CN102735768B - Process for jointly detecting estrogens and their associations in livestock and poultry excrements - Google Patents
Process for jointly detecting estrogens and their associations in livestock and poultry excrements Download PDFInfo
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Abstract
The invention relates to a technology for detecting endocrine disrupters in the environment, and especially relates to a technology for quantitatively analyzing estrone (E1), 17beta-estradiol (E2), estriol (E3), 17alpha-ethinyl estradiol (EE2), estrone-3-glucuronide (E1-3G), 17beta-estradiol-3-glucuronide (E2-3G), estrone-3-sulfate (E1-3S) and 17beta-estradiol-3-sulfate (E2-3S) in a complex matrix livestock and poultry excrement sample through adopting a liquid chromatography-tandem mass spectrometry technique. The technology comprises the following steps: extracting estrogens in the collected livestock and poultry excrement sample through adopting a rapid solvent extraction method, pre-purifying the resulting extract through liquid-liquid extraction and alkaline solution extraction, enriching targets with an HLB column, purifying estrogen monomers and estrogen associations through using a Florisil column and an NH2 column respectively, and finally detecting through adopting the liquid chromatography-tandem mass spectrometry technique. The process has the advantages of environmental pollution, easy operation, and high recovery rate, and allows the estrogens and their associations having trace amounts in the complex matrix livestock and poultry excrements to be rapidly analyzed.
Description
Technical field
The present invention relates to liquid chromatography-tandem mass spectrometry coupling detection technique field, particularly relate to extraction, purification and the instrument detecting method of estrogen in the complicated feces of livestock and poultry sample and combination thereof.
Background technology
In recent years, natural and synthetic estrogen oestrone (Estrone, E1), 17 beta estradiols (17 β-Estradiol, E2), estriol (Estriol, E3) and 17 α-ethinyl estradiol (17 α-Ethinyl Estradiol EE2) have caused the extensive concern of academia as incretion interferent in the typical environment.Correlation study studies show that the ng/L concentration level can constitute the risk of feminizing to hydrobionts such as fish in the water environment.And the urine and the ight soil that are discharged by people and mammal are the most basic estrogenic sources in the environment, and its discharging is inevitable.Estrogen is via liver metabolism, mainly discharge with fecaluria with glucuronic acid or sulfate combination form, this is comprising oestrone glucosiduronate combination (Estrone-3-glucuronide, E1-3G), 17 beta estradiol glucosiduronate combination (E2-3G, Estradiol-3-glucuronide), oestrone sulfate combination (Estrone-3-sulfate, E1-3S) and 17 beta estradiol sulfate combinations (Estradiol-3-sulfate, E2-3S) etc.Although these estrogen combination non-activities, major part can be decomposed into the estrogen monomer with estrogen active in sewage treatment plant or natural water.Estrogen glucosiduronate combination is reduced to monomer structure easily in environment, and estrogen sulfate combination is difficult for reduction, and the residual concentration in environment is higher relatively.
Along with human living standard's raising, the scale animal and poultry cultivation scale enlarges gradually, as the also increase year by year of feces of livestock and poultry output in the main source of estrogen.Because using of rainfall runoff and agriculture fertilizer, estrogen becomes to the pollution of environment and can not be ignored in the feces of livestock and poultry.Therefore, the concentration of estrogen and combination thereof is significant for the estimation and the source control of such material total emission volumn in investigation and the research different breeding type feces of livestock and poultry.But the feces of livestock and poultry mesostroma is formed extremely complicated, and the estrogen trace exists, and this has proposed higher requirement to sample pre-treatments.
In recent years, the instrument detecting technology of trace organic substance is greatly improved, as gas chromatography-mass spectrography (GC/MS), gas chromatography-tandem mass spectrum coupling (GC/MS/MS), liquid chromatograph mass spectrography (LC/MS), liquid chromatography-tandem mass spectrometry coupling (LC/MS/MS) etc., have higher sensitivity and degree of accuracy.Yet, for the actual environment sample, because the matrix interference effect reduces the recovery of analytical approach and stability greatly.The estrogen that how trace in the feces of livestock and poultry is existed extracts and purifies, and becomes the bottleneck that estrogen is analyzed in the feces of livestock and poultry.Therefore, it is very necessary setting up the estrogen that is applicable to trace existence in Analysis of Complex the host solid sample, particularly feces of livestock and poultry of recovery height, favorable reproducibility and the pre-treating method of combination thereof.
Summary of the invention
The objective of the invention is at estrogen and combination trace in the complex matrices feces of livestock and poultry sample exist, problem such as matrix interference is big, the pretreatment technology difficulty is big, intend a kind of recovery height of exploitation, highly sensitive, analytical technology accurately and rapidly, realization is to estrogen and combination, for example quantitative measurement of E1, E2, E3, EE2, E1-3S, E2-3S, E1-3G, E2-3G.
Technical scheme of the present invention is as follows:
The co-detection method of estrogen and combination thereof in a kind of feces of livestock and poultry sample is characterized in that this method comprises as the lower part:
(1) solid sample extracts
The feces of livestock and poultry sample of gathering is carried out freeze drying, is extractant with acetone/methanol=1: 1 (v/v), with estrogen and the combination in the quick solvent extraction method extraction sample; Rotary evaporation is concentrated into about 1mL then, dries up with nitrogen, and residue gives over to purification.The temperature that quick solvent extraction method of the present invention is selected is 80 ℃, and pressure is 1500psi, static extraction 2 times, each 8min.
(2) preliminary clearning
A. liquid-liquid extraction purifies: with acetonitrile dissolved residue again, add the normal hexane whirlpool again and mix, grease is removed in liquid-liquid extraction, discards upper strata normal hexane phase, repeats twice, and acetonitrile is dried up with nitrogen.
B. solution purification+HLB column purification: the residue after purifying with the NaOH solution ultrasonic dissolution liquid-liquid extraction of 0.1mol/L, the GF/F glass fiber filter of the Whatman company by 0.7 μ m then, triplicate.Behind filtrate and 100mL ultrapure water, the 6mL methyl alcohol mixing, regulate about pH to 3 with the hydrochloric acid of 4mol/L, utilize the HLB solid-phase extraction column (200mg of solid-phase extraction device activation Waters company, 6mL), the order of activation is: 5mL chromatographically pure ethyl acetate, 5mL chromatographically pure methyl alcohol and 10mL ultrapure water, flow velocity are 1mL/min; Adopt the solid-phase extraction column that has activated that water sample is carried out solid phase extraction concentration.After enrichment is finished, the HLB post is drained, products for further purifies.
(3) sample double purification
The a.NH2 column purification
(500mg 6mL) is connected to dry HLB post below with the solid-phase extraction column adapter to the NH2 post of the Waters company that will activate with methyl alcohol.At first use methanol-eluted fractions HLB post and NH2 post tandem arrangement, be collected in the 10mL glass centrifuge tube, the control flow velocity is 1mL/min, is designated as eluent 1; Use estrogen combination on 2% ammoniacal liquor methanol solution (v/v) the wash-out NH2 post again in 10mL glass centrifuge tube, the control flow velocity is 1mL/min, is designated as eluent 2; Eluent 1 flows down slowly at nitrogen respectively with eluent 2 and dries up.
The b.Florisil column purification
The residue of the eluent 1 after dissolved nitrogen dries up again with normal hexane/methylene chloride=3/1 (v/v) solution, Florisil solid-phase extraction column (500mg by the Waters company that activates with normal hexane in advance then, 6mL), use normal hexane/methylene chloride=3/1 (v/v) drip washing again, use acetone/methylene chloride=2/8 (v/v) to be equipped with for eluant, eluent is eluted in the 10mL glass centrifuge tube of eluent 2 residues at last, then eluent is slowly dried up with nitrogen; The residue methanol constant volume, it is to be measured that whirlpool mixes the nylon filter of crossing 0.22 μ m in the back.
(4) liquid chromatography-tandem mass spectrometry analysis
Liquid chromatography-tandem mass spectrometry coupling instrument is: Agilent 1100 highly effective liquid phase chromatographic systems comprise the quaternary infusion pump, automatic sampler (U.S. Agilent company).3200QTRAP type liquid chromatography/tandem mass spectrometer is furnished with electro-spray ionization source (ESI) and Analyst 1.4.1 data processing software (U.S. Applied Biosystem company).
Chromatographic column: Nova-Pak C18 (3.9mm * 150mm * 4 μ m), Waters company.Ion gun: electro-spray ionization source (ESI), the negative ion mode detects; Ion injection electric :-4500V; Temperature: 450 ℃; Gas 1 (GS1, N in the source
2) pressure: 45psi; Gas 2 (GS2, N in the source
2) pressure: 45psi; Gas curtain gas (N
2) pressure: 20psi; Scan mode is multiple reaction monitoring (MRM); Collision gas (N
2) pressure: Medium.
(5) drafting of typical curve is carried out quantitative measurement with external standard method
The drafting of described external standard method typical curve: utilize the standard substance of accurate weighing estrogen of analytical balance and combination thereof to be dissolved in trifluoroacetic acid aqueous solution, be configured to the standard solution of series concentration, adopt the liquid chromatography-tandem mass spectrometry coupling to analyze, it with concentration respectively horizontal ordinate, peak area is that ordinate returns, obtain typical curve, be used for the amount of working sample analyte.
(6) mensuration of the sample and the recovery
Gather the ight soil of certain livestock and poultry farm, target compound in the 1 pair of feces of livestock and poultry extracts set by step, 2 pairs of extracts carry out preliminary clearning set by step again, 3 pairs of samples carry out double purification set by step then, and then detect with the liquid chromatography-tandem mass spectrometry coupling, and the typical curve that obtains with step 5 relatively, finally obtains the content of estrogen in the feces of livestock and poultry sample to be measured and combination thereof by converting.
Adopt same feces of livestock and poultry sample, the addition of pressing 50ng/g adds standard solution, carries out above-mentioned pre-service and measures estrogen and the content of combination, carries out the recovery according to following formula and calculates:
The R-recovery, %;
C-adds object content in the standard solution feces of livestock and poultry sample, ng/g;
C
0-do not add object content in the standard solution feces of livestock and poultry sample, ng/g.
Wherein: described estrogen and combination thereof are: oestrone (E1), 17 beta estradiols (E2), estriol (E3), 17 α-ethinyl estradiol (EE2), oestrone glucosiduronate combination (E1-3G), 17 beta estradiol glucosiduronate combinations (E2-3G), oestrone sulfate combination (E1-3S) and/or 17 beta estradiol sulfate (E2-3S).
Wherein, the HPLC eluent gradient elution requirement of E1, E2, E3, EE2, E1-3S and E2-3S sees Table 1; The HPLC eluent gradient elution requirement of E1-3G and E2-3G sees Table 2.
Table 1 is measured the eluent gradient condition of E1, E2, E3, EE2, E1-3S and E2-3S
Table 2 is measured E1-3G and E2-3G eluent gradient condition
The invention has the beneficial effects as follows the estrogen that adopts in the quick solvent extraction method extraction feces of livestock and poultry, extract with liquid-liquid extraction and alkali lye again extract is carried out preliminary clearning, use HLB post enrichment object then, with Florisil post and NH2 post estrogen monomer and combination are purified respectively, adopt the liquid chromatography-tandem mass spectrometry coupling to detect at last, detectability is low, has higher sensitivity and degree of accuracy.The invention provides that a kind of detectability is low, favorable reproducibility, highly sensitive, the recovery better, the analytical approach that operation is simple, be applicable to E1, E2, E3, EE2, E1-3S, E2-3S, E1-3G, E2-3G in the Analysis of Complex matrix feces of livestock and poultry sample.The recovery of this method and lowest detectable limit, as shown in table 3:
The recovery of table 3 this method and lowest detectable limit
Description of drawings
Fig. 1 is the chromatogram of 10 μ g/L standard solutions
Chromatogram is followed successively by: Fig. 1 a is that oestrone, Fig. 1 b are that 17 beta estradiols, Fig. 1 c are that estriol, Fig. 1 d are that 17 α-ethinyl estradiol, Fig. 1 e are that oestrone sulfate combination, Fig. 1 f are that 17 beta estradiol sulfate combinations, Fig. 1 g are that oestrone glucosiduronate combination, Fig. 1 h are 17 beta estradiol glucosiduronate combinations;
Fig. 2 is the chromatogram of estrogen and combination in certain livestock and poultry farm chicken manure
Chromatogram is followed successively by: Fig. 2 a is that oestrone, Fig. 2 b are that 17 beta estradiols, Fig. 2 c are that estriol, Fig. 2 d are that 17 α-ethinyl estradiol, Fig. 2 e are that oestrone sulfate combination, Fig. 2 f are that 17 beta estradiol sulfate combinations, Fig. 2 g are that oestrone glucosiduronate combination, Fig. 2 h are 17 beta estradiol glucosiduronate combinations.
Embodiment
Below in conjunction with example and accompanying drawing technical scheme of the present invention is further described
(1) solid sample extracts
The feces of livestock and poultry sample of gathering is carried out freeze drying, accurately take by weighing the 0.5g sample, use 66mL acetone/methanol=1: 1 (v/v), with estrogen and the combination in the quick solvent extraction method extraction sample as extraction solvent; Rotary evaporation is concentrated into about 1mL then, dries up with nitrogen, and residue gives over to purification.The temperature that quick solvent extraction method of the present invention is selected is 80 ℃, and pressure is 1500psi, static extraction 2 times, each 8min.
(2) sample preliminary clearning
A. liquid-liquid extraction purifies: with 5mL acetonitrile dissolved residue again, add 10mL normal hexane whirlpool mixing 5min again, materials such as grease are removed in liquid-liquid extraction, discard upper strata normal hexane phase, repeat twice, and acetonitrile is dried up with nitrogen.
B. solution purification+HLB column purification: with 10mL concentration is residue after the NaOH solution ultrasonic dissolution liquid-liquid extraction of 0.1mol/L purifies, the GF/F glass fiber filter of the Whatman company by 0.7 μ m then, triplicate.Behind filtrate and 100mL ultrapure water, the 6mL methyl alcohol mixing, regulate about pH to 3 with the hydrochloric acid of 4mol/L, utilize the HLB solid-phase extraction column (200mg of solid-phase extraction device activation Waters company, 6mL), the order of activation is: 5mL chromatographically pure ethyl acetate, 5mL chromatographically pure methyl alcohol and 10mL ultrapure water, flow velocity is 1mL/min, adopts the solid-phase extraction column that has activated that water sample is carried out solid phase extraction concentration.After enrichment is finished, the HLB post is drained, products for further purifies.
(3) sample double purification
The a.NH2 column purification
(500mg 6mL) is connected to dry HLB post below with the solid-phase extraction column adapter to the NH2 post of the Waters company that will activate with 5mL methyl alcohol.At first use 6mL methanol-eluted fractions HLB post and NH2 post tandem arrangement, be collected in the 10mL glass centrifuge tube, the control flow velocity is 1mL/min, is designated as eluent 1; Use estrogen combination on the 6mL 2% ammoniacal liquor methanol solution wash-out NH2 post again in 10mL glass centrifuge tube, the control flow velocity is 1mL/min, is designated as eluent 2; Eluent 1 flows down slowly at nitrogen respectively with eluent 2 and dries up.
The b.Florisil column purification
The residue of the eluent 1 after dissolved nitrogen dries up again with 5mL normal hexane/methylene chloride=3/1 (v/v) solution, Florisil solid-phase extraction column (500mg by the Waters company that activates with normal hexane in advance then, 6mL), use 5mL normal hexane/methylene chloride=3/1 (v/v) drip washing again, use 6mL acetone/methylene chloride=2/8 (v/v) to be equipped with for eluant, eluent is eluted in the 10mL glass centrifuge tube of eluent 2 residues at last, then eluent is slowly dried up with nitrogen; Residue 1mL methanol constant volume, it is to be measured that whirlpool mixes the nylon filter of crossing 0.22 μ m in the back.
(4) the liquid chromatography-tandem mass spectrometry coupling is analyzed
The condition that the liquid chromatography-tandem mass spectrometry coupling is selected: Agilent 1100 highly effective liquid phase chromatographic systems comprise the quaternary infusion pump, automatic sampler (U.S. Agilent company); The 3200QTRAP tandem mass spectrometer is furnished with electro-spray ionization source (ESI) and Analyst 1.4.1 data processing software (U.S. Applied Biosystem company).
Chromatographic column: Nova-Pak C18 (3.9mm * 150mm * 4 μ m), Waters company.Ion gun: electro-spray ionization source (ESI), the negative ion mode detects; Ion injection electric :-4500V; Temperature: 450 ℃; Gas 1 (GS1, N in the source
2) pressure: 45psi; Gas 2 (GS2, N
2) pressure: 45psi; Gas curtain gas (N
2) pressure: 20psi; Scan mode is multiple reaction monitoring (MRM); Collision gas (N
2) pressure: Medium.The HPLC eluent gradient elution requirement of E1, E2, E3, EE2, E1-3S and E2-3S sees Table 1; The HPLC eluent gradient elution requirement of E1-3G and E2-3G sees Table 2.
(5) drafting of typical curve is carried out quantitative measurement with external standard method
The drafting of described external standard method typical curve: the standard items that utilize the accurate weighing E1 of analytical balance, E2, E3, EE2, E1-3S, E2-3S, E1-3G, E2-3G, be dissolved in the trifluoroacetic acid aqueous solution, be configured to the standard solution of series concentration, adopt the liquid chromatography-tandem mass spectrometry coupling to analyze, it with concentration respectively horizontal ordinate, peak area is that ordinate returns, and obtains typical curve, is used for the amount of working sample analyte.
(6) mensuration of sample recovery rate
Gather feces of livestock and poultry sample to be measured, 1 pair of sample extracts set by step, 2 carry out preliminary clearning set by step again, 3 carry out double purification set by step, and then detect with the liquid chromatography-tandem mass spectrometry coupling, and with the above-mentioned typical curve that obtains relatively, finally obtain the background value of E1, E2, E3, EE2, E1-3S, E2-3S, E1-3G, E2-3G in the feces of livestock and poultry to be measured by converting.
Adopt same feces of livestock and poultry sample, the addition of pressing 50ng/g adds the mixed standard solution of eight kinds of target compounds, carries out pre-service and measures the content of target compound, carries out the recovery after the background correction value and calculates.
Utilize the present invention to detect the implementation process of estrogen and E1, E2 in certain livestock and poultry farm feces of livestock and poultry sample, E3, EE2, E1-3S, E2-3S, E1-3G, E2-3G:
Gather the fresh feces of livestock and poultry sample of certain livestock and poultry farm, in the brown sampling bottle of the 500mL that sodium azide is housed of packing into clean rapidly and put into the insulation can (＜4 ℃) that has ice cube, take back the laboratory, 1 pair of sample extracts set by step, again set by step 2 and step 3 purify, 4 usefulness liquid chromatography-tandem mass spectrometry couplings detect set by step at last, and with typical curve relatively, finally obtain the content of E1 in the testing sample, E2, E3, EE2, E1-3S, E2-3S, E1-3G, E2-3G by converting.
The content (ng/g) of eight kinds of objects in the actual feces of livestock and poultry of table 3
Annotate: n.d. represents not detect
Claims (1)
1. the co-detection method of estrogen and combination thereof in the feces of livestock and poultry sample is characterized in that this method comprises the steps:
(1) solid sample extracts
The feces of livestock and poultry sample of gathering is carried out freeze drying, accurately take by weighing 0.5 g sample, with 66 mL acetone/methanol=1:1(v/v) as extraction solvent, with estrogen and the combination thereof in the quick solvent extraction method extraction sample; Rotary evaporation is concentrated into about 1 mL then, dries up with nitrogen, and residue gives over to purification; Extraction temperature is 80 ℃, and pressure is 1500 psi, the static extraction 2 times, each 8 min;
(2) extract preliminary clearning
A. liquid-liquid extraction purifies: with 5 mL acetonitriles dissolved residue again, add 10 mL normal hexane whirlpools again and mix 5 min, grease is removed in liquid-liquid extraction, discards upper strata normal hexane phase, repeats twice, and acetonitrile is dried up with nitrogen;
B. solution purification+HLB column purification: with 10 mL concentration is residue after the NaOH solution ultrasonic dissolution liquid-liquid extraction of 0.1 mol/L purifies, the GF/F glass fiber filter of the Whatman company by 0.7 μ m then, triplicate; Behind filtrate and 100 mL ultrapure waters, the 6 mL methyl alcohol mixings, regulate about pH to 3 with the hydrochloric acid of 4 mol/L, utilize the HLB solid-phase extraction column of solid-phase extraction device activation Waters company, the order of activation is: 5 mL chromatographically pure ethyl acetate, 5 mL chromatographically pure methyl alcohol and 10 mL ultrapure waters, flow velocity is 1 mL/min, adopts the solid-phase extraction column that has activated that water sample is carried out solid phase extraction concentration; After enrichment is finished, the HLB post is drained, products for further purifies;
(3) sample double purification
The a.NH2 column purification
To be connected to dry HLB post below with the solid-phase extraction column adapter with the NH2 post that 5 mL methyl alcohol activated; At first use 6 mL methanol-eluted fractions HLB posts and NH2 post tandem arrangement, be collected in the 10 mL glass centrifuge tubes, the control flow velocity is 1 mL/min, is designated as eluent 1; Use estrogen combination on 6 mL, the 2% ammoniacal liquor methanol solution wash-out NH2 post again in 10mL glass centrifuge tube, the control flow velocity is 1 mL/min, is designated as eluent 2; Eluent 1 flows down slowly at nitrogen respectively with eluent 2 and dries up;
The b.Florisil column purification
The residue of the eluent 1 after dissolved nitrogen dries up again with the solution of 5 mL normal hexane/methylene chloride=3/1(v/v), Florisil solid-phase extraction column by the Waters company that activates with normal hexane in advance then, use 5 mL normal hexane/methylene chloride=3/1(v/v) drip washing again, use 6 mL acetone/methylene chloride=, then eluent is slowly dried up with nitrogen at last 2/8(v/v) for eluant, eluent is eluted in the 10 mL glass centrifuge tubes that eluent 2 residues are housed; Afterwards the nylon filter of 0.22 μ m is to be measured excessively with 1 mL methanol constant volume, whirlpool mixing for residue;
(4) the liquid chromatography-tandem mass spectrometry coupling is analyzed
Liquid chromatography-tandem mass spectrometry coupling instrument is: Agilent 1100 highly effective liquid phase chromatographic systems, comprise the quaternary infusion pump, the automatic sampler of U.S. Agilent company, 3200QTRAP type liquid chromatography/tandem mass spectrometer is furnished with electro-spray ionization source (ESI) and Analyst 1.4.1 data processing software; Chromatographic column: Nova-Pak C18,3.9mm * 150mm * 4 μ m, Waters company; Ion gun: electro-spray ionization source (ESI), the negative ion mode detects; Ion injection electric :-4500V; Temperature: 450 ℃; Gas GS1 is N in the source
2, pressure: 45psi; Gas GS2 is N in the source
2, pressure: 45psi; Gas curtain gas is N
2, pressure: 20psi; Scan mode is multiple reaction monitoring (MRM); Collision gas is N
2, pressure: Medium;
(5) drafting of typical curve is carried out quantitative measurement with external standard method
The drafting of described external standard method typical curve: the standard items that utilize accurate weighing estrogen of analytical balance and combination thereof, be dissolved in the trifluoroacetic acid aqueous solution, be configured to the standard solution of series concentration, adopt the liquid chromatography-tandem mass spectrometry coupling to analyze, it with concentration respectively horizontal ordinate, peak area is that ordinate returns, and obtains typical curve, is used for the amount of working sample analyte;
(6) mensuration of sample recovery rate
Gather feces of livestock and poultry sample to be measured, 1 pair of sample extracts set by step, 2 carry out preliminary clearning set by step again, 3 carry out double purification set by step, and then detect with the liquid chromatography-tandem mass spectrometry coupling, and with the above-mentioned typical curve that obtains relatively, finally obtain the value of estrogen in the feces of livestock and poultry to be measured and combination thereof by converting;
Described estrogen is oestrone, 17 beta estradiols, estriol, 17 α-ethinyl estradiol, and described combination is oestrone sulfate combination, 17 beta estradiol sulfate combinations, oestrone glucosiduronate combination, 17 beta estradiol glucosiduronate combinations.
2.The co-detection method of estrogen and combination thereof in the feces of livestock and poultry sample as claimed in claim 1, wherein: the HPLC eluent gradient elution requirement of oestrone, 17 beta estradiols, estriol, 17 α-ethinyl estradiol, oestrone sulfate combination, 17 beta estradiol sulfate combinations is as shown in table 1; The HPLC eluent gradient elution requirement of oestrone glucosiduronate combination, 17 beta estradiol glucosiduronate combinations is as shown in table 2:
Table 1
Table 2
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| CN105572238B (en) * | 2014-10-16 | 2017-11-10 | 无锡市寰创环境科技发展有限公司 | A kind of method of estrogen in detection livestock waste |
| CN105547774B (en) * | 2015-12-08 | 2018-11-06 | 北京市农林科学院 | A method of roxarsone and its catabolite in separation broiler chicken excrement |
| CN106589004A (en) * | 2016-10-12 | 2017-04-26 | 环境保护部华南环境科学研究所 | Treating method for pig manure |
| CN108037226A (en) * | 2017-11-22 | 2018-05-15 | 上海市环境科学研究院 | The method that microwave abstracting-Solid Phase Extraction pre-treatment combination LC-MS technology detects 6 kinds of estrogen of three classes in feces of livestock and poultry at the same time |
| CN109470791A (en) * | 2018-11-29 | 2019-03-15 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | A kind of method and kit of high performance liquid chromatography-tandem mass detection serum estradiol |
| CN109900843B (en) * | 2019-03-26 | 2022-09-27 | 上海市环境科学研究院 | Method for simultaneously detecting 7 conjugated estrogens in livestock and poultry manure by combining microwave extraction-solid phase extraction pretreatment with liquid chromatography-mass spectrometry technology |
| CN109959743A (en) * | 2019-03-27 | 2019-07-02 | 上海市环境科学研究院 | The method that Solid Phase Extraction pre-treatment combination LC-MS technology detects 7 kinds of Conjugated Estrogens in rural sewage simultaneously |
| CN110132707B (en) * | 2019-05-05 | 2020-06-09 | 江南大学 | Pretreatment method for ultrasonic-assisted extraction of multiple steroid hormones in sediment |
| CN110794072A (en) * | 2019-10-14 | 2020-02-14 | 上海药明奥测医疗科技有限公司 | Liquid chromatography tandem mass spectrometry detection method for estrone and estradiol in human serum |
| CN112198265B (en) * | 2020-03-16 | 2023-07-14 | 广州金域医学检验中心有限公司 | Pretreatment method, detection method and kit for simultaneously detecting multiple steroid hormones in blood sample |
| CN112557568B (en) * | 2020-12-21 | 2022-07-22 | 广州金域医学检验集团股份有限公司 | Method for detecting estradiol and estrone |
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| CN102331468A (en) * | 2011-08-03 | 2012-01-25 | 北京师范大学 | Method for testing water deposits or estrogen coalitions in soil |
| CN102435681A (en) * | 2011-09-15 | 2012-05-02 | 北京师范大学 | Preprocessing method suitable for analyzing estrogen and bisphenol A in complex matrix solid sample |
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| CN102331468A (en) * | 2011-08-03 | 2012-01-25 | 北京师范大学 | Method for testing water deposits or estrogen coalitions in soil |
| CN102435681A (en) * | 2011-09-15 | 2012-05-02 | 北京师范大学 | Preprocessing method suitable for analyzing estrogen and bisphenol A in complex matrix solid sample |
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