CN-121994940-A - Starting material SM of ibutenib3Method for separating, identifying and measuring content of impurities thereof

Abstract

The invention belongs to the technical field of medicine analysis, and particularly relates to a method for separating, detecting and measuring an initial raw material SM 3 of ibutinib and impurities thereof, wherein the impurities comprise one or more of propionyl chloride, 3-chloropropionyl chloride and benzoyl chloride. The mobile phase is phosphate buffer solution as mobile phase A, acetonitrile as mobile phase B, octadecylsilane chemically bonded silica gel is adopted as filler in chromatographic column, and linear gradient elution of high performance liquid chromatography is carried out after pre-column derivatization. And the impurities are detected by a detector, and then the impurities are calculated according to the chromatogram by adopting an external standard method and the peak area. The method has the characteristics of good separation degree, good durability, high sensitivity and good reproducibility.

Inventors

• YANG YUFENG
• YU AO
• TAO NANNAN
• ZHANG RONG

Assignees

• 重庆华邦制药有限公司

Dates

Publication Date

20260508

Application Date

20241104

Claims (10)

1. 1. The method for separating and measuring the starting material SM 3 of the ibutinib and impurities thereof based on high performance liquid chromatography is characterized in that the starting material SM 3 of the ibutinib and the impurities jointly form a composition, wherein the impurities comprise any one or more of propionyl chloride, 3-chloropropionyl chloride and benzoyl chloride, the composition is subjected to pre-column derivatization to generate a derivative of the starting material SM 3 , propionyl chloride derivative, 3-chloropropionyl chloride derivative and benzoyl chloride derivative, the derivatization reagent is 1- (4-nitrophenyl) piperazine, the derivatized compound is separated by a high performance liquid phase, and the structural formula of the compound is as follows: the mobile phase is phosphate buffer solution as mobile phase A and acetonitrile as mobile phase B, and the stationary phase is chromatographic column with octadecylsilane chemically bonded silica gel as filler for linear gradient elution.
2. 2. The method according to claim 1, characterized in that the procedure of the linear gradient elution is as follows:
3. 3. The method according to claim 1, wherein the pre-column derivatization is specifically propionyl chloride, 3-chloropropionyl chloride, benzoyl chloride and 1- (4-nitrophenyl) piperazine heated at 40-60 ℃ for not less than 20 minutes to produce derivatives of starting material SM 3 , propionyl chloride derivatives, 3-chloropropionyl chloride derivatives, benzoyl chloride derivatives.
4. 4. The method of claim 1, wherein the starting volume ratio of mobile phase a to mobile phase B is 68-72:32-28 in the mobile phase.
5. 5. The method according to claim 1, wherein the flow rate is 1.0-1.2mL/min and the column temperature is 20-40 ℃.
6. 6. A method for identifying and measuring the starting material SM 3 of ibutinib and impurities thereof, which is characterized in that the composition is separated by adopting the method of any one of claims 1-5, and the composition is detected by a detector to obtain a chromatogram, and whether the starting material SM 3 for measuring ibutinib and impurities thereof are contained in the detected product is determined by comparing the chromatogram characteristics of the detected product and the reference product.
7. 7. The method of claim 6, wherein the detector has a detection wavelength of 228±10nm.
8. 8. The method according to claim 6, wherein the components and/or the derivatives corresponding to the components in the composition are identified in ascending order according to the order of the retention time, wherein the derivatives corresponding to the components are derivatives of the starting material SM 3 , the propionyl chloride derivative, the 3-chloropropionyl chloride derivative and the benzoyl chloride derivative.
9. 9. A method for determining the content of starting material SM 3 of ibutinib and impurities thereof, characterized in that the method according to any one of claims 5-8 is used for separating and identifying the starting material SM 3 of ibutinib and impurities thereof, obtaining a chromatogram, and calculating the impurities according to the external standard method and the peak area according to the obtained chromatogram.
10. 10. Use of said 1- (4-nitrophenyl) piperazine as a pre-column derivatizing agent for the composition of claim 1.

Description

Method for separating, identifying and measuring content of starting material SM 3 and impurities thereof of ibutinib Technical Field The invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for separating, detecting and determining an initial raw material SM 3 of ibutinib and impurities thereof. Background Ibutenib, also known as ibrutinib, is a Bruton's Tyrosine Kinase (BTK) inhibitor for the treatment of Chronic Lymphocytic Leukemia (CLL) and Mantle Cell Lymphoma (MCL). Seven impurities, namely acrylic anhydride, trichlorotoluene, acrylic acid, benzoic acid, propionyl chloride, 3-chloropropionyl chloride and benzoyl chloride, are commonly present in the synthesis process of ibrutinib. Wherein propionyl chloride, 3-chloropropionyl chloride and benzoyl chloride are genotoxic impurities, and a standard higher than the acceptable limit of the impurities in the raw materials (finished products) is formulated in the starting materials IBR-SM 3 according to the requirement of ICH M7 (R1) method 3. However, there is currently no method for detecting three genetic impurities. For example, gas phase analysis methods cannot be implemented. Because propionyl chloride, 3-chloropropionyl chloride, benzoyl chloride, the initial raw materials IBR-SM 3 and the possible impurity melting boiling points are lower, gas chromatography can be considered for measurement, but because the impurity is active and unstable, the impurity is easy to react with a gas chromatographic column stationary phase, the accuracy of sample detection is lost, and instrument equipment is easy to corrode, namely, propionyl chloride, 3-chloropropionyl chloride and benzoyl chloride cannot be quantitatively measured through gas chromatography, the accuracy of sample measurement cannot be ensured, gas-phase equipment such as a gas chromatographic column sampler and the like is easy to be damaged, the propionyl chloride, 3-chloropropionyl chloride, benzoyl chloride, the initial raw materials IBR-SM 3 and the possible impurity are strong in irritation and large in toxicity, the volatilized acyl chloride solvent gas is directly exposed to air in a gas direct measurement mode, and the injury to human body is large in the experimental process is a safety risk. At present, no literature report has been made on the use of a high performance liquid phase as a detection means. For example, CN111007157a discloses a method for detecting purity in the preparation process of ibrutinib, wherein before intermediate 003 is reacted with acryloyl chloride, the optical purity of ibrutinib intermediate is quantitatively analyzed by liquid chromatography, and the content of R-isomer is determined by liquid chromatography, so as to determine the purity of ibrutinib obtained by reacting R-isomer with the acryloyl chloride. However, this document does not disclose specific impurities, and the chromatographic conditions differ greatly from those of the present invention. For another example, CN104407067a discloses a liquid phase separation detection method of ibrutinib and enantiomer impurities thereof, belonging to the technical field of separation detection. The detection method comprises the steps of adopting AD-H,4.6mm X250mm and 5 mu m chiral chromatographic columns as separation chromatographic columns, and taking a mixed solvent of alkane solvent and alcohol solvent as a mobile phase, wherein the volume ratio of the alkane solvent to the alcohol solvent is 35:65. The method can effectively separate and detect ibrutinib and enantiomer impurities thereof. The impurities of the present invention are not disclosed and differ significantly from the chromatographic conditions of the present invention. At present, there is an urgent need to develop a method for detecting three genetic impurities. Disclosure of Invention It is an object of the present invention to provide a method for separating and assaying starting material SM 3 of ibutenib and impurities thereof, which can accomplish the separation of various substances in a short time. In order to achieve the above purpose, the technical scheme of the invention is as follows: The method for separating and measuring the starting material SM 3 of the ibutinib and impurities thereof based on a high performance liquid chromatography comprises the steps of forming a composition by the starting material SM 3 of the ibutinib and the impurities, wherein the impurities comprise any one or more of propionyl chloride, 3-chloropropionyl chloride and benzoyl chloride, derivatizing the composition before a column to generate a derivative of the starting material SM 3, propionyl chloride derivative, 3-chloropropionyl chloride derivative and benzoyl chloride derivative, the derivatizing reagent is 1- (4-nitrophenyl) piperazine, and separating the derivatized compound by using a high performance liquid phase, wherein the structural formula of the compound is shown in a table A. The separa