CN108375636A - The detection method of Arsenic in Cosmetics compound - Google Patents
The detection method of Arsenic in Cosmetics compound Download PDFInfo
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- CN108375636A CN108375636A CN201810103337.3A CN201810103337A CN108375636A CN 108375636 A CN108375636 A CN 108375636A CN 201810103337 A CN201810103337 A CN 201810103337A CN 108375636 A CN108375636 A CN 108375636A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
Abstract
The present invention provides a kind of detection method of Arsenic in Cosmetics compound, includes the following steps：(1) detected sample is taken, according to 1g:The ratio of 10~20ml is uniformly mixed with 2mol/L phosphate aqueous solutions；(2) mixture is in 60~70 DEG C of 8~12min of microwave abstracting；(3) after the product cooling of step (2), with 8000r/min rotating speeds in 4 DEG C of centrifugal treating 10min, supernatant crosses 0.45 μm of organic filter membrane, obtains sample liquid；(4) with arsenic content in high performance liquid chromatography and inductivity coupled plasma mass spectrometry combination detection sample liquid.The method of the present invention convenient can delicately detect the arsenic compound of micro existing variform in cosmetics, including inorganic arsenic As III and As V and methanearsonic acid MMA and cacodylic acid DMA.
Description
Technical field
The invention belongs to daily necessities field of quality control, it particularly relates to toxic residua in cosmetics and its raw material
Detection method.
Background technology
Before people can trace back to several centuries to the understanding of arsenic, colourless, tasteless arsenic trioxide powder (arsenic)
It is often used as hypertoxic drug.Arsenic is all classified as the mankind by international cancer research institution (IARC) and Environmental Protection Agency USA (EPA)
Carcinogenic substance.The arsenic of different shape has different physics and chemical property, and toxicity is also different, wherein inorganic arsenic (III Hes of As
As V) toxicity it is maximum, the toxicity of the organo-arsenics such as methanearsonic acid (MMA) and cacodylic acid (DMA) is smaller, and arsenic glycine betaine
(AsB), arsenocholine (AsC) and arsenic sugar are typically considered nontoxic.The existing forms of arsenic are increasingly concerned by people, and are distinguished
Organo-arsenic in product and inorganic arsenic, it is particularly important for safety evaluatio.The morphological analysis of arsenic mainly use liquid chromatogram with
The joint technology such as inductivity coupled plasma mass spectrometry (ICP-MS), atomic fluorescence spectrophotometry (AFS), atomic absorption spectrum (AAS).By
Have many advantages, such as that high sensitivity, detection limit are low, the range of linearity is wide in ICP-MS, HPLC-ICP-MS becomes morphological analysis most
One of method for combined use of application prospect is widely used for the Senile Mouse of Food and environment sample.And Arsenic in Cosmetics
Form is more, and separation analysis difficulty is very big, has not been reported.
The extracting method of inorganic arsenic mostly uses hydrochloric acid at present or salpeter solution water-bath is impregnated, and method extraction time is longer, pole
It easily leads to other arsenic morphologies in sample to be extracted into extracting solution, keeps measurement result higher, and salpeter solution extraction easily occurs
Valence state converts between As III and As V.Accurate measurements needs are obtained in arsenium morphological analysis to keep as possible in sample handling processes
It is very crucial the processing procedures such as to detach with sample, purify for the chemical constitution of its original concentration and original form, correctly sampling.
Invention content
The purpose of the present invention is to provide a kind of methods can be used for a variety of arsenic compound analyses in cosmetics, especially needle
To the great inorganic arsenic chemicals of wherein harmfulness.
The detection method of the Arsenic in Cosmetics compound, includes the following steps：
(1) detected sample is taken, according to 1g:The ratio of 10~20ml is uniformly mixed with 2mol/L phosphate aqueous solutions；
(2) mixture is in 60~70 DEG C of 8~12min of microwave abstracting；
(3) after the product cooling of step (2), with 8000r/min rotating speeds in 4 DEG C of centrifugal treating 10min, supernatant crosses 0.45
μm organic filter membrane, obtains sample liquid；
(4) in high performance liquid chromatography (HPLC) and inductivity coupled plasma mass spectrometry (ICP-MS) combination detection sample liquid
Arsenic content.
The present invention establishes the above-mentioned detection method for Arsenic in Cosmetics, convenient can delicately detect micro- in cosmetics
The arsenic compound of the existing variform of amount, including inorganic arsenic As III and As V and methanearsonic acid MMA and cacodylic acid
DMA.Under the detection architecture that this method is established, the detection limit in terms of arsenic of trivalent arsenic As III and pentavalent arsenic As V are respectively
5.0 and 7.0 μ g/kg, the range of linearity are 5~100 μ g/L, and related coefficient is all higher than 0.9995, recovery of standard addition is 82.1%~
Between 103.8%, relative standard deviation 3.23%~5.10%, n=6.
Description of the drawings
3 width of attached drawing of the present invention：
Fig. 1 is influence test result of the EDTA concentration to arsenic retention time.
Fig. 2 is influence test results of the pH to arsenic retention time.
Fig. 3 is different shape arsenic standard solution HPLC-ICP-MS spectrograms
Specific implementation mode
The present invention provides a kind of detection method of Arsenic in Cosmetics compound, includes the following steps：
(1) detected sample is taken, according to 1g:The ratio of 10~20ml is uniformly mixed with 2mol/L phosphate aqueous solutions；
(2) mixture is in 60~70 DEG C of 8~12min of microwave abstracting；
(3) after the product cooling of step (2), with 8000r/min rotating speeds in 4 DEG C of centrifugal treating 10min, supernatant crosses 0.45
μm organic filter membrane, obtains sample liquid；
(4) in high performance liquid chromatography (HPLC) and inductivity coupled plasma mass spectrometry (ICP-MS) combination detection sample liquid
Arsenic content.
In specific implementation mode, the microwave abstracting in the step (2) preferably uses polytetrafluoroethylcontainer container, in 65 DEG C of extractions
Take 10min.
In another embodiment, more efficiently to distinguish and measuring a variety of arsenic compounds in sample, the step (4)
Middle high performance liquid chromatography (HPLC) most preferably uses：Anion guard column Hamilton PRP-X100,25mm × 4.1 mm；It is cloudy
Ion analysis column Hamilton PRP-X100,250mm × 4.1mm；Mobile phase is containing 0.2mmol/L EDTA and 2mmol/L
NaH2PO4Aqueous solution；Flowing phase pH value is 6.0；Flow velocity：1.0mL/min；Sample size：50μL.It is connected using PEEK pipelines
ICP-MS and chromatographic column.Its mobile phase preferably comprises the methanol of 1% (volume fraction).
To realize, accurately detection, most preferred testing conditions include：Inductivity coupled plasma mass spectrometry in the step (4)
(ICP-MS) parameter is：Radio-frequency power：1300W；Reflection power：2W；Carrier gas：High-purity helium 0.85L/min；Assist gas：It is high-purity
Helium 0.12L/min；Sampling depth：6.5mm；It is atomized room temperature：2℃；Atomizer：Concentric spray chamber and standard quartz torch.
With reference to non-limiting embodiment, the present invention will be further described, is not construed as to arbitrary shape of the present invention
The restriction of formula.In these embodiments, used HPLC-ICP-MS detection devices and condition, according to cited hereinabove
Most preferred embodiment carries out.
In this specification, unless otherwise specified, involved instrument includes in this specification：7500ce type inductive couplings etc.
Gas ions mass spectrograph (Agilent companies of the U.S.)；1100 type high performance liquid chromatography of HPLC (Agilent companies of the U.S.)；MLS-
1200 type microwave digestion devices (Italian Milestone companies)；Milli-Q ultrapure water systems (French Millipore Corp.)；It is high
Fast refrigerated centrifuge (German Hermle companies).
Described in this specification and sodium dihydrogen phosphate, disodium ethylene diamine tetraacetate, absolute ethyl alcohol, sodium hydroxide, phosphoric acid,
Nitric acid, hydrogen peroxide are top pure grade；Pure water (18.2M Ω cm)；Mass spectrum tunes liquid：7Li、89Y、140Ce、205Tl, mass concentration
For 10 μ g/L, it is purchased from Agilent companies.Arsenate (As III), arsenous anion (As V), methanearsonic acid (MMA) and dimethyl arsine
Sour (DMA) is purchased from State Standard Matter Research Centre.All storing solutions are kept in dark place in 4 DEG C of refrigerations, mixed standard solution by
Standard reserving solution dilutes preparation with pure water step by step.
Embodiment 1. extracts influence of the solution to arsenic recovery rate
This experiment, for detected sample, is distinguished with the commercially available toner containing inorganic arsenic, facial cleanser, shampoo, muffin etc.
2 parallel testing groups are set.Each test group takes 1~2 gram of detected sample respectively, according to 1g:The ratio of 20ml from it is different
Extraction solvent is uniformly mixed；65 DEG C of microwave abstracting 10min；After product cooling, with 8000r/min rotating speeds in 4 DEG C of centrifugal treatings
10min, supernatant cross 0.45 μm of organic filter membrane, obtain sample liquid；Then the recovery rate of arsenic is detected and calculated with HPLC-ICP-MS.
Wherein, the other Extraction solvent of each group is respectively：
A. mass concentration is 50% methanol-water solution；
B. pure water；
C. the hydrochloric acid solution of a concentration of 1mol/L, 2mol/L, 3mol/L, 5mol/L；
D. the salpeter solution of a concentration of 1mol/L, 2mol/L, 3mol/L, 5mol/L；
E. the acetic acid solution of a concentration of 1mol/L, 2mol/L, 3mol/L, 5mol/L；
F. a concentration of 1mol/L, 2mol/L, 3mol/L, 5mol/L phosphate aqueous solution；
The computational methods of recovery rate are：Sample to be tested extracts according to the Extraction solvent of a~f and measures nothing according to the method described above
Machine arsenic content, then according to 1g:The ratio of 20ml is extracted with 2mol/L phosphate aqueous solutions and measures inorganic arsenic after mixing to be contained
Amount, the two ratio is recovery rate.
50% methanol solution, pure water, hydrochloric acid solution, salpeter solution, acetic acid solution, phosphoric acid solution etc. have been investigated in experiment respectively
Different extraction systems.The result shows that a kind of any of the above described extractant, the extraction efficiency of inorganic arsenic can reach 50% or more,
The extraction efficiency of wherein pure water is relatively low, is especially unsuitable for the cosmetic samples such as paste, powder.When 50% methanol solution extracts
Efficiency is higher, but pre-treatment, which needs nitrogen to blow or heat, removes methanol, operates loss that is relatively complicated and may causing inorganic arsenic.
Hydrochloric acid solution introduces ICP-MS, and chlorine argon interferes the measurement that can influence arsenic.The extraction efficiency of 5mol/L salpeter solutions is some higher, but
It is since oxidisability is strong, part As (III) is oxidized to As (V) in sample in extraction sample, and the form of arsenic is converted.
The extraction efficiency of acetic acid and phosphoric acid relatively, imitates arsenic generation matrix enhanced sensitivity it is contemplated that the introducing of carbon is easy in acetic acid
It answers, selects to determine 2mol/L phosphoric acid solutions as extracting solution eventually by optimization concentration.
Influence of 2. Extracting temperature of embodiment to arsenic recovery rate
This experiment, for detected sample, is distinguished with the commercially available toner containing inorganic arsenic, facial cleanser, shampoo, muffin etc.
2 parallel testing groups are set.Each test group takes 1~2 gram of detected sample respectively, according to 1g:The ratio and 2mol/L of 20ml
Phosphoric acid solution is uniformly mixed；It is extracted according to condition of different temperatures；After product cooling, with 8000r/min rotating speeds in 4 DEG C of centrifugations
10min is handled, supernatant crosses 0.45 μm of organic filter membrane, obtains sample liquid；Then it is detected with HPLC-ICP-MS and calculates carrying for arsenic
Take rate.
Wherein, the other extraction conditions of each group are respectively：
A. room temperature extraction 2h, 4h, 8h, 12h, for 24 hours；
DEG C b.50 extraction 2h, 4h, 8h, 12h, 16h, 18h, for 24 hours；
DEG C c.60 extraction 2h, 4h, 8h, 12h, 16h, 18h, for 24 hours；
DEG C d.70 extraction 2h, 4h, 8h, 12h, for 24 hours；
DEG C e.80 extraction 2h, 4h, 8h, 12h, for 24 hours；
F.60 DEG C microwave abstracting 5min, 10min, 15min；
G.65 DEG C microwave abstracting 5min, 10min, 15min；
H.70 DEG C microwave abstracting 5min, 10min, 15min；
The computational methods of recovery rate are：Sample to be tested extracts according to the temperature condition of a~h and measures nothing according to the method described above
Machine arsenic content, then extracts and measures abio-arsenic residues according to 65 DEG C of microwave abstracting 10min, and the two ratio is recovery rate.
Extracting temperature is also an important factor for determining inorganic arsenic extraction efficiency, when investigating phosphoric acid solution as extracting solution herein
Different bath temperature conditions.The result shows that when temperature is 60 DEG C, the extraction efficiency of extraction 16~18h inorganic arsenics is needed to reach
To highest；When temperature reaches 80 DEG C or more, hot dipping carries 2~4h of needs, is substantially shorter by the way of Microwave-assisted firing
Extraction time, 65 DEG C of microwave abstracting 10min can reach satisfied extraction efficiency.
For the white class cosmetic sample in part, containing certain emulsifier component, these emulsifiers hold in extraction process
Solution is easily set to form emulsion, so as to cause layered filtration is difficult to, common demulsification mode has addition sodium chloride, heating or in refrigerator
Middle standing, wherein the mode of addition sodium chloride can cause salt content in sample solution excessively high, and detection of the chloride to arsenic
It may cause a degree of interference.Sample is handled herein by the mode of microwave heating, it is found that most of sample is equal
It can meet the requirements, the sample of incomplete demulsification after heating is centrifuged for 4 DEG C with 8000r/min rotating speeds low temperature, can also be played certain
Effect.
The optimization of 3. mobile phase of embodiment
With the commercially available facial cleanser containing inorganic arsenic, shampoo etc., for detected sample, 2 parallel surveys are respectively set in this experiment
Examination group.Each test group takes 1~2 gram of detected sample respectively, according to 1g:The ratio of 20ml mixes with 2mol/L phosphoric acid solutions
It is even；65 DEG C of microwave abstracting 10min；After product cooling, with 8000r/min rotating speeds in 4 DEG C of centrifugal treating 10min, supernatant mistake
0.45 μm of organic filter membrane, obtains sample liquid；Then different mobile phase solvents, HPLC-ICP-MS is used to detect the inorganic arsenic in sample.
Wherein, the other mobile phase solvent of each group is respectively：
a.0mol/L NaH2PO4；
b.2mol/L NaH2PO4；
c.4mol/L NaH2PO4；
d.6mol/L NaH2PO4；
e.8mol/L NaH2PO4；
f.2mol/L NaH2PO4+0.5mmol/L EDTA；
g.2mol/L NaH2PO4+1.0mmol/L EDTA；
h.2mol/L NaH2PO4+2.0mmol/L EDTA；
i.2mol/L NaH2PO4+3.0mmol/L EDTA；
Arsenic morphology separation at present mostly uses ion exchange mechanism, has selected anion-exchange column to be detached herein, phosphoric acid
The isocratic elution of salting liquid.NaH in mobile phase is investigated2PO4A concentration of 0~8mmol/L when chromatographic peak separation situation, as a result
Show NaH2PO4Concentration the chromatographic isolation of inorganic arsenic is influenced it is smaller, to avoid the shadow of excessively high sodium salt plasma square
It rings, NaH2PO4A concentration of 2mmol/L.
EDTA is added in mobile phase can eliminate loss of the arsenic in chromatographic separation process, investigate various concentration EDTA
Influence to arsenic appearance time, the results are shown in Figure 1.As can be seen that the increase of EDTA concentration can shorten the guarantor of DMA and As III
The time is stayed, when 0.2mmol/LEDTA is added in mobile phase, separating effect is best.
The optimization of 4. mobile phase pH of embodiment
With the commercially available facial cleanser containing inorganic arsenic, shampoo etc., for detected sample, 2 parallel surveys are respectively set in this experiment
Examination group.Each test group takes 1~2 gram of detected sample respectively, according to 1g:The ratio of 20ml mixes with 2mol/L phosphoric acid solutions
It is even；65 DEG C of microwave abstracting 10min；After product cooling, with 8000r/min rotating speeds in 4 DEG C of centrifugal treating 10min, supernatant mistake
0.45 μm of organic filter membrane, obtains sample liquid；Using 2mmol/LNaH2PO4With 0.2mmol/LEDTA as mobile phase, different pH items
Part detaches, and HPLC-ICP-MS detects the inorganic arsenic in sample.
Wherein, the other mobile phase pH of each group is respectively：
a.4.0；
b.6.0；
c.8.0；
d.10.0；
With 2mmol/LNaH2PO4With 0.2mmol/LEDTA as mobile phase, pH colors under the conditions of 4.0~10.0 are had studied
The separation situation of spectrum.The results are shown in Figure 2, it is seen that：PH is to MMA, the obvious effect of AsV retention times, and to As (III)
Influence is smaller, is primarily due to the pKa=9.2 of As III, As III is with H before pH 9.23AsO3Form exist, not with ion
It has an effect to reagent, in chromatographic column almost without reserve.The retention time of DMA is in pH>When 8.0, contract with the increase of pH
It is short.The retention time of MMA shortens between pH 4.0~8.0 as pH increases, and whens 6.0~8.0 pH is held essentially constant,
Increase again as pH increases retention time after pH 8.0.In pH 4.0~6.0, this range is gradually shortened AsV retention times,
This range of pH 6.0~8.0 increases, this range of pH 8.0~10.0 shortens again.In view of shortening analysis time, mobile phase
It is 6.0 to be set as pH value, while adjusting leacheate pH value using phosphoric acid and sodium hydroxide to avoid Matrix effects.
In addition, the addition of methanol can effectively improve sensitivity and shorten appearance time, it is contemplated that organic compound combustion
Influence to ICP-MS taper holes determines that methanol content is 1% (V/V) in leacheate.The results are shown in Figure 3, it is seen that：Standard solution
In 4 kinds of arsenic morphologies have been carried out and be kept completely separate in 12min.
Embodiment 5.
This experiment to a concentration of 5.0,10.0,20.0,50.0,100.0ug/L V mixed standard solution of As III and As, with
PH=6.0,2mmol/LNaH2PO4The methanol aqueous solution that volume fraction with 0.2mmol/LEDTA is 1% is mobile phase, is carried out
HPLC-ICP-MS is analyzed, and linear regression is carried out to concentration with the peak area of each arsenic morphology.
The result shows that the linear regression coeffficient of As III and As V are respectively 0.9996,0.9995.According to minimum concentrations
And the extension rate of sample treatment, the method detection that inorganic arsenic in cosmetics is calculated are limited to：III 5.0ug/ of trivalent arsenic As
V 7.0ug/kg of kg, pentavalent arsenic As.
6. precision of embodiment and mark-on reclaims measure
With the commercially available toner containing inorganic arsenic, facial cleanser, shampoo etc., for detected sample, 3 surveys are arranged in this experiment
Examination group.Each test group takes 1~2 gram of detected sample respectively, quantitatively adds V standard of As III and As of different concentration levels
Solution so that As III and V each mark-on amounts of As are 0.05,0.10 and 0.50 μ g, according to 1g:The ratio of 20ml and 2mol/L phosphoric acid
Solution is uniformly mixed；65 DEG C of microwave abstracting 10min；After product cooling, with 8000r/min rotating speeds in 4 DEG C of centrifugal treating 10min,
Supernatant crosses 0.45 μm of organic filter membrane, obtains sample liquid；HPLC-ICP-MS detects the inorganic arsenic in sample.
Concrete outcome is shown in Table 1, and by calculating, the rate of recovery of each mark-on level is opposite to mark between 82.1%~103.8%
Quasi- deviation 3.23%~5.10%.
The measurement result of the precision and mark-on reclaims of 1 method of table
The application of 7. actual sample of embodiment
Chinese Cosmetic Market is broadly divided into skin care, beauty, hairdressing and other types cosmetics.Choose commercially available wash one's face
Milk, toner, nutrient solution, face cream, shampoo, hair care breast, gel, blush, cake cosmetics sample, accurately weigh more parts it is parallel
Sample, ICP-MS directly measures the content of total arsenic after resolution, while the content of inorganic arsenic is measured with the method established, and as a result arranges
In table 2.The results show that the pollution condition difference for the Arsenic in Cosmetics that experiment is chosen is larger, but it is less than national health mark
Quasi- limitation, the form of arsenic mostly exist with inorganic arsenic, especially trivalent arsenic, and the content of beauty class product arsenic poor quality is higher,
Inorganic arsenic and organo-arsenic are detected in which part skin-protection product and muffin, blush class product simultaneously.
Measurement result (the unit of 2 sample of table：Mg/kg, n=3)
In table 2, N.D is to be not detected.
Claims (5)
1. the detection method of Arsenic in Cosmetics compound, includes the following steps：
(1) detected sample is taken, according to 1g:The ratio of 10~20ml is uniformly mixed with 2mol/L phosphate aqueous solutions；
(2) mixture is in 60~70 DEG C of 8~12min of microwave abstracting；
(3) after the product cooling of step (2), with 8000r/min rotating speeds in 4 DEG C of centrifugal treating 10min, supernatant, which crosses 0.45 μm, to be had
Machine filter film, obtains sample liquid；
(4) with arsenic content in high performance liquid chromatography and inductivity coupled plasma mass spectrometry combination detection sample liquid.
2. detection method according to claim 1, which is characterized in that the microwave abstracting operating parameter in the step (2)
For：Polytetrafluoroethylcontainer container, 65 DEG C of extraction 10min.
3. detection method according to claim 1, which is characterized in that high performance liquid chromatography (HPLC) in the step (4)
It is：Anion guard column Hamilton PRP-X100,25mm × 4.1mm；Anion analysis column Hamilton PRP-X100,
250mm×4.1mm；
Mobile phase is containing 0.2mmol/L EDTA and 2mmol/L NaH2PO4Aqueous solution；
Flowing phase pH value is 6.0；
Flow velocity：1.0mL/min；Sample size：50μL.
4. detection method according to claim 3, which is characterized in that also contain according to percent by volume in the mobile phase
There is 1% methanol.
5. detection method according to claim 1, which is characterized in that inductively coupled plasma matter in the step (4)
Composing (ICP-MS) parameter is：Radio-frequency power：1300W；Reflection power：2W；Carrier gas：High-purity helium 0.85L/min；Assist gas：It is high
Pure helium 0.12L/min；Sampling depth：6.5mm；It is atomized room temperature：2℃；Atomizer：Concentric spray chamber and standard quartz torch.
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Application publication date: 20180807 |