CN113092603A - Method for measuring polysorbate 80 content in Shenmai injection - Google Patents
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Abstract
The invention discloses a method for measuring the content of polysorbate 80 in Shenmai injection, belonging to the technical field of quality detection of traditional Chinese medicine injection. Adding a sodium hydroxide solution into the Shenmai injection, heating to hydrolyze polysorbate 80 to generate oleic acid, removing other components such as traditional Chinese medicine raw materials of the Shenmai injection from the reacted solution by using a solid phase extraction method, only retaining the oleic acid, and measuring the peak area of the oleic acid in a sample by using a high performance liquid chromatography, so that the content of the polysorbate 80 can be obtained according to an external standard method. The method can eliminate interference of other components in the Shenmai injection on determination of polysorbate 80 content, has strong specificity and good stability, and can be used for accurately determining polysorbate 80 content in the Shenmai injection.
Description
Technical Field
The invention belongs to the technical field of quality detection of traditional Chinese medicine injection, and relates to a method for determining the content of polysorbate 80 in Shenmai injection.
Background
There are two types of polysorbate 80 recorded in the "chinese pharmacopoeia", and the polysorbate 80(II) has a higher standard, and has been specified in terms of relative density, viscosity, acid value, saponification value, hydroxyl value, iodine value, peroxide value, ph value, color, ethylene oxide, dioxane, moisture, ignition residue, heavy metal, fatty acid composition, ethylene glycol and diethylene glycol, arsenic salt, freezing test, sterility, bacterial endotoxin, and the like. In the aspect of measuring the content of polysorbate 80, in the Chinese pharmacopoeia, a blue compound is formed according to the reaction of polyethoxy in polysorbate 80 and ammonium cobalt thiocyanate, and the blue compound can be dissolved in dichloromethane and measured by a colorimetric method. However, the components in the traditional Chinese medicine injection are complex, and the interference of negative control is large when the colorimetric method is used for measuring, so that the method is not suitable for measuring the content of polysorbate 80 in the traditional Chinese medicine injection.
In the existing research, the method for measuring the polysorbate 80 content in the traditional Chinese medicine injection mainly comprises a direct measurement method and an indirect measurement method, wherein the direct measurement method comprises a colorimetric method and a gel chromatography method, however, the colorimetric method is easily influenced by other components in the traditional Chinese medicine injection, the polysorbate 80 in the gel chromatography method can generate a plurality of chromatographic peaks, and a large error can be easily generated for the quantification of the polysorbate 80 by using only one chromatographic peak. The indirect determination method comprises a GC method and an HPLC method, wherein polysorbate 80 in the traditional Chinese medicine injection is hydrolyzed into oleic acid, and the oleic acid or methyl oleate is used as a quantitative basis. However, the GC method is complex to operate, and the measurement is mainly carried out by an area normalization method, so that the error is large. Therefore, it is necessary to develop a method for measuring the polysorbate 80 content in the traditional Chinese medicine injection.
Disclosure of Invention
The invention provides a method for measuring the content of polysorbate 80 in Shenmai injection, which hydrolyzes the polysorbate 80 into oleic acid and takes the peak area of the oleic acid as a quantitative basis. Polysorbate 80 is directly used as reference substance, and is treated with SHENMAI injection by the same method, so as to reduce error in measurement process.
The purpose of the invention can be realized by the following technical scheme:
a method for measuring the content of polysorbate 80 in Shenmai injection comprises the following steps:
(1) adding a NaOH solution into the Shenmai injection, heating in a water bath for 30-90 min, cooling, adding an HCl solution and acetonitrile, reacting, and obtaining a mixed solution after the reaction is finished;
(2) transferring the mixed solution in the step (1) to a solid phase extraction column, discarding an effluent liquid, adding a water-acetonitrile mixed solution for leaching, adding acetonitrile for eluting after draining, and collecting an eluent;
(3) and (3) filtering the collected eluent by using a filter membrane, measuring the peak area of an oleic acid absorption peak by using a high performance liquid chromatography, and calculating the content of polysorbate 80 by using an external standard method.
The technical scheme of the invention is as follows: the temperature of the water bath heating in the step (1) is 70-90 ℃.
The technical scheme of the invention is as follows: the volume ratio of the Shenmai injection to the NaOH solution to the HCl solution to the acetonitrile is 1-5: 0.5-1.5: 0.5-1.5: 3-8; wherein: the concentration of the NaOH solution is 0.5-2 mol/L, and the concentration of the HCl solution is 0.5-1.5 mol/L.
In some preferred embodiments: the concentration of the NaOH solution is 0.5-1.5 mol/L.
The technical scheme of the invention is as follows: the solid phase extraction column is a C18 solid phase extraction column with specification of 1 mg.
The technical scheme of the invention is as follows: the C18 solid-phase extraction column is activated by methanol firstly, and then is activated again by 30-80% acetonitrile-water solution.
The technical scheme of the invention is as follows: when the high performance liquid chromatography is used for measurement, the used chromatographic column packing is C18, and the specification of the chromatographic column is 4.6mm multiplied by 250mm and 5 mu m.
The technical scheme of the invention is as follows: when the determination is carried out by using high performance liquid chromatography, the mobile phase is 0.1% phosphoric acid water solution-acetonitrile isocratic elution, the sample amount is 10 mu L, the flow rate is 1.0mL/min, the column temperature is 30 ℃, and the detection wavelength is 203 nm.
In some preferred embodiments: the temperature of water bath heating in the step (1) is 90 ℃, the heating time is 60-65 min, and the concentration of the NaOH solution is 1.05 mol/L.
In some specific embodiments: the invention adopts the alkali complete hydrolysis method and the solid phase extraction technology to determine the content of polysorbate 80 in the Shenmai injection, and the determination method comprises the following steps:
(1) precisely measuring a Shenmai injection, adding 0.5-1.5 mol/L NaOH solution, heating in a water bath at the temperature of 70-90 ℃ for 30-90 min, cooling, adding HCl solution with the same amount as NaOH, adding acetonitrile, and shaking uniformly to obtain a reacted solution;
(2) taking a C18 solid phase extraction column with the specification of 1000mg/6mL, adding 5mL of methanol for activation, and adding 5mL of 50% acetonitrile-water solution for activation;
(3) precisely measuring the reacted solution, transferring the solution to a solid phase extraction column, removing effluent liquid, adding 5-10 mL of water-acetonitrile mixed solution for leaching, and adding acetonitrile with a certain volume for elution after draining;
(4) taking the eluent, filtering with a filter membrane, and measuring the peak area of an oleic acid absorption peak by adopting a high performance liquid chromatography, wherein the filler of a chromatographic column is C18, the specification of the chromatographic column is 4.6mm multiplied by 250mm and is 5 mu m, the mobile phase is 0.1 percent phosphoric acid aqueous solution-acetonitrile (5:95) isocratic elution, the flow rate is 1.0mL/min, the column temperature is 30 ℃, and the detection wavelength is 203 nm. And calculating the content of polysorbate 80 in the Shenmai injection by using polysorbate 80 as a control.
The technical scheme of the invention is as follows: the external standard method comprises the following specific processes:
(1) drawing a standard curve: and precisely weighing and placing the polysorbate-800.2178 g into a 10mL volumetric flask, adding water to the scale, and shaking up to obtain a polysorbate 80 solution. Taking a polysorbate 80 solution, precisely measuring 5mL, placing the solution in a 10mL volumetric flask, adding water to the scale, shaking up to obtain a polysorbate 80 solution diluted by 2 times, sequentially diluting the solution by the same method to obtain polysorbate 80 solutions diluted by 2, 4, 8, 16 and 32 times, operating the same method according to the preparation of a test solution to obtain a series of reference solution, measuring the reference solution according to 2.3 chromatographic conditions, calculating a standard curve of the polysorbate 80 concentration by taking the concentration as x and the peak area as y to obtain a linear equation of y 154897270.18x +21684.14, wherein the linear range is 0.00068-0.02178 g/mL, and R is R2Is 0.9997.
(2) And (3) measuring the sample to be measured by adopting the high performance liquid chromatography to obtain a peak area, and substituting the peak area into the linear equation to obtain the concentration of the sample to be measured.
The invention has the beneficial effects that:
the invention provides a method for measuring the content of polysorbate 80 in Shenmai injection, which comprises the steps of hydrolyzing polysorbate 80 into oleic acid by adopting an alkali complete hydrolysis method as a content measurement index, eliminating the interference of other components in the Shenmai injection by adopting a solid phase extraction technology, and measuring the content of polysorbate 80 in the Shenmai injection.
Drawings
The invention will be further described with reference to the accompanying drawings and examples, in which:
FIG. 1 is an HPLC chromatogram of an oleic acid control solution
FIG. 2 is HPLC chromatogram of hydrolyzed 190711F1 batch Shenmai injection sample
FIG. 3 is a raw HPLC chromatogram of an extract of Shenmai without polysorbate 80
FIG. 4 is a graph of the effect of hydrolysis temperature on polysorbate 80 hydrolysis
FIG. 5 is a graph of the effect of hydrolysis time on polysorbate 80 hydrolysis
FIG. 6 is a graph of the effect of NaOH concentration on polysorbate 80 hydrolysis
FIG. 7 is a three-dimensional graph of the interaction response surface of hydrolysis temperature (A), hydrolysis time (B) and NaOH concentration (C), which is shown in FIG. 7(a), FIG. 7(B) and FIG. 7(C), respectively.
FIG. 8 is HPLC chromatogram of 190711F1 batches of Shenmai injection sample subjected to solid phase extraction
In order to make the technical purpose, technical solutions and technical effects of the present invention more clear and facilitate those skilled in the art to understand and implement the present invention, the present invention will be further described in detail with reference to the accompanying drawings and specific embodiments.
Detailed Description
The invention is further illustrated by the following examples, without limiting the scope of the invention:
example 1 optimization of polysorbate 80 hydrolysis conditions in response to the surface method
Whether polysorbate 80 is completely hydrolyzed or not has an important influence on the experimental results. In order to obtain the optimal polysorbate 80 hydrolysis condition, single-factor investigation is carried out on the main factors influencing hydrolysis, such as hydrolysis temperature, hydrolysis time and sodium hydroxide solution concentration, a Box-Behnken response surface (BBD-RSM) design is adopted, and the hydrolysis condition of polysorbate 80 is optimized by taking the peak area of oleic acid as an index.
1. Material
The instrument comprises the following steps: an OHAUS one-tenth-ten-thousandth electronic balance; HH-4 digital display constant temperature water bath (China electric appliance Co., Ltd.; Waters e2695 high performance liquid chromatograph; Waters 2998PDA detector; Milli-Q Reference ultrapure water system (model: Elix Essential 5UV + Milli-Q Reference).
Reagent: shenmai injection (batch No. 190711F1, 190712F1, 190713F1, 190715F1, 190716F1, Shenwei pharmaceutical industry group Co., Ltd.), Shenmai extract stock solution (batch No. 200725A, provided by Shenwei pharmaceutical industry group Co., Ltd.), polysorbate 80 for injection (batch No. 20190303, Nanjing Wille pharmaceutical industry Co., Ltd.), oleic acid (batch No. L1912119, Aradin reagent (Shanghai) Co., Ltd., HPLC grade), acetonitrile, methanol and phosphoric acid are all chromatographic grades, and other reagents are analytical pure.
2. Method and results
2.1 chromatographic conditions
Dikma Diamonsil C18 chromatographic column (4.6 mm. times.250 mm,5 μm), mobile phase of 0.1% phosphoric acid aqueous solution-acetonitrile (5:95), running time 20min, sample size 10 μ L, flow rate 1.0mL/min, detection wavelength 203nm, column temperature 30 ℃.
2.2 preparation of test solutions
And (3) filtering a reaction solution obtained after the hydrolysis of the polysorbate 80 through a 0.45-micrometer filter membrane to obtain the polysorbate-modified water-based fuel cell.
2.3 preparation of control solutions
Precisely weighing 0.02418g of oleic acid, placing the oleic acid into a 2mL volumetric flask, adding 50% acetonitrile solution to the scale, shaking up, and filtering through a 0.45 mu m filter membrane to obtain an oleic acid reference substance solution. The chromatogram of the oleic acid control is shown in FIG. 1.
2.4 Special Attribute test investigation
Precisely measuring 3.0mL of 190711F1 batches of Shenmai injection, placing the Shenmai injection into a centrifuge tube, precisely adding 1.0mL of 1.0mol/L NaOH solution, sealing, placing the mixture into a water bath at 85 ℃ for heating for 60min, taking out the mixture, cooling the mixture, precisely adding 1.0mL of 1.0mol/L HCl solution, precisely adding 5.0mL of acetonitrile, shaking the mixture evenly, filtering the mixture through a 0.45-micron filter membrane to obtain a sample solution, measuring according to 2.1 chromatographic conditions, and finding a chromatographic peak of the sample at a position corresponding to the chromatogram of a reference substance according to a chromatogram shown in a chromatogram map 2.
The raw liquid of the ginseng and ophiopogon japonicus extractive solution without polysorbate 80 is used as a negative control (figure 3), the method is operated according to the same method, and the detection is carried out according to the 2.1 chromatographic condition, so that no chromatographic peak appears on the negative control at the position corresponding to the chromatogram of the control, and the specificity is good.
2.5 Single factor investigation
2.5.1 Effect of hydrolysis temperature on Polysorbate 80 hydrolysis
Precisely measuring 3.0mL of polysorbate 80 solution, placing in a centrifuge tube, measuring 6 parts, precisely adding 1.0mL of 1.0mol/L NaOH solution, sealing, heating in water bath at 40, 50, 60, 70, 80 and 90 ℃ for 60min, taking out, cooling, precisely adding 1.0mL of 1.0mol/L hydrochloric acid solution, precisely adding 5.0mL of acetonitrile, shaking, filtering with a 0.45-micrometer filter membrane, measuring the area of oleic acid peak, wherein the relationship between the area of oleic acid peak and hydrolysis temperature is shown in FIG. 4.
2.5.2 Effect of hydrolysis time on Polysorbate 80 hydrolysis
Precisely measuring 3.0mL of polysorbate 80 solution, placing in a centrifuge tube, measuring 6 parts, precisely adding 1.0mL of 1.0mol/L NaOH solution, sealing, heating in a water bath at 60 ℃, taking out after 0, 15, 30, 60, 120 and 240min, cooling, precisely adding 1.0mL of 1.0mol/L hydrochloric acid solution, precisely adding 5.0mL of acetonitrile, shaking, filtering with a 0.45-micrometer filter membrane, measuring the oleic acid peak area, wherein the relationship between the oleic acid peak area and hydrolysis time is shown in FIG. 5.
2.5.3 Effect of NaOH concentration on Polysorbate 80 hydrolysis
Precisely measuring 3.0mL of polysorbate 80 solution, placing in a centrifuge tube, measuring 6 parts, precisely adding 1.0mL of NaOH solution of 0.05, 0.1, 0.2, 0.5, 1 and 2mol/L, respectively, sealing, heating in water bath at 60 deg.C for 60min, cooling, precisely adding 1.0mL of hydrochloric acid solution of 0.05, 0.1, 0.2, 0.5, 1 and 2mol/L, precisely adding 5.0mL of acetonitrile, shaking, filtering with 0.45 μm filter membrane, and measuring the area of oleic acid peak, wherein the relationship between the area of oleic acid peak and the concentration of NaOH is shown in FIG. 6.
2.6 response surface design method
According to the results of the single-factor experiment and the response surface analysis method, 3 factors of hydrolysis temperature (A), hydrolysis time (B) and NaOH concentration (C) are respectively selected as independent variables, the oleic acid peak area is used as dependent variable (Y), the Design-expert.8.05b software is used for designing response surface experiments of 3 factor 3 levels, response surface analysis experiments of 17 experimental points are obtained, and the experimental scheme and the results are shown in table 1.
Data were analyzed by Design-expert.8.05b software, and the regression model of the multivariate secondary response surface obtained with oleic acid peak area as the dependent variable Y and 3 independent variables of hydrolysis temperature (a), hydrolysis time (B) and NaOH concentration (C) was Y ═ Y
(16.72+2.301A+1.422B+1.884C-1.273AB-1.631AC-1.122BC-1.000A2-0.828B2-1.083C2)×105，r20.9983, analysis of variance of each factor is shown in table 2. As can be seen, the regression model P<0.0001 significance, statistical significance, mismatching error P0.2064>0.05 was not significant, indicating that the model can be used to fit the experiment. The resulting three-dimensional graph of the response surface is shown in fig. 7(a, b and c). According to the analysis result, the optimal hydrolysis condition is obtained by using Design-expert.8.05b software: the hydrolysis temperature is 90 ℃, the hydrolysis time is 60.48min, and the NaOH concentration is 1.05 mol/L.
TABLE 1 response surface design and results
TABLE 2 response surface quadratic regression equation analysis
Example 2 determination of Polysorbate 80 content in Shenmai injection
1. Material
The instrument comprises the following steps: an experimental instrument: an OHAUS one-tenth-ten-thousandth electronic balance; HH-4 digital display constant temperature water bath (national electric appliance Co., Ltd.); c18 solid phase extraction column (jiangsu green union scientific instruments ltd); waters e2695 hplc; waters 2998PDA detector; the Milli-Q Reference ultrapure water system (model: Elix Essential 5UV + Milli-Q Reference).
Reagent: shenmai injection (batch No. 190711F1, 190712F1, 190713F1, 190715F1, 190716F1, Shenwei pharmaceutical industry group Co., Ltd.), polysorbate 80 for injection (batch No. 20190303, Nanjing Wille pharmaceutical industry Co., Ltd.), Shenmai extract stock solution (batch No. 200725A, provided by Shenwei pharmaceutical industry group Co., Ltd.), acetonitrile and methanol in chromatographic grade, and other reagents in analytical purity.
2. Method of producing a composite material
2.1 preparation of test articles
Precisely measuring 3.0mL of Shenmai injection, placing the Shenmai injection into a centrifuge tube, precisely adding 1.0mL of 1.0mol/L NaOH solution, sealing the tube, placing the tube into a water bath at 90 ℃ for heating for 60min, taking out the tube, cooling the tube, precisely adding 1.0mL of 1.0mol/LHCl solution, precisely adding 5.0mL of acetonitrile, and shaking up to obtain a reaction solution.
Taking a C18 solid phase extraction column with the specification of 1000mg/6mL, adding 5mL of methanol for activation, and adding 5mL of 50% acetonitrile-water solution for balance. Precisely measuring 5.0mL of reaction solution to a solid phase extraction column at a flow rate of 0.5-1 mL/min, adding 5mL of 50% acetonitrile-water solution after the solution is drained, leaching at a flow rate of 1-2 mL/min, and draining when the liquid level flows to the position below a sieve plate. And (3) adding 10mL of acetonitrile for elution, wherein the flow rate is 0.5-1 mL/min, and draining when the liquid level flows below the sieve plate. Collecting eluate, shaking, and filtering with 0.45 μm filter membrane.
2.2 preparation of control solutions
And precisely weighing and placing the polysorbate-800.2178 g into a 10mL volumetric flask, adding water to the scale, and shaking up to obtain a polysorbate 80 solution. Precisely sucking 1.25mL, placing in a 5mL volumetric flask, adding water to the scale, and shaking up. The same method is used for preparing the test solution to obtain the reference solution.
2.3 chromatographic conditions
Dikma Diamonsil C18 chromatographic column (4.6 mm. times.250 mm,5 μm), mobile phase of 0.1% phosphoric acid aqueous solution-acetonitrile (5:95), running time 20min, sample size 10 μ L, flow rate 1.0mL/min, detection wavelength 203nm, column temperature 30 ℃. The chromatogram of the sample of the 190711F1 batch Shenmai injection is shown in figure 8.
2.4 Linear relationship investigation
Taking 5mL of polysorbate 80 solution, precisely measuring the polysorbate 80 solution, putting the solution into a 10mL volumetric flask, adding water to the scale, shaking up to obtain a polysorbate 80 solution diluted by 2 times, sequentially diluting the solution by the same method to obtain polysorbate 80 solutions diluted by 2, 4, 8, 16 and 32 times, operating the solution by the same method according to the preparation of a test solution to obtain a series of reference solution, measuring the solution according to 2.3 chromatographic conditions, calculating a standard curve of the polysorbate 80 concentration by taking the concentration as x and the peak area as y to obtain a linear equation of y-154897270.18 x +21684.14, wherein the linear equation is linearThe range is 0.00068 to 0.02178g/mL, R20.9997, indicating a good linearity.
2.5 precision test investigation
Taking the reference substance solution prepared in the step 2.2, continuously injecting samples for 6 times according to the chromatographic conditions under the item 2.3, determining the peak area of the oleic acid chromatographic peak, and determining that the RSD value of the peak area of the oleic acid chromatographic peak is 0.4 percent, which indicates that the precision is good.
2.6 repeatability test investigation
Precisely measuring 6 parts of the Shenmai injection with the batch number of 190716F1, carrying out the same operation according to the preparation method of a test solution, measuring according to the 2.3 chromatographic condition, calculating the peak area of an oleic acid chromatographic peak, and obtaining a result that the RSD value of the peak area of the oleic acid chromatographic peak is 0.6 percent, thereby showing good repeatability.
2.7 stability test investigation
Taking the prepared product 2.2, and determining the peak area of the oleic acid chromatographic peak in 0, 2, 4, 8, 12 and 24 hours after preparation respectively, wherein the RSD value of the peak area of the oleic acid chromatographic peak is 0.9 percent, which indicates good stability.
2.8 sample application recovery test investigation
Precisely measuring 6 parts of a known-content Shenmai injection, respectively adding an appropriate amount of polysorbate 80 solution (the total amount of the substances to be measured after the addition is equal to 100% of the original amount in the sample), performing the same operation as the preparation method of the sample solution, measuring according to 2.3 chromatographic conditions, and calculating the polysorbate 80 content. The measurement result shows that the sample recovery rate of polysorbate 80 is 103.6%, the RSD is 0.6%, and the sample recovery rate is good.
3. Results
Taking 5 batches of Shenmai injection, preparing according to the method of 2.1, injecting according to the chromatographic condition of 2.3, and determining the peak area of the chromatographic peak of oleic acid. And (3) preparing a polysorbate 80 solution according to a method of 2.1, carrying out sample injection according to a chromatographic condition of 2.3, and determining the peak area of an oleic acid chromatographic peak. The polysorbate 80 content was calculated by external standard method and the results were as follows:
TABLE 3 Tween 80 content in Shenmai injection
The above-mentioned embodiments only express several embodiments of the present invention, and the description is specific and detailed, but not understood as the limitation of the patent scope of the present invention, it should be noted that, for those skilled in the art, it should be noted that, according to the above description, modifications or changes can be made, and all such modifications and changes should fall into the protection scope of the appended claims.
Claims (8)
1. A method for measuring the content of polysorbate 80 in Shenmai injection is characterized in that: the method comprises the following steps:
(1) adding a NaOH solution into the Shenmai injection, heating in a water bath for 30-90 min, cooling, adding an HCl solution and acetonitrile, reacting, and obtaining a mixed solution after the reaction is finished;
(2) transferring the mixed solution in the step (1) to a solid phase extraction column, discarding an effluent liquid, adding a water-acetonitrile mixed solution for leaching, adding acetonitrile for eluting after draining, and collecting an eluent;
(3) and (3) filtering the collected eluent by using a filter membrane, measuring the peak area of an oleic acid absorption peak by using a high performance liquid chromatography, and calculating the content of polysorbate 80 by using an external standard method.
2. The method for measuring the content of polysorbate 80 in the Shenmai injection according to claim 1, wherein the method comprises the following steps: the temperature of the water bath heating in the step (1) is 70-90 ℃.
3. The method for measuring the content of polysorbate 80 in the Shenmai injection according to claim 1, wherein the method comprises the following steps: the volume ratio of the Shenmai injection to the NaOH solution to the HCl solution to the acetonitrile is 1-5: 0.5-1.5: 0.5-1.5: 3-8; wherein: the concentration of the NaOH solution is 0.5-2 mol/L, and the concentration of the HCl solution is 0.5-1.5 mol/L.
4. The method for measuring the content of polysorbate 80 in the Shenmai injection according to claim 1, wherein the method comprises the following steps: the solid phase extraction column is a C18 solid phase extraction column with specification of 1 mg.
5. The method for measuring the content of polysorbate 80 in the Shenmai injection according to claim 4, wherein the method comprises the following steps: the C18 solid-phase extraction column is activated by methanol firstly, and then is activated again by 30-80% acetonitrile-water solution.
6. The method for measuring the content of polysorbate 80 in the Shenmai injection according to claim 1, wherein the method comprises the following steps: when the high performance liquid chromatography is used for measurement, the used chromatographic column packing is C18, and the specification of the chromatographic column is 4.6mm multiplied by 250mm and 5 mu m.
7. The method for measuring the content of polysorbate 80 in the Shenmai injection according to claim 1, wherein the method comprises the following steps: when the determination is carried out by using high performance liquid chromatography, the mobile phase is 0.1% phosphoric acid water solution-acetonitrile isocratic elution, the sample amount is 10 mu L, the flow rate is 1.0mL/min, the column temperature is 30 ℃, and the detection wavelength is 203 nm.
8. The method for measuring the content of polysorbate 80 in the Shenmai injection according to claim 1, wherein the method comprises the following steps: the temperature of water bath heating in the step (1) is 90 ℃, the heating time is 60-65 min, and the concentration of the NaOH solution is 1.05 mol/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115290772A (en) * | 2022-07-18 | 2022-11-04 | 长春圣金诺生物制药有限公司 | Method for detecting polysorbate 80 content in recombinant urate oxidase for injection |
| CN116297915A (en) * | 2023-02-10 | 2023-06-23 | 中国食品药品检定研究院 | Analysis and identification method for polysorbate auxiliary materials |
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