Method for detecting contents of cetirizine hydrochloride and bacteriostatic agent in cetirizine hydrochloride drops

Abstract

The invention belongs to the technical field of drug detection, and particularly discloses a method for detecting the content of cetirizine hydrochloride and a bacteriostatic agent in cetirizine hydrochloride drops, which comprises the following steps: S1, precisely weighing a cetirizine hydrochloride drop sample, adding a solvent for dissolving, and diluting to a constant volume to prepare a test solution; s2, precisely weighing a cetirizine hydrochloride reference substance, a methyl hydroxybenzoate reference substance and a propyl hydroxybenzoate reference substance, adding a solvent for dissolving, and diluting to a constant volume to prepare a reference substance solution; s3, precisely measuring the test solution and the reference solution, respectively injecting the test solution and the reference solution into a high performance liquid chromatograph, and recording chromatograms; and calculating the contents of cetirizine hydrochloride, methylparaben and propylparaben by peak areas according to an external standard method. According to the method provided by the invention, cetirizine hydrochloride can be effectively separated from the bacteriostatic agent, and the requirement on quality control of cetirizine hydrochloride drops is met.

Description

Technical Field

[0001] The invention belongs to the technical field of drug detection, and particularly relates to a method for detecting the contents of cetirizine hydrochloride and an antibacterial agent in a cetirizine hydrochloride drop. Background Art

[0002] Cetirizine hydrochloride, chemical name: (±)-2-[2-[4-[(4-chlorophenyl)benzyl]-1-piperazinyl]ethoxy]acetic acid dihydrochloride, molecular weight 461.82, is a histamine H 1 Receptor antagonist, the structure is as follows:

[0003] Cetirizine hydrochloride has anti-allergic effects and is mainly used clinically to treat allergic diseases, such as seasonal rhinitis, perennial allergic rhinitis, allergic conjunctivitis, and itching and urticaria symptoms caused by allergies. Cetirizine hydrochloride has many advantages such as rapid oral absorption, high receptor selectivity, strong clinical efficacy, few side effects, and good safety. It has gradually become the first choice for the treatment of allergic diseases.

[0004] Drops are a common dosage form of cetirizine hydrochloride, in which antibacterial agents are added to ensure the quality stability of cetirizine hydrochloride drops within a specified time. However, excessive antibacterial agents can damage the patient's body. Therefore, the content of antibacterial agents must be strictly controlled to maintain the quality stability of the drops while avoiding harmful effects on patients. The antibacterial agents added to cetirizine hydrochloride drops include methylparaben and propylparaben, and their structures are as follows: Methylparaben: ; Propylparaben: .

[0005] At present, there is no universal method for detecting the content of antibacterial agents in cetirizine hydrochloride drops. Therefore, it is urgent to establish a reliable detection method to meet the needs of quality control of cetirizine hydrochloride drops. Summary of the invention

[0006] In view of the shortcomings of the prior art, the object of the present invention is to provide a method for detecting the content of cetirizine hydrochloride and antibacterial agent in cetirizine hydrochloride drops, which adopts high performance liquid chromatography with strong specificity, and can effectively separate the cetirizine hydrochloride peak, the antibacterial agent (including methylparaben and propylparaben) peak and the impurity peaks, thereby effectively solving the problems in the background technology.

[0007] The present invention is achieved through the following technical solutions: A method for detecting the content of cetirizine hydrochloride and antibacterial agent in cetirizine hydrochloride drops comprises the following steps: S1. Accurately weigh a sample of cetirizine hydrochloride drops, dissolve it in a solvent, dilute it to a fixed volume, and prepare a test solution; S2. Accurately weigh the cetirizine hydrochloride reference substance, methylparaben reference substance and propylparaben reference substance, dissolve them in solvent and dilute to a fixed volume to prepare a reference substance solution; S3. Accurately measure the test solution and the reference solution, inject them into the high performance liquid chromatograph respectively, and record the chromatogram; calculate the contents of cetirizine hydrochloride, methylparaben and propylparaben by the peak area according to the external standard method; Wherein, the antibacterial agent includes methylparaben and propylparaben, and the structure is as follows: Methylparaben: , Propylparaben: ; The solvent is 30% acetonitrile aqueous solution.

[0008] A further improvement of the present invention is: In S1, the concentration of cetirizine hydrochloride in the test solution is 0.08-0.12 mg / mL.

[0009] Furthermore, the concentration of cetirizine hydrochloride in the test solution is 0.10 mg / mL.

[0010] A further improvement of the present invention is: In S2, the concentration of cetirizine hydrochloride in the reference solution is 0.08-0.12 mg / mL, the concentration of methylparaben is 13.0-14.0 μg / mL, and the concentration of propylparaben is 1.2-1.8 μg / mL.

[0011] Furthermore, in the reference solution, the concentration of cetirizine hydrochloride is 0.10 mg / mL, the concentration of methylparaben is 13.5 μg / mL, and the concentration of propylparaben is 1.5 μg / mL.

[0012] A further improvement of the present invention is: In S3, the chromatographic conditions of the high performance liquid chromatograph are as follows: Chromatographic column: octadecylsilane bonded silica as filler, 4.6mm×250mm, 5μm; Mobile phase: acetonitrile-potassium hydrogen phosphate solution, where the volume ratio of acetonitrile to potassium hydrogen phosphate solution is 40:60; Detection wavelength: 230nm; Flow rate: 1.0 mL / min; Column temperature: 30°C; Injection volume: 50 μL.

[0013] Furthermore, the concentration of the dipotassium hydrogen phosphate solution is 3.0-4.0 g / L, and phosphoric acid is used to adjust the pH to 3.5-3.7.

[0014] Furthermore, the concentration of the dipotassium hydrogen phosphate solution is 3.5 g / L, and phosphoric acid is used to adjust the pH to 3.5.

[0015] A further improvement of the present invention is: In S3, the peak order of the chromatogram of the reference solution is cetirizine hydrochloride peak, methylparaben peak, and propylparaben peak. Beneficial Effects

[0016] Compared with the prior art, the present invention has the following obvious advantages: The present invention discloses a method for detecting the content of cetirizine hydrochloride and antibacterial agent in cetirizine hydrochloride drops, and adopts high performance liquid chromatography to separate cetirizine hydrochloride from two antibacterial agents (methylparaben and propylparaben). Among them, the peak of cetirizine hydrochloride, the peak of methylparaben and the peak of propylparaben can be effectively separated, and other impurity peaks do not interfere with the determination of cetirizine hydrochloride, methylparaben and propylparaben. The running time of each needle is 18 minutes, and the content of cetirizine hydrochloride and antibacterial agent (methylparaben and propylparaben) in cetirizine hydrochloride drops can be detected quickly and accurately. It has been verified that the method provided by the present invention has good specificity, precision, accuracy and stability, and can meet the quality control requirements of cetirizine hydrochloride drops. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the blank solvent chromatogram in the specificity test of Example 1; Figure 2 This is the chromatogram of blank auxiliary material solution in the specificity test of Example 1; Figure 3 This is the chromatogram of the reference substance solution in the specificity test in Example 1; Figure 4 This is the chromatogram of the test solution in Example 1: Specificity test; Figure 5 This is the chromatogram of the spiked solution of the test sample in Example 1: Specificity test; Figure 6 For Example 3: Standard curve of cetirizine hydrochloride in linearity and range; Figure 7 For Example 3: Linearity and range of the standard curve of methylparaben; Figure 8 This is the standard curve for propylparaben in Example 3: Linearity and range. DETAILED DESCRIPTION

[0018] The instruments, equipment, chromatographic columns, samples, reference substances, reagents, and test solutions used in the present invention are as follows: A. Instruments and Equipment

[0019] B. Chromatographic column

[0020] C. Samples, reference substances, reagents, and test solutions The cetirizine hydrochloride drops sample and drops auxiliary materials used in the examples were homemade, with production batch number: 20240822; the reference substance was purchased from the China Food and Drug Inspection Institute; the remaining reagents and test solutions were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0021] The present invention is described in detail below in conjunction with embodiments.

[0022] Example 1: Specificity test (1) Blank solvent: acetonitrile-water (volume ratio: 30:70).

[0023] (2) Blank excipient solution: Accurately measure 1 mL of blank drop excipient (excluding cetirizine hydrochloride, methylparaben, and propylparaben) and place it in a 100 mL volumetric flask. Add solvent to dissolve and dilute to the scale, and shake well.

[0024] (3) Cetirizine hydrochloride standard solution: Take about 10 mg of cetirizine hydrochloride reference substance, weigh accurately, place in a 100 mL volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0025] (4) Methylparaben reference solution: Take about 13.5 mg of methylparaben reference substance, accurately weigh it, place it in a 10 mL volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and use it as the methylparaben reference substance stock solution; accurately measure 1 mL of methylparaben reference substance stock solution, place it in a 100 mL volumetric flask, dilute to the scale with solvent, and shake well.

[0026] (5) Propylparaben reference solution: Take about 15 mg of propylparaben reference substance, accurately weigh it, place it in a 100 mL volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and use it as the propylparaben reference substance stock solution; accurately measure 1 mL of propylparaben reference substance stock solution, place it in a 100 mL volumetric flask, dilute to the scale with solvent, and shake well.

[0027] (6) Other degradation impurities localization solutions: Degradation impurities include cetirizine glyceride, cetirizine propylene glycol ester 1 and cetirizine propylene glycol ester 2, with the following structures: Cetirizine Glyceride: ; Cetirizine Propylene Glycol Ester1: ; Cetirizine Propylene Glycol Ester 2: .

[0028] Cetirizine glyceride positioning solution: Take about 10 mg of cetirizine glyceride reference substance, accurately weigh it, place it in a 200 mL volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0029] Cetirizine propylene glycol ester 1 localization solution: Take about 10 mg of cetirizine propylene glycol ester 1 reference substance, accurately weigh it, put it in a 250 mL volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0030] Cetirizine propylene glycol ester 2 positioning solution: Take about 10 mg of cetirizine propylene glycol ester 2 reference substance, accurately weigh it, put it in a 250 mL volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0031] (7) Reference solution: Take about 10 mg of cetirizine hydrochloride reference substance, weigh it accurately, and place it in a 100 mL volumetric flask. Accurately add 1 mL each of methylparaben reference substance stock solution and propylparaben reference substance stock solution. Add solvent to dissolve and dilute to the scale, and shake well.

[0032] (8) Test solution: Accurately measure 1 mL of cetirizine hydrochloride drops and place in a 100 mL volumetric flask. Add solvent to dissolve and dilute to the mark, and shake well.

[0033] (9) Test sample spiked solution: Accurately measure 1 mL of cetirizine hydrochloride drops sample and place it in a 100 mL volumetric flask. Accurately add 1 mL each of methylparaben reference stock solution, propylparaben reference stock solution, cetirizine glyceride fixation solution, cetirizine propylene glycol ester 1 fixation solution and cetirizine propylene glycol ester 2 fixation solution. Dissolve with solvent and dilute to scale. Shake well.

[0034] According to the analytical method provided by the present invention, 50 μL of each of the above solutions is accurately measured and injected into a high performance liquid chromatograph to record the chromatogram. The blank solvent chromatogram is shown in FIG. Figure 1 , the chromatogram of blank excipient solution is shown in Figure 2 , the chromatogram of the reference solution is shown in Figure 3 , the chromatogram of the test solution is shown in Figure 4 , the chromatogram of the test sample spiked solution is shown in Figure 5 .

[0035] Conclusion: Blank solvent and blank excipient have no interference on sample determination; in the chromatogram of the test sample spiked solution, the order of peaks is cetirizine glyceride, cetirizine, methylparaben, cetirizine propylene glycol ester 1, cetirizine propylene glycol ester 2, and propylparaben, and the main component peak, the two antibacterial agent peaks and the degradation impurity peaks can be effectively separated; the analytical method provided by the present invention has good specificity.

[0036] Example 2: Precision Test 1. Repeatability The preparation of the reference solution is the same as that in Example 1 under the specificity item.

[0037] Test solution: Accurately measure 1 mL of cetirizine hydrochloride drops sample, place in a 100 mL volumetric flask, add solvent to dissolve and dilute to the scale, shake well (prepare 6 portions in parallel).

[0038] According to the analytical method provided by the present invention, 50 μL of each of the above solutions was accurately measured and injected into a high performance liquid chromatograph, and the chromatogram was recorded. The RSD of the contents of methylparaben, cetirizine hydrochloride and propylparaben in 6 test solutions was determined. The results are shown in Table 1.

[0039] Table 1. Repeatability test results

[0040] Conclusion: The analytical method provided by the present invention has good repeatability.

[0041] 2. Intermediate precision The preparation of the reference solution is the same as that in Example 1 under the specificity item.

[0042] Test solution: Accurately measure 1 mL of cetirizine hydrochloride drops sample, place in a 100 mL volumetric flask, add solvent to dissolve and dilute to the scale, shake well (prepare 6 portions in parallel).

[0043] Different personnel, on different dates, using different instruments, followed the analytical method provided by the present invention, accurately measured 50 μL of each of the above solutions and injected them into a high performance liquid chromatograph, recorded the chromatograms, and measured 6 test solutions, respectively. The RSDs of the contents of methylparaben, cetirizine hydrochloride, and propylparaben in 12 test solutions (including the solutions under the repeatability item of this embodiment) were examined. The results are shown in Table 2.

[0044] Table 2, Intermediate precision test results

[0045] Conclusion: The analytical method provided by the present invention has good intermediate precision.

[0046] Example 3: Linearity and Range The preparation of methylparaben reference substance stock solution and propylparaben reference substance stock solution is the same as that of Example 1 under the specificity item.

[0047] Linear stock solution: Take about 10 mg of cetirizine hydrochloride reference substance, accurately weigh it, put it in a 10 mL volumetric flask, accurately add 1 mL each of methylparaben reference substance stock solution and propylparaben reference substance stock solution, add solvent to dissolve and dilute to the scale, and shake well.

[0048] The preparation methods of linear 1 to linear 5 solutions of cetirizine hydrochloride, methylparaben and propylparaben are shown in Table 3 below.

[0049] Table 3, Linear solution preparation table

[0050] According to the analytical method provided by the present invention, 50 μL of the linear 1 to linear 5 solutions of cetirizine hydrochloride, methylparaben and propylparaben prepared above were injected into a high performance liquid chromatograph, and the chromatogram was recorded. The standard curves were drawn with the concentration as the abscissa and the peak area as the ordinate, and the linear regression equation was calculated. The results are shown in Tables 4 to 6, and the standard curves are shown in Tables 4 to 6. Figure 6 to Figure 8 .

[0051] Table 4, Cetirizine hydrochloride linear determination results

[0052] Conclusion: Cetirizine hydrochloride has a good linear relationship in the concentration range of 52.4449μg / mL to 157.3347μg / mL. The linear regression equation is: y=62055x+31752, r=1.

[0053] Table 5, linear determination results of methylparaben

[0054] Conclusion: Methylparaben has a good linear relationship in the concentration range of 6.6300μg / mL to 19.8900μg / mL. The linear regression equation is: y=42545x+1711.2, r=1.

[0055] Table 6, Propylparaben linear determination results

[0056] Conclusion: The linear relationship of propylparaben is good in the concentration range of 0.7912μg / mL to 2.3736μg / mL. The linear regression equation is: y=35455x-241.99, r=0.9987.

[0057] Example 4: Accuracy Test The preparation of the reference solution is the same as that in Example 1 under the specificity item.

[0058] Cetirizine hydrochloride spiked stock solution: Take about 200 mg of cetirizine hydrochloride, accurately weigh it, place it in a 20 mL volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0059] Methylparaben spiked stock solution: Take about 27 mg of methylparaben reference substance, accurately weigh it, place it in a 20 mL volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0060] Propylparaben spiked stock solution: Take about 15 mg of propylparaben reference substance, accurately weigh it, place it in a 100 mL volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0061] 80% Test Solution: Accurately measure 1 mL of blank excipient, 0.8 mL of loratadine hydrochloride spiked stock solution, 0.8 mL of methylparaben spiked stock solution, and 0.8 mL of propylparaben spiked stock solution, transfer them into a 100 mL volumetric flask, dissolve with solvent and dilute to the mark, shake well (prepare 3 parallel samples).

[0062] 100% Test Solution: Accurately measure 1 mL of blank excipient, 1 mL of loratadine hydrochloride spiked stock solution, 1 mL of methylparaben spiked stock solution, and 1 mL of propylparaben spiked stock solution, transfer them into a 100 mL volumetric flask, dissolve with solvent and dilute to the mark, shake well (prepare 3 parallel samples).

[0063] 120% Test Solution: Accurately measure 1 mL of blank excipient, 1.2 mL of loratadine hydrochloride spiked stock solution, 1.2 mL of methylparaben spiked stock solution, and 1.2 mL of propylparaben spiked stock solution, transfer them into a 100 mL volumetric flask, dissolve with solvent and dilute to the mark, shake well (prepare 3 parallel samples).

[0064] According to the analytical method provided by the present invention, take 50 μL of each of the above solutions and inject them into a high performance liquid chromatograph respectively, record the chromatogram, and determine the recovery rates and RSDs of loratadine hydrochloride, methylparaben, and propylparaben in the test solutions at each concentration level. The results are shown in Tables 7 - 9.

[0065] Table 7, Results of the accuracy determination of loratadine hydrochloride

[0066] Table 8, Results of the accuracy determination of methylparaben

[0067] Table 9, Results of the accuracy determination of propylparaben

[0068] Conclusion: The analytical method provided by the present invention has good accuracy.

[0069] Example 5: Solution stability test The preparation of the reference solution and the test solution is the same as that under the specificity item of Example 1.

[0070] According to the analytical method provided by the present invention, take 50 μL of each of the above solutions and inject them into a high performance liquid chromatograph respectively, record the chromatogram, and examine the RSDs of the peak areas of methylparaben, loratadine hydrochloride, and propylparaben in the reference solution and the test solution within a certain period of time. The results are shown in Tables 10 - 11.

[0071] Table 10, Results of the stability determination of the reference solution

[0072] Table 11, Test solution stability test results

[0073] Conclusion: In the analytical method provided by the present invention, the reference solution and the test solution have good stability within 48 hours.

[0074] The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with the technology to understand the content of the present invention and implement it accordingly, and they cannot be used to limit the protection scope of the present invention. Any equivalent transformation or modification made according to the spirit of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for detecting the content of cetirizine hydrochloride and antibacterial agent in cetirizine hydrochloride drops, characterized in that: The steps include: S1. Accurately weigh a sample of cetirizine hydrochloride drops, dissolve it in a solvent, dilute it to a fixed volume, and prepare a test solution; S2. Accurately weigh the cetirizine hydrochloride reference substance, methylparaben reference substance and propylparaben reference substance, dissolve them in solvent and dilute to a fixed volume to prepare a reference substance solution; S3. Accurately measure the test solution and the reference solution, inject them into the high performance liquid chromatograph respectively, and record the chromatogram; calculate the contents of cetirizine hydrochloride, methylparaben and propylparaben by the peak area according to the external standard method; Wherein, the antibacterial agent includes methylparaben and propylparaben, and the structure is as follows: methylparaben: , Propylparaben: ; The solvent is 30% acetonitrile aqueous solution.

2. The method for detecting the content of cetirizine hydrochloride and antibacterial agent in a cetirizine hydrochloride drop according to claim 1, characterized in that: In S1, the concentration of cetirizine hydrochloride in the test solution is 0.08-0.12 mg / mL.

3. The method for detecting the content of cetirizine hydrochloride and antibacterial agent in a cetirizine hydrochloride drop according to claim 2, characterized in that: The concentration of cetirizine hydrochloride in the test solution is 0.10 mg / mL.

4. The method for detecting the content of cetirizine hydrochloride and antibacterial agent in a cetirizine hydrochloride drop according to claim 1, characterized in that: In S2, the concentration of cetirizine hydrochloride in the reference solution is 0.08-0.12 mg / mL, the concentration of methylparaben is 13.0-14.0 μg / mL, and the concentration of propylparaben is 1.2-1.8 μg / mL.

5. The method for detecting the content of cetirizine hydrochloride and antibacterial agent in a cetirizine hydrochloride drop according to claim 4, characterized in that: The concentration of cetirizine hydrochloride in the reference solution is 0.10 mg / mL, the concentration of methylparaben is 13.5 μg / mL, and the concentration of propylparaben is 1.5 μg / mL.

6. The method for detecting the content of cetirizine hydrochloride and antibacterial agent in a cetirizine hydrochloride drop according to claim 1, characterized in that: In S3, the chromatographic conditions of the high performance liquid chromatograph are as follows: Chromatographic column: octadecylsilane bonded silica as filler, 4.6mm×250mm, 5μm; Mobile phase: acetonitrile-potassium hydrogen phosphate solution, where the volume ratio of acetonitrile to potassium hydrogen phosphate solution is 40:60; Detection wavelength: 230nm; Flow rate: 1.0 mL / min; Column temperature: 30°C; Injection volume: 50 μL.

7. The method for detecting the content of cetirizine hydrochloride and antibacterial agent in cetirizine hydrochloride drops according to claim 6, characterized in that: The concentration of the dipotassium hydrogen phosphate solution is 3.0-4.0 g / L, and phosphoric acid is used to adjust the pH to 3.5-3.

7.

8. The method for detecting the content of cetirizine hydrochloride and antibacterial agent in cetirizine hydrochloride drops according to claim 7, characterized in that: The concentration of the dipotassium hydrogen phosphate solution is 3.5 g / L, and phosphoric acid is used to adjust the pH to 3.

5.

9. The method for detecting the content of cetirizine hydrochloride and antibacterial agent in a cetirizine hydrochloride drop according to claim 1, characterized in that: In S3, the peak order of the chromatogram of the reference solution is cetirizine hydrochloride peak, methylparaben peak, and propylparaben peak.

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