A detection method for banned compounds in cosmetics based on QuEChERS combined with ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry

Abstract

The invention belongs to the technical field of cosmetics detection, and specifically discloses a detection method for banned compounds in cosmetics based on QuEChERS combined with ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry. The detection method comprises the following steps: S1, extraction: mixing a sample with an extraction solvent, ultrasonically extracting, and obtaining a supernatant containing the extract; S2, purification: taking the supernatant of step S1 into a centrifuge tube filled with a purifier, and then oscillating and centrifuging in sequence; then, filtering the supernatant after centrifugation through a membrane to obtain a purified liquid; S3, sample detection: using ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry to detect and analyze the purified liquid obtained in step S2. The invention provides a detection method for banned compounds in cosmetics, which realizes the detection of banned compounds in cosmetics, has the advantages of high sensitivity, accuracy and precision, and the pretreatment method is simple and fast.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics detection, and in particular relates to a method for detecting prohibited compounds in cosmetics based on QuEChERS combined with ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry. Background Art

[0002] With the rapid economic development and the improvement of living standards, the cosmetics market has flourished. While bringing huge economic benefits, the quality and safety of cosmetics have become the focus of people's attention. Few research institutions at home and abroad have carried out research on the detection of such banned drugs in cosmetics. Therefore, in order to effectively carry out the quality control and safety assessment of banned compounds, it is necessary to establish the determination method of the above drugs in cosmetics. Summary of the invention

[0003] In order to solve the above technical problems, the present invention provides a detection method for banned compounds in cosmetics based on QuEChERS combined with ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry. The detection method realizes the detection of banned compounds in cosmetics, has the advantages of high sensitivity, accuracy and precision, and the pretreatment method is simple and rapid, avoiding the problem of inaccurate detection of banned compounds due to the diversity and complexity of cosmetic matrices.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0005] A method for detecting banned compounds in cosmetics based on QuEChERS combined with ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry, the detection method comprising the following steps:

[0006] S1. Extraction: Mix the sample with the extraction solvent, perform ultrasonic extraction, and obtain a supernatant containing the extract;

[0007] S2, purification: take the supernatant of step S1 into a centrifuge tube filled with a purifier, and then perform oscillation and centrifugation in sequence; then filter the supernatant after centrifugation through a membrane to obtain a purified liquid;

[0008] S3. Sample detection: ultra high pressure liquid chromatography-triple quadrupole tandem mass spectrometry is used to detect and analyze the purified liquid obtained in step S2.

[0009] The prohibited compounds in the cosmetics are one or more of cyclophosphamide, azacyclohexanol, mepan, caramiphen hydrochloride, narcotine or metyrapone.

[0010] In the present invention, the QuEChERS rapid sample pretreatment technology is used for the detection of cyclophosphamide, azacyclohexanol, Wei An, caramifen hydrochloride, narcotine or metyrapone in cosmetics for the first time. The pretreatment process is simple and fast. The sample is extracted with a solvent and can be tested on the machine after rapid purification, which improves the extraction and purification efficiency of the sample. At the same time, an instrumental method for the simultaneous detection of the above 6 banned compounds is established for the first time, which fills the gap in compound detection technology and has the characteristics of high sensitivity, accuracy and precision. This technology can be used for risk assessment and screening monitoring of banned compounds in cosmetics, and has a broad application prospect.

[0011] Furthermore, when the sample in step S1 is specifically a water-based emulsion or essential oil cosmetic, the extraction step of step S1 is specifically as follows: accurately weigh (0.5-5) g of the sample into a 10 mL stoppered colorimetric tube, preferably 0.5 g, add an extraction solvent, oscillate for 0.5-1.5 min, preferably 1 min, to fully mix the sample and the extraction solvent, extract for 10-20 min, preferably 15 min, under 20-43 KHz, 200 W ultrasound, make up to 10 mL with the extraction solvent, shake well, and centrifuge at 10000 r / min for 5 min to obtain a supernatant containing the extract.

[0012] Furthermore, the sample in step S1 is specifically a water-based emulsion or essential oil cosmetic, and the extraction solvent in step S1 is one of water, methanol or acetonitrile, preferably methanol.

[0013] Commonly used extraction solvents for ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry include water, methanol, acetonitrile, etc. In the present invention, when the sample is an emulsion or essential oil cosmetic, methanol is selected as the extraction solvent, and the recovery rate of the six banned compounds meets the requirement of 80% to 120%.

[0014] Furthermore, when the sample in step S1 is specifically a cream cosmetic, the specific extraction step of step S1 is: accurately weigh (0.5-5) g of the sample into a 10 mL stoppered colorimetric tube, preferably 0.5 g, add a saturated sodium chloride solution to evenly disperse the sample, add an extraction solvent, oscillate for 0.5-1.5 min, preferably 1 min, to fully mix the sample and the extraction solvent, extract under 20-43 KHz, 200 W ultrasound for 10-20 min, preferably 15 min, make the volume up to 10 mL with the extraction solvent, shake well, and centrifuge at 10000 r / min for 5 min to obtain a supernatant containing the extract.

[0015] In the present invention, considering that the texture of cream-type cosmetics is relatively viscous, the use of extraction solvent alone may not be able to fully extract its substances, while the dispersion solvent has stronger penetration ability and will make the sample more uniform. The method of first using saturated sodium chloride solution as the dispersion solvent and then using solvent extraction can optimize the recovery rate of banned compounds.

[0016] Furthermore, the sample in step S2 is specifically a cream cosmetic, and the extraction solvent in step S1 is one of methanol, anhydrous ethanol or acetonitrile, preferably anhydrous ethanol.

[0017] In the present invention, when the sample is a cream cosmetic, anhydrous ethanol is selected as the extraction solvent, and the recovery rates of the six prohibited compounds meet the requirement of 80% to 120%.

[0018] Furthermore, the specific steps of purification in step S2 are: aspirate 1.00 mL of the supernatant of step S1 into a 1.5 mL centrifuge tube containing 30-110 mg of a purifier, preferably 50 mg of a purifier, oscillate for 2-4 min, preferably 2 min, centrifuge at 12000 r / min for 5 min, and pass the supernatant after centrifugation through a 0.22 μm organic filter membrane to obtain a purified liquid.

[0019] Furthermore, the purifier of the centrifuge tube in step S2 is neutral alumina (Al-N), aminopropyl powder (NH2), graphitized carbon black (GCB), N-propylethylenediamine (PSA), silica gel powder (Silica), or silica gel bonded octadecyl (C 18 ), preferably a silica-bonded octadecyl group (C 18 ).

[0020] In the present invention, when the sample is a lotion, essential oil or cream cosmetic, 50 mg of C 18 As a purifier, the recovery rates of the six banned compounds met the requirements of 80% to 120%.

[0021] Furthermore, the chromatographic conditions of the ultra-high performance liquid chromatography in step S3 are: chromatographic column: Waters BEH C 18 (100mm×2.1mm, 1.7μm); column temperature: 40℃; mobile phase: A is 0.1% (V / V) formic acid aqueous solution, B is acetonitrile; flow rate: 0.4mL / min; injection volume: 20μL; gradient elution program was used.

[0022] Furthermore, the mass spectrometry conditions in step S3 are: API 4000Q Trap UPLC-MS / MS (AB Company, USA), equipped with ESI ion source; positive ion scanning; MRM mode; curtain gas pressure: 15psi; nebulizer gas pressure: 50psi; heating auxiliary gas pressure: 50psi; collision gas pressure: MEDIUM; ionization pressure: 5500V; ion source temperature: 550°C.

[0023] Compared with the prior art, the present invention has achieved beneficial technical effects:

[0024] (1) The present invention is the first to use the QuEChERS rapid sample pretreatment technology for the detection of cyclophosphamide, azacyclohexanol, mepan, caramifen hydrochloride, narcotine, and metyrapone in cosmetics. The pretreatment process is simple and fast. The sample is extracted with a solvent and quickly purified before being tested on the machine, which improves the extraction and purification efficiency of the sample. At the same time, an instrumental method for the simultaneous detection of the above six banned compounds is established for the first time, filling the gap in compound detection technology and having the characteristics of high sensitivity, accuracy, and precision. This technology can be used for risk assessment and screening monitoring of banned compounds in cosmetics, and has a broad application prospect.

[0025] (2) Under the separation conditions adopted in the present invention, the six prohibited compounds in the present invention have different retention times and different quantitative and qualitative ions. When testing actual cosmetic samples, the retention time and ion abundance ratio of the suspected compounds in the cosmetics can be compared with those of the standard substances, thereby identifying whether the cosmetics contain the six prohibited compounds, eliminating mutual interference, achieving effective separation, and avoiding erroneous judgments.

[0026] (3) The present invention examines the effects of different variables on the recovery rates of six banned compounds and determines the optimal extraction process, thereby providing accurate detection technology for the six banned compounds and providing new detection ideas for other banned compounds in the future. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Standard curves for 6 banned compounds in cosmetics;

[0028] Figure 2 The chromatograms are of the banned compounds in 6 cosmetics. DETAILED DESCRIPTION

[0029] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. However, the scope of protection claimed in the present invention is not limited to the following specific embodiments, and the reagents or instruments used in the detection method of the present invention can be purchased from the market.

[0030] Example 1

[0031] This embodiment provides a detection method for determining prohibited compounds in cosmetics (the cosmetics in this embodiment are a kind of water-based emulsion or essential oil cosmetics) based on QuEChERS combined with ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry, comprising the following steps:

[0032] S1. Extraction: Accurately weigh 0.5 g (accurate to 0.001 g) of sample into a 10 mL stoppered colorimetric tube, add 8 mL of methanol, shake for 1 min to fully mix the sample and the extraction solvent, extract under 43 KHz, 200 W ultrasound for 15 min, make up to 10 mL with methanol, shake well, centrifuge at 10000 r / min for 5 min to obtain the supernatant containing the extract;

[0033] S2, purification: take 1.00 mL of the supernatant obtained in step S1, and add 50 mg C 18 The powder was placed in a 1.5 mL centrifuge tube, shaken for 2 min, centrifuged at 12000 r / min for 5 min, and then the supernatant was filtered through a 0.22 μm organic filter membrane to obtain a purified solution;

[0034] S3, sample detection: using ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry to detect and analyze the purified liquid obtained in step S2;

[0035] The chromatographic conditions of the ultra-high performance liquid chromatography of this embodiment are as follows: Chromatographic column: Waters BEH C 18 (100mm×2.1mm, 1.7μm); mobile phase: A is 0.1% (V / V) formic acid aqueous solution, B is acetonitrile, flow rate: 0.4mL / min; injection volume: 20μL; column temperature: 40℃; gradient elution program is shown in Table 1.

[0036] Mass spectrometry conditions: API 4000Q Trap UPLC-MS / MS (AB Company, USA), equipped with ESI ion source; positive ion scanning; MRM mode; curtain gas pressure: 15 psi; nebulizer gas pressure: 50 psi; heating auxiliary gas pressure: 50 psi; collision gas pressure: MEDIUM; ionization pressure: 5500 V; ion source temperature: 550°C.

[0037] Table 1 Chromatographic mobile phase gradient elution program

[0038] Time / min A B 0 85％ 15％ 6 20％ 80％ 9 20％ 80％ 12 50％ 50％ 16 85％ 15％ 17 85％ 15％

[0039] Example 2

[0040] This embodiment provides a detection method for determining prohibited compounds in cosmetics (the cosmetics in this embodiment are a type of cream) based on QuEChERS combined with ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry, comprising the following steps:

[0041] S1. Extraction: Accurately weigh 0.5 g (accurate to 0.001 g) of sample into a 10 mL stoppered colorimetric tube, add 1.0 mL of saturated sodium chloride solution to evenly disperse the sample, add 8 mL of anhydrous ethanol, shake for 1 min to fully mix the sample and the extraction solvent, extract under 43 KHz, 200 W ultrasound for 15 min, make up to 10 mL with anhydrous ethanol, shake well, and centrifuge at 10000 r / min for 5 min to obtain a supernatant containing the extract;

[0042] S2, purification: take 1.00 mL of the supernatant obtained in step S1, and add 50 mg C 18 The powder was placed in a 1.5 mL centrifuge tube, shaken for 2 min, centrifuged at 12000 r / min for 5 min, and then the supernatant was filtered through a 0.22 μm organic filter membrane to obtain a purified solution;

[0043] S3. Sample detection: ultra high pressure liquid chromatography-triple quadrupole tandem mass spectrometry is used to detect and analyze the purified liquid obtained in step S2.

[0044] The chromatographic conditions, mass spectrometry conditions and chromatographic mobile phase gradient elution procedure in this embodiment are the same as those in Example 1.

[0045] Example 3

[0046] This example investigates the influence of matrix effect on the recovery rate of 6 banned compounds.

[0047] 1.1 When the sample is a lotion or essential oil

[0048] (1) Take 100 μg / mL of the standard solution of the six banned compounds, place them in 10 mL volumetric flasks, dilute to volume with methanol, prepare 10 μg / mL single standard stock solution, and store in a refrigerator at -18°C. Take 1 mL of the 10 μg / mL single standard stock solution, place them in 10 mL volumetric flasks, dilute to volume with methanol, prepare 1 μg / mL mixed standard solution of the six banned compounds, and store in a refrigerator at -18°C.

[0049] (2) Using the stepwise dilution method, methanol and a blank aqueous emulsion or an essential oil cosmetic matrix extraction and purification solution (the extraction and purification solution is obtained by referring to steps S1 and S2 in Example 1) were used as constant volume solutions to dilute a 1 μg / mL mixed standard solution of six banned compounds, and two standard curves were drawn, with concentrations of 10, 20, 50, 100, and 200 ng / mL, respectively. Then, the two standard curves were used to quantitatively analyze the aqueous emulsion or essential oil samples spiked with a concentration of 20 ng / mL, and the recoveries of the six banned compounds were calculated. The results are shown in Table 2.

[0050] 1.2 When the sample is a cream

[0051] The stepwise dilution method was adopted, and anhydrous ethanol and an extraction and purification solution of a cream cosmetic matrix (the method for obtaining the extraction and purification solution refers to steps S1 and S2 in Example 2) were used as constant volume solutions to dilute 1 μg / mL of a mixed standard solution of six banned compounds (same as in 1.1), and two standard curves were drawn with concentrations of 10, 20, 50, 100, and 200 ng / mL, respectively. Then, the two standard curves were used to quantitatively analyze the cream samples spiked with a concentration of 20 ng / mL, and the recoveries of the six banned compounds were calculated respectively. The results are shown in Table 2.

[0052] The chromatographic conditions, mass spectrometry conditions and chromatographic mobile phase gradient elution procedure in this embodiment are the same as those in Example 1.

[0053] Table 2 Experimental results of matrix effect investigation

[0054]

[0055]

[0056] From the data in Table 2, it can be seen that when the samples are water emulsions and essential oils, the recovery rate is between 77.3% and 136.3% when the standard curve prepared with methanol is used for quantitative analysis of samples containing 6 banned compounds, and the recovery rate is between 82.5% and 116.0% when the standard curve prepared with blank matrix is ​​used for quantitative analysis, which meets the requirements of 80% to 120%. When the sample is a cream, when the standard curve prepared with anhydrous ethanol is used for quantitative analysis of samples containing 6 banned compounds, the recovery rates of cyclophosphamide and metyrapone are high, while when the curve prepared with blank matrix is ​​used for quantitative analysis, the recovery rate is between 92.8% and 110.3%, which meets the requirements of 80% to 120%. Therefore, the blank matrix is ​​determined to prepare the standard curve, so as to achieve the purpose of eliminating the matrix effect.

[0057] Example 4

[0058] This example investigates the effect of the type of purifier on the recovery rate of 6 banned compounds

[0059] The 1 μg / mL mixed standard solution of the six compounds was diluted to 0.02 μg / mL with methanol, and 1 mL of the 0.02 μg / mL mixed standard solution of the six compounds was added to the purifier silica gel bonded octadecyl (C 18 ), aminopropyl powder (NH2), graphitized carbon black (GCB), N-propylethylenediamine (PSA), silica gel powder (Silica), and neutral alumina (Al-N) were placed in 6 equal amounts of bullet-shaped centrifuge tubes, shaken and centrifuged, and detected by ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry, and the recoveries of the 6 banned compounds were calculated respectively, as shown in Table 3 for details.

[0060] Table 3 Effect of purifier types on the recovery rates of six banned compounds

[0061]

[0062] It can be seen from Table 3 that when C 18 When , PSA, and NH2 were used as purifiers, the recovery rates of the six banned compounds were concentrated between 80% and 120%; when AL-N was used as a purifier, the recovery rates of cyclophosphamide and azacyclohexanol were low, at 75.5% and 3.6%, respectively; when GCB was used as a purifier, the recovery rates of azacyclohexanol, caramifen hydrochloride, and metyrapone were low, among which the adsorption of azacyclohexanol was the greatest; when Silica was used as a purifier, the recovery rates of azacyclohexanol, caramifen hydrochloride, and metyrapone were low, and the recovery rate of narcotine was high. Since the main ingredients of lotions, creams, and essential oil cosmetics include some oils, lipids, etc., C 18 It mainly adsorbs non-polar compounds containing carbon chains, including fats, etc. PSA is effective in removing organic acids, pigments, and sugars in the matrix. NH2 mainly adsorbs some highly polar substances. Based on the characteristics of the three types of cosmetic ingredients, C was finally selected. 18 As its purifier.

[0063] Example 5

[0064] This example investigates the effect of the amount of purifier on the recovery of 6 banned compounds.

[0065] Referring to Example 3, the difference from Example 3 is that 200 μL of the mixed solution of the six compounds at 1 μg / mL is added to blank water emulsions, essential oils, and creams, and 50 mg of silica-bonded octadecyl (C 18 ) were replaced with 30 mg, 70 mg, 90 mg and 110 mg respectively. After being treated with S1 and S2, they were detected by ultra high pressure liquid chromatography-triple quadrupole tandem mass spectrometry, and the recoveries of the six banned compounds were calculated respectively. See Table 4 for details.

[0066] Table 4 Effect of the amount of purifier on the recovery rate of 6 banned compounds

[0067]

[0068]

[0069] As shown in Table 4, the recovery rates of the five purifiers in water-based emulsion cosmetics are all between 80% and 120%, with no significant difference; for essential oil cosmetics, C 18 At 50 mg, 70 mg, and 90 mg, the recovery rates were between 80% and 120%, and at 30 mg and 110 mg, the recovery rates of narcotine and azacyclohexanol were less than 80%. Therefore, considering the higher applicability of pretreatment and the wider application range, 50 mg C was selected based on the principle of economy. 18 As a purifier for lotions and essential oil cosmetics.

[0070] When the sample is a cream, the recovery rates of most compounds are between 80% and 120% at the five dosage levels. However, when the dosage is 30 mg, the recovery rates of each compound are between 114.0% and 120.8%, and the recovery rate of Wei An is slightly higher; when the dosage is 50 mg, the recovery rate is between 111.3% and 119.0%; when the dosage is 70 mg and above, the matrix effect becomes increasingly greater, and the recovery rate of some compounds is higher than 120%. Therefore, in order to take into account all compounds, 50 mg C is selected. 18 As a purifying agent for cream cosmetics.

[0071] Example 6

[0072] This example investigates the effect of sample weight on the recovery of 6 banned compounds

[0073] When the sample is an aqueous emulsion or essential oil cosmetic, refer to Example 3, except that 200 μL of a 1 μg / mL mixed solution of the six compounds was added to blank aqueous emulsions or essential oil cosmetics, and the sample weight was replaced with 1 g, 2 g, and 5 g. After being treated by S1 and S2, the sample was detected by ultra high pressure liquid chromatography-triple quadrupole tandem mass spectrometry, and the recoveries of the six banned compounds were calculated respectively, as shown in Table 5.

[0074] Table 5 Effect of sample weight on the recovery of 6 banned compounds

[0075]

[0076]

[0077] As shown in Table 5, when 0.5g of water-based cosmetics was weighed, the recovery rates of the six banned compounds ranged from 82.5% to 93.8%; when 1g of sample was weighed, the recovery rates ranged from 91% to 108.8%; when 2g of sample was weighed, the recovery rates ranged from 77.0% to 98.3%; when 5g of sample was weighed, the recovery rates ranged from 63.8% to 109.5%. It can be found that with the increase in the amount of sample weighed, the matrix effect increases. However, for essential oil cosmetics, when the sample weight was 1g, the recovery rates of cyclophosphamide, Wei An and Narcotic exceeded 120%. In order to make the pretreatment method more applicable, it was decided to weigh 0.5g of water-based and essential oil cosmetics.

[0078] Example 7

[0079] This example investigates the effect of extraction solvent on the recovery rate of 6 banned compounds

[0080] When the sample is an aqueous emulsion or essential oil cosmetic, refer to Example 3, except that 200 μL of a mixed solution of the six compounds at 1 μg / mL was added to a blank aqueous emulsion or essential oil cosmetic, and the extraction solvent was replaced with acetonitrile and water, while other conditions remained unchanged. After treatment with S1 and S2, the sample was detected by ultra-high pressure liquid chromatography-triple quadrupole tandem mass spectrometry, and the recoveries of the six banned compounds were calculated, as shown in Table 6.

[0081] Table 6 Effect of extraction solvent on the recovery of 6 banned compounds

[0082]

[0083]

[0084] From the results in Table 6, it can be seen that when methanol is used as the extraction solvent, the recovery rate of each compound is 82.5% to 116%; when acetonitrile is used as the extraction solvent, the recovery rate of each compound is 74.3% to 107.3%; when water is used as the extraction solvent, the recovery rate of each compound is 0% to 117.8%. Except for cyclophosphamide, the recovery rates of the other five compounds did not reach 80%, or even 0%. Therefore, acetonitrile and water are not suitable as common extraction solvents for water-based cosmetics and essential oils, so methanol is selected.

[0085] Example 8

[0086] This example investigates the effect of extraction solvent on the recovery rate of 6 banned compounds

[0087] When the sample is a cream cosmetic, refer to Example 3, except that 200 μL of a mixed solution of 6 compounds at 1 μg / mL is added to a blank cream cosmetic, and the extraction solvent is replaced with methanol, acetonitrile and anhydrous ethanol. After treatment with S1 and S2, ultra high pressure liquid chromatography-triple quadrupole tandem mass spectrometry is used for detection, and the recoveries of the 6 banned compounds are calculated respectively, as shown in Table 7 for details.

[0088] Table 7 Effect of extraction solvent on the recovery rate of 6 banned compounds

[0089]

[0090] As shown in Table 7, when methanol is used as the extraction solvent, the recovery rate is between 43.1% and 96.8%; when acetonitrile and anhydrous ethanol are used as the extraction solvent, the recovery rate of individual compounds is low or high, but when anhydrous ethanol is used for extraction, the response and peak shape of the substance are better than acetonitrile. Therefore, anhydrous ethanol is preferred for extraction. In addition, in order to optimize its recovery rate, considering that the texture of cream cosmetics is relatively viscous, the use of anhydrous ethanol extraction alone may not be able to fully extract its substances, and the dispersion solvent has stronger penetration ability, which will make the sample more uniform. Therefore, the use of saturated sodium chloride solution as a dispersion solvent and then anhydrous ethanol extraction is considered to improve its recovery rate. The results show that the recovery rate of this method is between 88.5% and 110.3%, which meets the requirements of a recovery rate of 80% to 120%.

[0091] Examples 9 and 10 are used to verify the accuracy, sensitivity and repeatability of the detection method of the present invention.

[0092] Example 9

[0093] Referring to Example 3, the difference from Example 3 is that the solution concentration (10, 20, 50, 100, 200 ng / mL) is used as the abscissa and the ion peak area of ​​the 6 prohibited compound pairs is used as the ordinate to draw a standard curve as shown in Figure 1 As shown. The results show that within the corresponding mass concentration range of the six banned compounds, the peak area of ​​the quantitative ion has a good linear relationship with the concentration, and the correlation coefficient R2 can reach above 0.99, which can meet the experimental analysis requirements. 100 μL of the 1 μg / mL mixed solution of the six compounds was added to blank emulsions, essential oils, and creams, respectively. After treatment with S1 and S2, the detection limit and quantification limit were calculated. The detection limit is usually 3 times the signal-to-noise ratio (S / N=3), and the quantification limit is usually 10 times the signal-to-noise ratio (S / N=10). See Table 8 for details.

[0094] Table 8 Linear equations, correlation coefficients and linear ranges of 6 banned compounds in cosmetics

[0095] Compound Linear equations Correlation coefficient LOD / mg / kg LOQ / mg / kg Cyclophosphamide y=5970x+4860 0.9995 0.01 0.02 Azacyclohexanol y=5670x-17300 0.9995 0.01 0.02 Stomach y=66700x-328000 0.9982 0.005 0.01 Caramiphene Hydrochloride y=2120x-3440 0.9997 0.05 0.1 Narcotic y=40100x+5580 0.9998 0.005 0.01 Metyrapone y=2000x-2850 0.9990 0.05 0.1

[0096] The chromatogram is as follows Figure 2 As shown in Table 9, the mass spectrometry parameters of 6 banned compounds in cosmetics were obtained through optimization.

[0097] Table 9 Retention time and mass spectrometry parameters of 6 banned compounds in cosmetics

[0098]

[0099]

[0100] * indicates quantitative ion

[0101] As can be seen from Table 9, the six prohibited compounds in the present invention have different retention times and different quantitative and qualitative ions. When testing actual cosmetic samples, the retention time and ion abundance ratio of the suspicious compounds in the cosmetics can be compared with the standard substances to identify whether the cosmetics contain the six prohibited compounds, thereby eliminating mutual interference, achieving effective separation, and avoiding erroneous judgments.

[0102] Example 10

[0103] Referring to the method of preparing and detecting the blank matrix spiked sample solution in Example 4, the blank matrix was spiked with banned compounds at three concentration levels, and six groups of parallel tests were performed simultaneously. The recoveries (%) and relative standard deviations (RSDs) of the three spiked concentration levels and the test results are shown in Table 10.

[0104] Table 10 Spiked recovery (n=6) and relative standard deviation of three types of cosmetics at different concentration levels

[0105]

[0106]

[0107] It can be seen from Table 10 that when the samples are lotions and essential oil cosmetics, the recoveries of the six banned compounds are between 92.2% and 115.2%, and the relative standard deviations (RSD) are between 1.96% and 13.09%; when the samples are cream cosmetics, the recoveries of the six banned compounds are between 90.1% and 113.9%, and the RSD is between 2.20% and 13.10%. Both the recovery and precision meet the requirements of GB / T27040-2008.

[0108] Test Example 1

[0109] Different categories of cosmetics were collected, including 22 lotions and 7 essential oils. The method of Example 1 was used for testing, and no six banned compounds were detected. Different categories of cosmetics were collected, including 18 creams and ointments. The method of Example 2 was used for testing, and no six banned compounds were detected.

[0110] According to the disclosure and teaching of the above description, those skilled in the art to which the present invention belongs may also make changes and modifications to the above embodiments. Therefore, the present invention is not limited to the specific embodiments disclosed and described above, and some modifications and changes to the invention should also fall within the scope of protection of the claims of the present invention. In addition, although some specific terms are used in this specification, these terms are only for the convenience of description and do not constitute any limitation to the invention.

Claims

1. A method for detecting banned compounds in cosmetics based on QuEChERS combined with ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry, characterized in that: The detection method comprises the following steps: S1. Extraction: Mix the sample with the extraction solvent, perform ultrasonic extraction, and obtain a supernatant containing the extract; S2, purification: taking the supernatant of step S1 into a centrifuge tube filled with a purifier, and then oscillating and centrifuging in sequence, and then filtering the supernatant after centrifugation through a membrane to obtain a purified liquid; S3, sample detection: ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry is used to detect and analyze the purified liquid obtained in step S2. The banned compounds in the cosmetics are cyclophosphamide, azacyclohexanol, mepan, caramifen hydrochloride, narcotine and metyrapone, and the purifier is silica gel bonded octadecyl. The chromatographic conditions of the ultra-high performance liquid chromatography are as follows: Chromatographic column: Waters BEH C 18 ; Column temperature: 40°C; Mobile phase: A is 0.1% (V / V) formic acid aqueous solution, B is acetonitrile; Flow rate: 0.4mL / min; Injection volume: 20μL; Use gradient elution program, The gradient elution program is shown in the following table: The mass spectrometry conditions are: API 4000Q Trap UPLC-MS / MS, equipped with ESI ion source; positive ion scan; MRM mode; curtain gas pressure: 15 psi; nebulizer gas pressure: 50 psi; heating auxiliary gas pressure: 50 psi; collision gas pressure: MEDIUM; ionization pressure: 5500 V; ion source temperature: 550°C.

2. The detection method according to claim 1, characterized in that: The sample in step S1 is specifically a water-based emulsion or essential oil cosmetic. The specific extraction steps of step S1 are: accurately weigh (0.5-5) g of the sample into a 10 mL stoppered colorimetric tube, add an extraction solvent, oscillate for 0.5-1.5 min to fully mix the sample and the extraction solvent, extract for 10-20 min under 20~43KHz, 200W ultrasound, make the volume up to 10 mL with the extraction solvent, shake well, and centrifuge at 10000r / min for 5 min to obtain a supernatant containing the extract.

3. The detection method according to claim 2, characterized in that: The extraction solvent in step S1 is one of water, methanol or acetonitrile.

4. The detection method according to claim 1, characterized in that: The sample in step S1 is specifically a cream cosmetic, and the specific extraction steps of step S1 are: accurately weigh (0.5-5) g of the sample into a 10 mL stoppered colorimetric tube, add a saturated sodium chloride solution to evenly disperse the sample, add an extraction solvent, oscillate for 0.5-1.5 minutes to fully mix the sample and the extraction solvent, extract for 10-20 minutes under 20~43KHz, 200W ultrasound, make the volume up to 10 mL with the extraction solvent, shake well, and centrifuge at 10000r / min for 5 minutes to obtain a supernatant containing the extract.

5. The detection method according to claim 4, characterized in that: The extraction solvent in step S1 is one of methanol, anhydrous ethanol or acetonitrile.

6. The detection method according to claim 1, characterized in that: The specific steps of purification in step S2 are: 1.00 mL of the supernatant of step S1 is taken into a 1.5 mL centrifuge tube containing 30-110 mg of a purifier, shaken for 2-4 minutes, centrifuged at 12000 r / min for 5 minutes, and the supernatant after centrifugation is filtered through a 0.22 μm organic filter membrane to obtain a purified solution.

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