阅读说明：本技术 复方香波中硝酸咪康唑和葡萄糖酸氯己定的含量检测方法 (Method for detecting content of miconazole nitrate and chlorhexidine gluconate in compound shampoo ) 是由 徐奇清 孙迎 聂姣 林孝崇 赵奉君 于 2018-07-23 设计创作，主要内容包括：本发明涉及一种复方香波中硝酸米康唑和葡萄糖酸氯己定的含量检测方法,其采用高效液相色谱外标法同时测定复方香波中硝酸咪康唑和葡萄糖酸氯己定的含量。该检测方法的条件响应值高,在保证待测样品中组分分离度的情况下出峰快,硝酸咪康唑的保留时间约为3.0min左右、葡萄糖酸氯己定的保留时间为3.9min左右,能够快速测定被测组分的含量,大大提高了检测工作效率和检测结果的准确性,有利于控制产品的质量。(The invention relates to a method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo, which adopts a high performance liquid chromatography external standard method to simultaneously determine the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo. The detection method has high condition response value, is quick in peak emergence under the condition of ensuring the separation degree of components in a sample to be detected, can quickly determine the content of the detected components, greatly improves the detection working efficiency and the accuracy of a detection result, and is beneficial to controlling the quality of products, and the retention time of miconazole nitrate is about 3.0min and the retention time of chlorhexidine gluconate is about 3.9 min.)

1. A method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo is characterized in that the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo is simultaneously determined by adopting high performance liquid chromatography, and comprises the following steps:

diluting a sample to be detected by using a solvent to obtain a test solution; the dosage ratio of the sample to be detected to the solvent is 1 g: (100-200) mL;

injecting 10-20 mu L of the sample solution into a high performance liquid chromatograph for detection to obtain the content of miconazole nitrate and chlorhexidine gluconate in the sample to be detected;

the chromatographic conditions in the high performance liquid chromatography detection are as follows:

a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent;

mobile phase: the catalyst consists of acetonitrile, phosphate solution and triethylamine according to the volume ratio of (35-50): 50-65): 1, and the pH value is 3 +/-0.1; the concentration of the phosphate solution is 0.02 mol/L-0.08 mol/L;

flow rate: 0.5mL/min to 1.5 mL/min;

column temperature: 25-35 ℃;

detection wavelength: 210nm to 240 nm;

the solvent is of the same composition as the mobile phase.

2. The method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo as claimed in claim 1, wherein the mobile phase consists of acetonitrile, phosphate solution and triethylamine in a volume ratio of (40-50): (50-60): 1.

3. The method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo as claimed in claim 2, wherein the mobile phase consists of acetonitrile, phosphate solution and triethylamine in a volume ratio of 45:55: 1.

4. The compound shampoo as claimed in claim 1, wherein the phosphate solution is disodium hydrogen phosphate solution.

5. The method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo as claimed in claim 1, wherein the detection wavelength is 230 nm.

6. The method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo as claimed in claim 1, wherein the chromatographic column is ZORBZX Eclipse XDB-C18Analytical 4.6 x 150mm 5-Micron or Kromasil 100-5-C184.6 x 150 mm.

7. The method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo as claimed in claim 1, wherein the flow rate is 0.8mL/min to 1.2 mL/min.

8. The method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo as claimed in claim 1, wherein the content of chlorhexidine gluconate in the sample solution is 0.02mg/mL to 0.2mg/mL, and the content of miconazole nitrate is 0.02mg/mL to 0.2 mg/mL.

9. The method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo as claimed in any one of claims 1 to 8, wherein the regulator used for regulating the pH value of the mobile phase is phosphoric acid.

10. The method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo as claimed in any one of claims 1 to 8, characterized by further comprising the following steps:

performing HPLC detection by using a miconazole nitrate reference substance according to an external standard method to obtain the relation between the content of miconazole nitrate and the peak area, and calculating to obtain the content of miconazole nitrate in the sample to be detected according to the relation between the content of miconazole nitrate and the peak area of miconazole nitrate in a chromatogram obtained by detecting the sample solution in a high performance liquid chromatograph;

and performing HPLC detection on a chlorhexidine gluconate reference substance according to an external standard method to obtain the relation between the chlorhexidine gluconate and the peak area, and calculating to obtain the content of the chlorhexidine gluconate in the sample to be detected according to the relation between the content of the chlorhexidine gluconate and the peak area of the chlorhexidine gluconate in a chromatogram obtained by detecting the sample solution in a high performance liquid chromatograph.

Technical Field

The invention relates to the technical field of veterinary drug detection, in particular to a method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo.

Background

The shampoo is a transliteration of the foreign language 'shampoo', mainly comprises a detergent, an auxiliary detergent and an additive, and mainly has the effects of cleaning and caring hair; the additive can meet the special requirements of the shampoo and endow the shampoo with various different efficacies, so that the effect of the shampoo is continuously refined, and the compound shampoo preparation prepared by adding the miconazole nitrate and the chlorhexidine gluconate with antibacterial effect is a novel veterinary medicine shampoo preparation for pets.

The content detection method of the miconazole nitrate adopts a potentiometric titration method to determine, and the titration solution is 0.1mol/L perchloric acid titration solution; the potentiometric titration method is a method for determining a titration end point by measuring potential change in a titration process, has certain limitations in application, the sensitivity of the method may influence the accuracy of a detection result when detecting a low-concentration drug, perchloric acid is an inorganic acid with strong acidity, has strong oxidizing property, is easily decomposed at room temperature, has unstable factors as a titration solution, and the change of concentration easily causes errors in the detection result. The chlorhexidine gluconate is detected by adopting an ultraviolet-visible spectrophotometry method, and the detection method has the defect of relatively low accuracy. In the chinese pharmacopoeia, High Performance Liquid Chromatography (HPLC) is adopted to control only related substances in miconazole nitrate raw materials or single preparations thereof, and a detection method of a compound preparation thereof is not described. The chlorhexidine gluconate content in the chlorhexidine gluconate gargle is detected by adopting a gradient elution high performance liquid chromatography, but the elution peak time of the method is slow, and the response value is low. And the results of experiments prove that the traditional detection method and the method recorded in the Chinese pharmacopoeia can not be applied to the content determination of the miconazole nitrate and the chlorhexidine gluconate in the compound shampoo; at present, no quality standard and effective method for detecting the content of miconazole nitrate and chlorhexidine gluconate exist in the preparation.

Disclosure of Invention

Based on the content detection method, the content detection method of the miconazole nitrate and the chlorhexidine gluconate in the compound shampoo can be used for simultaneously, quickly and effectively detecting the content of the miconazole nitrate and the content of the chlorhexidine gluconate.

A method for detecting the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo adopts high performance liquid chromatography to simultaneously determine the content of miconazole nitrate and chlorhexidine gluconate in compound shampoo, and comprises the following steps:

diluting a sample to be detected by using a solvent to obtain a test solution; the dosage ratio of the sample to be detected to the solvent is 1 g: 100-200) mL;

injecting 10-20 mu L of the sample solution into a high performance liquid chromatograph for detection to obtain the content of miconazole nitrate and chlorhexidine gluconate in the sample to be detected;

the chromatographic conditions in the high performance liquid chromatography detection are as follows:

a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent;

mobile phase: the catalyst consists of acetonitrile, phosphate solution and triethylamine according to the volume ratio of (35-50): 50-65): 1, and the pH value is 3 +/-0.1; the concentration of the phosphate solution is 0.02 mol/L-0.08 mol/L;

flow rate: 0.5mL/min to 1.5 mL/min;

column temperature: 25-35 ℃;

detection wavelength: 210nm to 240 nm;

the solvent is of the same composition as the mobile phase.

In one embodiment, the mobile phase consists of acetonitrile, phosphate solution and triethylamine according to the volume ratio of (40-50): (50-60): 1.

Further, the mobile phase consists of acetonitrile, phosphate solution and triethylamine in a volume ratio of 45:55: 1.

In one embodiment, the detection wavelength is 230 nm.

In one embodiment, the phosphate solution is a disodium hydrogen phosphate solution.

In one embodiment, the column is ZORBZX Eclipse XDB-C18Analytical 4.6X 150mm 5-Micron or Kromasil 100-5-C184.6X 150 mm.

Preferably, the column is ZORBZX Eclipse XDB-C18Analytical 4.6X 150mm 5-Micron.

In one embodiment, the flow rate is 0.8mL/min to 1.2 mL/min.

In one embodiment, the sample solution contains chlorhexidine gluconate 0.02 mg/mL-0.2 mg/mL and miconazole nitrate 0.02 mg/mL-0.2 mg/mL.

In one embodiment, the adjusting agent used to adjust the pH of the mobile phase is phosphoric acid.

In one embodiment, the detection method further comprises the following steps:

performing HPLC detection by using a miconazole nitrate reference substance according to an external standard method to obtain the relation between the content of miconazole nitrate and the peak area, and calculating to obtain the content of miconazole nitrate in the sample to be detected according to the relation between the content of miconazole nitrate and the peak area of miconazole nitrate in a chromatogram obtained by detecting the sample solution in a high performance liquid chromatograph;

and performing HPLC detection on a chlorhexidine gluconate reference substance according to an external standard method to obtain the relation between the chlorhexidine gluconate and the peak area, and calculating to obtain the content of the chlorhexidine gluconate in the sample to be detected according to the relation between the content of the chlorhexidine gluconate and the peak area of the chlorhexidine gluconate in a chromatogram obtained by detecting the sample solution in a high performance liquid chromatograph.

Specifically, a proper amount of miconazole nitrate reference substance is taken and prepared into a miconazole nitrate reference sample containing 0.02 mg/mL-0.2 mg/mL in the solvent, sample treatment and detection are carried out in parallel with the test solution, a chromatogram is recorded, the concentration of miconazole nitrate corresponding to the peak area in the chromatogram is measured, and a standard curve is drawn;

and measuring the peak area of the miconazole nitrate in the chromatogram obtained by injecting the test solution into a high performance liquid chromatograph for detection, and calculating to obtain the content of the miconazole nitrate in the sample to be detected according to the standard curve.

The detection method adopts a high-efficiency liquid phase external standard method to simultaneously determine the content of miconazole nitrate and chlorhexidine gluconate in the compound shampoo preparation, selects a proper mobile phase component, adjusts the proportion composition of the mobile phase and combines other parameter conditions, has high response value of detection conditions, can well separate the detected component in a sample to be detected, has quick peak emergence of miconazole nitrate and chlorhexidine gluconate, can effectively shorten the retention time of the detected component, quickly and accurately determines the content of miconazole nitrate and chlorhexidine gluconate simultaneously, solves the problem of determining the content of miconazole nitrate and chlorhexidine gluconate in the compound shampoo, greatly improves the detection working efficiency, and provides an effective detection method for better controlling the product quality.

Drawings

FIG. 1 is an HPLC detection chromatogram of compound miconazole nitrate shampoo of example 1;

FIG. 2 is an HPLC detection chromatogram of a miconazole nitrate control of example 1;

FIG. 3 is the HPLC detection chromatogram of the chlorhexidine gluconate control of example 1.

Detailed Description

In order that the invention may be more fully understood, a more particular description of the invention will now be rendered by reference to specific embodiments thereof that are illustrated in the appended drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The compound miconazole nitrate shampoo is self-made by Australian dragons pharmaceutical Co., Ltd, south China sea, in Foshan City, and mainly comprises miconazole nitrate, chlorhexidine gluconate, a detergent and various additives; miconazole nitrate controls and chlorhexidine gluconate controls were purchased from Siga corporation; acetonitrile, triethylamine and phosphoric acid are used as chromatographic purity, and disodium hydrogen phosphate is used as analytical purity.