CN115575536A

CN115575536A - Method for measuring tartaric acid derivative in tartaric acid varenicline through high performance liquid chromatography

Info

Publication number: CN115575536A
Application number: CN202211227218.1A
Authority: CN
Inventors: 黄乐群; 周磊; 陆雨婷; 王磊
Original assignee: Jiangsu Sinobiopharma Co ltd
Current assignee: Jiangsu Sinobiopharma Co ltd
Priority date: 2022-10-09
Filing date: 2022-10-09
Publication date: 2023-01-06

Abstract

The invention provides a method for determining impurity J (monomethyl tartrate) and impurity K (dimethyl tartrate) in Vanilla tartrate by high performance liquid chromatography. The method can quantitatively detect the impurity J and the impurity K, thereby effectively controlling the content of the impurity, and having indispensable significance for the quality control of the varenicline tartrate.

Description

Method for measuring tartaric acid derivative in tartaric acid varenicline through high performance liquid chromatography

Technical Field

The invention relates to a method for determining tartaric acid derivatives in valnemulin tartrate by high performance liquid chromatography, belonging to the technical field of chemical detection and analysis.

Background

Vanillan is a non-nicotinic drug and is also a highly selective alpha 4 beta 2 neuronal nicotinic acetylcholine receptor partial agonist. The alpha 4 beta 2 nicotinic acetylcholine receptor is a part of nicotine combined with the brain, and can stimulate the nucleus accumbens of the brain to release a large amount of dopamine after the nicotine is combined with the nicotine, so that pleasure and nicotine reward effects are obtained; when the nicotine concentration is reduced, the pleasurable sensation disappears and the smoker will experience withdrawal symptoms. The main principle of the Vannit for smoking cessation is that alpha 4 beta 2 nicotinic acetylcholine receptors are partially excited, a small amount of dopamine is released, the craving of smokers for nicotine can be relieved, and the withdrawal symptoms are relieved; meanwhile, by antagonizing the combination of nicotine and alpha 4 beta 2 nicotinic acetylcholine receptor, the euphoria of smokers during smoking is blocked, and the smoking desire is reduced, so that the withdrawal rate is improved, and the relapse rate is reduced.

In 2006, month 5 and month 8, the U.S. Food and Drug Administration (FDA) and the european union medical agency (EMEA) successively approved a new non-nicotine smoking cessation drug, vareniclin tartrate, developed by fevery corporation for marketing. Approved for marketing in more than 100 countries, and is recommended by the latest edition of the guidelines for tobacco use and dependence on practice in the United states as the first line of treatment for smoking cessation. In 12 months of 2008, the pfeiy company announced that the drug is approved by the food and drug administration (SFDA) of China and is officially sold in the market of China (trade name: pei chang), and becomes the first smoking cessation prescription in China.

In the preparation process of the varenicline tartrate, an impurity J (monomethyl tartrate) and an impurity K (dimethyl tartrate) in a bulk drug are tartaric acid-derived impurities generated by tartaric acid inevitably contacting a solvent methanol in a synthesis process, and the impurities are controlled according to an ICH principle. Due to the esterification reaction after the tartaric acid is contacted with the methanol, the impurity J in the finished product is a main process impurity, the content measurement and control aiming at the impurity are one of the key points of the preparation process of the varenicline tartrate, and the quality of the finished product can be determined to a great extent. However, at present, there is no accurate quantitative detection method for the impurity J and the impurity K, which is not favorable for the complete establishment of the pharmaceutical quality control system of valnemulin tartrate.

Disclosure of Invention

The invention aims to fill the technical gap in the existing preparation and quality detection processes of a valnemulin tartrate drug, and provides a method for determining impurities J and K in valnemulin tartrate by using a high performance liquid chromatography.

The technical solution of the invention is as follows: a method for determining tartaric acid derivatives in Vanilla tartrate by high performance liquid chromatography comprises separating by ion exchange chromatography with weakly acidic solution as mobile phase system and chromatographic column with sulfonated crosslinked styrene and divinylbenzene copolymer as filler; the mobile phase is a weak acidic solution, wherein the weak acidic solution adopts an acidic reagent which comprises one or a mixture of more of formic acid, acetic acid, trifluoroacetic acid, phosphoric acid, citric acid and hydrochloric acid, and the mass concentration range is 0.05-0.2%.

The method specifically comprises the following steps:

1) Precisely weighing a to-be-detected varenicline tartrate product, placing the product in a measuring flask, adding a mobile phase to dissolve the product, and diluting the product into a solution containing 1.5mg of valenicline tartrate in 1mL of the solution to be used as a test solution;

2) Taking an appropriate amount of impurity J, impurity K and tartaric acid reference substance, precisely weighing, placing in a measuring flask, adding mobile phase for dissolving and diluting into a solution containing tartaric acid 0.6mg, impurity J7.5 μ g and impurity K1.5 μ g per 1ml, and using the solution as a system applicability solution;

3) Taking a proper amount of tartaric acid reference substances, precisely weighing, placing in a measuring flask, adding a mobile phase for dissolving and diluting into a solution containing about 0.6mg of tartaric acid per 1ml, precisely weighing 2ml, placing in a 100ml measuring flask, diluting to scale with the mobile phase, shaking up, precisely weighing 1ml, placing in a 10ml measuring flask, diluting to scale with the mobile phase, shaking up to serve as the tartaric acid reference solution;

4) And (3) high performance liquid chromatography detection: sulfonated cross-linked styrene and divinyl phenyl copolymer are used as a filling agent (such as ShodexRSpak KC-811 column or equivalent column), weak acid solution is used as a mobile phase, the detection wavelength is 220nm, and the column temperature is 40 ℃.

Precisely measuring 30 mu L of system applicability solution, injecting into a liquid chromatograph, recording a chromatogram of the system applicability solution, and ensuring that tartaric acid, impurity J and impurity K are in peak discharge in sequence; then precisely measuring 30 mu L of tartaric acid control solution, injecting the tartaric acid control solution into a liquid chromatograph, recording the chromatogram of the tartaric acid control solution, and adjusting the detection sensitivity to enable the peak height of the main component chromatographic peak to be 10-20% of the full range; and finally, precisely measuring the mobile phase, the sample solution and the tartaric acid control solution by 30 mu L respectively, injecting the mobile phase, the sample solution and the tartaric acid control solution into a liquid chromatograph respectively, and recording the chromatogram of the sample solution. After the solvent peak is deducted, if impurities J and K exist in a chromatogram of a sample solution, the content of the impurities J is not more than 0.50 percent calculated according to an external standard method; the content of the impurity K is not more than 0.10 percent according to the calculation of an external standard method added with a correction factor (multiplied by the correction factor of 1.3).

Compared with the prior art, the invention has the advantages that: the method can quantitatively detect the impurity J and the impurity K in the valnemulin tartrate medicine, strictly performs method verification through a large number of experiments, and ensures the scientificity and the rigidness of the method, thereby effectively controlling the content of the impurities, meeting the requirements of research and development and production, and having indispensable significance for the quality control of the valnemulin tartrate. From the aspects of improving the product quality, improving the safety of clinical medication and optimizing the process flow, the invention has more prominent commercial value, can form a technical barrier in enterprise standards and improve the added value of enterprise products.

Drawings

FIG. 1 is a typical chromatogram of the method for detecting impurity J and impurity K in the embodiment of the invention.

FIG. 2 is a typical chromatogram of a sample for detecting impurities J and K in the example of the present invention.

FIG. 3 is a typical chromatogram of blank solvent for detecting impurities J and K in the example of the invention.

Detailed Description

The technical solution of the present invention is further illustrated by the following examples. In the description of the present specification, the contents of each embodiment means that a specific technical feature described in connection with it is included in at least one embodiment of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the particular features described may be combined in any suitable manner in any one or more of the embodiments or examples.

Aiming at the synthesis process and degradation approach of the tartaric acid varenicline, the impurities which may appear in the esterification reaction between the tartaric acid and the methanol in the tartaric acid varenicline synthesis salt-forming process are summarized as shown in the following table 1:

TABLE 1 varenicline tartrate impurities related parameters

Separation is difficult to achieve due to its lack of retention under conventional reverse phase chromatography systems. The invention creatively searches a mobile phase system by inquiring documents and combining experimental experience, adopts the ion exchange chromatography to control the impurity J and the impurity K, adjusts the mobile phase proportion and the detection sensitivity, finally establishes the high performance liquid chromatography system and carries out methodology verification.

(1) Wavelength determination

And a diode array detector is adopted to carry out full scanning on the ultraviolet absorption wavelength of 190nm-400nm on each impurity, and the ultraviolet absorption curve of each substance is detected. Tartaric acid, impurity J and impurity K tend to be absorbed at the tail end, the product also has strong absorption near the tail end, the wavelength is temporarily 220nm as the detection wavelength, and tartaric acid and each impurity have strong absorption at the wavelength.

The method adopts ion exchange chromatography, takes weak acid solution as a mobile phase system, and selects a chromatographic column taking sulfonated cross-linked styrene and divinyl phenyl copolymer as a filler for separation.

(2) Detection concentration determination

Appropriate amounts of tartaric acid reference substance, impurity J and impurity K are taken to prepare system applicability solutions with detection concentrations of 1mg/ml, 1.5mg/ml and 2mg/ml, the contents of tartaric acid, impurity J and impurity K are respectively about 60%, 0.5% and 0.1%, each concentration of system applicability solution is injected with 30 mu L, 40 mu L and 50 mu L respectively, chromatographic behaviors of all components are synchronously inspected, and appropriate detection concentrations and injection amounts are selected. The sample concentrations and sample amounts were selected as shown in table 2 below:

TABLE 2 sample concentration and sample size selection

According to the results, when the detection concentration is 2mg/ml, the tartaric acid peak is wider, and the baseline fluctuation is larger; and the loss to the column in the later test of larger sample size will also increase. Compared with the prior art, the peak shapes of the peaks of the samples are symmetrical under the conditions that the concentration of the samples is 1.5mg/ml and the sample injection amount is 30 mu L, the base line is stable, and the separation effect is good; according to the sensitivity test, the result also fully meets the requirement of impurity detection sensitivity, and is suitable for accurately measuring the impurity J and the impurity K in the tartaric acid valnemulin.

(3) Methodology validation

The methodology of the method provided by the invention is verified according to the requirements of the Chinese pharmacopoeia 2015 edition. The results of the methodological verification of the relevant substances are as follows:

(1) specificity property

I. Destructive test

Acceptance criteria: the separation degree of each component of the degradation product is more than 1.5, and the peak purity of the main component is more than 0.999.

And (4) verification result: blank solvent is free of interference, and forced degradation tests show that the main component and each related substance peak can be completely separated, and the specificity is good; the peak purity of the main component peak and each known impurity peak meet the requirements, and the materials are basically kept in balance before and after damage. The results were in compliance with the regulations.

II. Degree of separation test

(2) Limit of quantification

Acceptance criteria: signal-to-noise ratio S/N =10.

And (4) verification result: calculated as signal-to-noise ratio S/N = 10:

tartaric acid: the quantitative limit is 0.31 mu g/ml, and the concentration ratio is 0.021%;

impurity J: the quantitative limit is 0.41 mu g/ml, and the concentration ratio is 0.027%;

impurity K: the quantitative limit is 0.20 mu g/ml, and the concentration ratio is 0.013%.

(3) Detection limit

Acceptance criteria: signal to noise ratio S/N =3.

And (4) verification result: calculated as signal-to-noise ratio S/N = 3:

tartaric acid: the quantitative limit is 0.10 mu g/ml, and the concentration ratio is 0.007%;

impurity J: the quantitative limit is 0.10 mu g/ml, and the concentration ratio is 0.007%;

impurity K: the quantitative limit is 0.08 mu g/ml, and the concentration ratio is 0.005%.

(4) Linearity and range

Acceptance criteria: the linear correlation coefficient R is more than 0.999.

And (4) verification result: tartaric acid was linear well in the range of 0.40 μ g/ml to 9.57 μ g/ml, R =0.9995;

impurity J is well linear in the range of 0.41 μ g/ml to 9.82 μ g/ml, R =0.9998;

impurity K was linear well in the range of 0.39 μ g/ml to 9.46 μ g/ml, R =0.9998.

(5) Correction factor

And (4) verification result: impurity J:1.0; impurity K:1.3.

(6) precision of sample introduction

Acceptance criteria: the relative standard deviation RSD is less than 2 percent.

And (4) verification result: the external standard method is used for continuously injecting the control solution for 6 times, and the RSD of the peak area is less than 2%.

(7) Repeatability of

Acceptance criteria: the relative standard deviation RSD is less than 2%.

And (4) verification result: 6 parts of test solution with the same concentration are prepared, and the RSD of the data of 6 parts of related substances is less than 2.0 percent.

(8) Accuracy of

Acceptance criteria: the recovery rate limit is 90-108%, and the relative standard deviation RSD is less than 3%.

And (4) verification result: recovery rate of impurity J: 96.01%, RSD:1.0 percent;

and (3) recovery rate of impurities K: 96.08%, RSD:6.1 percent.

(9) Durability

Acceptance criteria: small variations in the measurement conditions should be able to meet the system suitability test requirements.

And (4) verification result: the method inspects different detection conditions of the change of the flow rate relative value +/-10%, the change of the column temperature +/-5 ℃, the change of the concentration of an acid solution in a mobile phase and the change of the detection wavelength +/-2 nm, has trailing factors of main peaks smaller than 1.5, can separate all components well, and has good durability.

Stability of the solution in R

And (4) verification result: system applicability the samples and test solutions were stable under refrigerated (about 5 ℃) conditions for 11 days.

Verification results show that all indexes of the method meet the requirements of the 2015 edition of Chinese pharmacopoeia, and the method is suitable for accurately detecting the impurity J and the impurity K in the varenicline tartrate.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.

Furthermore, it should be understood that although the present specification describes embodiments, not every embodiment includes only a single embodiment, and such description is for clarity purposes only, and it is to be understood that all embodiments may be combined as appropriate by one of ordinary skill in the art to form other embodiments as will be apparent to those of skill in the art from the description herein.

Claims

1. A method for measuring tartaric acid derivatives in Vanilla tartrate by high performance liquid chromatography is characterized in that the method adopts ion exchange chromatography, takes weak acid solution as a mobile phase system, and specifically comprises the following steps:

2) Precisely weighing impurity J, impurity K and tartaric acid reference substance, placing in a measuring flask, adding mobile phase for dissolving and diluting into solution containing tartaric acid 0.6mg, impurity J7.5 μ g and impurity K1.5 μ g per 1ml, and using as system applicability solution;

3) Precisely weighing a tartaric acid reference substance, placing the tartaric acid reference substance into a measuring flask, adding a mobile phase to dissolve and dilute the tartaric acid reference substance into a solution containing 0.6mg of tartaric acid per 1ml, precisely weighing 2ml, placing the tartaric acid reference substance into a 100ml measuring flask, diluting the tartaric acid reference substance to a scale by using the mobile phase, shaking up, precisely weighing 1ml again, placing the tartaric acid reference substance into a 10ml measuring flask, diluting the tartaric acid reference substance to a scale by using the mobile phase, and shaking up to obtain a tartaric acid reference solution;

4) Precisely measuring 30 mu L of system applicability solution, injecting the system applicability solution into a liquid chromatograph, recording a chromatogram of the system applicability solution, and ensuring that tartaric acid, impurity J and impurity K are subjected to peak discharge in sequence; then precisely measuring 30 mu L of tartaric acid control solution, injecting the tartaric acid control solution into a liquid chromatograph, recording the chromatogram of the tartaric acid control solution, and adjusting the detection sensitivity to enable the peak height of the main component chromatographic peak to be 10-20% of the full range; and finally, precisely measuring the mobile phase, the sample solution and the tartaric acid control solution by 30 mu L respectively, injecting the mobile phase, the sample solution and the tartaric acid control solution into a liquid chromatograph respectively, and recording the chromatogram of the sample solution.

2. The method for measuring tartaric acid derivatives in valnemulin tartrate by high performance liquid chromatography as claimed in claim 1, wherein: the mobile phase is a weak acidic solution, wherein an acidic reagent adopted by the weak acidic solution comprises one or a mixture of more of formic acid, acetic acid, trifluoroacetic acid, phosphoric acid, citric acid and hydrochloric acid, and the mass concentration range is 0.05-0.2%.

3. The method for measuring tartaric acid derivatives in Vanilla tartrate by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: and 4) selecting sulfonated cross-linked styrene and divinylbenzene copolymer as filling agents for chromatographic columns of the liquid chromatograph in the step 4).

4. The method for measuring tartaric acid derivatives in Vanilla tartrate by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: the chromatographic conditions of the liquid chromatograph in the step 4) are as follows: the column temperature was 40 ℃ and the flow rate was 1.0 ml/min.

5. The method for measuring tartaric acid derivatives in Vanilla tartrate by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: the detector adopted by the liquid chromatograph in the step 4) is an ultraviolet detector.

6. The method for measuring tartaric acid derivatives in Vanilla tartrate by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: and in the step 4), the liquid chromatograph is eluted at equal degrees by adopting a reversed-phase high performance liquid chromatography.

7. The method for measuring tartaric acid derivatives in Vanilla tartrate by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: tartaric acid, the impurity J and the impurity K in the step 4) generate peaks in sequence, and the next step of operation is executed when the separation degree among the peaks is more than 1.5.

8. The method for measuring tartaric acid derivatives in valnemulin tartrate by high performance liquid chromatography as claimed in claim 1, wherein: after the solvent peak is deducted from the chromatogram of the sample solution in the step 4), if impurities J and K exist, the content of the impurities J is not more than 0.50 percent calculated according to an external standard method; the content of the impurity K is not more than 0.10 percent and the correction factor is 1.3 percent according to the calculation of an external standard method of adding the correction factor.