Method for detecting compound residues

Abstract

The invention discloses a compound residue detection method, which adopts liquid chromatography (HPLC) to perform quantitative analysis on residual vomitinib on the surface of equipment, can quickly and accurately determine the drug residue, and ensures that the production environment is clean and meets the drug production quality standard. The method comprises the following steps: preparing a solution, collecting a sample by using a wiping method or a leaching method, and preparing a diluent, a reference stock solution, a reference substance solution and a test solution. The chromatographic conditions are that a Thermo C18 chromatographic column is adopted, the detection wavelength is 332 nm, a mobile phase A is a phosphate buffer solution, a mobile phase B is acetonitrile, and the flow velocity is 1.0 ml / min. Through the method, the residual quantity of the vomitinib can be efficiently detected, the thorough cleaning of production equipment is ensured, the cross contamination of medicines is avoided, and the quality control level of the medicines is improved. The method has the advantages of simplicity and convenience in operation, rapidness in detection, high sensitivity and the like, and is widely applicable to detection and control of drug residues in the pharmaceutical industry.

Description

Technical Field

[0001] The invention belongs to the field of detection technology, and particularly relates to a method for detecting compound residues. Background Art

[0002] In the modern pharmaceutical production process, the problem of drug residues is an important link that cannot be ignored. With the continuous development of the pharmaceutical industry, especially the production of targeted drugs and biopharmaceutical products, the problem of drug residues in the production environment has become increasingly prominent. Drug residues not only affect the cleanliness of production equipment, but may also have a serious impact on the quality of subsequent production of drugs, and even pose a potential threat to the health of operators. Therefore, ensuring the thorough cleaning of production equipment and detecting and controlling drug residues have become key links in the quality management system of pharmaceutical companies.

[0003] Especially for highly active drugs such as anti-tumor drugs, the control of residual amount is particularly important. Vumetinib mesylate is abbreviated as Vumetinib; trade name: Aversa, which is a national Class 1 new drug independently developed by Shanghai Elis Pharmaceutical Technology Co., Ltd. It has become a highly selective and irreversible third-generation epidermal factor receptor tyrosine kinase inhibitor EGFR-TKI, used to treat disease progression during or after treatment with epidermal factor receptor inhibitor TKI, and confirmed by testing to have locally advanced or metastatic non-small cell lung cancer NSCLC with EGFR T790M mutation positive.

[0004] If the residues of vometinib are not effectively removed, it will not only affect the quality of subsequent drug production, but may also cause cross-contamination of drugs and may have adverse effects on the health of operators. Since vometinib itself has certain toxicity, excessive residues may cause environmental pollution and health risks.

[0005] Therefore, it is of great practical significance to develop an efficient, accurate and reliable detection method to scientifically and quantitatively detect drug residues in production equipment. Although existing detection methods can detect drug residues to a certain extent, most of them have disadvantages such as insufficient sensitivity, complex operation and long detection time, which make them difficult to be widely used in actual production processes. Summary of the invention

[0006] To solve the above problems, a compound residue detection method of the present invention can effectively detect the residual amount of vometinib in the production workshop and ensure that the cleanliness of the equipment meets the quality standards of drug production.

[0007] A method for detecting compound residues comprises the following steps:

[0008] S1: solution preparation;

[0009] S2: Liquid chromatography was used to analyze and determine the residual amount of afatinib mesylate on the production equipment in the workshop;

[0010] The solution prepared in step S1 includes a diluent, a blank solution, a control stock solution, a reference solution, and a test solution;

[0011] The chromatographic conditions in step S2 are:

[0012] Chromatographic column: Thermo C18 150mm×4.6mm×5μm;

[0013] Column temperature: 30°C;

[0014] Injection volume: 5 μl;

[0015] Detection wavelength: 332nm;

[0016] Flow rate: 1.0ml / min;

[0017] Running time: 5min;

[0018] Mobile phase: mobile phase A: mobile phase B = 50:50 (v / v);

[0019] Wherein, the mobile phase A is phosphate buffer, and the mobile phase B is acetonitrile.

[0020] In some embodiments, the solution preparation method is a wiping method, and the preparation method is:

[0021] Diluent: (0.7‰ dilute hydrochloric acid)-acetonitrile (25:75);

[0022] Blank solution: soak 4 cotton swabs in 0.1% glacial acetic acid solution, take them out and treat them like the sample;

[0023] Control stock solution: weigh 37.8 mg of vometinib mesylate reference substance, place in a 100 ml volumetric flask, add 80 ml of diluent, sonicate until completely dissolved, dilute to scale with diluent, shake well, prepare 2 portions in parallel, and use them as control stock solution A and reference substance stock solution respectively;

[0024] Reference solution: Accurately measure 2.0 ml of reference stock solution A and reference stock solution B into two 50 ml volumetric flasks, dilute to the mark with diluent, shake well, and use them as reference solution A and reference solution B respectively;

[0025] Test solution: Take 4 cotton swabs and wipe the sampling area 10cm×10cm=100cm 2 Each cotton swab selects 5cm×5cm=25cm 2, wipe 4 times vertically each time, then turn the cotton swab over and wipe 4 times horizontally, take the cotton swab after wiping, put it in a 20ml sampling bottle, accurately add 10.0ml of diluent, ultrasonicate for 20min, filter, and take the filtrate as the test solution.

[0026] In some embodiments, the solution preparation method is a leaching method, and the preparation method is:

[0027] Diluent: (0.7‰ dilute hydrochloric acid)-acetonitrile (25:75);

[0028] Blank solution: 0.1% glacial acetic acid for washing;

[0029] Control stock solution: weigh 37.8 mg of vometinib mesylate reference substance, place in a 100 ml volumetric flask, add 80 ml of diluent, sonicate until completely dissolved, dilute to scale with diluent, shake well, prepare 2 portions in parallel, respectively as control stock solution A and reference substance stock solution B;

[0030] Reference solution: Accurately measure 2.5 ml of reference stock solution A and reference stock solution B into two 50 ml volumetric bottles, dilute to the mark with diluent, shake well, then accurately measure 1.0 ml of the solution in the above two volumetric bottles into two 50 ml volumetric bottles, dilute to the mark with diluent, shake well, and use them as reference solution A and reference solution B respectively;

[0031] Test solution: Take the eluent, filter it, and take the filtrate as the test solution.

[0032] In some embodiments, system suitability confirmation is performed before detection, and the system suitability confirmation requirements are:

[0033] There should be no interference at the retention time of vometinib on the blank chromatogram;

[0034] 6. For reference solution A, the relative standard deviation (RSD) of the peak area of ​​vometinib peak is not greater than 10.0%;

[0035] The tailing factor of the main peak of the first reference solution A is between 0.8 and 1.8;

[0036] Prepare 1 portion of reference solution B, inject 1 needle, and calculate the recovery of the main peak to be 90.0% to 110.0% based on 6 needles of reference solution A;

[0037] The relative standard deviation (RSD) of the peak area of ​​the vometinib peak in the back-in control solution A and the previous 6 control solutions is not greater than 10.0%;

[0038] The peak area recovery of vometinib in reference solution B was calculated as follows:

[0039]

[0040] Where: As is the peak area of ​​the vometinib peak in reference solution B;

[0041] A R is the average peak area of ​​the vometinib peak in reference solution A in system suitability test 6;

[0042] W R When preparing reference solution A, weigh the amount of vometinib reference substance in mg;

[0043] W S When preparing reference solution B, weigh the amount of vometinib reference substance in mg.

[0044] In some embodiments, when the wiping method is used, the residual amount of vometinib is calculated according to the following formula:

[0045]

[0046] Where: As is the peak area of ​​vometinib in the test solution;

[0047] A R is the average peak area of ​​vometinib in reference solution A for system suitability 6;

[0048] W R The sample amount (mg) for preparing reference solution A;

[0049] f is the mass conservation content of the reference substance;

[0050] R is the accuracy (recovery) of the cleaning residue analysis method;

[0051] When 50%≤recovery rate<70%, R=0.5, and when 70%≤recovery rate≤130%, R=1.

[0052] In some embodiments, when the elution method is used, the residual amount of vometinib is calculated according to the following formula:

[0053]

[0054] Where: As is the peak area of ​​vometinib in the test solution;

[0055] A R is the average peak area of ​​vometinib in reference solution A for system suitability 6;

[0056] W R The sample amount (mg) for preparing reference solution A;

[0057] f is the mass conservation content of the reference substance;

[0058] R is the accuracy (recovery) of the cleaning residue analysis method.

[0059] In summary, the beneficial effects of the present invention are:

[0060] (1) The present invention uses high performance liquid chromatography (HPLC) technology, and can detect the residues of vometinib with high sensitivity and high accuracy through optimized chromatographic conditions (such as column temperature, flow rate, wavelength, etc.). Compared with traditional detection methods, HPLC technology can provide higher resolution and faster analysis speed, greatly improving the accuracy and efficiency of residue detection.

[0061] (2) The method provided by the present invention can be applied to the detection of the residual amount of vometinib on different equipment in the workshop, whether under the wiping method or the rinsing method. For the equipment surface that needs to be wiped and sampled, or the equipment that needs to be cleaned, the method can be used for effective detection to ensure the cleanliness of the production equipment and the quality of the drug. DETAILED DESCRIPTION

[0062] In order to deepen the understanding of the present invention, the present invention will be further described in detail below in conjunction with examples. The following examples are only used to explain the present invention and do not constitute a limitation on the scope of protection of the present invention.

[0063] A method for detecting the residual amount of afatinib mesylate on workshop production equipment.

[0064] First, prepare a high performance liquid chromatograph (HPLC): It is the core detection tool of this method, used to separate and quantitatively analyze the content of afatinib mesylate in the sample. Through the optimized chromatographic conditions, this method can ensure the rapid and accurate determination of vometinib. In this embodiment, the liquid chromatograph uses an Agilent 1260 HPLC system or other similar HPLC systems.

[0065] The specific chromatographic conditions are:

[0066] Chromatographic column: Thermo C18 150mm×4.6mm×5μm or similar column, which can effectively separate afatinib mesylate and other possible interfering substances and provide high-resolution separation effect.

[0067] Column temperature: 30 °C to ensure good separation and stability of vometinib.

[0068] Injection volume: 5 μl, to accommodate the standard analysis range.

[0069] Detection wavelength: 332nm. Afatinib mesylate has characteristic absorption at this wavelength, which can effectively improve the detection sensitivity.

[0070] Flow rate: 1.0ml / min, to ensure flow rate stability during chromatographic separation.

[0071] Running time: 5min;

[0072] Mobile phase: mobile phase A: mobile phase B = 50:50 (v / v), which can ensure the best separation effect of vometinib.

[0073] Mobile phase A: phosphate buffer (precisely weigh 2.7218 g of potassium dihydrogen phosphate, dissolve in 1000 ml of water, add 3 ml of triethylamine, adjust the pH to 4.0 with phosphoric acid solution (1→20) and filter);

[0074] Mobile phase B: acetonitrile.

[0075] The present invention provides two solution configuration methods: a wiping method and a rinsing method.

[0076] The preparation method of the wiping method is:

[0077] Diluent: (0.7‰ dilute hydrochloric acid)-acetonitrile (25:75);

[0078] Blank solution: soak 4 cotton swabs in 0.1% glacial acetic acid solution, take them out and treat them like the sample;

[0079] Control stock solution (323 μg / ml, based on vometinib): weigh 37.8 mg of vometinib mesylate reference substance (equivalent to 32.3 mg of vometinib), place in a 100 ml volumetric flask, add 80 ml of diluent, sonicate until completely dissolved, dilute to scale with diluent, shake well, prepare 2 portions in parallel, and use them as control stock solution A and reference substance stock solution respectively;

[0080] Reference solution (12.9 μg / ml, based on vometinib): accurately measure 2.0 ml of reference stock solution A and reference stock solution B into two 50 ml volumetric flasks, dilute to the mark with diluent, shake well, and use them as reference solution A and reference solution B respectively;

[0081] Test solution: Take 4 cotton swabs and wipe the sampling area 10cm×10cm=100cm 2 Each cotton swab selects 5cm×5cm=25cm 2 , make 4 S-shaped movements vertically each time, then turn the cotton swab over and wipe it 4 S-shaped movements horizontally. Place the wiping cotton swab in a 20ml sampling bottle, accurately add 10.0ml of diluent, ultrasonicate for 20min, filter, and take the filtrate as the test solution.

[0082] The preparation method of the elution method is:

[0083] Diluent: (0.7‰ dilute hydrochloric acid)-acetonitrile (25:75);

[0084] Blank solution: 0.1% glacial acetic acid for washing;

[0085] Control stock solution (323 μg / ml, based on vometinib): weigh 37.8 mg of vometinib mesylate reference substance (equivalent to 32.3 mg of vometinib), place in a 100 ml volumetric flask, add 80 ml of diluent, sonicate until completely dissolved, dilute to scale with diluent, shake well, prepare 2 portions in parallel, and use them as control stock solution A and reference substance stock solution B, respectively;

[0086] Reference solution (0.32 μg / ml, calculated as vometinib): accurately measure 2.5 ml of reference stock solution A and reference stock solution B into two 50 ml volumetric flasks, dilute to the mark with diluent, shake well, then accurately measure 1.0 ml of the solution in the above two volumetric flasks into two 50 ml volumetric flasks, dilute to the mark with diluent, shake well, and use them as reference solution A and reference solution B respectively;

[0087] Test solution: Take the eluent, filter it, and take the filtrate as the test solution.

[0088] When analyzing samples, inject according to the order in Table 1:

[0089]

[0090]

[0091] Table 1

[0092] System suitability confirmation is performed before actual analysis to ensure the stability and accuracy of the system when performing afatinib mesylate residue detection. The requirements for suitability confirmation include:

[0093] 1. There should be no interference at the retention time of vometinib on the blank chromatogram.

[0094] 2.6 For reference solution A, the relative standard deviation (RSD) of the peak area of ​​the vometinib peak was no greater than 10.0%.

[0095] 3. The tailing factor of the main peak of the first reference solution A is between 0.8 and 1.8

[0096] 4. Prepare 1 portion of reference solution B, inject 1 needle, and use 6 needles of reference solution A as the benchmark to calculate the recovery rate of the main peak, which is 90.0% to 110.0%.

[0097] 5. The relative standard deviation (RSD) of the peak area of ​​the vometinib peak in the returned control solution A and the previous 6 control solutions is not greater than 10.0%.

[0098] Among them, the peak area recovery rate of vometinib in reference solution B was calculated as follows:

[0099]

[0100] Where: As is the peak area of ​​the vometinib peak in reference solution B;

[0101] A R is the average peak area of ​​the vometinib peak in reference solution A in system suitability test 6;

[0102] W R When preparing reference solution A, weigh the amount of vometinib reference substance in mg;

[0103] W S When preparing reference solution B, weigh the amount of vometinib reference substance in mg.

[0104] After the test is completed, the residual amount of vometinib is calculated according to the following formula:

[0105] Calculation of residual amount by wiping method:

[0106]

[0107] Where: As is the peak area of ​​vometinib in the test solution;

[0108] A R is the average peak area of ​​vometinib in reference solution A for system suitability 6;

[0109] W R The sample amount (mg) for preparing reference solution A;

[0110] f is the mass conservation content of the reference substance;

[0111] R is the accuracy (recovery) of the cleaning residue analysis method;

[0112] Among them, when 50%≤recovery rate<70%, R=0.5, when 70%≤recovery rate≤130%, R=1.

[0113] Calculation of residual amount by elution method:

[0114]

[0115] Where: As is the peak area of ​​vometinib in the test solution;

[0116] A R is the average peak area of ​​vometinib in reference solution A for system suitability 6;

[0117] WR The sample amount (mg) for preparing reference solution A;

[0118] f is the mass conservation content of the reference substance;

[0119] R is the accuracy (recovery) of the cleaning residue analysis method.

[0120] Report the test results of each test solution. If it is less than the detection limit or no peak, report it as not detected; if it is less than the limit of quantification, report <LOQ (LOQ: specific value); if ≥LOQ, report it as the actual detection value.

[0121] In order to verify the accuracy and applicability of this method, the following projects were verified, which confirmed the accuracy and effectiveness of this method.

[0122] 1) Wiping method:

[0123]

[0124]

[0125]

[0126]

[0127] 2) Eluent method:

[0128]

[0129]

[0130]

[0131]

[0132] Through the detection method of the present application, we can effectively detect the residual amount of vometinib in the production workshop and ensure that the cleanliness of the equipment meets the quality standards of drug production. This detection method can not only improve the drug safety in the production process, but also prevent drug cross contamination and ensure the stability of drug quality. The method of the present invention is simple, accurate, and has a wide range of application prospects.

[0133] The above description shows and describes the preferred embodiments of the present invention. As mentioned above, it should be understood that the present invention is not limited to the form disclosed herein, and should not be regarded as excluding other embodiments, but can be used in various other combinations, modifications and environments, and can be modified within the scope of the invention concept described herein through the above teachings or the technology or knowledge of the relevant field. The changes and modifications made by those skilled in the art do not depart from the spirit and scope of the present invention, and should be within the scope of protection of the claims attached to the present invention.

Claims

1. A method for detecting compound residues, characterized in that: The following steps are involved: S1: solution preparation; S2: Liquid chromatography was used to analyze and determine the residual amount of afatinib mesylate on the production equipment in the workshop; The solution prepared in step S1 includes a diluent, a blank solution, a control stock solution, a reference solution, and a test solution; The chromatographic conditions in step S2 are: Chromatographic column: Thermo C18 150mm×4.6mm×5μm; Column temperature: 30°C; Injection volume: 5 μl; Detection wavelength: 332nm; Flow rate: 1.0ml / min; Running time: 5min; Mobile phase: mobile phase A: mobile phase B = 50:50 (v / v); Wherein, the mobile phase A is phosphate buffer, and the mobile phase B is acetonitrile.

2. The method for detecting compound residues according to claim 1, characterized in that: The solution preparation method is a wiping method, and the preparation method is: Diluent: (0.7‰ dilute hydrochloric acid)-acetonitrile (25:75); Blank solution: soak 4 cotton swabs in 0.1% glacial acetic acid solution, take them out and treat them like the sample; Control stock solution: weigh 37.8 mg of vometinib mesylate reference substance, place in a 100 ml volumetric flask, add 80 ml of diluent, sonicate until completely dissolved, dilute to scale with diluent, shake well, prepare 2 portions in parallel, and use them as control stock solution A and reference substance stock solution respectively; Reference solution: Accurately measure 2.0 ml of reference stock solution A and reference stock solution B into two 50 ml volumetric flasks, dilute to the mark with diluent, shake well, and use them as reference solution A and reference solution B respectively; Test solution: Take 4 cotton swabs and wipe the sampling area 10cm×10cm=100cm 2 Each cotton swab selects 5cm×5cm=25cm 2 , wipe 4 times vertically each time, then turn the cotton swab over and wipe 4 times horizontally, take the cotton swab after wiping, put it in a 20ml sampling bottle, accurately add 10.0ml of diluent, ultrasonicate for 20min, filter, and take the filtrate as the test solution.

3. The method for detecting compound residues according to claim 1, characterized in that: The solution preparation method is an elution method, and the preparation method is: Diluent: (0.7‰ dilute hydrochloric acid)-acetonitrile (25:75); Blank solution: 0.1% glacial acetic acid for washing; Control stock solution: weigh 37.8 mg of vometinib mesylate reference substance, place in a 100 ml volumetric flask, add 80 ml of diluent, sonicate until completely dissolved, dilute to scale with diluent, shake well, prepare 2 portions in parallel, respectively as control stock solution A and reference substance stock solution B; Reference solution: Accurately measure 2.5 ml of reference stock solution A and reference stock solution B into two 50 ml volumetric bottles, dilute to the mark with diluent, shake well, then accurately measure 1.0 ml of the solution in the above two volumetric bottles into two 50 ml volumetric bottles, dilute to the mark with diluent, shake well, and use them as reference solution A and reference solution B respectively; Test solution: Take the eluent, filter it, and take the filtrate as the test solution.

4. The method for detecting compound residues according to any one of claims 2 or 3, characterized in that: Before testing, the system suitability is confirmed. The system suitability confirmation requirements are as follows: There should be no interference at the retention time of vometinib on the blank chromatogram; 6. For reference solution A, the relative standard deviation (RSD) of the peak area of ​​vometinib peak is not greater than 10.0%; The tailing factor of the main peak of the first reference solution A is between 0.8 and 1.8; Prepare 1 portion of reference solution B, inject 1 needle, and calculate the recovery of the main peak to be 90.0% to 110.0% based on 6 needles of reference solution A; The relative standard deviation (RSD) of the peak area of ​​the vometinib peak in the back-in control solution A and the previous 6 control solutions is not greater than 10.0%; The peak area recovery of vometinib in reference solution B was calculated as follows: Where: As is the peak area of ​​the vometinib peak in reference solution B; A R is the average peak area of ​​the vometinib peak in reference solution A in system suitability test 6; W R When preparing reference solution A, weigh the amount of vometinib reference substance in mg; W S When preparing reference solution B, weigh the amount of vometinib reference substance in mg.

5. The method for detecting compound residues according to claim 4, characterized in that: When the wiping method is used, the residual amount of vometinib is calculated according to the following formula: Where: As is the peak area of ​​vometinib in the test solution; A R is the average peak area of ​​vometinib in reference solution A for system suitability 6; W R The sample amount (mg) for preparing reference solution A; f is the mass conservation content of the reference substance; R is the accuracy (recovery) of the cleaning residue analysis method; When 50%≤recovery rate<70%, R=0.5, and when 70%≤recovery rate≤130%, R=1.

6. The method for detecting compound residues according to claim 4, characterized in that: When the elution method is used, the residual amount of vometinib is calculated according to the following formula: Where: As is the peak area of ​​vometinib in the test solution; A R is the average peak area of ​​vometinib in reference solution A for system suitability 6; W R The sample amount (mg) for preparing reference solution A; f is the mass conservation content of the reference substance; R is the accuracy (recovery) of the cleaning residue analysis method.

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