CN115219613B - Method for detecting azelaic acid and potassium azelate in cosmetics - Google Patents
Method for detecting azelaic acid and potassium azelate in cosmetics Download PDFInfo
- Publication number
- CN115219613B CN115219613B CN202210682555.3A CN202210682555A CN115219613B CN 115219613 B CN115219613 B CN 115219613B CN 202210682555 A CN202210682555 A CN 202210682555A CN 115219613 B CN115219613 B CN 115219613B
- Authority
- CN
- China
- Prior art keywords
- solution
- azelaic acid
- potassium
- standard
- sample
- 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.)
- Active
Links
- BDJRBEYXGGNYIS-UHFFFAOYSA-N nonanedioic acid Chemical compound OC(=O)CCCCCCCC(O)=O BDJRBEYXGGNYIS-UHFFFAOYSA-N 0.000 title claims abstract description 159
- RTMGQVSGXYZVTE-UHFFFAOYSA-L dipotassium;nonanedioate Chemical compound [K+].[K+].[O-]C(=O)CCCCCCCC([O-])=O RTMGQVSGXYZVTE-UHFFFAOYSA-L 0.000 title claims abstract description 54
- 238000000034 method Methods 0.000 title claims abstract description 38
- 239000002537 cosmetic Substances 0.000 title claims abstract description 33
- 238000001514 detection method Methods 0.000 claims abstract description 27
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 102
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 74
- 239000000243 solution Substances 0.000 claims description 64
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 18
- 239000011550 stock solution Substances 0.000 claims description 18
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 16
- 238000005303 weighing Methods 0.000 claims description 14
- 239000013582 standard series solution Substances 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 10
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 9
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 8
- 238000010828 elution Methods 0.000 claims description 8
- 238000009210 therapy by ultrasound Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 7
- 239000008055 phosphate buffer solution Substances 0.000 claims description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 6
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 6
- 239000012498 ultrapure water Substances 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 238000004090 dissolution Methods 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 3
- 238000000967 suction filtration Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 abstract description 7
- 238000001212 derivatisation Methods 0.000 abstract description 4
- 238000000605 extraction Methods 0.000 abstract description 4
- 239000002904 solvent Substances 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 35
- 238000011084 recovery Methods 0.000 description 15
- 239000012488 sample solution Substances 0.000 description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 239000007864 aqueous solution Substances 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 238000005259 measurement Methods 0.000 description 8
- -1 potassium nonyldiglycolate Chemical compound 0.000 description 8
- 239000011159 matrix material Substances 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- 230000004044 response Effects 0.000 description 6
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 4
- 238000002372 labelling Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- UDOPWDMIGDFPBW-UHFFFAOYSA-N nonanedioic acid;potassium Chemical compound [K].OC(=O)CCCCCCCC(O)=O UDOPWDMIGDFPBW-UHFFFAOYSA-N 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 208000002874 Acne Vulgaris Diseases 0.000 description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 3
- 206010000496 acne Diseases 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 239000006071 cream Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000004817 gas chromatography Methods 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 238000012417 linear regression Methods 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 108010008488 Glycylglycine Proteins 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000012496 blank sample Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- YMAWOPBAYDPSLA-UHFFFAOYSA-N glycylglycine Chemical compound [NH3+]CC(=O)NCC([O-])=O YMAWOPBAYDPSLA-UHFFFAOYSA-N 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 230000002087 whitening effect Effects 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 229920000715 Mucilage Polymers 0.000 description 1
- 206010029098 Neoplasm skin Diseases 0.000 description 1
- 208000000453 Skin Neoplasms Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 239000000058 anti acne agent Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 229940124340 antiacne agent Drugs 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 230000009759 skin aging Effects 0.000 description 1
- 208000017520 skin disease Diseases 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 230000000699 topical effect Effects 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
-
- 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
Abstract
The invention discloses a simultaneous detection method of azelaic acid and potassium azelate in cosmetics. By adopting the method, the sample can be directly measured by a high performance liquid chromatograph without derivatization after being extracted by an extraction solvent, the method is simple and convenient, easy to operate, strong in universality and good in reproducibility, and is more suitable for popularization and application of the method between laboratories.
Description
Technical Field
The invention relates to the field of detection and analysis of cosmetics, in particular to a detection method of azelaic acid and potassium azelate in cosmetics.
Background
Azelaic acid medicine was developed by scheing AG, germany, first marketed in france in 1989 as a new anti-acne agent. Topical azelaic acid pharmaceutical formulations currently come in 3 classes: gels, creams and mucilage. In recent years, azelaic acid has remarkable effects on the protection of skin and the treatment of skin diseases such as acne treatment, skin aging improvement, skin inflammation improvement, skin tumor improvement and the like. It can effectively inhibit acne bacillus on skin and block fatty acid generation, and can also slow down keratinocyte growth, block melanin formation, prevent speckle formation and reduce melanin pigmentation. Azelaic acid has been widely used in cosmetics for whitening and removing acne in recent years due to its whitening and stain removing effects.
Potassium azelate is a novel azelate derivative which solves the difficulties in many formulations of azelaic acid, has better water solubility than azelaic acid, has the characteristics of small dosage and strong anti-inflammatory effect, and is therefore popular with cosmetic manufacturers in recent years as an alternative product of azelaic acid.
At present, in 2015 edition of "cosmetic health Specification" in China, no detection method and limit value of azelaic acid and potassium azelate are specified, and the national standard plan is numbered 201909-48-T-469: the detection of azelaic acid in cosmetics, gas chromatography, is under approval, but the method is a derivatization method followed by gas chromatography measurement, and is relatively cumbersome and difficult to operate. The method is not suitable for the determination of azelaic acid in cosmetics with complex matrixes, and is easy to generate matrix interference. And the method only measures azelaic acid in cosmetics, and does not carry out the method for measuring the content of the potassium azelate. Literature reports on the method for measuring the content of the azelaic acid at home and abroad are mainly focused on the fields of medicines, biological products and the like, and the adopted analysis technology mainly comprises high performance liquid chromatography, gas chromatography, capillary electrophoresis chromatography and the like. The method has the advantages of complicated operation steps and poor reproducibility because of sample injection analysis after derivatization of azelaic acid. The method for measuring the content of the potassium nonyldiglycolate in the cosmetics is not reported in the literature at present.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a method for simultaneously detecting azelaic acid and potassium azelate in cosmetics, which is simple and convenient, easy to operate, strong in universality and good in reproducibility, and is suitable for popularization and application of methods among laboratories.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
provides a method for detecting azelaic acid and potassium azelate in cosmetics, which comprises the following steps:
step one, preparing standard series solutions: precisely weighing azelaic acid and potassium azelate standard substances in a measuring flask respectively, dissolving the azelaic acid and the potassium azelate standard substances in methanol, fixing the volume to a scale, shaking the solution uniformly to serve as an azelaic acid and potassium azelate standard stock solution, accurately transferring the standard stock solution, fixing the volume by methanol, and preparing to obtain a standard series of solutions;
step two, preparing a solution to be measured: precisely weighing a sample to be measured, adding a sodium hydroxide aqueous solution-methanol solution, performing vortex treatment, adding the sodium hydroxide aqueous solution-methanol solution to fix the volume, performing ultrasonic treatment at 50-60 ℃ for 60-90min, and centrifuging for 5-20min to obtain a solution to be measured;
and thirdly, connecting the standard series solution prepared in the first step and the solution to be detected prepared in the second step to a high performance liquid chromatography system for detection.
Further, the concentrations of the standard series of solutions are respectively: 10.0. Mu.g/mL, 50.0. Mu.g/mL, 100.0. Mu.g/mL, 200.0. Mu.g/mL, 500. Mu.g/mL.
Further, the specific preparation method of the solution to be tested comprises the following steps: weighing 0.20g of a sample to be measured, placing the sample into a 50mL colorimetric tube with a plug, adding 45mL of sodium hydroxide aqueous solution-methanol solution, performing vortex treatment for 1min, adding the sodium hydroxide aqueous solution-methanol solution to fix the volume to 50mL, performing ultrasonic treatment at 60 ℃ for 90min, and centrifuging at 10000rpm for 5min to obtain the solution to be measured.
Further, high performance liquid chromatography conditions:
chromatographic column: B362V1 Swedish SVEAC8 column, 250mm×4.6mm×5 μm; or an equivalent chromatographic column;
column temperature: 30 ℃;
mobile phase: solution A is phosphate buffer solution; solution B is acetonitrile; gradient elution;
flow rate: 1.00mL/min;
sample injection amount: 10. Mu.L;
detection wavelength: 210nm.
Further, the gradient elution procedure was:
further, the preparation method of the phosphate buffer solution comprises the following steps: precisely weighing 2.0-2.5g of potassium dihydrogen phosphate, adding 1000mL of ultrapure water for dissolution, mixing uniformly, adding phosphoric acid for regulating the pH to 3.1, and carrying out suction filtration through a 0.45 mu m filter membrane.
Further, the mass concentration of the phosphoric acid is 85%.
Compared with the prior art, the invention has the following technical effects:
1) The extraction method can rapidly, completely and accurately extract azelaic acid and potassium azelate in cosmetics; the method for simultaneously measuring azelaic acid and the derivative of the azelaic acid, namely the potassium azelaic acid and the derivative of the azelaic acid, potassium azelaic acid in the cosmetics by adopting the high performance liquid chromatography is simple, convenient, efficient and easy to implement. By adopting the method, the sample can be directly measured by a high performance liquid chromatograph without derivatization after being extracted by an extraction solvent, the method is simple and convenient, easy to operate, strong in universality and good in reproducibility, and is more suitable for popularization and application of the method between laboratories.
2) By adopting the detection method of the invention, the recovery rate of azelaic acid and potassium azelate in the cosmetic sample can meet the requirement.
3) The detection method is adopted to detect the cosmetic sample actually containing azelaic acid and azelaic acid diglycemic acid potassium, and the detection result is consistent with the actual content of the sample; the method is accurate in measurement result in practical application.
4) The detection method can make up for the blank of the method of azelaic acid and potassium azelate in the existing cosmetics, the measurement result is close to the true value, and the detection capability of the detection method is correspondingly improved, so that the detection method has wide application prospect and good market potential.
Drawings
FIG. 1 is a high performance liquid chromatogram of a standard solution of azelaic acid and potassium azelate in an embodiment of the invention; wherein, 1-azelaic acid, 2-nonanedioyl diglycine potassium;
FIG. 2 is a high performance liquid chromatogram of a cosmetic sample solution containing azelaic acid according to one embodiment of the invention;
FIG. 3 is a high performance liquid chromatogram of a cosmetic sample solution containing potassium nonyldiglycolate in accordance with one embodiment of the present invention.
Detailed Description
The invention is further described below with reference to the drawings and specific examples, which are not intended to be limiting. It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other.
The invention provides a method for detecting azelaic acid and potassium azelate in cosmetics, which comprises the following steps:
1. reagents and materials
The reagents used in the invention are all analytically pure or below, and the water is primary water specified in GB/T6682 unless specified otherwise.
1.1 standard, see Table A.
Table A Standard substance information Table
1.2 acetonitrile, chromatographically pure.
1.3 methanol, chromatographically pure.
1.4 phosphoric acid (85%), superior purity.
1.5 sodium hydroxide, analytical grade
1.6 Potassium dihydrogen phosphate, analytical grade
1.7 aqueous sodium hydroxide solution (60 mmol/L): 2.4g of sodium hydroxide (1.5) is weighed, 1000mL of water is added, and the mixture is dissolved and mixed uniformly.
1.8 aqueous sodium hydroxide solution (60 mmol/L) -methanol solution (1+1): 100mL of an aqueous sodium hydroxide solution (60 mmol/L) (1.7) was added with 100mL of methanol (1.3), and the mixture was homogenized.
1.9 phosphate buffer: accurately weighing 2.0401g of potassium dihydrogen phosphate (1.6), adding 1000mL of ultrapure water for dissolution, uniformly mixing, adding phosphoric acid (1.4) for regulating the pH value to 3.1, and carrying out suction filtration through a 0.45 mu m filter membrane.
1.10 standard stock solutions azelaic acid and potassium azelate standard stock solutions (ρ=1 g/L): 0.01g (accurate to 0.0001 g) of azelaic acid and potassium azelate standard substance (1.1) are respectively weighed precisely in a 10mL measuring flask, dissolved by methanol (1.3) and fixed to volume to scale, and shaken uniformly to be used as a standard stock solution of azelaic acid and potassium azelate standard stock solution. Placing in a refrigerator at-18deg.C, and storing in dark place.
1.11 standard series solution the standard stock solution (1.10) 0.10mL, 0.50mL, 1.00mL, 2.00mL, 5.00mL and to 10mL measuring flask were accurately removed, the volume was fixed to the scale with methanol (1.3), and the mixture was shaken well to prepare standard series solutions with concentrations of 10.0. Mu.g/mL, 50.0. Mu.g/mL, 100.0. Mu.g/mL, 200.0. Mu.g/mL and 500. Mu.g/mL.
2. Apparatus and device
2.1 high performance liquid chromatograph, ultraviolet detector.
2.2 analytical balance: the sensing amount is 0.0001g and 0.00001 g.
2.3 high-speed centrifuge (rotational speed not less than 10000 rpm).
2.4 vortex mixer.
2.5 ultrasonic cleaner
3. Sample processing
0.20g (accurate to 0.001 g) of the sample is weighed and placed in a 50mL colorimetric tube with a plug, 45mL of sodium hydroxide aqueous solution (60 mmol/L) -methanol solution (1+1) (1.8) is added, vortex treatment is carried out for 1min, then the volume of the sodium hydroxide aqueous solution (60 mmol/L) -methanol solution (1+1) (1.8) is added to 50mL, ultrasonic treatment is carried out for 90min at 60 ℃, and centrifugation is carried out for 5min at 10000rpm, so as to obtain a solution to be tested.
4. Detection of
Sample solution and standard series solution containing azelaic acid and potassium azelate are connected to a high performance liquid chromatography system for detection.
High performance liquid chromatography conditions:
chromatographic column: b362V1 swedish SVEAC8 column (250 mm×4.6mm×5 μm), or equivalent chromatography column;
column temperature: 30 ℃;
mobile phase: solution a: phosphate buffer solution (1.9); solution B: acetonitrile (1.2), gradient elution procedure is shown in table 1.
TABLE 1 gradient elution procedure
Flow rate: 1.00mL/min;
sample injection amount: 10. Mu.L;
detection wavelength: 210nm.
Example 1
Labeling and measuring positive samples: three positive samples containing azelaic acid or potassium azelate are selected, positive standard adding test is carried out on the positive samples, components to be detected in an amount equivalent to the samples are added, and recovery rate results are measured. The sample information is detailed in Table 2.
TABLE 2 sample information
In example 1, the main instruments, materials and reagents used included:
1. instrument and reagent
Agilent1260 high performance liquid chromatograph (Agilent company, usa); sartorius CP224S and 225D-1CN electronic balances (certolis corporation, germany); 5800 ultrasonic apparatus (Branson Corp., U.S.); MS3 vortex mixer (IKA company, germany); 5810R-desk centrifuge (Eppendorf, germany); milli-Q Reference A+ type ultra-pure water instrument (Millipore Co., U.S.A.).
Azelaic acid standard, purity 99% (mass fraction, m/m, aledine); potassium nonyldiglycolate standard, purity 58.812% (mass fraction, m/m, shanghai microphone Biotechnology Co., ltd.) acetonitrile, methanol (chromatographic purity, merk, germany); phosphoric acid, sodium hydroxide, potassium dihydrogen phosphate (analytically pure, shanghai Lingfeng chemical Co., ltd.). The water is ultrapure water.
2. Chromatographic conditions
Chromatographic column: b362V1 swedish SVEAC8 column (250 mm×4.6mm×5 μm), or equivalent chromatography column;
column temperature: 30 ℃;
mobile phase: solution a: phosphate buffer solution; solution B: acetonitrile, gradient elution procedure is shown in table 1.
TABLE 1 gradient elution procedure
Phosphate buffer solution: accurately weighing 2.0401g of potassium dihydrogen phosphate, adding 1000mL of ultrapure water for dissolution, mixing, adding phosphoric acid (1.4) for regulating pH to 3.1, and filtering with a 0.45 μm filter membrane.
Flow rate: 1.00mL/min;
sample injection amount: 10. Mu.L;
detection wavelength: 210nm.
Linear relationship determination in hplc system:
standard stock solutions of azelaic acid and potassium azelate (ρ=1 g/L): 0.01g (accurate to 0.0001 g) of azelaic acid and potassium azelate standard substance are respectively weighed precisely in a 10mL measuring flask, dissolved by methanol, fixed to volume to scale, and shaken uniformly to be used as azelaic acid and potassium azelate standard stock solution. Placing in a refrigerator at-18deg.C, and storing in dark place.
The standard series solution is accurately transferred into measuring bottles of 0.10mL, 0.50mL, 1.00mL, 2.00mL, 5.00mL and 10mL of standard stock solution, methanol is used for metering to a scale, and the standard series solution with the concentration of 10.0 mug/mL, 50.0 mug/mL, 100.0 mug/mL, 200.0 mug/mL and 500 mug/mL is prepared by shaking evenly.
Linear regression analysis was performed with peak area on the ordinate (y) and concentration on the abscissa (x, μg/mL) to give the linear equation as shown in table 3 below.
TABLE 3 standard curve for azelaic acid and potassium nonyldiglycolate
4. Sample solution treatment of positive sample adding standard recovery rate: respectively taking 0.1g (accurate to 0.001 g) of azelaic acid positive samples and azelaic acid potassium-bisglycinate positive samples, respectively precisely adding azelaic acid or azelaic acid potassium-bisglycinate standard products which are equivalent to the samples in quantity, placing the samples in a 50mL colorimetric tube with a plug, adding 45mL of sodium hydroxide aqueous solution (60 mmol/L) -methanol solution (1+1)), carrying out vortex treatment for 1min, adding sodium hydroxide aqueous solution (60 mmol/L) -methanol solution (1+1) to fix the volume to 50mL, carrying out ultrasonic treatment at 60 ℃ for 90min, and centrifuging at 10000rpm for 5min to obtain a solution to be detected. Test solutions were prepared as recovery solutions by the method, and 6 experiments were repeated for each standard level, and recovery and relative standard deviation (n=6) were calculated.
Aqueous sodium hydroxide (60 mmol/L): weighing 2.4g of sodium hydroxide, adding 1000mL of water, dissolving and uniformly mixing.
Aqueous sodium hydroxide (60 mmol/L) -methanol solution (1+1): 100mL of an aqueous sodium hydroxide solution (60 mmol/L) was taken, 100mL of methanol (1.3) was added thereto, and the mixture was homogenized.
The calculation of azelaic acid and potassium azelate in the sample was calculated according to formula (1):
(1)
wherein: omega-the content of azelaic acid and potassium azelate in cosmetics,%;
m-sample amount, g;
ρ -the mass concentration of azelaic acid and potassium azelate in the cosmetics in the sample calculated by substituting the standard curve, μg/mL;
v-constant volume, mL.
Under the chromatographic condition, the standard working curve solution and the sample solution are taken and respectively injected, and the quality is improved according to the retention time and the spectrogram of the diode array. The contents of azelaic acid and potassium azelate in the sample solution were determined from the standard curve. The response values of azelaic acid and potassium azelate in the sample solution are in the linear range of the standard curve, and when the response values are beyond the linear range, the extracting solution is diluted and then the amount of the extracting solution is measured or increased for re-detection.
The results of the positive addition of the standard recovery measurements are shown in Table 4, which shows that the recovery measurements gave satisfactory results.
TABLE 4 recovery of positive samples and results of precision test
Example 2
Labeling and measuring a blank sample: taking three blank matrix samples of gel, cream emulsion and water which do not contain azelaic acid and potassium azelate and performing blank marking test on the three blank matrix samples, and determining recovery rate results.
In example 2, the main instruments, materials and reagents used included:
1. instrument and reagent
As in example 1 above.
2. Chromatographic conditions
As in example 1 above.
Linear relationship determination in HPLC System
Standard stock solutions of azelaic acid and potassium azelate (ρ=1 g/L): 0.01g (accurate to 0.0001 g) of azelaic acid and potassium azelate standard substance are respectively weighed precisely in a 10mL measuring flask, dissolved by methanol, fixed to volume to scale, and shaken uniformly to be used as azelaic acid and potassium azelate standard stock solution. Placing in a refrigerator at-18deg.C, and storing in dark place.
Accurately transferring the standard stock solution (1.10) into measuring bottles of 0.10mL, 0.50mL, 1.00mL, 2.00mL, 5.00mL and 10mL, fixing the volume to the scale by using methanol (1.3), and shaking uniformly to prepare standard series solutions with the concentration of 10.0 mug/mL, 50.0 mug/mL, 100.0 mug/mL, 200.0 mug/mL and 500 mug/mL.
Linear regression analysis was performed with peak area on the ordinate (y) and concentration on the abscissa (x, μg/mL) to give the linear equation as shown in table 5 below.
TABLE 5 standard curve for azelaic acid and potassium nonyldiglycolate
4. Blank sample marking recovery rate sample treatment: the method comprises the steps of respectively weighing 0.20g (accurate to 0.001 g) of a blank matrix sample of gel, cream emulsion and water, placing the blank matrix sample in a 50mL colorimetric tube with a plug, precisely adding a certain amount of azelaic acid and a certain amount of potassium azelaic acid standard substance, adding 45mL of sodium hydroxide aqueous solution (60 mmol/L) -methanol solution (1+1)), carrying out vortex treatment for 1min, adding the sodium hydroxide aqueous solution (60 mmol/L) -methanol solution (1+1) to fix the volume to 50mL, carrying out ultrasonic treatment at 60 ℃ for 90min, and centrifuging at 10000rpm for 5min to obtain a solution to be measured. Each labeling level was repeated 6 times to calculate recovery and relative standard deviation (n=6).
Aqueous sodium hydroxide (60 mmol/L): weighing 2.4g of sodium hydroxide, adding 1000mL of water, dissolving and uniformly mixing.
Aqueous sodium hydroxide (60 mmol/L) -methanol solution (1+1): 100mL of an aqueous sodium hydroxide solution (60 mmol/L) was taken, 100mL of methanol (1.3) was added thereto, and the mixture was homogenized.
The calculation of azelaic acid and potassium azelate in the sample was calculated according to formula (1):
(1)
wherein: omega-the content of azelaic acid and potassium azelate in cosmetics,%;
m-sample amount, g;
ρ -the mass concentration of azelaic acid and potassium azelate in the cosmetics in the sample calculated by substituting the standard curve, μg/mL;
v-constant volume, mL.
Under the chromatographic condition, the standard working curve solution and the sample solution are taken and respectively injected, and the quality is improved according to the retention time and the spectrogram of the diode array. The contents of azelaic acid and potassium azelate in the sample solution were determined from the standard curve. The response values of azelaic acid and potassium azelate in the sample solution are in the linear range of the standard curve, and when the response values are beyond the linear range, the extracting solution is diluted and then the amount of the extracting solution is measured or increased for re-detection.
The results of the blank matrix labeling recovery measurements are shown in Table 6, which shows that the recovery measurements gave satisfactory results.
TABLE 6 recovery of blank matrix addition and results of precision test
Example 3
Cosmetic 1 containing azelaic acid and cosmetic 2 containing potassium azelate in factory A and factory B are extracted and detected respectively: the azelaic acid mark content in the A factory sample is 20%; 5% of potassium azetidyl diglycine in the B plant sample.
In example 3, the main instruments, materials and reagents used included:
1. instrument and reagent
As in example 1 above.
2. Chromatographic conditions
As in example 1 above.
Linear relationship determination in hplc system:
standard stock solutions of azelaic acid and potassium azelate (ρ=1 g/L): 0.01g (accurate to 0.0001 g) of azelaic acid and potassium azelate standard substance are respectively weighed precisely in a 10mL measuring flask, dissolved by methanol, fixed to volume to scale, and shaken uniformly to be used as azelaic acid and potassium azelate standard stock solution. Placing in a refrigerator at-18deg.C, and storing in dark place.
Accurately transferring the standard stock solution (1.10) into measuring bottles of 0.10mL, 0.50mL, 1.00mL, 2.00mL, 5.00mL and 10mL, fixing the volume to the scale by using methanol (1.3), and shaking uniformly to prepare standard series solutions with the concentration of 10.0 mug/mL, 50.0 mug/mL, 100.0 mug/mL, 200.0 mug/mL and 500 mug/mL.
Linear regression analysis was performed with peak area on the ordinate (y) and concentration on the abscissa (x, μg/mL) to give the linear equation as shown in table 7 below.
TABLE 7 Linear relationship table of azelaic acid and potassium azelate
4. Sample processing
0.20g (accurate to 0.001 g) of the sample is weighed and placed in a 50mL colorimetric tube with a plug, 45mL of sodium hydroxide aqueous solution (60 mmol/L) -methanol solution (1+1)) is added, vortex treatment is carried out for 1min, then sodium hydroxide aqueous solution (60 mmol/L) -methanol solution (1+1) is added to fix the volume to 50mL, after ultrasonic treatment for 90min at 60 ℃,10000rpm centrifugation is carried out for 5min, and the sample is taken as the solution to be measured.
Aqueous sodium hydroxide (60 mmol/L): weighing 2.4g of sodium hydroxide, adding 1000mL of water, dissolving and uniformly mixing.
Aqueous sodium hydroxide (60 mmol/L) -methanol solution (1+1): 100mL of an aqueous sodium hydroxide solution (60 mmol/L) was taken, 100mL of methanol was added thereto, and the mixture was homogenized.
Under the chromatographic condition, the standard working curve solution and the sample solution are taken and respectively injected, and the quality is improved according to the retention time and the spectrogram of the diode array. The contents of azelaic acid and potassium azelate in the sample solution were determined from the standard curve. The response values of azelaic acid and potassium azelate in the sample solution are in the linear range of the standard curve, and when the response values are beyond the linear range, the extracting solution is diluted and then the amount of the extracting solution is measured or increased for re-detection.
The calculation of azelaic acid and potassium azelate in the sample was calculated according to formula (1):
(1)
wherein: omega-the content of azelaic acid and potassium azelate in cosmetics,%;
m-sample amount, g;
ρ -the mass concentration of azelaic acid and potassium azelate in the cosmetics in the sample calculated by substituting the standard curve, μg/mL;
v-constant volume, mL.
The sample measurement results are shown in Table 8, and the extraction method and the rapid measurement method can accurately quantify the contents of azelaic acid and potassium azelaic acid in the sample.
TABLE 8 determination of azelaic acid in samples
In conclusion, the detection method of azelaic acid and potassium azelate in the cosmetics is simple and quick to operate, high in sensitivity, and the recovery rate and the repeatability meet the requirements of daily detection. The method has high sensitivity, high quality accuracy and feasible linear range, and greatly improves the monitoring capability of the detection mechanism on cosmetics.
The foregoing is merely illustrative of the preferred embodiments of the present invention and is not intended to limit the embodiments and scope of the present invention, and it should be appreciated by those skilled in the art that equivalent substitutions and obvious variations may be made using the teachings and illustrations of the present invention, and that such variations are intended to be included within the scope of the present invention.
Claims (7)
1. A method for detecting azelaic acid and potassium azelate in cosmetics, which is characterized by comprising the following steps:
step one, preparing standard series solutions: precisely weighing azelaic acid and potassium azelate standard substances in a measuring flask respectively, dissolving the azelaic acid and the potassium azelate standard substances in methanol, fixing the volume to a scale, shaking the solution uniformly to serve as an azelaic acid and potassium azelate standard stock solution, accurately transferring the standard stock solution, fixing the volume by methanol, and preparing to obtain a standard series of solutions;
step two, preparing a solution to be measured: precisely weighing a sample to be measured, adding a sodium hydroxide aqueous solution-methanol solution, performing vortex treatment, adding the sodium hydroxide aqueous solution-methanol solution to fix the volume, performing ultrasonic treatment at 50-60 ℃ for 60-90min, and centrifuging for 5-20min to obtain a solution to be measured;
and thirdly, connecting the standard series solution prepared in the first step and the solution to be detected prepared in the second step to a high performance liquid chromatography system for detection.
2. The method according to claim 1, wherein the standard series of solutions have respective concentrations of: 10.0. Mu.g/mL, 50.0. Mu.g/mL, 100.0. Mu.g/mL, 200.0. Mu.g/mL, 500. Mu.g/mL.
3. The detection method according to claim 1, wherein the specific preparation method of the solution to be detected is as follows: weighing 0.20g of a sample to be measured, placing the sample into a 50mL colorimetric tube with a plug, adding 45mL of sodium hydroxide aqueous solution-methanol solution, performing vortex treatment for 1min, adding the sodium hydroxide aqueous solution-methanol solution to fix the volume to 50mL, performing ultrasonic treatment at 60 ℃ for 90min, and centrifuging at 10000rpm for 5min to obtain the solution to be measured.
4. The method according to claim 1, wherein the conditions of high performance liquid chromatography:
chromatographic column: B362V1 Swedish SVEAC8 column, 250mm×4.6mm×5 μm;
column temperature: 30 ℃;
mobile phase: solution A is phosphate buffer solution; solution B is acetonitrile; gradient elution;
flow rate: 1.00mL/min;
sample injection amount: 10. Mu.L;
detection wavelength: 210nm.
5. The method according to claim 4, wherein the gradient elution procedure is as follows:
6. the method according to claim 4, wherein the phosphate buffer solution is prepared by the following steps: precisely weighing 2.0-2.5g of potassium dihydrogen phosphate, adding 1000mL of ultrapure water for dissolution, mixing uniformly, adding phosphoric acid for regulating the pH to 3.1, and carrying out suction filtration through a 0.45 mu m filter membrane.
7. The method according to claim 6, wherein the mass concentration of the phosphoric acid is 85%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210682555.3A CN115219613B (en) | 2022-06-16 | 2022-06-16 | Method for detecting azelaic acid and potassium azelate in cosmetics |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210682555.3A CN115219613B (en) | 2022-06-16 | 2022-06-16 | Method for detecting azelaic acid and potassium azelate in cosmetics |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115219613A CN115219613A (en) | 2022-10-21 |
| CN115219613B true CN115219613B (en) | 2023-12-22 |
Family
ID=83607658
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210682555.3A Active CN115219613B (en) | 2022-06-16 | 2022-06-16 | Method for detecting azelaic acid and potassium azelate in cosmetics |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115219613B (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010117258A1 (en) * | 2009-04-09 | 2010-10-14 | Malaysian Palm Oil Board | A method of synthesising an amino acid derivative of azelaic acid |
| CN102018766A (en) * | 2009-09-16 | 2011-04-20 | 博仲盛景医药技术（北京）有限公司 | Plant extractive, extraction method and application thereof as well as composite comprising same |
| CN103822976A (en) * | 2013-10-15 | 2014-05-28 | 辽宁省食品药品检验所 | Method for measuring 4-potassium methoxysalicylate in cosmetics |
| CN106033077A (en) * | 2015-03-12 | 2016-10-19 | 谱尼测试集团上海有限公司 | Detection method for determining kojic acid content in cosmetics |
| CN109655552A (en) * | 2019-01-28 | 2019-04-19 | 深圳市药品检验研究院（深圳市医疗器械检测中心） | A method of with o-cymene -5- alcohol in Gas Chromatography-Mass Spectrometry cosmetics |
| CN110464683A (en) * | 2018-05-09 | 2019-11-19 | 江阴云智医疗无纺布制品有限公司 | A kind of wet tissue liquid formulations and preparation method thereof that effective control small pox generates |
| CN112684050A (en) * | 2020-12-24 | 2021-04-20 | 上海微谱检测技术有限公司 | Research on determination method of whitening functional components in cosmetics |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20180021329A1 (en) * | 2015-02-13 | 2018-01-25 | Maruho Co., Ltd. | External preparation comprising pyridonecarboxylic acid derivative |
| CN109298115B (en) * | 2018-10-19 | 2020-07-28 | 深圳市绘云生物科技有限公司 | Quantitative detection method for multiple metabolites in biological sample and metabolic chip |
-
2022
- 2022-06-16 CN CN202210682555.3A patent/CN115219613B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010117258A1 (en) * | 2009-04-09 | 2010-10-14 | Malaysian Palm Oil Board | A method of synthesising an amino acid derivative of azelaic acid |
| CN102018766A (en) * | 2009-09-16 | 2011-04-20 | 博仲盛景医药技术（北京）有限公司 | Plant extractive, extraction method and application thereof as well as composite comprising same |
| CN103822976A (en) * | 2013-10-15 | 2014-05-28 | 辽宁省食品药品检验所 | Method for measuring 4-potassium methoxysalicylate in cosmetics |
| CN106033077A (en) * | 2015-03-12 | 2016-10-19 | 谱尼测试集团上海有限公司 | Detection method for determining kojic acid content in cosmetics |
| CN110464683A (en) * | 2018-05-09 | 2019-11-19 | 江阴云智医疗无纺布制品有限公司 | A kind of wet tissue liquid formulations and preparation method thereof that effective control small pox generates |
| CN109655552A (en) * | 2019-01-28 | 2019-04-19 | 深圳市药品检验研究院（深圳市医疗器械检测中心） | A method of with o-cymene -5- alcohol in Gas Chromatography-Mass Spectrometry cosmetics |
| CN112684050A (en) * | 2020-12-24 | 2021-04-20 | 上海微谱检测技术有限公司 | Research on determination method of whitening functional components in cosmetics |
Non-Patent Citations (10)
| Title |
|---|
| Analysis of aliphatic dicarboxylic acids in pharmaceuticals and cosmetics by liquid chromatography （ HPLC ） with fluorescence detection;GATTI R;《Pharm Biomed Anal》;第13卷(第45期);全文 * |
| On column approach in the HPLC UV analysis of non chromophoric compounds using azelaic acid as a model;ALZWEIRI M;《Jordan J Pharm Sci》;第5卷(第3期);全文 * |
| Spectrophotometric determination of azelaic acid in pharmaceutical formulations;KISHORE M;《J Pharm Res》;第3卷(第12期);全文 * |
| Validated stability indicating RP HPLC method for analysis of azelaic acid in pharmaceuticals;MALIK D S;《Indian J Pharm Sci》;第80卷(第3期);全文 * |
| 化妆品中壬二酸等6种化合物的测定;韩晶;《中国卫生检验杂志》;第32卷(第19期);全文 * |
| 壬二酸凝胶的制备与质量控制;刘丹;《中国医药科学》;第10卷(第19期);全文 * |
| 气相色谱法测定化妆品中的壬二酸;周耀斌;赵艳菊;周静;杨丹;杨保刚;;香料香精化妆品(第06期);全文 * |
| 高效液相色谱法同时测定化妆品中12种美白成分;简龙海;陈丹丹;茹歌;许勇;勇郑;荣王柯;;香料香精化妆品(第06期);全文 * |
| 高效液相色谱法同时测定化妆品中壬二酸及其衍生物壬二酰二甘氨酸钾的含量;许勇;《上海预防医学》;第34卷(第12期);全文 * |
| 高效液相色谱法同时测定复方壬二酸乳膏中维生素C与维生素E的含量;臧亚茹;;中国医院药学杂志(第08期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115219613A (en) | 2022-10-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104597160B (en) | HPLC (High Performance Liquid Chromatography) method for simultaneously determining content of six organic acids in pinellia ternata | |
| CN107202836B (en) | Method for rapidly analyzing theanine content in fresh tea sample | |
| CN106525997A (en) | Method for determination of organic acids and flavone components in polygonum viviparum | |
| CN102955010A (en) | Quality detection method of traditional Chinese medicine callicarpa nudiflora preparation | |
| CN106645518B (en) | The measuring method of chloramphenicol residue in a kind of propolis virgin rubber | |
| CN110146604A (en) | The analysis method of low content sodium pyrosulfite in a kind of measurement sustained release pharmaceutical formulation | |
| CN109856270A (en) | A method of with 7 index components in hplc simultaneous determination canopy powder granule | |
| CN103630618A (en) | Method for determining betaine content in wolfberry | |
| CN115219613B (en) | Method for detecting azelaic acid and potassium azelate in cosmetics | |
| CN102706971A (en) | A method for analysis of corticosteroids in cosmetics utilizing reverse microemulsion electrokinetic chromatography by taking an ionic liquid as an additive | |
| CN106706835A (en) | Quality detection method of trollius chinensis bunge effervescent tablets | |
| CN102967684A (en) | Yanning capsule quality standard detection method | |
| CN102078503A (en) | Detection method for pulse-activating decoction traditional Chinese medicine preparation | |
| CN111289655B (en) | Analysis method for simultaneously determining PCA and t-UCA in skin stratum corneum patch | |
| CN103901147A (en) | Method for measuring dissolution rate of andrographolide pills | |
| CN108037200B (en) | Quality detection method of kidney nourishing and tranquilizing pills | |
| CN107807182B (en) | Method for measuring content of ganoderic acid A in ganoderma lucidum syrup | |
| CN103175930A (en) | High performance liquid chromatography analysis method for measuring sodium sulfite content | |
| CN112946129A (en) | Quality detection method of antidiarrheal syrup | |
| CN109085273A (en) | The high performance liquid chromatography while method for separating and detecting of 12 kinds of direct dyess of cosmetics | |
| CN104931325B (en) | The extracting method of incretion interferent | |
| CN109100457A (en) | The detection method of 1-Deoxynojirimycin content in a kind of mulberry-leaf extract | |
| CN115236222B (en) | Method for detecting human epidermal growth factor in cosmetics | |
| CN102944618A (en) | Content detection method of Chinese medicine preparation for treating gastropathy | |
| CN102854283A (en) | Detection method of polygala arvensis |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |