CN117871752A - Quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in biological medicine stock solution - Google Patents
Quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in biological medicine stock solution Download PDFInfo
- Publication number
- CN117871752A CN117871752A CN202410063462.1A CN202410063462A CN117871752A CN 117871752 A CN117871752 A CN 117871752A CN 202410063462 A CN202410063462 A CN 202410063462A CN 117871752 A CN117871752 A CN 117871752A
- Authority
- CN
- China
- Prior art keywords
- ethylene oxide
- solution
- polyoxypropylene ethylene
- mobile phase
- glycerol ether
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- -1 polyoxypropylene ethylene oxide glycerol Polymers 0.000 title claims abstract description 62
- RBNPOMFGQQGHHO-UHFFFAOYSA-N glyceric acid Chemical group OCC(O)C(O)=O RBNPOMFGQQGHHO-UHFFFAOYSA-N 0.000 title claims abstract description 35
- 238000001514 detection method Methods 0.000 title claims abstract description 21
- 239000003814 drug Substances 0.000 title claims abstract description 20
- 239000011550 stock solution Substances 0.000 title claims abstract description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 42
- 239000004721 Polyphenylene oxide Substances 0.000 claims abstract description 39
- 229920000570 polyether Polymers 0.000 claims abstract description 39
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000000523 sample Substances 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 20
- 238000012360 testing method Methods 0.000 claims abstract description 20
- 239000012488 sample solution Substances 0.000 claims abstract description 19
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 14
- 239000012498 ultrapure water Substances 0.000 claims abstract description 14
- 238000010828 elution Methods 0.000 claims abstract description 10
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 34
- 238000001914 filtration Methods 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 238000007865 diluting Methods 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 239000012086 standard solution Substances 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 239000003085 diluting agent Substances 0.000 claims description 4
- 238000010812 external standard method Methods 0.000 claims description 4
- 239000012088 reference solution Substances 0.000 claims description 3
- 239000012085 test solution Substances 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 2
- 238000005070 sampling Methods 0.000 claims description 2
- 229940079593 drug Drugs 0.000 claims 5
- GPLRAVKSCUXZTP-UHFFFAOYSA-N diglycerol Chemical compound OCC(O)COCC(O)CO GPLRAVKSCUXZTP-UHFFFAOYSA-N 0.000 abstract description 27
- 238000011084 recovery Methods 0.000 abstract description 9
- 238000000926 separation method Methods 0.000 abstract description 5
- 238000005259 measurement Methods 0.000 abstract description 4
- 230000008569 process Effects 0.000 abstract description 3
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 230000007246 mechanism Effects 0.000 abstract description 2
- 239000013558 reference substance Substances 0.000 description 10
- 238000011002 quantification Methods 0.000 description 5
- 230000014759 maintenance of location Effects 0.000 description 4
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 229960000074 biopharmaceutical Drugs 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 239000013530 defoamer Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000003505 mutagenic effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 238000010953 Ames test Methods 0.000 description 1
- 231100000039 Ames test Toxicity 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010028400 Mutagenic effect Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 150000004945 aromatic hydrocarbons Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000012490 blank solution Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 210000002798 bone marrow cell Anatomy 0.000 description 1
- 238000009924 canning Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 231100000243 mutagenic effect Toxicity 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- 150000002894 organic compounds Chemical group 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in biological medicine stock solution. The biological medicine stock solution dissolved in acetonitrile is selected as a sample solution, and is detected by adopting a high performance liquid chromatography, and a C-18 reversed phase chromatographic column with the diameter of 4.6X250 mm is used; mobile phase A is methanol, and mobile phase B is laboratory ultrapure water for gradient elution. Mobile phase a: methanol, mobile phase B: the laboratory ultrapure water determination method is based on a separation mechanism of high performance liquid chromatography, adopts a novel universal detector, namely an electrospray detector CAD, has high detection sensitivity and good repeatability, ensures the stability of a sample in the test process, and realizes the scientific determination of polyoxypropylene ethylene oxide glyceryl ether polyether residues. The method is evaluated by the recovery rate and the relative standard deviation, the recovery rate is measured to be 95-106%, and the accuracy is good; the relative standard deviation of the parallel measurement of the samples is less than 2.0%, and the precision of the method is good.
Description
Technical Field
The invention belongs to the technical field of polyoxypropylene ethylene oxide glycerol ether detection, and particularly relates to a quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in biological medicine stock solution.
Background
Polyoxypropylene ethylene oxide glyceryl ether is an organic compound, alias: GPE defoamers are colorless or yellow non-volatile oily liquids at normal temperature. The relative density is 1.095g/mLat25 ℃, the melting point is 60-50 ℃, the boiling point is >200 ℃ (lit.) and the flash point is >230 DEG F, the refractive index n20/D is 1.466, the pH is 5.0-7.5 (100 g/LinH 2O), the storage condition is 2-8 ℃, and the solvent is dissolved in benzene and other aromatic hydrocarbon solvents, and also dissolved in solvents such as diethyl ether, ethanol, acetone, carbon tetrachloride and the like. The chemical formula is:
biopharmaceuticals refer to a broad class of products for prevention, diagnosis and treatment manufactured using organisms, biological tissues or components thereof, and by combining the principles and methods of biology, biochemistry, physicochemical, biotechnology and pharmacy. The broad sense of biopharmaceuticals includes various natural bioactive substances and artificially synthesized or semisynthetic analogues of natural substances prepared from organisms such as animals, plants, microorganisms and the like. In the production process of biological medicine, many processes can generate bubbles, so that the problems of liquid leakage and the like are caused, and the canning amount of the reaction tank is reduced, or the stirring is reduced or stopped, so that the fermentation efficiency and the productivity are affected. The polyoxypropylene ethylene oxide glycerol ether is a common small molecule defoamer, but due to the fact that the defoamer has certain toxicity, related researches show that: LD50 mice were orally administered 379.40mg/kg (bw), accumulated toxicity K value was greater than 5.3, mutagenic test Ames test, mouse bone marrow cell micronucleus test and mouse sperm aberration test, all without mutagenic effect.
And the polyoxypropylene ethylene oxide glycerol ether is removed in the subsequent production process of the biological medicine stock solution, and the residual quantity in the stock solution is quantitatively detected, so that the quality of a final product is ensured.
The current standards of polyoxypropylene ethylene oxide glycerol ether only include national food safety standards of food additive polyoxypropylene ethylene oxide glycerol ether, and the standards also have no detection of the content of polyoxypropylene ethylene oxide glycerol ether, and the specific reference is GB1886.298-2018. However, the method for detecting the polyoxypropylene ethylene oxide glyceryl ether as a residue is not yet available.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention aims to provide a quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues. The detection method has the advantages of stable detection result, good reproducibility of samples detected by different operators at different times and simple operation.
In order to achieve the above purpose, the present invention provides the following technical solutions: a quantitative detection method of polyoxypropylene ethylene oxide glycerol ether residues comprises the following steps:
step one, preparing a reagent for testing, wherein the reagent for testing comprises acetonitrile (chromatographic grade) and methanol (chromatographic grade), and ultrapure water (the conductivity meets the pharmacopoeia standard requirement) for a laboratory, and a polyoxypropylene ethylene oxide glycerol ether polyether standard;
preparing a standard solution, namely preparing a solution of polyoxypropylene ethylene oxide glycerol ether polyether and a diluent solution into 75.00 mug/ml solution, and filtering the solution through a water system filter membrane with the thickness of 0.22 mu m for later use;
preparing a sample solution, precisely measuring 0.25ml of the sample, diluting to a scale with acetonitrile in a 1ml measuring flask, shaking uniformly, centrifuging at 10000r for 5min, and filtering the supernatant with a water-based filter membrane of 0.22 mu m for later use;
step four, high performance liquid chromatography is used for testing, wherein the testing condition is a C-18 reversed phase chromatographic column with the size of 4.6X250 mm, and the mobile phase is as follows: a is methanol, B is ultrapure water for laboratory, and gradient elution is carried out; flow rate: 1.0ml/min; the detector adopts an electrospray detector, and uses polyoxypropylene ethylene oxide glyceryl ether polyether as a reference substance for quantification by an external standard method.
The biological medicine stock solution dissolved in acetonitrile is selected as a sample solution, and is detected by adopting a high performance liquid chromatography, and a C-18 reversed phase chromatographic column with the diameter of 4.6X250 mm is used; mobile phase A is methanol, and mobile phase B is laboratory ultrapure water for gradient elution.
Mobile phase a: methanol
Mobile phase B: laboratory ultrapure water
Elution gradient:
flow rate: 1.0ml/min
Column temperature: 30 DEG C
A detector: electrospray detector
Sample injection amount: 20 μl of
Run time: 50min
Preparing a test solution: precisely measuring 0.25ml of a sample, diluting with acetonitrile to a scale in a 1ml measuring flask, shaking uniformly, centrifuging at 10000r for 5min, and filtering the supernatant with a water-based filter membrane of 0.22 μm for later use;
polyoxypropylene ethylene oxide glycerol ether standard solution: preparing a solution of polyoxypropylene ethylene oxide glycerol ether polyether and a diluent solution into a solution of 75.00 mu g/ml, and filtering the solution through a water system filter membrane of 0.22 mu m for later use;
and (3) testing: sampling the sample solution to be tested and the standard reference solution respectively, collecting chromatograms, and calculating the content of polyoxypropylene ethylene oxide glycerol ether according to an external standard method by using the peak area;
the solvent is a mixed solution of a mobile phase A and a mobile phase B, the mobile phase A is methanol, the mobile phase B is ultrapure water, and the gradient elution is carried out.
Preferably, the methanol solution is of chromatographic grade; the conductivity of the ultrapure water meets the requirements of pharmacopoeia standards.
Preferably, the flow rate is 1.0ml/min.
Preferably, the column temperature is 30 ℃.
Preferably, the detector is an electrospray detector
Compared with the prior art, the invention has the beneficial effects that:
1. the newly established method for detecting the residual content of the polyoxypropylene ethylene oxide glycerol ether polyether has the advantages of high separation degree, accurate result and good precision.
2. The determination method is based on a separation mechanism of high performance liquid chromatography, adopts a novel universal detector, namely an electrospray detector CAD, has high detection sensitivity and good reproducibility, ensures the stability of a sample in the test process, and realizes the scientific determination of the polyoxypropylene ethylene oxide glyceryl ether polyether residue. The method is evaluated by the recovery rate and the relative standard deviation, the recovery rate is measured to be 95-106%, and the accuracy is good; the relative standard deviation of the parallel measurement of the samples is less than 2.0%, and the precision of the method is good.
Drawings
FIG. 1 is a chromatogram of polyoxypropylene ethylene oxide glyceryl ether polyether in a sample solution of the present invention;
FIG. 2 is a chromatogram of a detection limiting solution polyoxypropylene ethylene oxide glyceryl ether polyether of the present invention;
FIG. 3 is a quantitative limiting solution polyoxypropylene ethylene oxide glyceryl ether polyether chromatogram of the present invention;
FIG. 4 is a graph showing the linear relationship of the polyoxypropylene ethylene oxide glyceryl ether polyether standard control of example 3 of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples, which are not intended to limit the present invention, but are merely illustrative of the present invention. The experimental methods used in the following examples are not specifically described, but the experimental methods in which specific conditions are not specified in the examples are generally carried out under conventional conditions, and the materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.
Example 1
Detector and mobile phase selection
A detector: because the polyoxypropylene ethylene oxide glyceryl ether polyether has weaker ultraviolet absorption and an ultraviolet detector is not suitable, a novel universal detector CAD detector is selected, the novel universal detector CAD detector can be applied to non-volatile or semi-volatile compounds in a hot river based on a unique innovative testing principle, and has high detection sensitivity and good reproducibility.
Mobile phase a: methanol (chromatographic grade), mobile phase B: laboratory ultrapure water is used for gradient elution;
example 2
Conditions of main chromatography
Mobile phase a: methanol
Mobile phase B: laboratory ultrapure water
Elution gradient:
flow rate: 1.0ml/min
Column temperature: 30 DEG C
A detector: CAD detector
Sample injection amount: 20 μl of
Chromatographic column: reverse direction chromatography column C18 (4.6X1250 mm. Times.5 μm)
Preparing a test solution: precisely measuring 0.25ml of a sample, diluting with acetonitrile to a scale in a 1ml measuring flask, shaking uniformly, centrifuging at 10000r for 5min, and filtering the supernatant with a water-based filter membrane of 0.22 μm for later use;
preparing a standard reference substance solution: preparing a solution of polyoxypropylene ethylene oxide glycerol ether polyether and a diluent solution into a solution of 75.00 mu g/ml, and filtering the solution through a water system filter membrane of 0.22 mu m for later use;
sample injection is carried out on the sample solution to be tested and the standard reference solution respectively, a chromatogram is collected, and the content of the polyoxypropylene ethylene oxide glycerol ether polyether is calculated according to an external standard method by using the peak area.
Wherein, fig. 2 and 3 are the detection limit and quantitative limit chromatograms of the polyoxypropylene ethylene oxide glyceryl ether polyether, and fig. 1 is the chromatogram of the residual biological medicine stock solution containing the polyoxypropylene ethylene oxide glyceryl ether polyether. As can be seen in fig. 2 and 3, the retention time of the polyoxypropylene ethylene oxide glyceryl ether polyether was 43.65min. Determining the retention time of the polyoxypropylene ethylene oxide glyceryl ether polyether according to the peak position in a standard reference substance chromatogram, thereby determining the existence of the polyoxypropylene ethylene oxide glyceryl ether polyether in a biological medicine stock solution test sample; the content of polyoxypropylene ethylene oxide glyceryl ether polyether in the sample solution was calculated according to the peak area of fig. 1.
Example 3
Verification test
The chromatographic conditions of example 2 were examined in terms of system applicability, specificity, precision (reproducibility), precision (intermediate precision), linearity, limit of detection and limit of quantification, accuracy, etc., and the results were as follows:
(1) System applicability
And (3) continuously detecting the standard reference substance solution (about 75.00 mug/ml) for 6 times, calculating the RSD% value, and judging whether the applicability of the system meets the requirement. The results are shown in Table 1 below.
Table 1: system applicability results
The result shows that the RSD value of the polyoxypropylene ethylene oxide glyceryl ether polyether is 1.7% when the same reference substance solution is continuously detected for 6 times, and meets the requirement (the precision repeatability RSD% is less than or equal to 4.0 when the content of the components to be detected is 0.01%).
(2) Specialization of
And respectively taking a blank solution (acetonitrile), a sample solution (biological medicine stock solution is diluted by acetonitrile, and centrifuged), taking supernatant as the sample solution, and a polyoxypropylene ethylene oxide glycerol ether polyether reference substance solution, and sequentially carrying out sample injection analysis. As a result of detection, the retention time of the polyoxypropylene ethylene oxide glyceryl ether polyether is 43.671min, the peak separation effect in the sample solution is good (the separation degree is 2.0 & gt 1.5), the sample solution does not interfere with the control solution of the polyoxypropylene ethylene oxide glyceryl ether polyether, the retention time of the polyoxypropylene ethylene oxide glyceryl ether polyether is consistent, and the detection method is feasible and has strong specificity.
(3) Precision (repeatability)
6 parts of the sample to be tested is added with a standard solution (a certain amount of polyoxypropylene ethylene oxide glycerol ether polyether reference substance is added into the biological medicine stock solution) and then sample injection test is carried out, and the precision is calculated, and the result is shown in Table 2.
Table 2: precision (reproducibility) results
The result shows that the RSD value of the polyoxypropylene ethylene oxide glycerol ether polyether is 2.6% when 6 parts of the sample with the same concentration are tested under the same condition, and meets the requirement of precision (the RSD of the precision is less than or equal to 4.0% when the content of the component to be measured is 0.01%), so that the precision is good under the chromatographic condition.
(4) Precision (intermediate precision)
And 6 test sample adding solutions (a certain amount of polyoxypropylene ethylene oxide glycerol ether polyether reference substances are added into the biological medicine stock solution) are prepared by other 1 tester on different days, the sample is sequentially injected and tested, the precision is calculated, and the result is shown in Table 3.
Table 3: results of precision (intermediate precision)
The results of the intermediate precision of the 12 sample concentrations are shown in Table 4.
Table 4: results of intermediate precision of 12 parts solution
The result shows that the recovery rate RSD of 12 parts of test sample adding standard solution of different testers is 2.1 percent and is not more than 8.0 percent, and the intermediate precision meets the requirement (when the content of the component to be measured is 0.01 percent, the precision reproducibility RSD percent is less than or equal to 8.0 percent). And the recovery rate of the 12 parts of sample adding standard solution is 97-103%, and is 85-110%, and the recovery rate meets the requirements.
(5) Linearity of
Preparing a polyoxypropylene ethylene oxide glycerol ether polyether standard reference substance solution, diluting the solution into a series of solutions with the concentration of 19.936 mug/ml, 37.474 mug/ml, 74.948 mug/ml, 112.421 mug/ml and 149.895 mug/ml, sequentially injecting samples,
a linear plot of concentration versus peak area was measured as shown in table 5.
Table 5: linear results
Conclusion: the polyoxypropylene ethylene oxide glyceryl ether polyether has good linear relation within the concentration range of 20-150 mug/ml, the correlation coefficient is 0.991 and is not less than 0.990, and the linearity and the range meet the requirements.
(6) Limit of detection and limit of quantification
And diluting the polyoxypropylene ethylene oxide glycerol ether polyether reference substance solution into a series of concentrations, and sequentially measuring to obtain the polyoxypropylene ethylene oxide glycerol ether polyether with the detection limit of 10.041 mug/ml and the quantitative limit of 19.935 mug/ml, wherein the signal to noise ratio is not less than 10, and the requirements are met.
(7) Accuracy of
Preparing low-concentration labeled sample solution, medium-concentration labeled sample solution and high-concentration labeled sample solution, respectively 3 parts, sequentially detecting 9 parts of the solutions, and calculating recovery rate as shown in table 6.
Table 6: accuracy results
The result shows that the RSD value of the polyoxypropylene ethylene oxide glyceryl ether polyether in 3 parts of the low-concentration standard test sample solution, the medium-concentration standard test sample solution and the high-concentration standard test sample solution respectively is 1.9 percent, meets the requirement of accuracy (the accuracy RSD is less than or equal to 4.0 when the content of the component to be measured is 0.01 percent), and the recovery rate of the 9 parts of solution is 96-101 percent, meets the requirement of 85-110 percent, so that the accuracy is good under the chromatographic condition.
And (3) detecting a sample:
3 batches of samples were tested using the above-identified method and the results are shown in Table 7.
The results show that the method has good specificity and can rapidly and accurately detect the amount of the residual polyoxypropylene ethylene oxide glyceryl ether polyether in the biological medicine stock solution.
Example 4
Referring to the procedure of example 2, the column flow rate was varied and the results are shown in Table 8.
Table 8: investigation results of chromatographic column flow velocity
The result shows that the column flow rate is changed within the range of 0.80-1.35 ml/min, the influence on the content measurement of the polyoxypropylene ethylene oxide glyceryl ether polyether is small, the peak value of the polyoxypropylene ethylene oxide glyceryl ether polyether is good, and the accurate quantification can be realized.
Example 5
Referring to the procedure of example 2, the column temperature was changed and the results are shown in Table 9.
Table 9: investigation result of column temperature
The result shows that the column temperature is changed within the range of 25-40 ℃, the influence on the content measurement of the polyoxypropylene ethylene oxide glyceryl ether polyether is small, the peak type of the polyoxypropylene ethylene oxide glyceryl ether polyether is good, and the accurate quantification can be realized.
The foregoing examples illustrate only a few embodiments of the invention and are described in detail herein without thereby limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (5)
1. A quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in the production of biological drugs is characterized by comprising the following steps:
the biological medicine stock solution dissolved in acetonitrile is selected as a sample solution, and is detected by adopting a high performance liquid chromatography, and a C-18 reversed phase chromatographic column with the diameter of 4.6X250 mm is used; mobile phase A is methanol, and mobile phase B is laboratory ultrapure water for gradient elution.
Mobile phase a: methanol
Mobile phase B: laboratory ultrapure water
Elution gradient:
flow rate: 1.0ml/min
Column temperature: 30 DEG C
A detector: electrospray detector
Sample injection amount: 20 μl of
Run time: 50min
Preparing a test solution: precisely measuring 0.25ml of a sample, diluting with acetonitrile to a scale in a 1ml measuring flask, shaking uniformly, centrifuging at 10000r for 5min, and filtering the supernatant with a water-based filter membrane of 0.22 μm for later use;
polyoxypropylene ethylene oxide glycerol ether standard solution: preparing a solution of polyoxypropylene ethylene oxide glycerol ether polyether and a diluent solution into a solution of 75.00 mu g/ml, and filtering the solution through a water system filter membrane of 0.22 mu m for later use;
and (3) testing: sampling the sample solution to be tested and the standard reference solution respectively, collecting chromatograms, and calculating the content of polyoxypropylene ethylene oxide glycerol ether according to an external standard method by using the peak area;
the solvent is a mixed solution of a mobile phase A and a mobile phase B, the mobile phase A is methanol, the mobile phase B is ultrapure water, and the gradient elution is carried out.
2. The method for quantitatively detecting polyoxypropylene ethylene oxide glycerol ether residue in the production of biological drugs according to claim 1, which is characterized in that: the methanol solution is of chromatographic grade; the conductivity of the ultrapure water meets the standard requirements of pharmacopoeia.
3. The method for quantitatively detecting polyoxypropylene ethylene oxide glycerol ether residue in the production of biological drugs according to claim 1, which is characterized in that: the flow rate was 1.0ml/min.
4. The method for quantitatively detecting polyoxypropylene ethylene oxide glycerol ether residue in the production of biological drugs according to claim 1, which is characterized in that: the column temperature was 30 ℃.
5. The method for quantitatively detecting polyoxypropylene ethylene oxide glycerol ether residue in the production of biological drugs according to claim 1, which is characterized in that: the detector is an electrospray detector.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410063462.1A CN117871752A (en) | 2024-01-16 | 2024-01-16 | Quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in biological medicine stock solution |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410063462.1A CN117871752A (en) | 2024-01-16 | 2024-01-16 | Quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in biological medicine stock solution |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117871752A true CN117871752A (en) | 2024-04-12 |
Family
ID=90584386
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410063462.1A Pending CN117871752A (en) | 2024-01-16 | 2024-01-16 | Quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in biological medicine stock solution |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117871752A (en) |
-
2024
- 2024-01-16 CN CN202410063462.1A patent/CN117871752A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | Elimination of matrix effects for headspace solid-phase microextraction of important volatile compounds in red wine using a novel coating | |
| CN108152425B (en) | Method for detecting lignanoids in sesame oil by high performance liquid chromatography | |
| CN104215705A (en) | Method for detecting residual amount of organic-chlorine pesticide in grain | |
| CN113341038A (en) | Method for measuring content of emamectin benzoate | |
| CN101122589A (en) | Urea and its impurity high performance liquid chromatography analysis method | |
| CN113533548A (en) | Method for detecting 1-vinyl imidazole in chemical products | |
| CN117871752A (en) | Quantitative detection method for polyoxypropylene ethylene oxide glycerol ether residues in biological medicine stock solution | |
| CN113607836B (en) | Analysis method for content of indoxacarb key intermediate | |
| Murugan et al. | A review on method development and validation by using HPLC | |
| Matisová et al. | Solid‐phase microextraction of volatile polar compounds in water | |
| CN108663457A (en) | Utilize the method for residual acrylic acid in HS-GC external standard method acrylate copolymers | |
| CN108445106A (en) | The remaining detection method of antifoaming agent polypropylene glycol in a kind of drug | |
| CN109298092B (en) | HPLC method for detecting content of methylsulfonyl chloride in industrial waste liquid | |
| Wang et al. | Rapid method for the determination of the stable oxygen isotope ratio of water in alcoholic beverages | |
| CN108181393B (en) | Method for detecting hydroxyethyl hexahydro-s-triazine in plastic product | |
| CN110161134B (en) | Method for detecting nucleic acid solid sample methylamine and methylamine salt solvent residue | |
| CN116930368B (en) | Detection method of settop alcohol isomer | |
| CN114200067B (en) | High performance liquid chromatography analysis method for 6-bromo-3-hydroxy pyrazine-2-carboxamide and impurities | |
| CN112305100B (en) | Method for detecting content of genotoxic impurity benzyl bromide in medicine | |
| CN110687226B (en) | Method for measuring content of 2-chloropropionic acid in iopamidol | |
| CN109507327B (en) | Quantitative determination of TNT content by GC-AED independent calibration curve method (CIC method) | |
| CN115616120B (en) | Method for simultaneously measuring contents of 11 feeding organic acids in organic acidifier | |
| CN109521120B (en) | Quantitative determination of DNTF content by GC-AED independent calibration curve method (CIC method) | |
| CN101769872B (en) | Detection method of cordycepic acid content in cordyceps product | |
| CN106885855A (en) | A kind of Gas Chromatographic Determination of 1,1,1 3 chloronitroethane |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |