CN103969393A - Pretreatment method for measurement on contents of glycoside Chinese herbal medicine compounds in cosmetics - Google Patents
Pretreatment method for measurement on contents of glycoside Chinese herbal medicine compounds in cosmetics Download PDFInfo
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- CN103969393A CN103969393A CN201410169330.3A CN201410169330A CN103969393A CN 103969393 A CN103969393 A CN 103969393A CN 201410169330 A CN201410169330 A CN 201410169330A CN 103969393 A CN103969393 A CN 103969393A
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Abstract
The invention relates to a pretreatment method for measurement on contents of glycoside Chinese herbal medicine compounds in cosmetics, and the pretreatment method can be applied to water-borne agents, creams and emulsions. The contents of glycoside Chinese herbal medicine compounds are measured by adopting reversed-phase liquid chromatography. The pretreatment method comprises the following steps: using methanol to disperse a cosmetic sample containing glycoside compounds; adding n-hexane for single-stage extraction; discarding the part containing grease, or/and adding precipitants, namely potassium ferrocyanide aqueous solution, and zinc acetate aqueous solution; centrifuging to discard the part containing macromolecular substances; conducting rotary evaporation; fixing the volume for measuring. The pretreatment method has the advantages as follows: the extraction and separation process is simple and convenient; further concentration is facilitated; the cost is low; the recovery rate is high; the applicability is high; the response value of the target peak is greater than that of a common pretreatment method; the pretreatment method can be used for measurement on the contents of low-content glycoside compounds in cosmetics.
Description
Technical field
The present invention relates to the method for cosmetics Chinese herbal medicine compounds content test pre-treatment.
Background technology
Natural cosmetics is the development trend of following cosmetics, and increasing native compound, medicinal herb components become the focus of declaring of cosmetic product gradually.For the detection of medicinal herb components, need to be by pre-treatment to improve recall rate.
With reference in < < cosmetics health standard > > (2007 editions) (hereinafter to be referred as cosmetics health standard) to the pre-treatment of sex hormone and GB GBT24800.2-2009(hereinafter to be referred as GB) in pre-treating method to 41 kinds of glucocorticoids in cream kind cosmetics, conventional pre-treating method is listed in table 1.
Method in GB and cosmetics health standard is all, after using saturated nacl aqueous solution that cream kind cosmetic sample is disperseed in advance, with an organic solvent object to be extracted.
And at present, there is following subject matter in conventional pre-treating method before detecting cosmetics with reversed phase liquid chromatography:
(1) use mobile phase directly to process cosmetic sample, be equivalent to the direct dilution to sample, the detection limit of sample is higher;
(2) use object in the separated cosmetics of solid phase extraction column, sample preparation cost is high, and solid phase pillar treatment capacity is few, causes the result data of a plurality of sample rooms to have certain systematic error.
(3) use saturated aqueous common salt to carry out breakdown of emulsion processing to oil-in-water type cosmetic, with organic solvent, extracting object, cause sample detection limit high, and the cosmetic sample of processing is not suitable for concentration.
In view of the above problems, the existing pre-treating method detecting for cosmetics is needed optimization badly, to meet cosmetics testing requirement.
Summary of the invention
Technical matters to be solved by this invention is to provide glucosides class Chinese herbal medicine compounds content in a kind of cosmetics to test more common pre-treating method.
The present invention solves the problems of the technologies described above taked technical scheme: a kind of pre-treating method for the test of cosmetics glucosides class Chinese herbal medicine compounds content, described glucosides class Chinese herbal medicine compound adopts liquid chromatography for measuring content, and described pre-treating method comprises the steps:
(1) with methyl alcohol, disperse the cosmetic sample of sugary glycoside Chinese herbal medicine compound, add normal hexane single-stage extraction, discard the part that comprises lubricant component, and/or add precipitation agent ferrocyanide aqueous solutions of potassium and aqueous zinc acetate solution, centrifugal, discard the part that comprises macromolecular substances;
(2) rotary evaporation, constant volume, to be measured.
One of key distinction of the present invention and conventional cosmetics pre-treating method is to adopt methyl alcohol to disperse cream kind cosmetic sample, after removing grease and/or macromolecular substances, then heats up in a steamer except methanol solvate.
Use n-hexane extraction, object is to remove fat-soluble stronger grease, fatty acid compound, by removing upper strata liquid after extraction, can discard the part that comprises lubricant component.Because composition in aqua is more single, FAF compounds, therefore without using n-hexane extraction
Add precipitation agent, object is for next step rotary evaporation, to remove sample to disturb, and by the centrifugal macromolecular substances that makes, precipitates, and can discard the part that comprises macromolecular substances.Because water-soluble substances interference in Water-In-Oil system creme is less, therefore without using precipitation agent.
But do not get rid of the pre-treating method that various formulations all adopt n-hexane extraction to combine with precipitation agent.
Finally, from different as dispersion solvent with saturated sodium-chloride water solution, adopt methyl alcohol as solvent, be convenient to, by rotary evaporation, sample is carried out to concentration, reduced the detection limit of glucoside compound in cosmetics.
Described cosmetics matrix is aqua, creme (Water-In-Oil system) or emulsion (oil-in-water system).
In step (1), for creme or emulsion, first use a small amount of methyl alcohol dispersed sample, then add surplus methyl alcohol, ultrasound wave is assisted breakdown of emulsion; For aqua, first use a small amount of methyl alcohol dispersed sample, after adding precipitation agent, add again surplus methyl alcohol, ultrasound wave is assisted breakdown of emulsion, and ultrasonic time is 5～12min.
Take cosmetic sample quality as 2～5 g, and the addition of methyl alcohol is 5～30 mL, preferably 10～25 mL.
Take cosmetic sample quality as 2～5 g, the addition of ferrocyanide aqueous solutions of potassium and aqueous zinc acetate solution is respectively 10～100 μ L, preferred 20～50 μ L, the concentration of ferrocyanide aqueous solutions of potassium is 5～20 wt%, preferred 8～15 wt%, the concentration of aqueous zinc acetate solution is 10～30 wt%, preferably 15～25 wt%.
Precipitation agent concentration can regulate according to addition.Before adding precipitation agent, also suitable quantity of water can be added so that precipitation agent plays a role.
Take cosmetic sample quality as 2～5 g, and the addition of normal hexane is 10～30 mL, preferably 15～25 mL.
Add after normal hexane ultrasound wave auxiliary extraction 3～8 minutes.
In step (2), rotary evaporation carries out under vacuum and water bath condition, and bath temperature is 55～65 ℃.
Rotary evaporation is by the basic evaporate to dryness of dispersion solvent methyl alcohol, then mobile phase used carries out constant volume while measuring by reverse phase liquid chromatography method.After constant volume, also can be through 0.45 μ m membrane filtration.
Described glucosides class Chinese herbal medicine compound comprises rhodioside, ginsenoside, Paeoniflorin, asiaticosid, madecassoside.
The invention has the beneficial effects as follows:
The pre-treating method of compound in extraction cosmetics provided by the invention, extraction detachment process is easy, be convenient to further concentrated, cost is low, the recovery is high, and applicability is good, compared to common pre-treating method, the response of target peak is larger, can be used for measuring the glucoside compound of low content in cosmetics.
Accompanying drawing explanation
The contrast colors spectrogram of Fig. 1 rhodioside aqua sample, wherein, curve 1 is the chromatogram without the sample of pre-treatment, the chromatogram that curve 2 is the sample that obtains through pre-treating method of the present invention.
Embodiment
Adopt methyl alcohol to dissolve cosmetic sample, ultrasound wave is assisted breakdown of emulsion, lubricant component in normal hexane single-stage extraction cosmetics, coordinate ultrasound wave auxiliary extraction, and/or large molecule precipitation agent precipitation, use the high performance liquid chromatography (HPLC) that is equipped with diode array detector (Photodiode Array Detector, DAD) to carry out content detection to glucoside compound conventional in cosmetics, configured the blank matrix of cosmetics simultaneously.Below in conjunction with specific embodiment, the invention will be further described, but the present invention is not limited to following examples.
By percentage by weight shown in table 2 take each component and routinely technique prepare aqua, emulsion and creme cosmetics, and add glucoside compound in the cosmetics of these formulations, by every g cosmetics (three kinds of formulations), contain 100,200 or 300 μ g glucoside compounds, altogether nine samples are treated the sample of pre-treatment.In table 2, listed each component is all buied from the market.
The pre-treating method that in embodiment 1 cosmetics, Determination of Salidroside is measured
Glucoside compound is rhodioside, and rhodioside standard items are purchased from Shanghai one hundred Nian Shidande detection technique company limited, purity >=98%.Structural formula is:
The pre-treating method that Determination of Salidroside is measured, is divided into following three kinds of concrete processing modes according to three kinds of different State of cosmetics:
1, aqua, adopts following method to carry out pre-treatment:
(1) take 5 g samples in 10 mL color-comparison tubes, add a small amount of pure methyl alcohol, on vortex oscillator, be uniformly dispersed, add 50 μ L 10 wt% ferrocyanide aqueous solutions of potassium and 50 μ L 20 wt% aqueous zinc acetate solution, shake up, use pure methanol constant volume to 10 mL, ultrasonic 10min, in the centrifugal 5min of 12000 r/min again;
(2) collect clear liquid to 25mL round-bottomed flask, use vacuum to revolve steaming instrument and revolve steaming (removal methyl alcohol), 60 ℃ of bath temperatures, with mobile phase, dissolve the remnants in flask, be settled to 5 mL, mix constant volume liquid is ultrasonic, in the centrifugal 10min of 12000 r/min, through 0.45 μ m membrane filtration, filtrate for later use, content to be measured.
2, emulsion, adopts following method to carry out pre-treatment:
(1) take 2 g samples in 50mL color-comparison tube, add a small amount of pure methyl alcohol, after being uniformly dispersed on vortex oscillator, by pure methanol constant volume, to 25mL, ultrasonic 10min, adds 20 mL normal hexanes, mix, ultrasonic 5min, removes upper strata liquid, adds 4 mL water, add again 20 μ L10 wt% ferrocyanide aqueous solutions of potassium and 20 μ L 20 wt% aqueous zinc acetate solution, after shaking up, ultrasonic 5min, the centrifugal 5min of 12000 r/min;
(2) collect clear liquid to 25mL round-bottomed flask, use vacuum to revolve steaming instrument and revolve steaming, 60 ℃ of bath temperatures, dissolve the remnants in flask with mobile phase, be settled to 5mL, mix constant volume liquid is ultrasonic, in the centrifugal 10min of 12000 r/min, through 0.45 μ m membrane filtration, filtrate for later use, content to be measured.
3, creme, adopts following method to carry out pre-treatment:
(1) take 2g sample in 50mL color-comparison tube, add a small amount of pure methyl alcohol, after being uniformly dispersed on vortex oscillator, by pure methanol constant volume, to 25mL, ultrasonic 10min, adds 20mL normal hexane, mixes, and ultrasonic 5min, removes upper strata liquid;
(2) collect clear liquid to 25mL round-bottomed flask, use vacuum to revolve to steam instrument and revolve steaming, 60 ℃ of bath temperatures, dissolve the remnants in flask with mobile phase, are settled to 5mL, mix constant volume liquid is ultrasonic, through 0.45 μ m membrane filtration, filtrate for later use, content to be measured.
Chromatographic condition: with being furnished with DAD detecting device, Agilent 1260 high performance liquid chromatographs of octadecylsilane chemically bonded silica post carry out assay to sample, chromatographic condition is with reference to the chromatographic condition in < < Chinese Pharmacopoeia > > (2010 editions Firsts), mobile phase is V (water): V (methyl alcohol)=85:15,25 ℃ of column temperatures, wavelength 225 nm, flow velocity 1.0 mL/min, sample size 20 μ L.
In rhodioside aqua, emulsion or creme, the recovery of rhodioside is in Table 3.
Table 3 shows, after pre-treatment, in sample, detected Determination of Salidroside and its actual content are more approaching, to glucosides addition, are especially the sample of 200 or 300 μ g/g, and the recovery all approaches 100%.
Separately the cosmetics aqua without pre-treatment is carried out finding after identical assay, as shown in Figure 1, without the target peak response of the sample (curve 1) of pre-treatment, be starkly lower than the sample (curve 2) through pre-treatment.
The pre-treating method of measuring content of ginsenoside in embodiment 2 cosmetics
Glucoside compound is ginsenoside Rb1, Rc, Rb2, Rd, and ginsenoside Rb1, Rc, Rb2, Rd standard items are purchased from Shanghai one hundred Nian Shidande detection technique company limited, purity >=98%.Structural formula is:
The specific operation process of the assay pre-treating method of 4 kinds of ginsenosides that this method provides is identical with embodiment 1.
Chromatographic condition: utilize and be furnished with DAD detecting device, Agilent 1260 high performance liquid chromatographs of octadecylsilane chemically bonded silica post carry out assay to sample, chromatographic condition mobile phase is: V (water): V (acetonitrile)=70:30,25 ℃ of column temperatures, wavelength 203 nm, flow velocity 1.0 mL/min, sample size 20 μ L.
In ginsenoside (Rb1, Rc, Rb2, Rd) aqua, emulsion or creme, the recovery (in total saponin(e) of ginsenoside is in Table 4.
Table 4 shows, after pre-treatment, in sample, detected content of ginsenoside and its actual content are more approaching, and except glucosides addition is the aqua or samples of latex of 100 μ g/g, the recovery all approaches 100%.
The pre-treating method that in embodiment 3 cosmetics, paeoniflorin content is measured
Glucoside compound is Paeoniflorin, and Paeoniflorin standard items are purchased from Shanghai one hundred Nian Shidande detection technique company limited, purity >=98%.Structural formula is:
The specific operation process of the assay pre-treating method of the Paeoniflorin that this method provides is identical with example 1.
Chromatographic condition: utilize and be furnished with DAD detecting device, Agilent 1260 high performance liquid chromatographs of octadecylsilane chemically bonded silica post carry out assay to extract, chromatographic condition mobile phase is: V (0.1% phosphoric acid solution): V (acetonitrile)=86:14,25 ℃ of column temperatures, wavelength 230 nm, flow velocity 1.0 mL/min, sample size 10 μ L.
In the aqua of Paeoniflorin, emulsion or creme, the recovery of Paeoniflorin is in Table 5.
Table 5 shows, after pre-treatment, in sample, detected paeoniflorin content and its actual content are more approaching, and the recovery all approaches 100%.
The pre-treating method of asiaticosid, madecassoside assay in embodiment 4 cosmetics
Glucoside compound is asiaticosid or madecassoside, and asiaticosid or madecassoside standard items are all purchased from Shanghai one hundred Nian Shidande detection technique company limited, purity >=98%.Structural formula is:
The specific operation process of the assay pre-treating method of the asiaticosid that this method provides, madecassoside is identical with example 1.
Chromatographic condition: utilize and be furnished with DAD detecting device, Agilent 1260 high performance liquid chromatographs of octadecylsilane chemically bonded silica post carry out assay to sample, chromatographic condition mobile phase is V (2 mmol/L beta cyclodextrin solution): V (acetonitrile)=76:24,25 ℃ of column temperatures, wavelength 205 nm, flow velocity 1.0 mL/min, sample size 10 μ L.
Asiaticosid in asiaticosid, madecassoside aqua, emulsion and creme, the madecassoside recovery are in Table 6.
Table 6 shows, after pre-treatment, in sample, detected asiaticosid or madecassoside content and its actual content are more approaching, and except asiaticosid addition is the aqua or samples of latex of 100 μ g/g, the recovery all approaches 100%.
Embodiment 5 is containing the pre-treating method of the cosmetics aqua of rhodioside
(1) take 2 g samples in 10 mL color-comparison tubes, add a small amount of pure methyl alcohol, on vortex oscillator, be uniformly dispersed, add 30 μ L 15 wt% ferrocyanide aqueous solutions of potassium and 30 μ L 25 wt% aqueous zinc acetate solution, shake up, use pure methanol constant volume to 10 mL, ultrasonic 10 min, in centrifugal 5 min of 12000 r/min again;
(2) collect clear liquid to 25 mL round-bottomed flask, use vacuum to revolve steaming instrument and revolve steaming (removal methyl alcohol), 60 ℃ of bath temperatures, with mobile phase, dissolve the remnants in flask, be settled to 5 mL, mix constant volume liquid is ultrasonic, in centrifugal 10 min of 12000 r/min, through 0.45 μ m membrane filtration, filtrate for later use, content to be measured.
Embodiment 6 is containing the pre-treating method of the cosmetic emulsion of rhodioside
(1) take 5 g samples in 25 mL color-comparison tubes, add a small amount of pure methyl alcohol, after being uniformly dispersed on vortex oscillator, with pure methanol constant volume to 20 mL, ultrasonic 10 min, add 15 mL normal hexanes, mix, ultrasonic 5 min, remove upper strata liquid, add 2 mL water, add again 60 μ L 7.5 wt% ferrocyanide aqueous solutions of potassium and 60 μ L 10 wt% aqueous zinc acetate solution, after shaking up, ultrasonic 5 min, centrifugal 5 min of 12000 r/min;
(2) collect clear liquid to 25 mL round-bottomed flask, use vacuum to revolve steaming instrument and revolve steaming, 60 ℃ of bath temperatures, dissolve the remnants in flask with mobile phase, be settled to 5 mL, mix constant volume liquid is ultrasonic, in centrifugal 10 min of 12000 r/min, through 0.45 μ m membrane filtration, filtrate for later use, content to be measured.
Embodiment 7 is containing the pre-treating method of the cosmetics creme of Paeoniflorin
(1) take 3 g samples in 25 mL color-comparison tubes, add a small amount of pure methyl alcohol, after being uniformly dispersed, with pure methanol constant volume to 15 mL, ultrasonic 10 min, add 15 mL normal hexanes on vortex oscillator, mix, ultrasonic 5 min, remove upper strata liquid;
(2) collect in clear liquid to 25 mL round-bottomed flask, use vacuum to revolve to steam instrument and revolve steaming, 60 ℃ of bath temperatures, dissolve the remnants in flask with mobile phase, are settled to 5 mL, mix constant volume liquid is ultrasonic, through 0.45 μ m membrane filtration, filtrate for later use, content to be measured.
Embodiment 8 is containing the pre-treating method of the cosmetics creme of asiaticosid
(1) take 2 g samples in 25 mL color-comparison tubes, add a small amount of pure methyl alcohol, after being uniformly dispersed, with pure methanol constant volume to 16 mL, ultrasonic 10 min, add 25 mL normal hexanes on vortex oscillator, mix, ultrasonic 5 min, remove upper strata liquid;
(2) collect in clear liquid to 25 mL round-bottomed flask, use vacuum to revolve to steam instrument and revolve steaming, 60 ℃ of bath temperatures, dissolve the remnants in flask with mobile phase, are settled to 5 mL, mix constant volume liquid is ultrasonic, through 0.45 μ m membrane filtration, filtrate for later use, content to be measured.
Claims (9)
1. for a pre-treating method for cosmetics glucosides class Chinese herbal medicine compounds content test, described glucosides class Chinese herbal medicine compound adopts reverse phase liquid chromatography method to measure content, it is characterized in that: described pre-treating method comprises the steps:
(1) with methyl alcohol, disperse the cosmetic sample containing glucoside compound, add normal hexane single-stage extraction, discard the part that comprises lubricant component, and/or add precipitation agent ferrocyanide aqueous solutions of potassium and aqueous zinc acetate solution, centrifugal, discard the part that comprises macromolecular substances;
(2) rotary evaporation, constant volume, to be measured.
2. pre-treating method according to claim 1, is characterized in that: described cosmetic sample is aqua, creme or emulsion.
3. pre-treating method according to claim 2, is characterized in that: in step (1), for creme or emulsion, first use a small amount of methyl alcohol dispersed sample, then add surplus methyl alcohol, ultrasound wave is assisted breakdown of emulsion; For aqua, first use a small amount of methyl alcohol dispersed sample, after adding precipitation agent, add again surplus methyl alcohol, ultrasound wave is assisted breakdown of emulsion, and ultrasonic time is 5～12min.
4. pre-treating method according to claim 1, is characterized in that: take cosmetic sample quality as 2～5 g, the addition of methyl alcohol is 5～30 mL.
5. according to the pre-treating method described in claim 1 or 3, it is characterized in that: take cosmetic sample quality as 2～5 g, the addition of ferrocyanide aqueous solutions of potassium and aqueous zinc acetate solution is respectively 10～100 μ L, the concentration of ferrocyanide aqueous solutions of potassium is 5～20 wt%, and the concentration of aqueous zinc acetate solution is 10～30 wt%.
6. pre-treating method according to claim 1, is characterized in that: take cosmetic sample quality as 2～5 g, the addition of normal hexane is 10～30 mL.
7. pre-treating method according to claim 6, is characterized in that: add after normal hexane ultrasound wave auxiliary extraction 3～8 minutes.
8. pre-treating method according to claim 1, is characterized in that: in step (2), rotary evaporation carries out under vacuum and water bath condition, and bath temperature is 55～65 ℃.
9. pre-treating method according to claim 1, is characterized in that: described glucosides class Chinese herbal medicine compound comprises rhodioside, ginsenoside, Paeoniflorin, asiaticosid, madecassoside.
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Application publication date: 20140806 |