阅读说明：本技术 同时检测化妆品中酰胺类及吡咯烷酮类有机溶剂残留量的方法 (Method for simultaneously detecting residual quantity of amide and pyrrolidone organic solvents in cosmetics ) 是由 丁友超 侯建军 费晓庆 吴斌 汤娟 周佳 钱凯 张普霞 于 2021-08-30 设计创作，主要内容包括：本发明涉及工业产品的检测领域,特别是涉及化妆品类的检测领域,更为具体的说是涉及一种同时检测化妆品中酰胺类及吡咯烷酮类有机溶剂残留量的方法,通过气相色谱-质谱联用的检测方法,建立了通过内标法测定化妆品中10种酰胺类和吡咯烷酮类有机溶剂的定量检测方法。本发明公开的方法具有分析物种类多、分离效果佳、稳定性好的优势,具有很高的实际应用价值,可以为我国相关部门的监管工作提供科学依据和数据基础,为制定相关行业标准及国家标准提供了参考,对保护消费者利益起到重要作用。(The invention relates to the field of detection of industrial products, in particular to the field of detection of cosmetics, and more particularly relates to a method for simultaneously detecting residual amounts of amide and pyrrolidone organic solvents in cosmetics. The method disclosed by the invention has the advantages of multiple analyte types, good separation effect and good stability, has high practical application value, can provide scientific basis and data basis for the supervision work of relevant departments in China, provides reference for formulating relevant industrial standards and national standards, and plays an important role in protecting the benefits of consumers.)

1. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics is characterized by comprising the following steps: the method is a method for quantitatively detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics by adopting a gas chromatography-mass spectrometry combined detection system and taking mixed internal standard working solution formed by mixing an internal standard substance of an amide compound and an internal standard substance of a pyrrolidone compound as an internal standard reference substance.

2. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics according to claim 1, wherein any one or more of the following conditions are preferred:

(1) the temperature rising procedure of the gas chromatography is as follows: the initial temperature is 50 deg.C, keeping for 1min, heating to 120 deg.C at a speed of 10 deg.C/min, keeping for 2min, heating to 200 deg.C at a speed of 10 deg.C/min, and keeping for 5 min;

(2) the detection conditions of the mass spectrum are as follows: an Electron Impact (EI) ion source; ionization energy is 70 eV; the ion source temperature is 230 ℃; the temperature of the four-level bar is 150 ℃; the temperature of the transmission line is 280 ℃; a full SCAN (SCAN) mode; the scanning range is (20-300) amu; the solvent delay time is 6 min;

(3) the gas chromatography adopts an ultrahigh inert capillary column as a chromatographic column, the carrier gas adopts high-purity helium, and the flow rate of the carrier gas is 1.0 mL/min.

3. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics according to claim 1, wherein the internal standard substance of the amide compound is N, N-diethylformamide, and the internal standard substance of the pyrrolidone compound is N-tetradecane.

Further preferably, the concentration of N, N-diethylformamide in the mixed internal standard working solution is 1000mg/L, and the concentration of N-tetradecane is 500 mg/L.

4. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics according to claim 1, further comprising a pretreatment step, wherein the pretreatment methods of different samples respectively comprise:

a. accurately weighing a powdery sample into a centrifuge tube, adding a mixed internal standard working solution and ethyl acetate, carrying out ultrasonic extraction at normal temperature, centrifuging, and absorbing supernatant as a sample to be detected;

b. accurately weighing an oily sample, a pasty sample and an emulsion sample in a centrifuge tube, adding a mixed internal standard working solution and a saturated sodium chloride aqueous solution, performing ultrasonic dispersion at normal temperature, adding ethyl acetate, performing oscillation extraction and centrifugation, and absorbing supernatant as a sample to be detected;

c. and for a liquid water-based sample, accurately weighing the sample in a centrifuge tube, adding mixed internal standard working solution, sodium chloride powder and ethyl acetate, oscillating, extracting, centrifuging, adding anhydrous sodium sulfate, and absorbing supernatant to serve as a sample to be detected.

5. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics according to claim 4, wherein the addition ratio of the volume of the extraction solution to the mixed internal standard working solution is 10 mL: 100 μ L.

6. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics according to claim 4, wherein the oscillation frequency of the oscillation extraction is 150 r/min. The shaking time is preferably 30 min.

7. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics according to claim 4, wherein the centrifugation speed is 8000r/min, preferably the centrifugation time is 5 min.

8. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics according to claim 1, further comprising the step of drawing a quantitative curve, comprising the following steps:

firstly, 10 kinds of standard substance stock solutions of analytes to be detected are prepared respectively,

secondly, quantitatively transferring a certain volume of standard product stock solution, and preparing to form mixed standard working solution;

step three, diluting the mixed standard working solution step by step, and adding the mixed internal standard working solution;

and fourthly, measuring by adopting a gas chromatography-mass spectrometry combined detection system, taking the concentration of each analyte as a horizontal coordinate, taking the ratio of the peak area of each analyte to the peak area of the corresponding internal standard substance as a vertical coordinate, and respectively establishing 10 standard curves of the analytes to be detected.

9. The method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics according to claim 8, wherein the concentrations of 10 analyte standards to be detected in the mixed standard working solution are 200mg/L, N formamide, 100mg/L, N N-dimethylformamide, 100mg/L, N N-methylacetamide, 100mg/L, N N-dimethylacetamide and 100mg/L, N thioacetamide, 100mg/L, N N-vinylpyrrolidone and 100mg/L, N Ethyl pyrrolidone respectively.

Technical Field

The invention relates to the field of detection of industrial products, in particular to the field of detection of cosmetics, and more particularly relates to a method for simultaneously detecting residual amounts of amide and pyrrolidone organic solvents in cosmetics.

Background

Amide and pyrrolidone organic solvents can dissolve and disperse essence, sterilization preservative, surfactant, grease, colorant and the like, so that the organic solvents are widely used in the preparation process of industrial products such as cosmetics, textiles and the like. Since the 70 th 20 th century, along with the extensive studies on the toxicity of these two organic solvents at home and abroad, carcinogenic, teratogenic or reproductive toxicity has been discovered, so that various legislations in various countries and regions limit the use of these organic solvents, such as: european union chemical registration, evaluation, approval, and Restriction (REACH) regulations place 7 organic solvents, such as N-methylpyrrolidone, N-ethylpyrrolidone, N-methylformamide, N-methylacetamide, N-dimethylformamide, N-dimethylacetamide, and formamide, in restricted materials list appendix 17; the ecological textile Standard Oeko-Tex Standard 100, version 2021, sets forth limited requirements for formamide, N-methylpyrrolidone, N-dimethylformamide and N, N-dimethylacetamide; the European chemical administration (ECHA) lists 5 organic solvents such as N-methylpyrrolidone, N-methylacetamide, N-dimethylformamide, N-dimethylacetamide and formamide as highly interesting Substances (SVHC); the Washington State Children's Product Safety Act (CPSA) lists N-methylpyrrolidone as a substance of high interest to children (CHCC); the year 2017, the month 10 and the day 27,WORLD HEALTH ORGANIZATIONthe international agency for research on cancer places thioacetamide on the list of class 2B carcinogens; safety technical Specification for cosmetics (2)015 edition) formamide, acetamide, N-dimethylformamide, N-dimethylacetamide, N-methylformamide, N-methylacetamide, thioacetamide and N-vinylpyrrolidone are listed as forbidden components.

At present, detection of amide and pyrrolidone organic solvents of products such as textiles, leather and the like is reported, but detection of residual amounts of amide and pyrrolidone organic solvents in cosmetic products is not reported yet.

Different from the detection of textiles, leather and the like, the cosmetic products have various industrial product forms, including powder products, paste products, emulsion products, oily products, liquid water-based products and the like, so the universality of the method is ensured, and the method can be suitable for different product forms and is one of the technical problems to be solved in the detection of the cosmetic products. Meanwhile, the residual organic solvent in the cosmetic products is not easy to extract and separate, so how to ensure that the residual amide and pyrrolidone organic solvents in the products are effectively extracted and further ensure the reliability of detection results is also a technical problem to be solved in the detection of the cosmetic products. Finally, the types of amides and pyrrolidones which may be contained in the cosmetic products are various, and the common types include 10, so how to detect the residual quantity of the 10 amides and pyrrolidones organic solvents at one time is also a technical problem which needs to be overcome in the detection of the cosmetic industrial products.

Disclosure of Invention

The invention aims to solve the technical problem of how to accurately detect the residual quantity of amide and pyrrolidone organic solvents in cosmetics.

In order to solve the technical problems, the invention discloses a method for simultaneously detecting the residual quantity of amide and pyrrolidone organic solvents in cosmetics, which is a method for quantitatively detecting the residual quantity of the amide and pyrrolidone organic solvents in cosmetics by adopting a detection system combining gas chromatography and mass spectrometry and taking mixed internal standard working solution formed by mixing an internal standard substance of an amide compound and an internal standard substance of a pyrrolidone compound as an internal standard reference substance.

The amide organic solvent in the present invention means formamide, N-dimethylformamide, N-methylformamide, acetamide, N-methylacetamide, N-dimethylacetamide, thioacetamide; the pyrrolidone includes N-methyl pyrrolidone, N-vinyl pyrrolidone and N-ethyl pyrrolidone.

The gas chromatography adopts an ultrahigh inert capillary column as a chromatographic column, the carrier gas adopts high-purity helium, and the flow rate of the carrier gas is 1.0 mL/min.

The temperature rising procedure of the gas chromatography is as follows: the initial temperature is 50 deg.C, holding for 1min, heating to 120 deg.C at a speed of 10 deg.C/min, holding for 2min, heating to 200 deg.C at a speed of 10 deg.C/min, and holding for 5 min.

The detection conditions of the mass spectrum are as follows: an Electron Impact (EI) ion source; ionization energy is 70 eV; the ion source temperature is 230 ℃; the temperature of the four-level bar is 150 ℃; the temperature of the transmission line is 280 ℃; a full SCAN (SCAN) mode; the scanning range is (20-300) amu; the solvent delay time was 6 min.

Further preferably, the internal standard substance of the amide compound is N, N-diethylformamide, and the internal standard substance of the pyrrolidone compound is N-tetradecane.

Further preferably, the concentration of N, N-diethylformamide in the mixed internal standard working solution is 1000mg/L, and the concentration of N-tetradecane is 500 mg/L.

Further preferably, the invention also discloses pretreatment methods of different samples, which respectively comprise the following steps:

a. accurately weighing a powdery sample into a centrifuge tube, adding a mixed internal standard working solution and ethyl acetate, carrying out ultrasonic extraction at normal temperature, centrifuging, and absorbing supernatant as a sample to be detected;

b. accurately weighing an oily sample, a pasty sample and an emulsion sample in a centrifuge tube, adding a mixed internal standard working solution and a saturated sodium chloride aqueous solution, performing ultrasonic dispersion at normal temperature, adding ethyl acetate, performing oscillation extraction and centrifugation, and absorbing supernatant as a sample to be detected;

c. and for a liquid water-based sample, accurately weighing the sample in a centrifuge tube, adding mixed internal standard working solution, sodium chloride powder and ethyl acetate, oscillating, extracting, centrifuging, adding anhydrous sodium sulfate, and absorbing supernatant to serve as a sample to be detected.

In a preferred technical scheme, the adding ratio of the volume of the extraction solution to the mixed internal standard working solution is 10 mL: 100 μ L.

Further preferably, the oscillation frequency of the oscillation extraction is 150 r/min. The shaking time is preferably 30 minutes.

Further preferably, the rotation speed of the centrifugation is 8000r/min, and the centrifugation time is 5 min.

In a preferred technical scheme, the method also comprises the step of drawing a quantitative curve, and the method comprises the following steps:

firstly, 10 kinds of standard substance stock solutions of analytes to be detected are prepared respectively,

secondly, quantitatively transferring a certain volume of standard product stock solution, and preparing to form mixed standard working solution;

step three, diluting the mixed standard working solution step by step, and adding the mixed internal standard working solution;

and fourthly, measuring by adopting a gas chromatography-mass spectrometry combined detection system, taking the concentration of each analyte as a horizontal coordinate, taking the ratio of the peak area of each analyte to the peak area of the corresponding internal standard substance as a vertical coordinate, and respectively establishing 10 standard curves of the analytes to be detected.

Further preferably, the concentrations of 10 analyte standards to be detected in the mixed standard working solution are 200mg/L, N formamide, 100mg/L, N N-dimethylformamide-100 mg/L N-dimethylformamide, 100mg/L, N acetamide-100 mg/L, N methylacetamide, 100mg/L N-dimethylacetamide, 100mg/L, N thioacetamide, 100mg/L, N-vinylpyrrolidone, 100mg/L, N-ethylpyrrolidone and 100mg/L N-dimethylacetamide respectively.

The quantitative detection method of 10 amide and pyrrolidone organic solvents in cosmetics is established by a gas chromatography-mass spectrometry combined detection method, and through the detection of an addition recovery experiment, the average recovery rate of 10 analytes in oily, powdery, cream, emulsion and liquid water-based samples is 80.5-104.6% within the addition level range of (0.5-50) mg/kg, and the RSD value is not more than 7.8%. The quantitative limit of other analytes in 10 analytes is 0.1mg/L and the linear range is 0.1-50mg/L except that the quantitative limit of formamide is 0.2mg/L and the linear range is 0.2-100 mg/L. The method disclosed by the invention has the advantages of multiple analyte types, good separation effect and good stability, has high practical application value, can provide scientific basis and data basis for the supervision work of relevant departments in China, provides reference for formulating relevant industrial standards and national standards, and plays an important role in protecting the benefits of consumers.

Drawings

FIG. 1 is a total ion flow chromatogram of gas chromatography-mass spectrometry of 10 amide and pyrrolidone organic solvent standard substances.

FIG. 2 is the GC-MS total ion current chromatogram of the cationic emulsion sample in example 2.

Detailed Description

In order that the invention may be better understood, we now provide further explanation of the invention with reference to specific examples.

EXAMPLE 110 Standard Curve plotting of amide-based and pyrrolidone-based organic solvents

Respectively weighing a certain amount of 10 analyte standard substances to be detected, dissolving the 10 analyte standard substances with ethyl acetate, fixing the volume to 10mL, preparing a single standard stock solution with the mass concentration of 1000mg/L, respectively transferring a certain volume of the standard stock solution, and preparing a mixed standard working solution, wherein the concentration of formamide is 200mg/L, and the rest concentrations are 100 mg/L.

Respectively weighing a certain amount of two internal standard substances N, N-diethylformamide and N-tetradecane, dissolving by using ethyl acetate, fixing the volume to 10mL, preparing a single internal standard stock solution with the mass concentration of 5000mg/L, and then respectively transferring a certain volume of the internal standard stock solution to prepare a mixed internal standard working solution, wherein the concentration of the N, N-diethylformamide is 1000mg/L, and the concentration of the N-tetradecane is 500 mg/L.

And diluting the mixed standard working solution step by step into a series of standard working solutions (the concentration of the mixed internal standard working solution contained in each standard working point is the same as that of the sample pretreatment part), and measuring according to optimized instrument conditions.

Gas chromatography conditions: the chromatographic column was a DB-624 ultra-high inert capillary column (30 m.times.0.25 mm,1.4 μm). Temperature rising procedure: the initial temperature is 50 deg.C, holding for 1min, heating to 120 deg.C at a speed of 10 deg.C/min, holding for 2min, heating to 200 deg.C at a speed of 10 deg.C/min, and holding for 5 min. The temperature of a sample inlet is 200 ℃; the carrier gas is high-purity helium (99.999%); the flow rate is 1.0 mL/min; no shunt sampling; the amount of sample was 1.0. mu.L.

Mass spectrum conditions: an Electron Impact (EI) ion source; ionization energy is 70 eV; the ion source temperature is 230 ℃; the temperature of the four-level bar is 150 ℃; the temperature of the transmission line is 280 ℃; a full SCAN (SCAN) mode; the scanning range is (20-300) amu; the solvent delay time was 6 min.

The results are shown in table 1 and figure 1,

table 1:

serial number	Name (R)	CAS No.	Quantitative ion	Qualitative ion	Abundance ratio	Retention time/min
							1	Carboxamides	75-12-7	45	45:44:43:29	100:30:24:20	7.18
2	N, N-dimethylformamide	68-12-2	73	73:44:42:28	100:49:20:12	7.83
							3	N-methylformamide	123-39-7	59	59:28:30:58	100:25:23:12	8.15
4	Acetamide	60-35-5	59	59:44:43:42	100:59:41:19	8.33
							5	N-methylacetamide	79-16-3	73	73:43:58:30	100:68:58:30	9.38
6	N, N-dimethyl acetamide	127-19-5	87	87:44:43:72	100:69:32:14	9.70
							7	N-methyl pyrrolidone	872-50-4	99	99:98:44:42	100:84:42:31	14.01
8	Thioacetamide	62-55-5	75	75:60:59:42	100:37:18:16	14.78
							9	N-vinyl pyrrolidone	88-12-0	111	111:56:28:82	100:98:17:16	15.16
10	N-ethylpyrrolidone	2687-91-4	113	113:98:70:56	100:80:30:22	15.23

Taking N, N-diethylformamide as an internal standard of an amide compound and marking the internal standard as 1, taking N-tetradecane as an internal standard of a pyrrolidone compound and marking the internal standard as 2, wherein mass spectrum analysis parameters are shown in a table 2:

table 2:

using an internal standard method for quantification, taking the concentration of each analyte as an abscissa (x), taking the ratio of the peak area of 10 analytes to the peak area of the corresponding internal standard as an ordinate (y), establishing a standard curve to obtain a corresponding linear regression equation, calculating the quantification Limit (LOQ) of the instrument by using a 10-fold signal-to-noise ratio (S/N ═ 10), and obtaining detailed results shown in Table 3,

TABLE 3

Example 2

Firstly, selecting a pretreatment method according to different sample forms, wherein in the embodiment, the sample is emulsion, so that the pretreatment is carried out according to a pretreatment method b, namely, accurately weighing the sample in a centrifuge tube, adding a mixed internal standard working solution and a saturated sodium chloride aqueous solution, carrying out normal-temperature ultrasonic dispersion, adding ethyl acetate, carrying out oscillation extraction and centrifugation, and absorbing supernatant as a sample to be detected;

in this example, 2.0g (accurate to 0.01g) of sample is weighed into a 50mL centrifuge tube with a cover, 100 μ L of mixed internal standard working solution and 10mL of saturated sodium chloride aqueous solution are added, after the sample is dispersed by ultrasound, 10mL of ethyl acetate is added, the cover is tightly covered, after oscillation extraction is carried out for 30min at the oscillation frequency of 150r/min, 8000r/min is centrifuged for 5min, a proper amount of anhydrous sodium sulfate is added, and a part of upper clear liquid is absorbed and filtered to a sample injection bottle through an organic phase needle type filter head, and is used for gas chromatography-mass spectrometer measurement.

The measurements were carried out according to the instrument conditions disclosed in example 1, in particular:

gas chromatography conditions: the chromatographic column was a DB-624 ultra-high inert capillary column (30 m.times.0.25 mm,1.4 μm). Temperature rising procedure: the initial temperature is 50 deg.C, holding for 1min, heating to 120 deg.C at a speed of 10 deg.C/min, holding for 2min, heating to 200 deg.C at a speed of 10 deg.C/min, and holding for 5 min. The temperature of a sample inlet is 200 ℃; the carrier gas is high-purity helium (99.999%); the flow rate is 1.0 mL/min; no shunt sampling; the amount of sample was 1.0. mu.L.

Mass spectrum conditions: an Electron Impact (EI) ion source; ionization energy is 70 eV; the ion source temperature is 230 ℃; the temperature of the four-level bar is 150 ℃; the temperature of the transmission line is 280 ℃; a full SCAN (SCAN) mode; the scanning range is (20-300) amu; the solvent delay time was 6 min.

The results of the detection are shown in FIG. 2.

Wherein the peak area of the compound 4, namely acetamide, is 177767410, the peak area of the internal standard 1 is 72301849,according to the corresponding linear equation in table 2: y is 8.024 × 10^-2x+2.868×10^-3X was found to be 30.61 mg/L. Therefore, the emulsion contained acetamide, and the content of acetamide was 153.05 mg/kg.

What has been described above is a specific embodiment of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and such improvements and modifications are also considered to be within the scope of the present invention.