A method for detecting related substances of cetirizine hydrochloride

Abstract

The present invention provides a method for detecting 2 chloroacetic acid residual amount in cetirizine hydrochloride, including: taking cetirizine hydrochloride to be tested and mixing with mobile phase, preparing a test solution; taking 2 chloroacetic acid sodium reference substance and mixing with mobile phase, preparing a reference solution; according to high performance liquid chromatography, using a chromatographic column with octadecylsilane bonded silica gel as a filler, the test solution and the reference solution are respectively injected into a high performance liquid chromatograph. The present invention can effectively detect 2 chloroacetic acid (salt) in cetirizine hydrochloride, meets the detection requirement for 2 chloroacetic acid (salt) residual amount in cetirizine hydrochloride, and is simple to operate, the result is stable and reliable, and the detection accuracy is high.

Description

Technical Field

[0001] The invention relates to a method for detecting the residual amount of 2-chloroacetic acid (salt) in cetirizine hydrochloride, and specifically relates to a method for detecting the residual amount of 2-chloroacetic acid (salt) in cetirizine hydrochloride by utilizing high performance liquid chromatography, and belongs to the field of drug analysis. Background Art

[0002] Cetirizine hydrochloride is a selective histamine H1 receptor antagonist. As a new antihistamine, it is mainly used to relieve allergic symptoms of allergic diseases. It is clinically used to treat allergic rhinitis, urticaria, angioedema and other skin and mucous membrane allergic diseases. It has many advantages such as rapid oral absorption, high receptor selectivity, strong clinical efficacy, few side effects and good safety. It is the first choice drug for the treatment of allergic diseases.

[0003] Cetirizine is a potent H1 receptor antagonist. It enters the body orally and quickly binds to the histamine H1 receptor on the target cell membrane, blocking histamine from activating the target cell. Cetirizine has no obvious anticholinergic and anti-5-hydroxytryptamine effects. At normal doses, it is not easy to penetrate the blood-brain barrier. This product can inhibit the transmission of histamine in the early stage of allergic reactions, reduce the migratory activity of inflammatory cells and the release of transmitters in the late stage of allergic reactions, so it is also effective for delayed allergic reactions. This product is a highly selective H1 receptor antagonist, and no adverse effects on cardiac electrical activity have been found so far.

[0004] 2-Chloroacetic acid is a reagent used in the production process of cetirizine hydrochloride raw materials. Existing technology confirms that 2-chloroacetic acid residues exist in drugs, and studies have found that the molecular structure of 2-chloroacetic acid indicates a warning structure and is on the list of Class 2B carcinogens. According to the ICH M7 guidelines, it is necessary to analyze and study the mutagenic impurities that may exist in cetirizine hydrochloride.

[0005] Patent 103263394B discloses a method for determining related substances in cetirizine hydrochloride tablets. The chromatographic conditions of the method are to use octadecylsilane bonded silica gel as a filler; acetonitrile-0.02% potassium dihydrogen phosphate solution (8:92) as mobile phase A (adjust the pH value to 3.0 with phosphoric acid), acetonitrile-0.05% phosphoric acid solution (48:52) as mobile phase B, and the flow rate is 1000 μg / min.

[0006] 1.0mL; column temperature is 40°C; detection wavelength is 230nm, and elution is performed in a linear gradient elution mode. This method is not specific for detecting the residual amount of 2-chloroacetic acid (salt) in cetirizine hydrochloride tablets, and the accuracy is not high. The gradient elution method has a poor retention effect on cetirizine hydrochloride, and the gradient elution is relatively cumbersome, and is easily affected by the purity of the mobile phase used in daily detection. In order to strictly require the quality of cetirizine hydrochloride, the present invention provides a method for detecting 2-chloroacetic acid (salt) in cetirizine hydrochloride with high sensitivity, strong specificity, stable analysis method, and simple operation, which effectively controls the quality of cetirizine hydrochloride. Summary of the invention

[0007] The invention adopts high performance liquid chromatography to detect 2-chloroacetic acid in cetirizine hydrochloride, and can be used for quality control in the production of cetirizine hydrochloride raw materials. The method has the characteristics of high sensitivity, strong specificity, good peak shape, and stable analysis method.

[0008] After continuous experiments, the following technical solutions were finally obtained:

[0009] A method for detecting the residual amount of 2-chloroacetic acid in cetirizine hydrochloride, the method using high performance liquid chromatography as a detection system, octadecylsilane bonded silica gel as a filler, methanol-acid solution as a mobile phase for isocratic elution, a detection wavelength of 200-242nm, and a column temperature of 20-40°C.

[0010] The mobile phase methanol can be replaced by one or both of isopropanol and acetonitrile.

[0011] The acid solution in the mobile phase is one of hydrochloric acid, sulfuric acid and acetic acid. Preferably, the acid solution in the mobile phase is acetic acid.

[0012] Furthermore, the pH value of the acidic aqueous solution in the mobile phase is 1.0-4.0.

[0013] Furthermore, the concentration of the acetic acid is 0.005 mol / L.

[0014] Furthermore, the flow rate of the mobile phase is 0.5-1.5 ml / min, preferably 0.8-1.2 ml / min.

[0015] Furthermore, the C18 column is an Agilent C18 chromatographic column or a Kromasil C18 chromatographic column.

[0016] Furthermore, the mobile phase is methanol-acetic acid aqueous solution, and the volume ratio is methanol: acetic acid aqueous solution = 55-67: 45-32.

[0017] Further, the high performance liquid chromatography method is:

[0018] Column temperature: 25°C;

[0019] The mobile phase was methanol: 0.005 mol / L acetic acid aqueous solution = 60:40;

[0020] The chromatographic column was Agilent C18 column;

[0021] The detection wavelength is 205nm;

[0022] Flow rate: 1.0 mL / min.

[0023] Furthermore, the cetirizine hydrochloride is a cetirizine hydrochloride bulk drug or preparation.

[0024] The invention discloses a method for detecting 2-chloroacetic acid in cetirizine hydrochloride by using high performance liquid chromatography. The analytical method can effectively detect the content of 2-chloroacetic acid in cetirizine hydrochloride, and the method has high separation degree, good repeatability and durability, low detection limit, simple operation, stable and reliable results, and can be used for quality control of cetirizine hydrochloride raw materials and preparations thereof, thereby providing effective guarantee for the quality of final products. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 is a chromatogram of a test solution according to an embodiment of the present invention.

[0026] Figure 2 is a chromatogram of a control solution according to an embodiment of the present invention.

[0027] Figure 3 is a chromatogram of a detection limit solution according to an embodiment of the present invention.

[0028] Figure 4 is a chromatogram of a quantitative limit solution according to an embodiment of the present invention.

[0029] Figure 5 is a chromatogram of a recovery solution according to an embodiment of the present invention. DETAILED DESCRIPTION

[0030] In order to make the purpose and technical scheme of the present invention clearer, the present invention is further described below in conjunction with the embodiments, but the protection scope of the present invention is not limited to these embodiments, and the embodiments are only used to explain the present invention. The mobile phase of the specific embodiment of the present invention is carried out with methanol, but isopropanol, acetonitrile or a mixture of 2 solvents can also achieve similar technical effects. Due to space limitations, no examples are given herein. Any changes or equivalent substitutions that do not deviate from the concept of the present invention are included in the protection scope of the present invention.

[0031] 1. Methodological research and evaluation

[0032] 1. Instruments and conditions:

[0033] Waters liquid chromatography system, 2998 detector, chromatographic column: Agilent SB-C18 (150×4.6mm, 5μm); detection wavelength: 205nm; methanol was used as mobile phase A, 0.005mol / L acetic acid water was used as mobile phase B, and the volume ratio was A:B=60:40 for isocratic elution. The column temperature was 25℃, the flow rate was 1.0ml / min, and the injection volume was 20μl.

[0034] 2. Test steps:

[0035] 2.1 System suitability experiment:

[0036] Accurately weigh an appropriate amount of cetirizine hydrochloride powder and use the mobile phase to make a solution containing about 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Measure according to the above chromatographic conditions and record the chromatogram. The results are as follows: Figure 1 shown.

[0037] It can be seen from the chromatogram that the retention time of 2-chloroacetic acid is 7.859 min, and the separation degree from the main peak of cetirizine hydrochloride and other impurity peaks meets the determination requirements.

[0038] Accurately weigh an appropriate amount of sodium 2-chloroacetate and use the mobile phase to prepare a solution containing about 4 μg per 1 ml as the system suitability solution. Measure according to the above-mentioned chromatographic conditions and record the chromatogram.

[0039] It can be seen from the chromatogram that the retention time of 2-chloroacetic acid is 7.859 min, and the separation degree from the main peak of cetirizine hydrochloride and other impurity peaks meets the determination requirements.

[0040] 2.2 Limit of quantification and limit of detection

[0041] 1) Detection limit: Accurately measure the reference solution and prepare a 0.080 μg / ml solution with the mobile phase. Determine it according to the law as the quantitative limit solution so that the signal-to-noise ratio (s / n) is greater than 3. The result is as follows: Figure 3 As shown in the chromatogram, the retention time of 2-chloroacetic acid is 7.375min.

[0042] 2) Limit of quantitation: Accurately measure the reference solution and prepare a 0.150 μg / ml solution with the mobile phase. Determine the solution as the limit of quantitation according to the law so that the signal-to-noise ratio (s / n) is greater than 10. The result is as follows: Figure 4 As shown in the chromatogram, the retention time of 2-chloroacetic acid is 7.367 min.

[0043] 2.3 Linearity and range

[0044] Take an appropriate amount of 2-chloroacetic acid reference substance and prepare solutions of concentrations of 0.150μg / ml, 0.300μg / ml, 1.200μg / ml, 3.600μg / ml, and 7.200μg / ml with water. Inject the solutions separately and measure according to the above-mentioned chromatographic conditions. Record the chromatogram and obtain the linear equation.

[0045] Table 1 Relationship between 2-chloroacetic acid peak area and concentration curve

[0046]

[0047] As shown in Table 1, the linear relationship between the concentration and the peak area of ​​2-chloroacetic acid is good within the range of 0.150-7.200 μg / ml.

[0048] 2.4 Repeatability Experiment

[0049] Take an appropriate amount of 2-chloroacetic acid reference substance and prepare a 4 μg / ml solution with mobile phase. Accurately measure 20 μl and inject it into the liquid chromatograph. Repeat the injection 6 times. Measure according to the above chromatographic conditions and record the chromatogram. The results are shown in Table 2.

[0050] Table 2 Repeated injection results of 2-chloroacetic acid reference substance

[0051]

[0052]

[0053] 2.5 Recovery rate experiment

[0054] Accurately weigh an appropriate amount of cetirizine hydrochloride powder, accurately weigh, add reference solution, add 2-chloroacetic acid, dissolve and dilute with mobile phase to prepare three concentrations of 80%, 100%, and 120% test samples, prepare three copies of each concentration in parallel, shake well, filter, and take the filtrate as the recovery solution. Determine according to the above chromatographic conditions and calculate the recovery rate. The results are shown in Table 3.

[0055] Table 3 2-Chloroacetic acid recovery rate

[0056]

[0057] 2.6 Durability test

[0058] 1) Waters liquid chromatography system, 2998 detector, chromatographic column: Agilent SB-C18 (150×4.6 mm, 5 μm); detection wavelength: 205 nm; methanol as mobile phase A, 0.005 mol / L acetic acid water as mobile phase B, volume ratio of A:B=55:45 for isocratic elution, column temperature of 25°C, flow rate of 1.0 ml / min, injection volume of 20 μl.

[0059] Determination: Take an appropriate amount of 2-chloroacetic acid reference substance, prepare it into a 4μg / ml solution with water, accurately measure 20μl and inject it into the liquid chromatograph, and record the chromatogram.

[0060] Under this condition, the retention time of 2-chloroacetic acid was about 7.698 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0061] 2) Waters liquid chromatography system, 2998 detector, chromatographic column: Agilent SB-C18 (150×4.6 mm, 5 μm); detection wavelength: 205 nm; methanol was used as mobile phase A, 0.005 mol / L acetic acid water was used as mobile phase B, and the volume ratio of A:B=67:33 was used for isocratic elution. The column temperature was 25°C, the flow rate was 1.0 ml / min, and the injection volume was 20 μl.

[0062] Determination: Take an appropriate amount of 2-chloroacetic acid reference substance, prepare it into a 4μg / ml solution with water, accurately measure 20μl and inject it into the liquid chromatograph, and record the chromatogram.

[0063] Under this condition, the retention time of 2-chloroacetic acid was about 8.056 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0064] 3) Waters liquid chromatography system, 2998 detector, chromatographic column: Agilent SB-C18 (150×4.6 mm, 5 μm); detection wavelength: 205 nm; methanol as mobile phase A, 0.005 mol / L acetic acid water as mobile phase B, volume ratio of A:B=62:38 for isocratic elution, column temperature of 20°C, flow rate of 1.0 ml / min, injection volume of 20 μl.

[0065] Determination: Take an appropriate amount of 2-chloroacetic acid reference substance, prepare it into a 4μg / ml solution with water, accurately measure 20μl and inject it into the liquid chromatograph, and record the chromatogram.

[0066] Under this condition, the retention time of 2-chloroacetic acid was about 7.824 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0067] 4) Waters liquid chromatography system, 2998 detector, chromatographic column: Agilent SB-C18 (150×4.6 mm, 5 μm); detection wavelength: 205 nm; methanol was used as mobile phase A, acetic acid water with pH=1.0 was used as mobile phase B, and the volume ratio of A:B=60:40 was used for isocratic elution. The column temperature was 25°C, the flow rate was 1.0 ml / min, and the injection volume was 20 μl.

[0068] Determination: Take an appropriate amount of 2-chloroacetic acid reference substance, prepare it into a 4μg / ml solution with water, accurately measure 20μl and inject it into the liquid chromatograph, and record the chromatogram.

[0069] Under this condition, the retention time of 2-chloroacetic acid was about 7.695 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0070] 5) Waters liquid chromatography system, 2998 detector, chromatographic column: Agilent SB-C18 (150×4.6 mm, 5 μm); detection wavelength: 205 nm; methanol was used as mobile phase A, acetic acid water with pH=3.0 was used as mobile phase B, and the volume ratio of A:B=60:40 was used for isocratic elution. The column temperature was 35°C, the flow rate was 1.0 ml / min, and the injection volume was 20 μl.

[0071] Determination: Take an appropriate amount of 2-chloroacetic acid reference substance, prepare it into a 4μg / ml solution with water, accurately measure 20μl and inject it into the liquid chromatograph, and record the chromatogram.

[0072] Under this condition, the retention time of 2-chloroacetic acid was about 7.354 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0073] 6) Waters liquid chromatography system, 2998 detector, chromatographic column: Agilent SB-C18 (150×4.6 mm, 5 μm); detection wavelength: 205 nm; methanol was used as mobile phase A, acetic acid water with pH=4.0 was used as mobile phase B, and the volume ratio of A:B=60:40 was used for isocratic elution. The column temperature was 40°C, the flow rate was 1.0 ml / min, and the injection volume was 20 μl.

[0074] Determination: Take an appropriate amount of 2-chloroacetic acid reference substance, prepare it into a 4μg / ml solution with water, accurately measure 20μl and inject it into the liquid chromatograph, and record the chromatogram.

[0075] Under this condition, the retention time of 2-chloroacetic acid was about 7.862 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0076] Embodiment 1: A method for detecting cetirizine hydrochloride 2-chloroacetic acid comprises the following steps:

[0077] 1) Instruments and conditions

[0078] Liquid chromatograph: Waters liquid chromatography system, 2998 detector;

[0079] Chromatographic column: Agilent SB-C18 (150×4.6mm, 5μm);

[0080] Chromatographic conditions: methanol as mobile phase A, acetic acid water with pH = 4.0 as mobile phase B, volume ratio of A:B = 60:40 for isocratic elution,

[0081] Detection wavelength: 205nm;

[0082] Column temperature: 40°C; flow rate: 1.0 ml / min; injection volume: 20 μl.

[0083] 1. Determination: Use appropriate amount of sodium 2-chloroacetate and make a solution containing about 4 μg per 1 ml with mobile phase as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0084] Under this condition, the retention time of 2-chloroacetic acid was about 7.862 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0085] 2. Determination: Accurately weigh an appropriate amount of cetirizine hydrochloride bulk drug fine powder and use the mobile phase to prepare a solution containing approximately 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0086] Under this condition, the retention time of 2-chloroacetic acid was about 7.398 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0087] 3. Determination: Accurately weigh an appropriate amount of cetirizine hydrochloride tablets and use the mobile phase to prepare a solution containing approximately 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0088] Under this condition, the retention time of 2-chloroacetic acid was about 8.002 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0089] Embodiment 2: A method for detecting cetirizine hydrochloride 2-chloroacetic acid comprises the following steps:

[0090] 1) Instruments and conditions

[0091] Liquid chromatograph: Waters liquid chromatography system, 2998 detector;

[0092] Chromatographic column: Agilent SB-C18 (150×4.6mm, 5μm);

[0093] Chromatographic conditions: isopropanol as mobile phase A, acetic acid water with pH = 4.0 as mobile phase B, volume ratio of A:B = 60:40 for isocratic elution;

[0094] Detection wavelength: 205nm;

[0095] Column temperature: 40°C; flow rate: 1.0 ml / min; injection volume: 20 μl.

[0096] 1. Determination: Use appropriate amount of sodium 2-chloroacetate and make a solution containing about 4 μg per 1 ml with mobile phase as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0097] Under this condition, the retention time of 2-chloroacetic acid was about 8.402 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0098] 2. Determination: Accurately weigh an appropriate amount of cetirizine hydrochloride bulk drug fine powder and use the mobile phase to prepare a solution containing approximately 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0099] Under this condition, the retention time of 2-chloroacetic acid was about 8.010 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0100] 3. Determination: Accurately weigh an appropriate amount of cetirizine hydrochloride tablets and use the mobile phase to prepare a solution containing approximately 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0101] Under this condition, the retention time of 2-chloroacetic acid was about 8.413 min, and there was no impurity peak that interfered with the detection of 2-chloroacetic acid.

[0102] Comparative Example 1

[0103] 1. Instruments and conditions

[0104] Liquid chromatograph: Waters liquid chromatography system, 2998 detector;

[0105] Chromatographic column: Agilent SB-C18 (150×4.6mm, 5μm);

[0106] Chromatographic conditions: methanol as mobile phase A, acetic acid water with pH = 3.0 as mobile phase B, volume ratio of A:B = 51:49 for isocratic elution,

[0107] Detection wavelength: 205nm;

[0108] Column temperature: 40°C; flow rate: 1.0 ml / min; injection volume: 20 μl.

[0109] 2. Determination: Use appropriate amount of sodium 2-chloroacetate and make a solution containing about 4 μg per 1 ml with mobile phase as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0110] Under this condition, no 2-chloroacetic acid peak was detected.

[0111] 3. Determination: Accurately weigh an appropriate amount of cetirizine hydrochloride bulk drug fine powder and use the mobile phase to prepare a solution containing approximately 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0112] Under this condition, no 2-chloroacetic acid peak was detected.

[0113] 4. Determination: Accurately weigh an appropriate amount of cetirizine hydrochloride tablets and use the mobile phase to prepare a solution containing approximately 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0114] Under this condition, no 2-chloroacetic acid peak was detected.

[0115] Comparative Example 2

[0116] 1. Instruments and conditions

[0117] Liquid chromatograph: Waters liquid chromatography system, 2998 detector;

[0118] Chromatographic column: Agilent SB-C18 (150×4.6mm, 5μm);

[0119] Chromatographic conditions: methanol as mobile phase A, acetic acid water with pH = 5.5 as mobile phase B, volume ratio of A:B = 60:40 for isocratic elution;

[0120] Detection wavelength: 205nm;

[0121] Column temperature: 40°C; flow rate: 1.0 ml / min; injection volume: 20 μl.

[0122] 2. Determination: Use appropriate amount of sodium 2-chloroacetate and make a solution containing about 4 μg per 1 ml with mobile phase as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0123] Under this condition, no 2-chloroacetic acid peak was detected.

[0124] 3. Determination: Accurately weigh an appropriate amount of cetirizine hydrochloride bulk drug fine powder and use the mobile phase to prepare a solution containing approximately 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0125] Under this condition, no 2-chloroacetic acid peak was detected.

[0126] 4. Determination: Accurately weigh an appropriate amount of cetirizine hydrochloride tablets and use the mobile phase to prepare a solution containing approximately 10 mg of cetirizine hydrochloride per 1 ml as the system suitability solution. Determine according to the above chromatographic conditions and record the chromatogram.

[0127] Under this condition, no 2-chloroacetic acid peak was detected.

[0128] From the above test results, it can be seen that the detection method of the present invention is scientific and reliable for detecting 2-chloroacetic acid in cetirizine hydrochloride raw materials and preparations thereof, the detection method is easy to operate, the separation degree of known impurities is high, the determination method has strong specificity, and the detection result is accurate and reliable. Therefore, the present method can be used for the quality control of cetirizine hydrochloride raw materials and preparations thereof.

Claims

1. A method for detecting the residual amount of 2-chloroacetic acid in cetirizine hydrochloride, characterized in that: The detection method is to use high performance liquid chromatography as a detection system, octadecylsilane bonded silica gel as a filler, and methanol-acid solution as a mobile phase for isocratic elution, wherein the pH value of the acidic aqueous solution in the mobile phase is 1.0-4.0; the C18 column is an Agilent C18 chromatographic column or a Kromasil C18 chromatographic column; the mobile phase is methanol-acetic acid aqueous solution, and in a volume ratio, methanol: acetic acid aqueous solution = 55-67: 45-32, the detection wavelength is 200-242nm, and the column temperature is 20-40°C.

2. The method according to claim 1, characterized in that The concentration of the acetic acid is 0.005 mol / L.

3. The method according to claim 1, characterized in that The flow rate of the mobile phase is 0.8-1.2 ml / min.

4. The method according to claim 1, characterized in that The HPLC method is: Column temperature: 25°C; The mobile phase was methanol: 0.005 mol / L acetic acid aqueous solution = 60:40; The chromatographic column is Agilent C18 column; The detection wavelength is 205nm; Flow rate: 1.0 mL / min.

5. The method according to claim 1, characterized in that The cetirizine hydrochloride is a cetirizine hydrochloride bulk drug or preparation.

6. The method according to any one of claims 1 to 5, characterized in that: In the high performance liquid chromatography, the mobile phase is one or both of isopropanol and acetonitrile.

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