阅读说明：本技术 葡萄糖酸氯己定的含量检测方法 (Content detection method of chlorhexidine gluconate ) 是由 徐奇清 孙迎 聂姣 付良凯 赵奉君 于 2018-07-23 设计创作，主要内容包括：本发明涉及一种葡萄糖酸氯己定的含量检测方法,用溶剂将待测样品稀释成葡萄糖酸氯己定的含量为0.02mg/mL～0.2mg/mL的供试品溶液；取10μL～20μL所述供试品溶液注入高效液相色谱仪中进行检测,得到所述待测样品中葡萄糖酸氯己定的含量；所述高效液相色谱检测时的色谱条件为：色谱柱：以十八烷基硅烷键合硅胶为填充剂；流动相：乙腈、磷酸盐溶液和三乙胺的体积比为(35～50):(50～65):1,pH值为3±0.1,所述磷酸盐溶液的浓度为0.02mol/L～0.08mol/L；流速：0.5mL/min～1.5mL/min；柱温：25℃～35℃；检测波长：258～259nm；所述溶剂与所述流动相相同。该方法条件响应值高,在保证组分分离度的情况下,葡萄糖酸氯己定出峰快,能够实现快速测定其含量,提高了检测工作效率,有利于控制产品的质量。(The invention relates to a method for detecting the content of chlorhexidine gluconate, which comprises the steps of diluting a sample to be detected into a sample solution with the content of chlorhexidine gluconate of 0.02 mg/mL-0.2 mg/mL by using a solvent; injecting 10-20 mu L of the sample solution into a high performance liquid chromatograph for detection to obtain the content of 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 volume ratio of acetonitrile to phosphate solution to triethylamine is (35-50): (50-65): 1, the pH value is 3 +/-0.1, and the concentration of the phosphate solution is 0.02-0.08 mol/L; flow rate: 0.5mL/min to 1.5 mL/min; column temperature: 25-35 ℃; detection wavelength: 258 to 259 nm; the solvent is the same as the mobile phase. The method has high condition response value, ensures the component separation degree, has fast peak emergence of the chlorhexidine gluconate, can realize the fast determination of the content of the chlorhexidine gluconate, improves the detection working efficiency and is beneficial to controlling the quality of products.)

1. A method for detecting the content of chlorhexidine gluconate is characterized by comprising the following steps:

diluting a sample to be detected into a sample solution to be detected with a solvent, wherein the content of chlorhexidine gluconate is 0.02 mg/mL-0.2 mg/mL;

injecting 10-20 mu L of the sample solution into a high performance liquid chromatograph for detection to obtain the content of 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 volume ratio of acetonitrile to phosphate solution to triethylamine is (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: 258nm to 259 nm;

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

2. The method as claimed in claim 1, wherein the sample is a shampoo containing chlorhexidine gluconate.

3. The method for detecting the content of chlorhexidine gluconate according to claim 1, wherein the volume ratio of the acetonitrile, the phosphate solution and the triethylamine in the mobile phase is (40-50): (50-60): 1.

4. The method for detecting the content of chlorhexidine gluconate according to claim 3, wherein the volume ratio of the acetonitrile, the phosphate solution and the triethylamine in the mobile phase is 45:55: 1.

5. The method for detecting the content of chlorhexidine gluconate according to claim 1, wherein the phosphate solution is a disodium hydrogen phosphate solution.

6. The method for detecting the content of chlorhexidine gluconate according to claim 1, wherein the chromatographic column is ZORBZX Eclipse XDB-C18Analytical4.6 x 150mm 5-Micron or Kromasil 100-5-C184.6 x 150 mm.

7. The method for detecting the content of chlorhexidine gluconate according to claim 6, wherein the flow rate is 0.8mL/min to 1.2 mL/min.

8. The method as claimed in claim 1, wherein the chlorhexidine gluconate solution has a chlorhexidine gluconate content of 0.05mg/mL to 0.15 mg/mL.

9. The method for detecting the content of chlorhexidine gluconate according to any of claims 1 to 8, wherein the regulator used for regulating the pH of the mobile phase is phosphoric acid.

10. The method for detecting the content of chlorhexidine gluconate according to any one of claims 1 to 8, further comprising the steps of:

and carrying out HPLC detection by using a chlorhexidine gluconate reference substance according to an external standard method to obtain the relation between the content of the chlorhexidine gluconate and the peak area, and calculating 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 chemical detection, in particular to a content detection method of chlorhexidine gluconate.

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 effects, so that the effect of the shampoo is continuously refined, and the compound shampoo preparation prepared by adding medicinal components with antibacterial effect such as chlorhexidine gluconate and the like is a novel veterinary medicine shampoo preparation for pets.

At present, no quality standard aiming at the preparation and no effective method for detecting the content of the chlorhexidine gluconate exist. The content detection method of dilute chlorhexidine gluconate solution in Chinese pharmacopoeia adopts ultraviolet-visible spectrophotometry for detection, and the detection method has low accuracy; the content detection of the chlorhexidine gluconate in the chlorhexidine gluconate gargle is performed by adopting High Performance Liquid Chromatography (HPLC) of gradient elution, but the method has the advantages of slow elution peak time, 21 minutes of elution time, long time consumption and low response value; the experimental verification that the method is adopted to measure the content of the chlorhexidine gluconate in the shampoo shows that the method is not suitable for measuring the content of the chlorhexidine gluconate in the shampoo preparation.

Disclosure of Invention

Therefore, the method for detecting the content of the chlorhexidine gluconate can be used for quickly and effectively detecting the content of the chlorhexidine gluconate in shampoo containing the chlorhexidine gluconate.

A method for detecting the content of chlorhexidine gluconate comprises the following steps:

diluting a sample to be detected into a sample solution to be detected with a solvent, wherein the content of chlorhexidine gluconate is 0.02 mg/mL-0.2 mg/mL;

injecting 10-20 mu L of the sample solution into a high performance liquid chromatograph for detection to obtain the content of 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 volume ratio of acetonitrile to phosphate solution to triethylamine is (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: 258nm to 259 nm;

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

The content of chlorhexidine gluconate is not limited to mg/mL, but may be other concentration units such as mass percentage content and molarity.

Preferably, the detection wavelength is 259 nm.

In one embodiment, the sample to be tested is a shampoo containing chlorhexidine gluconate.

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

In one embodiment, the volume ratio of acetonitrile, phosphate solution and triethylamine in the mobile phase is 45:55: 1.

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

Further, the concentration of the disodium hydrogen phosphate solution is 0.05 mol/L.

In one embodiment, the column is ZORBZX Eclipse XDB-C18analytical 4.6X 150mm 5-Micron or Kromasil 100-5-C184.6X 150 mm.

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

In one embodiment, the chlorhexidine gluconate content in the test solution is 0.05 mg/mL-0.15 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:

and carrying out HPLC detection by using a chlorhexidine gluconate reference substance according to an external standard method to obtain the relation between the content of the chlorhexidine gluconate and the peak area, and calculating 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.

In one embodiment, a standard curve is drawn by using a chlorhexidine gluconate reference substance according to an external standard method, and the content of the chlorhexidine gluconate in the sample to be detected is calculated according to the standard curve.

Specifically, a proper amount of chlorhexidine gluconate reference substance is taken, a chlorhexidine gluconate reference sample containing 0.02 mg/mL-0.2 mg/mL is prepared in a solvent with the same components as the mobile phase, sample treatment and detection are carried out in parallel with the test sample solution, the concentration of the chlorhexidine gluconate corresponding to each peak area is measured, and a standard curve is drawn;

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

According to the detection method, the appropriate mobile phase component is selected, the ratio composition of the mobile phase is adjusted, and other parameter conditions are combined, so that the response value of the detection condition is high, the chlorhexidine gluconate has a fast peak under the condition that the separation degree of the components is ensured, the retention time of the chlorhexidine gluconate can be effectively shortened, the content of the chlorhexidine gluconate in a shampoo preparation containing the chlorhexidine gluconate can be quickly and accurately determined, the problem of determining the content of the chlorhexidine gluconate in the compound miconazole nitrate shampoo is solved, the detection working efficiency is greatly improved, and the quality of a product can be favorably controlled. In addition, the detection method can be applied to shampoo preparations containing chlorhexidine gluconate and can also be applied to content detection of the chlorhexidine gluconate in other products or preparations.

Drawings

FIG. 1 is an HPLC detection spectrum of chlorhexidine gluconate in compound miconazole nitrate shampoo of example 1;

FIG. 2 is the HPLC detection profile 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, various additives and the like; chlorhexidine gluconate control was purchased from Sigma; acetonitrile and triethylamine were chromatographically pure, and disodium hydrogen phosphate and phosphoric acid were analytically pure.