CN111103370B - Method for simultaneously separating and quantifying multiple components - Google Patents

Abstract

The invention provides a method for simultaneously separating and quantifying a plurality of components, which comprises the steps of sample pretreatment and sample detection. The method has better specificity, and can avoid the interference of various components, especially the components (such as polyethylene glycol and povidone) which are easy to interfere with each other to the measurement. The invention provides a good method for reverse engineering, component analysis, auxiliary material release detection and the like of pharmaceutical preparations and other products (such as cosmetics and foods). The method is simple to operate, has good specificity and accuracy, wide linear range and high sensitivity, and greatly improves the detection capability and the detection efficiency.

Description

Method for simultaneously separating and quantifying multiple components

Technical Field

The invention belongs to the technical field of analysis, mainly relates to a method for simultaneously separating and quantifying multiple components, and more particularly relates to a method for simultaneously separating and quantifying acrylic resin, povidone and polyethylene glycol in multiple components.

Background

Acrylic resins (Polymethacrylates) available under the tradename Eudragit (Eudragit) include methacrylic acid copolymers and methacrylate ester copolymers. The Eudragit is widely used for gastric coating, enteric coating, sustained-release coating, protective isolation coating, sustained-release matrix material of pharmaceutical preparation and matrix adhesive material of transdermal drug delivery preparation. Since the first product appeared for half a century now, the product has been widely used in various pharmaceutical preparations and becomes an important auxiliary material for many international famous brand preparation products. The acrylic resin detection methods reported so far are mainly titration method [ Methacrylic Acid and Ethyl Acrylate Copolymer [ S ]. USP42-NF37,2018:5823] and gel chromatography method [ Fructus pericarpus, Zhao Li, Cao Sha, determination of molecular weight and distribution of acrylic resin [ J ]. Guangzhou chemical industry, 2014,42(24):103-104 ].

Povidone (Povidone) has wide application in medicine, is mainly applied to solid preparations, and is most widely applied to binding agents of tablets and granules. Povidone can also be used as a disintegrant, solubilizer, coating material, suspending agent, stabilizer, or viscosity enhancer. Currently, the method for detecting povidone has been reported mainly to be nitrogen determination [ povidone K30[ S ]. four parts of Chinese pharmacopoeia 2015 edition, 2015:631, etc. ] and high performance liquid chromatography [ Longjiakun, Jiangjie ice, Zhao Meiyan, etc. ] using reversed phase silica gel C18 as stationary phase and HPLC method to determine the content of povidone in povidone iodine oral liquid [ J ]. Chinese clinical medicine journal, 2016,25(5):300-

Polyethylene Glycol (PEG) is used in the pharmaceutical industry as an excipient for the preparation of suppositories, ointments. Polyethylene glycols can also be used as suspending agents, plasticizers, lubricants, and the like. It is suitable for various industries such as medicine, fertilizer, paper making, ceramics, detergent, cosmetics, heat treatment, water treatment, fire fighting, oil exploitation, etc. It has been reported that Polyethylene Glycol detection methods mainly include gel chromatography [ Polyethylene Glycol 3350[ S ]. USP42-NF37,2017:3557, etc. ], hydrophobic interaction chromatography [ James Ervin Seely, Louisville, Colo. Using. of hydrophilic interaction chromatography to purify Polyethylene Glycol (P). US005747639A,1998-05-05], high performance liquid chromatography using C18, C8, C4 reverse phase silica gel as a stationary phase [ high performance liquid chromatography for measuring Polyethylene Glycol content in polyethoxylated nonionic surfactants (requisition draft) [ J ]. daily chemical science, 2015,38(6):50-52, etc. ], spectrophotometry [ Polyethylene Glycol residue measurement method [ four parts of Chinese pharmacopoeia 2015 edition, 2015:238, etc. ].

At present, no literature report method is available for simultaneous separation and quantitative detection of various components such as acrylic resin, povidone, polyethylene glycol and the like. Reported detection methods (gel permeation chromatography, titration method, nitrogen determination method, reversed phase silica gel or hydrophobic interaction chromatography, spectrophotometry and the like) of acrylic resin, povidone and polyethylene glycol are poor in specificity, other components are easy to interfere with determination, and multiple components cannot be separated and quantified simultaneously. Measuring the content of acrylic resin: by adopting an acid-base titration principle, a plurality of acidic components are easy to interfere with the method determination; the reported liquid gel chromatography is adopted, and components such as polyethylene glycol are easy to interfere the determination of the acrylic resin, and the simultaneous separation and quantification of various components cannot be realized. Measuring the content of the polyvidone: when the content of the povidone is detected by a nitrogen determination method, a plurality of components in a sample contain nitrogen elements, so that interference is easy to generate; the high performance liquid chromatography adopting the reversed phase silica gel C18 reported in the literature as the stationary phase can only realize the content determination of the povidone and cannot realize the simultaneous separation and quantitative detection of various components when an ultraviolet detector is used for detection. Measuring the content of polyethylene glycol: by adopting the reported liquid gel chromatography, the components such as the povidone and the like are easily overlapped with the chromatographic peak of the polyethylene glycol and cannot be separated and quantified; by adopting a hydrophobic interaction chromatography or a high performance liquid chromatography which adopts reversed phase silica gel C18, C8 and C4 as a stationary phase, because a plurality of components in a sample are strong polar compounds, the peaks appear before and after a solvent peak, and the simultaneous separation of the plurality of components is difficult to realize; the spectrophotometry is adopted, the operation is troublesome (derivatization is needed), the interference of other components is easy, the specificity is poor, the linear range is narrow, and the simultaneous separation and quantification of various components cannot be realized.

Meanwhile, in the existing gel permeation chromatography, when the molecular weight of components such as acrylic resin is detected, if strong polar reagents such as N, N-dimethylformamide are used as a mobile phase, salts such as lithium bromide and lithium chloride are often added into the strong polar reagents to eliminate bonding, and the salts such as lithium bromide and lithium chloride have great damage to a chromatographic column.

Aiming at the problems that the detection method has poor specificity and can not simultaneously detect a plurality of components, the invention provides a method for simultaneously separating and quantifying the plurality of components. The method has good specificity, and can separate acrylic resin, polyvidone, polyethylene glycol, etc. and perform accurate quantitative analysis. Provides a good method for reverse engineering, component analysis and auxiliary material release detection of pharmaceutical preparations and other products (such as cosmetics and foods). Meanwhile, the method does not need to add salts such as lithium bromide, lithium chloride and the like into the mobile phase, and the loss of the chromatographic column is obviously reduced.

Disclosure of Invention

The purpose of the invention is as follows: aiming at the problems in the prior art, the invention provides a method for simultaneously separating and quantifying a plurality of components, in particular to a method for simultaneously separating and quantifying acrylic resin, povidone and polyethylene glycol in a plurality of components.

In order to solve the technical problems, the invention adopts the following technical scheme:

a method for simultaneously separating and quantifying a plurality of components, in particular to a method for simultaneously separating and quantifying acrylic resin, povidone and polyethylene glycol in a plurality of components, which comprises the following steps:

(1) pretreating a sample to be detected to prepare a sample solution;

(2) measuring the sample obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result of the sample solution; wherein the chromatographic column is USP liquid chromatographic column classification L21; the mobile phase comprises any one of Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), N-methylpyrrolidone (NMP) and dimethylacetamide (DMAc); the detector is a general purpose detector. Preferably, the quantitative determination method of each component of the sample solution is an external standard method.

The sample is a multi-component mixture, and the mixture comprises but is not limited to any one or a combination of more of acrylic resin, povidone, polyethylene glycol, bulk drug, triethyl citrate, microcrystalline cellulose, sodium lauryl sulfate, talcum powder, magnesium stearate, fumaric acid, crospovidone, saccharin sodium, anhydrous silicon dioxide, magnesium stearate, sodium hydroxide, strawberry-flavor essence, licorice-flavor essence and anhydrous silicon dioxide.

In some embodiments, the acrylic resin types include, but are not limited to, ewing L100-55, ewing L100, ewing L30D-55; povidone models include, but are not limited to, povidone K30(PVP K30), povidone K90(PVP K90), povidone K29/K32(PVP K29/K32), povidone S630(PVP S630); polyethylene glycol types include, but are not limited to, polyethylene glycol 400(PEG 400), polyethylene glycol 600(PEG 600), polyethylene glycol 6000(PEG 6000), and polyethylene glycol 8000(PEG 8000).

Preferably, the components may be of a pharmaceutical adjuvant grade, or of a food or cosmetic grade.

Specifically, the pretreatment step includes: adding a sample to be detected into a volumetric flask, adding a mobile phase, ultrasonically dissolving or uniformly dispersing the sample, and then carrying out constant volume filtration.

Preferably, the ultrasonic time is 5-30 min; after the volume is fixed, the solution is filtered through a polytetrafluoroethylene or equivalent filter membrane with the diameter of 0.22 μm or 0.45 μm.

Preferably, the mobile phase does not contain lithium bromide (LiBr) or lithium chloride (LiCl).

Preferably, after the sample is pretreated, the concentration of the acrylic resin is 0.02 mg/ml-10 mg/ml; the concentration of the povidone is 0.1-10 mg/ml; the concentration of the polyethylene glycol is 0.01-10 mg/ml. The concrete contents of the acrylic resin, the povidone and the polyethylene glycol in the sample have no influence on the determination, and only need to be within the concentration range after treatment.

In the step (2), the flow rate of the gel permeation chromatography is 0.2-2 ml/min, the sample injection amount is 5-100 mu l, and the column temperature is 15-40 ℃. More preferably, the flow rate of gel permeation chromatography is 0.6ml/min, the amount of sample is 20. mu.l, and the column temperature is 35 ℃.

The chromatographic column is an AgilentPLgel series, a Tosoh Bioscience TSKgelHxl and Hhr series, a PhenomenexPhenogel series or a Waters Corp. Styragel HR series in a USP liquid chromatographic column classification L21 series chromatographic column.

In some embodiments, the chromatography column is a single chromatography column; in other embodiments, the chromatography column is a series of multiple chromatography columns. More preferably, the chromatographic column is Agilent PLGel 5 μm MIXED-D, 7.5X 300mm, two or three in series. The chromatographic column uses rigid styrene-divinylbenzene copolymer microspheres (diameter of 3-30 μm) as filler.

Preferably, the universal detector comprises a differential refraction detector (RID), an Evaporative Light Scattering Detector (ELSD).

Has the beneficial effects that: compared with the prior art, the invention has the following advantages:

(1) according to the invention, at least three components (acrylic resin, povidone and polyethylene glycol) are synchronously separated and quantified, so that the detection efficiency is greatly improved;

(2) the invention has better specificity, can avoid the interference of various components, especially the components (such as polyethylene glycol and povidone) which are easy to interfere with each other to the measurement;

(3) the method is simple to operate, wide in linear range, low in quantitative limit and high in accuracy;

(4) according to the method, salts such as lithium bromide and lithium chloride do not need to be added into the mobile phase, and the loss of the chromatographic column is obviously reduced.

Drawings

FIG. 1 is a chromatogram of a control solution of L100-55, PEG6000, PVP K30;

FIG. 2 is a superimposed chromatogram in a specificity test;

FIG. 3 shows the results of L100-55 linear investigation;

FIG. 4 shows the results of a PEG6000 linear investigation;

FIG. 5 shows the results of a linear assay of PVP K30;

FIG. 6 shows the effect of adding different concentrations of lithium bromide to mobile phase DMF on the degree of separation;

FIG. 7 shows the results of chromatography under the chromatographic conditions of examples 8, 9 and 10.

Detailed Description

The invention provides a method for simultaneously separating and quantitatively measuring acrylic resin, povidone and polyethylene glycol in multiple components.

Specifically, the method comprises the following steps:

(1) dissolving a sample to be detected containing acrylic resin, povidone and polyethylene glycol in a mobile phase, wherein the mobile phase comprises any one of dimethylformamide, dimethyl sulfoxide, N-methylpyrrolidone and dimethylacetamide, dissolving the mobile phase by ultrasonic treatment, and filtering to obtain a sample solution;

(2) measuring the sample solution obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result of the sample solution; wherein the chromatographic column is USP liquid chromatographic column classification L21; the detector is a general purpose detector.

The present invention will be described in detail below with reference to specific examples.

Example 1: l100-55, PEG6000 and PVP K30 were separated and analyzed quantitatively.

(1) Taking 15mg of L100-55, PEG6000 and PVP K30 respectively, placing into a 20ml measuring flask, dissolving with DMF, diluting to scale, and shaking to obtain control solution. Taking 2 tablets of a sample to be detected (a roxithromycin dispersible tablet, the specification of which is 50mg, the trade name of which is Rulid, according to the Rulid specification, the roxithromycin dispersible tablet contains auxiliary materials such as microcrystalline cellulose, fumaric acid, methacrylic resin, crospovidone, polyethylene glycol 6000, talcum powder, saccharin sodium, anhydrous silicon dioxide, magnesium stearate, triethyl citrate, sodium dodecyl sulfate, sodium hydroxide, strawberry flavor essence, liquorice flavor essence and the like), adding DMF (dimethyl formamide) to 2/3 volume of a volumetric flask, carrying out ultrasonic treatment for 15min to uniformly disperse the sample (ensuring that L100-55, PEG6000 and PVP K30 in the sample can be completely dissolved), adding DMF to a constant volume to scale, shaking uniformly, and filtering to obtain a sample solution.

(2) Measuring the reference substance solution and the sample solution obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement results;

wherein, the chromatographic conditions are as follows:

the chromatographic columns are Agilent PLgel 5 μm MIXED-D,7.5 × 300mm, and the two columns are connected in series; the mobile phase was DMF, the detector was a differential Refractive Index Detector (RID), the column temperature and detector temperature were 35 deg.C, the flow rate was 0.6ml/min, and the sample loading was 20. mu.l.

The chromatogram of the control solution is shown in FIG. 1, the separation degrees of L100-55 and PEG6000 and the separation degrees of PEG6000 and PVP K30 are respectively 4.6 and 0.7, and the simultaneous separation and quantitative analysis of the three components of L100-55, PEG6000 and PVP K30 can be realized.

The detection result of the sample solution shows that the roxithromycin dispersible tablets Rulid contain L100-5511.9 mg/tablet and PEG 60006.7 mg/tablet, and PVP K30 is not detected in the roxithromycin dispersible tablets.

Example 2: and (3) researching the specificity, linearity, accuracy and quantitative limit of the detection method.

The specificity is as follows:

the mobile phase (DMF) and the special solution (the special solution contains crude drug (roxithromycin), microcrystalline cellulose, fumaric acid, crospovidone, talcum powder, saccharin sodium, anhydrous silicon dioxide, magnesium stearate, triethyl citrate, sodium dodecyl sulfate, sodium hydroxide, strawberry flavor essence and liquorice flavor essence) are subjected to sample injection detection respectively, the experimental result is shown in figure 2, and the result shows that the mobile phase and the special solution have no interference on the determination of L100-55, PEG6000 and PVP K30.

(II) linearity:

preparing series of linear solution injection, performing linear regression with the concentration as abscissa (x) and peak area as ordinate (y) to obtain linear regression equation (shown in FIG. 3-FIG. 5) and square of correlation coefficient (R) of L100-55, PEG6000, PVP K30²) Respectively 1.000, 1.000 and 0.999 (no less than 0.99). I.e., L100-55, PEG6000, PVP K30 concentrations ranged from 0.1mg/ml to 10mg/ml, all showing acceptable linearity.

(III) accuracy:

preparing tablets containing different contents of PEG6000, PVP K30 and L100-55 according to the handbook of pharmaceutical adjuvants (R.C. Luo, P.J, Ski, P.J. Wahler edition, Liujun folk translation): the concentration of polyethylene glycol in the pharmaceutical preparation is 0-30%; the concentration of the povidone in the pharmaceutical preparation is 0-25%; the concentration of the acrylic resin in the medicinal preparation is 0-50%. The tablets contained conventional amounts of the drug substance (roxithromycin), microcrystalline cellulose, fumaric acid, crospovidone, talc, sodium saccharin, anhydrous silicon dioxide, triethyl citrate, and sodium lauryl sulfate in addition to PEG6000, PVP K30, and L100-55, and the contents of PEG6000, PVP K30, and L100-55 added to the tablets were changed to prepare sample solutions, and the recovery rate of the method was confirmed by using this method. The results show that the recovery is between 86 and 108% and the process is accurate. Data results are shown in table 1.

TABLE 1L 100-55, PEG6000, PVP K30 accuracy results

And (IV) determination of a quantitative limit:

preparing sample solutions with different concentrations, recording that when the concentration with the signal-to-noise ratio of about 10 is the limit of quantitation (LOQ) concentration, the peak area RSD of the main peak in the chromatogram of the 6-pin LOQ solution by repeated sample injection is not more than 10%. Data results are shown in table 2.

TABLE 2L 100-55, PEG6000, PVP K30 quantitative limit, detection limit results

The experiment results in: the limit of quantitation of L100-55 is 25.1. mu.g/ml; the limit of quantitation of PEG6000 is 12.5 mu g/ml; the limit of quantitation of PVP K30 was 100.9. mu.g/ml.

Example 3: effect of adding lithium bromide in different concentrations to DMF in Mobile phase on the degree of separation

This example was followed by the same procedure as in example 1 except that the mobile phase and diluent in example 1 were changed from DMF to DMF (containing 0.01% LiBr), DMF (containing 0.05% LiBr), DMF (containing 0.1% LiBr).

The chromatogram is shown in fig. 6, the separation degree data is shown in table 3, if lithium bromide and other salts are added into the mobile phase, the separation degrees among L100-55, PEG6000 and PVP K30 are poor, the three components cannot be completely separated, and the three components of L100-55, PEG6000 and PVP K30 cannot be simultaneously separated and quantified.

TABLE 3 Effect of adding different concentrations of lithium bromide to the mobile phase DMF on the degree of separation

Example 4: detection of different types of acrylic resin, polyethylene glycol and polyvidone

(1) Taking 15mg of L100-55, PEG 400 and PVP K29/K32 respectively, placing in a same 20ml measuring flask, dissolving and diluting to scale with DMF, and shaking up to obtain a reference substance solution 1; taking 15mg of L100, PEG600 and PVP K90 respectively, placing the L100, the PEG600 and the PVP K90 in the same 20ml measuring flask, dissolving the L100, the PEG600 and the PVP K90 in DMF, diluting the solution to a scale, and shaking the solution uniformly to obtain a reference substance solution 2; taking 15mg of L30D-55, PEG8000 and PVP S630 respectively, placing in a same 20ml measuring flask, dissolving with DMF, diluting to scale, and shaking to obtain reference substance solution 3;

(2) measuring the reference substance solution obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result;

wherein, the chromatographic conditions are as follows:

the chromatographic columns are Agilent PLgel 5 μm MIXED-D,7.5 × 300mm, and the two columns are connected in series; the mobile phase was DMF, the detector was a differential Refractometer (RID), the column temperature was 35 ℃, the flow rate was 0.6ml/min, and the sample volume was 20. mu.l.

The injection results show that the separation degrees of L100-55 and PVP K29/K32, PVP K29/K32 and PEG 400 in the control solution 1 are 3.8 and 1.9 respectively; in the control solution 2, the separation degrees of L100 and PVP K90 and the separation degrees of PVP K90 and PEG600 are respectively 4.2 and 1.7; in the control solution 3, the degrees of separation of L30D-55 from PEG8000 and the degrees of separation of PEG8000 from PVP S630 were 3.7 and 0.6, respectively. The analysis method can realize the simultaneous quantitative analysis of the three components of the acrylic resin, the polyethylene glycol and the povidone.

Example 5: multi-component detection using different mobile phases

(1) Taking 15mg of L100-55, PEG6000 and PVP K30 respectively, placing the L100-55, the PEG6000 and the PVP K30 in the same 20ml measuring flask, preparing 3 parts in parallel, dissolving and diluting the L100-55, the PEG6000 and the PVP K30 to the scale with dimethyl sulfoxide (DMSO), N-methylpyrrolidone (NMP) and dimethylacetamide (DMAc) respectively, and shaking up uniformly to obtain corresponding reference substance solutions.

(2) Measuring the reference substance solution obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result;

wherein, the chromatographic conditions are as follows:

the chromatographic columns are Agilent PLgel 5 μm MIXED-D,7.5 × 300mm, and the two columns are connected in series; the mobile phases are dimethyl sulfoxide (DMSO), N-methyl pyrrolidone (NMP) and dimethyl acetamide (DMAc) respectively, the detectors are Evaporation Light Scattering Detectors (ELSD), the column temperature is 35 ℃, the flow rate is 0.6ml/min, and the sample introduction amount is 20 mu l.

The injection results show that when the mobile phases are dimethyl sulfoxide (DMSO), N-methyl pyrrolidone (NMP) and dimethyl acetamide (DMAc), the simultaneous quantitative analysis of three components of L100-55, PEG6000 and PVP K30 can be realized.

Example 6: multi-component detection using different detectors

(1) Taking 15mg of L100-55, PEG6000 and PVP K30 respectively, placing into a 20ml measuring flask, dissolving with DMF, diluting to scale, and shaking to obtain control solution.

(2) Measuring the reference substance solution obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result;

wherein, the chromatographic conditions are as follows:

the chromatographic columns are Agilent PLgel 5 μm MIXED-D,7.5 × 300mm, two of which are connected in series; the mobile phase was DMF, the detector was an Evaporative Light Scattering Detector (ELSD), the column temperature was 35 deg.C, the flow rate was 0.6ml/min, and the sample volume was 20. mu.l.

The sample injection result shows that when the detector is ELSD, the separation degrees of L100-55 and PEG6000 and the separation degrees of PEG6000 and PVP K30 are respectively 4.0 and 0.7, and the simultaneous quantitative analysis of the three components of L100-55, PEG6000 and PVP K30 can be realized.

Example 7:

(1) appropriate amounts of L100-55, PEG6000 and PVP K30 were placed in a measuring flask to prepare sample solutions of the respective concentrations shown in Table 4.

(2) Measuring the sample solution obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result;

wherein, the chromatographic conditions are as follows:

the chromatographic columns are Agilent PLgel 5 μm MIXED-D,7.5 × 300mm, and the two columns are connected in series; the mobile phase is DMF, the detector is a Refractive Index Detector (RID), the column temperature is 35 ℃, the flow rate is 0.6mL/min, and the sample introduction amount is 20 mu L.

The sample introduction results are shown in Table 4, the separation degrees of L100-55 and PEG6000 are both more than 3.7, the separation degrees of PEG6000 and PVP K30 are both more than 0.5, the recovery rates of the three components are all 93-103%, and the simultaneous quantitative analysis of the three components of L100-55, PEG6000 and PVP K30 can be realized.

TABLE 4 test results of L100-55, PEG6000 and PVP K30 at different concentrations

Example 8:

reference 1[ five peripheral terminals, Zhao Li Na Cao Sha, determination of molecular weight and distribution of acrylic resin [ J ]. Guangzhou chemical industry, 2014,42(24): 103-.

(1) Taking 15mg of L100-55, PEG6000 and PVP K30 respectively, placing into different 20ml measuring flasks, dissolving with Tetrahydrofuran (THF), diluting to scale, and shaking to obtain each component of control solution. Taking 15mg of each of PEG6000, L100-55 and PVP K30, placing in a same 20ml measuring flask, dissolving with Tetrahydrofuran (THF), diluting to scale, and shaking to obtain mixed reference substance solution.

(2) Measuring the reference substance solution and the mixed reference substance solution obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result;

wherein, the chromatographic conditions are as follows:

the chromatographic columns are Agilent PLgel 5 μm MIXED-D,7.5 × 300mm, and the two columns are connected in series; the mobile phase was THF, the detector was a differential Refractive Index Detector (RID), the column temperature and detector temperature were 32 deg.C, the flow rate was 0.3ml/min, and the sample loading was 20. mu.l.

The experiment shows that: PVP K30 is insoluble in THF, so PVP K30 has no peak outflow; ② the PEG6000 has poor solubility in THF, is very slightly soluble in THF (less than 0.4mg/ml), and is not beneficial to the quantitative analysis of PEG 6000.

The chromatogram is shown in figure 7, document 1, PVP K30 is not detected, the peak area of PEG6000 is obviously smaller than that of the method, the separation degree between L100-55 and PEG6000 is poor (the separation degree of the method is 4.6; document 1, the separation degree is 1.2), and the condition can not realize the simultaneous separation and quantitative detection of the three components of L100-55, PEG6000 and PVP K30.

Example 9:

reference 2[ Longjiakun, Jiangjie ice, Zhao Meiyan, etc.. HPLC method determines the content of povidone [ J ] in povidone-iodine oral liquid, J.J.J.Chinese clinical pharmacy, 2016,25(5): 300-302- ].

(1) Taking 0.01g of PVP K30 reference substance, precisely weighing, placing in a 10mL volumetric flask, adding water for dissolving, fixing the volume to a scale, and shaking uniformly to obtain a system applicability solution; taking 15mg of each of PEG6000, L100-55 and PVP K30, respectively placing in different 20ml measuring bottles, dissolving in water, fixing volume to scale, and shaking to obtain each component reference solution; taking 15mg of each of PEG6000, L100-55 and PVP K30, placing in a 20ml measuring flask, adding water to dissolve, fixing volume to scale, and shaking to obtain mixed reference solution.

(2) Measuring the system applicability solution, the component reference substance solution and the mixed reference substance solution obtained in the step (1) by using a high performance liquid chromatography, and analyzing the measurement result;

wherein, the conditions of the high performance liquid chromatography are as follows:

the chromatographic column is Phenomenex C18, 150X 4.6mm, 5 μm; the mobile phase is acetonitrile: water 5: 95; the detector is an ultraviolet detector, and the detection wavelength is 205 nm; the column temperature is 20 ℃; the flow rate is 0.8 ml/min; the sample was taken in an amount of 20. mu.l.

The chromatogram is shown in figure 7, document 2, under the chromatographic conditions, the system applicability passes, no obvious peak appears in L100-55 and PEG6000, and only PVP K30 can be quantitatively detected. Therefore, the condition can not realize the simultaneous quantitative detection of the three components of L100-55, PEG6000 and PVP K30.

Example 10:

reference is made to document 3[ Polyethylene Glycol 3350[ S ]. USP42-NF37,2017:3557 ].

(1) About 0.4g of PEG6000 control was weighed precisely and placed in a 20mL volumetric flask with mobile phase (50. mu.g/mL NaN)₃Aqueous solution), constant volume to scale, shaking up to obtain system applicability solution; taking L100-55, PEG6000 and PVP K30 each 15mg, placing into different 20ml measuring flasks respectively, and adding mobile phase (50 μ g/ml NaN)₃Aqueous solution) and diluted to scale, and shaken up to obtain the control solution of each component. Taking 15mg of each of PEG6000, L100-55 and PVP K30, placing in a 20ml measuring flask, and adding mobile phase (50 μ g/ml NaN)₃Aqueous solution) and diluted to scale, and shaken up to obtain the mixed reference substance solution.

(2) Measuring the system applicability solution, the component reference substance solution and the mixed reference substance solution obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result;

wherein, the chromatographic conditions are as follows:

the chromatographic column is TSK gel G2500PWxl, 7.8X 300mm,7 μm; the mobile phase is 50 mu g/ml NaN₃An aqueous solution; the detector is a differential refraction detector; the column temperature and the detector temperature are 35 ℃; the flow rate is 0.8 ml/min; the sample was taken in an amount of 20. mu.l.

The chromatogram is shown in figure 7, document 3, under the chromatographic conditions, the system applicability is passed, no peak flows out from L100-55, and the chromatographic peaks of PEG6000 and PVP K30 are completely overlapped, so that the chromatographic peaks cannot be separated and quantified.

Example 11:

refer to document 4[ polyethylene glycol residue determination [ S ]. four parts of chinese pharmacopoeia 2015 edition, 2015:238 ].

(1) Taking 5mg of L100-55, PEG6000 and PVP K30 respectively, placing in different 100ml measuring flasks, dissolving with water, diluting to scale, and shaking to obtain control solutions (50 μ g/ml); taking 5mg of each of L100-55, PEG6000 and PVP K30, placing in a 100ml measuring flask, dissolving with water, diluting to scale, shaking to obtain control solution (50 μ g/ml)

(2) Taking 600020 mg of PEG, placing the PEG in a 20ml measuring flask, dissolving the PEG in water, diluting the PEG to a scale, and shaking up to obtain a 1mg/ml sample solution; taking 5ml of 1mg/ml sample solution, placing the sample solution in a 20ml measuring flask, dissolving the sample solution with water, diluting the sample solution to a scale, and shaking up to obtain 250 mu g/ml sample solution; taking 2ml of 1mg/ml sample solution, placing the sample solution in a 20ml measuring flask, dissolving the sample solution with water, diluting the sample solution to a scale mark, and shaking up to obtain the 100 mu g/ml sample solution.

(3) Precisely measuring 1.0ml of the control solution and 1.0ml of the sample solution, adding 5.0ml of 0.5mol/L high chloric acid solution, uniformly mixing, standing at room temperature for 15 minutes, and centrifuging at 4000 rpm for 10 minutes. Taking 4ml of the supernatant, adding 1.0ml of a barium chloride solution (weighing 5g of barium chloride, adding water to dissolve the barium chloride solution to 100ml) and 0.1mol/L of an iodine solution (weighing 2.0g of potassium iodide, adding a small amount of water to dissolve the potassium iodide, then adding 1.3g of iodine, adding water to 50ml, shaking up) to mix evenly, reacting for 15 minutes at room temperature, and measuring absorbance at 535nm by ultraviolet-visible spectrophotometry (0401 in ChP 2015).

The experimental results show that: in the experimental step (2), after the PEG6000 sample solutions of 1mg/ml, 250 μ g/ml and 100 μ g/ml are added into the iodine solution of 0.1mol/L, precipitates are generated, and the larger the concentration is, the more the precipitates are generated, and the absorbance cannot be measured, so that the linear range of the method is judged to be narrower; in addition, PVP K30 and L100-55 also have absorption at the wavelength of 535nm, so that the method is judged to have poor specificity and is easy to be interfered by other components. Namely, the method can not realize the multi-component synchronous separation and quantitative analysis.

Claims (5)

1. A method for simultaneously separating and quantifying acrylic resin, povidone and polyethylene glycol in multiple components is characterized by comprising the following steps:

(1) pretreating a sample to be detected to prepare a sample solution;

(2) measuring the sample obtained in the step (1) by using gel permeation chromatography, and analyzing the measurement result of the sample solution; wherein the mobile phase is N, N-dimethylformamide and does not contain lithium bromide or lithium chloride; the detector is a general detector;

the sample is a multi-component mixture, and the mixture contains any one or combination of more of raw material medicines, triethyl citrate, microcrystalline cellulose, sodium dodecyl sulfate, talcum powder, fumaric acid, crospovidone, saccharin sodium, magnesium stearate, sodium hydroxide, strawberry flavor, liquorice flavor and anhydrous silicon dioxide besides acrylic resin, povidone and polyethylene glycol;

in the step (2), the flow rate of the gel permeation chromatography is 0.2-2 ml/min, the sample injection amount is 5-100 mu l, and the column temperature is 15-40 ℃;

the chromatographic column is Agilent PLgel 5 μm MIXED-D,7.5 × 300mm, and two chromatographic columns or multiple chromatographic columns are connected in series;

common type detectors are differential refractive detectors or evaporative light scattering detectors.

2. The method of claim 1, wherein the acrylic resin type comprises ewing L100-55, ewing L100, ewing L30D-55; the povidone model includes povidone K30, povidone K90, povidone K29/K32, povidone S630; the polyethylene glycol type includes polyethylene glycol 400, polyethylene glycol 600, polyethylene glycol 4000, polyethylene glycol 6000, and polyethylene glycol 8000.

3. The method of claim 1, wherein the step of pre-processing comprises: adding a sample to be detected into a volumetric flask, adding a mobile phase, ultrasonically dissolving or uniformly dispersing the sample, and then carrying out constant volume filtration.

4. The method of claim 3, wherein the sonication time is 5-30 min; after the volume is fixed, the solution is filtered through a polytetrafluoroethylene or equivalent filter membrane with the diameter of 0.22 μm or 0.45 μm.

5. The method according to claim 1, wherein the acrylic resin concentration of the pretreated sample is 0.02-10 mg/ml; the concentration of the povidone is 0.1-10 mg/ml; the concentration of the polyethylene glycol is 0.01-10 mg/ml.

Images