CN113092605A - Analysis method for detecting low molecular weight polyethylene glycol - Google Patents
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Abstract
The invention discloses an analysis method for detecting low molecular weight polyethylene glycol, which is to perform gradient elution by high performance liquid chromatography and a C18 reversed phase chromatographic column by taking a mixed solvent of methanol, tetrahydrofuran and water as a mobile phase and detect by an evaporative light scattering detector. The invention has the advantages of comprehensive information and high separation degree.
Description
Technical Field
The invention relates to an analysis method for detecting low molecular weight polyethylene glycol, and belongs to the field of detection of chemical auxiliary products.
Background
Polyethylene glycol is a chemical auxiliary agent with wide application, and has extremely wide application in the industries of cosmetics, pharmacy, food processing, chemical fiber, plastics, pesticides and the like. In the cosmetics industry, polyethylene glycols of relatively low molecular weight are suitable for use as humectants and consistency regulators, in creams, lotions, toothpaste, shaving creams and the like, and in leave-on hair care preparations to impart silky luster to the hair. In the pharmaceutical industry, low molecular weight polyethylene glycols are widely used in injections, topical preparations, ophthalmic preparations, oral and rectal preparations.
Polyethylene glycol is a polymer of ethylene oxide, and can be classified into a plurality of product categories according to different polymer molecular weights, the molecular weight ranges from 200 to 20000, and common low molecular weight polyethylene glycol mainly comprises several types of PEG200, PEG400, PEG600, PEG800 and PEG1000, and is respectively applied to formula products in different industries.
Polyethylene glycol is not a single-component compound, but a homologous mixture with a certain degree of polymerization distribution, such as PEG600 with an average molecular weight of about 570-630, but the specific components in the product vary widely and greatly according to different manufacturers. The product composition determines the product properties, especially for pharmaceutical formulations, the internal composition of the product directly affects the safety and stability of the final product, so analytical detection of the product is very important for polyethylene glycol. The analytical detection of polyethylene glycol is mainly gel chromatography or chemical analysis, but both methods are apparent analysis methods, i.e. the average molecular weight of a sample can only be measured, and the internal distribution of a product cannot be analyzed. Furthermore, gel chromatography, which is generally accepted to be high, has a poor effect of analyzing substances having a molecular weight of less than 1000, and it is difficult to obtain accurate substance composition information.
Disclosure of Invention
The invention aims to provide an analysis method for detecting low molecular weight polyethylene glycol, which is used for detecting the internal distribution of a polyethylene glycol product with the average molecular weight of 200-1000.
The invention provides an analysis method for detecting low molecular weight polyethylene glycol, which comprises the following steps: analyzing the low molecular weight polyethylene glycol by using liquid chromatography, wherein a C18 reversed phase chromatographic column is adopted as a chromatographic column; the mobile phase comprises A, B two phases, wherein the total amount of the two phases is 100%, the phase A is a mixed solvent of methanol and tetrahydrofuran, the phase B is water, the two phases A, B are mixed according to the volume ratio and then are subjected to gradient elution, and the elution procedure is as follows: the initial proportion of phase A accounts for 5-15%, the rest is phase B, and the phase A gradually transits to phase A accounts for 30-95% and the rest is phase B after 15-35 min; the detector adopts an evaporative light scattering detector; and calculating the relative percentage of each chromatographic peak by an area percentage method, and evaluating the product according to the number and the relative percentage of the chromatographic peaks.
In the above analysis method, the composition of phase a is as follows: a mixed solvent of methanol and tetrahydrofuran, wherein the volume ratio of the tetrahydrofuran is 5-50%, preferably 10-15%;
in the above analytical method, the tetrahydrofuran reagent is preferably a product containing no stabilizer; the atomization temperature of the evaporation light scattering detector is 30-35 ℃, and the evaporation temperature is 80-110 ℃;
in the above analysis method, the low molecular weight polyethylene glycol has the following structural formula:
HO(CH2CH2O)n H
wherein: the average value of n is 4-24, and the average molecular weight of the polyethylene glycol is 200-1000;
the analysis method can be used for carrying out further qualitative and quantitative analysis or relative molecular weight calculation on the target chromatographic peak by using a reference substance or being combined with mass spectrum.
The analysis method can be used for detecting the purity of the product.
The invention has the beneficial effects that:
(1) the invention establishes a new analysis method of the low molecular weight polyethylene glycol, can detect the detailed distribution of the internal components, can specifically determine the relative content of each component, is suitable for the polyethylene glycol product with the average molecular weight of 200-1000, and has the advantages of comprehensive information and high separation degree;
(2) the method can be used for qualitative identification and single-component accurate quantification by matching with a standard reference substance or a mass spectrum detector, and can also be used for average molecular weight calculation;
(3) the method can be used for detecting the purity of the product.
Drawings
FIG. 1: liquid chromatography chromatogram of PEG400 product;
FIG. 2: liquid chromatography chromatogram of PEG600 product;
FIG. 3: liquid chromatography chromatogram of PEG1000 product purity.
Detailed Description
The present invention is further illustrated by, but is not limited to, the following examples.
Example 1:
liquid chromatography analysis of PEG400 product distribution:
the sample was analyzed by liquid chromatography using a C18 column (150 mm. times.4.6 mm,5 μm) and a mobile phase of A, B phases, 100% in total, wherein phase A is a mixed solvent of methanol-tetrahydrofuran (85: 15), the tetrahydrofuran reagent contains no stabilizer, phase B is water, and phase AB is mixed by volume ratio and then eluted by gradient, the elution procedure is: taking 15% of phase A mixed with 85% of phase B as an initial proportion, and transitioning to 40% of phase A mixed with 60% of phase B after 15 min; the flow rate is 1 mL/min-1(ii) a The detector adopts an evaporative light scattering detector, the atomization temperature of the evaporative light scattering detector is 35 ℃, and the evaporation temperature is 90 ℃; the relative percentage content of each chromatographic peak is calculated by an area percentage method, the result is shown in the following table, the product has 12 visible chromatographic peaks, and the content of each peak is basically normally distributed. The chromatogram is shown in figure 1.
The content is as follows:
the sample introduction is carried out for 3 times continuously, the content of the highest chromatographic peak 4 is 23.5%, 23.2% and 23.3% respectively, and the method is proved to have good precision.
Example 2:
liquid chromatography analysis of PEG600 product:
the sample was analyzed by liquid chromatography using a C18 column (100 mm. times.2.1 mm,2.5 μm) and a mobile phase of A, B phases, 100% in total, wherein phase A is a mixed solvent of methanol-tetrahydrofuran (90: 10), the tetrahydrofuran reagent contains no stabilizer, phase B is water, phase AB is mixed by volume ratio and then eluted in a gradient manner, the elution procedure is as follows: taking 10% of phase A mixed with 90% of phase B as an initial proportion, and transitioning to 50% of phase A mixed with 50% of phase B after 20 min; the flow rate was 0.2 mL/min-1Using an evaporative light scattering detectorCarrying out pre-detection, wherein the atomization temperature is 30 ℃, and the evaporation temperature is 100 ℃; 13 peaks of the chromatogram can be seen, and the chromatogram is recorded, which is shown in figure 2. Connecting with a mass spectrum detector, re-injecting sample under the same chromatographic condition for analysis, respectively qualitatively identifying visible chromatographic peaks, assigning each chromatographic peak as follows,
attribution table:
example 3:
liquid chromatography analysis of PEG1000 product purity:
the purity of the sample identified as PEG1000 was analyzed by liquid chromatography using a C18 column (150 mm. times.4.6 mm,5 μm) with a mobile phase of A, B phases, 100% total, wherein phase A is a mixed solvent of methanol-tetrahydrofuran (87: 13), the tetrahydrofuran reagent does not contain a stabilizer, phase B is water, and phase AB is mixed by volume and then eluted by gradient, the elution procedure being: taking 15% of phase A mixed with 85% of phase B as an initial proportion, and transitioning to 90% of phase A mixed with 10% of phase B after 35 min; the flow rate is 1 mL/min-1The detector adopts an evaporative light scattering detector, the atomization temperature is 30 ℃, and the evaporation temperature is 110 ℃; the relative percentage of each chromatographic peak was calculated by area percentage method, see figure 3. The product has two groups of visible chromatographic peaks which are respectively marked as group 1 and group 2, the group 1 has less content, the group 2 has more content, the two groups of chromatographic peaks respectively have independent normal distribution vertexes, and the purity of the sample is determined according to the grouping form and the number of the chromatographic peaks, wherein the relative content of the components in the group 2 is calculated by an area percentage method and is about: 90.7 percent.
Example 4:
liquid chromatography analysis of PEG800 product:
the sample labeled PEG800 was analyzed by liquid chromatography using a C18 column (200 mm. times.4.6 mm,5 μm) and a mobile phase of A, B phases, 100% in total, wherein phase A was a mixed solvent of methanol-tetrahydrofuran (85: 15), the tetrahydrofuran reagent contained no stabilizer, phase B was water, phase A was a mixed solvent of methanol and tetrahydrofuran (85: 15), andmixing the two phases B according to the volume ratio, and performing gradient elution, wherein the elution procedure is as follows: taking 5% of phase A mixed with 95% of phase B as an initial proportion, and transitioning to 25% of phase A mixed with 75% of phase B after 20 min; the flow rate is 1 mL/min-1The detector adopts an evaporative light scattering detector, the atomization temperature is 30 ℃, and the evaporation temperature is 80 ℃; the relative percentage of each chromatographic peak was calculated by area percentage. Taking a PEG800 standard reference substance with known component distribution, carrying out sample injection analysis under the same chromatographic condition, identifying a chromatographic peak of a PEG800 sample to be analyzed by using the chromatographic peak of the reference substance, and calculating the average molecular weight of the sample, wherein the calculation formula is as follows:
M n =∑x i M i
wherein: m n Is the average molecular weight of polyethylene glycol;
x i is the relative percentage content of the component i;
M i is the relative molecular weight of the i component.
The average molecular weight of the PEG800 sample was calculated to be 825.
Example 5:
liquid chromatography analysis of PEG200 product:
the sample identified as PEG200 was analyzed by liquid chromatography using a C18 column (200 mm. times.4.6 mm,5 μm) with a mobile phase of A, B phases, 100% total, wherein phase A is a mixed solvent of methanol-tetrahydrofuran (85: 15), the tetrahydrofuran reagent does not contain a stabilizer, phase B is water, phase AB is mixed by volume and then eluted by gradient, the elution procedure is: taking 5% of phase A mixed with 95% of phase B as an initial proportion, and transitioning to 25% of phase A mixed with 75% of phase B after 30 min; the flow rate is 1 mL/min-1The detector adopts an evaporative light scattering detector, the atomization temperature is 30 ℃, and the evaporation temperature is 80 ℃; the relative percentage of each chromatographic peak was calculated by area percentage. Taking PEG200 standard reference substance with known component distribution, analyzing by sample injection under the same chromatographic condition, identifying the chromatographic peak of PEG200 sample to be analyzed by the chromatographic peak of the reference substance, calculating the average molecular weight of the sample, and calculating the formulaComprises the following steps:
M n =∑x i M i
wherein: m n Is the average molecular weight of polyethylene glycol;
x i is the relative percentage content of the component i;
M i is the relative molecular weight of the i component.
The average molecular weight of the PEG200 sample was calculated to be 205.
Claims (7)
1. An analytical method for the detection of low molecular weight polyethylene glycol, characterized in that: performing gradient elution by high performance liquid chromatography with C18 reversed phase chromatographic column and mixed solvent of methanol, tetrahydrofuran and water as mobile phase, and detecting with evaporative light scattering detector; the low molecular weight polyethylene glycol has the following structural general formula:
HO(CH2CH2O)n H
wherein: the average value of n is 4-24, and the average molecular weight of polyethylene glycol is 200-1100.
2. The analytical method for the detection of low molecular weight polyethylene glycol according to claim 1, characterized by comprising the following steps: analyzing the low molecular weight polyethylene glycol by using liquid chromatography, wherein a C18 reversed phase chromatographic column is adopted as a chromatographic column; the mobile phase comprises A, B two phases, wherein the total amount of the two phases is 100%, the phase A is a mixed solvent of methanol and tetrahydrofuran, the phase B is water, the two phases A, B are mixed according to the volume ratio and then are subjected to gradient elution, and the elution procedure is as follows: the initial proportion of phase A accounts for 5-15%, the rest is phase B, and the phase A gradually transits to phase A accounts for 30-95% and the rest is phase B after 15-35 min; the detector adopts an evaporative light scattering detector; and calculating the relative percentage of each chromatographic peak by an area percentage method, and evaluating the product according to the number and the relative percentage of the chromatographic peaks.
3. The analytical method for the detection of low molecular weight polyethylene glycol according to claim 1, characterized in that: the specific composition of phase A is: the mixed solvent of methanol and tetrahydrofuran, wherein the volume of the tetrahydrofuran accounts for 5-50%.
4. The assay for the detection of low molecular weight polyethylene glycol according to claim 3, characterized in that: the volume ratio of the tetrahydrofuran is 10-15%.
5. The analytical method for the detection of low molecular weight polyethylene glycol according to claim 1, characterized in that: the tetrahydrofuran reagent is a product without a stabilizing agent; the atomization temperature of the evaporation light scattering detector is 30-35 ℃, and the evaporation temperature is 80-110 ℃.
6. The analytical method for the detection of low molecular weight polyethylene glycol according to claim 1, characterized in that: and carrying out further qualitative and quantitative analysis or relative molecular weight calculation on the target chromatographic peak by using a reference substance or being combined with the mass spectrum.
7. Use of the analytical method according to any one of claims 1 to 6 for the detection of low molecular weight polyethylene glycol according to claim 1 for the detection of product purity.
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| CN114062560A (en) * | 2021-11-29 | 2022-02-18 | 中钢集团鞍山热能研究院有限公司 | High performance liquid chromatography analysis method for distribution of components with different polymerization degrees of phthalonitrile resin |
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