) | 2002-01-17 |
CA2400177A1 (en) | 2001-08-30 |
CA2400177C (en) | 2007-09-18 |
EP1263704A4 (en) | 2004-08-25 |
DE60128331D1 (en) | 2007-06-21 |
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US6583322B1 (en) | 2003-06-24 |
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