CN111751453A

CN111751453A - Analysis method of (3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidinamine salt

Info

Publication number: CN111751453A
Application number: CN201910247775.1A
Authority: CN
Inventors: 李敬芝; 徐静; 孙月静
Original assignee: Nanjing Gearpharma Technology Co ltd
Current assignee: Nanjing Gearpharma Technology Co ltd
Priority date: 2019-03-27
Filing date: 2019-03-27
Publication date: 2020-10-09

Abstract

The invention relates to a liquid chromatography analysis method of (3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidine amine hydrochloride isomer. Comprises the following steps: (1) dissolving acetyl Glucose Isothiocyanate (GITC) in acetonitrile to prepare a derivatization test solution; (2) precisely measuring the product and a derivatization test solution to prepare a test solution; (3) preparing a system adaptive solution; (4) the enantiomer of (3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidinamine hydrochloride was checked by high performance liquid chromatography to control the product quality. The method can scientifically, effectively and conveniently control the enantiomer of the compound by using a derivatization high performance liquid chromatography method, and has strong specificity and high sensitivity. The technical field of the method belongs to an analysis method of medicines.

Description

Analysis method of (3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidinamine salt

Technical Field

The invention relates to a liquid chromatography analysis method of (3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidine amine hydrochloride isomer, which has strong specificity and high sensitivity and has good effect on quality control. The technical field of the method belongs to an analysis method of medicines.

Background

(3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidine amine hydrochloride is an important starting material for synthesizing tofacitinib citrate. In order to produce the finished product tofacitinib citrate with excellent quality, the quality of the product needs to be strictly controlled.

The molecular structure of the product is as follows:

the product contains chiral carbon, and has diastereoisomer and enantiomer impurities, wherein the diastereoisomer impurities can be separated and detected by conventional related substance method, and the enantiomer is completely coincided with the main component under the related substance condition of common chromatographic column, so the separation is carried out by chiral chromatographic column or other methods. The product can be separated by a chiral chromatographic column (Chiralpak AGP), but the chromatographic column has low column efficiency, is limited to detect low impurity amount, and has short service life, thus being not beneficial to daily production monitoring. Therefore, after being derived, the product is measured by adopting a common C8 or C18 chromatographic column, and the method has the advantages of high sensitivity, strong specificity and good accuracy.

Disclosure of Invention

The invention provides a liquid chromatography analysis method for determining (3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidine amine hydrochloride enantiomer, which is characterized in that a derivatization high performance liquid chromatography method can scientifically, effectively and conveniently control the quality of the enantiomer.

The technical scheme of the invention is as follows:

1. preparation of derivatization test solution

An appropriate amount of acetylglucose isothiocyanate (GITC) was weighed out precisely, dissolved in acetonitrile and diluted to give a solution containing about 4.0mg per 1 ml.

2. Preparation of test solution

About 16mg of the product is precisely weighed, placed in a 20ml measuring flask, dissolved and diluted to the scale by adding a solvent (acetonitrile solution containing 0.2 percent of triethylamine), and shaken up to be used as a stock solution of a test article. Precisely measuring 1ml and 1ml of derivatization test solution, mixing uniformly, and standing for 40 minutes to obtain a test solution.

3. Preparation of System suitability solutions

Precisely weighing enantiomer about 2mg, placing in a 50ml measuring flask, adding a solvent to dissolve and dilute to scale, shaking up to obtain an impurity stock solution; precisely weighing 16mg of the product, placing the product in a 20ml measuring flask, adding 2ml of impurity stock solution, adding a solvent to dissolve and dilute the product to a scale, and shaking up to obtain a system applicability stock solution; precisely measuring 1ml of system applicability stock solution and 1ml of derivatization test solution, shaking up, and standing for 40 minutes to obtain the system applicability solution.

The suitable solvent in the step 1-3 is 0.1-0.3% of triethylamine in acetonitrile.

4. Measurement of

Subjecting the solution obtained in step 1-3 to chromatographic column with different polarities, gradient eluting at 20-40 deg.C and mobile phase flow rate of 0.8-1.0ml/min, and performing enantiomer measurement at 202-212 nm wavelength.

The chromatographic column with different polarities is a C18 column or a C8 column;

the organic solvent is acetonitrile;

the water phase is 0.01mol/L-0.05mol/L potassium dihydrogen phosphate solution (pH value is adjusted to 3.8 by phosphoric acid);

the gradient elution procedure is linear gradient elution.

The solvent is a mixed solvent of triethylamine and acetonitrile;

acetonitrile solution of triethylamine with the solvent proportion of 0.1 to 0.3 percent.

By adopting the high performance liquid chromatography condition, the enantiomer of the (3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidine amine hydrochloride can be effectively checked, so that the product quality can be controlled.

Drawings

FIG. 1 is a liquid chromatogram of an enantiomer

FIG. 2 is a liquid chromatogram of system applicability

FIG. 3 is a liquid chromatogram of a sample solution

FIG. 4 is a liquid chromatogram of a sample solution

FIG. 5 is a liquid chromatogram of a sample solution

FIG. 6 is a liquid chromatogram of a sample solution

FIG. 7 is a liquid chromatogram of a sample solution

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1

0.01mol/L potassium dihydrogen phosphate (pH value is adjusted to 3.8 by phosphoric acid) is taken as a mobile phase A; mobile phase B was acetonitrile. The chromatographic column is a C18 column; the detection wavelength is 207nm, and the column temperature is 30 ℃; the flow rate was 1.0 ml/min. The gradient elution procedure was as follows:

and precisely measuring 5 mu l of system applicability solution, injecting the solution into a liquid chromatograph, and recording a chromatogram. The separation degree of the main component and the enantiomer is required. And precisely measuring 5 mul of blank solution, enantiomer solution and test solution, injecting into a liquid chromatograph, and recording chromatogram. Calculated by area normalization (see FIGS. 1-3)

Example 2

0.03mol/L potassium dihydrogen phosphate (pH value is adjusted to 3.8 by phosphoric acid) is taken as a mobile phase A; mobile phase B was acetonitrile. The chromatographic column is a C8 column; the detection wavelength is 207nm, and the column temperature is 30 ℃; the flow rate was 1.0 ml/min. The gradient elution procedure was as follows:

precisely measuring 5 μ l of the test solution, injecting into a liquid chromatograph, and recording chromatogram. (see FIG. 4)

Example 3

0.01mol/L potassium dihydrogen phosphate (pH value is adjusted to 3.8 by phosphoric acid) is taken as a mobile phase A; mobile phase B was acetonitrile. The chromatographic column is a C18 column; the detection wavelength is 207nm, and the column temperature is 35 ℃; the flow rate was 0.8 ml/min. The gradient elution procedure was as follows:

precisely measuring 5 μ l of the test solution, injecting into a liquid chromatograph, and recording chromatogram. (see FIG. 5)

Example 4

0.02mol/L potassium dihydrogen phosphate (pH value is adjusted to 3.8 by phosphoric acid) is taken as a mobile phase A; mobile phase B was acetonitrile. The chromatographic column is a C8 column; the detection wavelength is 207nm, and the column temperature is 30 ℃; the flow rate was 0.9 ml/min. The gradient elution procedure was as follows:

precisely measuring 5 μ l of the test solution, injecting into a liquid chromatograph, and recording chromatogram. (see FIG. 6)

Example 5

0.01mol/L potassium dihydrogen phosphate (pH value is adjusted to 3.8 by phosphoric acid) is taken as a mobile phase A; mobile phase B was acetonitrile. The chromatographic column is a C18 column; the detection wavelength is 212nm, and the column temperature is 25 ℃; the flow rate was 1.0 ml/min. The gradient elution procedure was as follows:

precisely measuring 5 μ l of the test solution, injecting into a liquid chromatograph, and recording chromatogram. (see FIG. 7)

Claims

1. A liquid chromatography analysis method of tofacitinib citrate starting material (3R, 4R) -N, 4-dimethyl-1- (phenylmethyl) -3-piperidine amine hydrochloride is characterized in that the method can effectively perform optical isomer determination on the tofacitinib citrate starting material, and the technical scheme is as follows: using common derivatization reagent and chromatographic columns with different polarities, adopting a linear gradient program to elute under the detection conditions that the column temperature is 20-40 ℃ and the flow rate of a mobile phase is 0.8-1.0ml/min, and carrying out determination by using different wavelengths of an ultraviolet detector.

2. The liquid chromatography method of claim 1, wherein the different polarity column is a C18 column or a C8 column.

3. The liquid phase process of claim 1, wherein the mobile phase is a phosphate buffer and an organic solvent.

4. The liquid phase process of claim 1, wherein the solvent is a mixed solvent of triethylamine and acetonitrile.

5. The liquid chromatography method of claim 1, wherein said derivatizing agent is acetyl Glucose Isothiocyanate (GITC).

6. The liquid chromatography method of claim 1, wherein the detection wavelength is from 202nm to 212 nm.

7. The mobile phase according to claim 3, wherein the organic solvent is acetonitrile.

8. The mobile phase according to claim 3, wherein the aqueous phase is from 0.01mol/L to 0.05mol/L potassium dihydrogen phosphate (pH adjusted to 3.8 with phosphoric acid).

9. The solvent according to claim 4, characterized in that the solvent is a solution of 0.1-0.3% triethylamine in acetonitrile.