CN105067746A - Method for determination of padimate in cosmetics - Google Patents
Method for determination of padimate in cosmetics Download PDFInfo
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- CN105067746A CN105067746A CN201510475211.5A CN201510475211A CN105067746A CN 105067746 A CN105067746 A CN 105067746A CN 201510475211 A CN201510475211 A CN 201510475211A CN 105067746 A CN105067746 A CN 105067746A
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Abstract
The invention discloses a method for determination of padimate in cosmetics, wherein the method includes the following steps: (1) pretreating samples; (2) analyzing padimate by high performance liquid chromatography, and optimizing high performance liquid chromatography conditions; and (3) drawing a standard working curve, quantifying by adopting an external standard method, and calculating the content of padimate. The method for determination of padimate in the cosmetics adopts the high performance liquid chromatography for simultaneous determination of padimate A and padimate O in the cosmetics; the samples detected to be positive by the high performance liquid chromatography are further identified by using high performance liquid chromatography-mass spectrometry/mass spectrometry. The method is accurate and fast, is high in sensitivity, and has a great significance for monitoring of the cosmetics quality safety risks.
Description
Technical field
The present invention relates to a kind of assay method of chemical substance, particularly relate to the assay method of padimate in cosmetics.
Background technology
Padimate is divided into Padimate A and Padimate O.Padimate A, relative molecular mass: 235.36, CAS:14779-78-3, molecular formula: C
14h
21nO
2, structural formula is as follows:
Padimate O, relative molecular mass: 277.41, CAS:21245-02-3, molecular formula: C
17h
27nO
2, structural formula is as follows:
Padimate A has spread effect due to it to eyes, respiratory tract and skin, Padimate A must not be added in clear stipulaties cosmetics in China's " cosmetics health specification ", Padimate O is as the anti-ultraviolet absorbing agent of one, be widely used in cosmetics and the anti-Exposure to Sunlight industrial goods of other printing opacity, limitation is 8%.Report is had no to the detection method of padimate in cosmetics both at home and abroad.
Summary of the invention
The technical problem to be solved in the present invention is to provide the assay method of padimate in a kind of accurate, quick, highly sensitive cosmetics.
The assay method of padimate in cosmetics, it comprises the steps:
(1) pre-treatment of sample;
(2) adopt the method for high performance liquid chromatography to analyze padimate, optimize high-efficient liquid phase chromatogram condition;
(3) drawing standard working curve, adopts quantified by external standard method, calculates the content of padimate.
The assay method of padimate in cosmetics of the present invention, wherein, described high-efficient liquid phase chromatogram condition is as follows:
Chromatographic column: XTerraC
18(4.6mm × 250mm, 5 μm);
Flow velocity: 1.0mL/min;
Mobile phase: acetonitrile-water mixed solution, wherein the volume ratio of acetonitrile and water is 9:1, isocratic elution;
Determined wavelength: 310nm;
Column temperature: 30 DEG C;
Sample size: 10 μ L.
The assay method of padimate in cosmetics of the present invention, wherein, described sample is cream kind, lipstick class, aqua class, loose powder class or shampoo class cosmetic sample.
The assay method of padimate in cosmetics of the present invention, wherein, described sample is cream kind cosmetic sample, and pre-treatment comprises the steps: that getting 1g sample is placed in 10mL color comparison tube, after adding 2g sodium chloride breakdown of emulsion, add methyl alcohol to scale, fully mix, ultrasonic assistant extracts 20min, be transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, get supernatant and cross film, filtrate is for high-performance liquid chromatogram determination.
The assay method of padimate in cosmetics of the present invention, wherein, described sample is lipstick class cosmetic sample, and pre-treatment comprises the steps: that getting 1g sample is placed in 10mL color comparison tube, add 2mL tetrahydrofuran, add methyl alcohol to scale, fully mix, ultrasonic assistant extracts 20min, be transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, get supernatant and cross film, filtrate is for high-performance liquid chromatogram determination.
The assay method of padimate in cosmetics of the present invention, wherein, described sample is aqua class, loose powder class or shampoo class cosmetic sample, and pre-treatment comprises the steps: that getting 1g sample is placed in 10mL color comparison tube, adds methyl alcohol to scale, abundant mixing, ultrasonic assistant extracts 20min, is transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, get supernatant and cross film, filtrate is for high-performance liquid chromatogram determination.
The assay method of padimate in cosmetics of the present invention, wherein, for adopting high performance liquid chromatography to detect positive sample, adopts High Performance Liquid Chromatography/Mass Spectrometry/mass spectroscopy to confirm further, wherein:
High-efficient liquid phase chromatogram condition is as follows:
Chromatographic column: HSST3 chromatographic column (100mm × 2.1mm, 1.8 μm);
Mobile phase: volume ratio is 0.1% aqueous formic acid and the methyl alcohol of 1:9;
Flow velocity: 0.4mL/min;
Column temperature: 30 DEG C;
Sample size: 10 μ L;
Mass Spectrometry Conditions is as follows:
Ion gun: electric spray ion source;
Ionization mode: positive ion mode;
Capillary voltage: 3.5kV;
Extraction voltage: 2.9V;
Ion source temperature: 150 DEG C;
Desolventizing temperature degree: 350 DEG C;
Data acquisition modes: many reactive ion monitorings.
The assay method of padimate in cosmetics of the present invention, wherein, described padimate is Padimate A and Padimate O.
The assay method of padimate in cosmetics of the present invention, wherein, described confirmatory test comprises the steps: sample liquid suitably to dilute, sample liquid and standard working solution is measured by High Performance Liquid Chromatography/Mass Spectrometry/Mass Spectrometry Conditions, if selected ion all occurs, and selected ion ratio is consistent with the relative abundance of standard substance, permissible variation is no more than the scope that table 1 specifies, then can contain padimate in judgement sample.
Table 1 padimate mass spectrophotometry parameter
In cosmetics of the present invention, the assay method difference from prior art of padimate is: the assay method report that there is no Padimate A and Padimate O in cosmetics at present both at home and abroad, in cosmetics of the present invention, the assay method of padimate adopts Padimate A and Padimate O in hplc simultaneous determination cosmetics, accurately, quick, highly sensitive, the quality safety Risk Monitoring for cosmetics is significant.
Below in conjunction with accompanying drawing, the assay method of padimate in cosmetics of the present invention is described further.
Accompanying drawing explanation
Fig. 1 is the ultra-violet absorption spectrum of padimate in the present invention;
Fig. 2 adopts ElipesXDBC in the present invention
18the chromatogram that (4.6mm × 250mm, 5 μm) chromatographic column obtains; Wherein, 1-Padimate A, 2-Padimate O;
Fig. 3 adopts XTerraC in the present invention
18the chromatogram that (4.6mm × 250mm, 5 μm) chromatographic column obtains; Wherein, 1-Padimate A, 2-Padimate O;
Fig. 4 adopts PromosilC in the present invention
18the chromatogram that (4.6mm × 250mm, 5 μm) chromatographic column obtains; Wherein, 1-Padimate A, 2-Padimate O;
Fig. 5 adopts SymmetryC in the present invention
18the chromatogram that (4.6mm × 250mm, 5 μm) chromatographic column obtains; Wherein, 1-Padimate A, 2-Padimate O;
Fig. 6 adopts KromasilC in the present invention
18the chromatogram that (4.6mm × 250mm, 5 μm) chromatographic column obtains; Wherein, 1-Padimate A, 2-Padimate O;
Fig. 7 is the multiple-reaction monitoring chromatogram of padimate in the present invention, wherein, and 1-Padimate A, 2-Padimate O.
The translator of Chinese of all English correspondences occurred in accompanying drawing of the present invention is:
MAU: milli absorption value; Time: time; Abs: absorbance log; Nm: nanometer; Response: response.
Embodiment
Embodiment 1
One, reagent and material
Except as otherwise noted, it is pure that agents useful for same is analysis, and water is the experimental water that GB/T6682-2008 specifies.
Methyl alcohol: chromatographically pure.
Tetrahydrofuran: chromatographically pure.
Sodium chloride.
Padimate standard items.
Padimate hybrid standard storing solution: take appropriate Padimate A and Padimate O standard items (being accurate to 0.0001g) respectively, be mixed with the list mark storing solution containing methyl alcohol 1000mg/L with methyl alcohol, then be mixed with the hybrid standard storing solution of 100mg/L with methyl alcohol.
Padimate hybrid standard working solution: with methyl alcohol padimate hybrid standard storing solution is mixed with concentration be 0.02,0.05,0.1,0.2,0.5,1,2,5,10mg/L methyl alcohol hybrid standard working solution.
Two, instrument and equipment
High performance liquid chromatograph: be furnished with UV-detector.
Analytical balance: sensibility reciprocal is 0.0001g and 0.001g.
Hydro-extractor: rotating speed is not less than 5000r/min.
Ultrasound bath
Syringe: 5mL.
Miillpore filter: 0.45 μm, organic phase.
Three, analytical procedure
1, sample preparation
(1) cream kind cosmetic sample pre-treatment
Get 1g sample and be placed in 10mL color comparison tube, after adding 2g sodium chloride breakdown of emulsion, add methyl alcohol to scale, abundant mixing, ultrasonic assistant extracts 20min, is transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, get supernatant and cross film, filtrate is for high-performance liquid chromatogram determination.
(2) lipstick class cosmetic sample pre-treatment.
Get 1g sample and be placed in 10mL color comparison tube, add 2mL tetrahydrofuran, then add methyl alcohol to scale, abundant mixing, ultrasonic assistant extracts 20min, is transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, get supernatant and cross film, filtrate is for high-performance liquid chromatogram determination.
(3) aqua class, loose powder class and the pre-treatment of shampoo class cosmetic sample
Get 1g sample and be placed in 10mL color comparison tube, add methyl alcohol to scale, fully mix, ultrasonic assistant extracts 20min, is transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, gets supernatant and cross film, and filtrate is for high-performance liquid chromatogram determination.
2, condition determination
High performance liquid chromatography reference conditions:
Chromatographic column: XTerraC
18(4.6mm × 250mm, 5 μm);
Flow velocity: 1.0mL/min;
Mobile phase: acetonitrile-water mixed solution, wherein the volume ratio of acetonitrile and water is 9:1, isocratic elution;
Determined wavelength: 310nm;
Column temperature: 30 DEG C;
Sample size: 10 μ L.
3, the drafting of typical curve
By padimate standard working solution by as above condition determination successively from low to high sample introduction measure, with peak area-concentration mapping, obtain standard regressive method, drawing standard working curve.
4, measure
By condition determination of the present invention, testing sample is measured, use quantified by external standard method.To treat in sample measuring liquid that padimate content within typical curve, should exceed after the range of linearity then should be diluted and analyzing.For adopting high performance liquid chromatography to detect positive sample, High Performance Liquid Chromatography/Mass Spectrometry/mass spectroscopy is adopted to confirm further.
5, blank test
Except not taking except sample, all undertaken by said determination condition and step.
6, result calculates
In formula:
W---the content of padimate in sample, mg/kg;
C---the concentration of padimate from the sample solution that standard working curve is found, mg/L;
V---the final constant volume of sample liquid, mL;
The quality of m---sample, g.
7, detection limit and quantitative limit
Detecting of padimate is limited to 0.2mg/kg, is quantitatively limited to 0.5mg/kg.
8, the recovery
In interpolation concentration 0.5mg/kg ~ 80000mg/kg concentration range, record the recovery 89.15% ~ 112.57%, relative standard deviation is 1.91% ~ 7.71%.
Four, the selection of experiment condition:
1, the selection of determined wavelength
Measure the maximum absorption wavelength of Padimate A and Padimate O, sweep limit is 200-400nm.After measured, as shown in Figure 1, Padimate A and Padimate O all have maximum absorption band at 310nm place.Therefore, setting its determined wavelength is 310nm.
2, the optimization of chromatographic column
Investigate ElipesXDBC respectively
18(4.6mm × 250mm, 5 μm), XTerraC
18(4.6mm × 250mm, 5 μm), PromosilC
18(4.6mm × 250mm, 5 μm), SymmetryC
18(4.6mm × 250mm, 5 μm) and KromasilC
18(4.6mm × 250mm, 5 μm) opposed polarity chromatographic column is on the impact of padimate peak shape and retention time, and result as shown in figures 2-6.Result shows, with acetonitrile: under water=90:10 is mobile phase elution requirement, various chromatographic column is hangover, peak shape and peak area indifference to padimate wash-out peak shape, and XTerraC
18(4.6mm × 250mm, 5 μm) are the most reasonable to its retention time, go out peak very fast, therefore select XTerraC
18(4.6mm × 250mm, 5 μm) are chromatographic column.
3, the optimization of mobile phase
Select methyl alcohol respectively: water=50:50, methyl alcohol: water=80:20, methyl alcohol: water=90:10, acetonitrile: water=80:20, acetonitrile: water=90:10 tetra-kinds of mobile phases, investigate the impact of different eluting solvent on padimate.Result shows, selects acetonitrile: water=90:10 is as mobile phase, and padimate elute effect peak shape is best, and retention time is the shortest, and therefore mobile phase finally selects acetonitrile: water=90:10.
4, chromatogram column temperature optimization
Investigate chromatogram column temperature goes out peak impact on padimate.Under selecting 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C conditions respectively, with acetonitrile: water=90:10 is mobile phase, XTerraC
18(4.6mm × 250mm, 5 μm) chromatographic column is analytical column, and result shows, at each temperature, and padimate retention time indifference, and under column temperature 30 DEG C of conditions, padimate peak shape is best, therefore column temperature selects 30 DEG C.
5, the selection of Extraction solvent
Relatively methyl alcohol and acetonitrile are on the impact of padimate extraction effect.Screening after measured not containing the cream kind of Padimate A and Padimate O, aqua class, loose powder class, lipstick class and shampoo class cosmetic sample as blank sample, add a certain amount of padimate standard solution wherein, after abundant mixing, extract padimate by above-mentioned steps.Result shows, using methyl alcohol and acetonitrile as sample extraction solution, to padimate extraction ratio indifference.In view of the toxicity of acetonitrile and cost higher, finally determine that methyl alcohol is sample extraction solvent.
Five, the linear relationship of method and quantitative limit
1, linear relationship
Under the determined experiment condition of this standard method, the Padimate A of preparation 1000mg/L and Padimate O standard reserving solution, prepare 100mg/L hybrid standard storing solution afterwards.Be diluted to 0.02 with methyl alcohol, 0.05,0.1,0.2,0.5,1,2,5,10mg/L; Take peak area as ordinate, corresponding padimate concentration is horizontal ordinate mapping, and in above concentration range, the linear equation of Padimate A is y=56.564x-0.1024, correlation coefficient r
2be 0.9999; The linear equation of Padimate O is y=51.417x-0.1266, correlation coefficient r
2be 0.9999, all in good linear relationship.
2, the detection limit of method and quantitative limit
Through the laboratory experiment of the method for inspection, result shows, the detection limit (S/N=3) of method is 0.2mg/kg, and quantitative limit (S/N=10) is 0.5mg/kg, and the quantitative limit high-efficient liquid phase chromatogram of Padimate A and Padimate O as shown in Figure 7.
Six, the recovery of method and precision
This standard method sets 0.5mg/kg, 5mg/kg, 10mg/kg tri-for Padimate A recovery testu and adds concentration, sets 0.5mg/kg, 500mg/kg, 80000mg/kg tri-add concentration for Padimate O recovery testu.To treat in sample measuring liquid that padimate content within typical curve, should exceed after the range of linearity then should be diluted and analyzing.By the determined experiment condition of this method of inspection, at cream kind, aqua class, lipstick class, powder class and shampoo class sample, 6 tests have all been carried out to each interpolation concentration, record the recovery 89.15 ~ 112.57%, it is 1.91 ~ 7.71% that precision test records relative standard deviation.The results are summarized in table 2-6.
The table 2 cream kind cosmetic sample recovery and Precision test result
The table 3 aqua class cosmetic sample recovery and Precision test result
The table 4 lipstick class cosmetic sample recovery and Precision test result
The table 5 loose powder class cosmetic sample recovery and Precision test result
The table 6 shampoo class cosmetic sample recovery and Precision test result
Seven, actual sample detects
The method that application is set up, to the different brands that the number of ways such as commercially available, net purchase obtain, 44 cosmetic samples of different class carry out the mensuration of padimate content, wherein 11, cream kind sample, 8, aqua class sample, 8, lipstick class sample, 8, loose powder class sample, 9, shampoo class sample.Result shows, survey in cosmetics and all do not detect Padimate A and Padimate O.
Eight, confirmatory test
For adopting high performance liquid chromatography to detect positive sample, High Performance Liquid Chromatography/Mass Spectrometry/mass spectroscopy is adopted to confirm further.
High-efficient liquid phase chromatogram condition is as follows:
Chromatographic column: HSST3 chromatographic column (100mm × 2.1mm, 1.8 μm);
Mobile phase: volume ratio is 0.1% aqueous formic acid and the methyl alcohol of 1:9;
Flow velocity: 0.4mL/min;
Column temperature: 30 DEG C;
Sample size: 10 μ L;
Mass Spectrometry Conditions is as follows:
Ion gun: electric spray ion source;
Ionization mode: positive ion mode;
Capillary voltage: 3.5kV;
Extraction voltage: 2.9V;
Ion source temperature: 150 DEG C;
Desolventizing temperature degree: 350 DEG C;
Data acquisition modes: many reactive ion monitorings.
The assay method of padimate in cosmetics of the present invention, wherein, described padimate is Padimate A and Padimate O.
The assay method of padimate in cosmetics of the present invention, wherein, described confirmatory test comprises the steps: sample liquid suitably to dilute, sample liquid and standard working solution is measured by High Performance Liquid Chromatography/Mass Spectrometry/Mass Spectrometry Conditions, if selected ion all occurs, and selected ion ratio is consistent with the relative abundance of standard substance, permissible variation is no more than the scope that table 1 specifies, then can contain padimate in judgement sample.
Table 1 padimate mass spectrophotometry parameter
Above-described embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under not departing from the present invention and designing the prerequisite of spirit; the various distortion that those of ordinary skill in the art make technical scheme of the present invention and improvement, all should fall in protection domain that claims of the present invention determines.
Claims (10)
1. the assay method of padimate in cosmetics, is characterized in that: comprise the steps:
(1) pre-treatment of sample;
(2) adopt the method for high performance liquid chromatography to analyze padimate, optimize high-efficient liquid phase chromatogram condition;
(3) drawing standard working curve, adopts quantified by external standard method, calculates the content of padimate.
2. the assay method of padimate in cosmetics according to claim 1, is characterized in that: for adopting high performance liquid chromatography to detect positive sample, adopt High Performance Liquid Chromatography/Mass Spectrometry/mass spectroscopy to confirm further.
3. the assay method of padimate in cosmetics according to claim 1, is characterized in that: described high-efficient liquid phase chromatogram condition is as follows:
Chromatographic column: XTerraC
18(4.6mm × 250mm, 5 μm);
Flow velocity: 1.0mL/min;
Mobile phase: acetonitrile-water mixed solution, wherein the volume ratio of acetonitrile and water is 9:1, isocratic elution;
Determined wavelength: 310nm;
Column temperature: 30 DEG C;
Sample size: 10 μ L.
4. the assay method of padimate in cosmetics according to claim 1, is characterized in that: described sample is cream kind, lipstick class, aqua class, loose powder class or shampoo class cosmetic sample.
5. the assay method of padimate in cosmetics according to claim 4, it is characterized in that: described sample is cream kind cosmetic sample, pre-treatment comprises the steps: that getting 1g sample is placed in 10mL color comparison tube, after adding 2g sodium chloride breakdown of emulsion, adds methyl alcohol to scale, abundant mixing, ultrasonic assistant extracts 20min, is transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, get supernatant and cross film, filtrate is for high-performance liquid chromatogram determination.
6. the assay method of padimate in cosmetics according to claim 4, it is characterized in that: described sample is lipstick class cosmetic sample, pre-treatment comprises the steps: that getting 1g sample is placed in 10mL color comparison tube, adds 2mL tetrahydrofuran, then adds methyl alcohol to scale, abundant mixing, ultrasonic assistant extracts 20min, is transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, get supernatant and cross film, filtrate is for high-performance liquid chromatogram determination.
7. the assay method of padimate in cosmetics according to claim 4, it is characterized in that: described sample is aqua class, loose powder class or shampoo class cosmetic sample, pre-treatment comprises the steps: that getting 1g sample is placed in 10mL color comparison tube, add methyl alcohol to scale, fully mix, ultrasonic assistant extracts 20min, be transferred in centrifuge tube, after the centrifugal 15min of 5000r/min rotating speed, get supernatant and cross film, filtrate is for high-performance liquid chromatogram determination.
8. the assay method of padimate in cosmetics according to claim 2, is characterized in that:
In described High Performance Liquid Chromatography/Mass Spectrometry/mass spectrum confirmation method,
High-efficient liquid phase chromatogram condition is as follows:
Chromatographic column: HSST3 chromatographic column (100mm × 2.1mm, 1.8 μm);
Mobile phase: volume ratio is 0.1% aqueous formic acid and the methyl alcohol of 1:9;
Flow velocity: 0.4mL/min;
Column temperature: 30 DEG C;
Sample size: 10 μ L;
Mass Spectrometry Conditions is as follows:
Ion gun: electric spray ion source;
Ionization mode: positive ion mode;
Capillary voltage: 3.5kV;
Extraction voltage: 2.9V;
Ion source temperature: 150 DEG C;
Desolventizing temperature degree: 350 DEG C;
Data acquisition modes: many reactive ion monitorings.
9. the assay method of padimate in cosmetics according to claim 8, is characterized in that: described padimate is Padimate A and Padimate O.
10. the assay method of padimate in cosmetics according to claim 9, it is characterized in that: described confirmatory test comprises the steps: sample liquid suitably to dilute, sample liquid and standard working solution is measured by High Performance Liquid Chromatography/Mass Spectrometry/Mass Spectrometry Conditions, if selected ion all occurs, and selected ion ratio is consistent with the relative abundance of standard substance, permissible variation is no more than the scope that table 1 specifies, then can contain padimate in judgement sample.
Table 1 padimate mass spectrophotometry parameter
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| CN105866229B (en) * | 2016-03-24 | 2018-08-07 | 中国检验检疫科学研究院 | The rapid detection method of padimate in a kind of cosmetics based on ion mobility spectrometry |
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| CN105866229B (en) * | 2016-03-24 | 2018-08-07 | 中国检验检疫科学研究院 | The rapid detection method of padimate in a kind of cosmetics based on ion mobility spectrometry |
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