Method for removing N-nitroso-vancomycin in vancomycin hydrochloride

Abstract

The invention provides a method for removing N-nitroso-vancomycin in vancomycin hydrochloride, and relates to the technical field of medicine quality control. The removal method comprises the following steps: carrying out first mixing on a first filler and a first solvent to obtain a first solid-liquid mixture; performing first filtration on the first solid-liquid mixture, and collecting a first solid phase; carrying out second mixing on vancomycin hydrochloride to be treated and a second solvent to obtain a first solution; adjusting the pH value of the first solution to enable the first solution to be acidic to obtain a second solution; performing third mixing on the first solid phase and the second solution to obtain a second solid-liquid mixture; performing secondary filtration on the second solid-liquid mixture, and collecting a liquid phase, namely vancomycin hydrochloride product liquid; wherein the first filler is a reverse-phase filler. By utilizing the removal method, the N-nitroso-vancomycin is removed to the maximum extent, so that the concentration of the N-nitroso-vancomycin in the vancomycin hydrochloride is stably and effectively reduced.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical quality control technology, specifically relating to a method for removing N-nitroso-vancomycin from vancomycin hydrochloride. Background Technology

[0002] Vancomycin hydrochloride is a glycopeptide antibiotic with antibacterial activity, primarily effective against Gram-positive bacteria. It is mainly purified from *Streptomyces orientalis*. The mechanism of action of vancomycin hydrochloride is to inhibit bacterial cell wall synthesis, thereby affecting bacterial cell membrane function and interfering with protein and RNA synthesis to achieve a bactericidal effect. Due to its unique mechanism of action, vancomycin hydrochloride is easily degraded in vivo and has few side effects, thus it is widely used clinically and is generally regarded as "the last line of defense against drug-resistant strains." Vancomycin hydrochloride is currently the first-line drug for treating serious infectious diseases such as pneumonia, sepsis, and surgical trauma caused by drug-resistant Staphylococcus aureus. The structural formula of vancomycin hydrochloride is shown in Formula 1.

[0003]

[0004] Vancomycin hydrochloride contains a secondary amine structure in its chemical structure. In the presence of nitrosamines (such as nitrites), it undergoes a nitrosation reaction to form N-nitroso-vancomycin, as shown in Formula 2. N-nitroso-vancomycin is a nitrosamine impurity with a structure similar to vancomycin hydrochloride. Such impurities exhibit high mutagenicity and carcinogenicity even at extremely low exposure levels. In recent years, domestic and international drug regulatory agencies have required safety risk assessments of nitrosamine impurities in related products and have mandated strict control over the N-nitroso-vancomycin content in vancomycin hydrochloride to ensure the safe use of marketed drugs.

[0005]

[0006] Currently, there are three methods for removing N-nitrosovancomycin from vancomycin hydrochloride. The first method is to directly remove its precursors, that is, to replace raw materials such as nitrites with other raw materials to avoid the reaction between vancomycin hydrochloride and nitrosamines from the source. However, this method is difficult to avoid the accumulation of trace amounts of N-nitrosovancomycin in intermediates or finished products generated during the reaction process. The second method is to block the formation pathway of N-nitrosovancomycin by adding vitamin C, vitamin E, etc. However, this method is difficult to effectively reduce the concentration of N-nitrosovancomycin in the product. The third method is to use activated carbon to adsorb N-nitrosovancomycin, but this method has problems such as unstable removal effect and is difficult to achieve large-scale application.

[0007] Therefore, how to provide a method for removing N-nitrosovancomycin from vancomycin hydrochloride with significant removal effect is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0008] The main objective of this invention is to provide a method for removing N-nitrosovancomycin from vancomycin hydrochloride, thereby solving the problem of unstable removal effect in the prior art and thus stably and effectively reducing the concentration of N-nitrosovancomycin in vancomycin hydrochloride.

[0009] To achieve the above objectives, according to one aspect of the present invention, a method for removing N-nitrosovancomycin from vancomycin hydrochloride is provided, the method comprising the following steps:

[0010] The first filler and the first solvent are mixed to obtain a first solid-liquid mixture;

[0011] The first solid-liquid mixture is subjected to a first filtration to collect the first solid phase;

[0012] The vancomycin hydrochloride to be treated is mixed with the second solvent to obtain the first solution;

[0013] Adjust the pH of the first solution to make it acidic to obtain the second solution;

[0014] The first solid phase is mixed with the second solution in a third mixing process to obtain a second solid-liquid mixture.

[0015] The second solid-liquid mixture is filtered a second time, and the liquid phase is collected, which is the vancomycin hydrochloride product solution.

[0016] The first packing material is a reverse-phase packing material.

[0017] This invention involves pretreating the first packing material and acidifying the vancomycin hydrochloride solution. The pretreated packing material is then mixed with the acidified vancomycin hydrochloride solution, allowing N-nitrosovancomycin to be adsorbed. Solid-liquid separation removes the packing material adsorbed with N-nitrosovancomycin, reducing the impurity content in the vancomycin hydrochloride product solution and thus stably and effectively lowering the concentration of N-nitrosovancomycin in vancomycin hydrochloride. Furthermore, this invention utilizes a reverse-phase packing material as the adsorption host. The reverse-phase packing material has a relatively compact structure, good stability, and facilitates subsequent processing and packing material recycling, enabling its reuse.

[0018] Furthermore, the particle size of the first filler is 2–200 μm, and the pore size is [missing information].

[0019] By limiting the particle size of the first packing material to the above range, not only can the specific surface area of ​​the first packing material be increased, that is, the contact area between the vancomycin hydrochloride solution to be treated and the first packing material be increased, thereby improving the adsorption efficiency and adsorption effect, but the loading capacity of the first packing material can also be guaranteed, ensuring that there is enough space to support N-nitrosovancomycin.

[0020] Furthermore, the first packing material is selected from one or more of LSi-200, UniPS10-300, UniPS20-300, UniPS 30-300, UniSil C18, UniSil C8, and UniSil C4. These packing materials have stable physicochemical properties, are insoluble in acids, alkalis, and organic solvents, and can effectively adsorb N-nitrosovancomycin, thereby achieving a stable and effective reduction in the concentration of N-nitrosovancomycin in vancomycin hydrochloride.

[0021] Further, the first solvent is a mixture of an organic solvent and water, wherein the organic solvent is selected from at least one of methanol, ethanol, and acetone; the second solvent is water; preferably, the volume concentration of the organic solvent in the first solvent is 40-80%.

[0022] Furthermore, the temperature of the first mixing is 0–35°C, the time is 0.5–6 h, and the rotation speed is 200–2000 rpm / min. By limiting the parameters such as temperature, time, and rotation speed of the first mixing, it is ensured that the first packing material and the first solvent are in full contact, removing impurities from the surface of the first packing material and avoiding affecting its subsequent adsorption effect on N-nitrosovancomycin.

[0023] Furthermore, the concentration of vancomycin hydrochloride in the first solution is 50,000–150,000 μg / mL.

[0024] Furthermore, the pH value of the second solution is 1.0 to 5.0.

[0025] Furthermore, the mass ratio of the first packing material to the vancomycin hydrochloride to be treated is (1:8) to (5:1). By limiting the mass ratio of the first packing material to the vancomycin hydrochloride to be treated, sufficient first packing material is ensured for adsorbing N-nitroso-vancomycin, while simultaneously ensuring removal efficiency.

[0026] Furthermore, the temperature of the third mixing is 0–35℃, the time is ≥0.5h, and the rotation speed is 200–2000 rpm / min. By limiting the parameters such as temperature, time, and rotation speed of the third mixing, it is ensured that the first solid phase and the second solution are in full contact, so that the first solid phase can adsorb N-nitrosovancomycin in the second solution to the maximum extent, thereby achieving the purpose of improving the removal effect.

[0027] Furthermore, the pH of the first solution is adjusted using an acid adjuster, wherein the acid adjuster is selected from at least one of formic acid, acetic acid, trifluoroacetic acid, hydrochloric acid, phosphoric acid, and nitric acid; preferably, a 0.2-1.0 mol / L acetic acid solution or hydrochloric acid solution is used as the acid adjuster.

[0028] The implementation of this invention has at least the following beneficial effects:

[0029] The present invention provides a method for removing N-nitrosovancomycin from vancomycin hydrochloride. This method utilizes a first filler, after treatment with a first solvent, to adsorb the acidified vancomycin hydrochloride solution, thereby maximizing the removal of N-nitrosovancomycin and stably and effectively reducing the concentration of N-nitrosovancomycin in vancomycin hydrochloride. Specifically, the content of N-nitrosovancomycin in vancomycin hydrochloride can be reduced to below 0.20 ppm. Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0031] As described in the background section of this invention, the existing technology suffers from unstable removal efficiency. To address this issue, this invention provides a method for removing N-nitrosovancomycin from vancomycin hydrochloride. The method includes the following steps: mixing a first packing material with a first solvent to obtain a first solid-liquid mixture; filtering the first solid-liquid mixture to collect the first solid phase; mixing the vancomycin hydrochloride to be treated with a second solvent to obtain a first solution; adjusting the pH of the first solution to make it acidic to obtain a second solution; mixing the first solid phase with the second solution to obtain a second solid-liquid mixture; filtering the second solid-liquid mixture to collect the liquid phase, which is the vancomycin hydrochloride product solution; wherein the first packing material is a reverse-phase packing material.

[0032] In this process, the first packing material and the first solvent are mixed. During the first mixing process, the first solvent is used to pretreat the first packing material to remove impurities from its surface and prevent them from affecting subsequent processing. After the first solid-liquid mixture is filtered, a first solid phase and a first liquid phase are obtained. The first solid phase is the pretreated first packing material, and the first liquid phase is the pretreated first solvent.

[0033] The vancomycin hydrochloride to be treated is mixed with a second solvent to obtain a first solution; alternatively, the vancomycin hydrochloride to be treated is dissolved in a second solvent to form a vancomycin hydrochloride solution to be treated, and the pH value is adjusted to make it acidic to obtain a second solution.

[0034] The first solid phase and the second solution are mixed in a third process to obtain a second solid-liquid mixture. During this third mixing process, N-nitrosovancomycin is adsorbed onto the surface and / or interior of the first solid phase, and the N-nitrosovancomycin in the second solution is removed. The second solid-liquid mixture is then subjected to a second filtration to obtain a second solid phase and a second liquid phase. The second solid phase is the first solid phase containing adsorbed N-nitrosovancomycin, and the first liquid phase is the vancomycin hydrochloride product solution.

[0035] This invention involves pretreating the first packing material and acidifying the vancomycin hydrochloride solution. The pretreated packing material is then mixed with the acidified vancomycin hydrochloride solution, allowing N-nitrosovancomycin to be adsorbed. Solid-liquid separation removes the packing material adsorbed with N-nitrosovancomycin, reducing the impurity content in the vancomycin hydrochloride product solution and thus stably and effectively lowering the concentration of N-nitrosovancomycin in vancomycin hydrochloride. Furthermore, this invention utilizes a reverse-phase packing material as the adsorption host. The reverse-phase packing material has a relatively compact structure, good stability, and facilitates subsequent processing and packing material recycling, enabling its reuse.

[0036] In some embodiments, the particle size of the first filler is 2–200 μm, and the pore size is [missing information]. Specifically, the first packing material serves as the adsorption host. By limiting the particle size of the first packing material to the above-mentioned range, not only can the specific surface area of ​​the first packing material be increased, that is, the contact area between the vancomycin hydrochloride solution to be treated and the first packing material be increased, thereby improving the adsorption efficiency and adsorption effect, but the loading capacity of the first packing material can also be guaranteed, ensuring that there is enough space to support N-nitrosovancomycin.

[0037] This invention does not limit the specific type of the first packing material, as long as the first packing material meets the above requirements. In some embodiments, the first packing material is selected from at least one of macroporous adsorption resin, reverse polymer packing material, and reverse silica gel packing material. These packing materials have stable physicochemical properties, are insoluble in acids, alkalis, and organic solvents, and can effectively adsorb N-nitrosovancomycin, thereby achieving a stable and effective reduction in the concentration of N-nitrosovancomycin in vancomycin hydrochloride.

[0038] The macroporous adsorption resin can be a non-polar macroporous adsorption resin, mainly composed of styrene and propionate monomers, which are cross-linked and polymerized under the action of a cross-linking agent, such as LSi-200. The reverse polymer filler can be selected from UniPS 10-300, UniPS20-300, UniPS 30-300, etc., and the reverse silica gel filler can be selected from UniSil C18, UniSil C8, UniSil C4, etc. Specifically, the first filler can be one or more of LSi-200, UniPS10-300, UniPS20-300, UniPS 30-300, UniSil C18, UniSil C8, and UniSil C4.

[0039] In some embodiments, the first solvent is a mixture of an organic solvent and water, wherein the organic solvent is selected from at least one of methanol, ethanol, and acetone; the second solvent is water; preferably, the volume concentration of the organic solvent in the first solvent is 40-80%, for example, a range of 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, or any combination thereof.

[0040] In some embodiments, the temperature of the first mixing is 0 to 35°C, for example, a range of 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C or any two of these; the time is 0.5 to 6 hours, for example, a range of 0.5 hours, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours or any two of these; and the rotational speed is 200 to 2000 rpm / min, for example, 200 rpm / min, 400 rpm / min, 600 rpm / min, 800 rpm / min, 1000 rpm / min, 1200 rpm / min, 1400 rpm / min, 1600 rpm / min, 1800 rpm / min, 2000 rpm / min or any two of these. By limiting parameters such as temperature, time, and rotation speed of the first mixing, it is ensured that the first packing material and the first solvent are in full contact, removing impurities from the surface of the first packing material and avoiding affecting its subsequent adsorption effect on N-nitrosovancomycin.

[0041] In some embodiments, the concentration of vancomycin hydrochloride in the first solution is 50,000–150,000 μg / mL, for example, 50,000 μg / mL, 60,000 μg / mL, 70,000 μg / mL, 80,000 μg / mL, 90,000 μg / mL, 100,000 μg / mL, 110,000 μg / mL, 120,000 μg / mL, 130,000 μg / mL, 140,000 μg / mL, 150,000 μg / mL, or any combination thereof. By limiting the concentration of vancomycin hydrochloride in the first solution, its compatibility with the removal method described above in this invention is improved, which helps to further improve the removal efficiency.

[0042] In some embodiments, the pH value of the second solution is 1.0 to 5.0, for example, a range consisting of 1.0, 2.0, 3.0, 4.0, 5.0, or any two thereof. By limiting the pH value of the second solution, it is essentially acidifying the vancomycin hydrochloride solution to be treated, which helps to improve the adsorption selectivity, making it more compatible with the removal method described above, and further improving the removal efficiency.

[0043] In some embodiments, the mass ratio of the first packing material to the vancomycin hydrochloride to be treated is (1:8) to (5:1), for example, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, or any combination thereof. By limiting the mass ratio of the first packing material to the vancomycin hydrochloride to be treated, sufficient first packing material is ensured for adsorbing N-nitrosovancomycin, while simultaneously ensuring removal efficiency.

[0044] In some embodiments, the temperature of the third mixing is 0–35°C, for example, a range of 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, or any combination thereof; the time is ≥0.5 h, preferably 0.5–6 h, for example, 0.5 h, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 5 h, 5.5 h, 6 h, or any combination thereof; and the rotation speed is 200–2000 rpm / min. By limiting the parameters such as temperature, time, and rotation speed of the third mixing, sufficient contact between the first solid phase and the second solution is ensured, allowing the first solid phase to adsorb N-nitrosovancomycin in the second solution to the maximum extent, thereby achieving the purpose of improving the removal effect.

[0045] This invention does not limit the specific method of pH adjustment and can be implemented using conventional methods in the art. For example, in some embodiments, an acid regulator is used to adjust the pH value of the first solution, that is, by adding an acid regulator to the first solution to make its pH value 1.0 to 5.0, thereby obtaining a second solution with a pH value of 1.0 to 5.0. The acid regulator is selected from at least one of formic acid, acetic acid, trifluoroacetic acid, hydrochloric acid, phosphoric acid, and nitric acid. In this way, while adjusting the pH value, compounds that affect the properties of vancomycin hydrochloride are avoided.

[0046] To obtain the final purified vancomycin hydrochloride product, the present invention further includes a purification and dehydration treatment of the vancomycin hydrochloride product solution to obtain the vancomycin hydrochloride product. The purification and dehydration treatment can be carried out using conventional methods in the art, such as freeze drying, spray drying, or crystallization drying, specifically in a freeze dryer or a spray dryer.

[0047] This invention also includes a regeneration process for the packing material after adsorption. Specifically, the second solid-liquid mixture is subjected to a second filtration to obtain a second solid phase and a second liquid phase. The second liquid phase is the vancomycin hydrochloride product solution, and the second solid phase is the first packing material adsorbed with N-nitrosovancomycin. The second solid phase is regenerated to allow for the recycling of the first packing material.

[0048] In the regeneration process, a regenerating agent is used to regenerate the second solid phase. The regenerating agent can be a mixed solution containing organic matter and an alkaline substance. The organic matter can be at least one of methanol, ethanol, acetonitrile, and acetone, and the alkaline substance can be sodium hydroxide. In the regenerating agent, the concentration of the organic matter is 50-85%, and the mass concentration of the alkaline substance is 0.2-1 mol / L. Preferably, an acetic acid solution or hydrochloric acid solution with a concentration of 0.2-1.0 mol / L is used as the acid adjuster.

[0049] The present application will be further described in detail below with reference to specific embodiments, which should not be construed as limiting the scope of protection claimed in the present application.

[0050] Example 1

[0051] Take 10g of Uni Sil C18 reversed-phase packing material and mix it with 100mL of 60% ethanol aqueous solution. Mix at 35℃ for 4h. After the first filtration, collect the solid phase to obtain the pretreated reversed-phase packing material.

[0052] 10g of vancomycin hydrochloride sample to be treated was mixed with 100mL of deionized water, and the pH was adjusted to 4.5 with acetic acid to obtain a vancomycin hydrochloride solution with a pH of 4.5 and a concentration of 100,000μg / mL.

[0053] The pretreated reversed-phase packing material and the vancomycin hydrochloride solution to be treated were mixed and mixed at 10°C for 3 hours. After a second filtration, the liquid phase was collected to obtain the vancomycin hydrochloride product solution.

[0054] Example 2

[0055] Take 5g of Uni Sil C18 reversed-phase packing material and mix it with 100mL of acetone aqueous solution with a volume concentration of 60%. Mix at 20℃ for 4h. After the first filtration, collect the solid phase to obtain the pretreated reversed-phase packing material.

[0056] 25g of vancomycin hydrochloride sample to be treated was mixed with 500mL of deionized water, and the pH value was adjusted to 3.0 with hydrochloric acid to obtain a vancomycin hydrochloride solution with a pH value of 3.0 and a concentration of 50000μg / mL.

[0057] The pretreated reversed-phase packing material and the vancomycin hydrochloride solution to be treated were mixed and mixed at 5°C for 3 hours. After a second filtration, the liquid phase was collected to obtain the vancomycin hydrochloride product solution.

[0058] Example 3

[0059] 36g of Uni Sil C18 reversed-phase packing material was mixed with 200mL of acetone aqueous solution with a volume concentration of 40% and mixed at 25℃ for 2h. After the first filtration, the solid phase was collected to obtain the pretreated reversed-phase packing material.

[0060] 12g of vancomycin hydrochloride sample to be treated was mixed with 100mL of deionized water, and the pH value was adjusted to 2.0 with hydrochloric acid to obtain a vancomycin hydrochloride solution with a pH value of 2.0 and a concentration of 120000μg / mL.

[0061] The pretreated reversed-phase packing material and the vancomycin hydrochloride solution to be treated were mixed and mixed at 15°C for 2 hours. After a second filtration, the liquid phase was collected to obtain the vancomycin hydrochloride product solution.

[0062] Example 4

[0063] Take 1g of Uni Sil C18 reversed-phase packing material and mix it with 50mL of 60% ethanol aqueous solution. Mix at 35℃ for 4h. After the first filtration, collect the solid phase to obtain the pretreated reversed-phase packing material.

[0064] 8g of vancomycin hydrochloride sample to be treated was mixed with 80mL of deionized water, and the pH was adjusted to 4.5 with acetic acid to obtain a vancomycin hydrochloride solution with a pH of 4.5 and a concentration of 100,000μg / mL.

[0065] The pretreated reversed-phase packing material and the vancomycin hydrochloride solution to be treated were mixed and mixed at 10°C for 3 hours. After a second filtration, the liquid phase was collected to obtain the vancomycin hydrochloride product solution.

[0066] Example 5

[0067] Take 50g of Uni Sil C18 reversed-phase packing material and mix it with 200mL of 60% ethanol aqueous solution. Mix at 35℃ for 4h. After the first filtration, collect the solid phase to obtain the pretreated reversed-phase packing material.

[0068] 10g of vancomycin hydrochloride sample to be treated was mixed with 100mL of deionized water, and the pH was adjusted to 4.5 with acetic acid to obtain a vancomycin hydrochloride solution with a pH of 4.5 and a concentration of 100,000μg / mL.

[0069] The pretreated reversed-phase packing material and the vancomycin hydrochloride solution to be treated were mixed and mixed at 10°C for 3 hours. After a second filtration, the liquid phase was collected to obtain the vancomycin hydrochloride product solution.

[0070] Example 6

[0071] The difference from Example 1 is that Uni Sil C18 is replaced with the reverse polymer filler UniPS 30-300. Specifically, the following steps are included:

[0072] Take 10g of UniPS 30-300 reversed polymer packing and mix it with 100mL of 60% ethanol aqueous solution. Mix at 35℃ for 4h. After the first filtration, collect the solid phase to obtain the pretreated reversed packing.

[0073] 10g of vancomycin hydrochloride sample to be treated was mixed with 100mL of deionized water, and the pH was adjusted to 4.5 with acetic acid to obtain a vancomycin hydrochloride solution with a pH of 4.5 and a concentration of 100,000μg / mL.

[0074] The pretreated reversed-phase packing material and the vancomycin hydrochloride solution to be treated were mixed and mixed at 10°C for 3 hours. After a second filtration, the liquid phase was collected to obtain the vancomycin hydrochloride product solution.

[0075] Example 7

[0076] The difference from Example 1 is that Uni Sil C18 is replaced with macroporous adsorption resin LSi-200. Specifically, the following steps are included:

[0077] Take 10g of macroporous adsorption resin and mix it with 100mL of 60% ethanol aqueous solution. Mix at 35℃ for 4h. After the first filtration, collect the solid phase to obtain the pretreated macroporous adsorption resin.

[0078] 10g of vancomycin hydrochloride sample to be treated was mixed with 100mL of deionized water, and the pH was adjusted to 4.5 with acetic acid to obtain a vancomycin hydrochloride solution with a pH of 4.5 and a concentration of 100,000μg / mL.

[0079] The pretreated macroporous adsorption resin and the vancomycin hydrochloride solution to be treated were mixed and mixed at 10°C for 3 hours. After a second filtration, the liquid phase was collected to obtain the vancomycin hydrochloride product solution.

[0080] Example 8

[0081] The difference from Example 1 is that "60% ethanol aqueous solution by volume" is replaced with "40% ethanol aqueous solution by volume".

[0082] Example 9

[0083] The difference from Example 1 is that "60% ethanol aqueous solution" is replaced with "80% ethanol aqueous solution".

[0084] Example 10

[0085] The difference from Example 1 is that "60% ethanol aqueous solution by volume" is replaced with "10% ethanol aqueous solution by volume".

[0086] Example 11

[0087] The difference from Example 1 is that "60% ethanol aqueous solution" is replaced with "90% ethanol aqueous solution".

[0088] Comparative Example 1

[0089] The difference from Example 1 is that the reversed-phase packing is not pretreated, and the reversed-phase packing is directly mixed with the vancomycin hydrochloride solution to be treated.

[0090] Comparative Example 2

[0091] The difference from Example 1 is that the pH of the vancomycin hydrochloride sample to be treated is not adjusted. Specifically, the following steps are included:

[0092] Take 10g of Uni Sil C18 reversed-phase packing material and mix it with 100mL of 60% ethanol aqueous solution. Mix at 35℃ for 4h. After the first filtration, collect the solid phase to obtain the pretreated reversed-phase packing material.

[0093] 10g of vancomycin hydrochloride sample to be treated was mixed with 100mL of deionized water to obtain a vancomycin hydrochloride solution with a concentration of 100000μg / mL.

[0094] The pretreated reversed-phase packing material and the vancomycin hydrochloride solution to be treated were mixed and mixed at 10°C for 3 hours. After a second filtration, the liquid phase was collected to obtain the vancomycin hydrochloride product solution.

[0095] Comparative Example 3

[0096] The difference from Example 1 is that the reversed-phase packing Uni Sil C18 is replaced with activated carbon.

[0097] Comparative Example 4

[0098] The difference from Example 1 is that the reverse phase packing Uni Sil C18 is replaced with the normal phase packing Uni Sil 10-100.

[0099] Test case

[0100] 1. Yield test of vancomycin hydrochloride product

[0101] Weigh the vancomycin hydrochloride sample to be treated, and record the mass as A. Dissolve the weighed vancomycin hydrochloride in deionized water at a concentration of 50,000–150,000 μg / mL to obtain the vancomycin hydrochloride solution to be treated. Mix the pretreated reversed-phase packing material and the vancomycin hydrochloride solution to be treated at a mass ratio of 1:8–2:1. After filtration, collect the liquid phase to obtain the vancomycin hydrochloride product solution. Record the concentration of vancomycin hydrochloride in the product solution as B, the volume of the product solution as C, and the mass of vancomycin hydrochloride in the product solution as D. The yield of the vancomycin hydrochloride product is shown below:

[0102] Vancomycin hydrochloride yield = (B × C) / A × 100% = D / A × 100%

[0103] 2. Test for N-nitrosovancomycin content

[0104] Analysis and detection of N-nitrosovancomycin in vancomycin hydrochloride raw material and formulations. This compound was standardized using a mass balance method. The specific chromatographic conditions for standardization are as follows:

[0105] Column: Waters ACQUITY CSHTM C18 150mm×2.1mm, 1.7μm;

[0106] Buffer solution: Weigh about 7.0 g of tris(hydroxymethyl)aminomethane, dissolve it in about 950 mL of water, adjust the pH to 8.0-8.3 with 20% glacial acetic acid solution, and dilute with water to 1000 mL.

[0107] Mobile phase A: a mixture of buffer solution, acetonitrile, and methanol in a volume ratio of 93:3:4; Mobile phase B: a mixture of buffer solution, acetonitrile, and methanol in a volume ratio of 50:10:40; Flow rate: 0.30 mL / min

[0108] Detection wavelength: 280nm

[0109] Injection volume: 3 μL

[0110] Column temperature: 40℃

[0111] Autosampler: 5℃

[0112] The gradient elution procedure is shown in Table 1 below:

[0113] Table 1

[0114] Time / min Mobile phase A / % Mobile phase B / % 0 88 12 7 88 12 21 75 25 35 25 75 37 25 75 38 88 12 45 88 12

[0115] The calculation formula is as follows:

[0116] Sample content (ppm) = (Cpair × Asupply) / Apair

[0117] In the formula:

[0118] C: Concentration of N-nitrosovancomycin in the reference solution, ng / mL;

[0119] Asupply: Peak area of ​​the test solution, mAU*s;

[0120] A: Peak area of ​​the reference solution, mAU*s;

[0121] The products in the examples and comparative examples were subjected to performance tests, and the test results are shown in Table 2.

[0122] Table 2

[0123]

[0124]

[0125] As shown in Table 2, the removal method provided by the present invention can reduce the content of N-nitrosovancomycin while ensuring the yield of vancomycin hydrochloride product.

[0126] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for removing N-nitroso-vancomycin from vancomycin hydrochloride, characterized by, The removal method comprises the following steps: mixing a first filler with a first solvent to obtain a first solid-liquid mixture; filtering the first solid-liquid mixture to collect a first solid phase; mixing the to-be-processed vancomycin hydrochloride with a second solvent to obtain a first solution; adjusting the pH value of the first solution to be acidic to obtain a second solution; mixing the first solid phase with the second solution to obtain a second solid-liquid mixture; filtering the second solid-liquid mixture to collect a liquid phase, which is a vancomycin hydrochloride product liquid; The first filler is a reverse-phase filler.

2. The removal method according to claim 1, characterized by, The first filler has a particle size of 2 to 200 μm and a pore size of 0.1 to 100 nm.

3. The removal method according to claim 1 or 2, characterized by, The first filler is selected from one or more of LSi-200, UniPS10-300, UniPS20-300, UniPS 30-300, UniSil C18, UniSil C8, and UniSil C4.

4. The removal method according to claim 1 or 2, characterized by, The first solvent is a mixture of an organic solvent and water, the organic solvent being selected from at least one of methanol, ethanol, and acetone; the second solvent is water; preferably, the volume concentration of the organic solvent in the first solvent is 40-80%.

5. The removal method according to claim 1 or 2, characterized by, The temperature of the first mixing is 0-35°C, the time is 0.5-6h, and the rotation speed is 200-2000rpm / min.

6. The removal method according to claim 1 or 2, characterized by, The concentration of vancomycin hydrochloride in the first solution is 50000-150000μg / mL.

7. The removal method according to claim 1 or 2, characterized by, The pH value of the second solution is 1.0-5.

0.

8. The removal method according to claim 1 or 2, characterized by, The mass ratio of the first filler to the to-be-processed vancomycin hydrochloride is (1:8)-(5:1).

9. The removal method according to claim 1 or 2, characterized by, The temperature of the third mixing is 0-35°C, the time is ≥0.5h, and the rotation speed is 200-2000rpm / min.

10. The removal method according to claim 1 or 2, characterized by, The pH of the first solution is adjusted by an acid regulator, wherein the acid regulator is selected from at least one of formic acid, acetic acid, trifluoroacetic acid, hydrochloric acid, phosphoric acid, and nitric acid; preferably, a solution of acetic acid or hydrochloric acid with a concentration of 0.2-1.0moL / L is used as the acid regulator.

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