CN-121994984-A - Method for detecting content of 1-aminocyclopentylmethyl carbonitrile in 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride

Abstract

The invention provides a method for detecting the content of 1-aminocyclopentylmethylnitrile in 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride by adopting a high performance liquid chromatography, wherein chromatographic conditions are that a chromatographic column is adopted, namely YMC-Pack ODS-AQ,4.6mm×150mm and 5 mu m, an ultraviolet detector is adopted, the detection wavelength is 254nm, the column temperature is 25 ℃, the flow rate is 0.8ml/min, the sample injection volume is 50 mu l, gradient elution is adopted, the mobile phase A is a phosphoric acid aqueous solution with the concentration of 0.1%, and the mobile phase B is methanol.

Inventors

• WU TINGQIANG
• Zheng Qiuxing
• SONG HUI
• TANG WEIBIN
• CAI QIANG
• XIE LI

Assignees

• 珠海润都制药股份有限公司
• 润都制药(荆门)有限公司

Dates

Publication Date

20260508

Application Date

20241105

Claims (2)

1. 1. A method for detecting the content of 1-aminocyclopentylmethyl carbonitrile in 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride comprises the following steps: (1) Preparing a solution, namely preparing a reference substance of 1-aminocyclopentylmethylnitrile hydrochloride by using a 50% methanol-water solution to obtain a reference substance solution, preparing a sample to be tested of 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride by using a 50% methanol-water solution, and preparing a sample solution to be tested; (2) The sample solution and the reference substance solution are injected into a high performance liquid chromatograph for detection, the content of 1-aminocyclopentylmethylnitrile in the sample to be detected of the 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride is measured, the chromatographic conditions are that a chromatographic column is YMC-Pack ODS-AQ, the concentration is 4.6mm multiplied by 150mm, the chromatographic column is 5 mu m, an ultraviolet detector is adopted, the detection wavelength is 254nm, the column temperature is 25 ℃, the flow rate is 0.8ml/min, the injection volume is 50 mu l, gradient elution is adopted, the mobile phase A is phosphoric acid aqueous solution with the concentration of 0.1%, and the mobile phase B is methanol.
2. 2. The method for detecting the content of 1-aminocyclopentylmethylnitrile in 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride according to claim 1, wherein the gradient of the gradient elution is: 。

Description

Method for detecting content of 1-aminocyclopentylmethyl carbonitrile in 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride Technical Field The invention relates to the field of medicine analysis, in particular to a method for detecting the content of 1-aminocyclopentylmethylnitrile in 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride. Background 2-Butyl-1, 3-diazaspiro [4,4] non-1-en-4-one is a key synthetic intermediate for the preparation of the hypertension therapeutic drug irbesartan. In related studies it was found that 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one may be incorporated with the impurity 1-aminocyclopentylmethacrylonitrile and may remain in the final product irbesartan. According to ICH（The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use）'s guidelines, there are no detailed toxicological experimental data for many molecules, so the risk of the target molecule is generally estimated according to whether the target molecule contains a warning structural fragment, so as to determine the risk of impurities on drug safety. 1-aminocyclopentylmethyl carbonitrile is an impurity containing a genotoxicity warning structure, and the content of the impurity needs to be monitored and controlled to meet the safety requirement of a product. As a method for detecting impurities in irbesartan and intermediates thereof, a method for detecting impurities in irbesartan by using an LC-MS/MS method is reported in the article published in the volume 7 of the journal 59 of Chinese pharmacy, zhang Wenhui, et al, the related impurity structure is a series of molecules containing irbesartan parent nucleus skeleton, the chromatographic conditions are that a C-18 chromatographic column is adopted, a formic acid solution-acetonitrile (volume ratio of 62:38) with the volume fraction of 0.1% is adopted as a mobile phase, isocratic elution is carried out, liang Yanfen, et al, the article published in the volume 17 of food and medicine, and the volume 4 of the article are reported as a method for detecting impurities in irbesartan by using an HPLC method, wherein the impurity molecules are related substances A, and the separation conditions are that a C-18 chromatographic column is adopted, an acetonitrile-0.025 mol/L KH2PO4 solution (35:65, the pH is regulated to 6.0) is adopted as the mobile phase, and isocratic elution is carried out. It can be seen that in the current prior art, there is no relevant report about the detection of 1-aminocyclopentylmethane in irbesartan or intermediates thereof, and no relevant protocol is available. Therefore, there is a need to establish a method for detecting the content of 1-aminocyclopentylbutyronitrile in irbesartan or an intermediate thereof with sufficient precision and sensitivity to meet the quality monitoring requirements of pharmaceutical production. Disclosure of Invention Based on the technical background, the scheme of the invention provides a method for detecting the content of 1-aminocyclopentylmethylnitrile in 2-butyl-1, 3-diazaspiro [4,4] non-1-ene-4-one hydrochloride, the method has high detection sensitivity, and the system applicability, the specificity and the like of the method meet the requirements, and can meet the requirements of monitoring the production quality of medicines. The invention provides a method for detecting the content of 1-aminocyclopentylmethylnitrile in 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride which is an irbesartan synthesis intermediate, which comprises the following steps: (1) Preparing a solution, namely preparing a reference substance of 1-aminocyclopentylmethylnitrile hydrochloride by using a 50% methanol-water solution to obtain a reference substance solution, preparing a sample to be tested of 2-butyl-1, 3-diazaspiro [4,4] non-1-en-4-one hydrochloride by using a 50% methanol-water solution, and preparing a sample solution to be tested; (2) Sample the sample solution and the reference solution into a high performance liquid chromatograph under the chromatographic conditions of YMC-Pack ODS-AQ,4.6mm×150mm,5 μm, detection wavelength of 254nm with ultraviolet detector, column temperature of 25deg.C, flow rate of 0.8ml/min, sample volume of 50 μl, gradient elution, mobile phase A as 0.1% phosphoric acid aqueous solution, and mobile phase B as methanol. Further, the elution gradient of the gradient elution is: 。 a more specific operation flow is as follows: preparing a solution: a 50% aqueous methanol solution; Blank solution, namely diluent; 1-aminocyclopentylmethylnitrile stock solution 1-aminocyclopentylmethylnitrile hydrochloride reference substance is taken, precisely weighed, placed in a volumetric flask, dissolved and diluted to scale with diluent, and shaken well. (1-aminocyclopentylmethylnitrile hydrochloride: 666. Mu.g/ml, equivalent to 1-aminocyclopentylmethylnitrile: 500. Mu.g/ml) Precisely measuring 1.0ml of 1-aminocyclopentylmethylnitrile s