CN109115761A - The detection method of tranexamic acid in cosmetics - Google Patents
The detection method of tranexamic acid in cosmetics Download PDFInfo
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- CN109115761A CN109115761A CN201811361462.0A CN201811361462A CN109115761A CN 109115761 A CN109115761 A CN 109115761A CN 201811361462 A CN201811361462 A CN 201811361462A CN 109115761 A CN109115761 A CN 109115761A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
Abstract
The present invention provides a kind of detection method of tranexamic acid in cosmetics, specifically comprise the following steps: that (A) successively adds reagent A in sample to be tested, reagent B shakes up, isolated supernatant；(B) after addition reagent C shakes up layering in supernatant, separation subnatant adds color developing agent, and the situation that develops the color is observed in 90 DEG C of heating；Wherein, the reagent A is at least one of Tween-20, Tween-80, PEG-20000, and the reagent B is DMSO, DMF, EtOH, CH2Cl2At least one of, the reagent C is at least one of water, saturated sodium chloride solution, saturation phosphoric acid hydrogen dimethylamino solution, saturation phosphoric acid the second light industry bureau potassium solution, saturation disodium phosphate soln, saturation sodium dihydrogen phosphate.Detection method of the invention is easy to operate, high-efficient, at low cost, should be widely promoted and is applied.
Description
Technical field
The present invention relates to cosmetics qualities to detect control field, in particular to tranexamic acid in a kind of cosmetics
Detection method.
Background technique
Tranexamic acid (also known as Trenaxmine) is a kind of protease inhibitors, can catalysis of the protease inhibition to peptide bond hydrolysis
Effect, to prevent the activity of such as inflammatory protease enzyme, and then inhibit blackspot position epidermal cell function it is mixed
Disorderly, and inhibit melanin enhancement factor group, then thoroughly break off the approach occurred because of the melanin that ultraviolet light irradiates and is formed.
Blackspot is allowed no longer to thicken, expand and increase, so as to be effectively prevented and improve the pigment deposition of skin.
Medically it is applied to the bleed bottom diagnosis and treatment of aneurysm spider nethike embrane, educational circles is referred to as disconnected blood inflammation and uses as coagulant
On the way, diagnosis and treatment process has been surprisingly found that the side effect for making skin bleach, so medicinal with melanin Shen Dian using treating in chloasma hepaticum
Medical prescription pad.The whitening mechanism of Trenaxmine be while and rapidly inhibit Tyrosine Enzyme ferment and melanocyte activity, and
The case where preventing melanin from assembling, can block prevent because ultraviolet light irradiate and formed melanin deterioration travel path.
Trenaxmine can give expression to effect stability compared with traditional whitening composition, be limited by high temperature production environment, not need each
The protection transmission effect of class carrier, can still reach whitening effect, have no irritation effect.Its external application can inhibit melanin to be formed
And the formation of pigmented spots is prevented, the path of melanin transmitting is blocked, Department of Health's normal concentration limitation is 2%-3%.In whitening
Cheng Zhong, based on Trenaxmine, whitening raw materials are done in collocation vitamin C magnesium phosphate (APMg).
There are many whitening spot-removing class cosmetics on the market now the ingredient containing tranexamic acid is directly added into cosmetics,
But due to being the usage except legal indication, drug safety is not can guarantee, once cause legal dispute, processing can be very complicated.
Tranexamic acid is that clinically common hemostatic, abuse potential cause the central nervous systems such as eye-blurred, headache, dizziness, tired
System symptom；Intracranial thrombosis and bleeding excessively can be caused, causes secondary renal plevis inflammation and ureter sludged blood to be blocked, additionally
There are the symptoms such as diarrhea, nausea and vomiting.
Also there is the report of the coherent detection research to tranexamic acid in the prior art, such as " in whitening spot-removing class cosmetics
The detection method research of tranexamic acid " in record, it establishes the detection method of tranexamic acid in whitening spot-removing class cosmetics,
Using C18 chromatographic column, it is with 0.23% sodium dodecyl sulfate solution of mass fraction and the eluent of methanol (volume ratio 60/40)
Mobile phase, flow velocity 1.0mL/min, Detection wavelength 220nm carry out high performance liquid chromatography quantitative detection.As a result it detects, ammonia first
Naphthenic acid linear relationship within the scope of 0~1.073mg/mL is good, and detection is limited to 1.45mg/g, average recovery rate 104.36%.
The method is quick and precisely, specificity is strong, high sensitivity, can be used for the detection of tranexamic acid in whitening spot-removing class cosmetics.But the party
Method need to use high performance liquid chromatography, LC-MS instrument and ultra performance liquid chromatography, and there are testing cost height, and operating method is complicated,
The disadvantages of personnel specialty level requirement is higher is unfavorable for the sending of the daily timely testing result of more batches, is unfavorable for enterprise or list
The disadvantages such as position cost control, is unfavorable for promoting on a large scale, and citizen's acceptance is low.
In addition, document: Determination of tranexamic acid content in Waixueting by
Spectrophotometry [J] .J Lanzhou Med Coll (Lanzhou medical college journal), 2000,26 (4): in 20-21, is adopted
Stop main ingredient TA with the external blood of spectrophotometry and carry out assay, as a result, it has been found that having absorption maximum at 466nm, and blank solution
Noiseless, linear relationship is good in 10.0~62.5 μ gmL concentration, and the rate of recovery is up to 100.91%, RSD 1.09%, method
Accurately, quickly, it is easy.But this method poor anti jamming capability has a degree of dependence to environment, and operation fault tolerance rate is low, right
Relatively high in personnel requirement, new hand is not easy to quickly go up hand, and promotion efficiency is poor.
In view of this, the present invention is specifically proposed.
Summary of the invention
The purpose of the present invention is to provide a kind of detection method of tranexamic acid in cosmetics, the detection of the detection method is limited
Degree can achieve very low, as long as the content of tranexamic acid can detecte in 3wt% or more in cosmetics, and it is more existing
The detection means such as the instrument in technology are more quick, low to environmental requirement, and operation accuracy is high, at low cost, are more people institute
Generally receive, it is easy to operate, it is not necessarily to professional's also implementable operation, therefore should be widely promoted and applied.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
The present invention provides a kind of detection methods of tranexamic acid in cosmetics, include the following steps:
(A) reagent A is successively added in sample to be tested, reagent B shakes up, isolated supernatant；
(B) after addition reagent C shakes up layering in supernatant, separation subnatant adds color developing agent, 90 DEG C of heating, and observation is shown
Pornographic condition；
Wherein, the reagent A is at least one of Tween-20, Tween-80, PEG-20000, and the reagent B is
DMSO、DMF、EtOH、CH2Cl2At least one of, the reagent C be water, saturated sodium chloride solution, saturation phosphoric acid hydrogen dimethylamino it is molten
At least one of liquid, saturation phosphoric acid the second light industry bureau potassium solution, saturation disodium phosphate soln, saturation sodium dihydrogen phosphate.
In the prior art, there are many whitening spot-removing class cosmetics on the market in fact directly to add the ingredient containing tranexamic acid
Enter in cosmetics, but due to being the usage except legal indication, not can guarantee drug safety, once cause legal dispute, place
Comprehend very complicated.Tranexamic acid is that clinically common hemostatic, abuse potential cause eye-blurred, headache, dizziness, tired etc.
Central nervous system symptom；Intracranial thrombosis and bleeding excessively can be caused, causes secondary renal plevis inflammation and ureter sludged blood
Obstruction, in addition there are the symptoms such as diarrhea, nausea and vomiting.
Currently, the problem of state food pharmaceuticals administration general bureau does not temporarily use tranexamic acid to whitening spot-removing class cosmetics
It makes concrete regulation, but it is still necessary to reinforce supervising to it, works out corresponding detection method and limit index.
The present invention in order to solve the above technical problems, according in cosmetics now or other raw material add tranexamic acid or
Remain tranexamic acid impurity method, optimize Pretreatment, improve detection limit and detection result, it is established that complete set it is fast
Speed, accurate detection method, to do one's bit in the future to the control of tranexamic acid, and this method itself is easy to operate,
At low cost, operating environment requirement is also relatively low, suitable for more widely being applied.
Preferably as further enforceable scheme, in the step (A), the reagent B is EtOH, CH2Cl2It is mixed
Liquid is closed, by largely practicing discovery when using EtOH, CH2Cl2Mixed liquor when, Monitoring lower-cut can achieve 3%, improve
The sensitivity of detection.
Wherein, in above-mentioned reagent used, it is contemplated that cosmetic material is slightly soluble in ethyl alcohol, not soluble in water and tranexamic acid is easy
It is dissolved in water.If directly being caused certain with extraction with aqueous solution tranexamic acid therein since fat-soluble cosmetics interfered originally
Emulsion is not easy to extract complete.Tranexamic acid in raw material is comprehensively extracted in order to more acurrate, it is necessary to which sample is all or big
It is partly dissolved and is measured.Using the preferable organic reagent of solvability such as dimethylformamide (DMF), dimethyl sulfoxide
(DMSO), methylene chloride, dehydrated alcohol, methylene chloride-dehydrated alcohol (EtOH, CH2Cl2Mixed liquor) tested respectively.
The result shows that single solvent dissolubility is poor, methylene chloride-dehydrated alcohol mixed solution is preferable to its dissolubility.
Preferably, the reagent C is saturated sodium chloride solution.
Since tranexamic acid is soluble easily in water, it is suitble to be extracted with aqueous solution, it is molten that water, saturated sodium-chloride is then respectively adopted
Liquid, potassium dihydrogen phosphate buffer salt solution are tested, the results showed that, it should not be layered with water extraction, and easily cause solution muddy.
Saturated sodium chloride solution and the layering of potassium dihydrogen phosphate buffer salt solution are good, and solution is clarified.In view of sodium chloride is low in cost,
It easily obtains, solution rate, which is compared, is comparatively fast conducive to the production of high-volume preparation of reagents, so final preferred sodium chloride saturated solution.
Preferably as further enforceable scheme, in the step (A), the reagent B of addition and the body of sample to be tested
Product is than being (1-3): 1, more preferably volume ratio is 2:1.
Preferably as further enforceable scheme, in the step (A), the reagent B of addition and the body of sample to be tested
Product is than being (1-2): 1, more preferably volume ratio is 1:1.
Preferably, addition reagent B stands 30s or more after shaking up, more preferably standing 40-50s.
Preferably as further enforceable scheme, in the step (B), the reagent C of addition and the body of sample to be tested
Product is than being (1-2): 1, more preferably volume ratio is 1:1.
Preferably in a certain proportion by the control of added reagent, if dosage will affect greatly very much the layering of solution
And clarity, dosage is too small to be likely to be breached ideal extraction detection effect, it is therefore desirable to control within the scope of suitable dosage.
Preferably, addition reagent C stands 30s or more after shaking up, more preferably standing 40-50s.
Preferably as further enforceable scheme, in the step (B), added color developing agent is that ninhydrin is molten
Liquid.
Preferably as further enforceable scheme, in the step (B), the solvent of the ninhydrin solution is fourth
At least one of alcohol, acetone, methylene chloride, ethyl acetate.
Preferably as further enforceable scheme, in the step (B), the additive amount of the color developing agent is to be measured
The 4-5wt% of sample.
Preferably as further enforceable scheme, in the step (B), add after vibrating 3-5s after color developing agent again
Heating.
Preferably as further enforceable scheme, in the step (B), the temperature of heating is between 92-95 DEG C；
Preferably, the time of heating is 5min or more, is more preferably 6-8min.
Added color developing agent is generally ninhydrin solution, subsequent first to vibrate 3-5s before heating, so that its mixing
More uniformly, the temperature of heating needs to control at 90 DEG C or more, if temperature is too low not to have any colour developing phenomenon, heating when
Between general control observing response phenomenon after 5min or more, heating, show the content of its tranexamic acid lower than one if colourless
Fixed level can be to a certain degree according to the depth of its color if aobvious bluish violet, illustrates that it contains a certain amount of tranexamic acid
Reaction tranexamic acid content height.
Compared with prior art, the invention has the benefit that
(1) due in the prior art without the rapid detection method for tranexamic acid in cosmetics, and tranexamic acid
Limitation do not formulated yet temporarily using standard, the invention belongs to carry out method preparation in advance, belong to and filled up relevant technology
Blank；
(2) key protection point of the invention is to mention the object in target mechanism using addition different kinds of liquid solvents
It takes, from Tween-20 hydrotropy, the anti-interference extraction object of dichloromethane system and sodium chloride saturated solution water washing cleaning is added, often
One step all plays the role of vital, belongs to the key point in method of the invention entire, furthermore involved in method
The operating parameters such as temperature, time it is also most important, be required to control in suitable opereating specification.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Embodiment 1
Detect the tranexamic acid contained by it as follows to liquid make-up:
1. 1ml sample to be tested is taken to be added in centrifuge tube；
2. 1ml reagent A (PEG-20000), 2ml reagent B (CH is added to centrifuge tube2Cl2), it closes the lid, turn upside down 2-
It shakes up for 3 times, is sure not to shake energetically, take out whole supernatant fractions of bottom into another centrifuge tube after standing 30s；
3. 1ml reagent C (water) is added in the centrifuge tube set to above-mentioned clear liquid, close the lid, 2-3 mixing of turning upside down
, it is sure not to shake energetically, stands 30s or so wait there is obvious layering；
4. being placed in new centrifuge tube from subnatant is taken out in above-mentioned centrifuge tube, color developing agent, the addition of color developing agent is added
Amount is that 5wt% is placed in heating water bath 5min in 90 DEG C of water-baths, observing response phenomenon after shaking mixes 3s energetically.
Using above-mentioned detection method, for the liquid make-up standard sample (sampling amount of different tranexamic acid contents
Be 1ml) testing result it is as shown in table 1 below:
1 testing result of table
| Tranexamic acid content | Develop the color phenomenon | Colour developing degree |
| 3% | It is colourless | - |
| 4% | It is colourless | - |
| 5% | It is colourless to become bluish purple | ++ |
| 6% | It is colourless to become bluish violet | +++ |
| 7% | It is colourless to become deep blue purple color | ++++ |
| 8% | It is colourless to become profound bluish violet | +++++ |
Embodiment 2
Carry out detecting the tranexamic acid contained by it as follows to liquid make-up:
1. 1ml sample to be tested is taken to be added in centrifuge tube；
2. 2ml reagent A (Tween-20), 2ml reagent B (EtOH) is added to centrifuge tube, closes the lid, turn upside down 2-3 times
It shakes up, is sure not to shake energetically, take out whole supernatant fractions of bottom into another centrifuge tube after standing 40s；
3. 2ml reagent C (saturation phosphoric acid hydrogen dimethylamino solution) is added in the centrifuge tube set to above-mentioned clear liquid, close the lid,
It turns upside down 2-3 times and mixes, be sure not to shake energetically, stand 40s or so wait there is obvious layering；
4. being placed in new centrifuge tube from subnatant is taken out in above-mentioned centrifuge tube, color developing agent, the addition of color developing agent is added
Amount is that 4wt% is placed in heating water bath 6min in 92 DEG C of water-baths, observing response phenomenon after shaking mixes 5s energetically.
Using above-mentioned detection method, for the liquid make-up standard sample (sampling amount of different tranexamic acid contents
Be 1ml) testing result it is as shown in table 2 below:
2 testing result of table
| Tranexamic acid content | Develop the color phenomenon | Colour developing degree |
| 3% | It is colourless | - |
| 4% | It is colourless | - |
| 5% | It is colourless to become bluish purple | ++ |
| 6% | It is colourless to become bluish violet | +++ |
| 7% | It is colourless to become deep blue purple color | ++++ |
| 8% | It is colourless to become profound bluish violet | +++++ |
Embodiment 3
Carry out detecting the tranexamic acid contained by it as follows to liquid make-up:
1. 1ml sample to be tested is taken to be added in centrifuge tube；
2. 3ml reagent A (Tween-20), 1.5ml reagent B (EtOH+CH is added to centrifuge tube2Cl2), it closes the lid, up and down
It overturns 2-3 times and shakes up, be sure not to shake energetically, take out whole supernatant fractions of bottom into another centrifuge tube after standing 50s；
3. 1.5ml reagent C (saturated sodium chloride solution) is added in the centrifuge tube set to above-mentioned clear liquid, close the lid, on
2-3 mixing is overturned down, is sure not to shake energetically, stands 50s or so wait there is obvious layering；
4. being placed in new centrifuge tube from subnatant is taken out in above-mentioned centrifuge tube, color developing agent, the addition of color developing agent is added
Amount is that 4.5wt% is placed in heating water bath 8min in 95 DEG C of water-baths, observing response phenomenon after shaking mixes 4s energetically.
Solid and semisolid cosmetic standard sample using above-mentioned detection method, for different tranexamic acid contents
The testing result of (sampling amount is 1ml) is as shown in table 3 below:
3 testing result of table
| Tranexamic acid content | Develop the color phenomenon | Colour developing degree |
| 3% | It is colourless to become bluish violet | +++ |
| 4% | It is colourless to become deep blue purple color | ++++ |
| 5% | It is colourless to become deep blue purple color | ++++ |
| 6% | It is colourless to become deep blue purple color | ++++ |
| 7% | It is colourless to become deep blue purple color | ++++ |
| 8% | It is colourless to become profound bluish violet | +++++ |
Embodiment 4
Concrete operation step and embodiment 3 are consistent, and only the dosage of reagent B is 0.5ml.
Solid and semisolid cosmetic standard sample using above-mentioned detection method, for different tranexamic acid contents
The testing result of (sampling amount is 1ml) is as shown in table 4 below:
4 testing result of table
| Tranexamic acid content | Develop the color phenomenon | Colour developing degree |
| 3% | It is colourless to become bluish purple | ++ |
| 4% | It is colourless to become bluish violet | +++ |
| 5% | It is colourless to become deep blue purple color | ++++ |
| 6% | It is colourless to become deep blue purple color | ++++ |
| 7% | It is colourless to become deep blue purple color | ++++ |
| 8% | It is colourless to become profound bluish violet | +++++ |
Embodiment 5
Concrete operation step and embodiment 3 are consistent, and only the dosage of reagent C is 0.4ml.
Solid and semisolid cosmetic standard sample using above-mentioned detection method, for different tranexamic acid contents
The testing result of (sampling amount is 1ml) is as shown in table 5 below:
5 testing result of table
| Tranexamic acid content | Develop the color phenomenon | Colour developing degree |
| 3% | It is colourless to become bluish purple | ++ |
| 4% | It is colourless to become bluish violet | +++ |
| 5% | It is colourless to become bluish violet | +++ |
| 6% | It is colourless to become deep blue purple color | ++++ |
| 7% | It is colourless to become deep blue purple color | ++++ |
| 8% | It is colourless to become profound bluish violet | +++++ |
Comparative example 1
Other operating procedures and embodiment 3 are consistent, and the temperature only heated is 80 DEG C, using above-mentioned detection method, needle
It is as shown in table 6 below to the testing result of the liquid make-up standard sample (sampling amount is 1ml) of different tranexamic acid contents:
6 testing result of table
| Tranexamic acid content | Develop the color phenomenon | Colour developing degree |
| 3% | It is colourless | - |
| 4% | It is colourless | - |
| 5% | It is colourless | - |
| 6% | It is colourless | - |
| 7% | It is colourless | - |
| 8% | It is colourless | - |
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from of the invention
Many other change and modification can be made in the case where spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. the detection method of tranexamic acid in cosmetics, which comprises the steps of:
(A) reagent A is successively added in sample to be tested, reagent B shakes up, isolated supernatant；
(B) after addition reagent C shakes up layering in supernatant, separation subnatant adds color developing agent, and the feelings that develop the color are observed in 90 DEG C of heating
Condition；
Wherein, the reagent A be at least one of Tween-20, Tween-80, PEG-20000, the reagent B be DMSO,
DMF、EtOH、CH2Cl2At least one of, the reagent C is water, saturated sodium chloride solution, saturation phosphoric acid hydrogen dimethylamino solution, satisfies
With at least one of phosphoric acid the second light industry bureau potassium solution, saturation disodium phosphate soln, saturation sodium dihydrogen phosphate.
2. detection method according to claim 1, which is characterized in that in the step (A), the reagent B be EtOH,
CH2Cl2Mixed liquor；
Preferably, the reagent C is saturated sodium chloride solution.
3. detection method according to claim 1, which is characterized in that in the step (A), the reagent A of addition with it is to be measured
The volume ratio of sample is (1-3): 1, more preferably volume ratio is 2:1.
4. detection method according to claim 1, which is characterized in that in the step (A), the reagent B of addition with it is to be measured
The volume ratio of sample is (1-2): 1, more preferably volume ratio is 1:1；
Preferably, addition reagent B stands 30s or more after shaking up, more preferably standing 40-50s.
5. detection method according to claim 1, which is characterized in that in the step (B), the reagent C of addition with it is to be measured
The volume ratio of sample is (1-2): 1, more preferably volume ratio is 1:1；
Preferably, addition reagent C stands 30s or more after shaking up, more preferably standing 40-50s.
6. detection method according to claim 1, which is characterized in that in the step (B), added color developing agent is indenes
Triketone solution.
7. detection method according to claim 6, which is characterized in that in the step (B), the ninhydrin solution it is molten
Agent is at least one of butanol, acetone, methylene chloride, ethyl acetate.
8. detection method according to claim 1, which is characterized in that in the step (B), the additive amount of the color developing agent
For the 4-5wt% of sample to be tested.
9. detection method according to claim 1, which is characterized in that in the step (B), vibrate 3- after adding color developing agent
It is reheated after 5s.
10. detection method according to claim 1, which is characterized in that in the step (B), the temperature of heating is 92-95
Between DEG C；
Preferably, the time of heating is 5min or more, is more preferably 6-8min.
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2018
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| CN101810543A (en) * | 2008-10-08 | 2010-08-25 | 莱雅公司 | Cosmetic composition and a light stabilizing method thereof |
| JP2012008044A (en) * | 2010-06-25 | 2012-01-12 | Yamaguchi Univ | Measurement method of platelet activation capacity and anti-platelet drug |
| CN104569187A (en) * | 2014-12-17 | 2015-04-29 | 深圳出入境检验检疫局食品检验检疫技术中心 | Detection method and device for whitening and freckle-removing components in cosmetics |
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