CN101566603B - Method for detecting Cr(III) and Cr(5) in cosmetics by HPLC-ICP/MS combined technology - Google Patents

Method for detecting Cr(III) and Cr(5) in cosmetics by HPLC-ICP/MS combined technology Download PDF

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CN101566603B
CN101566603B CN2009100389432A CN200910038943A CN101566603B CN 101566603 B CN101566603 B CN 101566603B CN 2009100389432 A CN2009100389432 A CN 2009100389432A CN 200910038943 A CN200910038943 A CN 200910038943A CN 101566603 B CN101566603 B CN 101566603B
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刘江晖
焦红
林峰
程树军
严冬
谢湘娜
姚红
卢丽
谢玉珊
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Inspection and Quarantine Technology Center of Guangdong Entry Exit Inspection and Quarantine Bureau
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Abstract

The invention discloses a method for detecting Cr(III) and Cr(5) in cosmetics by HPLC-ICP/MS combined technology, which comprises: (1) preparing a liquid to be measured; (2) preparing a standard liquid; (3) injecting the standard liquid into a high performance liquid chromatograph for elution separation, using an inductively coupled plasma mass spectrograph to detect <52>Cr<+> , determining the retention time, the separation degree and the pulse strength of the Cr(III) and the Cr(5), taking the concentration as a horizontal coordinate and the pulse strength integral as a vertical coordinate, and obtaining the standard flexibility equation of the Cr(III) and the Cr(5) by linear fitting of an instrument computer; and (4) measuring the pulse strength integral of the Cr(III) and the Cr(5) in the liquid to be measured according to step (3), and using a standard curve to calculate and obtain the content of the Cr(III) and the Cr(5) of the liquid to be measured. The method has low detection limit and high sensitivity and accuracy.

Description

The method of Cr (III) and Cr (VI) in the HPLC-ICP/MS coupling technique detection cosmetics
Technical field
The present invention relates to chromium in a kind of cosmetics (III) and Cr (VI) content detecting method, especially related to the method that a kind of employing high performance liquid chromatography-inductivity coupled plasma mass spectrometry (HPLC-ICP/MS) coupling technique detects Cr (III) and Cr (VI) in the cosmetics.
Background technology
Chromium is argenteous hard metal, and the chromium element is a kind of element that occurring in nature extensively exists, and mainly is distributed in rock, soil, atmosphere, water and the biosome.Chromium is a kind of beidirectional element that has, and is one of trace element of needed by human, but intake also can be shown poisoning when too high, even causes death.Because chromium and compound thereof are widely used in industry such as chemical industry, plating, printing and dyeing, it is often with form contaminated air, water source and crops such as dust, steam, waste water, and invading human body by alimentary canal, respiratory tract, skin and mucous membrane influences health.
The common valence state of chromium compound has trivalent and sexavalence, and trivalent chromium and hexavalent chromium compound can transform mutually.In water body, sexavalent chrome is generally with CrO 2- 4, HCr 2O - 7, Cr 2O 2- 7Three kinds of anionic forms exist, be subjected to the influence of factors such as water pH value, temperature, redox material, organism, the toxicity of chromium exists valence state relevant with it, and trivalent chromium is one of necessary trace element of biosome, the metabolism of glucose and lipid in the control mammalian body.Sexavalent chrome has strong toxicity, is the teratogenesis carcinogen, and easily is absorbed by the body and accumulates in vivo.Chromium mainly is due to the hexavalent chromium compound to the harm of human body, solubility sexavalent chrome oxide (CrO 3 -) toxicity of the pure and mild chromium alkoxide of chromium is bigger, and have pungency and corrosivity.Zoopery proves that plumbous chromate, zinc chromate, sodium bichromate etc. have carcinogenicity.Confirm that through investigation such as epidemic diseases produce with chrome ore among the workman of dichromate, the lung cancer morbidity rate is a lot, is called " chromium lung cancer ".
Cosmetics in use directly contact the corresponding position of human body, and the Chinese government always lists cosmetics in mandatory range of management, and cosmetics quality and security are implemented strict supervision, and are particularly particularly strict to content of beary metal in the cosmetics.Regulation allows to use the chromium hydroxide green that contains trace and chrome oxide green as colorant in China's hygienic standards for cosmetics (GB7916-2007), but is not useable for oral cavity and lip cosmetic, enters in the body to prevent too much chromium.Therefore, excessive chromium is very important to the harm of human body in the cosmetics.Simultaneously, the countries concerned such as China and European Union all clearly classify the chromium element as in cosmetics banned substance.
The assay method of total chromium in the cosmetics, at present comparatively popularization and application atomic absorption spectrography (AAS) (AAS), inductively coupled plasma emission spectrography (ICP) and inductively coupled plasma mass spectrometry (ICP/MS) determination method arranged.By previous work, to China port multiple import and export brand cosmetic product such as face cream, health breast, kermes, eye shadow, kermes muffin, foundation cream, after shave powder, beautifying liquid, foundation cream dew, vanishing cream, facemask powder, skin lotion with unload cosmetics etc., measure total chromium in its cosmetics, the result shows, total chromium that all kinds of cosmetic products all exist content not wait.In fact, the detection elements total amount is to chromium element meaning and little.And the method for inspection of Cr (III) and Cr (VI) content in the cosmetics is new through looking into, and at present domestic does not still have the open industry standard method of inspection or a national standard method of inspection of approving; One, two piece of documents and materials delivering on the accidental in the world periodical, but still do not have the method for inspection that authoritative mechanism is approved.And the detection method sensitivity of general documents and materials is low, and reappearance is relatively poor, in the real work difficulty be applicable to the assay of Cr (III) and Cr (VI) in the different substrates type cosmetics, be difficult to be adapted to the demand in present international cosmetics market.
Summary of the invention
The invention provides the method that a kind of employing HPLC-ICP/MS coupling technique detects Cr (III) and Cr (VI) in the cosmetics.It is low that this method has a detectability, and sensitivity, the high superiority of accuracy are applicable to the detection of different substrates type cosmetics.
The objective of the invention is to realize by following technical measures: the method for Cr (III) and Cr (VI) in a kind of HPLC-ICP/MS coupling technique detection cosmetics, it may further comprise the steps:
(1) sample solution of preparation cosmetics to be measured: cosmetic sample is added in the edetate solution, and heating is extracted, and obtains cosmetic sample solution, moves into then in the sample introduction bottle;
(2) preparation standard solution: draw Cr (III) respectively and Cr (VI) standard stock solution dilutes, constant volume moves in the sample introduction bottle then;
(3) get standard solution and inject high performance liquid chromatograph (HPLC), adopt the reverse-phase paired ion chromatography technology to carry out the wash-out separated component, use icp ms (ICP/MS) then, adopt dynamic response pond (DRC) technology to eliminate as detecting device 40Ar 12C +With 35Cl 16OH +Right 52Cr +Spectroscopy disturb; Right 52Cr +Detect, determining Cr (III) and the retention time of Cr (VI) composition and the pulse strength of degree of separation thereof respectively, is horizontal ordinate with concentration, and the integration of pulse strength is an ordinate, make linear fit respectively by the instrument computing machine, obtain Cr (III) and Cr (VI) standard song equation;
(4) set by step (3) method is measured the pulse strength integration of Cr (III) and Cr (VI) in the cosmetic sample solution to be measured, uses the typical curve of gained to calculate the content of Cr in the cosmetic sample to be measured (III) and Cr (VI) respectively;
High performance liquid chromatograph (HPLC) condition of work in the described step (3) is: the C8 chromatographic column; Column temperature: 20～35 ℃; Mobile phase A: 0.1～2mM TBAH (TBAOH)+0.15～0.6mM edta solution; Mobile phase B: methyl alcohol (CH 3OH); Isocratic elution: mobile phase A: Mobile phase B=98～94%: 2～6%; Flow velocity 1.0～1.5mL/min; Auto injection, sample size are 20～100 μ l.
Icp ms (ICP/MS) condition of work in the described step (3): radio-frequency power (ICP RF Power): 1400～1600W; Plasma flow amount (Plasma Gas Flow) 17～20L/min; Secondary air amount (Auxiliary Gas Flow) 1.2～1.5L/min; Atomizer argon flow amount (DRC Mode NEB) 0.74～1.2L/min; Lens voltage (Lens Voltage) 6.0～10.0V; Detecting device inert stage voltage (Analog Stage Voltage)-1575～-2200V; Detector pulses step voltage (Pulse Stage Voltage) 950～1600V; Reaction gas (O 2) flow (DRC Mode CRO) 0.4～0.7mL/min; Quality pass band width (RPq) 0.4～0.7; Measure mass number (Monitored ion m/z): 52Cr +
The sample solution of described cosmetics to be measured can be prepared with method of the prior art, but for reducing the impurity peaks that when measuring, occurs, obtain stable Cr (III) and Cr (VI) composition, and obtain retention time and degree of separation preferably, described step (1) can be done following improvement: get cosmetic sample and add 5～10mL, 20～50mmoL/L pH value is in 6.0～7.2 the edetate solution, the ratio of sample quality and edetate liquor capacity is 0.3～2.0: 10～15, place the constant temperature shaking table, in 25～30 ℃ of vibration 0.5～1h, extract 2～5h at 40～60 ℃ of constant temperature of water-bath then, heated and boiled keeps 0.5～1min again; Centrifugal, get supernatant and cross 0.45 μ m film, preserve, standby.Last machine need leave standstill 20～60 minutes before measuring.
The concrete steps of described step (2) preparation standard solution: absorption 0.1～1.0mL concentration is that Cr (III) the standard stock solution of 1000 μ g/mL places beaker, add 1～10mL, 20～50mmoL/L edetate solution, heated and boiled, keep 0.5～1min, be cooled to room temperature, the nearly scale of dilute with water then, regulate PH to 6.8～7.2, adding 0.1～1.0mL concentration is Cr (VI) the standard stock solution of 1000 μ g/mL, keep PH to 6.8～7.2, shift and be settled to the 100mL volumetric flask.Get above-mentioned solution and obtain following standard serial solution with mobile phase A dilution, concentration is respectively 0.0,0.5,1.0,2.0,5.0,10.0ng/mL.Last machine need leave standstill 20～60 minutes before measuring.
Described edetate solution is edetate di-potassium solution or disodium EDTA solution.
In the analyzing and testing of trace metal ion Cr (III) and Cr (VI), very strict for the washing requirement of glassware.Glassware of the present invention-sample introduction bottle needs to soak 24 hours through salpeter solution before use, in order to avoid absorption inorganic ions in the sample introduction bottle is rinsed dry for standby then well with deionized water.Described sample introduction bottle adopts brown sample introduction bottle.Described salpeter solution is that red fuming nitric acid (RFNA) and water 1: 1 by volume are formulated.
By the recovery of standard addition test of different substrates cosmetic sample, the recovery, relative standard deviation scope meet the GB/T27404-2008 requirement, prove the feasibility of above-mentioned detection method.
The present invention compared with prior art has following remarkable result:
(1) the present invention adopts the content of Cr (III) and Cr (VI) in high performance liquid chromatography-inductivity coupled plasma mass spectrometry (HPLC-ICP/MS) coupling technique synchronous detection cosmetics, earlier by high performance liquid chromatography, adopt the composition in the reverse-phase paired ion chromatography technology wash-out sample separation solution, again by inductivity coupled plasma mass spectrometry as detecting device, adopt dynamic response pond (DRC) technology to eliminate 40Ar 12C +With 35Cl 16OH +Right 52Cr +Spectroscopy disturb.Right 52Cr +Analyze, obtain Cr (III) and the retention time of Cr (VI) composition and the pulse strength of degree of separation thereof.Concentration and pulse strength integration according to standard serial solution obtain equation of linear regression by the instrument computer fitting, calculate the content of Cr in the cosmetic sample (III) and Cr (VI) respectively.The detectability of Cr of the present invention (III) and Cr (VI) is respectively 0.05mg/kg and 0.06mg/kg, has superiority such as dynamic linear wide ranges, interference is little, analytical precision is high, analysis speed is fast.
(2) the present invention is in the sample solution of cosmetics to be measured, adopting edetate di-potassium solution or disodium EDTA solution vibration extraction then to carry out 40～60 ℃ of constant temperature after 0.5 hour extracted 2～5 hours, heated and boiled again, keep carrying out in 0.5～1 minute column front derivation, make Cr (III) and edetate di-potassium solution or disodium EDTA solution generation complex reaction in the sample, obtain Cr (III) EDTA complex compound, avoid Cr (III) in detection, to change into Cr (VI).Simultaneously, last machine left standstill 20～60 minutes before measuring, and had guaranteed the stability of Cr (III) and Cr (VI) composition, and the accuracy of content detection.
(3) the present invention fills up the technological gap of the content of Cr (III) and Cr (VI) in the domestic synchronous detection cosmetics; the new standard that poisonous element is limited the quantity of in the cosmetics is significant for working out; to protection China people's healthy and economic interests, foreign trade promotion all is of great practical significance and far-reaching historical meaning.
Description of drawings
Fig. 1 is the chromatogram that carries out the isocratic elution gained of standard solution of the present invention.
Embodiment
High performance liquid chromatography-inductivity coupled plasma mass spectrometry (HPLC-ICP/MS) coupling technique is mainly detected by high performance liquid chromatograph separation, icp ms and the interface three parts constitute.Wherein said high performance liquid chromatography adopts the reverse-phase paired ion chromatography technology to carry out the wash-out separated component; Icp ms adopts dynamic response pond (DRC) technology to eliminate spectroscopy and disturbs; It is physical interface that the coupling analysis interface of high performance liquid chromatograph and icp ms adopts quartzy eddy flow fog chamber of PC3 semiconductor refrigerating and PFA high-efficiency atomizer; Chromera morphological analysis full automatic treatment interface is a control interface.
Embodiment one: color make-up class matrix sample determination
(1) get two brown sample introduction bottles and in salpeter solution, soaked 24 hours, take out with deionized water wash then clean, drying for standby; Salpeter solution is that red fuming nitric acid (RFNA) and water 1: 1 by volume (ml/ml) are formulated.
(2) get color make-up class sample 0.3～0.5g, place 20～50mL plastics color comparison tube, add 5～10mL, 20～50mmoL/L pH value is 6.0～7.2 EDTA Dipotassium salt solution or disodium EDTA solution, put in the constant temperature shaking table, in 170～250rpm/min, 25 ℃ of vibration 0.5～1h, constant temperature 2～5 hours in 40 ℃ of water-baths then.Heated and boiled in water-bath keeps 0.5～1min again.On supercentrifuge, in 10000rpm/min, 30 ℃ of centrifugal 10～15min get supernatant and cross 0.45 μ m film then, preserve, and are standby.Last machine need leave standstill 20～60 minutes before measuring.When Cr (III) and Cr (VI) too high levels, sample solution is measured after diluting constant volume with mobile phase A.
(3) drawing 0.1～1.0mL concentration is that Cr (III) the standard stock solution of 1000 μ g/mL places beaker, adding 1～10mL 20～50mmoL/L pH value is 6.0～7.2 EDTA Dipotassium salt (or disodium salt) [EDTA] solution, heated and boiled, keep 0.5～1min, be cooled to room temperature, the nearly scale of dilute with water then, regulate PH to 6.8～7.2, adding 0.1～1.0mL concentration is Cr (VI) the standard stock solution of 1000 μ g/mL, keep PH to 6.8～7.2, transfer also is settled to the 100mL volumetric flask, obtains the solution that concentration is 1.0～10.0 μ g/mL.Get above-mentioned solution and obtain following standard serial solution with mobile phase A dilution, concentration is respectively 0.0,0.5,1.0,2.0,5.0,10.0ng/mL.Mobile phase A: the mixed solution of 0.1～2mM TBAH (TBAOH)+0.15～0.6mM EDTA Dipotassium salt (or disodium salt) [EDTA]; Mobile phase B: CH 3OH.Mobile phase A: Mobile phase B 98%: 2%.Last machine need leave standstill 20～60 minutes before measuring.
(4) getting standard solution 20～100 μ l moves in the sample introduction bottle, auto injection injects high performance liquid chromatograph (HPLC), carry out the wash-out separated component, use icp ms (ICP/MS) then, adopt dynamic response pond (DRC) technology to eliminate as detecting device 40Ar 12C +With 35Cl 16OH +Right 52Cr +Spectroscopy disturb, right 52Cr +Analyze, determine Cr (III) and the retention time of Cr (VI) composition and the pulse strength (referring to Fig. 1) of degree of separation thereof respectively, with concentration is horizontal ordinate, the integration of pulse strength is an ordinate, computing machine by instrument is made linear fit respectively, obtains Cr (III) and Cr (VI) standard song equation.
High performance liquid chromatograph (HPLC) condition of work is: the C8 chromatographic column: PerkinElmer Pecosphere Cartridge Pack C8 (3 μ mx3cm); Column temperature: 20 ℃; Mobile phase A: 0.15mM～2mM TBAH (TBAOH)+0.15～0.6mM EDTA Dipotassium salt (or disodium salt) [EDTA]; Mobile phase B: CH 3OH; Isocratic elution: mobile phase A: Mobile phase B=95%: 5%; Flow velocity 1.5mL/min.
Icp ms (ICP/MS) condition of work: radio-frequency power: 1450W; Plasma flow amount 17L/min; Secondary air amount 1.15L/min; Atomizer argon flow amount 0.74L/min; Lens voltage 9.5V; Detecting device inert stage voltage-1575V; Detector pulses step voltage 950V; Reaction gas (O 2) flow 0.55mL/min; Quality pass band width (RPq) 0.5; Measure mass number: 52Cr +
(5) sample solution with gained in the step (2) moves in the sample introduction bottle, (4) method is measured the pulse strength integration of Cr (III) and Cr (VI) in the cosmetic sample solution to be measured set by step, use above-mentioned typical curve equation respectively, automatically calculate by the instrument computing machine, can obtain the content of Cr in the cosmetic sample to be measured (III) and Cr (VI).
This method Cr (III) and Cr (VI) detect and are limited to 0.05mg/kg and 0.06mg/kg.By the recovery of standard addition test to color make-up class matrix cosmetic sample, the recovery, relative standard deviation scope meet GB/T 27404-2008 requirement, prove the feasibility of above-mentioned detection method.
The present invention can summarize with other the concrete form without prejudice to spirit of the present invention or principal character.No matter from which point, above-mentioned embodiment of the present invention all can only be thought explanation of the present invention rather than restriction, therefore in implication suitable and any change in the scope, all should think to be included in the scope of claims with claims of the present invention.

Claims (7)

1. a HPLC-ICP/MS coupling technique detects the method for Cr (III) and Cr (VI) in the cosmetics, it is characterized in that it may further comprise the steps:
(1) sample solution of preparation cosmetics to be measured: get cosmetic sample and add 5～10mL, 20～50mmoL/L pH value is in 6.0～7.2 the edetate solution, the ratio of sample quality and edetate liquor capacity is 0.3～2.0: 10～15, place the constant temperature shaking table, in 25～30 ℃ of vibration 0.5～1h, extract 2～5h at 40～60 ℃ of constant temperature of water-bath then, heated and boiled keeps 0.5～1min again; Centrifugal, get supernatant and cross 0.45 μ m film, obtain cosmetic sample solution, move into then in the sample introduction bottle;
(2) preparation standard solution: draw Cr (III) respectively and Cr (VI) standard stock solution dilutes, constant volume moves in the sample introduction bottle then;
(3) get standard solution and inject high performance liquid chromatograph, adopt the reverse-phase paired ion chromatography technology to carry out the wash-out separated component, use icp ms then, adopt the dynamic response pool technology to eliminate as detecting device 40Ar 12C +With 35Cl 16OH +Right 52Cr +Spectroscopy disturb, right 52Cr +Detect, determining Cr (III) and the retention time of Cr (VI) composition and the pulse strength of degree of separation thereof respectively, is horizontal ordinate with concentration, and the integration of pulse strength is an ordinate, make linear fit respectively by the instrument computing machine, obtain Cr (III) and Cr (VI) standard song equation;
Described icp ms condition of work: radio-frequency power 1400～1600W; Plasma flow amount 17～20L/min; Secondary air amount 1.2～1.5L/min; Atomizer argon flow amount 0.74～1.2L/min; Lens voltage 6.0～10.0V; Detecting device inert stage voltage-1575～-2200V; Detector pulses step voltage 950～1600V; Reaction gas flow speed 0.4～0.7mL/min; Quality pass band width 0.4～0.7; Measure mass number 52Cr +
(4) set by step (3) method is measured the pulse strength integration of Cr (III) and Cr (VI) in the cosmetic sample solution to be measured, uses the typical curve of gained to calculate the content of Cr in the cosmetic sample to be measured (III) and Cr (VI) respectively;
High performance liquid chromatograph condition of work in the described step (3) is: the C8 chromatographic column; Column temperature: 20～35 ℃; Mobile phase A: 0.1～2mM TBAH+0.15～0.6mM edetate solution; Mobile phase B: methyl alcohol; Isocratic elution: mobile phase A: Mobile phase B=98～94%: 2～6%; Flow velocity 1.0～1.5mL/min; Auto injection, sample size are 20～100 μ l.
2. the method for Cr (III) and Cr (VI) is characterized in that described cosmetic sample solution left standstill 20～60 minutes in the HPLC-ICP/MS coupling technique detection cosmetics according to claim 1 before last machine is measured.
3. the method for Cr (III) and Cr (VI) in the employing HPLC-ICP/MS coupling technique detection cosmetics according to claim 1 and 2, it is characterized in that, concrete steps according to described step (2) preparation standard solution: absorption 0.1～1.0mL concentration is that Cr (III) the standard stock solution of 1000 μ g/mL places beaker, add 1～10mL, 20～50mmoL/L edetate solution, heated and boiled, keep 0.5～1min, be cooled to room temperature, the nearly scale of dilute with water then, regulate PH to 6.8～7.2, adding 0.1～1.0mL concentration is Cr (VI) the standard stock solution of 1000 μ g/mL, keep PH to 6.8～7.2, shift and be settled to the 100mL volumetric flask; Get above-mentioned solution and obtain following standard serial solution with mobile phase A dilution, concentration is respectively 0.0,0.5,1.0,2.0,5.0,10.0ng/mL.
4. the method for Cr (III) and Cr (VI) is characterized in that described standard solution need leave standstill 20～60 minutes in the HPLC-ICP/MS coupling technique detection cosmetics according to claim 3 before last machine is measured.
5. the method for Cr (III) and Cr (VI) in the HPLC-ICP/MS coupling technique detection cosmetics according to claim 4, it is characterized in that described edetate solution is edetate di-potassium solution or disodium EDTA solution.
6. the method for Cr (III) and Cr (VI) in the HPLC-ICP/MS coupling technique detection cosmetics according to claim 5, it is characterized in that, sample introduction bottle in described step (1) and the step (2) soaked 24 hours through salpeter solution before use, rinse dry for standby then well with deionized water.
7. the method for Cr (III) and Cr (VI) in the HPLC-ICP/MS coupling technique detection cosmetics according to claim 6, it is characterized in that: described sample introduction bottle adopts brown sample introduction bottle.
CN2009100389432A 2009-04-24 2009-04-24 Method for detecting Cr(III) and Cr(5) in cosmetics by HPLC-ICP/MS combined technology Expired - Fee Related CN101566603B (en)

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