CN107505424A - The HPLC assay methods of PVP content in the PVP-I aqueous solution - Google Patents

Abstract

The invention belongs to the field of analytical chemistry, and relates to a method for determining the content of povidone in a povidone-iodine solution, in particular to an HPLC method for determining the content of povidone in an aqueous solution of povidone-iodine. Adopt C18 chromatographic column in this method, take acetonitrile-water 5: 95 as mobile phase, with the flow rate of ultraviolet detector 0.8mL/min, the ultraviolet detection wavelength of 205nm measures povidone content in povidone-iodine aqueous solution, through detection The analysis of the sample is determined and the results are displayed. The HPLC assay method of povidone-iodine content in the povidone-iodine aqueous solution of the present invention is simple, quick, good in accuracy, high in sensitivity, is especially suitable for measuring the content of povidone in the povidone-iodine aqueous solution, and the inventive method helps Improve the quality control and drug safety of povidone-iodine liquid preparations.

Description

The HPLC determination method of povidone content in povidone-iodine aqueous solution

technical field

The invention belongs to the field of analytical chemistry, and relates to a method for determining the content of povidone in a povidone-iodine solution, in particular to an HPLC method for determining the content of povidone in an aqueous solution of povidone-iodine.

Background technique

The prior art discloses that povidone-iodine (PVP-I) is a compound of 1-vinyl-2-pyrrolidone polymer and iodine, and has been made into various dosage forms such as ointment, gel, suppository and liquid preparation , Widely used in disinfection and infection of skin and mucous membranes. Application practice shows that povidone-iodine solution, as a disinfectant, can slowly release iodine to exert antibacterial effect, and has the advantages of less tissue irritation, relatively stable performance, and less toxicity. In 2014, the Ministry of Agriculture of the People's Republic of China officially approved povidone-iodine oral liquid to be launched as a new drug for the treatment of diarrhea in pigs, chickens and other livestock. After testing by many companies, the use effect is good. But in practice at present, there are such as, the content of povidone in the povidone-iodine liquid preparation is different, and the degree of complexation with iodine is different, causing many solutions to be unstable and prone to defects such as photolysis. Therefore, the determination of povidone-iodine content in povidone-iodine solution is particularly important for the quality control and drug safety of povidone-iodine preparations, and povidone is an important pharmaceutical excipient in the world, so far There is no report on the determination method.

Based on the status quo of the prior art, the inventor of the present application intends to provide a method for determining the content of povidone in povidone-iodine solution, especially an HPLC method for determining the content of povidone in povidone-iodine aqueous solution.

Contents of the invention

The object of the invention is to overcome the defective of prior art, provide the assay method of povidone content in povidone iodine solution, relate in particular to a kind of high performance liquid chromatography ( HPLC).

For this reason, the technical scheme that the present invention adopts is:

Adopt C18 chromatographic column, take acetonitrile-water mixed solvent as mobile phase, utilize ultraviolet detector to measure povidone content in povidone-iodine aqueous solution;

In the present invention, the volume ratio of the acetonitrile-water mixed solvent is 5:95, the retention time of the povidone chromatographic peak is 4.60min, and the peak shape is better;

In the present invention, the flow rate is 0.8mL/min, the ultraviolet detection wavelength is 205nm, and the povidone chromatographic peak is not interfered by the solvent peak and other components in the sample;

The method of the present invention is suitable for detecting the povidone-iodine content in the povidone-iodine liquid preparation. In one embodiment of the present invention, the povidone-iodine aqueous solution is used as the sample to be tested.

In the HPLC assay method of povidone content in povidone-iodine aqueous solution provided by the invention, reference substance solution and sample solution are prepared by following steps:

1) Take about 0.01g of the povidone reference substance, weigh it accurately, put it in a 10mL volumetric flask, add water to dissolve, set the volume to the mark, and shake well to obtain a 1mg/mL povidone reference substance solution;

2) Take about 2mL of povidone-iodine aqueous solution, measure it precisely, put it in a 10mL volumetric flask, add water to dilute, set the volume to the mark, and shake well to obtain a solution with a concentration of 10mg/mL. Measure 1mL of this solution precisely and place it in a 10mL volumetric flask , diluted with water to the mark, and shaken to obtain a povidone-iodine sample solution.

Through the analysis and determination of the test samples, the results show that. The HPLC assay method of povidone-iodine content in the povidone-iodine aqueous solution of the present invention is simple, quick, good in accuracy, high in sensitivity, is especially suitable for measuring the content of povidone in the povidone-iodine aqueous solution, and the inventive method helps Improve the quality control and drug safety of povidone-iodine liquid preparations.

Description of drawings

Figure 1 is a blank solvent (water) high performance liquid chromatogram.

Fig. 2 is the high performance liquid chromatogram of povidone reference substance solution,

Wherein: 1 is povidone.

Figure 3 is a high performance liquid chromatogram of povidone-iodine aqueous solution dilution liquid,

Wherein: 1 is povidone.

Fig. 4 is the linear graph of the peak area of the povidone reference substance to the concentration,

Wherein: abscissa is povidone reference substance concentration (mg/ml), and ordinate is povidone peak area.

Specific embodiments of the present invention are described in detail below. The following examples are only to illustrate the technical ideas of the present invention, and cannot limit the protection scope of the present invention with this. Any changes made fall within the protection scope of the present invention.

detailed description:

Example 1

1. Instruments and reagents

1.1 Instrument

Shimadzu LC-10AD high performance liquid chromatograph, including LC-10AD quaternary pump, SPD-A ultraviolet detector, STO-10A column thermostat, manual injector with 20 μ L quantitative loop (Shimadzu Corporation of Japan); HS2000 Chromatography data workstation (Hangzhou Yingpu Technology Development Co., Ltd.);

1.2 Reagent

Povidone reference substance (RE-GOLD CHEMICAL CO.LTD, purity 99.8% (calculated as dry product), batch number: 03700184807); povidone-iodine aqueous solution (made in the laboratory); acetonitrile (HPLC pure, J&KCHEMICA, batch number: LP50K04 ); deionized water (made by Millipore Direct-Q pure water generator);

2. Method

2.1 Preparation of solution

Preparation of Povidone Reference Substance Solution: Take about 0.01g of Povidone Reference Substance, accurately weighed, put in a 10mL volumetric flask, add water to dissolve, set the volume to the mark, and shake well to obtain 1mg/mL Povidone Reference Substance Solution ;

Preparation of povidone-iodine sample solution: Take about 2mL of povidone-iodine aqueous solution, measure it precisely, put it in a 10mL volumetric flask, add water to dilute, set the volume to the mark, shake well, and obtain a solution with a concentration of 10mg/mL; measure it precisely Put 1mL of this solution in a 10mL volumetric flask, dilute to the mark with water, and shake well to obtain povidone-iodine sample solution;

2.2 Chromatographic conditions

Column: Dikma C18 column (4.6mm×150mm, 5μm)

Mobile phase: acetonitrile-water volume ratio of 5:95

Flow rate: 0.8mL/min

UV detection wavelength: 205nm

Column temperature: 20°C

Injection volume: 20μL

2.3 Methodological investigation

2.3.1 Chromatographic system applicability test: take 20 μL of povidone reference substance solution with a concentration of 1 mg/mL for sample analysis, and the number of theoretical plates is not less than 2500 based on the povidone peak;

2.3.2 method specificity: measure reference substance, sample solution according to this method, obtain corresponding retention time t=4.60min, povidone chromatographic peak is not interfered by solvent peak and other components in the sample, and peak shape is better ( As shown in Figures 1, 2, and 3);

2.3.3 Linear range: prepare reference substance solutions with concentrations of 0.1, 0.2, 0.4, 0.6, 0.8, and 1 mg/mL, take 20 μL of each sample, analyze according to the above chromatographic conditions, and use povidone peak area A (n= 6) Plot the mass concentration C (mg/mL) (the standard curve is shown in Figure 4), and obtain the linear regression equation: A=554347C+5484.6 (r=0.9998, n=6), and the results show that: in the range of 0.1 to 1 mg/mL Within the scope of , povidone peak area A and concentration C are in good linear relationship;

2.3.4 Method sensitivity: the reference substance solution with the minimum concentration in the linear range was gradually diluted with water, and after determination, the lowest detection limit of povidone obtained by this method was 0.05 μg/mL (S/N=3); the limit of quantification was 0.2 μg/mL (S/N=10);

2.3.5 Method accuracy (indicated by recovery rate): prepare reference substance solutions with concentrations of 0.3, 0.5 and 0.7 mg/mL, and perform recovery determination according to the law. The recovery rate of the reference substance solution is shown in Table 1.

Table 1 HPLC method measures povidone content method recovery rate (%, n=3)

2.3.6 Method precision: prepare sample solutions with concentrations of 0.2, 0.6 and 1 mg/mL, analyze according to the above chromatographic conditions, measure three times in one day, and measure intraday precision; measure continuously for three days, measure interday precision , the results are shown in Table 2;

Table 2 HPLC method measures the precision experiment result of povidone reference substance content

2.3.7 Method stability: Prepare a sample solution with a concentration of 1mg/mL, take 20μL samples at 0, 1, 3, 5, 7, 9, 12, and 24 hours respectively, analyze according to the above chromatographic conditions, and investigate the stability of the solution. Stability; the results showed that there was no significant change in the drug peak area within 24h, and the peak area RSD=0.02%, indicating that the sample was determined to be stable within 24h;

2.3.8 Sample recovery rate: Accurately measure three groups (three copies each) of povidone-iodine sample solution (about 0.25 mg of povidone), put it in a volumetric flask, add concentration of 0.20 mg/ mL, 0.25mg/mL, 5mL of 0.30mg/mL povidone standard solution (equivalent to 80%, 100%, 120% of the povidone content in the sample solution, that is, 0.20mg, 0.25mg, 0.30mg of povidone Vitone), respectively get three parts of the solution after sample addition and carry out HPLC method determination, each injection 20uL, the result is as shown in table 3; The calculation formula of sample addition recovery is:

Sample addition recovery rate=(actually measured concentration-1/2 sample concentration)/(1/2 added standard concentration)×100%.

The sample addition recovery measurement result (%, n=3) of table 3 povidone

2.4 Determination of povidone content in samples:

Get 20 μ L of povidone-iodine sample solution for sample analysis, and record the chromatogram; the calculation method of the determination result of the sample content is: the povidone peak area in the sample solution measured is brought into the standard curve, and the povidone in the sample solution is obtained. content, that is

The percentage content of povidone=0.1 * 50 * the content of povidone in the sample solution in povidone-iodine aqueous solution

The calculation results are shown in Table 4:

Table 4HPLC method measures the content result of povidone in povidone-iodine aqueous solution

The experimental results of the present invention show that the contents of povidone in the three povidone-iodine aqueous solutions measured by the HPLC method are 1.21%, 1.16%, and 1.19%, respectively.

Claims (6)

1. the HPLC assay method of povidone-iodine content in povidone-iodine aqueous solution, it is characterized in that, adopt C18 chromatographic column in this method, take acetonitrile-water mixed solution as mobile phase, utilize ultraviolet detector to measure povidone-iodine aqueous solution Povidone content. 2. the HPLC assay method of povidone content in povidone-iodine aqueous solution according to claim 1, is characterized in that: the volume ratio of described acetonitrile-water mixed solution is 5: 95. 3. the HPLC assay method of povidone content in povidone-iodine aqueous solution according to claim 1, is characterized in that: in described method, the flow velocity that adopts is 0.8mL/min. 4. the HPLC assay method of povidone-iodine content in the povidone-iodine aqueous solution according to claim 1, is characterized in that: in described method, the ultraviolet detection wavelength that adopts is 205nm. 5. the HPLC assay method of povidone content in povidone-iodine aqueous solution according to claim 1, is characterized in that: prepare reference substance solution and sample solution by following steps in the described method: 1) Get the povidone reference substance, accurately weigh it, put it in a volumetric flask, add water to dissolve, set the volume to the mark, and shake up to obtain a 1mg/mL povidone reference substance solution; 2) Take povidone-iodine solution, measure it precisely, put it in a volumetric flask, add water to dilute, set the volume to the mark, and shake well to obtain a solution with a concentration of 10mg/mL. Measure 1mL of this solution precisely and place it in a 10mL volumetric flask, add water Dilute to the mark and shake well to obtain povidone-iodine sample solution. 6. the HPLC assay method of povidone-iodine content in the povidone-iodine aqueous solution according to claim 1 or 5, is characterized in that: in described method, adopts povidone-iodine aqueous solution to be detected sample.