CN111983104A - Method for detecting content of chloroaniline and chlorhexidine gluconate in compound chlorhexidine gargle - Google Patents

Method for detecting content of chloroaniline and chlorhexidine gluconate in compound chlorhexidine gargle Download PDF

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CN111983104A
CN111983104A CN202010753810.XA CN202010753810A CN111983104A CN 111983104 A CN111983104 A CN 111983104A CN 202010753810 A CN202010753810 A CN 202010753810A CN 111983104 A CN111983104 A CN 111983104A
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chlorhexidine
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郎伟君
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Letai Pharmaceutical Co ltd
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Abstract

The invention discloses a method for detecting the content of chloroaniline and chlorhexidine gluconate in a compound chlorhexidine gargle. The compound chlorhexidine gargle is prepared from the following raw and auxiliary materials: 3000 parts of chlorhexidine gluconate solution, 500 parts of metronidazole 300, 50-150 parts of glycerol, 801-3 parts of polysorbate, 150 parts of polyethylene glycol 40050, 10-30 parts of concentrated peppermint water, 50-150 parts of ethanol, 300 parts of sodium saccharinate essence, 2-10 parts of lemon yellow, 1-5 parts of green tea essence and 2000 parts of purified water. In order to better control the product quality of the compound chlorhexidine gargle, the inventor researches a method for detecting the content of related substances, namely parachloroaniline and chlorhexidine gluconate, and provides a method for detecting the content of the parachloroaniline and the chlorhexidine gluconate in the compound chlorhexidine gargle.

Description

Method for detecting content of chloroaniline and chlorhexidine gluconate in compound chlorhexidine gargle
Technical Field
The invention relates to a medicine preparation method and a quality detection method thereof, in particular to a preparation method of a compound chlorhexidine gargle and a method for detecting the content of chloroaniline and glucose chlorhexidine in the compound chlorhexidine gargle, and belongs to the technical field of pharmacy.
Background
Oral diseases are common frequently encountered diseases and have high incidence rate. Common oral diseases include oral ulcer, periodontitis, gingivitis, toothache, loose teeth and the like. The compound chlorhexidine gargle is an over-the-counter medicine used in stomatology department, and is used for adjuvant treatment of gingival bleeding, periodontal swelling and pain, oral mucosa ulcer and the like caused by gingivitis, pericoronitis, oral mucositis and the like.
The chlorhexidine gluconate compound gargle contains chlorhexidine and metronidazole as effective components. In the preparation process of the existing chlorhexidine gluconate compound gargle, the generated impurities are mainly harmful substances such as parachloroaniline which can be absorbed by the undamaged skin, and the harmful substances are extremely harmful to human bodies. And the parachloroaniline cannot adopt a method for detecting related substances or content of the chlorhexidine gluconate in the existing batch quality standard, so that the methodological development needs to be carried out again. In addition, the existing preparation process of the chlorhexidine gluconate compound gargle can cause unstable product quality and generate precipitation in the storage process, thereby influencing the exertion of curative effect.
Therefore, the method for detecting the content of the chloroaniline and the glucose chlorhexidine in the compound chlorhexidine gargle is provided, and has important significance for controlling the quality of the compound chlorhexidine gargle.
Disclosure of Invention
One of the purposes of the invention is to provide a method for detecting the content of the chloroaniline and the chlorhexidine gluconate in the compound chlorhexidine gargle
The invention also aims to provide the compound chlorhexidine gargle and the preparation method thereof.
In order to achieve the purpose, the invention adopts the following technical means:
the invention relates to a method for detecting the content of chloroaniline and chlorhexidine gluconate in a compound chlorhexidine gargle, which comprises the following steps:
(1) preparation of test solution
Precisely measuring 5ml of compound chlorhexidine gargle to be measured, placing the gargle in a 100ml measuring flask, diluting the gargle to a scale with a diluent solvent, and shaking up to be used as a test solution; wherein the diluting solvent is prepared by the following method: 27.6g of sodium dihydrogen phosphate is weighed, 2000ml of water is added, and the pH is adjusted to 3.0 by phosphoric acid;
(2) preparation of reference stock solution
Accurately weighing 12mg of chlorhexidine acetate, placing the chlorhexidine acetate in a 50ml measuring flask, adding 5ml of acetonitrile and a proper amount of diluting solvent, dissolving by ultrasonic, diluting to scale with the diluting solvent, shaking up, and using the chlorhexidine acetate as a reference stock solution; accurately weighing 12mg of parachloroaniline, placing the parachloroaniline into a 100ml measuring flask, adding a proper amount of a diluting solvent, performing ultrasonic dissolution, diluting the parachloroaniline to a scale with the diluting solvent, and shaking up to be used as a parachloroaniline reference substance stock solution;
(3) preparation of Mixed control solutions
Precisely measuring the reference stock solution obtained in the step (2), diluting the reference stock solution into a mixed reference stock solution containing 42 mu g of chlorhexidine acetate and 0.12 mu g of parachloroaniline in each 1ml of diluted solvent, and shaking up to obtain a reference stock solution;
(4) liquid chromatograph detection
Precisely measuring the sample solution and the reference solution by 50 μ l each, injecting into a liquid chromatograph, and recording chromatogram; if a chromatographic peak with the same retention time as parachloroaniline in the chromatogram of the reference solution exists in the chromatogram of the test solution, calculating according to the peak area by an external standard method; the chlorhexidine gluconate is calculated by peak area according to an external standard method, and the result is multiplied by 1.4352 to obtain the chlorhexidine gluconate;
the calculation formula calculated by peak area according to an external standard method is as follows:
Figure BDA0002610865620000021
Figure BDA0002610865620000022
Cfor supplying to-the concentration of the component to be tested in the test sample; cTo pair-the concentration of the component to be measured in the control;
Afor supplying toPeak area of the component to be measured in the test sample; a. theTo pairPeak area of the component to be measured in the control;
Ssign boardThe marked amount of the product.
Among them, preferably, the detection conditions of the liquid chromatography in the step (4) are as follows: a sulfonic bonded silica gel chromatographic column is used as a filling agent, a phosphate solution is used as a mobile phase A, and acetonitrile is used as a mobile phase B; gradient elution was performed in the following order:
Figure BDA0002610865620000023
Figure BDA0002610865620000031
the detection wavelength is 239nm, and the column temperature is 40 ℃.
Wherein, preferably, the phosphate solution is prepared according to the following method: 27.6g of sodium dihydrogen phosphate was weighed, 10ml of triethylamine was added, 2000ml of water was added, and pH was adjusted to 3.0 with phosphoric acid.
Wherein, preferably, the chlorhexidine containing glucose should be 90.0% -110.0% of the marked amount; the content of p-chloroaniline must not exceed 0.2% of the limit.
Further, the invention also provides a compound chlorhexidine gargle which is prepared from the following raw and auxiliary materials in parts by weight: 3000 parts of chlorhexidine gluconate solution, 500 parts of metronidazole 300, 50-150 parts of glycerol, 801-3 parts of polysorbate, 150 parts of polyethylene glycol 40050, 10-30 parts of concentrated peppermint water, 50-150 parts of ethanol, 300 parts of sodium saccharinate essence, 2-10 parts of lemon yellow, 1-5 parts of green tea essence and 2000 parts of purified water.
Preferably, the compound chlorhexidine gargle consists of the following raw and auxiliary materials in parts by weight: 2400 parts of chlorhexidine gluconate solution, 400 parts of metronidazole, 100 parts of glycerol, 802 parts of polysorbate, 100 parts of polyethylene glycol 400, 20 parts of concentrated peppermint water, 110 parts of ethanol, 200 parts of sodium saccharinate, 6 parts of lemon yellow, 2 parts of green tea essence and 1650 parts of purified water.
Wherein, preferably, the concentrated peppermint water is prepared by the following method:
adding purified water into 12L of 90% ethanol, stirring, slowly adding 0.4L of dementholized peppermint oil, stirring, adding purified water to constant volume of 20L, stirring, and sealing in a container.
Furthermore, the invention also provides a method for preparing the compound chlorhexidine gargle, which comprises the following steps:
(1) weighing the raw materials and auxiliary materials according to the parts by weight;
(2) placing the weighed ethanol into a liquid preparation tank 1, sequentially adding chlorhexidine gluconate solution, metronidazole, concentrated peppermint water and green tea essence, and circularly stirring to obtain a liquid medicine A for later use;
(3) adding appropriate amount of purified water into the liquid preparation tank 2, stirring, sequentially adding glycerol, polysorbate 80, polyethylene glycol 400, and saccharin sodium, stirring for dissolving to obtain medicinal liquid B;
(4) adding the liquid medicine A into the liquid medicine B in the liquid preparation tank 2, adding the lemon yellow pigment, fixing the volume to the full volume by using purified water, circularly stirring, sampling, checking, and filling after the sampling meets relevant regulations.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention provides a method for simultaneously detecting the content of chloroaniline and chlorhexidine gluconate in compound chlorhexidine mouthrinse solution, overcomes the defect that the existing chlorhexidine gluconate detection method cannot detect the chloroaniline, and can detect the specificity, detection limit, quantitative limit, linear range, accuracy and precision of the detection method, and the result can meet the relevant requirements. The method is adopted to simultaneously detect the content of the parachloroaniline and the content of the chlorhexidine gluconate, is convenient to operate and low in cost, and can meet the requirement of actual production;
2. the compound chlorhexidine gargle prepared by the preparation method has good stability and is not easy to precipitate in the storage process.
Drawings
FIG. 1 is a chromatogram obtained according to the detection conditions of Experimental example 1;
FIG. 2 is a chromatogram obtained according to the detection conditions of Experimental example 2;
FIG. 3 is a chromatogram obtained according to the detection conditions of Experimental example 3;
FIG. 4 is a chromatogram obtained according to the detection conditions of Experimental example 4;
FIG. 5 is a chromatogram obtained according to the detection conditions of Experimental example 5;
the highest peak in the chromatogram is the peak value of chlorhexidine, and the chromatographic peak with integral is p-chloroaniline.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer and more fully describe the technical solutions in the embodiments of the present invention, it is obvious that the described embodiments are a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
EXAMPLE 1 preparation of Compound chlorhexidine mouthrinse
The method comprises the following steps:
(1) preparation of concentrated peppermint water
Adding purified water into 12L 90% ethanol, stirring, slowly adding peppermint oil 0.4L, stirring, adding purified water to constant volume of 20L, stirring, and sealing in container to obtain concentrated peppermint water.
(2) Weighing the following raw and auxiliary materials in parts by weight:
2400 parts of chlorhexidine gluconate solution, 400 parts of metronidazole, 100 parts of glycerol, 802 parts of polysorbate, 100 parts of polyethylene glycol 400, 20 parts of concentrated peppermint water, 110 parts of ethanol, 200 parts of sodium saccharinate, 6 parts of lemon yellow, 2 parts of green tea essence and 1650 parts of purified water;
(3) placing the weighed ethanol into a liquid preparation tank 1, sequentially adding chlorhexidine gluconate solution, metronidazole, concentrated peppermint water and green tea essence, and circularly stirring to obtain a liquid medicine A for later use;
(4) adding appropriate amount of purified water into the liquid preparation tank 2, stirring, sequentially adding glycerol, polysorbate 80, polyethylene glycol 400, and saccharin sodium, stirring for dissolving to obtain medicinal liquid B;
(5) adding the liquid medicine A into the liquid medicine B in the liquid preparation tank 2, adding the lemon yellow pigment, fixing the volume to the full volume by using purified water, circularly stirring, sampling, checking, and filling after the sampling meets relevant regulations.
EXAMPLE 2 preparation of Compound chlorhexidine mouthrinse
The method comprises the following steps:
(1) preparation of concentrated peppermint water
Adding purified water into 12L 90% ethanol, stirring, slowly adding peppermint oil 0.4L, stirring, adding purified water to constant volume of 20L, stirring, and sealing in container to obtain concentrated peppermint water.
(2) Weighing the following raw and auxiliary materials in parts by weight:
3000 parts of chlorhexidine gluconate solution, 350 parts of metronidazole, 120 parts of glycerol, 803 parts of polysorbate, 80 parts of polyethylene glycol 400, 25 parts of concentrated peppermint water, 100 parts of ethanol, 250 parts of sodium saccharinate, 3 parts of lemon yellow, 3 parts of green tea essence and 1650 parts of purified water.
(3) Placing the weighed ethanol into a liquid preparation tank 1, sequentially adding chlorhexidine gluconate solution, metronidazole, concentrated peppermint water and green tea essence, and circularly stirring to obtain a liquid medicine A for later use;
(4) adding into purified water solution tank 2, stirring, sequentially adding glycerol, polysorbate 80, polyethylene glycol 400, and saccharin sodium, stirring for dissolving to obtain medicinal liquid B;
(5) adding the liquid medicine A into the liquid medicine B in the liquid preparation tank 2, adding the lemon yellow pigment, fixing the volume to the full volume by using purified water, circularly stirring, sampling, checking, and filling after the sampling meets relevant regulations.
EXAMPLE 3 preparation of Compound chlorhexidine mouthrinse
The method comprises the following steps:
(1) preparation of concentrated peppermint water
Adding purified water into 12L 90% ethanol, stirring, slowly adding peppermint oil 0.4L, stirring, adding purified water to constant volume of 20L, stirring, and sealing in container to obtain concentrated peppermint water.
(2) Weighing the following raw and auxiliary materials in parts by weight:
2000 parts of chlorhexidine gluconate solution, 450 parts of metronidazole, 50 parts of glycerol, 801 parts of polysorbate, 40050 parts of polyethylene glycol, 10 parts of concentrated peppermint water, 150 parts of ethanol, 300 parts of sodium saccharinate, 5 parts of lemon yellow, 4 parts of green tea essence and 1650 parts of purified water
(3) Placing the weighed ethanol into a liquid preparation tank 1, sequentially adding chlorhexidine gluconate solution, metronidazole, concentrated peppermint water and green tea essence, and circularly stirring to obtain a liquid medicine A for later use;
(4) adding into purified water solution tank 2, stirring, sequentially adding glycerol, polysorbate 80, polyethylene glycol 400, and saccharin sodium, stirring for dissolving to obtain medicinal liquid B;
(5) adding the liquid medicine A into the liquid medicine B in the liquid preparation tank 2, adding the lemon yellow pigment, fixing the volume to the full volume by using purified water, circularly stirring, sampling, checking, and filling after the sampling meets relevant regulations.
Example 4 establishment of a method for detecting the content of chloraniline and chlorhexidine gluconate in a Compound chlorhexidine gargle
As the parachloroaniline cannot adopt a method for detecting related substances or content of the chlorhexidine gluconate in the existing batch quality standard, the methodology development is carried out again. Meanwhile, in order to save the inspection cost, the chlorhexidine gluconate detection method is researched again, and the same method is adopted to detect the content of the parachloroaniline and the chlorhexidine gluconate.
1. Method for detecting content of parachloroaniline and chlorhexidine gluconate
Experimental example 1
Chromatographic conditions are as follows: a sulfonic bonded silica gel chromatographic column is used as a filling agent, the detection wavelength is 239nm, the column temperature is 40 ℃, and the sample injection amount is 50 mu l.
Preparation of a mobile phase: 27.6g of sodium dihydrogen phosphate, 10ml of triethylamine and 2000ml of water were weighed out and the pH was adjusted to 3.0 with phosphoric acid as mobile phase A. Acetonitrile is mobile phase B.
Preparing a test solution: p-chloroaniline (12 mg) was weighed precisely and placed in a 100ml measuring flask as a p-chloroaniline stock solution. Precisely measuring 5ml of compound chlorhexidine gargle and 0.8ml of parachloroaniline stock solution, placing into a 100ml measuring flask, diluting with a diluting solvent to scale, and shaking up to obtain the compound chlorhexidine gargle.
Gradient elution was performed according to table 1 below and the chromatogram is shown in figure 1.
TABLE 1
Figure BDA0002610865620000071
Experimental example two
The chromatographic conditions, the preparation of mobile phase and the preparation of test solution are the same as those in the first experimental example, gradient elution is carried out according to the following table 2, and the chromatogram is shown in fig. 2.
TABLE 2
Figure BDA0002610865620000072
Experimental example III
The chromatographic conditions, the preparation of mobile phase and the preparation of test solution are the same as those in the first experimental example, gradient elution is carried out according to the following table 3, and the chromatogram is shown in fig. 3.
TABLE 3
Figure BDA0002610865620000073
Figure BDA0002610865620000081
Experimental example four
The chromatographic conditions and the preparation of the test solution are the same as those of the first experimental example, and the preparation of the mobile phase is as follows: weighing 27.6g of sodium dihydrogen phosphate, adding 10ml of triethylamine, adding 2000ml of water, adjusting the pH to 3.0 by using phosphoric acid, taking 700ml of the phosphate solution, adding 300ml of acetonitrile, and mixing to obtain a mobile phase A; acetonitrile is mobile phase B. Gradient elution was performed according to Table 4 below and the chromatogram is shown in FIG. 4.
TABLE 4
Figure BDA0002610865620000082
Experimental example five
The chromatographic conditions, the preparation of mobile phase and the determination method are the same as those of the first experimental example, gradient elution is carried out according to the following table 5, and the chromatogram is shown in fig. 5.
TABLE 5
Figure 1
And (4) conclusion: the test results of the experimental examples I to V can show that the detection method of the experimental example 5 can better adjust the peak emergence time of the parachloroaniline and the chlorhexidine, so that the parachloroaniline and the chlorhexidine are basically separated from the adjacent auxiliary material peak or solvent peak and are not interfered by the auxiliary material peak or the solvent peak. Therefore, the detection method of Experimental example 5 was used.
2. Establishment of method for detecting content of parachloroaniline and chlorhexidine gluconate
In conclusion, the method for detecting the content of the parachloroaniline and the chlorhexidine gluconate is formulated as follows:
(1) preparation of test solution
Precisely measuring 5ml of compound chlorhexidine gargle to be measured, placing the gargle in a 100ml measuring flask, diluting the gargle to a scale with a diluent solvent, and shaking up to be used as a test solution; wherein the diluting solvent is prepared by the following method: 27.6g of sodium dihydrogen phosphate is weighed, 2000ml of water is added, and the pH is adjusted to 3.0 by phosphoric acid;
(2) preparation of reference stock solution
Accurately weighing 12mg of chlorhexidine acetate, placing the chlorhexidine acetate in a 50ml measuring flask, adding 5ml of acetonitrile and a proper amount of diluting solvent, dissolving by ultrasonic, diluting to scale with the diluting solvent, shaking up, and using the chlorhexidine acetate as a reference stock solution; accurately weighing 12mg of parachloroaniline, placing the parachloroaniline into a 100ml measuring flask, adding a proper amount of a diluting solvent, performing ultrasonic dissolution, diluting the parachloroaniline to a scale with the diluting solvent, and shaking up to be used as a parachloroaniline reference substance stock solution;
(3) preparation of Mixed control solutions
Precisely measuring the reference stock solution obtained in the step (2), diluting the reference stock solution into a mixed reference stock solution containing 42 mu g of chlorhexidine acetate and 0.12 mu g of parachloroaniline in each 1ml of diluted solvent, and shaking up to obtain a reference stock solution;
(4) liquid chromatograph detection
Precisely measuring the sample solution and the control solution by 50 μ l each, respectively injecting into a liquid chromatograph, using a sulfonic bonded silica gel chromatographic column as a filler, using a phosphate solution (weighing 27.6g of sodium dihydrogen phosphate, adding 10ml of triethylamine, adding 2000ml of water, adjusting pH to 3.0 with phosphoric acid) as a mobile phase A, and using acetonitrile as a mobile phase B; gradient elution was performed as in table 5; the detection wavelength is 239nm, and the column temperature is 40 ℃. Precisely measuring 50 μ l of the reference solution, injecting into chromatograph, and recording chromatogram. If a chromatographic peak with the same retention time as parachloroaniline in the chromatogram of the reference solution exists in the chromatogram of the test solution, calculating according to the peak area by an external standard method; the chlorhexidine gluconate is calculated by peak area according to an external standard method, and the result is multiplied by 1.4352 to obtain the chlorhexidine gluconate;
the calculation formula calculated by peak area according to an external standard method is as follows:
Figure BDA0002610865620000091
Figure BDA0002610865620000092
Cfor supplying to-the concentration of the component to be tested in the test sample; cTo pair-the concentration of the component to be measured in the control;
Afor supplying toPeak area of the component to be measured in the test sample; a. theTo pairPeak area of the component to be measured in the control;
Ssign boardThe marked amount of the product.
Limitation: the content of chlorhexidine containing glucose is 90.0-110.0% of the marked amount; the content of p-chloroaniline must not exceed 0.2% of the limit.
3. Methodology validation
The invention sets out a detection method for the content of the parachloroaniline and the chlorhexidine gluconate by comparing experimental examples 1-5, and verifies the specificity, detection limit, quantitative limit, linear range, accuracy, precision and sample solution placement stability of the finally established detection method, and the result is shown in the following tables 6 and 7.
TABLE 6 Table of the results of the methodological examination of p-chloroaniline
Figure BDA0002610865620000101
Figure BDA0002610865620000111
Figure BDA0002610865620000121
TABLE 7 tables of the results of the methodological verification of chlorhexidine content
Figure BDA0002610865620000122
Figure BDA0002610865620000131
Figure BDA0002610865620000141
4. Accuracy test
The range is as follows: 3 different concentrations (namely 80%, 100% and 120% sample concentrations) are designed, and 3 parts of test solution is prepared for each concentration and is measured according to a content measurement method.
Preparing blank auxiliary materials: preparing blank auxiliary materials according to requirements.
Preparing a reference substance solution: accurately weighing appropriate amount of chlorhexidine acetate and p-chloroaniline, diluting with diluent solvent to obtain mixed control solution containing chlorhexidine acetate 42 μ g and p-chloroaniline 0.1 μ g per 1ml, and shaking up to obtain the control solution.
Preparing a test solution: taking compound chlorhexidine gargle (prepared in example 1), precisely weighing 5ml, placing into a 100ml measuring flask, diluting to scale with diluent solvent (weighing 27.6g sodium dihydrogen phosphate, adding 2000ml water, adjusting pH to 3.0 with phosphoric acid), and shaking uniformly to obtain sample solution.
Preparation of a reference stock solution:
solution a: accurately weighing 12mg of parachloroaniline, placing the parachloroaniline in a 100ml measuring flask, diluting the parachloroaniline to a scale by using a diluting solvent, accurately transferring 2ml to 100ml measuring flasks, diluting the parachloroaniline to the scale by using the diluting solvent, and shaking up the flasks to obtain the parachloroaniline liquid.
Solution b: accurately weighing 44mg of chlorhexidine acetate, placing the chlorhexidine acetate in a 50ml measuring flask, adding 5ml of acetonitrile, diluting to scale with a diluting solvent, and shaking up to obtain the chlorhexidine acetate.
80% test solution: precisely measuring 5ml of blank auxiliary materials and 4ml → 100ml of solutions a and b respectively;
100% test solution: precisely measuring 5ml of blank auxiliary materials and 5ml → 100ml of solutions a and b respectively;
120% test solution: precisely measuring 5ml of blank auxiliary materials and 6ml → 100ml of solutions a and b respectively;
preparing 3 parts of test solution for each concentration, respectively, preparing 9 parts of test solution for each concentration, respectively diluting the test solutions to a scale with a diluent, and shaking up to obtain the test solution.
Precisely measuring the reference solution and each sample solution by 50 μ l, respectively injecting into a liquid chromatograph, recording chromatogram, and calculating the recovery rate, average recovery rate and relative standard deviation of parachloroaniline and chlorhexidine in the sample by peak area according to external standard method. The results are shown in tables 8 and 9.
TABLE 8 measurement accuracy test results for p-chloroaniline content
Figure BDA0002610865620000151
TABLE 9 Chlorhexidine gluconate content determination accuracy test results
Figure BDA0002610865620000152
Calculating the formula:
Figure BDA0002610865620000161
as a result: the average recovery rate of the accuracy (recovery rate) of the method for measuring the content of the parachloroaniline is 104.26 percent, the recovery rate result is in the range of 90 to 110 percent, and the relative deviation of the recovery rates of 9 test sample solutions is 1.81 percent and is less than 2.0 percent. The average recovery rate of the glucose chlorhexidine content measuring method is 98.72 percent, the recovery rate result is in the range of 98 percent to 101 percent, and the relative deviation of the recovery rate of 9 test sample solutions is 0.92 percent. Meets the requirements.
And (4) conclusion: the content determination method has good recovery rate and is accurate and feasible.
5. Stability test of solution
The determination method comprises the following steps:
preparing a reference substance solution: accurately weighing appropriate amount of chlorhexidine acetate and p-chloroaniline, diluting with a diluent solvent to obtain a mixed reference solution containing 42 μ g of chlorhexidine acetate and 0.12 μ g of p-chloroaniline per 1ml, and shaking up to obtain the reference solution.
Preparing a test solution: taking compound chlorhexidine gargle (prepared in example 1), precisely weighing 5ml, placing into a 100ml measuring flask, diluting to scale with diluent solvent (weighing 27.6g sodium dihydrogen phosphate, adding 2000ml water, adjusting pH to 3.0 with phosphoric acid), and shaking uniformly to obtain sample solution. The results are shown in tables 10 and 11.
TABLE 10 Table of the results of the test for the stability of the control solutions
Figure BDA0002610865620000162
Figure BDA0002610865620000171
TABLE 11 table of the test results of the stability of the test solutions
Figure BDA0002610865620000172
As a result: RSD of the p-chloroaniline peak area in the reference solution is less than 10%, and RSD of the glucose chlorhexidine peak area is less than 2%; the difference between the parachloroaniline content in the test solution and the time 0 is within +/-10%, and the difference between the chlorhexidine gluconate content and the time 0 is within +/-2%.
And (4) conclusion: the control solution and the test solution have good stability within 18 hours.
To summarize: the invention relates to a preparation method and a quality detection method of a compound chlorhexidine gargle, which develops the same method for detecting the content of parachloroaniline and chlorhexidine gluconate by researching the content detection method of the related substances of the compound chlorhexidine gargle, and saves the inspection cost while controlling the product quality.

Claims (8)

1. A method for detecting the content of chloroaniline and chlorhexidine gluconate in a compound chlorhexidine gargle is characterized by comprising the following steps:
(1) preparation of test solution
Precisely measuring 5ml of compound chlorhexidine gargle to be measured, placing the gargle in a 100ml measuring flask, diluting the gargle to a scale with a diluent solvent, and shaking up to be used as a test solution; wherein the diluting solvent is prepared by the following method: 27.6g of sodium dihydrogen phosphate is weighed, 2000ml of water is added, and the pH is adjusted to 3.0 by phosphoric acid;
(2) preparation of reference stock solution
Accurately weighing 12mg of chlorhexidine acetate, placing the chlorhexidine acetate in a 50ml measuring flask, adding 5ml of acetonitrile and a proper amount of diluting solvent, dissolving by ultrasonic, diluting to scale with the diluting solvent, shaking up, and using the chlorhexidine acetate as a reference stock solution; accurately weighing 12mg of parachloroaniline, placing the parachloroaniline into a 100ml measuring flask, adding a proper amount of a diluting solvent, performing ultrasonic dissolution, diluting the parachloroaniline to a scale with the diluting solvent, and shaking up to be used as a parachloroaniline reference substance stock solution;
(3) preparation of Mixed control solutions
Precisely measuring the reference stock solution obtained in the step (2), diluting the reference stock solution into a mixed reference stock solution containing 42 mu g of chlorhexidine acetate and 0.12 mu g of parachloroaniline in each 1ml of diluted solvent, and shaking up to obtain a reference stock solution;
(4) liquid chromatograph detection
Precisely measuring the sample solution and the reference solution by 50 μ l each, injecting into a liquid chromatograph, and recording chromatogram; if a chromatographic peak with the retention time consistent with that of parachloroaniline in a chromatogram of a reference solution exists in the chromatogram of the test solution, calculating the content of the parachloroaniline according to an external standard method by using a peak area; the chlorhexidine gluconate is calculated by peak area according to an external standard method, and the result is multiplied by 1.4352 to obtain the chlorhexidine gluconate;
the calculation formula calculated by peak area according to an external standard method is as follows:
Figure FDA0002610865610000011
Figure FDA0002610865610000012
Cfor supplying to-the concentration of the component to be tested in the test sample; cTo pair-the concentration of the component to be measured in the control;
Afor supplying toPeak area of the component to be measured in the test sample; a. theTo pairPeak area of the component to be measured in the control;
Ssign boardThe marked amount of the product.
2. The method of claim 1, wherein the detection conditions of the liquid chromatography in step (4) are: a sulfonic bonded silica gel chromatographic column is used as a filling agent, a phosphate solution is used as a mobile phase A, and acetonitrile is used as a mobile phase B; gradient elution was performed in the following order:
Figure FDA0002610865610000021
the detection wavelength is 239nm, and the column temperature is 40 ℃.
3. The method of claim 2, wherein the phosphate solution is prepared by: 27.6g of sodium dihydrogen phosphate was weighed, 10ml of triethylamine was added, 2000ml of water was added, and pH was adjusted to 3.0 with phosphoric acid.
4. The method of claim 1, wherein the chlorhexidine-containing glucose is present in an amount of from 90.0% to 110.0% of the labeled amount; the content of p-chloroaniline must not exceed 0.2% of the limit.
5. The compound chlorhexidine gargle is characterized by being prepared from the following raw and auxiliary materials in parts by weight: 3000 parts of chlorhexidine gluconate solution, 500 parts of metronidazole 300, 50-150 parts of glycerol, 801-3 parts of polysorbate, 150 parts of polyethylene glycol 40050, 10-30 parts of concentrated peppermint water, 50-150 parts of ethanol, 300 parts of sodium saccharinate essence, 2-10 parts of lemon yellow, 1-5 parts of green tea essence and 2000 parts of purified water.
6. The compound chlorhexidine gargle of claim 5, which is composed of the following raw and auxiliary materials in parts by weight: 2400 parts of chlorhexidine gluconate solution, 400 parts of metronidazole, 100 parts of glycerol, 802 parts of polysorbate, 100 parts of polyethylene glycol 400, 20 parts of concentrated peppermint water, 110 parts of ethanol, 200 parts of sodium saccharinate, 6 parts of lemon yellow, 2 parts of green tea essence and 1650 parts of purified water.
7. The compound chlorhexidine gargle of claim 5 or 6, wherein the concentrated peppermint water is prepared by the following method:
adding purified water into 12L of 90% ethanol, stirring, slowly adding 0.4L of dementholized peppermint oil, stirring, adding purified water to constant volume of 20L, stirring, and sealing in a container.
8. A method for preparing the compound chlorhexidine gargle of any of claims 5-7, comprising the steps of:
(1) weighing the raw and auxiliary materials according to the parts by weight of the raw and auxiliary materials in the claim 4 or 5;
(2) placing the weighed ethanol into a liquid preparation tank 1, sequentially adding chlorhexidine gluconate solution, metronidazole, concentrated peppermint water and green tea essence, and circularly stirring to obtain a liquid medicine A for later use;
(3) adding appropriate amount of purified water into the liquid preparation tank 2, stirring, sequentially adding glycerol, polysorbate 80, polyethylene glycol 400, and saccharin sodium, stirring for dissolving to obtain medicinal liquid B;
(4) adding the liquid medicine A into the liquid medicine B in the liquid preparation tank 2, adding the lemon yellow pigment, fixing the volume to the full volume by using purified water, circularly stirring, sampling, checking, and filling after the sampling meets relevant regulations.
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