CN117571846A - Method for detecting content of minoxidil in external solution - Google Patents
Method for detecting content of minoxidil in external solution Download PDFInfo
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- CN117571846A CN117571846A CN202311291234.1A CN202311291234A CN117571846A CN 117571846 A CN117571846 A CN 117571846A CN 202311291234 A CN202311291234 A CN 202311291234A CN 117571846 A CN117571846 A CN 117571846A
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- minoxidil
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- ZFMITUMMTDLWHR-UHFFFAOYSA-N Minoxidil Chemical compound NC1=[N+]([O-])C(N)=CC(N2CCCCC2)=N1 ZFMITUMMTDLWHR-UHFFFAOYSA-N 0.000 title claims abstract description 44
- 229960003632 minoxidil Drugs 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 39
- 239000000243 solution Substances 0.000 claims abstract description 43
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 30
- 238000001514 detection method Methods 0.000 claims abstract description 23
- 239000003085 diluting agent Substances 0.000 claims abstract description 23
- 239000000872 buffer Substances 0.000 claims abstract description 11
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims abstract description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000013558 reference substance Substances 0.000 claims description 14
- 239000012488 sample solution Substances 0.000 claims description 13
- 239000011550 stock solution Substances 0.000 claims description 12
- 238000005303 weighing Methods 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 8
- 239000008213 purified water Substances 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 6
- REFMEZARFCPESH-UHFFFAOYSA-M sodium;heptane-1-sulfonate Chemical compound [Na+].CCCCCCCS([O-])(=O)=O REFMEZARFCPESH-UHFFFAOYSA-M 0.000 claims description 4
- 238000010812 external standard method Methods 0.000 claims description 3
- 239000012467 final product Substances 0.000 claims description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 2
- 239000007853 buffer solution Substances 0.000 claims description 2
- 238000012937 correction Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 claims description 2
- 238000012360 testing method Methods 0.000 abstract description 24
- 238000011084 recovery Methods 0.000 abstract description 7
- 238000013112 stability test Methods 0.000 abstract description 5
- 239000003814 drug Substances 0.000 abstract description 4
- 239000002253 acid Substances 0.000 abstract description 3
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 238000012795 verification Methods 0.000 abstract description 3
- 238000004458 analytical method Methods 0.000 abstract description 2
- 239000011259 mixed solution Substances 0.000 abstract 1
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 abstract 1
- 239000000741 silica gel Substances 0.000 abstract 1
- 229910002027 silica gel Inorganic materials 0.000 abstract 1
- 238000005457 optimization Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 3
- 239000012085 test solution Substances 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000012088 reference solution Substances 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 201000004384 Alopecia Diseases 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 231100000360 alopecia Toxicity 0.000 description 1
- 208000004631 alopecia areata Diseases 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000003779 hair growth Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000010829 isocratic elution Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 229940046530 minoxidil topical solution Drugs 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 239000000810 peripheral vasodilating agent Substances 0.000 description 1
- 229960002116 peripheral vasodilator Drugs 0.000 description 1
- 238000003918 potentiometric titration Methods 0.000 description 1
- 229960004063 propylene glycol Drugs 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- 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
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a method for detecting the content of minoxidil in an external solution, which belongs to the field of medicine analysis and detection, and comprises the following steps: adopting octadecylsilane chemically bonded silica gel chromatographic column, taking mixed solution of methanol-buffer solution-phosphoric acid as mobile phase, wherein the flow rate is 0.9-1.1mL/min, the column temperature is 25-35 ℃, and the detection wavelength of the sample is 225-235 nm. The method of the invention uses the mobile phase without adding acid as the diluent, so that the solution system is more stable, the detection repeatability is good, the sensitivity is high, and the detection result is more accurate. The method of the invention carries out methodology verification tests as follows: the method is verified by a linearity and range test, a recovery rate test, a repeatability test, a precision test and a stability test, and the result shows that the method is accurate and reliable.
Description
Technical Field
The invention belongs to the field of medicine analysis and detection, and particularly relates to a method for detecting the content of minoxidil in an external solution, which adopts a high performance liquid chromatograph to directly detect.
Background
Minoxidil is a peripheral vasodilator which can be used for the treatment of hypertension, but it has been found in clinical use that this drug also has the effect of stimulating hair growth and can be used for the treatment of seborrheic alopecia and alopecia areata when used for a long period of time. The 2020 edition of Chinese pharmacopoeia prescribes that the minoxidil content is measured by a potentiometric titration method, but the minoxidil external solution is a liquid preparation prepared from minoxidil, an antioxidant, a metal ion chelating agent, a polyalcohol solvent and purified water. In order to further perfect the quality control of the minoxidil external solution and ensure the quality of medicines, a method for detecting the content of the minoxidil external solution is very necessary.
Disclosure of Invention
The invention aims to: the invention aims to solve the technical problems of high detection cost, unstable detection solution system and inaccurate detection result of the existing detection method for the content of the main component in the minoxidil external solution, and provides a method for detecting the content of the minoxidil external solution with low detection cost, stable detection solution system and accurate detection result.
The technical scheme is as follows: a method for detecting the content of minoxidil in an external solution comprises the following steps:
step one, setting chromatographic conditions:
1. chromatographic column: agilent ZORBAX Eclipse Plus C18 (4.6X105 mm,3.5 μm);
2. column temperature: 25-35 ℃;
3. flow rate: 0.9 to 1.1ml per minute;
4. detection wavelength: 225nm-235 nm;
5. mobile phase: methanol: buffer 1: phosphoric acid = 430:570:1 to 470:530:1, a step of;
6. a diluent: methanol: buffer 2 = 450:550;
step two, preparing a reference substance solution:
a. weighing minoxidil reference substance about 20mg, precisely weighing, placing into 50ml measuring flask, dissolving with diluent, fixing volume to scale, shaking, and collecting as stock solution;
b. precisely measuring 1.0ml of the stock solution, placing in a 10ml measuring flask, fixing volume to scale with diluent, and shaking;
step three, preparing a sample solution:
a. precisely measuring 1.0ml of sample solution (about equivalent to 20mg containing minoxidil), placing in a 50ml measuring flask, adding diluent to scale, and shaking;
b. precisely measuring 1.0ml of the solution, placing in a 10ml measuring flask, fixing the volume to scale with diluent, and shaking uniformly to obtain the final product;
step four, measuring and calculating:
and (3) respectively precisely sucking 10 mu l of the reference substance solution prepared in the second step and 10 mu l of the sample solution prepared in the third step, injecting into a liquid chromatograph, measuring, recording and calculating by peak area according to an external standard method to obtain the content of minoxidil in the external solution by using the correction factor RSD of minoxidil in the reference substance solution to be less than or equal to 2.0 percent.
As an optimization: the column temperature was 30 ℃.
As an optimization: the flow rate was 1.0mL per minute.
As an optimization: the detection wavelength is 230nm.
As an optimization: methanol in the mobile phase: buffer 1: phosphoric acid = 450:550:1.
as an optimization: the preparation of buffer 1 in the mobile phase is as follows: 2g of 1-heptanesulfonic acid sodium salt was weighed into 550ml of purified water, and stirred to be completely dissolved, and filtered through a 0.45 μm aqueous membrane.
As an optimization: the preparation of the buffer solution 2 in the diluent is as follows: accurately weighing 2g of 1-heptane sodium sulfonate, placing into 550ml of purified water, stirring to dissolve completely, and filtering with 0.45 μm water system membrane.
The beneficial effects are that: the method of the invention uses the mobile phase without adding acid as the diluent, so that the solution system is more stable, the detection repeatability is good, the sensitivity is high, and the detection result is more accurate. The method of the invention carries out methodology verification tests as follows: the method is verified by a linearity and range test, a recovery rate test, a repeatability test, a precision test and a stability test, and the result shows that the method is accurate and reliable.
Drawings
FIG. 1 is a chromatogram of minoxidil control solution measured according to the minoxidil external solution content detection method provided by the invention;
FIG. 2 is a chromatogram of a test solution measured by the minoxidil external solution content detection method provided by the invention;
fig. 3 is a standard graph of the minoxidil external solution content detection method provided by the invention.
Detailed Description
The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent. It will be apparent that the described embodiments are some, but not all, embodiments of the invention.
The specific techniques or conditions are not identified in the examples and are described in the literature or are carried out in accordance with the specifications of the product. The equipment and other manufacturers are not noted, and the equipment and the other manufacturers are conventional products which can be purchased through regular channels. The methods are conventional methods unless otherwise specified, and the starting materials are commercially available from the public sources unless otherwise specified. The specific techniques or conditions are not identified in the examples and are described in the literature in this field or are carried out in accordance with the product specifications.
Example 1
A method for detecting the content of minoxidil in an external solution comprises the following steps:
1. instrument: agilent 1260 high performance liquid chromatograph; meltrel-tolidox XP105 electronic balance; agilent column Agilent ZORBAX Eclipse Plus C (4.6X150 mm,3.5 μm).
2. Chromatographic conditions: the chromatographic column was Agilent ZORBAX Eclipse Plus C (4.6X105 mm,3.5 μm), and isocratic elution was performed with methanol-buffer (2 g of 1-heptanesulfonic acid sodium salt was accurately weighed and placed in 550ml of purified water, stirred to dissolve completely, and filtered through a 0.45 μm aqueous membrane) -phosphoric acid (450:550:1) as the mobile phase; methanol is needle washing liquid; the flow rate is 1.0ml per minute; the detection wavelength is 230nm; column temperature was 30 ℃.
3. A diluent: methanol-buffer (2 g of 1-heptanesulfonic acid sodium salt was accurately weighed and placed in 550ml of purified water, and stirred to be completely dissolved, and filtered through a 0.45 μm aqueous membrane) (450:550).
4. Preparing a reference substance solution: weighing minoxidil reference substance about 20mg, precisely weighing, placing into 50ml measuring flask, dissolving with diluent, fixing volume to scale, shaking, and collecting as stock solution. Precisely measuring 1.0ml of the stock solution, placing into a 10ml measuring flask, fixing volume to scale with diluent, and shaking.
5. Sample solution preparation: 1.0ml of the sample solution (approximately equivalent to 20mg containing minoxidil) was precisely measured, placed in a 50ml measuring flask, diluted to scale and shaken well. Precisely measuring 1.0ml of the solution, placing in a 10ml measuring flask, fixing the volume to scale with diluent, and shaking to obtain the final product.
6. The measuring step comprises the following steps: respectively precisely sucking 10 μl of the reference solution and the sample solution, respectively, injecting into a liquid chromatograph, measuring, and calculating with peak area according to external standard method to obtain the content of minoxidil topical solution.
The prior method is compared with the method of the invention: the method uses the mobile phase without adding acid as the diluent, so that the solution system is more stable, the detection repeatability is good, the sensitivity is high, and the detection result is more accurate.
The related performance test experiment and experimental result in the invention are as follows:
1. linearity and Range test
The minoxidil reference substance 20mg (content 99.7%) was precisely weighed, placed in a 50ml measuring flask, dissolved with a diluent and fixed to a scale, and shaken well as a linear stock solution. A proper amount of the linear stock solution was precisely aspirated, diluted with a diluent to prepare reference solutions having concentrations of about 20.0. Mu.g/ml, 32.0. Mu.g/ml, 40.0. Mu.g/ml, 50.0. Mu.g/ml, and 60.0. Mu.g/ml, respectively, and injected into a liquid chromatograph, and the results were determined according to the above chromatographic conditions and are shown in Table 1. A standard curve was plotted with minoxidil concentration (μg/ml) as the abscissa and the corresponding peak area as the ordinate (see FIG. 3). The results show that minoxidil has a good linear relationship in the range, and the regression equation is y=63.431x+23.631, and r=1.0000.
Table 1 results of linear investigation
2. Precision test
The minoxidil reference substance solution was continuously sampled 6 times by precise suction, the sample feeding amount was 10 μl, the chromatogram was recorded, the minoxidil peak area was measured, and the Relative Standard Deviation (RSD) was calculated, and the results are shown in table 2, RSD (n=6) was 0.08%, showing that the method was excellent in precision.
TABLE 2 results of precision test
3. Repeatability test
6 parts of samples with the same batch number (M1S 220501) are taken, the content of 6 parts of samples is measured according to the preparation and chromatographic conditions of the sample solution, the result is shown in a table 3, the average content is 101.9%, and the RSD is 0.27%, which indicates that the method has good repeatability.
TABLE 3 repeatability test results
4. Stability test
Precisely sucking the control solution and the sample solution, and respectively measuring peak areas of minoxidil at 0, 4, 7, 20, 34 and 56 hours after preparation at room temperature, wherein 10 μl of each sample is injected, and the peak area RSD of the minoxidil control solution within 56 hours is measured to be 0.79%; the peak area RSD of the test solution was 0.31%. The test results are shown in Table 4, which shows that minoxidil control solution and test solution were stable at room temperature for 56 hours.
TABLE 4 stability test results
5. Recovery test
Weighing 15ml of absolute ethyl alcohol and 25ml to 50ml of 1, 2-propylene glycol into a measuring flask, adding purified water for dilution, fixing the volume to a scale, and shaking uniformly to obtain an empty auxiliary material stock solution. Precisely weighing minoxidil reference substance 20mg (content 99.7%), placing into 50ml measuring flask, dissolving with diluent, fixing volume to scale, shaking, and taking as reference substance stock solution. 1ml of blank auxiliary material stock solution is measured and placed in a 10ml volumetric flask, 0.8ml, 1.0ml and 1.2ml of reference material stock solution are added respectively, and diluted and fixed volume is carried out by diluent until the scale is reached to prepare the accurate sample solutions with the concentration of about 320 mug/ml, 400 mug/ml and 500 mug/ml. 3 parts of test sample solutions were prepared for each concentration, and the recovery rate was measured by the content measurement method, and RSD (n=9) was found to be the result shown in table 5, indicating that the method has a good recovery rate.
TABLE 5 recovery test results
In summary, the method of the invention performs a methodological verification test, and specifically comprises the following steps: the method is verified by a linearity and range test, a recovery rate test, a repeatability test, a precision test and a stability test, and the result shows that the method is accurate and reliable.
Claims (7)
1. A method for detecting the content of minoxidil in an external solution is characterized by comprising the following steps: the method comprises the following steps:
step one, setting chromatographic conditions:
a) Chromatographic column: agilent ZORBAX Eclipse Plus C18 (4.6X105 mm,3.5 μm);
b) Column temperature: 25-35 ℃;
c) Flow rate: 0.9 to 1.1ml per minute;
d) Detecting wavelength: 225nm-235 nm;
e) Mobile phase: methanol: buffer 1: phosphoric acid = 430:570:1 to 470:530:1, a step of;
f) Diluting agent: methanol: buffer 2 = 450:550;
step two, preparing a reference substance solution:
a. weighing minoxidil reference substance about 20mg, precisely weighing, placing into 50ml measuring flask, dissolving with diluent, fixing volume to scale, shaking, and collecting as stock solution;
b. precisely measuring 1.0ml of the stock solution, placing in a 10ml measuring flask, fixing volume to scale with diluent, and shaking;
step three, preparing a sample solution:
(a) Precisely measuring 1.0ml of sample solution (about equivalent to 20mg containing minoxidil), placing in a 50ml measuring flask, adding diluent to scale, and shaking;
(b) Precisely weighing 1.0ml of the solution, placing in a 10ml measuring flask, fixing the volume to scale with diluent, and shaking uniformly to obtain the final product;
step four, measuring and calculating:
and (3) respectively precisely sucking 10 mu l of the reference substance solution prepared in the second step and 10 mu l of the sample solution prepared in the third step, injecting into a liquid chromatograph, measuring, recording and calculating by peak area according to an external standard method to obtain the content of minoxidil in the external solution by using the correction factor RSD of minoxidil in the reference substance solution to be less than or equal to 2.0 percent.
2. The method for detecting the content of minoxidil in an external solution according to claim 1, wherein: the column temperature was 30 ℃.
3. The method for detecting the content of minoxidil in an external solution according to claim 1, wherein: the flow rate was 1.0mL per minute.
4. The method for detecting the content of minoxidil in an external solution according to claim 1, wherein: the detection wavelength is 230nm.
5. The method for detecting the content of minoxidil in an external solution according to claim 1, wherein: methanol in the mobile phase: buffer 1: phosphoric acid = 450:550:1.
6. the method for detecting the content of minoxidil in an external solution according to claim 1, wherein: the preparation of buffer 1 in the mobile phase is as follows: 2g of 1-heptanesulfonic acid sodium salt was weighed into 550ml of purified water, and stirred to be completely dissolved, and filtered through a 0.45 μm aqueous membrane.
7. The method for detecting the content of minoxidil in an external solution according to claim 1, wherein: the preparation of the buffer solution 2 in the diluent is as follows: accurately weighing 2g of 1-heptane sodium sulfonate, placing into 550ml of purified water, stirring to dissolve completely, and filtering with 0.45 μm water system membrane.
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