CN102735791A - Quality detection method for antifungal cream - Google Patents

Abstract

A quality detection method for Funing cream, comprising the identification of the antifungal active ingredient econazole nitrate and the determination of its content. The determination of the content of econazole nitrate is to use a third-order reciprocal spectroscopic method. First, separate and extract the peptide For econazole nitrate in Ning cream, accurately weigh about 1g of Funing cream to be tested, add 15ml of solvent, the solvent is made by mixing 0.1mol/L HCL and methanol at a volume ratio of 1:9, in a water bath of 80 Heat and stir at ℃ until clarified, put in the refrigerator to cool to below room temperature, filter and wash three times, use 10ml of the solvent each time, combine the filtrate into a 50ml volumetric flask, add the solvent to dilute to the mark, shake well, and measure accurately Put 2.5ml of the diluent in a 25ml volumetric flask, add solvent to the mark, and shake up; secondly, use a UV-visible spectrophotometer to measure the econazole nitrate content at 231nm with the third-order derivative method. The detection method of the invention is accurate, has good specificity and repeatability, is convenient to operate, and has low cost.

Description

A kind of quality detection method of antifungal cream

technical field

The invention relates to a quality detection method of cream preparation, in particular to a quality detection method of antifungal cream.

Background technique

Because skin infections are mostly fungal infections, often accompanied by bacterial infections ^[1] , it is difficult to achieve therapeutic effect with a single antifungal drug. According to this feature, the antifungal drug econazole nitrate, aminoglycoside antibiotics, gentamicin sulfate, and Funing cream compatible with hormone dexamethasone sodium phosphate are often used for skin infections. Skin itching embolism. The prescription of Funing cream is: econazole nitrate 10g, gentamicin sulfate 10g, dexamethasone sodium phosphate 0.6g, solution A (stearic acid 60g, Span 6016g, stearyl alcohol 60g, liquid paraffin 90g, white Petroleum jelly 60g), B solution (polysorbate 8044g, glycerin 100g, sorbic acid 2g, add distilled water to 1000g.), heat A and B in a water bath to about 80°C, slowly add B solution to A under stirring Stir while adding, take out after mixing, continue to stir and cool to about 50°C, add econazole nitrate, gentamicin sulfate, dexamethasone sodium phosphate, etc. in proportion, continue to stir until condensation, that is have to. There are many manufacturers producing Funing Cream, but Funing Cream is an antifungal drug used in clinical practice, and it is necessary to formulate feasible quality control methods to ensure product quality and clinical efficacy.

The analytical method that uses the principles and experimental methods of spectroscopy to determine the structure and chemical composition of substances is called spectroscopic analysis. Substances of various structures have their own characteristic spectra. Spectral analysis is a method of using characteristic spectra to study the structure of substances or determine the chemical composition. UV-Vis absorption spectroscopy is to study the absorption spectrum of molecules absorbing in the wavelength range of 190-750nm. The ultraviolet-visible absorption spectrum is mainly produced by the transition of valence electrons in molecules between electronic energy levels. It is an analytical method for studying the electronic spectrum of substances. By measuring the absorption of ultraviolet-visible light by molecules, a large number of inorganic compounds and organic compounds can be identified and measured. The molecular absorption spectrum produced by a substance absorbing electromagnetic radiation energy in the wavelength range of 200-760nm is called the ultraviolet-visible absorption spectrum of the substance, and the qualitative and quantitative analysis method of the substance using the ultraviolet-visible absorption spectrum is called ultraviolet - Visible spectrophotometry. Derivative spectrophotometry is to perform differential processing on the usual absorption spectrum, calculate the value of △A/△λ point by point every other small wavelength interval △λ (generally 1-2nm), and draw the △A/△λ～λ curve , the resulting graph is the first derivative spectrum. In turn, the higher-order derivative spectrum can be obtained. With the increase of the derivative order, the number of peaks increases, the peak width narrows, and the resolving power improves, which is beneficial to the separation and identification of overlapping bands and shoulder peaks. The use of derivative spectrophotometry can eliminate the interference absorption of coexisting components, and can also improve the sensitivity of the determination. At present, the use of a spectrophotometer with a microprocessor can automatically calculate and directly describe the first-order, second-order, third-order, fourth-order and other derivative spectra.

At present, high-performance liquid phase HPLC is mostly used for drug quality control, but HPLC is expensive and difficult to maintain. Compared with HPLC, ultraviolet spectrometers are cheap and easy to operate. Currently, there is no non-HPLC method for the detection of complex pharmaceutical ingredients such as econazole nitrate in creams.

Contents of the invention

The object of the present invention is to provide a quality detection method of the antifungal drug Funing Cream, which is simple to operate and low in cost, and can effectively guarantee the quality of the Funing Cream and its antifungal clinical curative effect.

The object of the invention is achieved through the following technical measures:

A quality detection method for Funing cream, comprising the identification of the antifungal active ingredient econazole nitrate and the determination of its content, characterized in that: the identification of econazole nitrate adopts a thin-layer chromatography identification method, specifically Take 2g of Funing cream to be tested, 20ml of methanol, heat and stir in a water bath at 80°C, cool in an ice bath, filter, evaporate the filtrate to dryness, add 2ml of methanol to dissolve it as the test solution, accurately weigh 20mg of econazole nitrate sample, dissolve it in 2ml of methanol As a control solution, take 5ul points on a silica gel plate, develop with ether, and develop color with iodine vapor for comparison;

The determination of the econazole nitrate content is to adopt the third-order reciprocal spectroscopic method, first separate and extract the econazole nitrate in the Funing cream, accurately weigh about 1g of the Funing cream to be tested, add 15ml of solvent, and The solvent is mixed with 0.1mol/L HCL and methanol at a volume ratio of 1:9, heated and stirred in a water bath at 80°C until clarified, cooled in a refrigerator to below room temperature, filtered, washed three times, each time with 10ml of the solvent, Merge the filtrate into a 50ml volumetric flask, add the solvent to dilute to the mark, shake up, accurately measure 2.5ml of the diluent in a 25ml volumetric flask, add the solvent to the mark, and shake up; secondly, use a UV-visible spectrophotometer , Determination of econazole nitrate content at 231nm by third order derivative method.

The quality detection method of the above-mentioned Funing cream also includes identifying the antibacterial active ingredient gentamicin sulfate, specifically taking 0.5g of Funing cream to be tested (containing 5 mg of gentamicin sulfate) and dissolving it in 10ml of water, and boiling water bath to dissolve milk, add 0.1wt% (mass percent concentration) ninhydrin water-saturated n-butanol solution 1ml and pyridine 0.5ml while hot, and heat in a water bath at 100°C for 5min to develop color.

The quality detection method of the above-mentioned Funing cream also includes identifying dexamethasone sodium phosphate, specifically take 10 g of Funing cream to be tested (containing 6 mg of dexamethasone sodium phosphate), add 10 ml of methanol, and dissolve it in a water bath at 80 ° C. After hot filtration, the filtrate was cooled and evaporated to dryness with a rotary evaporator, and the residue was added to 1ml sulfuric acid for color development.

Specifically, a quality detection method for Funing cream is the identification of econazole nitrate and the determination of its content, the identification of gentamicin sulfate as an active ingredient against bacteria, and the identification of dexamethasone phosphate The sodium component is identified; it is characterized in that: the identification of econazole nitrate adopts thin-layer chromatography identification method, specifically taking 2 g of Funing cream to be tested, 20 ml of methanol, heating and stirring in a water bath at 80 ° C, cooling in an ice bath, and filtering , the filtrate was evaporated to dryness, added 2ml of methanol to dissolve as the test solution, accurately weighed 20mg of econazole nitrate sample, 2ml of methanol was dissolved as the reference solution, divided 5ul points on the silica gel plate, developed with ether, and developed color with iodine vapor for comparison; The determination of the econazole nitrate content is to adopt the third-order reciprocal spectroscopic method, first separate and extract the econazole nitrate in the Funing cream, accurately weigh about 1g of the Funing cream to be tested, add 15ml of solvent, and The solvent is mixed with 0.1mol/L HCL and methanol at a volume ratio of 1:9, heated and stirred in a water bath at 80°C until clarified, cooled in a refrigerator to below room temperature, filtered, washed three times, each time with 10ml of the solvent, Merge the filtrate into a 50ml volumetric flask, add the solvent to dilute to the mark, shake up, accurately measure 2.5ml of the diluent in a 25ml volumetric flask, add the solvent to the mark, and shake up; secondly, use a UV-visible spectrophotometer , Determination of econazole nitrate content with the third order derivative method at 231nm;

To identify gentamicin sulfate, specifically, take 0.5g of Funing cream to be tested (containing 5mg of gentamicin sulfate) and dissolve it in 10ml of water, break the emulsion in a boiling water bath, and add 0.1wt% (mass percentage concentration) indene while it is hot 1ml of water-saturated n-butanol solution of triketone and 0.5ml of pyridine were heated in a water bath at 100°C for 5min to develop color;

Dexamethasone sodium phosphate was identified by taking 10g of Funing cream to be tested (containing 6mg of dexamethasone sodium phosphate), adding 10ml of methanol, dissolving in 80°C water bath, filtering while hot, and evaporating to dryness with a rotary evaporator after the filtrate was cooled. , the residue was added to 1ml sulfuric acid for color development.

The present invention has following beneficial effect:

The quality detection method of the antifungal Funing cream preparation of the present invention is easy to operate and low in cost, and it can easily and quickly identify the antifungal active ingredient econazole nitrate and its content, especially in the determination of econazole nitrate Methanol is used as the extraction solvent, which fully reduces the interference of impurities in the liquid to be tested. It is developed in ether, and the effect is very good, and no impurity spots appear. In the determination of econazole nitrate content, a specific solvent can be used to break the emulsion and extract econazole nitrate. The detection method of the present invention has a high recovery rate, which is more than 99% each time, the average recovery rate is 100.3%, and the reproducibility is good; at the same time, the accuracy is high, and the inventive method r=0.9999 is approximately 1, which is comparable to the HPLC method. Equivalent to accuracy. In a word, the detection method of the present invention is accurate, has good specificity and repeatability, is convenient to operate, and has low cost. The present invention provides a simple alternative method for many schools, hospitals and small laboratories that do not have the conditions.

Description of drawings

Figure 1 is the ultraviolet zero-order spectrum, a econazole nitrate b Funing cream c blank matrix; the unit wave number (nm) of the abscissa is 200.0, 220.0, 240.0, 260.0, 280.0, 300.0, and the ordinate is the amplitude, They are -0.025, 1.000, 2.000, 2.500 respectively.

Figure 2 is the third derivative spectrum of econazole nitrate, a econazole nitrate b Funing cream c blank matrix; abscissa unit wave number (nm), respectively 200.0, 220.0, 240.0, 260.0, 280.0, 300.0, vertical The coordinates are the derivatives of the amplitude, which are -0.025, -0.020, -0.010, 0.000, 0.010, 0.020, 0.025, respectively.

Detailed ways

The present invention is specifically described below through the examples, it is necessary to point out that the following examples are only used to further illustrate the present invention, and can not be interpreted as limiting the protection scope of the present invention, those skilled in the art can according to the above-mentioned present invention Contents Some non-essential improvements and adjustments are made to the present invention.

Example 1

A kind of quality detection method of Funing emulsifiable paste, carry out according to the following steps:

(1) Identification of econazole nitrate by thin-layer chromatography: take 2g of Funing cream sample, 20ml of methanol, heat and stir in a water bath at 80°C, cool in an ice bath, filter, evaporate the filtrate to dryness, add 2ml of methanol to dissolve it as a test, and weigh it accurately Dissolve 20mg of econazole nitrate sample in 2ml of methanol as a control solution, take 5ul and spot it on a silica gel plate, develop with ether, and develop color with iodine vapor. The main spots of the control solution and the test solution are at the same position;

(2) Differentiate gentamicin sulfate: get Funing emulsifiable paste sample 0.5g (equivalent to gentamicin sulfate 5mg) and add water 10ml to dissolve, and the boiling water bath breaks the emulsification, adds the water-saturated normal of 0.1wt% ninhydrin while hot 1ml of butanol solution and 0.5ml of pyridine, heated in a water bath at 100°C for 5min, the solution will appear blue-purple;

(3) Identification of dexamethasone sodium phosphate: Take 10g of sample (equivalent to 6mg of dexamethasone sodium phosphate), add 10ml of methanol, dissolve in 80°C water bath, filter while hot, evaporate the filtrate to dryness with a rotary evaporator after cooling, add 1ml of residue Sulfuric acid was brownish red.

(4) Three-order reciprocal spectrometry measures econazole nitrate content:

1. Solution Preparation

Preparation of the test solution: Accurately weigh about 1g of the sample, add 15ml of solvent (0.1mol/L HCL-methanol 1:9, in volume ratio), heat and stir in a water bath at 80°C until clear, and cool in a refrigerator to below Filter and wash at room temperature three times, each time with 10ml of solvent, combine the filtrate into a 50ml volumetric flask, add solvent to the mark, shake well, accurately measure 2.5ml of the diluent into a 25ml volumetric flask, add solvent to the mark, and shake well.

Preparation of blank control solution: prepare a blank sample solution not containing econazole nitrate according to the preparation method of the above-mentioned need testing solution.

Preparation of the reference solution: Accurately weigh 10.2mg of econazole nitrate, dissolve and dilute it to a 50ml volumetric flask with a solution of 0.1mol/L HCL-methanol 1:9 (volume ratio), and accurately measure 2.5ml of the diluted solution to 25ml volumetric flask.

2. Spectral conditions and system suitability experiments

Instruments and reagents: UV-2501PC (Shimadzu, Japan), analytical pure methanol, n-butanol, hydrochloric acid. Fu Ning Cream (produced by the Pharmacy Department of Xinqiao Hospital). Econazole nitrate (batch number 030208, Xuzhou Nhua Group).

3. Assay

3.1 Selection of experimental conditions

Preparation of the test solution: Accurately weigh about 1g of the sample, add 15ml of solvent (0.1mol/L HCL-methanol 1:9), heat and stir in a water bath at 80°C until clear, cool in the refrigerator to below room temperature, filter and wash three times , 10ml of solvent each time, combine the filtrate into a 50ml volumetric flask, add solvent to the mark, shake well, accurately measure 2.5ml of diluent in a 25ml volumetric flask, add solvent to the mark, and shake well.

Preparation of control solution: Accurately weigh 10.2mg of econazole nitrate, dissolve it with 0.1mol/L HCL-methanol 1:9 solution according to law and dilute it into a 50ml volumetric flask, and accurately measure 2.5ml of the diluent into a 25ml volumetric flask.

Preparation of blank solution: Accurately weigh about 1g of cream without econazole nitrate, add 15ml of solvent (0.1mol/L HCL-methanol 1:9), heat and stir in a water bath at 80°C and cool in a refrigerator to below room temperature. Filter and wash three times, 10ml of solvent each time, set the volume in a 50ml volumetric flask at room temperature, accurately measure 2.5ml of the diluent in a 25ml volumetric flask, and shake well.

3.2 Selection of measurement wavelength The test solution, control solution and blank solution respectively use 0.1mol/L hydrochloric acid-methanol as a blank, and scan in the wavelength range of 300~200nm to obtain zero-order and third-order spectra, see Figure 1 and Figure 2 . As can be seen from Figure 1, in the zero-order ultraviolet spectrum, the blank matrix interferes greatly with the absorption of econazole nitrate. The third-order derivative spectrogram has the maximum amplitude at 231nm of econazole nitrate, and the blank matrix has almost no amplitude value, which can exclude the influence of the blank matrix. Therefore, the content of econazole nitrate is determined by the third-order derivative method at 231nm.

3.3 Drawing of standard curve

Accurately weigh 10.2mg of econazole nitrate dried to constant weight, dissolve in 0.1mol/L hydrochloric acid-methanol (1:9) solution, set the volume in a 50ml volumetric flask, and accurately measure diluents 1.5ml, 2ml, 2.5ml respectively ml, 3ml, 3.5ml, set the volume to 25ml volumetric flask, and prepare solutions with concentrations of 12.24ug/ml, 16.32ug/ml, 20.4ug/m2, 24.48ug/ml, 28.56ug/ml, and measure three at 231nm The spectral amplitude value of the first derivative, the concentration has a very good linear relationship with the third derivative value C=(D-0.00010)/0.000221 r=0.9999 is approximately 1.

3.4 Determination of recovery rate

Accurately weigh several portions of econazole nitrate with different concentrations, add it into 1g of blank cream that does not contain econazole nitrate, heat, mix and cool, cool until the cream is formed, add 15ml of solvent (0.1mol/L HCL-methanol 1:9), heat and stir in a water bath at 80°C until clear, take it out, put it in the refrigerator, cool it to room temperature, filter, wash three times (pick out the filtered matrix and re-emulsify with solvent for each time), 10ml of solvent each time, Combine the filtrates, set the volume at room temperature in a 50ml volumetric flask, accurately measure 2.5ml of diluent, add solvent to the 25ml volumetric flask to the mark, shake well, measure the amplitude value at 231nm of the third-order derivative spectrum according to the law, and substitute it into the standard curve to calculate the content. The results are shown in the table 1:

Table 1 econazole nitrate recovery measurement result (n=3)

3.5 Sample Determination

Prepare three parts of different lot numbers of Funing emulsifiable cream, the finished product of each lot number detects content by the method under the recovery rate item, and the results are shown in Table 2:

Table 2 Determination results of econazole nitrate in Funing cream (n=2)

4. Stability experiment

Choose three temperatures of 45, 50, and 55°C, respectively, and seal and bake them in an oven for 10 hours. It is found that the appearance does not change only at 45°C, so 45°C is selected as the measurement temperature.

Bake in an oven at 45°C for 0, 4, 8, 12, and 16 hours respectively, record the data, and measure the content, which are 100%, 98%, 95%, 93%, and 93% respectively. The stability is not good, and it cannot last long in summer place.

4.1 Bake continuously in an oven at 55°C for 6 hours, and place in a refrigerator at 0°C for 1 week without oil-water separation.

4.2 Centrifuge at a high speed of 2500 rpm for half an hour, the cream is not separated, but the particles are slightly rough.

Experiments have shown that with a certain amount of Funing cream, increasing, decreasing or increasing the content of matrix and changing the amount of gentamicin sulfate and dexamethasone sodium phosphate, the peak value is basically unchanged, indicating that this method can eliminate interference. The extract is relatively stable. After standing for 48 hours, the absorption value is measured at the measuring wavelength, and the absorption value has no obvious change. The Funing cream to be tested can meet the requirements of hospital preparations.

Claims (4)

1. a quality detection method of Funing emulsifiable paste, comprising the identification of antifungal active ingredient econazole nitrate and the mensuration of its content, it is characterized in that: the identification of described econazole nitrate adopts thin-layer chromatography identification method, Specifically, take 2g of Funing cream to be tested, 20ml of methanol, heat and stir in a water bath at 80°C, cool in an ice bath, filter, evaporate the filtrate to dryness, add 2ml of methanol to dissolve it as the test solution, and accurately weigh 20mg of econazole nitrate sample, 2ml Dissolve in methanol as a control solution, take 5ul points on a silica gel plate, develop with ether, and develop color with iodine vapor for comparison; The determination of the econazole nitrate content is to adopt the third-order reciprocal spectroscopic method, first separate and extract the econazole nitrate in the Funing cream, accurately weigh about 1g of the Funing cream to be tested, add 15ml of solvent, and The solvent is mixed with 0.1mol/L HCL and methanol at a volume ratio of 1:9, heated and stirred in a water bath at 80°C until clarified, cooled in a refrigerator to below room temperature, filtered, washed three times, each time with 10ml of the solvent, Merge the filtrate into a 50ml volumetric flask, add the solvent to dilute to the mark, shake up, accurately measure 2.5ml of the diluent in a 25ml volumetric flask, add the solvent to the mark, and shake up; secondly, use a UV-visible spectrophotometer , Determination of econazole nitrate content at 231nm by third order derivative method. 2. the quality detection method of Funing emulsifiable cream as claimed in claim 1, it is characterized in that: described quality detection method also comprises the identification of antibacterial active ingredient gentamycin sulfate, is specifically to take the Funing emulsifiable cream 0.5 Add 10ml of water to dissolve g, break the emulsion in a boiling water bath, add 0.1wt% (mass percentage concentration) ninhydrin water-saturated n-butanol solution 1ml and pyridine 0.5ml while hot, heat in a water bath at 100°C for 5min, and develop color. 3. the quality detection method of Funing emulsifiable cream as claimed in claim 1 or 2, it is characterized in that: described quality detection method also comprises that dexamethasone sodium phosphate is differentiated, specifically get to-be-tested Funing emulsifiable paste 10g ( Containing 6 mg of dexamethasone sodium phosphate), add 10 ml of methanol, dissolve in 80°C water bath, filter while hot, evaporate the filtrate to dryness with a rotary evaporator after cooling, add the residue to 1 ml of sulfuric acid for color development. 4. A quality detection method for Funing cream, which is the identification of the antifungal active ingredient econazole nitrate and the determination of its content, the identification of the antibacterial active ingredient gentamicin sulfate, and the determination of the dexamethasone sodium phosphate component Carry out identification; It is characterized in that: the identification of described econazole nitrate adopts thin-layer chromatography identification method, specifically take 2g of Funing cream to be tested, 20ml of methanol, heat and stir in a water bath at 80°C, cool in an ice bath, filter, and filtrate Evaporate to dryness, add 2ml methanol to dissolve as test solution, accurately weigh 20mg econazole nitrate sample, 2ml methanol dissolve as contrast solution, divide and get 5ul point on silica gel plate, diethyl ether develops, iodine vapor develops color, compares; The determination of the econazole nitrate content is to adopt the third-order reciprocal spectroscopic method, first separate and extract the econazole nitrate in the Funing cream, accurately weigh about 1g of the Funing cream to be tested, add 15ml of solvent, and the solvent is obtained by Mix 0.1mol/L HCL and methanol at a volume ratio of 1:9, heat and stir in a water bath at 80°C until clarification, put in a refrigerator to cool below room temperature, filter, wash three times, use 10ml of the solvent each time, and combine the filtrates In the 50ml volumetric flask, add the solvent to dilute to the scale, shake up, accurately measure 2.5ml of the diluent in a 25ml volumetric flask, add the solvent to the scale, and shake up; The content of econazole nitrate was determined by the third derivative method at 231nm; To identify gentamicin sulfate, specifically, take 0.5g of Funing cream to be tested (containing 5mg of gentamicin sulfate) and dissolve it in 10ml of water, break the emulsion in a boiling water bath, and add 0.1wt% (mass percentage concentration) indene while it is hot 1ml of water-saturated n-butanol solution of triketone and 0.5ml of pyridine were heated in a water bath at 100°C for 5min to develop color; Dexamethasone sodium phosphate was identified by taking 10g of Funing cream to be tested (containing 6mg of dexamethasone sodium phosphate), adding 10ml of methanol, dissolving in 80°C water bath, filtering while hot, and evaporating to dryness with a rotary evaporator after the filtrate was cooled. , the residue was added to 1ml sulfuric acid for color development.

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