CN117890506A - Method for detecting chlorpheniramine maleate content in cosmetics - Google Patents

Method for detecting chlorpheniramine maleate content in cosmetics Download PDF

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Publication number
CN117890506A
CN117890506A CN202311853019.6A CN202311853019A CN117890506A CN 117890506 A CN117890506 A CN 117890506A CN 202311853019 A CN202311853019 A CN 202311853019A CN 117890506 A CN117890506 A CN 117890506A
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chlorpheniramine maleate
cosmetics
content
solution
performance liquid
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马驰远
段吴燕
马欣欣
占春瑞
万建春
李仕祥
王佳
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Nanchang Customs Technical Center
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/04Preparation or injection of sample to be analysed
    • G01N30/06Preparation
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/62Detectors specially adapted therefor
    • G01N30/74Optical detectors
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/86Signal analysis
    • G01N30/8675Evaluation, i.e. decoding of the signal into analytical information
    • G01N30/8679Target compound analysis, i.e. whereby a limited number of peaks is analysed
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/04Preparation or injection of sample to be analysed
    • G01N30/06Preparation
    • G01N2030/065Preparation using different phases to separate parts of sample

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  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
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  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
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  • Engineering & Computer Science (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Cosmetics (AREA)

Abstract

The invention relates to a method for detecting chlorpheniramine maleate content in cosmetics, which specifically selects different extraction solvents for pretreatment aiming at different dosage forms of cosmetics, and the obtained test solution is used for measuring the chlorpheniramine maleate content by a high performance liquid chromatograph. Wherein, the cosmetics in the dosage forms of water agent, gel, cream, emulsion, film and powder adopt methanol as extraction solvent; the method can effectively solve the problem that hydrophilic target substances to be detected in cosmetics are difficult to effectively extract from a hydrophobic matrix. The method is quick, simple and convenient, has high sensitivity, the average recovery rate of chlorpheniramine maleate in actual cosmetic detection is 85.3-104.9%, the relative standard deviation is 1.54-9.42%, and the accuracy and the repeatability are good, so that technical support can be provided for cosmetic supervision.

Description

Method for detecting chlorpheniramine maleate content in cosmetics
Technical Field
The invention relates to a method for detecting chlorpheniramine maleate content in cosmetics, and particularly belongs to the technical field of liquid chromatography analysis.
Background
Cosmetics have been incorporated into people's daily lives, and skin diseases caused by adverse reactions of skin and its appendages have been greatly increased with widespread use, skin allergy being one of the most common. At present, the addition of anti-sensitizers to cosmetics is not secret. Chlorpheniramine maleate is a representative antiallergic drug, and has a certain inhibition effect on the central nervous system, so that the chlorpheniramine maleate is clinically mainly used for treating skin allergic urticaria and drug eruptions caused by drug allergy, but the use of the drug can cause side effects such as physical strength decline, hypodynamia, somnolence and the like. The national food and drug administration issues "technical Specification for safety of cosmetics" (2015 edition) to prescribe high performance liquid chromatography-tandem mass spectrometry to measure the content of chlorpheniramine in cosmetics, but the method cannot effectively extract components to be measured in film-sticking, powder, oil and wax-based cosmetic samples, and strong solvents are selected as extraction solvents to easily erode instrument sealing elements, so that instrument maintenance cost is increased, and market popularization difficulty is high. At present, research on the use limit of chlorpheniramine maleate in cosmetics at home and abroad has not been decided, and related reports on high performance liquid chromatography detection of chlorpheniramine maleate in medicines are available. In the drug detection, chlorpheniramine maleate has better hydrophilicity, so the sample pretreatment extraction solvent is hydrophilic organic solution, but the cosmetic matrix dosage form mainly comprises 8 types of aqueous agents, gels, creams, emulsions, films, powders, oil agents and wax bases, and the existing detection pretreatment method of the cosmetics is difficult to effectively extract hydrophilic target objects to be detected from a hydrophobic matrix, so the detection method for effectively extracting chlorpheniramine maleate from various cosmetics is lacked. Therefore, it is necessary to research and establish a method for detecting chlorpheniramine maleate content of 8 dosage forms of cosmetics such as water agent, gel, cream, emulsion, film, powder, oil agent and wax base, which has simple sample pretreatment, low cost and high instrument popularization rate.
At present, high performance liquid chromatography detection and analysis methods for measuring the chlorpheniramine maleate content in large-scale cosmetics such as water agents, gels, creams, emulsions, films, powders, oil agents and wax-based 8 are not available at home and abroad. Therefore, according to the physicochemical properties of chlorpheniramine maleate, the invention adopts different extraction solvents for different formulations of cosmetics, and the content of chlorpheniramine maleate is measured accurately and reliably by optimizing high performance liquid chromatography conditions, thereby providing technical support for further strengthening the supervision of cosmetics.
Disclosure of Invention
The invention provides a method for detecting chlorpheniramine maleate content in cosmetics, aiming at the defects of the prior art.
According to the method for detecting chlorpheniramine maleate content in cosmetics, different extraction solvents are selected for pretreatment aiming at cosmetics of different dosage forms, and the obtained test solution is used for measuring the chlorpheniramine maleate content by using a high performance liquid chromatograph;
the method comprises the following specific steps:
step 1: preparing test solution
The cosmetics in the dosage forms of water aqua, gel, cream, emulsion, film and powder adopt methanol as extraction solvent, and the content of chlorpheniramine maleate in the extracted test solution is measured by a high performance liquid chromatograph;
The oil agent and wax-based cosmetics sequentially adopt acetonitrile saturated normal hexane solution and normal hexane saturated acetonitrile solution as extraction solvents, and the extracted test solution is used for measuring the content of chlorpheniramine maleate by a high performance liquid chromatograph;
step 2: preparing standard working solution
Methanol is used as a solvent, and chlorpheniramine maleate standard substances are prepared into standard working solution with the concentration of 1.0-50.0 mug/mL;
step 3: high performance liquid chromatograph for measuring content of chlorpheniramine maleate
Separating the sample solution by a reversed-phase Agilent Zorbox SB-C 18 chromatographic column after filtering, taking phosphate buffer solution and acetonitrile as mobile phases, and adopting a high-performance liquid chromatograph to quantitatively analyze and detect the content of chlorpheniramine maleate by an external standard method, wherein the flow rate is 1-1.5 mL/min, the column temperature is 30-35 ℃, the detection wavelength is 262-nm and the sample injection amount is 5-20 mu L.
The phosphate buffer solution is prepared by dissolving 11.5g of monoammonium phosphate with pure water, adding 1mL of phosphoric acid to adjust the pH value to 2.5, and fixing the volume to 1L with the pure water.
The volume ratio of the phosphate buffer solution to the acetonitrile in the mobile phase is 80:20.
The invention has the beneficial effects that: 1. the invention establishes a detection method suitable for measuring the chlorpheniramine maleate content in various dosage forms of cosmetics, has the advantages of simple pretreatment method of cosmetic samples, low toxicity of extraction solvents, accurate quantitative qualitative, quick and efficient detection, low detection cost, strong market popularization of the adopted high performance liquid chromatograph, suitability for detecting the chlorpheniramine maleate content in various dosage forms of cosmetics, and provides technical support for further strengthening the supervision of cosmetics.
2. The invention solves the problem that hydrophilic target substances to be detected in cosmetics are difficult to effectively extract from a hydrophobic matrix, and aims at oil and wax-based formulation cosmetic samples, the oil and wax-based formulation cosmetic samples are effectively prevented from being dissolved out and have a purification effect by adopting acetonitrile saturated n-hexane solution for water bath ultrasonic dissolution, then adding n-hexane saturated acetonitrile solution for rapid discrete liquid-liquid extraction, effectively separating the target substances to be detected into lower-layer n-hexane saturated acetonitrile solution, and carrying out solvent replacement.
3. According to the invention, through full-wavelength scanning of chlorpheniramine maleate, the maximum absorption wavelength of chlorpheniramine maleate is determined, interference of an extraction solvent and impurities in a sample is avoided, a pretreatment method of the sample is improved, a fluidity system is inspected, a detection method for measuring the content of chlorpheniramine maleate in various dosage forms of cosmetics is established, the accuracy and reproducibility are good, the average recovery rate of chlorpheniramine maleate in actual sample detection is 85.3% -104.9%, and the relative standard deviation is 1.54% -9.42%, so that the invention can provide reference for effectively extracting other hydrophilic targets to be detected in cosmetics.
Drawings
Fig. 1A: the chlorpheniramine maleate full-wavelength scanning 3D image is provided;
fig. 1B: the chlorpheniramine maleate and other absorption patterns of the invention;
Fig. 2: according to the invention, agilent Zorbox SB-C 18 chromatographic columns are selected for separating chlorpheniramine maleate standard substance chromatograms;
fig. 3: chromatographic separation and response profiles of flow versus chlorpheniramine maleate;
fig. 4: linear range, regression equation and linear relation coefficient of chlorpheniramine maleate;
In the figure: chlorpheniramine maleate has a linear range of 1.0-50.0 mug/mL, a regression equation y= 4.49510x-4.00589, a linear correlation coefficient of 0.99921, a retention time of 11.968min and a residual standard error of 3.49870.
Detailed Description
The invention is further described below with reference to examples.
Example 1
Pretreatment of aqueous, gel, cream, emulsion, film and powder cosmetic samples containing chlorpheniramine maleate:
Weighing 1.00g of the cosmetic sample, placing the cosmetic sample in a 100mL colorimetric tube, adding 20mL of methanol, mixing by vortex, carrying out ultrasonic extraction for 10min in a water bath at 50 ℃, cooling to room temperature, fixing the volume to 50mL by using the methanol, mixing uniformly, transferring 10mL of sample liquid into a 50mL centrifuge tube, centrifuging for 5min at 4500 r/min, taking supernatant, and filtering with a 0.45 mu m filter membrane, wherein the obtained test liquid is used for measuring the content of chlorpheniramine maleate by a liquid chromatograph.
Example 2
Pretreatment of oil and wax-based cosmetic samples containing chlorpheniramine maleate:
1.00g of the cosmetic sample is weighed and placed in a 100mL colorimetric tube, 15mL of acetonitrile saturated normal hexane solution is added, vortex is carried out for 1min, ultrasonic extraction is carried out for 20min in a water bath at 60 ℃, vortex is carried out for 1min, cooling is carried out to room temperature, then 50mL of normal hexane saturated acetonitrile solution is added, vortex mixing is carried out for 3min, the upper solution is removed, 5mL of lower solution is removed, the lower solution is placed in a 15mL centrifuge tube, nitrogen is blown to dryness, 5mL of methanol is added for redissolution, 4500 r/min is carried out for 5min, supernatant is taken and filtered through a 0.45 mu m filter membrane, and the obtained test solution is used for measuring the content of chlorpheniramine maleate by a liquid chromatograph.
Example 3
The method for detecting and analyzing the chlorpheniramine maleate content in the cosmetic sample by using a high performance liquid chromatograph.
The implementation steps are as follows:
step 1: detection of the optimal wavelength of chlorpheniramine maleate
Chlorpheniramine maleate was scanned across the 190-450nm full band by a Diode Array Detector (DAD) with the best response at 262 nm wavelength as shown in fig. 1A and 1B for the maximum absorption wavelength results. Thus, this example determines that the detection optimum wavelength for chlorpheniramine maleate is set to 262 nm.
Step 2: c 18 chromatographic column for detecting chlorpheniramine maleate
The chlorpheniramine maleate is separated by selecting Agilent Zorbox SB-C 18 chromatographic column (5 μm, 4.6X1250 mm), the chromatogram is shown in figure 2, and when Agilent Zorbox SB-C 18 (5 μm, 4.6X1250 mm) chromatographic column is selected as C 18 chromatographic column for detecting chlorpheniramine maleate content, the chromatographic peak separation effect is good. Thus, this example identifies the C 18 column for detecting chlorpheniramine maleate content as Agilent Zorbox SB-C 18 (5 μm, 4.6X1250 mm) column.
Step 3: detection of the Mobile phase of chlorpheniramine maleate
The method takes ammonium dihydrogen phosphate buffer salt solution as a mobile phase, researches the chromatographic separation and response effects of the flow relative to chlorpheniramine maleate by two basic parameters of peak area, peak height, neutralization tower plate number and symmetry factor, and the result is shown in figure 3, and the ionization form can be well controlled by taking ammonium dihydrogen phosphate buffer solution as the mobile phase for detecting chlorpheniramine maleate, so that the chromatographic peak of chlorpheniramine maleate is ensured to be free from tailing, branching and deformation.
Example 4
The method for detecting the chlorpheniramine maleate content in various cosmetics is verified in methodology.
1. Main reagent and instrument
Chlorpheniramine maleate (CAS: 113-92-8), standard, dr. Ehrenstorfer, germany; methanol, acetonitrile, n-hexane and chloroform, chromatographically pure, merck, inc; phosphoric acid, analytically pure, chemical company, inc; triethylamine, analytically pure, adult western asia chemical company, inc; monoammonium phosphate, analytically pure, national pharmaceutical group chemical reagent company, inc.
Agilent 1260 high performance liquid chromatograph, diode Array Detector (DAD), ultraviolet detector (VWD), agilent technologies, inc; neofuge1600R centrifuge, shanghai Lishen scientific instruments Co., ltd; VORTE 3A vortex oscillator, germany Ai Ka instrument, inc.; SB25-12DT ultrasonic cleaner, ningbo Xinzhi biotechnology Co., ltd.
2. Preparation of standard working solution
Accurately weighing 10 mg (accurate to 0.1 mg) chlorpheniramine maleate standard substance, dissolving in 10 mL volumetric flask, and fixing volume to scale with methanol to obtain 1.0 mg/mL standard stock solution, and storing at 4 deg.C in dark place. Accurately sucking the standard stock solution, preparing 100 mug/mL standard intermediate solution by using methanol, and respectively sucking the standard intermediate solution to prepare standard working solution with the concentration of 1.0-50.0 mug/mL.
3. Preparation of extraction solvent and buffer solution
Respectively taking 200mL of normal hexane and acetonitrile solution, and carrying out ultrasonic mixing in a 1L reagent bottle to obtain an extraction solvent consisting of an upper acetonitrile saturated normal hexane solution and a lower normal hexane saturated acetonitrile solution.
11.5G of monoammonium phosphate is weighed and dissolved by pure water, 1mL of phosphoric acid is added to adjust the pH value to 2.5, and the volume is fixed to 1L by pure water, so as to obtain the buffer salt solution of monoammonium phosphate.
4. Pretreatment of cosmetic samples:
Pretreatment was performed in the manner of example 1 and example 2.
5. Determining chromatographic conditions:
The chromatographic column is Agilent Zorbox SB-C 18 chromatographic column (5 μm, 4.6X1250 mm); the mobile phase is phosphate buffer solution-acetonitrile, (80:20, V/V); the flow rate is 1.2 mL/min; column temperature is 30 ℃; wavelength 262 nm; the sample injection amount was 10. Mu.L.
6. Linear relation of method, detection limit and quantitative limit:
And drawing a standard curve by taking the concentration of the standard working solution as an abscissa and the peak area of the chromatographic peak of the target object to be detected as an ordinate, so as to obtain the standard curve and the correlation coefficient. As shown in FIG. 4, when chlorpheniramine maleate is in the range of 1.0-50.0 mug/mL, the linearity is good, and the correlation coefficient is 0.99921. The detection limit and the quantitative limit of the embodiment are determined by adopting 20 groups of blank repeated measurement, the detection limit is calculated by 3 times of signal to noise ratio, the quantitative limit is calculated by 10 times of signal to noise ratio, and the detection limit of chlorpheniramine maleate is determined to be 15 mug/g and the quantitative limit is determined to be 50 mug/g.
7. And (3) adding standard recovery rate and precision:
In this example, samples of common 8 types of cosmetic formulations are used as a matrix, and 3-level 6-parallel labeling recovery experiments are performed on the samples by using a standard adding method, wherein the adding concentrations are respectively 1-fold, 2-fold and 10-fold quantitative limits, and the results are shown in table 1.
As can be seen from Table 1, the recovery rate of chlorpheniramine maleate with 1 time, 2 times and 10 times of quantitative limit is 85.3% -104.9% and the relative standard deviation is 1.54% -9.42% (n=6) by taking the skin cream as a cosmetic sample, which shows that the accuracy and repeatability are good and the related detection requirements can be met.
Example 5
This example tested 33 cosmetic actual samples of different brands for class 8 common cosmetic formulations.
Wherein, 1.00g of the cosmetic samples of water agent, gel, cream, emulsion, film and powder are weighed and placed in a 100mL colorimetric tube, 20mL of methanol is added, the mixture is uniformly mixed by vortex, the mixture is subjected to ultrasonic extraction in water bath at 50 ℃ for 10min, cooled to room temperature, the volume is fixed to 50mL by the methanol, the mixture is uniformly mixed, 10mL of sample liquid is removed in a 50mL centrifuge tube, the mixture is centrifuged for 5min at 4500 r/min, and the supernatant is filtered by a 0.45 mu m filter membrane and detected by a high performance liquid chromatograph.
Wherein, 1.00g of oil and wax-based cosmetic sample is weighed and placed in a 100mL colorimetric tube, 15mL of acetonitrile saturated normal hexane solution is added, 1min is vortexed, the ultrasonic extraction is carried out in a 60 ℃ water bath for 20min, 1min is vortexed, the cooling is carried out to room temperature, 50mL of normal hexane saturated acetonitrile solution is added, the vortexed mixing is carried out for 3min, the upper solution is discarded, 5mL of lower solution is removed in a15 mL centrifuge tube, nitrogen is blown to dryness, 5mL of methanol is added for redissolution, 4500 r/min is carried out for 5min, the supernatant is taken to pass through a 0.45 mu m filter membrane, and the detection is carried out through a high performance liquid chromatograph.
The high performance liquid chromatography instrument method adopts Agilent Zorbox SB-C 18 chromatographic column, the mobile phase is isocratic monoammonium phosphate buffer salt-acetonitrile solution (80:20, V/V), the detection wavelength is 262 nm, the flow rate is 1.2 mL/min, the column temperature is 30 ℃, and the sample injection amount is 10 mu L.
The test results are shown in Table 2.
As can be seen from Table 2, chlorpheniramine maleate is detected in 4 kinds of sun-proof cosmetics, and one of the domestic sun-proof cream chlorpheniramine maleate is detected as undetected, the chlorpheniramine maleate imported from the same brand and same type of product is 3.1mg/g, and the invention establishes a detection method for measuring the content of chlorpheniramine maleate in various types of cosmetics, which can provide technical support for detecting import and export cosmetics.

Claims (3)

1. A method for detecting chlorpheniramine maleate content in cosmetics is characterized in that: according to the method, different extraction solvents are selected for pretreatment aiming at cosmetics with different dosage forms, and the obtained test solution is used for measuring the content of chlorpheniramine maleate by a high performance liquid chromatograph;
the method comprises the following specific steps:
step 1: preparing test solution
The cosmetics in the dosage forms of water aqua, gel, cream, emulsion, film and powder adopt methanol as extraction solvent, and the extracted test solution is used in high performance liquid chromatograph to determine chlorpheniramine maleate content;
The oil agent and wax-based cosmetics sequentially adopt acetonitrile saturated normal hexane solution and normal hexane saturated acetonitrile solution as extraction solvents, and the extracted test solution is used for measuring the content of chlorpheniramine maleate by a high performance liquid chromatograph;
step 2: preparing standard working solution
Methanol is used as a solvent, and chlorpheniramine maleate standard substances are prepared into standard working solution with the concentration of 1.0-50.0 mug/mL;
step 3: high performance liquid chromatograph for measuring content of chlorpheniramine maleate
Separating the sample solution by a reversed-phase Agilent Zorbox SB-C 18 chromatographic column after filtering, taking phosphate buffer solution and acetonitrile as mobile phases, and adopting a high-performance liquid chromatograph to quantitatively analyze and detect the content of chlorpheniramine maleate by an external standard method, wherein the flow rate is 1-1.5 mL/min, the column temperature is 30-35 ℃, the detection wavelength is 262-nm and the sample injection amount is 5-20 mu L.
2. The method for detecting chlorpheniramine maleate content in cosmetics according to claim 1, wherein the method comprises the following steps: the phosphate buffer solution is prepared by dissolving 11.5g of monoammonium phosphate with pure water, adding 1mL of phosphoric acid to adjust the pH value to 2.5, and fixing the volume to 1L with the pure water.
3. The method for detecting chlorpheniramine maleate content in cosmetics according to claim 1, wherein the method comprises the following steps: the volume ratio of the phosphate buffer solution to the acetonitrile in the mobile phase is 80:20.
CN202311853019.6A 2023-12-29 2023-12-29 Method for detecting chlorpheniramine maleate content in cosmetics Pending CN117890506A (en)

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