Process for purifying a compound

Abstract

"PROCESS FOR PURIFICATION OF A COMPOUND". The present invention relates to a process for purifying a compound using an activated carbon treatment. In the process according to the present invention, several filter units containing carbon immobilized in a matrix are operating in series in a counter-current mode. After passing an adequate volume of feed, a filter unit of the first series of filter units is disconnected in a particular position number, and an additional filter unit is connected in another particular position number with this one obtaining a series after the filter units where on the volume change after feeding is continuous. This process overcomes the problem of loss of yield of the purified compound when it occurs during treatment with conventional activated carbon.

Description

PROCESS FOR PURIFYING A COMPOUND The present invention relates to a process for purifying a compound using activated carbon treatment. For many decades, in processes to purify valuable compounds, activated carbon treatments have been applied, whereby bulk activated carbon powder is used to remove impurities such as colored species from the valuable compound. However, the problem when using bulk carbon powder is that activated carbon particles frequently migrate downstream, resulting in carbon contamination in subsequent recovery steps. Also, working with bulk carbon in industrial-scale purification processes does not benefit health and safety. More recently, activated carbon cartridges have been developed that overcome these problems. In these cartridges, activated carbon is immobilized in a filter medium. The use of an activated carbon cartridge is described for the purification of penicillin V (R. Janson and M. Weaver, Manufacturing Chemist, March 2002, pp. 29-30).However, despite the fact that the use of carbon cartridges also eliminates time consumption in recovery processes and leads to improved final product quality, cartridges have not been widely implemented in industrial processes, despite the long-standing need for an improvement over conventional carbon treatment. One of the problems is that the yield of the desired compound is not always favorable enough to operate purification with activated carbon cartridges on a commercially effective industrial scale. The object of the present invention is to overcome this problem of insufficient yield of the desired compound. This problem is solved by the present invention by passing a feed containing the desired compound over a series of filter units containing activated carbon, operating in series and in a counter-current mode. The present invention thus relates to a process for purifying a compound, said process comprising treatment with activated carbon using a filter unit containing activated carbon immobilized in a matrix, the treatment comprising: a) passing an appropriate volume of a feed containing the compound over a first series of n connected filter units operating in series to obtain an effluent, wherein n is at least two, said filter units having been assigned a position number 1 to n in the series and position number 1 being the first supplied with the feed; b) Disconnect a filter unit from the first series of filter units at any position number between 1 and -1 after passing the appropriate volume of feed, and connect a new filter unit at any position that has a higher number than the position number of the disconnected filter unit, resulting in a subsequent series of filter units; c) pass an appropriate volume of subsequent feed containing the compound over the subsequent series of filter units to obtain an effluent; d) optionally combine the effluents obtained in a ec, e Actinomycin, tetracycline, natamycin, clavulanic acid-type β3-lactam compounds, penicillin-G, penicillin-V, cephalosporin C, cefamycin, 6-aminopenicillin (6-APA), 7-aminodeacetoxycephalosporinic acid (7-ADCA), 7-aminocephalosporanic acid (7-ACA), semi-synthetic penicillins such as amoxicillin, cloxacillin, flucloxacillin, methicillin, oxacillin, carbenicillin, ampicillin and semi-synthetic cephalosporins such as sephalexin, cefadrine, cephaloridine, cephalothin, cefaclor, cefacroxil. Carotenoids may include β3-carotene. In a particular embodiment of the present invention, the compound is produced by fermentation using a microorganism. The microorganism may be prokaryotic or eukaryotic, or a cell or cell line of mammalian or plant origin that is capable of producing a compound of interest during fermentation. Preferably, the microorganism is a bacterium, fungus, or yeast. The bacterium may be a strain of E. coli, Streptomyces, Bacillus, or Propionibacterium. The fungus may be a strain of Penicillium, Aspergillus, or Mucor. The yeast may be a strain of Saccharomyces, Kluyveromyces, or Pichia. In a preferred embodiment of the present invention, the compound is obtained by fermentation using a species of Streptomyces as a microorganism. Compounds obtained by fermentation of Streptomyces species are particularly suitable for purification using carbon treatment according to the present invention, provided that a fermentation broth obtained from Streptomyces species, and also the compound produced, contain a colored species and other impurities that may clearly These staining agents can be manifested by a yellow to reddish / brown color. According to the present invention, these staining agents and other impurities are removed very effectively from the compound with a surprisingly high yield of purified compound. Preferred Streptomyces species may be Streptomyces clavuligerus, S. coelicolor, S. griseus, S. vegane, S. aureofaciens. The compounds produced by these strains may be clavulanic acid, streptomycin, cefamycin, chloramphenicol, tetracycline, actinomycin, or β-carotene. Preferably, the compound is clavulanic acid. The fermentation fluid comprising the compound of interest can be separated from the biomass in the fermentation broth by filtration. Optionally, the fermentation fluid comprising the compound can be concentrated and / or the compound can be precipitated or purified using methods known in the art, prior to treatment with activated carbon. The feed that is subjected to treatment with activated carbon according to the present invention contains the compound and includes a solvent. The solvent that is used will typically depend on the compound of interest. It can be water, an alcohol, a ketone, an ester, or a mixture thereof. Preferably, it comprises an alkyl acetate type ester, more preferably ethyl acetate or methyl acetate. The filter units contain activated carbon. immobilized in a matrix. The matrix can be any porous filter medium permeable to the feed containing the compound. Preferably, the matrix comprises a material support material and / or a binding material. js / jer.”.: e .: :" •: : j The support in the matrix can be a synthetic polymer or a polymer of natural origin. The synthetic polymer can include polystyrene, polyacrylamide, polymethyl methacrylate. The polymer of natural origin can include cellulose, polysaccharide, dextran, agarose. Preferably, the polymer support material is in the form of a fiber network to provide sufficient mechanical rigidity. The binder material can be a resin. The matrix can be in the form of a membrane sheet. Preferably, the activated carbon immobilized in a matrix can be in the form of a cartridge. A cartridge is an easily replaceable and self-contained entity containing powdered activated carbon immobilized in a matrix and prepared in the form of a membrane sheet. The membrane sheet can be captured in a permeable plastic support to form a disc. Alternatively, the membrane sheet can be spirally wound. To increase the surface area of ​​the filter, several discs can be stacked on top of each other. Preferably, the discs stacked on top of each other, they have a tube with a core. central for collection and removal of treated feed with Carbon filter unit. Disc configuration. Stacked cartridges can be lenticular. It is also possible to add additional cartridges to an existing filter unit, either by placing these additional cartridges on the same collector shaft (and performing these actions extends the height of the stack) or by accommodating a parallel stack of cartridges in the same filter unit and, optionally, connecting the separate collector shafts to an outlet at the bottom of the unit. Filter housing. Carbon can be obtained from different raw material sources such as peat, lignite, wood, or coconut husk. The choice of carbon source depends on the compound to be isolated and can be determined according to methods known in the art. Any process known in the art, such as chemical or steam treatment, can be used to activate carbon. In the present invention, activated carbon immobilized in a matrix can be placed in a housing to form an independent filter unit. Each filter unit has its own inlet and outlet for the feed containing the compound to be purified. Examples of filter units that are usable in the present invention are the carbon cartridges from Cuno Inc. (Meriden, USA) or Pali Corporation (East Hill, USA). In the invention process, after passing an appropriate volume of feed, a filter unit at any position number between n_n - 1 is disconnected from the series of connected filter units, and a new filter unit is connected at any position that has a position number higher than the position number of the disconnected filter unit (filter unit replacement). The size of the "appropriate volume of feed" (or the timing of filter unit replacement) depends on several parameters and can be determined by optimizing the normal process. For example, the feed volume may depend on the required effluent quality and / or the number of filter units used. Thus, filter unit replacement can occur when a The filter unit used is substantially saturated with Impurities. The moment when it is substantially saturated, being visible by the color of the is not acceptable. which filtering unit is contaminated with impurities can, for example, effluent reaching a value that In a preferred embodiment, the volume of feed passed over the first series of connected filter units n is the same size as the volume of feed subsequently passed over the next series of filter units. In this medium, process logistics are kept as simple and reproducible as possible. In another preferred embodiment, a filter unit can be disconnected at position number 1 and a new filter unit can be connected at position number n + 1. To minimize loss of the compound of interest, the filter unit can be rinsed with solvent before changing the filter unit, preferably using the same solvent in which the compound is dissolved. The solvent rinsing can be preceded and / or followed by purging with a gas, preferably nitrogen. In this way, residual product absorbed into the carbon and / or residual product in the feed volume remaining within the matrix can be recovered. In the process according to the present invention, a feed containing the compound is passed over at least 2 connected filter units operating in series, that is, n is at least 2. Preferably n is 2 to 10. More preferably, n is 2 to 4, most preferably n is 3. Also, several filter units operating in series can additionally, they can be connected in parallel so that 1. Process large feed streams that comprise the compound to be purified. In a particular embodiment of the invention, according to which a series of 2 connected filter units is used, the filter unit in position number one, that is, at the head of the series, is supplied with a feed containing the compound to be purified, and the effluent from this filter unit one is passed over a second filter unit in position number 2. When a suitable volume of feed has passed through the filter units, an in-use change of the filter units is made by disconnecting The filter unit at position number 1 is replaced by a new filter unit at position number 3, resulting in a renumbering of the position numbers, since the original filter unit at position number 2 is now first supplied with power and assigned position number 1, and the filter unit previously at position number 3 is now assigned position number 2. In another embodiment of the invention, when a series of Three connected filter units are used. The filter unit in position one, that is, at the head of the series, is supplied with the feed containing the compound to be purified, and the effluent from this first filter unit is passed over a second filter unit in position two. The effluent from this second filter unit is then passed over a third filter unit in position three. When a suitable volume of feed has passed through the three filter units, a changeover of the filter units is made by disconnecting the unit. filter in position number 1 and connecting a unit A new filter unit at position number 4 results in a subsequent renumbering of the position numbers, since the original filter unit at position number 2 is now first supplied with power and assigned position number 1, and the filter unit previously at position number 3 is now assigned position number 2, and the new filter unit connected at position number 4 is now assigned position number 3. Alternatively, instead of disconnecting the first filter unit, the second (i.e., at position number 2) can be disconnected, resulting in the filter unit at position number 1 remaining there since it is still first supplied with power, and the filter unit at position number 3 is now assigned position number 2, and the new filter unit connected at position number 4 is assigned position number 3 in the second series = The disconnected filter unit in the series is a filter unit containing used activated carbon; that is, feed containing the compound to be purified has been passed over this activated carbon. The new filter unit connected to the series may contain unused activated carbon (i.e., activated carbon not previously used) or it may contain regenerated activated carbon that has been previously used. The new filter unit may be moistened with solvent and subsequently purged with nitrogen before use. Regenerated activated carbon has undergone a regeneration process to recover the absorption capacity of the activated carbon. Regeneration can be carried out by rinsing with a solvent according to the processes. Known in the art. Typical solvents for regeneration may be methanol, ethanol, acetone, or ethyl acetate. Regeneration can occur in situ. When in situ, it means that the filter unit containing the activated carbon to be regenerated is rinsed with a solvent without the need to either physically move the filter unit from its position in the series or physically move the activated carbon from the filter unit. During regeneration, the activated carbon may be subjected to rinsing operations with the solvent present in the previous feed, and / or rinsing with the regeneration solvent, and / or wetting with the solvent present in the subsequent feed. Between rinsing and / or wetting operations, the activated carbon may be purged with gas, preferably nitrogen. In the process according to the present invention, each filter unit can be connected to and disconnected from the series of filter units by physical movement of the unit. Preferably, the filter unit can be connected to and disconnected from the series of filter units without physical movement of the unit. This can be facilitated by a flow distribution system. This flow distribution system can be completely automatic. Preferably, the flow distribution system can comprise multifunctional and multi-port valves, preferably of the shut-off and drain type. The operation of said valves can be controlled by software. More preferably, connection and disconnection of the filter units occur simultaneously. In the present invention, the process can be operated in a batch, semi-continuous or continuous mode. Batch operation means a process whereby a suitable volume of feed is passed over filter units operating in series, and whereby said feed is terminated when a filter unit is disconnected and / or a new filter unit is connected in series. Subsequently, after disconnection and connection (filter unit exchange) has occurred, the feed flow is continuous with a suitable volume of feed following. Continuous operation means a process in which the feed flow is not interrupted when a filter unit is disconnected and a new filter unit is connected, i.e., it is not interrupted when a filter unit is changed. Therefore, a suitable volume of feed and any subsequent suitable volume of feed are continuously passed over the series of connected filter units (n), with filter unit changes occurring at appropriate time intervals. Continuous operation is preferably performed in a situation where it is known what volume of feed can be passed through the series of filter units before a filter unit change occurs. This technology can be obtained through experience or, for example, by online measurement.A prerequisite for continuous mode performance is that the time required for the filter unit changeover operation should be less than the time required to substantially saturate a filter unit with impurities. A continuous process can be terminated, for example, when a change to a new protocol (batch) is desired or for cleaning or other reasons. maintenance . Operating in semi-continuous mode means a process in which the power supply is not interrupted when a filter unit is disconnected and a new filter unit is connected, i.e., it is not interrupted when changing the filter, but the power supply is interrupted to prevent substantial saturation of a filter unit with impurities. This can occur when the time required for the filter unit replacement operation is longer than the time required to substantially saturate a filter unit with impurities. The process according to the present invention can be carried out in a variety of embodiments, all aiming at increased yield of the compound. In a first embodiment, the feed flow rate is 0.05 to 400 l / min, preferably 20 to 100 l / min, more preferably 30 to 40 l / min. The feed flow rate is at least 0.05 l / min. Preferably the flow rate is at least 20 l / min, more preferably the flow rate is at least 30 l / min. The flow rate may have a maximum of 400 l / min. Preferably the flow rate is not above 100 l / min, more preferably the flow rate is not above 40 l / min. In another modality, when the activated carbon immobilized in a matrix is ​​in the form of a membrane sheet with a surface area given in square meters (m²). 2 ), the feed flow rate is 1 to 50 l / m 2 / min, preferably 1.5 to 20 l / m 2 / min, more preferably 1.5 to 10 l / m 2 / min. Preferably, the flow rate is at least 1 l / m 2 / min. More preferably the flow rate is at least 1.5 l / m 2 / min. The flow rate can have a maximum of 50 l / m 2 / min, preferably the flow rate is not above 20 l / min 2 / min, but preferably the flow rate is not above 15 l / min 2 / min. With flow means the feed flow rate per square meter of membrane sheet surface area. In another embodiment, the residence time of the feed containing the compound in a single filter unit is at least 15 seconds and a maximum of 60 seconds. The residence time of the feed containing the compound in a single filter unit is at least 15 seconds, preferably at least 30 seconds, more preferably at least 60 seconds, and even more preferably 2 minutes. The residence time of the compound in a single filter unit is a maximum of 60 minutes, preferably no more than 30 minutes, but preferably no more than 15 minutes. The residence time of the feed containing the compound in a single filter unit can be determined by measuring the difference in time between the feed entering and the feed exiting a single filter unit.When a feed containing the compound is passed over n filter units connected in series, the total residence time of the feed in the series is n times the residence time in a single filter unit. In another mode, the process can be operated at a temperature between -10 and +40°C. It is clear that the temperature is chosen when the feed containing the compound is in the liquid phase both before and after passing through the filter units. The temperature may depend on the type of solvent present in the feed. and the thermostability of the compound. The temperature is at least -10°C, preferably at least -2°C, more preferably 5°C. The temperature may not be more than 40°C, preferably not more than 25°C, more preferably not more than 15°C. After applying the activated carbon treatment according to the present invention, the compound is recovered from the effluent according to processes known to a person skilled in the art. The processes used for recovery will typically depend on the type of compound and / or the intended use. Recovery may include one or a combination of at least two of: stabilizing the compound in the effluent with suitable stabilizing agents, concentrating the effluent, drying the effluent, subjecting the effluent to a granulation process, purifying the compound from the effluent by, for example, crystallization and / or column chromatography. The recovered compound can be further converted into a pharmaceutically acceptable salt or a food-grade product. Example 1 An aqueous broth of clavulanic acid obtained by fermentation of Streptomyces clavuligerus was filtered, extracted, and concentrated to 30 g / L before treatment with activated carbon. 500 mL of concentrated extract were added to a beaker containing 50 g of powdered activated carbon and a magnetic stirrer. After a reaction time of 90 minutes, the activated carbon was separated from the extract using a Bunchner funnel. The percentage of discoloration was determined by measuring the differences in Extinction of the extract before and after treatment with activated carbon in a colorimeter. The percentage of discoloration was 90%. The yield of clavulanic acid after treatment with activated carbon was 86%. Example 2 An aqueous broth obtained from clavulanic acid fermentation was filtered, extracted, and concentrated to 30 g / L before treatment with activated carbon. 500 ml of concentrated extract were passed through a single filter unit containing an activated carbon filter plate (Zetaerbon® R35, 090 mm from CUNO Ltda.) with an approximate effective surface area of ​​0.0057 m². 2 The feed flow rate with concentrated extract over the filter unit was adjusted to 0.03 L / min. The flow rate was 5.0 L / min / m². 2 The percentage of discoloration was 90%. The yield of clavulanic acid after treatment with carbon was 90%. Example 3 A 37.5-liter feed containing concentrated clavulanic acid extract (25 g / L) was passed over 3 connected filter units operating in series, each filter unit containing new activated carbon cartridges (Zetaerbon® C08DB; R35S from CUNO Ltda) approximately 0.29 m long. 2effective surface area. The feed flow rate was 1.0 l / min and the flow rate was 3.5 l / min / m 2 The filter unit at position number 1 was first supplied with feed containing the impure extract. The filter unit at position number 2 was exposed to the effluent from filter unit 1. The filter unit at position number 3 was exposed to the effluent from Filter unit 2. An additional fourth filter unit. It was aligned in the series at position number 4, but it is not in operation since it is not connected to the series of 3 filter units. After passing the 37.5 liters of extract over the three filter units, filter unit number 1 was first supplied with power, then disconnected from the series, and a filter unit previously in position number 4 was connected to the series, resulting in a new assignment of position numbers: the unit previously in position number 4 is now assigned position number 3; the unit previously in position number 3 is now assigned position number 2; the unit previously in position number 2 is now assigned position number 1, to form a second series of 3 connected filter units operating in series, according to which the first and second in the series are both once used and the third filter unit in the series is new.After disconnecting the filter unit and connecting the new one, the feed continued to pass 37.5 liters of non-carbon treated feed containing concentrated clavulanic acid extract over this second series of 3 filter units. In this second filtration, the percentage of discoloration was 94%. The total yield of clavulanic acid was 94. Example 4 The activated carbon treatment from example 3 was repeated with the following difference: the filter units in both the first and second series of 3 connected filter units contained, respectively, a used cartridge twice. times in position number 1, a cartridge used once in Position number 2 and a new cartridge in position number 3. In the second filtration, the percentage of decolorization was 93%. The total yield of clavulanic acid in the collected decolorized extract was 97%. Example 5 The activated carbon treatment was performed according to example 4. After passing the clavulanic acid feed over the second series of filter units, the activated carbon cartridges contained in the filter units were washed by passing 21 l of ethyl acetate over the filter units. The percentage of discoloration was 94%. The total yield of clavulanic acid in the extract The total amount of discolored material collected was 98%.

Claims

CLAIMS 1. A process for purifying a compound, characterized in that it comprises treatment with activated carbon using a filter unit containing activated carbon immobilized in a matrix; the treatment comprises: a) passing an appropriate volume of a feed containing the compound over a first series of n connected filter units operating in series to obtain an effluent, wherein n is at least two, said filter units having been assigned a position number 1 to n in the series and position number 1 being the first supplied with the feed; b) Disconnect a filter unit from the first series of filter units at any position number between 1 and -1 after passing the appropriate volume of feed, and connect a new filter unit at any position that has a higher number than the position number of the disconnected filter unit, resulting in a subsequent series of filter units; c) pass an appropriate volume of feed containing the compound over the series of filter units to obtain an effluent; d) optionally combine the effluents obtained in aec, and e) recover the compound from the effluent.

2. Process, according to claim 1, characterized in that the filter unit is disconnected at position number 1 an - 1 and according to which the new filter unit is connected at position number position n + 1.

3. Process, according to claim 1, characterized in that the filter unit is disconnected at position number 1 and whereby the new filter unit is connected at position number n + 1.

4. A process according to any one of claims 1, 2 or 3, characterized in that the number n of connected filter units operating in series is 2 to 10.

5. Process, according to any one of claims 1, 2, 3 or 4, characterized in that the treatment is operated in batch, semi-continuous or continuous mode.

6. Process, according to any one of claims 1, 2, 3, 4 or 5, characterized in that the feed flow rate is 0.05 to 400 l / min, of Preferably 20 to 100 l / min, more preferably 30 to 40 l / min.

7. Process according to any one of claims 1, 2, 3, 4, 5 or 6, characterized by the fact that where the activated carbon immobilized in a matrix is ​​in the form of a membrane sheet.

8. Process according to claim 7, characterized in that the flow over the membrane sheet is 1 to 50 l / m 2 / min, preferably 1.5 to 20 l / m 2 / min, more preferably 1.5 to 10 l / m 2 / min.

9. Process, according to any one of claims 1, 2, 3, 4, 5, 6, 7 or 8, characterized by the fact that the residence time of the food containing the The filtration time in a single filter unit is at least 15 seconds and a maximum of 60 seconds.

10. Process according to any one of claims 1, 2, 3, 4, 5, 6, 7, 8 or 9, characterized in that the process is operated at a temperature between -10°C and 40°C.

11. A process according to any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, characterized in that at least one unit Disconnected filter media is regenerated in situ by rinsing with an urn solvent.

12. Process according to any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11, characterized in that the compound is an unstable compound.

13. Process according to any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11, characterized in that the compound is an unstable compound. Claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, characterized in that the compound is a secondary metabolite or a protein.

14. Process according to claim 13, characterized in that the secondary metabolite is selected from the group consisting of an antibiotic, a vitamin, a carotenoid or a PUFA.

15. Process, according to any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14, characterized in that the compound is obtained by fermentation using a microorganism.

16. Process according to claim 14, characterized in that the microorganism is a Streptomyces species.

11. Process, according to claim 15, characterized in that the Streptomyces species is selected from the group consisting of S. clavuligerus, S. coelicolor, S. griseus, S. Venezuela, S. jumonjinensis, S. katsurahamanis or S. aureofaciens.

18. Process, according to any one of claims 14, 15, 16 or 17, characterized in that the compound is selected from the group consisting of clavulanic acid, streptomycin, chloramphenicol, tetracycline or O-carotene.

19. A process according to any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18, characterized in that it further comprises the step of converting the compound into a pharmaceutically acceptable salt or quality product. to feed. PROCESS FOR PURIFYING A COMPOUND The present invention relates to a process for purifying a compound using activated carbon treatment. In the process according to the present invention, several filter units containing immobilized carbon in A series of filters are operating in a counter-current mode. After passing a suitable volume of feed, a filter unit from the first series of filter units is disconnected at a particular position number, and an additional filter unit is connected at another particular position number, thus obtaining a series of filter units where the subsequent passage of feed volume is continuous. This process overcomes the problem of yield loss of the purified compound that occurs during treatment with... conventional activated carbon.