Method for determining contents of different polysorbates

Abstract

The invention discloses a method for determining different polysorbate contents, which comprises the following steps: S1, diluting polysorbate series standard substances to prepare standard solutions, injecting the standard solutions into a liquid chromatograph for detection, and drawing a standard curve by taking the logarithm value of the concentration of the standard solutions as the x axis and the logarithm value of the peak area of liquid chromatography corresponding to the standard solutions as the y axis; s2, diluting a polysorbate sample to obtain a test solution 1, and diluting a preparation sample containing polysorbate to obtain a test solution 2; injecting the test solution 1 and the test solution 2 into a liquid chromatograph for detection to obtain corresponding chromatograms; and S3, calculating the content of the characteristic substance in the test solution 1 according to the standard curve and the peak area of the characteristic substance in the chromatogram obtained from the test solution 1, and calculating the content of polysorbate in the test solution 2 according to the peak area of the characteristic substance in the chromatogram obtained from the test solution 2. According to the invention, the variety and content of polysorbate series can be simply, conveniently and accurately detected.

Description

Technical Field

[0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to a method for determining the contents of different polysorbates. Background Art

[0002] Polysorbate is a high molecular polymer formed by the polymerization of fatty acids, sorbitol, dehydrated products and ethylene oxide. It can be divided into polysorbate 20, 40, 60 and 80 series according to the different fatty acid compositions. The fatty acids in its structure are lauric acid (C12), palmitic acid (C16), stearic acid (C18), oleic acid (C18 with one unsaturated olefin bond), linoleic acid (C18 with two unsaturated olefin bonds), etc. It is used as wetting agents, solubilizers, emulsifiers and protein stabilizers in preparations.

[0003] There are two main production processes for polysorbates: one is ester first and then ether (fatty acids are first polymerized with sorbitol and its dehydrated products and ethylene oxide to form esters, and then polymerized with fatty acids); the other is ether first and then ester (sorbitol and its dehydrated products are first polymerized with ethylene oxide, and then esterified with fatty acids, and subsequent refining treatment processes.

[0004] The methods reported for determining the content of polysorbate are: 1. UV method. The principle of this method is to use polysorbate and cobalt thiocyanate for color development and then perform the determination. The disadvantage of this method is that there are many interference factors; 2. Polysorbate standard is hydrolyzed and methylated. The test sample is also hydrolyzed and methylated. The content of polysorbate is calculated by the content of fatty acid methyl ester. However, this method requires certain pre-treatment steps and is cumbersome to operate. When the preparation contains other ingredients with fatty acid structure, the content of polysorbate cannot be determined; 3. . The content of polysorbate can also be determined by quadrupole mass spectrometry, collecting chromatographic peaks of corresponding molecular weights, thereby calculating the content of polysorbate in the test sample. This method requires a quadrupole mass spectrometer as a detector; 4. The content of polysorbate can also be determined by high performance liquid chromatography using a fluorescence detector, but the disadvantage of this method is that antibodies will interfere and it is impossible to distinguish between polysorbate 20 or 80 series products; 5. Use a spectrometer to test the infrared attenuated total reflection spectrum, and identify the wave number corresponding to the C=O peak from the reflection spectrum (for example, 1735cm -1 ), identifying the wavenumber corresponding to the internal standard from the reflectance spectrum, calculating the area under the curve of the C=O wavenumber normalized by the internal standard, and comparing the area with the standard curve to determine the concentration of polysorbate in the sample; this method is targeted at the C=O structure in the polysorbate structure, so other non-polysorbate series with C=O structure will also be included, affecting its accuracy.

[0005] None of the above methods can accurately distinguish the types of polysorbates. Some methods have complex pretreatments or require a quadrupole mass spectrometer as a detector.

[0006] Since all polysorbate series contain a certain amount of free polyethylene glycol, the content will vary due to different synthesis processes, which does not provide much guidance for distinguishing different polysorbate products. However, the types and contents of fatty acids in the starting materials of polysorbate series products vary greatly, resulting in the existence of exclusive components in the polysorbate finished products. This component has great guidance for distinguishing the polysorbate series, and the type and prescription dosage of polysorbate in the preparation can be calculated based on the content of its specific components in the polysorbate series. Summary of the invention

[0007] The primary purpose of the present invention is to provide a method for determining the content of different polysorbates, so as to be able to simply and accurately detect the polysorbate series and content, thereby ensuring consistency with the original formulation and ensuring the safety of the preparation.

[0008] To this end, the present invention provides the following technical solutions.

[0009] One aspect of the present invention provides a method for determining the content of different polysorbates, the method comprising the following steps: S1: diluting a polysorbate series standard substance to prepare a standard solution, injecting the solution into a liquid chromatograph for detection, and plotting a standard curve with the logarithm of the concentration of the standard solution as the x-axis and the logarithm of the liquid chromatographic peak area corresponding to the standard solution as the y-axis; S2: diluting the polysorbate sample to obtain a test solution 1, and diluting the preparation sample containing polysorbate to obtain a test solution 2; injecting the test solution 1 and the test solution 2 into a liquid chromatograph for detection to obtain a corresponding chromatogram; S3: Calculate the content of the characteristic substance in test solution 1 based on the standard curve and the peak area of ​​the characteristic substance in the chromatogram obtained from test solution 1, and calculate the content of polysorbate in test solution 2 based on the peak area of ​​the characteristic substance in the chromatogram obtained from test solution 2.

[0010] In a preferred embodiment of the present invention, in step S1, the polysorbate series standard substances include specific components of polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80, such as sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate and sorbitan monooleate.

[0011] In a preferred embodiment of the present invention, in steps S1 and S2, the stationary phase of the liquid chromatograph uses a reverse phase chromatography system, and the detector used is an evaporative light scattering detector.

[0012] In a preferred embodiment of the present invention, the reverse phase chromatography column of the reverse phase chromatography system uses reverse phase carbon octadecylsilane bonded silica gel as a filling column, and uses acetonitrile containing a certain acid as mobile phase B and water containing a certain acid as mobile phase A for gradient elution.

[0013] In a preferred embodiment of the present invention, the packed column has a length of 50 mm, an inner diameter of 4.6 mm, and a particle size of 3.5 µm.

[0014] In a preferred embodiment of the present invention, the acid is formic acid or acetic acid, and the usage amount of the acid is 0.02-0.08%.

[0015] In a preferred embodiment of the present invention, the gradient elution program is: in terms of time min / mobile phase B%, it is 0 / 15, 16 / 100, 26 / 100, 17 / 15, 30 / 15 respectively, the flow rate is 1.0~1.5ml / min, and the column temperature is 45~55℃.

[0016] In a preferred embodiment of the present invention, the drift tube temperature of the evaporative light scattering detector is 75-85° C., the carrier gas flow rate is 1.2-1.8 L / min, and the gain is 1.0-2.5.

[0017] In a preferred embodiment of the present invention, in step S2, the sample to be tested is an injection, oral solution, emulsion or other oral solid dosage form containing one or more of polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80.

[0018] In a preferred embodiment of the present invention, in step S3, a logarithmic regression equation is used to quantify the sample to be tested.

[0019] By means of the above technical scheme, the present invention has at least the following advantages: the quantitative analysis method of the present invention has high sensitivity, good repeatability and precision, and has the advantages of being fast, efficient and simple, and is suitable for determining the content of polysorbate series excipients in different preparations, and can effectively supervise production and ensure the stability of product quality.

[0020] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the preferred embodiments of the present invention are described in detail as follows. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is the liquid chromatogram of 2mg / ml polyoxyethylene 20 sorbitan monolaurate standard; Figure 2 This is the liquid chromatogram of the 2 mg / ml polysorbate 20 test sample; Figure 3is the liquid chromatogram of 0.5 mg / ml polyethylene glycol 1000 standard; Figure 4 This is the liquid chromatogram of the 2 mg / ml polysorbate 80 test sample; Figure 5 This is the liquid chromatogram of the XTLQ eye drops test sample. DETAILED DESCRIPTION

[0022] In order to make the technical means, creative features, objectives and effects of the present invention easy to understand, the technical solutions in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] Unless otherwise specified, the percentage content involved in the present invention refers to mass percentage for solid-liquid mixing and solid-solid mixing, and refers to volume percentage for liquid-liquid mixing.

[0024] Unless otherwise specified, the percentage concentrations referred to in the present invention all refer to the final concentration, which refers to the percentage of the added component in the system after the addition of the component.

[0025] The temperature parameters in the present invention, if not specifically limited, allow for both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows the temperature to fluctuate within the precision range controlled by the instrument.

[0026] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0027] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0028] Unless otherwise specified, the following liquid chromatography method is carried out in accordance with the liquid chromatography method in Part IV of the Chinese Pharmacopoeia 2020 edition.

[0029] The present invention provides a method for rapidly detecting the content of polysorbate in a preparation by using a reversed-phase high performance liquid chromatography method, which is applicable to the quantitative detection of a series of products. The method can be applied to the analysis and detection of the content of polysorbate in injections, fat emulsions, oral solutions and other oral solid preparations in the fields of medicines and cosmetics.

[0030] In the liquid chromatography method used in the present invention, the stationary phase uses a reverse phase chromatography system, the detector is an evaporative light scattering detector, and the logarithmic regression equation is used for quantification, wherein the chromatographic conditions are: a. Octadecylsilane bonded silica gel is used as the filling column; the column length is 50 mm, the inner diameter is 4.6 mm, and the particle size is 3.5 µm.

[0031] b. Gradient elution was performed with an acidic aqueous solution as mobile phase A and acidic acetonitrile as mobile phase B; the acid used in this embodiment was acetic acid, and the amount added was 0.05%.

[0032] c. Gradient elution program (time min / b%): 0 / 15, 16 / 100, 26 / 100, 27 / 15, 37 / 15; d. Flow rate: 1.5 ml / min; e. Column temperature 50℃; f. The drift tube temperature of the evaporative light scattering detector is 80°C, the carrier gas flow rate is 1.5 L / min, and the gain is 1.5.

[0033] In the embodiment of the present invention, a standard curve solution is first prepared, and the specific operation is as follows: Standard curve 1: Take an appropriate amount of characteristic standard substance in polysorbate, weigh accurately, and use acetonitrile-water to prepare solutions with concentrations of 0.01 mg / ml, 0.05 mg / ml, 0.1 mg / ml, 0.3 mg / ml, 0.5 mg / ml, and 0.7 mg / ml as standard curve 1 solutions.

[0034] Preparation of test solution 1: Take 0.2 g of polysorbate sample, accurately weigh it, place it in a 100 ml volumetric flask, dissolve it in acetonitrile-water (1:1) and dilute it to the scale, shake well, and use it as test solution 1.

[0035] Standard curve 2: Take an appropriate amount of polysorbate sample and use acetonitrile-acetonitrile-water (1:1) to prepare solutions with concentrations of 0.5 mg / ml, 0.75 mg / ml, 1.0 mg / ml, 1.5 mg / ml, and 2.0 mg / ml, respectively, as standard curve 2 solutions.

[0036] Test solution 2: XTLQ eye drops from batch 2022080201 were directly used as test solution 2.

[0037] Determination method: Accurately measure the standard curve 1 solution and the test sample solution 1 respectively, inject them into the liquid chromatograph for determination, and the content of the characteristic component in the test sample 1 can be calculated from the standard curve 1; take the standard curve 2 and the test sample 2, inject them into the liquid chromatograph for determination, and the amount of polysorbate in the test sample 2 can be calculated from the standard curve 2.

[0038] Concentration calculation method: Use the logarithm of the peak area of ​​the standard curve solution as the y-axis and the logarithm of the concentration of the standard curve solution as the x-axis to perform linear regression and obtain a linear equation.

[0039] Example 1: Determination of the content of sorbitan monolaurate, a specific component in polysorbate 20 A. Sample preparation Preparation of standard curve 1 solution: Take 0.4g of polyoxyethylene 20 sorbitan monolaurate reference substance, weigh accurately, place in a 20ml volumetric flask, dissolve with acetonitrile-water (1:1), dilute to scale, and shake well as standard stock solution. Take appropriate amount of standard stock solution and dilute with acetonitrile-water (1:1) to concentrations of 0.01mg / ml, 0.05mg / ml, 0.1mg / ml, 0.3mg / ml, 0.5mg / ml, 0.7mg / ml as standard curve solution.

[0040] Preparation of test solution 1: Taking polysorbate 20 as an example, take 0.2 g of polysorbate 20 sample, accurately weigh it, place it in a 100 ml volumetric flask, dissolve it with acetonitrile-water (1:1) and dilute it to the scale, and shake it well as the test solution 1.

[0041] The chromatographic conditions were as follows: an octadecylsilane bonded silica gel column was used as the chromatographic column (50 mm × 4.6 mm, 3.5 µm), 0.05% formic acid water was used as the mobile phase A, 0.05% formic acid acetonitrile was used as the mobile phase B for gradient elution, the flow rate was 1.5 ml / min, the column temperature was 50 °C, the drift tube temperature of the evaporative light scattering detector was 80 °C, the carrier gas flow rate was 1.5 L / min, and the gain was 2.0. 10 μl of the standard curve solution and the test solution 1 and 2 were injected into the liquid chromatograph for determination.

[0042] Gradient elution program (time min / b%): 0 / 15, 16 / 100, 26 / 100, 27 / 15, 37 / 15.

[0043] B. Determination of linear range In this embodiment, the linear range of the method of the present invention for detecting different concentrations of polysorbate was explored. The specific operation method is: take polyoxyethylene 20 sorbitan monolaurate standard and dilute it to concentrations of 0.01mg / ml, 0.05mg / ml, 0.1mg / ml, 0.3mg / ml, 0.5mg / ml, and 0.7mg / ml as standard curve solutions. Take 20µl of the standard curve solutions of each concentration and inject them into a liquid chromatograph for detection to obtain the corresponding chromatogram. Take the logarithm of the concentration C (mg / ml) of the standard curve solution as the horizontal coordinate X, and take the logarithm of the peak area A in the chromatogram as the vertical coordinate Y for linear regression. Obtain a standard curve.

[0044] The obtained standard curve equation is: y=1.7501x+0.5158, and the correlation coefficient r is 0.9994.

[0045] Acceptance criteria: The logarithm of the concentration is the horizontal coordinate x, and the logarithm of the peak area is the vertical coordinate y: the correlation coefficient (r) must not be less than 0.995, and the Y-axis intercept should be within 25% of the 100% response value.

[0046] The above results show that when the concentration of polyoxyethylene (20) sorbitan monolaurate is between 0.01 and 0.7 mg / ml, the linear relationship between the logarithm of the peak area and the logarithm of the concentration is good.

[0047] C. Limit of detection and limit of quantification The linear concentration of polyoxyethylene (20) sorbitan monolaurate was 0.01 mg / ml.

[0048] Accurately take 3 ml of polyoxyethylene (20) sorbitan monolaurate linear 1 solution and dilute it to 10 ml, which is the detection limit concentration, that is, the detection limit concentration is 0.003 mg / ml.

[0049] D. Precision experiment Take a solution with a concentration of 0.3 mg / ml in the standard curve solution of polyoxyethylene 20 sorbitan monolaurate, inject it into a liquid chromatograph for detection, and obtain the corresponding chromatogram. Among them, the injection precision is 6 injections in a row; In addition, polysorbate 20 test solution 1 (6 portions, 1 injection each) was taken as repeatability; The intermediate precision was obtained by performing repeated experiments again by different experimenters at different times.

[0050] The RSD of the logarithm of the peak area of ​​6 injections of 0.3 mg / ml of the present invention is 1.5%; the average content of polyoxyethylene (20) sorbitan monolaurate in the polysorbate 20 of the present invention is 10.18%, the RSD is 1.2%, the average of the intermediate precision of 12 parts is 10.35%, and the RSD is 2.6%; the average content of polysorbate 20 in the eye drops is 0.11%, the RSD is 3.8%, the average of the intermediate precision of 12 parts is 0.12%, and the RSD is 6.5%. The above results show that the method of the present invention has good precision.

[0051] E: Recovery rate test Take an appropriate amount of polysorbate 20, add acetonitrile-water (1:1) to dissolve it, then add a certain amount of polyoxyethylene (20) sorbitan monolaurate reference solution according to the actual measured amount of 50%, 100% and 150%, inject it into the liquid chromatograph for detection, operate 3 times in parallel on each level, and collect chromatograms.

[0052] Substitute the corresponding peak area in the chromatogram of the test sample into the above standard curve equation, calculate the concentration as the measured concentration, take the ratio of the measured concentration to the accurate concentration added as the recovery rate, and calculate the recovery amount and recovery rate.

[0053] The results showed that the recovery rates of the present invention were between 95% and 105%, and the RSD was 4.8%, indicating that the present invention had good accuracy.

[0054] F: Durability determination 1. Solution stability Take 100% of the test solution added under the accuracy item (the background concentration in the test sample is 0.2 mg / ml, add an equal amount of reference substance, the total concentration is 0.4 mg / ml), place it at room temperature, and inject it into the liquid chromatograph for determination at 0, 2, 4, 6, 8, 12 and 24 h respectively.

[0055] The results showed that the RSD of the logarithm of the peak area of ​​the 100% spiked test solution placed at room temperature for at least 24 hours was 1.8%, indicating that the test solution was stable when placed at room temperature for at least 24 hours.

[0056] 2. Change the chromatographic column Replace the chromatographic columns with different numbers (9912112028 and 9912112171) but with the same filler and the same specifications for determination.

[0057] The results show that when the chromatographic column of the same specification from the same manufacturer is replaced, the average content of polyoxyethylene (20) sorbitan monolaurate in polysorbate 20 is measured to be 10.21%, indicating that the batch number of the chromatographic column has little effect on the present invention.

[0058] 3. Change flow rate The flow rate was changed to 1.4 and 1.6 ml / min for liquid chromatography determination.

[0059] The results showed that when the flow rate was changed to 1.4 ml / min, the content of polyoxyethylene (20) sorbitan monolaurate in polysorbate 20 was measured to be 10.32%; when the flow rate was changed to 1.6 ml / min, the content of polyoxyethylene (20) sorbitan monolaurate in polysorbate 20 was measured to be 10.19%, indicating that the carrier gas flow rate has little effect on the present invention.

[0060] 4. Change column temperature The column temperature was changed to 45°C and 55°C for liquid chromatography determination.

[0061] The results show that when the column temperature is 45°C, the content of polyoxyethylene (20) sorbitan monolaurate in polysorbate 20 is measured to be 10.31%; when the column temperature is 55°C, the content of polyoxyethylene (20) sorbitan monolaurate in polysorbate 20 is measured to be 10.13%, indicating that the column temperature has little effect on the results of the present invention.

[0062] 5. Slightly adjust the initial proportion of acetonitrile in the mobile phase for liquid chromatography determination.

[0063] The results showed that when the initial ratio of acetonitrile was 13, the content of polyoxyethylene (20) sorbitan monolaurate in polysorbate 20 was measured to be 10.45%; when the initial ratio was 17, the content of polyoxyethylene (20) sorbitan monolaurate in polysorbate 20 was measured to be 10.24%, indicating that the initial ratio of acetonitrile in the mobile phase had almost no effect on the results.

[0064] The above results show that the present invention has good durability.

[0065] G: Sample determination Different batches of polysorbate 20 were taken as test samples, and the content of polyoxyethylene (20) sorbitan monolaurate therein was determined according to the method of the present invention. The results showed that the content was between 9% and 13%, indicating that the content of this component was relatively stable in different synthesis processes.

[0066] Implementation Case 2: Detection of Polysorbate 80 in Eye Drops Preparation of standard curve 2 solution: Take an appropriate amount of polysorbate 80, weigh accurately, dissolve and dilute with water to a solution of 10 mg / ml, and shake well as the standard stock solution. Take an appropriate amount of the standard stock solution and dilute with acetonitrile-water (1:1) to concentrations of 0.25 mg / ml, 0.5 mg / ml, 1.0 mg / ml, 1.5 mg / ml, and 2.0 mg / ml as the standard curve 2 solution.

[0067] Preparation of test solution 2: Taking eye drops as an example, take 1 g of XTLQ eye drops from batch 2022080201 (adjust according to the prescription amount), place it in a 10 ml volumetric flask, dissolve and dilute to the scale with acetonitrile-water (1:1), and shake well as test solution 2.

[0068] A standard curve was drawn with the logarithm of the peak area of ​​the specific component with a retention time of about 19 min in the chromatogram as the ordinate and the logarithm of the concentration of polysorbate 80 as the abscissa. The equation of the standard curve was y=1.5858x+1.1477. The peak area of ​​the component in the titration solution was substituted into the standard curve, and the amount of polysorbate 80 in the 2022080201 batch of XTLQ eye drops was obtained to be 1.0 mg / ml.

[0069] The invention can conveniently, quickly and efficiently detect the types and contents of polysorbates in different preparations, wherein different polysorbate products are prepared with standard curves using different characteristic impurity reference substances for determination.

[0070] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technician familiar with this profession can make some changes or modifications to equivalent embodiments of equivalent changes by using the methods and technical contents disclosed above without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A method for determining the content of different polysorbates, characterized in that: The method comprises the following steps: S1: diluting a polysorbate series standard substance to prepare a standard solution, injecting the solution into a liquid chromatograph for detection, and plotting a standard curve with the logarithm of the concentration of the standard solution as the x-axis and the logarithm of the liquid chromatographic peak area corresponding to the standard solution as the y-axis; S2: diluting the polysorbate sample to obtain a test solution 1, and diluting the preparation sample containing polysorbate to obtain a test solution 2; injecting the test solution 1 and the test solution 2 into a liquid chromatograph for detection to obtain a corresponding chromatogram; S3: Calculate the content of the characteristic substance in test solution 1 based on the standard curve and the peak area of ​​the characteristic substance in the chromatogram obtained from test solution 1, and calculate the content of polysorbate in test solution 2 based on the peak area of ​​the characteristic substance in the chromatogram obtained from test solution 2.

2. The method for measuring different polysorbate contents according to claim 1, characterized in that: In step S1, the polysorbate series standard substances include polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80.

3. The method for measuring different polysorbate contents according to claim 1, characterized in that: In steps S1 and S2, the stationary phase of the liquid chromatograph uses a reverse phase chromatography system, and the detector used is an evaporative light scattering detector.

4. The method for measuring different polysorbate contents according to claim 3, characterized in that: The reverse phase chromatography column of the reverse phase chromatography system uses reverse phase carbon octadecylsilane bonded silica gel as a filling column, and uses acetonitrile containing a certain acid as a mobile phase B and water containing a certain acid as a mobile phase A for gradient elution.

5. The method for measuring different polysorbate contents according to claim 4, characterized in that: The column length of the packed column is 50 mm, the inner diameter is 4.6 mm, and the particle size is 3.5 μm.

6. The method for measuring different polysorbate contents according to claim 4, characterized in that: The acid is formic acid or acetic acid, and the usage amount of the acid is 0.02-0.08%.

7. The method for measuring different polysorbate contents according to claim 4, characterized in that: The gradient elution program is: in terms of time min / B%, it is 0 / 15, 16 / 100, 26 / 100, 17 / 15, 30 / 15 respectively, the flow rate is 1.0-1.5 ml / min, and the column temperature is 45-55°C.

8. The method for measuring different polysorbate contents according to claim 3, characterized in that: The drift tube temperature of the evaporative light scattering detector is 75-85° C., the carrier gas flow rate is 1.2-1.8 L / min, and the gain is 1.0-2.

5.

9. The method for measuring different polysorbate contents according to claim 1, characterized in that: In step S2, the sample to be tested is an injection, oral solution, emulsion or other oral solid dosage form containing one or more of polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80.

10. The method for determining different polysorbate contents according to claim 1, characterized in that: In step S3, the logarithmic regression equation is used to quantify the sample to be tested.

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