阅读说明：本技术 一种碘克沙醇的纯化方法 (Purification method of iodixanol ) 是由 江必旺 王希 吴永跃 陈强强 于 2019-01-16 设计创作，主要内容包括：本发明提供了一种碘克沙醇的纯化方法。本发明的碘克沙醇的纯化方法包括以下步骤：1)将碘克沙醇粗提取物进行溶解、过滤；2)将经步骤1)过滤后的碘克沙醇溶液上样到装有均粒反相硅胶的层析柱中进行层析；采用流动相对碘克沙醇溶液进行洗脱；3)分段收集目的峰值的碘克沙醇溶液,对符合要求的组份液进行汇总,得到纯化的碘克沙醇。本发明的碘克沙醇的纯化方法,采用均粒反相硅胶作为固定相,仅需一步层析纯化,即可做到碘克沙醇主要单杂小于0.1％、剩余单杂小于0.05％、纯度大于95％、收率大于88％,纯化收率高而稳定,同时本发明纯化方法简单方便,可用于规模化生产,大大降低生产成本。(The invention provides a purification method of iodixanol. The purification method of iodixanol comprises the following steps: 1) dissolving the crude iodixanol extract, and filtering; 2) loading the iodixanol solution filtered in the step 1) into a chromatographic column filled with homogeneous reversed-phase silica gel for chromatography; eluting with a mobile phase iodixanol solution; 3) and collecting iodixanol solution of a target peak value in a segmented manner, and summarizing the component liquid meeting the requirement to obtain the purified iodixanol. The purification method of iodixanol adopts homogeneous reversed phase silica gel as a stationary phase, only one-step chromatographic purification is needed, and the iodixanol has the main single impurity less than 0.1%, the residual single impurity less than 0.05%, the purity greater than 95%, the yield greater than 88%, and high and stable purification yield.)

1. A purification process for iodixanol, comprising the steps of:

1) dissolving the crude iodixanol extract, and filtering;

2) loading the iodixanol solution filtered in the step 1) into a chromatographic column filled with homogeneous reversed-phase silica gel for chromatography; eluting with a mobile phase iodixanol solution;

3) and collecting iodixanol solution of a target peak value in a segmented manner, and summarizing the component liquid meeting the requirement to obtain the purified iodixanol.

2. The purification method according to claim 1, wherein in the step 1), the purity of the iodixanol crude extract is 80-85%;

preferably, the filtration is to filter the iodixanol crude extract dissolved by pure water by using a filter membrane with the pore diameter of 0.4-0.5 mu m.

3. The purification method according to claim 1 or 2, wherein in step 2), the homogeneous reversed phase silica gel is UniSil C18.

4. The purification method according to any one of claims 1 to 3, wherein in step 2), the mobile phase is a mixture of ultrapure water and an organic solvent.

5. The purification method according to claim 4, wherein the mass concentration of the organic solvent in the mobile phase is 1 to 10%.

6. The purification method according to claim 4 or 5, wherein the organic solvent is any one or a mixture of at least two of methanol, ethanol, acetonitrile and isopropanol.

7. The purification method according to any one of claims 1 to 6, wherein in step 2), the elution is isocratic;

preferably, the isocratic elution is: and (3) washing and eluting by using an organic solvent-water solution with the mass concentration of the organic solvent being 2-5%, and after a peak appears in one column volume, washing and eluting by using an organic solvent-water solution with the mass concentration of the organic solvent being 6-9%.

8. The purification method according to any one of claims 1 to 7, wherein the step 2) further comprises a step of equilibration of the column prior to loading.

9. The purification method according to claim 8, wherein the balancing is performed by: removing impurities by using an organic phase-to-chromatography column with the mass concentration of 90-100%, and balancing the chromatography column after impurity removal by using an organic phase-to-water solution with the mass concentration of 2-5%;

preferably, the organic phase is any one of methanol, ethanol, acetonitrile and isopropanol or a mixture of at least two thereof.

10. Purification process according to one of claims 1 to 9, characterized in that it comprises the following steps:

1) dissolving a crude iodixanol extract with the purity of 80-85% by using pure water, and filtering the crude iodixanol extract dissolved by using the pure water by using a filter membrane with the pore diameter of 0.4-0.5 mu m;

2) loading the iodixanol solution filtered in the step 1) into a chromatographic column filled with homogeneous reversed-phase silica gel UniSil C18 for chromatography, removing impurities by using an organic phase-to-chromatographic column with the mass concentration of 90-100% before loading, and balancing the chromatographic column after removing the impurities by using an organic phase-to-water solution with the mass concentration of 2-5%;

performing isocratic elution on the iodixanol solution by using a mixture of ultrapure water and an organic solvent as a flowing phase, wherein the isocratic elution is as follows: washing and eluting with an organic solvent-water solution with the mass concentration of the organic solvent being 2-5%, and after a peak appears in a column volume, washing and eluting with an organic solvent-water solution with the mass concentration of the organic solvent being 6-9%;

3) and collecting iodixanol solution of a target peak value in a segmented manner, and summarizing the component liquid meeting the requirement to obtain the purified iodixanol.

Technical Field

The invention relates to the technical field of medicine purification, and particularly relates to a purification method of iodixanol.

Background

Iodixanol (iodixanol), an X-ray contrast agent. For adult angiography, cerebrovascular angiography (conventional with i.a.dsa), peripheral arterial angiography (conventional with i.a.dsa), abdominal angiography (i.a.dsa), urography, venography, and CT-enhanced examination. The structural formula is as follows:

at present, most of the reports of domestic and foreign patents, documents and the like on iodixanol are synthetic method patents, and a separation and purification method is only reported.

CN1273574A discloses a method for crystallizing iodixanol by using methanol/isopropanol/water, wherein the crystallization is performed under high heat energy, especially under the conditions of high pressure and temperature exceeding the boiling point of the solution, the conditions are harsh, and the requirements on production equipment are high.

CN101293855B discloses a purification method of iodixanol, which adopts a recrystallization method to purify a crude iodixanol product prepared by a method of dimerizing 5-acetamido-N, N' -bis (2, 3-dihydroxypropyl) -2, 4, 6-triiodoisophthalamide, and can obtain an iodixanol product suitable for pharmacopoeia standards through repeated purification for many times. The purification method can be completely applied to the purification of crude iodixanol prepared by a dimerization 5-acetamido-N, N' -bis (2, 3-dihydroxypropyl) -2, 4, 6-triiodoisophthalamide method, and the cost of the used solvent is lower, so that the cost can be obviously reduced compared with the existing purification method; however, the purification method of the invention is a non-chromatographic solvent purification method, and high purity and high content iodixanol can be obtained after repeated times.

Therefore, further research into the efficient purification of iodixanol is necessary.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a purification method of iodixanol, which has high and stable purification yield, is simple and convenient, can be used for large-scale production and greatly reduces the production cost.

In order to achieve the purpose, the invention adopts the following technical scheme:

a purification process for iodixanol comprising the steps of:

1) dissolving the crude iodixanol extract, and filtering;

2) loading the iodixanol solution filtered in the step 1) into a chromatographic column filled with homogeneous reversed-phase silica gel for chromatography; eluting with a mobile phase iodixanol solution;

3) and collecting iodixanol solution of a target peak value in a segmented manner, and summarizing the component liquid meeting the requirement to obtain the purified iodixanol.

The purification method of iodixanol adopts homogeneous reversed phase silica gel as a stationary phase, only one-step chromatographic purification is needed, and the iodixanol has the main single impurity less than 0.1%, the residual single impurity less than 0.05%, the purity greater than 95%, the yield greater than 88%, and high and stable purification yield.

In the step 1), the purity of the iodixanol crude extract is 80-85%, for example, the purity of the iodixanol crude extract is 80%, 81%, 82%, 83%, 84% or 85%.

In the step 1), the crude iodixanol extract dissolved in pure water is filtered with a filter membrane having a pore size of 0.4 to 0.5 μm, for example, the filter membrane has a pore size of 0.4 μm, 0.41 μm, 0.42 μm, 0.43 μm, 0.44 μm, 0.45 μm, 0.46 μm, 0.47 μm, 0.48 μm, 0.49 μm, 0.5 μm.

In the step 2), the homogeneous reversed phase silica gel is UniSil C18. UniSil C18 particle size of 30 μm and pore sizeThe filler has strictly controlled particle size and pore diameter structure, is monodisperse and has a pore channel structure, and has good pertinence when used as chromatographic filler.

In the step 2), the mobile phase is a mixture of ultrapure water and an organic solvent. The invention adopts organic solvent and ultrapure water as mobile phase, and can complete elution of iodixanol main peak by 14 column volumes, so the mobile phase used in the process is relatively safe and has lower cost.

Wherein the mass concentration of the organic solvent in the mobile phase is 1-10%, for example, the mass concentration of the organic solvent in the mobile phase is 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, etc.

The organic solvent is any one or a mixture of at least two of methanol, ethanol, acetonitrile and isopropanol. Typical but non-limiting combinations of the mixture are a mixture of methanol, ethanol, a mixture of methanol, acetonitrile, a mixture of methanol and isopropanol, a mixture of ethanol, acetonitrile, a mixture of ethanol and isopropanol, a mixture of acetonitrile and isopropanol, a mixture of methanol, ethanol, acetonitrile, a mixture of methanol, ethanol and isopropanol, a mixture of ethanol, acetonitrile and isopropanol, a mixture of methanol, ethanol, acetonitrile and isopropanol, and a mixture of methanol, ethanol, acetonitrile and isopropanol.

In the step 2), the elution is isocratic elution; preferably, the isocratic elution is: and (3) washing and eluting by using an organic solvent-water solution with the mass concentration of the organic solvent being 2-5%, and after a peak appears in one column volume, washing and eluting by using an organic solvent-water solution with the mass concentration of the organic solvent being 6-9%.

The step 2) also comprises a step of balancing the chromatographic column before loading.

The balancing process comprises the following steps: removing impurities by using an organic phase-to-chromatography column with the mass concentration of 90-100%, and balancing the chromatography column after impurity removal by using an organic phase-to-water solution with the mass concentration of 2-5%; for example, the mass concentration of the organic phase used for impurity removal is 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%; the mass concentration of the organic phase used for the balance is 2%, 3%, 4%, 5%, or the like.

The organic phase used for impurity removal and the organic phase used for equilibration-the organic phase in the aqueous solution may be the same or different. Preferably, the organic phase is any one or a mixture of at least two of methanol, ethanol, acetonitrile and isopropanol, typically but not limited to a combination of a mixture of methanol and ethanol, a mixture of methanol and acetonitrile, a mixture of methanol and isopropanol, a mixture of ethanol and acetonitrile, a mixture of ethanol and isopropanol, a mixture of acetonitrile and isopropanol, a mixture of methanol, ethanol and acetonitrile, a mixture of ethanol, acetonitrile and isopropanol, a mixture of methanol, ethanol, acetonitrile and isopropanol

As a preferred embodiment of the present invention, the purification method of iodixanol comprises the following steps:

1) dissolving the iodixanol crude extract with the purity of 80-85% by using pure water, and filtering the iodixanol crude extract dissolved by using the pure water by using a filter membrane with the pore diameter of 0.4-0.5 mu m;

2) loading the iodixanol solution filtered in the step 1) into a chromatographic column filled with homogeneous reversed-phase silica gel UniSil C18 for chromatography, removing impurities by using an organic phase-to-chromatographic column with the mass concentration of 90-100% before loading, and balancing the chromatographic column after removing the impurities by using an organic phase-to-water solution with the mass concentration of 2-5%;

performing isocratic elution on the iodixanol solution by using a mixture of ultrapure water and an organic solvent as a flowing phase, wherein the isocratic elution is as follows: washing and eluting with an organic solvent-water solution with the mass concentration of the organic solvent being 2-5%, and after a peak appears in a column volume, washing and eluting with an organic solvent-water solution with the mass concentration of the organic solvent being 6-9%;

3) and collecting iodixanol solution of a target peak value in a segmented manner, and summarizing the component liquid meeting the requirement to obtain the purified iodixanol.

Compared with the prior art, the invention has the beneficial effects that:

(1) the purification method of iodixanol adopts homogeneous reverse phase silica gel as a stationary phase, and only one-step chromatography is needed, so that the main single impurity of iodixanol is less than 0.1%, the residual single impurity is less than 0.05%, the purity is more than 95%, and the yield is more than 88%.

(2) The purification method of iodixanol provided by the invention has the advantages of high purity, high and stable yield, simple and convenient operation, reusability of the used stationary phase, relatively less mobile phase amount and greatly reduced cost.

Drawings

FIG. 1 is a high performance liquid chromatography analysis chart of a crude iodixanol extract before purification in example 1 of the present invention;

fig. 2 is a high performance liquid chromatography analysis diagram of iodixanol purified in example 1 of the present invention.

Detailed Description

The technical solution of the present invention is further described below with reference to fig. 1 and 2 by specific embodiments, but the present invention is not limited to these embodiments.