CN105866292B - A kind of method of the homogeneous liquid-liquid extraction of ionic liquid-high effective liquid chromatography for measuring steroid hormone - Google Patents
A kind of method of the homogeneous liquid-liquid extraction of ionic liquid-high effective liquid chromatography for measuring steroid hormone Download PDFInfo
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- G—PHYSICS
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
The invention discloses a kind of analysis methods for establishing 6 kinds of steroid hormones in the homogeneous liquid-liquid extraction of ionic liquid-high effective liquid chromatography for measuring milk and milk products.It is tested by certain density recovery of standard addition, optimizes the condition of the homogeneous liquid-liquid extraction of ionic liquid and the condition of high performance liquid chromatograph.The factor for influencing extraction and sample preparation condition is optimized, the optimum condition of optimization is: 70 μ L ionic liquid 1- hexyl -3- methyl imidazolium tetrafluoroborate ionic liquids make extractant, 80 μ L ionic liquid 1- hexyl -3- methyl imidazolium tetrafluoroborate ionic liquids make extractant, the NH4PF6 of 0.3593g/mL is ion-pairing agent, extract 10s, it is centrifuged 5min, the pH value for adjusting sample solution is 3.0~5.0, NaCl concentration is that 7% this method is easy, quick, environmentally friendly, is suitble to the measurement of 6 kinds of steroid hormones in milk and milk products.
Description
Technical field
The present invention relates to steroid hormones in the homogeneous liquid-liquid extraction of ionic liquid-high effective liquid chromatography for measuring milk and milk products
Analysis method.
Background technique
Steroid hormone is a kind of lipophilic, low molecular weight, active compound, has been widely used for animal husbandry
To achieve the purpose that promote milk animal growth and improve feed efficiency, lead to hormone residues problem in milk and milk products.Due to
Hormone residues may be detrimental to health, and such as lead to tumour relevant to endocrine, birth and birth defects；Induce system genitale
System canceration or leukaemia；Baby and teen-age growth and development can also be impacted, cause sex premature etc..Therefore, European Union and
China forbids using hormone in animal food production.Milk and milk products are as the important composition portion in public ordinary meal
Point, hormone residues problem becomes hot spot concerned by people.Steroid hormone is conveyed by blood, mammary gland is synthesized, and is eventually introduced
Into milk.It is reported that about 60% to 80% estrogen is from milk and dairy products in the diet in west.Based on these changes
The importance and harmfulness for closing object establish a kind of hormone residues in quick, efficient, sensitive method detection milk and milk products extremely
It closes important.
Purification to complex sample matrix and be to introduce the sample into the extraction and concentration of trace target analytes therein
Important link before instrument analysis.Currently, document mainly has liquid-liquid extraction (Liquid- to the report of the pre-treating method of hormone
Liquid extraction LLE, Solid Phase Extraction (Solid phase Extracrion, SPE), supercritical fluid extraction
(Superitical Fluid Extraetion, SFE), refrigerated centrifuge grease removal, accelerated solvent extraction (Accelerated
Solvent ex tract ion, ASE), matrix solid phase dispersion extract (matrix solid-phase dispersion
MSPD), ultrasonic extraction method (ultrasonic extraction, ULE), microwave auxiliary extraction (Microwave-assisted
Extraction, MAE) [, liquid-phase micro-extraction (Liquid phase microextraction, LPME), solid phase microextraction
(Solid phase microextraction), gel infiltration purification chromatography (Gel Permeation Chromatography,
GPC), immuno absorbence extracts (Immunosorbent extration), Stir Bar Sorptive Extraction (Stir Bar Sorptive
Extraction, SBSE) the methods of [.Refrigerated centrifuge grease removal, liquid-phase extraction, Solid Phase Extraction, ultrasonic extraction, microwave auxiliary extraction
Etc. conventional methods generate time it is more early, Dispersive solid phase extraction, supercritical fluid extraction, accelerated solvent extraction, gel seep
Purification chromatography etc. is also applied thoroughly, but in recent years more popular still micro-extraction technique, such as liquid-phase micro-extraction, solid phase microextraction
Equal micro-extraction techniques, or using these micro-extraction techniques in conjunction with traditional extracting process to improve extraction efficiency.
Ionic liquid as a kind of emerging green solvent, have that steam forces down, thermal stability is good, structure is adjustable and
There are the unique physicochemical properties such as excellent dissolution ability to organic and inorganic matter, traditional organic solvent can be replaced, in chemistry
The field homogeneous liquid-liquid extraction of ionic liquid of being used widely is a kind of new micro-extracting method, and principle is by hydrophilic ionic liquid
Body is added in sample solution, adds ion-pairing agent, hydrophobic ionic liquid, targeted are equably generated in sample solution
Close object simultaneously be extracted, be enriched in this newly-generated hydrophobic ionic liquid phase, thus make object reach extraction, separation,
The effect of enrichment.
Document mainly has colorimetric method, thin-layered chromatography (TLC), immunoassay, life to the report of the measuring method of hormone
Object measuring method, gas chromatography (GC), high liquid chromatography method (HPLC), gas chromatography-mass spectrography (GC-MS), gas-chromatography-
Tandem mass spectrometry (GC-MS/MS), HPLC MS (HPLC-MS), high performance liquid chromatography-tandem mass method
(HPLC-MS/MS) etc..Colorimetric method, thin-layered chromatography remolding sensitivity are lower, it is difficult to meet the requirement of quantitative limit in modern analysis,
It is gradually eliminated；Detection of the immunoassay generally just for a certain hormonal components, there is its limitation；Bioassary method is surveyed
Determine process complexity, is also unfavorable for largely promoting and applying；Gas chromatography method is not directly applicable the measurement of hormone, because swashing
The boiling point of element is higher, needs to perform the derivatization laggard gas chromatography measurement, and generally require and analyzed with mass spectrometry,
Derivatization process increases experimental procedure, operates more complicated；High performance liquid chromatography uv detection method is that laboratory is easier
The method of realization is suitble to analysis boiling point high or the substance of thermal stability difference, thus is also suitble to the separation determination of hormone, but sensitivity
It is more weaker compared with HPLC-MS or HPLC-MS/MS with selectivity, but HPLC-MS or HPLC-MS/MS also have the disadvantage of its own,
Costly such as instrument, maintenance cost is higher and needs the higher operator of technical level, and there is also matrix effects in experiment
It should wait.
Summary of the invention
The technical problem to be solved by the present invention is to overcome the existing defects, provides a kind of homogeneous liquid liquid extraction of ionic liquid
Take-high effective liquid chromatography for measuring milk and milk products in 6 kinds of steroid hormones analysis method.
6 kinds of hormone standard items: Mo Meitasong furoate (CAS:83919-23-7), halcinonide (CAS:3039-35-4),
Progesterone (CAS:57-83-0), testosterone propionate (CAS:57-85-2), testobolin (CAS:7207-92-3), hydroxyprogesterone acetate
(CAS:302-23-8).
In order to solve the above-mentioned technical problems, the present invention provides the following technical solutions:
The preparation of standard reserving solution: suitable hormone standard items are accurately weighed respectively, is dissolved and is fully transferred to acetonitrile
In 50.00mL volumetric flask, with dilution in acetonitrile and it is settled to scale, 1000.00 μ g/mL standard reserving solutions are configured to respectively, in -18
DEG C freezing is kept in dark place.
The preparation of hybrid standard intermediate fluid: each standard reserving solution 1.00mL is pipetted respectively in 50.00mL volumetric flask, uses second
Nitrile dilution and constant volume, are configured to the hybrid standard intermediate fluid that each measured object concentration is 20.00 μ g/mL, are protected from light in -4 DEG C of refrigerations
It saves.
The preparation of hybrid standard working solution: being as needed diluted hybrid standard intermediate fluid when use, dilute with acetonitrile
It is interpreted into the hybrid standard working solution of various concentration, matching while using.
Instrument operating condition
High-efficient liquid phase chromatogram condition: C18 chromatographic column (4.6mm*250mm, 5 μm)；Flow velocity 1.0mL/min；Column temperature: 40 DEG C；
Sampling volume: 20 μ L；Detection wavelength: 240nm；Mobile phase A is methanol, and B is water, gradient condition are as follows: 0-2min, 45%A；2-
7min, 45-55%A；7-10min, 55-65%A；10-13min, 65-90%A；13-18min, 90%A；18-18.1min,
90-45%A；18.1-25min 45%A.
The key of the homogeneous liquid-liquid extraction of ionic liquid is the suitable extractant of selection and ion-pairing agent, it is however generally that, it needs
It wants water-soluble preferable ionic liquid as extractant, ion-pairing agent is added, the ionic liquid formed after exchange reaction occurs
Hydrophobicity will be got well, and want high to the extraction yield of object.Experimental selection [C6MIM] [BF4] be used as extractant, NH4PF6 be used as from
Son is to reagent.When also studied [C4MIM] [BF4], [C8MIM] [BF4], [C6MIM] Br plasma liquid as extractant,
Rate of recovery when NH4PF6 is as ion-pairing agent, not as good as the rate of recovery of [C6MIM] [BF4] as extractant.Therefore, finally
Select [C6MIM] [BF4] and NH4PF6 respectively as extractant and ion-pairing agent.
Fixed ion examines 70 μ L, 80 μ L, 90 μ L, 100 μ are added respectively to when the quality of reagent N H4PF6 is 0.35g
L, when 110 μ L, 120 μ L [C6MIM] [BF4] 6 kinds of steroid hormones the rate of recovery.When [C6MIM] [BF4] additional amount is 70 μ L, 6 kinds
The peak area of steroid hormone is relatively large, and with the increase of ionic liquid, the rate of recovery is gradually reduced.
The molar ratio that [C6MIM] [BF4] and NH4PF6 are examined in experiment is respectively 1:1,1:2,1:3,1:4,1:5,1:
6, influence when 1:7,1:8 to experimental result.The result shows that the molar ratio of [C6MIM] [BF4] and NH4PF6 are 1:1 and 1:2
When, the rate of recovery of 6 kinds of steroid hormones is poor, when the molar ratio of [C6MIM] [BF4] and NH4PF6 are lower than 1:7, with NH4PF6 content
Increase, [C6MIM] [PF6] volume increases；When the molar ratio of [C6MIM] [BF4] and NH4PF6 reach 1:7, [C6MIM]
[PF6] volume reaches maximum；After the molar ratio of [C6MIM] [BF4] and NH4PF6 are greater than 1:7, [C6MIM] [PF6] volume base
This is constant, adds the waste that NH4PF6 causes reagent.So the molar ratio for selecting [C6MIM] [BF4] and NH4PF6 is 1:7,
When [C6MIM] [BF4] of 70 μ L is added, the quality of the NH4PF6 of addition is 0.3593g.
Extraction time is all a very important factor for influencing extraction efficiency either in the extraction of what form.
Extraction time is too short, and distribution of the object in two-phase may not be able to reach balance；Extraction time is too long, but increase the time at
This.But in the homogeneous liquid-liquid extraction of ionic liquid, in the forming process of hydrophobic ionic liquid [C6MIM] [PF6], object
It is come into full contact in aqueous solution with ionic liquid, therefore, even extraction time is very short, and object is also rapidly by water phase point
It is assigned to ionic liquid and mutually reaches balance, to complete extraction process.This be also one of the homogeneous liquid-liquid extraction of ionic liquid especially
Prominent advantage.Experiment exam be vortexed the time that concussion is extracted be respectively 10s, 30s, 1.0min, 2.0min,
The rate of recovery of 6 kinds of steroid hormones when 3.0min, 5.0min, the results showed that, time of 6 kinds of steroid hormones after extraction time is 10s
Yield has tended towards stability, therefore final choice is vortexed concussion extraction time as 10s.
Centrifugation is the process for promoting aqueous phase and ionic liquid phase to be separated.Centrifugation time is too short, point of ionic liquid phase
From may not be sufficient, the rate of recovery of target compound is low；But centrifugation time is too long, also increases the time of experiment.It is general and
Speech, low temperature are also beneficial to reduce the solubility of hydrophobic ionic liquid in water, are conducive to the separation of two-phase.Experiment exam -4
Carried out respectively under the conditions of DEG C with 15 000r/min centrifugation 1.0min, 3.0min, 5.0min, 8.0min, 10.0min, 15.0min,
The rate of recovery of 6 kinds of steroid hormones when 20.0min, the results showed that, target when being centrifuged 1.0min, 3.0min, 5.0min under this condition
The rate of recovery difference of object is little, but is centrifuged 5.0min or more the rate of recovery and decreases instead.Therefore, the centrifugation time of 5.0min is
It is enough.Final choice 5.0min is as optimal centrifugation time.
The pH value of sample solution is extremely important in extraction process, because pH value can influence the class of object in the solution
Type, and it is directly related with the rate of recovery.The pH value for examining sample is respectively 1.0,3.0,5.0,6.0,7.0,8.0,10.0,
12.0,14.0 when the rate of recovery, the results showed that, pH value be 5.0 when, the rate of recovery highest of 6 kinds of steroid hormones.PH value be 1.0,
3.0,6.0 when target compound the rate of recovery it is also higher, that is, the rate of recovery is preferable in acid condition for target compound, but
The rate of recovery is substantially reduced some even without recycling under alkaline condition.It may be since this 6 kinds of steroid hormones are faintly acid
Object is closed, under mild acid conditions, object exists in the form of molecule, is easy to form mixture with ionic liquid.In view of cream and
Dairy products go removing protein effect preferable in acid condition, and therefore, the pH value of final choice sample solution is 3.0~5.0.
Salinity is related to ionic strength in 20.00mL sample, this experiment is adjusted with NaCl is added.In general,
With the increase of NaCl, ionic strength increases, and the solubility of the hydrophobic ionic liquid of object and formation in aqueous solution subtracts
It is small, be conducive to the increase of the rate of recovery, i.e. salting out is obvious.But when NaCl concentration is excessively high, Cl- will lead to [C6MIM] again
The generation of [Cl], so that the formation of hydrophobic ionic liquid [C6MIM] [PF6] is influenced, to influence the rate of recovery.Experiment exam
The recycling of 6 kinds of steroid hormones when NaCl (w/v) is respectively 0.0%, 1.0%, 2.0%, 3.0%, 5.0%, 7.0% and 10.0%
Rate, the results showed that, the rate of recovery difference of 6 kinds of steroid hormones is little when NaCl salinity is 1.0%~7.0%, higher.Because
The addition of NaCl facilitate precipitate milk and milk products in protein, therefore, this experiments experiment final choice NaCl concentration it is dense
Degree is 7.0%, it is contemplated that sample solution concentration is 7.0% after 5.0mL sample is extracted with 15mL NaCl solution, selects preparation
NaCl solution concentration is 100g/L.
Optimum condition is: 70 μ L ionic liquid 1- hexyl -3- methyl imidazolium tetrafluoroborate ionic liquids ([C6MIM]
[BF4]) make extractant, 80 μ L ionic liquid 1- hexyl -3- methyl imidazolium tetrafluoroborate ionic liquids ([C6MIM] [BF4])
Make extractant, the NH4PF6 of 0.3593g/mL is ion-pairing agent, extracts 10s, is centrifuged 5min, the pH value for adjusting sample solution is
3.0~5.0, NaCl concentration 7%.With this condition, the rate of recovery of 6 kinds of steroid hormones is 81~124% in milk and milk products,
Relative standard deviation is 3.2~6.5%, is quantitatively limited to 1.0 μ g/kg.
Beneficial effects of the present invention: this method utilizes the homogeneous liquid-liquid extraction techniques of ionic liquid, establishes milk and milk products
In in 6 steroid hormone measuring method, be measured using liquid chromatograph, it is easy to operate, quickly, and high sensitivity.The party
Method is easy, quick, environmentally friendly, is suitble to the measurement of 6 kinds of steroid hormones in milk and milk products.
Detailed description of the invention
The spectrogram of Fig. 1 standard solution
Specific embodiment
Hereinafter, preferred embodiments of the present invention will be described, it should be understood that preferred embodiment described herein is only used
In the description and interpretation present invention, it is not intended to limit the present invention.
Embodiment 1:
High-efficient liquid phase chromatogram condition: C18 chromatographic column (4.6mm*250mm, 5 μm)；Flow velocity 1.0mL/min；Column temperature: 40 DEG C；
Sampling volume: 20 μ L；Detection wavelength: 240nm；Mobile phase A is methanol, and B is water, gradient condition are as follows: 0-2min, 45%A；2-
7min, 45-55%A；7-10min, 55-65%A；10-13min, 65-90%A；13-18min, 90%A；18-18.1min,
90-45%A；18.1-25min 45%A.
The preparation of sample solution: taking 5.00mL milk sample in the Beckman centrifuge tube of 50mL polytetrafluoroethylene (PTFE), is added
The phosphoric acid solution of the NaCl solution of 10.00mL 100g/L and 150 μ L, ultrasonic extraction 5min.With 15 000r/ under the conditions of -4 DEG C
Min is centrifuged 15min, supernatant is transferred in the Beckman centrifuge tube of another 50mL polytetrafluoroethylene (PTFE).It adds
For the NaCl solution of 5.00mL100g/L in residue, 2min is extracted in ultrasound vibration, is centrifuged under the conditions of -4 DEG C with 15 000r/min
15min.Merge supernatant and crosses 0.22 μm of water phase filter membrane as sample solution.
Equal phase extraction process: 70 μ L ionic liquid [C6MIM] [BF4] Yu Shangshu sample solutions are added with micro syringe
In, rear vortex oscillation about 10s mixes well the two, and the solution of homogeneous phase is formed after dissolving ionic liquid all.Then plus
Enter the NH4PF6 aqueous solution of 1.00mL 0.3593g/mL, vortex oscillation 10s, with 15 under the conditions of -4 DEG C after mixing well
000r/min is centrifuged 5.0min, and new hydrophobic ionic liquid is formed and affixed to centrifugation bottom of the tube.After removing upper layer aqueous solution,
100 μ L acetonitriles are added, mixes, is placed in the interpolation pipe of sample injection bottle, is analyzed by high performance liquid chromatograph.All conditions
Optimization experiment is repeated twice.
Using this method to 6 kinds of steroids in plain chocolate, skim milk, lowfat milk, pasteurization milk and superhigh temperature sterilized milk
Body hormone is determined.The result shows that the rate of recovery range of this method is 65.6~115% in actual sample analysis.,
Precision RSD value range is 2.5~6.3%.Progesterone is wherein detected, content is between 1.2~3.8 μ g/kg.Standard solution
Spectrogram such as Fig. 1.
Working curve, detection limit and quantitative limit
Mark-on is carried out to sample solution made of plain chocolate, a series of different mark-on sample of spiked levels is prepared, uses institute
The method of foundation analyzes these mark-on samples.Using spiked levels (c) and peak area (A) as abscissa and ordinate, draw
Working curve processed calculates the equation of linear regression and linearly dependent coefficient of 6 kinds of steroid hormones using liquid chromatograph software automatically,
It is shown in Table 1.
Quantitative limit (Limits of Quantification, LOQ) is when the signal of determinand is 10 with noise (S/N)
The concentration of corresponding determinand.The quantitative limit of 6 kinds of steroid hormones is shown in Table 1.
Precision
In order to examine or check the precision of method, by carrying out 5.0,10.0 and 50.0 μ g/kg these three concentration to plain chocolate
Recovery testu examines the withinday precision and day to day precision of 6 kinds of steroid hormones.Withinday precision be one day within not
The same time carries out 5 experiments (n=5) to 3 spiked levels of sample respectively, examines or check its rate of recovery and relative standard deviation
(RSD).Day to day precision is continuous 5 days (n=5), carries out the experiment of 3 spiked levels to sample daily, examines or check its rate of recovery
And RSD.The withinday precision RSD value of 6 kinds of steroid hormones is located at 3.0~4.5%.Day to day precision RSD value be located at 3.5~
5.8%.
Finally, it should be noted that the foregoing is only a preferred embodiment of the present invention, it is not intended to restrict the invention,
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still may be used
To modify the technical solutions described in the foregoing embodiments or equivalent replacement of some of the technical features.
All within the spirits and principles of the present invention, any modification, equivalent replacement, improvement and so on should be included in of the invention
Within protection scope.
Claims (1)
1. a kind of method of the homogeneous liquid-liquid extraction of ionic liquid-high effective liquid chromatography for measuring steroid hormone, it is characterised in that: packet
Include following steps:
High-efficient liquid phase chromatogram condition: C18 chromatographic column, 4.6mm*250mm, 5 μm；Flow velocity 1.0mL/min；Column temperature: 40 DEG C；Sample introduction body
Product: 20 μ L；Detection wavelength: 240nm；Mobile phase A is methanol, and B is water, gradient condition are as follows: 0-2min, 45%A；2-7min, 45-
55%A；7-10min, 55-65%A；10-13min, 65-90%A；13-18min, 90%A；18-18.1min 90-45%A；
18.1-25min 45%A；
The preparation of sample solution: taking 5.00mL milk sample in the Beckman centrifuge tube of 50mL polytetrafluoroethylene (PTFE), is added
The phosphoric acid solution of the NaCl solution of 10.00mL 100g/L and 150 μ L, ultrasonic extraction 5min, with 15 000r/ under the conditions of -4 DEG C
Min is centrifuged 15min, supernatant is transferred in the Beckman centrifuge tube of another 50mL polytetrafluoroethylene (PTFE)；Add 5.00mL
For the NaCl solution of 100g/L in residue, ultrasonic shake extracts 2min, is centrifuged 15min under the conditions of -4 DEG C with 15 000r/min；Merge
Supernatant crosses 0.22 μm of water phase filter membrane as sample solution；
Equal phase extraction process: being added in 70 μ L ionic liquid [C6MIM] [BF4] Yu Shangshu sample solutions with micro syringe,
Vortex oscillation about 10s afterwards mixes well the two, and the solution of homogeneous phase is formed after dissolving ionic liquid all；Then it is added
The NH4PF6 aqueous solution of 1.00mL 0.3593g/mL, vortex oscillation 10s, with 15 000r/ under the conditions of -4 DEG C after mixing well
Min is centrifuged 5.0min, and new hydrophobic ionic liquid is formed and affixed to centrifugation bottom of the tube；After removing upper layer aqueous solution, it is added
100 μ L acetonitriles mix, are placed in the interpolation pipe of sample injection bottle, are analyzed by high performance liquid chromatograph, steroid hormone Mo Mei
His loose furoate, halcinonide, progesterone, testosterone propionate, testobolin and hydroxyprogesterone acetate.
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| CN106706526B (en) * | 2016-10-12 | 2019-08-20 | 潘仲巍 | Ion liquid abstraction-ultraviolet-visible spectrophotometry measurement mexiletine hydrochloride method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005103700A1 (en) * | 2004-04-21 | 2005-11-03 | The University Of York | Ionic assay medium |
| CN102768245A (en) * | 2011-05-06 | 2012-11-07 | 吉林师范大学 | Method for determining trace chlorophenol endocrine disruptor in water |
| CN102778494A (en) * | 2012-06-01 | 2012-11-14 | 江南大学 | Method for separating and detecting six cortical hormones in skin-care cosmetic |
| CN103822982A (en) * | 2014-02-25 | 2014-05-28 | 浙江大学宁波理工学院 | Method for enriching and detecting phthalate ester compound in wine sample by ionic liquid-dispersive liquid-liquid microextraction |
-
2016
- 2016-05-31 CN CN201610378386.9A patent/CN105866292B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005103700A1 (en) * | 2004-04-21 | 2005-11-03 | The University Of York | Ionic assay medium |
| CN102768245A (en) * | 2011-05-06 | 2012-11-07 | 吉林师范大学 | Method for determining trace chlorophenol endocrine disruptor in water |
| CN102778494A (en) * | 2012-06-01 | 2012-11-14 | 江南大学 | Method for separating and detecting six cortical hormones in skin-care cosmetic |
| CN103822982A (en) * | 2014-02-25 | 2014-05-28 | 浙江大学宁波理工学院 | Method for enriching and detecting phthalate ester compound in wine sample by ionic liquid-dispersive liquid-liquid microextraction |
Non-Patent Citations (4)
| Title |
|---|
| Extraction and determination of hormones in cosmetics by homogeneous ionic liquid microextraction high-performance liquid chromatography;Mingqin Kang 等;《J. Sep. Sci.》;20120816;第35卷(第16期);第2032-2039页 |
| 环境水样中己烯雌酚与双烯雌酚的离子液体分散液-液微萃取/高效液相色谱法测定;张剑 等;《分析测试学报》;20100630;第29卷(第6期);第629-632页 |
| 离子液体液相微萃取在化妆品分析中的应用;康明芹;《中国博士学位论文全文数据库 工程科技Ⅰ辑》;20140915(第09期);第B014-124页 |
| 超声辅助离子液体分散液相微萃取-高效液相色谱法测定废水中雌激素的研究;刘美华 等;《分析测试技术与仪器》;20090930;第15卷(第3期);第151-157页 |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2776013C1 (en) * | 2021-07-09 | 2022-07-12 | Федеральное государственное бюджетное учреждение науки "Федеральный исследовательский центр питания, биотехнологии и безопасности пищи" | Method for quantifying the residual content of steroid hormones in fish |
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