Method for detecting hair dye components in hair dye product

Abstract

The invention discloses a method for detecting hair dye components in a hair dyeing product, which comprises the following steps: step 1, adding a supramolecular solvent into a sample to be detected, and performing dispersion liquid-liquid extraction to obtain an extracting solution to be detected; wherein the supramolecular solvent is prepared from alkyl alcohol, tetrahydrofuran and water; and step 2, determining the contents of the tetraaminopyrimidine sulfate and the 2, 6-dimethoxy-3, 5-pyridine diamine hydrochloride in the extracting solution to be detected by using a liquid chromatography-tandem mass spectrometry method. The pretreatment operation is simple and rapid, the cost is low, the enrichment efficiency is high, and the dosage of an organic solvent is small; the specificity is good, the sensitivity is higher, and the sample analysis time is shortened; the device has the advantages of integrating extraction and purification, being green and environment-friendly and the like.

Description

Technical Field

[0001] The invention relates to a method for detecting hair dye components in hair dye products. Background Art

[0002] As a cosmetic product that changes hair color, hair dye products can, on the one hand, make hair color return to its natural state, and middle-aged and elderly people with white or grey hair can dye their hair to make them look younger; on the other hand, they can be used as a personal and fashionable embellishment. In recent years, the global hair dye market has seen strong demand, and it has become a relatively active product in the cosmetics field, indicating that the hair dye market will have greater development in the future.

[0003] Some ingredients in hair dyes, especially oxidative hair dyes, have certain safety risks. Developed countries such as Europe, America, and Japan have used hair dyes for decades, and their potential dangers have been exposed. For example, aromatic amines and phenol compounds have varying degrees of sensitization, mutagenicity, and other potential toxicity. Some chemicals in hair dyes may cause local skin or systemic allergies. Inhalation of their dust may cause allergies, rhinitis, bronchitis, fever, and other different symptoms, and may also cause damage to the liver and urinary system. Long-term exposure to hair dyes may increase the incidence of cancer and is potentially dangerous.

[0004] Therefore, the health problems caused by the frequent use of hair dye products have always received widespread attention. The safety of dye ingredients has been increasingly valued by consumers and regulatory authorities. All countries have conducted risk assessments on hair dyes and made corresponding regulations and restrictions on their use. Table 7 of the "Technical Specifications for Safety of Cosmetics" (2015 edition) lists 75 dyes that are allowed in hair dye cosmetics, and clarifies their restricted use requirements and labeling regulations. For some raw materials, warnings are also required. The above regulations show that the country attaches great importance to the safe use of hair dyes. However, the quality of hair dye products on the market is uneven. Most of the unqualified cosmetics in the supervision and random inspection announcements issued by the State Food and Drug Administration in 2018 and 2019 are hair dye products, mainly involving hair dye content exceeding the prescribed limit, hair dyes not marked in the approval documents, hair dyes not marked on the labels, and labels inconsistent with approval documents. The current testing standards cannot cover all hair dyes, and there may be more unqualified products in reality. Hair dyes are still the focus of supervision. Although the hair dyes covered in the current testing standards already account for most of the "Technical Specifications for Safety of Cosmetics" (2015 edition), there are still gaps in some components including tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride, which cannot fully meet the needs of regulating hair dye products. We should continue to establish rapid and effective analysis methods and gradually improve the supporting testing system. Summary of the invention

[0005] The technical problem to be solved by the present invention is to overcome the above-mentioned defects in the prior art and provide a method for detecting hair dye ingredients in hair dye products.

[0006] The present invention solves the above technical problems through the following technical solutions:

[0007] A method for detecting hair dye ingredients in a hair dye product comprises the following steps:

[0008] Step 1, adding a supramolecular solvent to a sample to be detected, and performing dispersed liquid-liquid extraction to obtain an extract to be detected; wherein the supramolecular solvent is composed of an alkyl alcohol, tetrahydrofuran and water;

[0009] Step 2, determining the contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be tested by liquid chromatography-tandem mass spectrometry.

[0010] Step 3, preparing a series of standard solutions of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride with concentration gradients, detecting them by liquid chromatography-tandem mass spectrometry sample analysis method, and establishing a chromatographic peak area-concentration standard correspondence relationship according to the detection results;

[0011] Furthermore, in step 1, the alkyl alcohol in the supramolecular solvent is n-decanol.

[0012] Furthermore, the ratio of n-decanol, tetrahydrofuran and water in the supramolecular solvent is 1:5:40.

[0013] Furthermore, in step 1, the dispersed liquid-liquid extraction step includes: vortexing and extracting the mixture of the sample to be detected and the supramolecular solvent in the centrifuge tube for 1 to 5 minutes, centrifuging for 3 to 8 minutes, removing the upper layer of liquid and blowing it with nitrogen, then re-dissolving it with acetonitrile solution, and filtering it through a filter membrane to obtain the extract to be detected.

[0014] Furthermore, the extract was vortexed for 3 min and centrifuged at 5000 r / min for 5 min.

[0015] Furthermore, a 10% acetonitrile solution and a 0.22 μm filter membrane were used.

[0016] Furthermore, in step 2, a series of standard solutions of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride with concentration gradients are first prepared, and the samples are detected by liquid chromatography-tandem mass spectrometry sample analysis method, and a chromatographic peak area-concentration standard correspondence relationship is established according to the detection results; the chromatographic peak areas of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be detected are determined by liquid chromatography-tandem mass spectrometry, and then the contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be detected are obtained according to the chromatographic peak area-concentration standard correspondence relationship.

[0017] Further, in step 2, liquid chromatography-tandem mass spectrometry is used for detection, and the chromatographic conditions are:

[0018] Chromatographic column: Waters Acquity UPLC BEH C18, 2.1 mm × 50 mm × 1.7 μm;

[0019] Gradient elution program: 0-5 min, 90%-60% 5 mmol / L ammonium acetate solution, 10%-40% acetonitrile; 5-7 min, 60%-30% 5 mmol / L ammonium acetate solution, 40%-70% acetonitrile; 7-8 min, 30% 5 mmol / L ammonium acetate solution, 70% acetonitrile; 8-8.1 min, 30%-90% 5 mmol / L ammonium acetate solution, 70%-10% acetonitrile; 8.1-10 min, 90% 5 mmol / L ammonium acetate solution, 10% acetonitrile;

[0020] Column temperature: 35°C;

[0021] Mobile phase flow rate: 0.5 mL / min;

[0022] Injection volume: 2 μL;

[0023] The mass spectrometry conditions were:

[0024] Ionization mode: electrospray ionization, positive ion;

[0025] Capillary voltage: 1.0kV;

[0026] Ion source temperature: 120°C;

[0027] Desolvation temperature: 450°C;

[0028] Desolventization gas flow rate: 650L / h;

[0029] Collision chamber pressure: 0.26MPa;

[0030] Cone hole back-blowing gas flow rate: 55L / h;

[0031] Scan mode: Multiple reaction monitoring.

[0032] The beneficial effects of the present invention are as follows: the present invention adopts a pretreatment technology of supramolecular solvent-dispersed liquid-liquid extraction, the pretreatment operation is simple and rapid, the cost is low, the enrichment efficiency is high, and the amount of organic solvent used is small; liquid chromatography-tandem mass spectrometry is used to simultaneously determine the approved hair dye tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in hair dye products, the specificity is good, the sensitivity is higher, and the sample analysis time is shortened, and the advantages of integrating extraction and purification, being green and environmentally friendly, etc. DETAILED DESCRIPTION

[0033] A preferred embodiment is given below to clearly and completely illustrate the present invention.

[0034] A method for detecting hair dye ingredients in a hair dye product comprises the following steps:

[0035] Step 1, weighing 0.5g of the sample to be tested into a centrifuge tube, adding a supramolecular solvent, wherein the supramolecular solvent is composed of 150μL of n-decanol, 750μL of tetrahydrofuran and 6mL of water; performing dispersed liquid-liquid extraction, that is, vortexing and extracting the mixture of the sample to be tested and the supramolecular solvent in the centrifuge tube for 3min, and centrifuging at 5000r / min for 5min; removing the upper liquid and blowing it dry with nitrogen, then re-dissolving it with 1mL of 10% acetonitrile solution, and filtering it through a 0.22μm filter membrane to obtain an extract to be tested.

[0036] Step 2, determining the contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be tested by liquid chromatography-tandem mass spectrometry.

[0037] In step 2, a series of standard solutions of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride with concentration gradients are first prepared, and the samples are detected by liquid chromatography-tandem mass spectrometry sample analysis method, and a chromatographic peak area-concentration standard correspondence relationship is established according to the detection results; the chromatographic peak areas of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be detected are determined by liquid chromatography-tandem mass spectrometry, and then the contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be detected are obtained according to the chromatographic peak area-concentration standard correspondence relationship.

[0038] In step 2, liquid chromatography-tandem mass spectrometry is used for detection, and the chromatographic conditions are:

[0039] Chromatographic column: Waters Acquity UPLC BEH C18, 2.1 mm × 50 mm × 1.7 μm;

[0040] Gradient elution program: 0-5 min, 90%-60% 5 mmol / L ammonium acetate solution, 10%-40% acetonitrile; 5-7 min, 60%-30% 5 mmol / L ammonium acetate solution, 40%-70% acetonitrile; 7-8 min, 30% 5 mmol / L ammonium acetate solution, 70% acetonitrile; 8-8.1 min, 30%-90% 5 mmol / L ammonium acetate solution, 70%-10% acetonitrile; 8.1-10 min, 90% 5 mmol / L ammonium acetate solution, 10% acetonitrile;

[0041] Column temperature: 35°C;

[0042] Mobile phase flow rate: 0.5 mL / min;

[0043] Injection volume: 2 μL;

[0044] The mass spectrometry conditions were:

[0045] Ionization mode: electrospray ionization, positive ion;

[0046] Capillary voltage: 1.0kV;

[0047] Ion source temperature: 120°C;

[0048] Desolvation temperature: 450°C;

[0049] Desolventization gas flow rate: 650L / h;

[0050] Collision chamber pressure: 0.26MPa;

[0051] Cone hole back-blowing gas flow rate: 55L / h;

[0052] Scan mode: Multiple reaction monitoring (MRM).

[0053] The qualitative / quantitative ion pairs, cone voltages, and collision energies are listed in Table 1.

[0054] Table 1 Qualitative / quantitative ion pairs, cone voltage and collision energy

[0055]

[0056] The concentrations of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride were linearly related to the chromatographic peak area in the range of 0.02-2 μg / mL, as shown in Table 2 for details.

[0057] Table 2 Linear data

[0058]

[0059] In Table 2, the R 2The R of 2,6-dimethoxy-3,5-pyridinediamine hydrochloride is 0.9993. 2 is 0.9996.

[0060] Using this detection method, the detection concentrations of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride were determined to be 0.005μg / mL based on the standard deviation of three and ten times the blank samples combined with the blank matrix sample spiked, and the minimum quantitative concentration was 0.02μg / mL. Based on 0.50g sample extracted and fixed to 10.0mL, the detection limit was 0.1mg / kg. The lower limit of quantification was 0.4mg / kg.

[0061] The sample extracts with different contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride were continuously injected and measured 6 times, and the samples with different contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride were extracted and measured 6 times according to the determined determination method. The calculated relative standard deviations (RSDs) were all less than 10%, indicating good precision and repeatability.

[0062] The sample extracts with different contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride were measured at 0h, 1h, 3h, 6h, 12h and 24h after sample preparation. The results showed that there was no significant difference in the values ​​and the stability was good within 24h.

[0063] The present invention adopts a pretreatment technology of supramolecular solvent-dispersed liquid-liquid extraction, and the pretreatment operation is simple and rapid, the cost is low, the enrichment efficiency is high, and the amount of organic solvent used is small; liquid chromatography-tandem mass spectrometry is used to simultaneously determine the approved hair dye tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in hair dye products, which has good specificity and higher sensitivity, and shortens the sample analysis time. It has the advantages of integrating extraction and purification, being green and environmentally friendly, and the like.

[0064] Although the specific embodiments of the present invention are described above, it should be understood by those skilled in the art that this is only for illustration and the protection scope of the present invention is defined by the appended claims. Those skilled in the art may make various changes or modifications to these embodiments without departing from the principles and essence of the present invention, but these changes and modifications all fall within the protection scope of the present invention.

Claims

1. A method for detecting hair dye ingredients in hair dye products, characterized in that: It includes the following steps: Step 1, adding a supramolecular solvent to a sample to be detected, and performing dispersed liquid-liquid extraction to obtain an extract to be detected; wherein the supramolecular solvent is composed of an alkyl alcohol, tetrahydrofuran and water; Step 2, determining the contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be tested by liquid chromatography-tandem mass spectrometry.

2. The method for detecting hair dye ingredients in hair dye products according to claim 1, characterized in that: In step 1, the alkyl alcohol in the supramolecular solvent is n-decanol.

3. The method for detecting hair dye ingredients in hair dye products according to claim 2, characterized in that: The ratio of n-decanol, tetrahydrofuran and water in the supramolecular solvent is 1:5:

40.

4. The method for detecting hair dye ingredients in hair dye products according to claim 1, characterized in that: In step 1, the dispersed liquid-liquid extraction step includes: vortexing and extracting the mixture of the sample to be detected and the supramolecular solvent in the centrifuge tube for 1 to 5 minutes, centrifuging for 3 to 8 minutes, removing the upper layer of liquid and blowing it to dryness with nitrogen, then re-dissolving it with acetonitrile solution, filtering it through a filter membrane, and obtaining the extract to be detected.

5. The method for detecting hair dye ingredients in hair dye products according to claim 4, characterized in that: The extract was vortexed for 3 min and centrifuged at 5000 rpm for 5 min.

6. The method for detecting hair dye ingredients in hair dye products according to claim 4, characterized in that: A 10% acetonitrile solution and a 0.22 μm filter membrane were used.

7. The method for detecting hair dye ingredients in hair dye products according to claim 1, characterized in that: In step 2, a series of standard solutions of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride with concentration gradients are first prepared, and the samples are detected by liquid chromatography-tandem mass spectrometry sample analysis method, and a chromatographic peak area-concentration standard correspondence relationship is established according to the detection results; the chromatographic peak areas of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be detected are determined by liquid chromatography-tandem mass spectrometry, and then the contents of tetraaminopyrimidine sulfate and 2,6-dimethoxy-3,5-pyridinediamine hydrochloride in the extract to be detected are obtained according to the chromatographic peak area-concentration standard correspondence relationship.

8. The method for detecting hair dye ingredients in hair dye products according to claim 1, characterized in that: In step 2, liquid chromatography-tandem mass spectrometry is used for detection, and the chromatographic conditions are: Chromatographic column: Waters Acquity UPLC BEH C18, 2.1 mm × 50 mm × 1.7 μm; Gradient elution program: 0-5 min, 90%-60% 5 mmol / L ammonium acetate solution, 10%-40% acetonitrile; 5-7 min, 60%-30% 5 mmol / L ammonium acetate solution, 40%-70% acetonitrile; 7-8 min, 30% 5 mmol / L ammonium acetate solution, 70% acetonitrile; 8-8.1 min, 30%-90% 5mmol / L ammonium acetate solution, 70%-10% acetonitrile; 8.1~10min, 90% 5mmol / L ammonium acetate solution, 10% acetonitrile; Column temperature: 35°C; Mobile phase flow rate: 0.5 mL / min; Injection volume: 2 μL; The mass spectrometry conditions were: Ionization mode: electrospray ionization, positive ion; Capillary voltage: 1.0kV; Ion source temperature: 120°C; Desolvation temperature: 450°C; Desolvation gas flow rate: 650L / h; Collision chamber pressure: 0.26MPa; Cone hole back-blowing gas flow rate: 55L / h; Scan mode: Multiple reaction monitoring.

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