CN101750457B - Method for measuring ciclopirox olamine - Google Patents

Abstract

The invention discloses a method for measuring ciclopirox olamine. A chromatographic column with C18 hydrophilic filler is selected, the clathration of beta-cyclodextrin on ciclopirox olamine is utilized, blended solution of aqueous solution of beta-cyclodextrin and acetonitrile is used as a mobile phase, an ultraviolet detector is adopted, and the method can be used for quantitatively measuring active ingredient ciclopirox olamine in a ciclopirox olamine emulsifiable paste. The method has low detection limit and wide linear range; and the linear range well covers the medicament concentration range of the ciclopirox olamine, so that the method can be well used for fast detection of the ciclopirox olamine in the medicament production process and clinical medicament monitoring.

Description

A kind of method of measuring Ciclopirox Olamine

Technical field

The invention belongs to the analytical chemistry field, relate to a kind of method of measuring Ciclopirox Olamine, a kind of efficient liquid-phase chromatography method of measuring Ciclopirox Olamine in the Ciclopirox Olamine emulsifiable paste through cyclodextrin encapsulated Ciclopirox Olamine of saying so more specifically.

Background technology

Ciclopirox Olamine has another name called ciclopirox olamine, cyclopirox amine.Molecular formula is C ₁₄H ₂₄N ₂O ₃, its structural formula is following:

Molecular weight is 268.36, has the characteristics of the serum infiltration of hypotoxicity and brute force, and it has powerful sterilization functions to dermatophyte, Candida albicans etc., on the fungus-caused skin keratin disease of treatment, effect is preferably arranged.Its concentration can suppress dermatophyte and saccharomycete when being 4～8 μ g/ml; Also there is inhibiting effect (to draw 28-NF23, United States Pharmacopoeia Convention, Inc. to Escherichia coli, proteus, staphylococcus aureus, hemolytic streptococcus etc. during higher concentration (16～78 μ g/ml) from document 1:United States Pharmacopoeia; Rockville; MD, 2005, p.470.).Hence one can see that, and the concentration of Ciclopirox Olamine is very important for the superficial fungi and the candida albicans infection that suppress epidermis or mucous membrane.The method of detection by quantitative chain rate ketoamine concentration has following several kinds at present:

Though being the most frequently used detection technique, the spectrophotometry Ciclopirox Olamine (draws from document 2: Ai Yusuo the spectrophotometry of Ciclopirox Olamine emulsifiable paste content; Northwest pharmaceutical journal .19905 (1) 2～3), but since its accurate quantification and in similar compound the deficiency of specific assay Ciclopirox Olamine, this method can not be applied to the quality control in stability study and the medicine production process.Other has document (to draw from document 3:Escarrone; A.L.V.et al LC-UV method with pre-columnderivatization for the determination of ciclopirox olamine in raw material andtopical solution Chromatographia 200867 (11-12) 967-971) introduces; Use the way detection by quantitative Ciclopirox Olamine of deriving before the performance liquid chromatographic column; Must be through complicated column front derivation step; But the derivatization process makes other component unstable, and is not suitable for the fast detecting in the medicine production process.

American Pharmacopeia (USP) has a kind of method with high performance liquid chromatography detection Ciclopirox Olamine (to draw the 28-NF23 from document 1:United States Pharmacopoeia, United States PharmacopoeiaConvention, Inc.; Rockville, MD, 2005; P.470.); Under its condition, because the strong chelating effect of primary hydroxyl group and ionizable metal salt, there is serious conditions of streaking at the material peak of Ciclopirox Olamine.In order to reduce chelating effect, before experiment, first water: acetonitrile: glacial acetic acid: (500: 500: 1: mixing eluent 1), the flushing chromatographic column is 15h at least, washes 5h at least with moving phase again for diacetone.But, even passed through flushing so for a long time, the detection Ciclopirox Olamine that this method still can not be successful.

Summary of the invention

On the basis of above-mentioned previous work, the object of the present invention is to provide a kind of method of measuring Ciclopirox Olamine.This method is utilized the clathration of beta-schardinger dextrin-, beta-schardinger dextrin-is added in the moving phase of high performance liquid chromatography, makes itself and Ciclopirox Olamine form special inclusion compound, reduces its suction-operated on chromatographic column, utilizes UV-detector to detect Ciclopirox Olamine.

The objective of the invention is to realize through following technical scheme:

A kind of method of measuring Ciclopirox Olamine the steps include:

A. the preparation of Ciclopirox Olamine standard solution

Prepare Ciclopirox Olamine standard items stock solution with acetonitrile as solvent: pipette the stock solution of certain volume,, then above-mentioned standard solution is placed irradiation under the uviol lamp, make standard solution with the solution of dilution in acetonitrile to some parts of variable concentrations;

B. The pretreatment

Accurately take by weighing an amount of Ciclopirox Olamine emulsifiable paste, place the 10mL volumetric flask, add absolute ethyl alcohol 7mL; Use the absolute ethyl alcohol constant volume behind the ultrasonic 20min, continue ultrasonic 5min, with the organic filtering with microporous membrane of 0.45 μ m; Pipetting 1.0mL filtrates in the 10mL volumetric flask; Shake up with the acetonitrile constant volume, then above-mentioned standard solution is placed irradiation under the uviol lamp, sample introduction is preceding with 0.45 μ m organic membrane filter;

C. the mensuration of liquid chromatography

Adopting the chromatographic column of C18 hydrophilic filler, is that water mixes as moving phase with the organic phase acetonitrile with the certain beta-schardinger dextrin-WS of pH, and flow rate of mobile phase is 1mL/min, and column temperature is 30～50 ℃, and the ultraviolet detection wavelength is 298 ± 5nm; Sample size is 20 μ L; Difference sample introduction standard solution and sample solution; Adopt peak area to concentration drawing curve; Obtaining equation of linear regression is: y=ax+b, and utilize sample solution peak area A to combine above-mentioned working curve to calculate the content of Ciclopirox Olamine in the Ciclopirox Olamine Ointment in Treatment, computing formula is following:

Sample solution concentration: $c=\frac{A-b}{a}\left(\mathrm{Ppm}\right)---\left(1\right)$

(1) in the formula: A is the sample solution peak area; A, b are respectively that equation of linear regression is: slope among the y=ax+b and intercept;

The percentage composition of Ciclopirox Olamine in the Ciclopirox Olamine Ointment in Treatment: $w=\frac{100c}{M}\×100\%---\left(2\right)$

(2) c is a sample solution concentration in the formula; M is a Ciclopirox Olamine emulsifiable paste quality;

Advantage of the present invention: this is invented as a kind of mensuration Ciclopirox Olamine method; Beta-schardinger dextrin-is added in the moving phase; Utilize the clathration of beta-schardinger dextrin-to Ciclopirox Olamine; Having overcome Ciclopirox Olamine strong adsorption on chromatographic column can not be caused the shortcoming that it can't quantitative measurement by wash-out, thereby finds a kind of method of effective mensuration Ciclopirox Olamine.This invention has detectability low (detectability can reach 0.01ppm); The range of linearity is wide by (0.31～75.00ppm); The recovery is 95.9%; RSD=4.38%, its range of linearity has well covered the medication concentration range (4～8 μ g/ml) of Ciclopirox Olamine, therefore can well be used for the fast detecting and the clinical application monitoring of medicine production process Ciclopirox Olamine.

Description of drawings

Fig. 1 is a beta-cyclodextrin inclusion compound Ciclopirox Olamine structural representation.

Fig. 2 is the pure article standard of a Ciclopirox Olamine chromatogram.

Fig. 3 is the Ciclopirox Olamine standard working curve.

Fig. 4 is a Ciclopirox Olamine emulsifiable paste actual sample chromatogram, and 1 is the peak of sample 1, and 2 is the peak of sample 2.

The practical implementation method

The present invention adopts the Varian high performance liquid chromatography (to comprise ProStar 210 pumps; The 7725i sampling valve is 20 μ L injection annulus; UV-detector is ProStar325, and the chromatographic data analytic system is StarWS) and AE.LICHROM C18 5 μ m 150 * 4.6mm chromatographic column;

A kind of method of measuring Ciclopirox Olamine, its implementation process are divided into following four steps:

A. the preparation of Ciclopirox Olamine standard solution

Get 5.0mg Ciclopirox Olamine standard items, in the 5mL volumetric flask, make the stock solution of 1mg/mL with the acetonitrile constant volume; Pipetting volume respectively is the stock solution of 1.55 μ L, 3.15 μ L, 6.25 μ L, 12.50 μ L, 25.00 μ L, 50.00 μ L, 100.00 μ L, 250.005 μ L, 375.00 μ L, is respectively the solution of 0.31ppm, 0.63ppm, 1.25ppm, 2.50ppm, 5.00ppm, 10.00ppm, 20.00ppm, 50.00ppm, 75.00ppm with dilution in acetonitrile to concentration.Then above-mentioned standard solution is placed under the uviol lamp and shone 24 hours, make standard solution.

B. The pretreatment

Accurately take by weighing Ciclopirox Olamine emulsifiable paste 0.10g, 0.20g (production of field pharmaceutcal corporation, Ltd of Hubei section), be labeled as sample 1, sample 2 respectively; Place the 10mL volumetric flask then respectively, add absolute ethyl alcohol 7mL, use the absolute ethyl alcohol constant volume behind the ultrasonic 20min; Continue ultrasonic 5min; With the organic filtering with microporous membrane of 0.45 μ m, pipette 1.0mL and filtrate in 10mL volumetric flask for use, with the acetonitrile constant volume and shake up; Then above-mentioned constant volume solution is placed under the uviol lamp irradiation 24 hours, before the sample introduction with solution with 0.45 μ m organic membrane filter.

C. the drafting of working curve

The drafting of working curve of the present invention; Adopt the chromatographic column of C18 hydrophilic filler, with the beta-schardinger dextrin-WS of the 13.2mmol/L of pH=4.0: acetonitrile=50: 50 (V/V) is a moving phase, and flow rate of mobile phase is 1mL/min; Column temperature is 40 ℃, and the ultraviolet detection wavelength is 298nm; The finite concentration standard Ciclopirox Olamine standard solution 20 μ L that prepare in the sample introduction a step successively; Beta-schardinger dextrin-and Ciclopirox Olamine form inclusion compound; By the moving phase wash-out, adopt peak area to concentration drawing curve then, equation of linear regression is: y=9.6519x+2.3721; The range of linearity is 0.31～75.00ppm, linearly dependent coefficient R ²=0.9994, working curve is seen Fig. 3.

D. the mensuration of sample

Get the Ciclopirox Olamine emulsifiable paste sample feeding that 20 μ L handle well, under above-mentioned c step condition, measure.According to formula (1), formula (2), combine above-mentioned working curve to calculate the percentage composition of Ciclopirox Olamine in the Ciclopirox Olamine Ointment in Treatment with peak area.

Figure can know by Fig. 4 sample chromatogram, and the ratio of sample peak 1 and the area at sample peak 2 equals the mass ratio of sample 1 and sample 2 in the b step in the spectrogram, and visible in the range of linearity of 0.31～75.00ppm, the quantitative measurement of Ciclopirox Olamine is accurately; And then the Mass Calculation that combines sample 1 and sample 2 goes out that the percentage composition of Ciclopirox Olamine is 1% in the Ciclopirox Olamine Ointment in Treatment, is consistent with the product description of this ointment.

Claims (1)

1. a method of measuring Ciclopirox Olamine the steps include:

A. the preparation of Ciclopirox Olamine standard solution

Prepare Ciclopirox Olamine standard items stock solution with acetonitrile as solvent: pipette the stock solution of certain volume, with the solution of dilution in acetonitrile to some parts of variable concentrations, the solution with variable concentrations places irradiation under the uviol lamp then, makes standard solution;

B. The pretreatment

Accurately take by weighing an amount of Ciclopirox Olamine emulsifiable paste, place the 10mL volumetric flask, add absolute ethyl alcohol 7mL; Use the absolute ethyl alcohol constant volume behind the ultrasonic 20min, continue ultrasonic 5min, with the organic filtering with microporous membrane of 0.45 μ m; Pipetting 1.0mL filtrates in the 10mL volumetric flask; Shake up with the acetonitrile constant volume, then above-mentioned constant volume solution is placed irradiation under the uviol lamp, sample introduction is preceding with 0.45 μ m organic membrane filter;

C. the mensuration of liquid chromatography

Adopt the chromatographic column of C18 hydrophilic filler, with the beta-schardinger dextrin-WS of the 13.2mmol/L of pH=4.0: acetonitrile=50: 50V/V is a moving phase, and flow rate of mobile phase is 1mL/min, and column temperature is 40 ℃, and the ultraviolet detection wavelength is 298nm; Sample size is 20 μ L; Difference sample introduction standard solution and constant volume solution; Adopt peak area to concentration drawing curve; Obtaining equation of linear regression is: y=ax+b, and utilize sample solution peak area A to combine above-mentioned working curve to calculate the percentage composition of Ciclopirox Olamine in the Ciclopirox Olamine Ointment in Treatment, computing formula is following:

Sample solution concentration: $c=\frac{A-b}{a}---\left(1\right)$

(1) in the formula: A is the sample solution peak area; A, b are respectively that equation of linear regression is: slope among the y=ax+b and intercept; The sample solution concentration unit is ppm;

The percentage composition of Ciclopirox Olamine in the Ciclopirox Olamine Ointment in Treatment: $w=\frac{100c}{M}\×100\%---\left(2\right)$

(2) c is a sample solution concentration in the formula; M is a Ciclopirox Olamine emulsifiable paste quality.

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