CN115792025A - Method for detecting acid red 92 in cosmetics - Google Patents
Method for detecting acid red 92 in cosmetics Download PDFInfo
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- CN115792025A CN115792025A CN202211595135.8A CN202211595135A CN115792025A CN 115792025 A CN115792025 A CN 115792025A CN 202211595135 A CN202211595135 A CN 202211595135A CN 115792025 A CN115792025 A CN 115792025A
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- methanol
- ammonium acetate
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- acid red
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- OOYIOIOOWUGAHD-UHFFFAOYSA-L disodium;2',4',5',7'-tetrabromo-4,5,6,7-tetrachloro-3-oxospiro[2-benzofuran-1,9'-xanthene]-3',6'-diolate Chemical compound [Na+].[Na+].O1C(=O)C(C(=C(Cl)C(Cl)=C2Cl)Cl)=C2C21C1=CC(Br)=C([O-])C(Br)=C1OC1=C(Br)C([O-])=C(Br)C=C21 OOYIOIOOWUGAHD-UHFFFAOYSA-L 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 45
- 239000002537 cosmetic Substances 0.000 title claims abstract description 28
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 132
- 239000000126 substance Substances 0.000 claims abstract description 26
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 19
- 239000012085 test solution Substances 0.000 claims abstract description 11
- 238000001914 filtration Methods 0.000 claims abstract description 9
- 238000005303 weighing Methods 0.000 claims abstract description 8
- 238000000605 extraction Methods 0.000 claims abstract description 7
- 239000011259 mixed solution Substances 0.000 claims abstract description 7
- 239000006228 supernatant Substances 0.000 claims abstract description 7
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 28
- 239000005695 Ammonium acetate Substances 0.000 claims description 28
- 229940043376 ammonium acetate Drugs 0.000 claims description 28
- 235000019257 ammonium acetate Nutrition 0.000 claims description 28
- 238000001514 detection method Methods 0.000 claims description 13
- 239000000523 sample Substances 0.000 claims description 10
- 239000012488 sample solution Substances 0.000 claims description 9
- 238000010828 elution Methods 0.000 claims description 6
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 claims description 5
- 239000003643 water by type Substances 0.000 claims description 5
- 238000012790 confirmation Methods 0.000 claims description 3
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 3
- 239000010931 gold Substances 0.000 claims description 3
- 229910052737 gold Inorganic materials 0.000 claims description 3
- QEVHRUUCFGRFIF-MDEJGZGSSA-N reserpine Chemical compound O([C@H]1[C@@H]([C@H]([C@H]2C[C@@H]3C4=C(C5=CC=C(OC)C=C5N4)CCN3C[C@H]2C1)C(=O)OC)OC)C(=O)C1=CC(OC)=C(OC)C(OC)=C1 QEVHRUUCFGRFIF-MDEJGZGSSA-N 0.000 claims description 3
- 238000004587 chromatography analysis Methods 0.000 claims 2
- 238000010812 external standard method Methods 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 239000000047 product Substances 0.000 description 18
- 239000000243 solution Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 4
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 3
- 239000013076 target substance Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 239000002253 acid Substances 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000003973 paint Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- GJCOSYZMQJWQCA-UHFFFAOYSA-N 9H-xanthene Chemical compound C1=CC=C2CC3=CC=CC=C3OC2=C1 GJCOSYZMQJWQCA-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 235000015895 biscuits Nutrition 0.000 description 1
- 235000012970 cakes Nutrition 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000000976 ink Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- QDLAGTHXVHQKRE-UHFFFAOYSA-N lichenxanthone Natural products COC1=CC(O)=C2C(=O)C3=C(C)C=C(OC)C=C3OC2=C1 QDLAGTHXVHQKRE-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- 230000037307 sensitive skin Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
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Abstract
The application provides a method for detecting acid red 92 in cosmetics, which comprises the following steps: weighing the substance to be detected, and adding a first volume of methanol into each gram of the substance to be detected for extraction; centrifuging the mixed solution of the substance to be detected and the methanol, taking supernatant and filtering to obtain a sample test solution; injecting the sample test solution into a high performance liquid chromatography column, measuring at 485 by an ultraviolet detector, and calculating the content of acid red 92 by an external standard method, so that the use of the substances is standardized, and the risk is avoided for social production.
Description
Technical Field
The application relates to the technical field of analytical chemistry, in particular to a method for detecting acid red 92 in cosmetics.
Background
Acid Red 92 (Red 27/28) CL45410 is an artificial xanthene colorant, which is applied to candies, cakes, fish cakes, sausages, confectionery products and biscuits by some enterprises, can also be used for coloring coatings, paint printing, paints, inks, plastic products, cultural and educational products and the like, is widely applied to the fields of food and cosmetics, and has high safety factor, but excessive acid Red 92 can induce cell necrosis and initiate inflammatory reaction, and is used with caution by consumers with pox-causing property and sensitive skin.
The prior art does not relate to a detection method of acid red 92, and a detection method for detecting the content of acid red 92 is needed.
Disclosure of Invention
In view of the problem, the present application is proposed in order to provide a method for detecting acid red 92 in a cosmetic product that overcomes or at least partially solves the problem, comprising:
a method for detecting acid red 92 in cosmetics comprises the following steps:
weighing the substance to be detected, and adding a first volume of methanol into each gram of the substance to be detected for extraction;
centrifuging the mixed solution of the substance to be detected and the methanol, taking supernatant and filtering to obtain a sample test solution;
and injecting the sample test solution into a high performance liquid chromatography column, and measuring at 485 by an ultraviolet detector.
Further, the first volume is 10-15mL.
Further, the filtration was performed with a purple filter.
Further, the column model was Thermo Hypersil GOLD C18.
Further, the step of measuring the upper computer test solution by high performance liquid chromatography is carried out, and mobile phases are 20mmol/L ammonium acetate and methanol.
Further, the elution gradient was: at 0min, the ammonium acetate is 90 percent, and the methanol is 10 percent; at 14min, the ammonium acetate is 5% and the methanol is 95%; at 18min, the ammonium acetate is 5 percent, and the methanol is 95 percent; at 18.5min, the ammonium acetate is 90%, and the methanol is 10%; at 22min, the ammonium acetate is 90% and the methanol is 10%.
Further, the step of measuring the upper computer test solution by high performance liquid chromatography further comprises:
confirmation of the results was performed by LC/MS.
Further, the liquid chromatograph-mass spectrometer uses a chromatographic column with the model number of Waters BEH C18
Further, the step of confirmation by LC-MS was carried out with 20mmol/L ammonium acetate and methanol as mobile phases.
Further, the elution gradient was: at 0min, 70% of ammonium acetate and 30% of methanol; at 0.2min, the ammonium acetate is 70 percent, and the methanol is 30 percent; at 2min, the ammonium acetate is 0 percent, and the methanol is 100 percent; at 4min, the ammonium acetate is 0 percent, and the methanol is 100 percent; at 4.1min, 70% of ammonium acetate and 30% of methanol; at 6.5min, the ammonium acetate is 70% and the methanol is 30%.
The application has the following advantages:
in the embodiment of the application, the blank that the acid red detection is not available in the prior art is filled, and the application provides a method for detecting the acid red 92 in the cosmetics, which comprises the following steps: weighing the substance to be detected, and adding a first volume of methanol into each gram of the substance to be detected for extraction; centrifuging the mixed solution of the substance to be detected and the methanol, taking supernatant and filtering to obtain a sample test solution; injecting the sample test solution into a high performance liquid chromatography column, measuring at 485 by an ultraviolet detector, and calculating the content of acid red 92 by an external standard method, so that the use of the substances is standardized, and the risk is avoided for social production.
Drawings
In order to more clearly illustrate the technical solutions of the present application, the drawings needed to be used in the description of the present application will be briefly introduced below, and it is apparent that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without inventive labor.
FIG. 1 is a flowchart illustrating steps of a method for detecting acid Red 92 in a cosmetic product according to one embodiment of the present disclosure;
FIG. 2 is a flowchart illustrating the operation of a method for detecting acid Red 92 in a cosmetic product according to one embodiment of the present disclosure;
FIG. 3 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 0.5mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present application;
FIG. 4 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 1mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present disclosure;
FIG. 5 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 2mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present disclosure;
FIG. 6 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 5mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present disclosure;
FIG. 7 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 10mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present disclosure;
FIG. 8 is a line graph of acid Red 92 in a method for detecting acid Red 92 in a cosmetic product according to one embodiment of the present application;
FIG. 9 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 0.01mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present application;
FIG. 10 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 0.05mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present application;
FIG. 11 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 0.1mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present application;
FIG. 12 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 0.2mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present application;
FIG. 13 is a chromatogram of a standard curve for acid Red 92 at a concentration point of 0.5mg/L for a method of detecting acid Red 92 in a cosmetic product according to an embodiment of the present application;
FIG. 14 is a line graph of acid Red 92 in a method for detecting acid Red 92 in a cosmetic product according to an embodiment of the present application.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present application more comprehensible, the present application is described in further detail with reference to the accompanying drawings and the detailed description. It is to be understood that the embodiments described are only a few embodiments of the present application and not all embodiments. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments in the present application without making any creative effort belong to the protection scope of the present application.
The inventor discovers by analyzing the prior art that: after the object to be tested is subjected to certain pretreatment, the content of acid red 92 in the cosmetics can be measured by using high performance liquid chromatography through researching instrument parameters and improving instrument methods.
Referring to fig. 1-2, a flowchart illustrating steps and operational flow of a method for detecting acid red 92 in a cosmetic according to an embodiment of the present disclosure is shown:
the method comprises the following steps:
s110, weighing the substance to be detected, and adding methanol with a first volume per gram of the substance to be detected for extraction;
s120, centrifuging the mixed solution of the substance to be detected and the methanol, taking supernatant, and filtering to obtain a sample solution;
s130, injecting the sample solution into a high performance liquid chromatography column, and measuring at 485 through an ultraviolet detector.
In the embodiment of the application, the blank that the acid red detection is not available in the prior art is filled, and the application provides a method for detecting the acid red 92 in the cosmetics, which comprises the following steps: weighing the substance to be measured, and adding methanol with a first volume into each gram of the substance to be measured; centrifuging the mixed solution of the solution to be detected and the methanol, taking supernatant and filtering to obtain a sample test solution; injecting the sample test solution into a high performance liquid chromatography column, measuring at 485 by an ultraviolet detector, and calculating the content of acid red 92 by an external standard method, so that the use of the substances is standardized, and the risk is avoided for social production.
Next, a method for detecting acid red 92 in one cosmetic in the present exemplary embodiment will be further described.
Weighing the substance to be detected, and adding a first volume of methanol per gram of the substance to be detected for extraction as described in the step S110.
In an embodiment of the present invention, the "weighing the substance to be tested" in step S110 can be further described with reference to the following description, wherein a first volume of methanol is added to each gram of the substance to be tested; "is used herein.
It should be noted that the analyte is a solid fragment with a total amount of not more than 5 g. The object to be tested is a substance for extracting a small amount of target substance from the target object to be tested for detection, and the substance to be tested has one of effective ways for obtaining various data by testing the target object on the premise of not influencing the main properties of the target object, the extraction amount of the object to be tested is enough to carry out the test for 3-5 times, the sampling area selection process of the object to be tested is required to have randomness during sampling, and subjective selection cannot be doped in the selection process;
if the subsequent measurement result value is larger, the solid-liquid ratio can be properly adjusted to ensure that the object to be measured is fully extracted.
In one embodiment, 0.5g of sample is weighed and extracted with 5mL of methanol.
And step S120, centrifuging the mixed solution of the solution to be detected and the methanol, taking supernatant, and filtering to obtain a sample solution. Need adopt purple filter membrane to filter, it receives the influence of filter membrane to place content.
The sample solution is injected into a high performance liquid chromatography column and measured by an ultraviolet detector at 485 as described in step S130.
In an embodiment of the present invention, the specific process of "performing determination on the sample solution by high performance liquid chromatography" in step S130 can be further described with reference to the following description.
The instrument parameters are as follows:
high performance liquid chromatography:
the instrument comprises the following steps: HPLC-DAD (high-Performance liquid chromatography-diode array Detector)
Chromatographic column Thermo Hypersil GOLD C18, 250 x 4.6mm,5um;
mobile phase A:20mmol/L ammonium acetate; b: methanol;
the flow rate gradient elution procedure is shown in table 1:
| time/ | A | B | |
| 0 | 90 | 10 | |
| 14 | 5 | 95 | |
| 18 | 5 | 95 | |
| 18.5 | 90 | 10 | |
| 22 | 90 | 10 |
TABLE 1
Detection wavelength: 483nm;
column temperature: 30 ℃;
sample introduction amount: 20 mu L of the solution;
operating time: and (5) 22min.
Referring to fig. 3-8, the chromatograms of the various concentration points of the standard curve for acid red 92 and the linearity of acid red 92 determined by hplc, the linear range of the standard curve and the retention time of the target substance of acid red 92 are shown in table 2:
TABLE 2
The method comprises the following steps: confirming the result by a liquid chromatograph-mass spectrometer;
the instrument parameters are as follows:
the instrument comprises the following steps: waters TQD;
chromatographic column Waters BEH C18, 100X 2.1mm,1.7um;
mobile phase A:5mmol/L ammonium acetate; b: methanol;
the flow rate gradient elution procedure is shown in table 2:
| time/min | A(％) | B(％) |
| 0 | 70 | 30 |
| 0.2 | 70 | 30 |
| 2 | 0 | 100 |
| 4 | 0 | 100 |
| 4.1 | 70 | 30 |
| 6.5 | 70 | 30 |
TABLE 3
Sample introduction amount: 10 mu L of the solution;
column temperature: 30 ℃;
an ion source: ESI;
operating time: 5min;
the mass spectral information is shown in table 3:
TABLE 4
Referring to FIGS. 9-13, for the chromatograms of the various concentration points of the standard curve for acid Red 92 and the linearity of acid Red 92 determined using waters TQD, the linear range of the standard curve and the target substance retention time for acid Red 92 are shown in Table 5:
TABLE 5
FIG. 14 is a high performance liquid chromatography/high performance liquid chromatography tandem mass spectrometry data of a certain eye shadow, the spectrogram shows that the method can realize the characteristics of high-efficiency separation, stable baseline and the like; recovery rates were in the satisfactory range of 80% -110% by applying a standard recovery test to the eye shadow samples. Has good repeatability and reproducibility and accurate quantification.
According to the embodiment, the acid red 92 is detected by using HPLC (high performance liquid chromatography) and LC-MS/MS (liquid chromatography tandem mass spectrometry), and HPLC and LCMSMS are adopted, so that the method has the advantages of excellent repeatability and reproducibility, simplicity and convenience in operation, high speed, easiness in obtaining of used equipment and high popularization.
While preferred embodiments of the present application have been described, additional variations and modifications of these embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including the preferred embodiment and all changes and modifications that fall within the true scope of the embodiments of the present application.
Finally, it should also be noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or terminal that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or terminal. Without further limitation, an element defined by the phrase "comprising a … …" does not exclude the presence of another identical element in a process, method, article, or terminal device that comprises the element.
The method for detecting acid red 92 in cosmetics provided by the present application is described in detail above, and the principle and the embodiment of the present application are explained in the present application by applying specific examples, and the description of the above examples is only used to help understanding the method and the core idea of the present application; meanwhile, for a person skilled in the art, according to the idea of the present application, there may be variations in the specific embodiments and the application scope, and in summary, the content of the present specification should not be construed as a limitation to the present application.
Claims (10)
1. A method for detecting acid red 92 in a cosmetic, comprising the steps of:
weighing the substance to be detected, and adding a first volume of methanol into each gram of the substance to be detected for extraction;
centrifuging the mixed solution of the substance to be detected and the methanol, taking supernatant and filtering to obtain a sample solution;
and injecting the sample test solution into a high performance liquid chromatography column, and measuring at 485 by an ultraviolet detector.
2. The detection method according to claim 1, wherein the first volume is 10-15mL.
3. The detection method according to claim 1, wherein the filtration is performed with a purple filter.
4. The method of claim 1, wherein the chromatography column is Thermo Hypersil GOLD C18.
5. The detection method according to claim 1, wherein the step of injecting the sample solution into a high performance liquid chromatography column and measuring the sample solution at 485 by an ultraviolet detector has a mobile phase of 20mmol/L ammonium acetate and methanol.
6. The detection method according to claim 5, wherein the elution gradient is: at 0min, the ammonium acetate is 90 percent, and the methanol is 10 percent; at 14min, the ammonium acetate is 5% and the methanol is 95%; at 18min, the ammonium acetate is 5% and the methanol is 95%; at 18.5min, the ammonium acetate is 90%, and the methanol is 10%; at 22min, the ammonium acetate is 90% and the methanol is 10%.
7. The method of claim 1, wherein the step of measuring the upper sample solution by HPLC further comprises:
confirmation of the results was performed by LC MS.
8. The detection method according to claim 7, wherein the chromatography column model used by the LC-MS is Waters BEH C18.
9. The detection method according to claim 7, wherein the step of confirming is performed by a LC-MS, and the mobile phase is 20mmol/L ammonium acetate and methanol.
10. The detection method according to claim 9, wherein the elution gradient is: at 0min, the ammonium acetate is 70 percent, and the methanol is 30 percent; at 0.2min, the ammonium acetate is 70 percent, and the methanol is 30 percent; at 2min, the ammonium acetate is 0 percent, and the methanol is 100 percent; at 4min, the ammonium acetate is 0 percent, and the methanol is 100 percent; at 4.1min, the ammonium acetate is 70 percent, and the methanol is 30 percent; at 6.5min, the ammonium acetate is 70% and the methanol is 30%.
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Cited By (1)
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