CN101672787B - Method for comprehensively evaluating antioxidant ability of cosmetics - Google Patents
Method for comprehensively evaluating antioxidant ability of cosmetics Download PDFInfo
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- CN101672787B CN101672787B CN2009100259617A CN200910025961A CN101672787B CN 101672787 B CN101672787 B CN 101672787B CN 2009100259617 A CN2009100259617 A CN 2009100259617A CN 200910025961 A CN200910025961 A CN 200910025961A CN 101672787 B CN101672787 B CN 101672787B
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Abstract
The invention relates to a method for comprehensively evaluating the antioxidant ability of cosmetics, comprising the following steps: preparing a series of emulsion-like base materials; dissolving cosmetics samples in the different base materials and uniformly mixed; adding detection agents for different free radicals and corresponding active oxygen/active nitrogen generation agents; respectively detecting time-dependant change values of the fluorescence intensity of the detection agents in the existence of the samples and standards and in the blank mode so as to calculate the initial reaction rate of each oxidized detection agent, obtain the antioxidant ability of the cosmetics for each free radical and further obtain the antioxidant coefficient APF value of the cosmetics. Based on maintaining the original formulation and the original status of the cosmetics, the method can evaluate the antioxidant ability of the cosmetics for each free radical, and the obtained APF value can favorably indicate the comprehensive antioxidant ability of the cosmetics. The method is applicable to multiple forms such as reagent kits, test paper, reagent sticks and the like for evaluating various skin care products and nursing products.
Description
Technical field
The present invention relates to performance evaluation, relate in particular to the integrated evaluating method of cosmetics anti-oxidant (anti-ageing) ability cosmetics such as skin care item, nursing products.
Background technology
Usually, active oxygen (ROS) and active nitrogen (NOS) comprise organism and inorganicss such as oxonium ion, free radical, superoxide, and these particles are owing to contain not paired electronics, and extremely instability has very strong activity.The main ROS/NOS that exists comprises peroxylradicals, hydroxyl radical free radical, superoxides, singlet oxygen and peroxide nitroso-in the environment.Because people's skin often is exposed in the environment, is subjected to the infringement of various free radicals easily, and skin is caused injurious effects.The scavenger of ROS/NOS (polyphenoils) can provide sharing electron, stablizes the free radical that those are in active state.By eliminating ROS/NOS, polyphenoils can effectively be defendd the active oxygen/active nitrogen in the environment, and human body skin is played a protective role.Therefore, the manufacturer begins to add in the middle of the cosmetics (as skin care item, sunblock lotion, suncream etc.) polyphenoils, with active oxygen/active nitrogen of extensively existing in the opposing environment etc.Given this, cosmetician and consumers in general wish to understand an oxidation resistance of the cosmetics of producing or using by certain technical method.Simultaneously, because cosmetics have distinctive chemical formulation and directly spread upon on the skin, they wish to understand the oxidation resistance that cosmetics are possessed when directly being applied in skin condition.
The method of traditional evaluation antioxidant ability of cosmetics as US patent 7,132,296, is that it is dissolved in certain chemical reagent, or adds solubilizer, extract etc.These methods have been destroyed the original chemical formulation of cosmetics, can not estimate it and keep original state and the oxidation resistance when directly being applied to skin.And traditional evaluation method is primarily aimed at a certain ROS/NOS and uses single detection agent, can only estimate the oxidation resistance of cosmetics at a certain free radical.Old and feeble theory is pointed out, skin senescence mainly is that various living radical long-term accumulation cause in the Skin Cell, therefore, need a series of method and estimate the oxidation resistance of cosmetics to various active oxygen/active nitrogens, the summation of each method oxidation resistance is called anti-ageing coefficient APF.According to the knowledge of the applicant, the integrated evaluating method that does not still have APF at present.
Summary of the invention
The objective of the invention is to overcome the deficiency that prior art exists, a kind of integrated evaluating method of antioxidant ability of cosmetics is provided.
For realizing purpose of the present invention, the integrated evaluating method of antioxidant ability of cosmetics, adopt following steps to carry out successively: configuration emulsion form matrix, be dissolved in the different matrix cosmetic sample and mixing, adding produces agent at the detection agent of different free radicals and corresponding active oxygen/active nitrogen, when sample exists, when standard items exist and under the space state, the fluorescence intensity that detects the central detection agent of solution respectively is worth over time, calculate the oxidized initial reaction rate of various detection agents, draw the oxidation resistance of cosmetics at various free radicals, above-mentioned several numerical value addition draws the anti-oxidant coefficient APF value of cosmetics.
Further, the integrated evaluating method of above-mentioned antioxidant ability of cosmetics, wherein, described APF value is obtained by the one or more value additions among ORAC, HORAC, NORAC, SORAC, the SOAC.
Further, the integrated evaluating method of above-mentioned antioxidant ability of cosmetics, wherein, described standard solution comprises the standard items and the matrix of detection agent, known oxidation resistance, and described blank solution comprises detection agent and matrix; Described matrix comprises oil, water and surfactant.
Further, the integrated evaluating method of above-mentioned antioxidant ability of cosmetics, wherein, described surfactant is lauryl sodium sulfate, methyl ammonium bromide or dodecyl ammonium.
Further, the integrated evaluating method of above-mentioned antioxidant ability of cosmetics, wherein, described detection agent is the DHR-6G at peroxylradicals and hydroxyl radical free radical, at the nitroso DHR-123 of peroxide, at the HE of superoxide anion and singlet oxygen.
Again further, the integrated evaluating method of above-mentioned antioxidant ability of cosmetics, wherein, described active oxygen/active nitrogen produces agent and comprises the AAPH that produces peroxylradicals, produce the CoF2/Pa/H2O2 of hydroxyl radical free radical, produce the nitroso SIN-3 of peroxidating, produce the Xanthine/Xanthine oxidase of superoxide anion, produce the Lithium Molybate/sodiumhydroxide/hydrogen peroxide of singlet oxygen.
Adopt technical solution of the present invention, can under the prerequisite that keeps original prescription of cosmetics and state, estimate the oxidation resistance of cosmetics, represent the height of its TAC by anti-oxidant (anti-ageing) coefficient APF at various free radicals.Its accuracy and comprehensive, be that prior art is incomparable, can be widely used in various skin care item and nursing product, as: sunblock lotion, suncream, cleansing cream, facial mask, shampoo, hair conditioner, Deng the comprehensive evaluation of the oxidation resistance of cosmetics, its concrete application form includes but not limited to: kit, test paper, reagent rod etc.
Description of drawings
Fig. 1 is the time dependent slope of starting stage fluorescence intensity, promptly initial reaction rate (among the figure, horizontal ordinate: the time, divide; Ordinate: fluorescence intensity; Initial Rate: initial rate);
Fig. 2 is the linear graph of the V1/V2 of standard items with its concentration change;
Fig. 3 is at peroxy radical, respectively when sample exists, when standard items exist and under the space state, the detection agent fluorescence intensity is curve over time;
Fig. 4 is at hydroxyl radical free radical, respectively when sample exists, when standard items exist and under the space state, the detection agent fluorescence intensity is curve over time;
Fig. 5 is at the peroxide nitroso-, respectively when sample exists, when standard items exist and under the space state, the detection agent fluorescence intensity is curve over time;
Fig. 6 is at superoxide anion, respectively when sample exists, when standard items exist and under the space state, the detection agent fluorescence intensity is curve over time;
Fig. 7 is at singlet oxygen, respectively when sample exists, when standard items exist and under the space state, the detection agent fluorescence intensity is curve over time.
Embodiment
Below in conjunction with specific embodiment technical solution of the present invention is described further.These examples only are some exemplary applications, can not be interpreted as a kind of restriction to claim protection domain of the present invention.
The front background portion was defended oneself, and the method for traditional detection antioxidant ability of cosmetics mainly is to add various chemical substances, solubilizer and extraction solution etc. in testing sample.Yet these methods change or have destroyed the original chemical formulation of cosmetics.Therefore, traditional detection method can not be used to detect cosmetic sample.And traditional method is only used the detection agent at a kind of active oxygen or active nitrogen sensitivity, oxidation resistance that can not the comprehensive evaluation cosmetics.
By contrast, the present invention sets up a kind of method of comprehensive evaluation antioxidant ability of cosmetics, has simultaneously kept the original prescription of cosmetics again, and has comprised main active oxygen/active nitrogen in the environment that manufacturer and consumer be concerned about.This method is used a series of detection agents at various free radicals, selects corresponding free-radical generating agent then for use.
This technical method comprises: dispose various matrix, respectively sample dissolution adds corresponding detection agent, and active oxygen/active nitrogen produces agent, detects when sample exists the time dependent situation of fluorescence intensity separately when standard items exist and under the space state.Calculate the oxidized initial reaction rate of each detection agent, draw sample anti-oxidant value at various free radicals under each detection agent.At last, gather and draw anti-ageing coefficient (APF).
This APF value is obtained by following one or more value additions: ORAC (value of anti-peroxidation free radical), HORAC (value of anti-hydroxyl radical free radical), NORAC (the nitroso value of anti-peroxide), SORAC (value of anti-superoxide anion), SOAC (value of anti-singlet oxygen).
Detection agent does not contain protein, and hydrogen atom can be provided, and comprises Dihydrorhodamine-6G (DHR-6G), Dihydrorhodamine-123 (DHR-123), hydroethidine (HE).Variety classes according to active oxygen/active nitrogen is selected the relevant detection agent: detection agent DHR-6G is at peroxylradicals and hydroxyl radical free radical; DHR-123 is at the peroxide nitroso-; HE is at superoxide anion and singlet oxygen.
Active oxygen/active nitrogen produces agent and is used for producing active oxygen/active nitrogen.2,2-Azobis (2-amidino-propane) dihydrochloride (AAPH) produces peroxylradicals, cobaltfuloride/picolinic acid/hydrogen peroxide (CoF2/Pa/H2O2) produces hydroxyl radical free radical, 3-morpholinosyndnonimine hydrochloride (SIN-3) produces the peroxidating nitroso-, Xanthine/Xanthine oxidase produces superoxide anion, and Lithium Molybate/sodiumhydroxide/hydrogen peroxide produces singlet oxygen.
Sample dissolution is in every kind of matrix, with mixer or emulsifier mixing.Each matrix all is made up of oil, water and surfactant.Standard items (comprising VE) have known oxidation resistance.Standard solution comprises the standard items and the matrix of detection agent, known oxidation resistance; Blank solution comprises detection agent and matrix.
The invention provides and estimate the serial of methods of cosmetics at the oxidation resistance of several main ROS/NOS in the environment.Here use anti-oxidant coefficient (APF) expression.Table 1 is employed several detection agents of evaluation method of the present invention and free-radical generating agent.
Table 1
| ROS/NOS | The ROS/NOS free-radical generating agent | Test item | Detection agent |
| Peroxyl Radicals (peroxy radical) | 2，2-Azobis(2-amidino-propane) dihydrochloride | ORAC | Dihydrorhodamin e-6G |
| Hydroxyl Radicals (hydroxyl radical free radical) | Cobalt?fuloride/picolinic?acid/hydrogen peroxide | HORAC | Dihydrorhodamin e-6G |
| Peroxynitrites (peroxide nitroso-) | 3-morpholinosyndnonimine?hydrochloride | NORAC | Dihydrorhodamin e-123 |
| Superoxide Anions (superoxide anion) | Xanthine/Xanthine?oxidase | SORAC | hydroethidine |
| Singlet Oxygen (singlet oxygen) | Lithium?Molybate/sodium hydroxide/hydrogen?peroxide | SOAC | hydroethidine |
The method is estimated the oxidation resistance of cosmetics, and its process may further comprise the steps:
Configuration matrix, sample solves homogeneously in the matrix by mixer or emulsifier.Matrix is made up of oil (as methyl linoleate), water and surfactant (as Tween20).This emulsion form matrix can not change or destroy the original prescription of cosmetics.Also can use other matrix that does not change or do not destroy the original prescription of cosmetics, as: oil, water and lauryl sodium sulfate; Oil, water and methyl ammonium bromide; Oil, water and dodecyl ammonium etc.
Add detection agent.Desirable detection agent does not contain protein, preferably can provide hydrogen atom.Select different detection agents at ROS/NOS different in the environment, as above shown in the table 1, also can use other non-Protein Detection agent.
Add active oxygen/active nitrogen and produce agent, be used for producing free radical at above different detection agents.When sample exists, when standard items exist and under the space state, the fluorescence intensity that detects detection agent respectively is worth over time.Standard solution comprises detection agent, standard items and matrix, and standard items have known oxidation resistance (as VE).Blank solution is made up of detection agent and matrix.
Calculate the oxidized initial reaction rate of detection agent.For example, initial rate V-120 refers to the slope that the starting stage fluorescence intensity changes among Fig. 1.Suppose that free-radical generating agent constantly produces ROS/NOS, the generating rate of ROS/NOS is constant and be assumed to be 0, and the ROS/NOS of generation and detection agent react or removed by sample or standard items.Detection agent and active oxygen/active nitrogen reaction generates the oxidation product of detection agent.Active oxygen/active nitrogen and antioxidant reaction generate the oxidation product of antioxidant.Suppose the concentration stabilize of ROS/NOS, the wear rate equation of ROS/NOS is as follows under the situation that does not have polyphenoils to exist so:
ROS/NOS wear rate equation under antioxidant exists is as follows:
Because the wear rate of ROS/NOS only produces the agent decision by it, obtain following formula:
On the other hand, when not containing antioxidant (V1) and containing antioxidant (V2), the increase formula of fluorescence intensity is respectively as equation (4) and (5):
V
1＝k
probe[probe]
1[ROS/NOS]
1…………………………………(4)
V
2＝k
probe[probe]
2[ROS/NOS]
2……………………………………(5)
Can draw from above-mentioned equation:
V
1＝k
probe[probe]
2[ROS/NOS]
2+k
antioxidant[Antioxidant]
2[ROS/NOS]
2…(6)
Therefore, rate of growth V1 and the relation of V2 of fluorescence intensity when not containing antioxidant (V1) and containing antioxidant (V2) is as follows:
V1/V2 obtains linearity curve and intercept to antioxidant concentration in (0,1), and slope k
Antioxidant/ k
Probe[probe]
2Fig. 2 provides the figure 121 of the V1/V2 of standard items with its concentration change.Slope k has been reacted the oxidation resistance of sample.Slope is big more, and oxidation resistance is strong more.The oxidation resistance accounting equation of sample is as follows:
Antioxidant?capacity＝k
Sample/k
standard………………………(8)
Fig. 3 illustrates the oxidation resistance at sample in the ORAC method of peroxy radical.Detection agent and free-radical generating agent see Table 1.When sample exists, the detection agent fluorescence intensity over time curve as 122, when standard items exist, the detection agent fluorescence intensity over time curve as 124.In this example, standard items are 370 μ M VE.Under the space state, the detection agent fluorescence intensity over time curve as 126.When sample existed, when standard items existed, under the space state, the oxidized initial reaction rate of detection agent was respectively as curve 128,130,132.With above-mentioned (1) oxidation resistance to (8) Equation for Calculating cosmetic sample.
Fig. 4 illustrates the oxidation resistance at sample in the HORAC method of hydroxyl radical free radical.Detection agent and free-radical generating agent see Table 1.When sample exists, the detection agent fluorescence intensity over time curve as 134, when standard items exist, the detection agent fluorescence intensity over time curve as 136.In this example, standard items are 18.60mM VE.Under the space state, the detection agent fluorescence intensity over time curve as 138.When sample existed, when standard items existed, under the space state, the oxidized initial reaction rate of detection agent was respectively as curve 140,142,144.With above-mentioned (1) oxidation resistance to (8) Equation for Calculating cosmetic sample.
Fig. 5 illustrates the oxidation resistance at sample in the NORAC method of peroxide nitroso-free radical.Detection agent and free-radical generating agent see Table 1.When sample exists, the detection agent fluorescence intensity over time curve as 146, when standard items exist, the detection agent fluorescence intensity over time curve as 148.In this example, standard items are 464 μ M VE.Under the space state, the detection agent fluorescence intensity over time curve as 150.When sample existed, when standard items existed, under the space state, the oxidized initial reaction rate of detection agent was respectively as curve 152,154,156.With above-mentioned (1) oxidation resistance to (8) Equation for Calculating cosmetic sample.
Fig. 6 illustrates the oxidation resistance at sample in the SORAC method of superoxide anion.Detection agent and free-radical generating agent see Table 1.When sample exists, the detection agent fluorescence intensity over time curve as 158, when standard items exist, the detection agent fluorescence intensity over time curve as 160.In this example, standard items are 2 μ M VE.Under the space state, the detection agent fluorescence intensity over time curve as 162.When sample existed, when standard items existed, under the space state, the oxidized initial reaction rate of detection agent was respectively as curve 164,166,168.With above-mentioned (1) oxidation resistance to (8) Equation for Calculating cosmetic sample.
Fig. 7 illustrates the oxidation resistance at sample in the SOAC method of singlet oxygen.Detection agent and free-radical generating agent see Table 1.When sample exists, the detection agent fluorescence intensity over time curve as 170, when standard items exist, the detection agent fluorescence intensity over time curve as 172.In this example, standard items are 18.60 μ M VE.Under the space state, the detection agent fluorescence intensity over time curve as 174.When sample existed, when standard items existed, under the space state, the oxidized initial reaction rate of detection agent was respectively as curve 176,178,180.With above-mentioned (1) oxidation resistance to (8) Equation for Calculating cosmetic sample.
The present invention estimates the method for antioxidant ability of cosmetics need not add other chemical substance, as solubilizer, and extract etc.Therefore, this method does not change or destroys original prescription of cosmetics, and provides product to be used in the evaluation method of oxidation resistance under the skin virgin state.In addition, this method can be estimated the oxidation resistance to several main free radicals in the environment.
At several main free radicals in the environment, the present invention can provide comparatively comprehensively oxidation resistance integrated evaluating method, draws the anti-oxidant coefficient (APF) of cosmetics.Specifically comprise: dispose a series of matrix, with sample dissolution mixing in different matrix, adding is at the detection agent of different free radicals and at the free-radical generating agent (seeing Table 1) of different detection agents, when test sample exists respectively, when standard items exist and under the space state, each detection agent fluorescence intensity over time, calculate the initial reaction rate (square journey 1-8) of every kind of detection agent, draw the oxidation resistance of cosmetics at every kind of free radical, several value additions obtain the APF value.
One instantiation according to the present invention, the APF value of certain cosmetic sample is obtained by the anti-oxidant value addition of following several method: ORAC, HORAC, NORAC, SORAC, SOAC.
Certainly, the APF value be not limited to above-mentioned several method value and, also comprise other similar anti-oxidant value.In sum, can draw to draw a conclusion: the present invention can be at the oxidation resistance of several free radical comprehensive evaluation cosmetics that exist in the environment, and draws total antioxidation coefficient (APF value).This method evaluation result is accurate, objective, can be widely used in all kinds of skin care nursing materialss, and its concrete application form includes but not limited to: kit, test paper, reagent rod etc.
Claims (1)
1. the integrated evaluating method of antioxidant ability of cosmetics, it is characterized in that: one or more in peroxy radical ORAC, the hydroxyl radical free radical HORAC of cosmetics, peroxide nitroso-NORAC, superoxide anion SORAC, these five kinds of free radicals of singlet oxygen SOAC, detect according to the following steps and estimate---
Configuration emulsion form matrix, be dissolved in the matrix cosmetic sample and mixing, adding produces agent at the detection agent of different free radicals and corresponding active oxygen/active nitrogen, when sample exists, when standard items exist and under the space state, the fluorescence intensity that detects the central detection agent of solution respectively is worth over time, calculate the oxidized initial reaction rate of various detection agents, draw the oxidation resistance of cosmetics at various free radicals, wherein both or both above addition obtains the anti-oxidant coefficient APF value of these cosmetics;
In the testing process, described matrix comprises oil, water and surfactant, and described surfactant is lauryl sodium sulfate, methyl ammonium bromide or dodecyl ammonium;
Described detection agent is the DHR-6G dihydro rhodamine 6G at peroxylradicals and hydroxyl radical free radical, at the nitroso DHR-123 dihydro of peroxide rhodamine 123, at the HE dihydro second ingot fluorescent dye of superoxide anion and singlet oxygen;
Described active oxygen/active nitrogen produces agent and comprises: produce 2 of peroxylradicals, 2 '-azo diisobutyl amidine dihydrochloride, produce cohalt difluoride, pyridine carboxylic acid and the hydrogen peroxide of hydroxyl radical free radical, produce the nitroso 5-amino-3-of peroxidating (4-morpholinyl)-1,2,3-oxadiazoles hydrochloride, the xanthine and the xanthine oxidase of generation superoxide anion, lithium molybdate, NaOH and the hydrogen peroxide of generation singlet oxygen;
Described standard solution comprises the standard items and the matrix of detection agent, known oxidation resistance, and described blank solution comprises detection agent and matrix.
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| CN107315000A (en) * | 2017-06-22 | 2017-11-03 | 湖北工业大学 | A kind of method for analyzing oil-soluble inhibitor oxidation resistance in emulsion |
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| CN107271606B (en) * | 2013-07-19 | 2020-07-03 | 荷兰联合利华有限公司 | Method of evaluating the efficacy of a personal care product |
| CN105675464A (en) * | 2016-01-27 | 2016-06-15 | 曼秀雷敦(中国)药业有限公司 | Method for evaluating anti-pollution effect of washing type cosmetic |
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| US7132296B2 (en) * | 2002-02-15 | 2006-11-07 | Medical Products Manufacturing, Llc | Method for assaying the antioxidant capacity of a sample |
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| US7132296B2 (en) * | 2002-02-15 | 2006-11-07 | Medical Products Manufacturing, Llc | Method for assaying the antioxidant capacity of a sample |
| CN101377471A (en) * | 2007-08-27 | 2009-03-04 | 英科新创(厦门)科技有限公司 | Electrochemical method for evaluating antioxidant ability of substance to be tested |
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| CN107315000A (en) * | 2017-06-22 | 2017-11-03 | 湖北工业大学 | A kind of method for analyzing oil-soluble inhibitor oxidation resistance in emulsion |
| CN107315000B (en) * | 2017-06-22 | 2019-09-17 | 湖北工业大学 | A method of analysis oil-soluble inhibitor oxidation resistance in lotion |
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