CN106443019A - Method for rapidly detecting multiple glucocorticoids in cosmetics - Google Patents
Method for rapidly detecting multiple glucocorticoids in cosmetics Download PDFInfo
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- CN106443019A CN106443019A CN201610797768.5A CN201610797768A CN106443019A CN 106443019 A CN106443019 A CN 106443019A CN 201610797768 A CN201610797768 A CN 201610797768A CN 106443019 A CN106443019 A CN 106443019A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
- G01N33/559—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody through a gel, e.g. Ouchterlony technique
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Abstract
The invention discloses a method for rapidly detecting multiple glucocorticoids in cosmetics. Phosphate buffering liquid is used as sample diluent and used for rapidly detecting the multiple glucocorticoids in cosmetics. Every 500 ml of sample diluent is prepared from 2.9 g of Na2HPO4'12H2O, 0.3 g of NaH2PO4'2H2O, 2.4 g of NaCl and the balance water. The sample diluent is used, all kinds of glucocorticoids and other ingredients in cosmetics (water aqua, emulsion and ointment frost agent cosmetics) can be well separated, and a to-be-detected sample solution after dilution can be detected through a common dexamethasone detection card in the market. By means of the sample diluent and the method, the at least 11 kinds of glucocorticoid compounds can be well detected, the method detection limit of the water aqua cosmetics is 1 microgram/g, and the method detection limit of the emulsion and ointment frost agent cosmetics is 2 microgram/g.
Description
Technical field
The invention belongs to analysis of cosmetics field, be specifically related to a kind of for multiple sugar cortical hormones in quick detection cosmetics
The method of element.
Background technology
Glucocorticoid is by class steroid hormone, predominantly a cortisol of zona fasciculata secretion in adrenal cortex
(cortisol), there is the effect of regulation sugar, fat and the biosynthesis of protein and metabolism, also have suppression immune response,
Anti-inflammatory, antitoxin, Antishock function.
Short time uses the cosmetics containing glucocorticoid that skin smooth can be made fine and smooth, ruddy delicate, has preferably
Cosmetic result.Therefore often illegally added to by some bad Cosmetic Manufacture business in all kinds of cosmetics of a multitude of names, give consumption
Person brings serious Health cost.If Long-Time Service contains the cosmetics of this kind of material, hormone will be produced and rely on by skin,
And be difficult to break away from.Thinning of skin, telangiectasis, hair follicle atrophy then can occur after Long-Time Service, once disable, skin is just
Can rubescent, itch, erythema, papule, furfur etc. occur, also can cause osteoporosis, muscular atrophy, growth retardation, induction
Or increase the weight of the various bad reactions such as infection and peptic ulcer, abnormal feeling, metabolic disorder.Therefore, China《Cosmetics safety skill
Art specification》(2015 editions) specify this type of material for disabling composition.
At present, in cosmetics, glucocorticoid detection generally uses liquid chromatography mass combination to be analyzed, complex operation,
Check-Out Time is long, is not suitable for superintendent office, testing agency, the requirement of all kinds of mechanisms field quick detection.
Content of the invention
It is an object of the invention to provide a kind of method for multiple glucocorticoids in quick detection cosmetics.
The technical solution used in the present invention is：
Phosphate buffer in the glucocorticoid of quick detection cosmetics as the application of sample diluting liquid.
Preferably, phosphate buffer is the buffer solution of disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride, and its concentration is respectively
For 0.01-0.03M, 0.003-0.005M and 0.06-0.1M, pH is 7-9.
Preferably, phosphate buffer is the buffer solution of disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride, and its concentration is respectively
For 0.01-0.02M, 0.003-0.0045M and 0.06-0.096M, pH is 7-8.
Preferably, phosphate buffer is the buffer solution of disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride, and its concentration is respectively
For 0.016M, 0.0038M and 0.082M, pH is 7.4.
Preferably, the addition of phosphate buffer is every 0.5-1g cosmetic sample to be measured, adds phosphate buffer
To 10mL.
A kind of sample diluting liquid for glucocorticoid in quick detection cosmetics, contains in every 500ml sample diluting liquid
There is following component：2.9gNa2HPO4·12H2O、0.3gNaH2PO4·2H2O, 2.4gNaCl, excess water.
Preferably, the pH value of this sample diluting liquid is 7.4.
Owing to this sample diluting liquid is cushioning liquid, still can reach 7.4 close to its pH value of solution of concentration or similar formulation,
These are all contained in the application claim limited range close to the solution of concentration or similar formulation.
A kind of kit for glucocorticoid in quick detection cosmetics, comprises dexamethasone detection and blocks and above-mentioned
Sample diluting liquid described in one.
A kind of method for glucocorticoid in quick detection cosmetics, comprises the following steps：
1) sample pretreatment：Take 0.5-1g sample, add sample diluting liquid to 10mL, vibration mixing, obtain detected sample molten
Liquid；Wherein sample diluting liquid is as described in above-mentioned any one.
2) drawing measuring samples solution, dropping 3-5 drips in the well of dexamethasone detection card, reads after 5-10 minute
Result.
Preferably, if detection card T line does not develops the color, judge sample contains glucocorticoid, if detection card T line shows
Look then judges not contain in sample glucocorticoid.
Preferably, glucocorticoid includes hydrocortisone, betamethasone, dexamethasone, dexamethasone acetate, cellulose acetate hydrogen
Change cortisone, cortisone acetate, prednisolone, Econopred, fludrocortisone acetate, prednisone acetate and metacortandracin.
The principle of the colloidal gold strip application Competitive assays immunochromatography of the present invention, the determinand in sample is in flowing
During be combined with the monoclonal antibody specific of colloid gold label, it is suppressed that antibody and the upper determinand of NC film detection line (T)-
The combination of protein conjugate.Dissolving when sample object thing extracts dilution through sample buffer, after solution instills sample well, sample is molten
Glucocorticoid in liquid combines with gold labeling antibody, and then closes the antigen-combining site of glucocorticoid on gold labeling antibody, resistance
Only gold labeling antibody glucocorticoid conjugate on cellulose membrane is combined, and is allowed to develop the color more weak, does not even develop the color；Otherwise, such as sample
Product solution does not has glucocorticoid, then gold glucocorticoid conjugate on cellulose membrane for the labeling antibody can not be stoped to be combined,
Thus colour developing is stronger.
Because determinand is cosmetics, it have aqua, emulsion, cream agent point, its matrix is different, so illegally adding
The glucocorticoid adding is present in cosmetics complex component.Presently commercially available various glucocorticoid detection card and kit thereof
Mostly not having supporting dilution, minority has supporting dilution, but the amount of dilution is less, as 40 test paper are equipped with 50mL dilution
Liquid；Often detect just for one of which glucocorticoid (such as dexamethasone) plus commercially available various detection cards,
So the detection of the illegal various glucocorticoids adding in cosmetics on market cannot be met.
The invention has the beneficial effects as follows：
The sample diluting liquid utilizing the present invention can well be each in cosmetics (including aqua, emulsion, cream agent cosmetics)
Planting glucocorticoid to extract from cosmetics, the detected sample solution obtaining after dilution can be with the ground commonly used on the market
Sai meter Song detection card detects.
Inventive samples dilution and extracting method can well detect following glucocorticoid compound species and include：
Hydrocortisone, betamethasone, dexamethasone, dexamethasone acetate, hydrocortisone acetate, cortisone acetate, prednisolone,
Econopred, fludrocortisone acetate, prednisone acetate, metacortandracin.Wherein water system cosmetics method detection is limited to 1 μ g/
G, breast, the cosmetics method detection of cream system are limited to 2 μ g/g.
By the process of the inventive method, one glucocorticoid detection card is detected simultaneously multiple sugar cortex in cosmetics
Hormone, covers species many, highly sensitive, and the degree of accuracy is high, simple to operate, the detection time is short, it is not necessary to Special Equipment, is suitable for scene
Operation, it is adaptable to superintendent office, testing agency, the requirement of all kinds of mechanisms field quick detection.
Brief description
Fig. 1 is the detection card operation chart for glucocorticoid detection；
Fig. 2 is the detection card result judgement figure for glucocorticoid detection；
Fig. 3 is that this 11 kinds of compounds and positive thereof that quickly method of multiple glucocorticoids is covered in detection cosmetics is shown
Figure, in figure, concentration is the sample liquid concentration being added on detection card；
Fig. 4 is testing result；
Fig. 5 is testing result.
Detailed description of the invention
The principle of the colloidal gold strip application Competitive assays immunochromatography of the present invention, the determinand in sample is in flowing
During be combined with the monoclonal antibody specific of colloid gold label, it is suppressed that antibody and the upper determinand of NC film detection line (T)-
The combination of protein conjugate.Dissolving when sample object thing extracts dilution through sample buffer, after solution instills sample well, sample is molten
Glucocorticoid in liquid combines with gold labeling antibody, and then closes the antigen-combining site of glucocorticoid on gold labeling antibody, resistance
Only gold labeling antibody glucocorticoid conjugate on cellulose membrane is combined, and is allowed to develop the color more weak, does not even develop the color；Otherwise, such as sample
Product solution does not has glucocorticoid, then gold glucocorticoid conjugate on cellulose membrane for the labeling antibody can not be stoped to be combined,
Thus colour developing is relatively strong (see Fig. 1).
Colloidal gold strip used in the present invention is purchased from consumables associated therewith supply company, typically without supporting dilution or join
A small amount of dilution.
Below in conjunction with specific embodiment, the present invention is further illustrated, but is not limited thereto.
Embodiment 1
For the method for glucocorticoid in quick detection cosmetics, comprise the steps：
1. prepare sample diluting liquid
Weigh 2.9g Na2HPO4·12H2O、0.3g NaH2PO4·2H2O, 2.4g NaCl is in 500mL beaker, and add water constant volume
To 500mL scale, fully dissolve and shake up, obtain the sample diluting liquid that pH value is 7.4.
Owing to this sample diluting liquid is cushioning liquid, those of ordinary skill in the art are not before paying creative work
Putting the solution making close to concentration or similar formulation, these are all contained in the application close to the solution of concentration or similar formulation
In claim limited range.
2. sample pretreatment：
Aqua cosmetics：Take 1g sample to add in 10mL sample extraction pipe, be settled to scale with Sample Buffer dilution, fully shake
Be shaken to mixed even after as measuring samples solution.
Breast, cream agent shape product：Take 0.5g sample to add in 10mL sample extraction pipe, be settled to Sample Buffer dilution
Scale, as measuring samples solution after shake well mixing.
3. detection method：
Take packaging bag apart, take out dexamethasone detection card；Drawing measuring samples solution with dropper, vertical dropping 3-5 drips in sample-adding
Kong Zhong.Start timing after sample-adding, after 5 minutes, i.e. can read result；If detection line is inconspicuous with control line colour developing depth contrast, can
After 10 minutes, then carry out result interpretation.
4. result judges：
Negative (-)：T line has colour developing, represents that in sample, glucocorticoid concentration is less than detection limit or does not contains glucocorticoid.
Positive (+)：T line is without colour developing, then it represents that in sample, glucocorticoid concentration is higher than detection limit.
Invalid：Quality Control C line does not occurs, shows that operating process is incorrect or detection card lost efficacy.Can refer to Fig. 2 and carry out result
Judge.
If the result display positive, need to use in laboratory the related further experiment Analysis of official method of country with
Confirmation.
Embodiment 2
The following 11 kinds of classes of compounds of method detection utilizing embodiment 1 include：Hydrocortisone, betamethasone, dexamethasone,
Dexamethasone acetate, hydrocortisone acetate, cortisone acetate, prednisolone, Econopred, fludrocortisone acetate,
Prednisone acetate, metacortandracin.
Accurately weigh each 5.0mg of each standard substance, dissolve with ethanol and be settled to 5mL, obtain 1mg/mL mono-mark storing solution, then
With ethanol stepwise dilution to 10 μ g/mL, obtain standard working solution.
The preparation of glucocorticoid standard liquid：By standard working solution Sample Buffer dilution stepwise dilution to 0.1 μ
G/mL, to be measured.
Detecting step is with embodiment 1.Its result shows, this 11 kinds of compounds all can detect, and its positive map is shown in Fig. 3.
Embodiment 3
The method utilizing embodiment 1 carries out blank sample mark-on test.Weigh breast, cream blank sample 0.5g, water intaking system respectively
Blank sample 1mL, in sample extraction pipe, each parallel takes 11 parts, is separately added into 11 kinds of single mark solution of above-described embodiment 2, obtains
Comprise breast, the cream sample of above-mentioned 11 kinds of glucocorticoids.
It is settled to 10mL by Sample Buffer diluted so that it is concentration is that 0.1 μ g/mL is to be measured.Detecting step is with enforcement
Example 1.Its result shows, this 11 kinds of compound mark-ons, in blank sample, and all can detect the positive, and water system cosmetics method detects
Being limited to 1 μ g/g, breast, the cosmetics method detection of cream system are limited to 2 μ g/g.
Embodiment 4
The method of embodiment 1 sample (being shown in Table 1) to the known positive is utilized to test.Result Fig. 4.
Table 1 positive result of the test
Fig. 4 result shows all can detect the positive, consistent with the result of table 1.
Embodiment 5
Collect 50 batch samples (13 batches of water system cosmetics, 37 batches of breasts, cream agent cosmetics) altogether to verify the present invention, simultaneously with
The result of confirmation method compares, and confirmation uses national standard《41 kinds of sugar skins in GB/T 24800.2-2009 cosmetics
The mensuration liquid chromatography tandem mass spectrometry of matter hormone and TLC》In Liquid Chromatography/Mass Spectrometry, the results are shown in Table 2.
Table 2 screening results and final confirmed results
Table 2 result shows：In addition to 1 batch sample detects false positive, remaining is all consistent with confirmed results, and the accuracy rate of method is
98%, sensitivity is 100%, and specificity is 97.7%, and missing inspection is rate 0.
Comparative example 1
Utilizing the method detection estrogen of embodiment 1, the result of the test of the estrogen standard liquid of 2 μ g/mL is shown in Fig. 5.Can by Fig. 5
Knowing, T line has colour developing, is negative.The visible compound such as estrogen with similar steroid structure, will not cause dry to this method
Disturb.This method is good to glucocorticoids compound specificity.
Comparative example 2
Utilize the buffer solution (buffer solutions of 40 detection one bottle of 50mL of snap-fit) that the detection of commercially available dexamethasone blocks and product is supporting
Carry out contrast experiment.
Being diluted according to the guidance of product description and detecting, result can detect the one of 11 kinds of glucocorticoids or several
Kind.
Proceed by test according to the sample pretreatment step of the embodiment of the present invention 1,11 kinds of glucocorticoids can be detected
One or more, but whole 11 kinds of compounds of the present invention cannot be detected.
Owing to the supporting cushioning liquid of this product is the buffer solutions of every 40 detection snap-fit one bottle of 50mL, therefore buffer solution
Amount is far from the demand of satisfied detection, and the dilution in this method once can prepare 500mL, and preparation method is convenient quickly,
Meet the requirement of all kinds of mechanisms field quick detection.
If additionally, according to the ratio of every 40 detection one bottle of 50mL buffer solutions of snap-fit, i.e. processing 1 part of sample only with about
1.2mL buffer solution, if according to the method for this method, sampling amount being reduced (water system 0.1g, breast system 0.05g), then adds 1mL buffering
Liquid dilutes.Owing to glucocorticoid belongs to illegal adding ingredient in cosmetics, it is considered to the uniformity of sample, sampling amount is very few can
Positive leakage sieve can be caused.And the sampling amount of the inventive method can ensure the illegal missing inspection adding glucocorticoid in cosmetics
Rate is good.
Comparative example 3
The sample diluting liquid of the profit water consumption substitution present invention is tested, and other steps are with embodiment 1.Result can detect 11 kinds of sugar skins
One or more of matter hormone, but whole 11 kinds of compounds of the present invention cannot be detected.
Claims (10)
1. phosphate buffer in the glucocorticoid of quick detection cosmetics as the application of sample diluting liquid.
2. application according to claim 1, it is characterised in that：Phosphate buffer is disodium hydrogen phosphate, sodium dihydrogen phosphate
With the buffer solution of sodium chloride, its concentration is respectively 0.01-0.03M, 0.003-0.005M and 0.06-0.1M, and pH is 7-9.
3. application according to claim 1, it is characterised in that：Phosphate buffer is disodium hydrogen phosphate, sodium dihydrogen phosphate
With the buffer solution of sodium chloride, its concentration is respectively 0.01-0.02M, 0.003-0.0045M and 0.06-0.096M, and pH is 7-8.
4. application according to claim 1, it is characterised in that：The addition of phosphate buffer is every 0.5-1g to be measuredization
Cosmetic sample, addition phosphate buffer to 10mL.
5. the sample diluting liquid for glucocorticoid in quick detection cosmetics, it is characterised in that：Every 500ml sample is dilute
Release and liquid contains following component：2.9g Na2HPO4·12H2O、0.3g NaH2PO4·2H2O, 2.4gNaCl, excess water.
6. sample diluting liquid according to claim 5, it is characterised in that：The pH value of this sample diluting liquid is 7.4.
7. the kit for glucocorticoid in quick detection cosmetics, it is characterised in that：Comprise dexamethasone detection
Card and the sample diluting liquid described in any one of claim 5-6.
8. the method for glucocorticoid in quick detection cosmetics, it is characterised in that comprise the following steps：
1）Sample pretreatment：Take 0.5-1g sample, add sample diluting liquid to 10mL, vibration mixing, obtain treating 2）2）Detection sample
Product solution；Wherein sample diluting liquid is as described in any one of claim 5-6；
2）Drawing measuring samples solution, dropping 3-5 drips in the well of dexamethasone detection card, reads knot after 5-10 minute
Really.
9. method according to claim 8, it is characterised in that：If detection card T line does not develops the color, judge sample contains
Glucocorticoid, if detection card T line colour developing, judges not contain glucocorticoid in sample.
10. method according to claim 8, it is characterised in that：Glucocorticoid include hydrocortisone, betamethasone,
Dexamethasone, dexamethasone acetate, hydrocortisone acetate, cortisone acetate, prednisolone, Econopred, acetic acid fluorine
Tixocortol, prednisone acetate and/or metacortandracin.
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Cited By (4)
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| CN109030838A (en) * | 2018-10-11 | 2018-12-18 | 北京工商大学 | It is a kind of for detecting the colloid gold test paper of dexamethasone |
| CN109211888A (en) * | 2018-09-20 | 2019-01-15 | 北京工商大学 | The rapid detection method of glucocorticoid in a kind of cosmetics |
| CN109293771A (en) * | 2018-10-11 | 2019-02-01 | 北京工商大学 | A kind of method and its special monoclonal antibody detecting dexamethasone |
| CN112213501A (en) * | 2020-09-30 | 2021-01-12 | 中山火炬职业技术学院 | Fluorescent microsphere immunochromatographic test paper for quantitatively detecting dexamethasone and preparation method and detection method thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109211888A (en) * | 2018-09-20 | 2019-01-15 | 北京工商大学 | The rapid detection method of glucocorticoid in a kind of cosmetics |
| CN109030838A (en) * | 2018-10-11 | 2018-12-18 | 北京工商大学 | It is a kind of for detecting the colloid gold test paper of dexamethasone |
| CN109293771A (en) * | 2018-10-11 | 2019-02-01 | 北京工商大学 | A kind of method and its special monoclonal antibody detecting dexamethasone |
| CN112213501A (en) * | 2020-09-30 | 2021-01-12 | 中山火炬职业技术学院 | Fluorescent microsphere immunochromatographic test paper for quantitatively detecting dexamethasone and preparation method and detection method thereof |
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