CN107884490A - Method that is a kind of while detecting a variety of progestational hormone in environment water - Google Patents
Method that is a kind of while detecting a variety of progestational hormone in environment water Download PDFInfo
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- CN107884490A CN107884490A CN201711083232.8A CN201711083232A CN107884490A CN 107884490 A CN107884490 A CN 107884490A CN 201711083232 A CN201711083232 A CN 201711083232A CN 107884490 A CN107884490 A CN 107884490A
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- progestational hormone
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 94
- 229940088597 hormone Drugs 0.000 title claims abstract description 62
- 239000005556 hormone Substances 0.000 title claims abstract description 62
- 230000001072 progestational effect Effects 0.000 title claims abstract description 58
- 238000000034 method Methods 0.000 title claims abstract description 36
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims abstract description 43
- 238000001514 detection method Methods 0.000 claims abstract description 25
- 238000010811 Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry Methods 0.000 claims abstract description 19
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical class C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 claims abstract description 18
- 239000000583 progesterone congener Substances 0.000 claims abstract description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 60
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 30
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 22
- 239000012224 working solution Substances 0.000 claims description 21
- 239000010865 sewage Substances 0.000 claims description 19
- 239000000243 solution Substances 0.000 claims description 16
- 238000000605 extraction Methods 0.000 claims description 15
- 238000012360 testing method Methods 0.000 claims description 13
- 239000012071 phase Substances 0.000 claims description 12
- 238000001035 drying Methods 0.000 claims description 11
- 239000003480 eluent Substances 0.000 claims description 11
- 238000001914 filtration Methods 0.000 claims description 11
- 229910052757 nitrogen Inorganic materials 0.000 claims description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 10
- 239000012498 ultrapure water Substances 0.000 claims description 10
- 150000001875 compounds Chemical class 0.000 claims description 9
- 239000011521 glass Substances 0.000 claims description 9
- WWYNJERNGUHSAO-XUDSTZEESA-N (+)-Norgestrel Chemical compound O=C1CC[C@@H]2[C@H]3CC[C@](CC)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 WWYNJERNGUHSAO-XUDSTZEESA-N 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 8
- 238000004949 mass spectrometry Methods 0.000 claims description 7
- 229940053934 norethindrone Drugs 0.000 claims description 6
- VIKNJXKGJWUCNN-XGXHKTLJSA-N norethisterone Chemical compound O=C1CC[C@@H]2[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 VIKNJXKGJWUCNN-XGXHKTLJSA-N 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 239000000186 progesterone Substances 0.000 claims description 6
- 229960003387 progesterone Drugs 0.000 claims description 6
- RJKFOVLPORLFTN-PQIPVKAESA-N (8s,9s,10r,13s,14s,17s)-2,2,4,6,6,17-hexadeuterio-10,13-dimethyl-17-(2,2,2-trideuterioacetyl)-7,8,9,11,12,14,15,16-octahydro-1h-cyclopenta[a]phenanthren-3-one Chemical compound C([C@]1(C)[C@@]([2H])(C(=O)C([2H])([2H])[2H])CC[C@H]1[C@@H]1CC2([2H])[2H])C[C@@H]1[C@]1(C)C2=C([2H])C(=O)C([2H])([2H])C1 RJKFOVLPORLFTN-PQIPVKAESA-N 0.000 claims description 5
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical class [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 229960003996 chlormadinone Drugs 0.000 claims description 5
- VUHJZBBCZGVNDZ-TTYLFXKOSA-N chlormadinone Chemical compound C1=C(Cl)C2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@@](C(=O)C)(O)[C@@]1(C)CC2 VUHJZBBCZGVNDZ-TTYLFXKOSA-N 0.000 claims description 5
- 238000004807 desolvation Methods 0.000 claims description 5
- METQSPRSQINEEU-UHFFFAOYSA-N dihydrospirorenone Natural products CC12CCC(C3(CCC(=O)C=C3C3CC33)C)C3C1C1CC1C21CCC(=O)O1 METQSPRSQINEEU-UHFFFAOYSA-N 0.000 claims description 5
- 229960004845 drospirenone Drugs 0.000 claims description 5
- METQSPRSQINEEU-HXCATZOESA-N drospirenone Chemical compound C([C@]12[C@H]3C[C@H]3[C@H]3[C@H]4[C@@H]([C@]5(CCC(=O)C=C5[C@@H]5C[C@@H]54)C)CC[C@@]31C)CC(=O)O2 METQSPRSQINEEU-HXCATZOESA-N 0.000 claims description 5
- 229960004913 dydrogesterone Drugs 0.000 claims description 5
- JGMOKGBVKVMRFX-HQZYFCCVSA-N dydrogesterone Chemical compound C1=CC2=CC(=O)CC[C@@]2(C)[C@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 JGMOKGBVKVMRFX-HQZYFCCVSA-N 0.000 claims description 5
- 238000000132 electrospray ionisation Methods 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 5
- 239000007789 gas Substances 0.000 claims description 5
- 229960002985 medroxyprogesterone acetate Drugs 0.000 claims description 5
- PSGAAPLEWMOORI-PEINSRQWSA-N medroxyprogesterone acetate Chemical compound C([C@@]12C)CC(=O)C=C1[C@@H](C)C[C@@H]1[C@@H]2CC[C@]2(C)[C@@](OC(C)=O)(C(C)=O)CC[C@H]21 PSGAAPLEWMOORI-PEINSRQWSA-N 0.000 claims description 5
- 238000005374 membrane filtration Methods 0.000 claims description 5
- VKHAHZOOUSRJNA-GCNJZUOMSA-N mifepristone Chemical compound C1([C@@H]2C3=C4CCC(=O)C=C4CC[C@H]3[C@@H]3CC[C@@]([C@]3(C2)C)(O)C#CC)=CC=C(N(C)C)C=C1 VKHAHZOOUSRJNA-GCNJZUOMSA-N 0.000 claims description 5
- 229960003248 mifepristone Drugs 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 239000011148 porous material Substances 0.000 claims description 5
- 238000002203 pretreatment Methods 0.000 claims description 5
- 230000004044 response Effects 0.000 claims description 5
- 235000011121 sodium hydroxide Nutrition 0.000 claims description 5
- 239000007790 solid phase Substances 0.000 claims description 5
- 239000012086 standard solution Substances 0.000 claims description 5
- 239000011550 stock solution Substances 0.000 claims description 5
- 238000004704 ultra performance liquid chromatography Methods 0.000 claims description 5
- 238000001291 vacuum drying Methods 0.000 claims description 5
- 238000010926 purge Methods 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 2
- 239000003463 adsorbent Substances 0.000 claims 1
- 150000001345 alkine derivatives Chemical class 0.000 claims 1
- 238000004458 analytical method Methods 0.000 abstract description 13
- 230000000694 effects Effects 0.000 abstract description 9
- 230000007613 environmental effect Effects 0.000 abstract description 8
- 239000011159 matrix material Substances 0.000 abstract description 8
- 239000000126 substance Substances 0.000 abstract description 7
- 238000004811 liquid chromatography Methods 0.000 abstract 1
- 238000001179 sorption measurement Methods 0.000 description 12
- 238000011084 recovery Methods 0.000 description 10
- VBOBXELNVGBCBR-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenylpyrrolidin-2-one Chemical compound C=CN1CCCC1=O.C=CC1=CC=CC=C1C=C VBOBXELNVGBCBR-UHFFFAOYSA-N 0.000 description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 8
- 206010067868 Skin mass Diseases 0.000 description 8
- 239000002131 composite material Substances 0.000 description 8
- 239000008103 glucose Substances 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 8
- 239000011347 resin Substances 0.000 description 8
- 229920005989 resin Polymers 0.000 description 8
- 239000000741 silica gel Substances 0.000 description 8
- 229910002027 silica gel Inorganic materials 0.000 description 8
- 229960001866 silicon dioxide Drugs 0.000 description 8
- 239000003795 chemical substances by application Substances 0.000 description 7
- 239000004519 grease Substances 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 4
- 241000208340 Araliaceae Species 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 4
- 235000003140 Panax quinquefolius Nutrition 0.000 description 4
- 235000019253 formic acid Nutrition 0.000 description 4
- 235000008434 ginseng Nutrition 0.000 description 4
- 125000005456 glyceride group Chemical group 0.000 description 4
- 229960004400 levonorgestrel Drugs 0.000 description 4
- 239000000314 lubricant Substances 0.000 description 4
- 238000001556 precipitation Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 238000004885 tandem mass spectrometry Methods 0.000 description 4
- 239000012535 impurity Substances 0.000 description 3
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 3
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 2
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 2
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 2
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 2
- 239000005642 Oleic acid Substances 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000002470 solid-phase micro-extraction Methods 0.000 description 2
- PHYFQTYBJUILEZ-IUPFWZBJSA-N triolein Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(OC(=O)CCCCCCC\C=C/CCCCCCCC)COC(=O)CCCCCCC\C=C/CCCCCCCC PHYFQTYBJUILEZ-IUPFWZBJSA-N 0.000 description 2
- 241000036318 Callitris preissii Species 0.000 description 1
- 206010058314 Dysplasia Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000031320 Teratogenesis Diseases 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 230000003749 cleanliness Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000032678 sex differentiation Effects 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/08—Preparation using an enricher
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/045—Standards internal
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses method that is a kind of while detecting a variety of progestational hormone in environment water,Belong to Analysis of Organic Substances field in environment,Pre-processed including water sample,New solid phase extraction adsorbentses Oasis PRiME HLB enrichments concentration,UPLC MS/MS determine progestin concentration,The drafting of standard curve and upper machine determine five steps,Using preconditioning techniques of the new solid phase extraction adsorbentses Oasis PRiME HLB as environmental water sample,Both the selectivity to determinand had been improved,Also enrichment times are considerably increased and reduce matrix effect,And instrument is used in conjunction as detection quantitative tool using superelevation liquid chromatography mass spectrometric,Determine 11 kinds of different progestational hormone in water environment simultaneously,Compared with common detection mode,Realize the high selectivity to progestational hormone and highly sensitive effectively combination,It is and fast with analyze speed,The advantages that use range is wide.
Description
Technical field
The invention belongs to Analysis of Organic Substances technical field in environment water, and in particular to a kind of to detect simultaneously in environment water
The method of a variety of progestational hormone.
Background technology
In half a century in past, the natural environment that progestational hormone is depended on for existence to the mankind and wild animal causes serious danger
Evil, have become common problem of environmental pollution urgently to be resolved hurrily in global range, cause the extensive concern of people.Progestational hormone is made
Environment can be accumulated in for a long time for a kind of common incretion interferent and in human body, metabolism is slow, can pass through diffusion, convective transfer
To air, soil, in the various surrounding mediums such as waters deposit.Once into body, it is normally interior can not only to upset body
The synthesis and metabolism of secreting hormone, influence is grown and Sex Differentiation, moreover it is possible to causes the multisystem such as nerve, endocrine, immune
Dysplasia and dysgenesia, or even teratogenesis and carcinogenic etc..Therefore, establish in quick, efficient, accurate, sensitive environment water
Progestational hormone analyzing detecting method is extremely important.
Due to environmental sample form of diverse, complicated component and progestogen content are relatively low the characteristics of, it is difficult to directly it is entered
Row analysis detection.In order to reduce the interference of sample substrate and improve analysis detection sensitivity, must enter before sample analysis is carried out
The suitable sample pretreatment of row could carry out various Instrumental Analysis.The conventional method of environmental hormone preconditioning technique in environmental sample
There are ultrasonic wave extraction, surname extraction, microwave abstracting etc..But ultrasonic wave extraction solvent-oil ratio is big, surname extraction time length, and needs
Want reflux operation；Need to filter after microwave abstracting, it is difficult to instrument on―line.At present, with the development of science and technology, many new samples
Product preconditioning technique arises at the historic moment, such as SPE, SPME.SPME technologies are as a kind of new Sample Pretreatment Technique, as it should
With the continuous expansion in field, but some limitations of its extracting fiber in itself cause new problem to continuously emerge, the rate of recovery and repetition
Property cannot be guaranteed.And a kind of new technique that SPE can pre-process as environmental hormone, its rate of recovery is higher and reappearance compared with
It is stable, but due to environmental sample form of diverse, complicated component and determinand content are relatively low the characteristics of, SPE technologies are proposed
Higher requirement.Oasis PRiME HLB can simplify and speed up SPE streams as a kind of new anti-phase solid phase extraction adsorbentses
Journey, it can provide cleaner sample within the shorter time, make the analysis of liquid chromatogram and LC-MS system more light
Pine.Oasis PRiME HLB posts can also can simplify extraction procedure with porous plate and accelerate rate of extraction, it is ensured that obtain it is consistent,
Reproducible liquid phase and LC-MS analysis result.Sample is prepared compared to using other abstraction techniques, assay laboratory is present
It can handle sample more quickly, speed lifting highest 40%, and the raising of sample cleanliness factor is up to 70%, and LC-MS
Matrix effect it is smaller.More other sample-pretreating methods can obtain cleaner extract simultaneously.Oasis PRiME HLB
It is a extremely successful anti-phase SPE products of new generation, it can be obtained more by simpler scheme, shorter processing time
Clean extract, the progestational hormone that can be effectively enriched with concentration environment water, it can be used as pregnant in a kind of processing environment water body
The preprocess method of hormone.
The content of the invention
For above technical problem, the present invention provides method that is a kind of while detecting a variety of progestational hormone in environment water, energy
It is effective to remove environment water impurity, greatly reduce matrix effect.
The technical scheme is that：Method that is a kind of while detecting a variety of progestational hormone in environment water, by testing sample
Detected after new solid phase extraction adsorbentses Oasis PRiME HLB pretreatments, then with UPLC-MS/MS, including following step
Suddenly：
(1) water sample pre-processes
The water sample of collection is subjected to three-stage filtration, the water sample after accurate measuring filtering using the filter membrane of three kinds of different pore sizes
0.2-2L, and be that 0.1mol/L hydrochloric acid and 0.1mol/L sodium hydroxides adjust pH to 7.0-7.2 using concentration；
(2) new solid phase extraction adsorbentses Oasis PRiME HLB enrichments concentration
The direct Oasis PRiME HLB solid phase extraction columns of crossing of water sample that step (1) is treated are enriched with, flow velocity control
Make in 7-9mL/min, the solid phase extraction column is eluted at twice using ultra-pure water after enrichment terminates, then by after elution
Solid phase extraction column dries 1-3h in vacuum drying chamber, and drying temperature is 25-35 DEG C, using ethyl acetate to dried solid
Mutually extraction pillar is eluted three times, obtains the eluent containing target compound, and the eluent is moved in nitrogen drying instrument,
The nitrogen for being 1.5-2m/s with flow velocity purges 30-60min, and it is standby to obtain residue；
(3) UPLC-MS/MS determines progestin concentration
Preparing standard solution：Accurately weigh the standard items that 1mg detects progestational hormone respectively, and respectively using methanol constant volume in
10mg/L mixed mark stock solution is obtained in 100mL volumetric flask, after mixing, is stored in -20 DEG C of refrigerators；
Prepare working solution：Precision measures the residue in the μ L dissolving steps (2) of methanol solution 500, after vortex, uses glass
Glass dropper constant volume 1mL is simultaneously diluted with 0.22 μm of membrane filtration, again with methanol solution, obtains the working solution of various concentrations；
Chromatographic condition：Chromatographic column ACQUITY UPLC BEH C18,2.1 × 100mm, 1.7 μm；Mobile phase A is 0.1%
Formic acid, Mobile phase B is methanol, using gradient elution program；Flow velocity is 0.3mL/min；Column temperature is 40 DEG C, and sample size is 10 μ L；
Wherein described gradient elution program is as follows：
| Time (min) | A (%) | B (%) |
| 0 | 10 | 90 |
| 0.5 | 60 | 40 |
| 3 | 65 | 35 |
| 6 | 70 | 30 |
| 6.5 | 100 | 0 |
| 8 | 10 | 90 |
Mass Spectrometry Conditions：Using more reaction detection patterns, ion gun is electro-spray ionization source (ESI sources), source temperature 120
DEG C, taper hole voltage：30V, extraction ion voltage are 2.5kV, and capillary voltage 3kV, desolvation temperature is 450 DEG C, precipitation
Agent gas velocity is 900L/hr；Sweep time is 0.1s；
(4) drafting of standard curve
To mix mark stock concentrations (mg/L) described in step (3) as abscissa, response, which is that ordinate drafting is pregnant, to swash
The standard curve of element；
(5) machine determines on
Working solution described in step (3) is detected using UPLC-MS/MS instruments, obtains the typical color of actual sample
Spectrogram, carry out qualitative detection is compared with the standard curve for the actual sample typical case chromatogram；Reuse UPLC-
The tuber function of MS/MS instruments, parent ion, daughter ion, collision voltage and the capillary voltage of each material are determined, uses above-mentioned ginseng
Several progestational hormone in working solution carry out quantitative detection.
Further, the progestational hormone is respectively mifepristone, chlormadinone, medroxyproges-terone acetate, Drospirenone, progesterone, first
Norethindrone, Dydrogesterone, Levonorgestrel, norethindrone, progesterone-d9, norethindrone-d6.
Further, the environment water is sewage treatment plant inflow, water outlet and river, and two kinds of water source human contacts lead
Height, it is representative stronger.
Further, when the water sample is sewage treatment plant inflow water sample, collection capacity is 200- in the step (1)
500mL, when water sample is sewage disposal plant effluent water sample, collection capacity is 300-500mL, and when water sample is river, collection capacity is 1-
2L。
Further, filled out inside Oasis PRiME HLB solid phase extraction columns described in step (2) containing composite adsorption
Material, the composite adsorption stuffing is that mass ratio is 2:1:1 poly N-vinyl pyrrolidone-divinylbenzene macroreticular resin, three oil
Acid glyceride fibrous nodules, hud typed glucose polymer-silica-gel sphere, the poly N-vinyl pyrrolidone-divinylbenzene macropore
Resin provides good water logging lubricant nature by one " special polarity captures group " come the reservation increased to polar substances；It is described
Olein fibrous nodules have stronger lipophilicity, to this quasi-grease compound strong adsorption force of progestational hormone and performance it is steady
It is fixed；Hud typed glucose polymer-the silica-gel sphere has highly hydrophilic, and concentration and separation works well.
Further, the Oasis PRiME HLB solid phase extraction columns do not have to be activated and balanced, and greatly save
Operating time and solvent.
Further, the aperture of filter membrane is respectively 10 μm, 1 μm and 0.45 μm described in step (1), and gradient filtration can be most
Bigization reduces impurity effect analysis result.
Further, the dosage of ultra-pure water and ethyl acetate described in step (2) is the solid phase extraction column volume
1-2 times.
Compared with prior art, beneficial effects of the present invention are：
(1) present invention is using pretreatment skills of the new solid phase extraction adsorbentses Oasis PRiME HLB as environmental water sample
Art, environment water impurity can be effectively removed, greatly reduces matrix effect.And UPLC-MS/MS is combined as detector, energy
The detection environment water progestogen content of enough fast accurates, testing result are accurate, reliable.
(2) present invention had both improved the selection to determinand using new solid phase extraction adsorbentses Oasis PRiME HLB
Property, also considerably increase enrichment times and reduce matrix effect, new approaches are provided for the development and application of pretreatment technology,
For the extraction to Environmental Trace medicament residue, concentrate and add new method with separating.
(3) instrument is used in conjunction to detect quantitative tool using superelevation liquid phase-mass spectrum in the present invention, while determines in water environment 11 kinds not
Same progestational hormone, compared with common detection mode, the high selectivity to progestational hormone and highly sensitive effectively combination are realized, and
Have the advantages that analyze speed is fast, use range is wide.
Brief description of the drawings
Fig. 1 is influence of the solid phase extraction column species of embodiment 2 to 9 kinds of progestational hormone rate of recovery；
Fig. 2 is influences of the different pH of embodiment 2 to 9 kinds of progestational hormone rate of recovery；
Fig. 3 is influence of the different leacheates of embodiment 2 to 9 kinds of progestational hormone；
Fig. 4 is influence of the different eluents of embodiment 2 to 9 kinds of progestational hormone rate of recovery；
Fig. 5 .1-5.3 are that 11 kinds of progestational hormone of embodiment 2 extract ion flow graph.
Embodiment
Embodiment 1
Method that is a kind of while detecting a variety of progestational hormone in environment water, wherein, the progestational hormone be respectively mifepristone,
Chlormadinone, medroxyproges-terone acetate, Drospirenone, progesterone, dl-Norgestrel, Dydrogesterone, Levonorgestrel, norethindrone, progesterone-d9,
Norethindrone-d6.By testing sample after new solid phase extraction adsorbentses Oasis PRiME HLB pretreatments, then use UPLC-
MS/MS is detected, and is comprised the following steps：
(1) water sample pre-processes
It is water sample to choose sewage treatment plant inflow, water outlet and river, and two kinds of water source human contacts lead height, representative stronger.
The water sample of collection is subjected to three-stage filtration using the filter membrane of 10 μm, 1 μm and 0.45 μm three kinds of different pore sizes, after accurate measuring filtering
Water sample, wherein, sewage treatment plant inflow water sample is 200mL, and sewage disposal plant effluent water sample is 300mL, and river collection capacity is
1L, and be that 0.1mol/L hydrochloric acid and 0.1mol/L sodium hydroxides adjust pH to 7.0 using concentration；
(2) new solid phase extraction adsorbentses Oasis PRiME HLB enrichments concentration
Oasis PRiME HLB solid phase extraction columns do not have to be activated and balanced, and greatly save operating time and molten
Agent, the direct Oasis PRiME HLB solid phase extraction columns of crossing of water sample that step (1) is treated are enriched with, and flow control exists
7mL/min, the solid phase extraction column is eluted at twice using ultra-pure water after enrichment terminates, then extract the solid phase after elution
Pillar is taken to dry 1h in vacuum drying chamber, drying temperature is 25 DEG C, using ethyl acetate to dried solid phase extraction column
Eluted three times, wherein, the dosage of the ultra-pure water and ethyl acetate is 1 times of the solid phase extraction column volume, is obtained
To the eluent containing target compound, the eluent is moved in nitrogen drying instrument, the nitrogen for being 1.5m/s with flow velocity purges
30min, it is standby to obtain residue；Wherein, composite adsorption stuffing is contained inside the Oasis PRiME HLB solid phase extraction columns,
The composite adsorption stuffing is that mass ratio is 2:1:1 poly N-vinyl pyrrolidone-divinylbenzene macroreticular resin, three oleic acid are sweet
Grease fibrous nodules, hud typed glucose polymer-silica-gel sphere, the poly N-vinyl pyrrolidone-divinylbenzene macroreticular resin
By one " special polarity captures group " good water logging lubricant nature is provided come the reservation increased to polar substances；Three oil
Acid glyceride fibrous nodules have stronger lipophilicity, to this quasi-grease compound strong adsorption force of progestational hormone and stable performance；Institute
Stating hud typed glucose polymer-silica-gel sphere has highly hydrophilic, and concentration and separation works well.
(3) UPLC-MS/MS determines progestin concentration
Preparing standard solution：Accurately weigh the standard items that 1mg detects progestational hormone respectively, and respectively using methanol constant volume in
10mg/L mixed mark stock solution is obtained in 100mL volumetric flask, after mixing, is stored in -20 DEG C of refrigerators；
Prepare working solution：Precision measures the residue in the μ L dissolving steps (2) of methanol solution 500, after vortex, uses glass
Glass dropper constant volume 1mL is simultaneously diluted with 0.22 μm of membrane filtration, again with methanol solution, obtains the working solution of various concentrations；
Chromatographic condition：Chromatographic column ACQUITY UPLC BEH C18,2.1 × 100mm, 1.7 μm；Mobile phase A is 0.1%
Formic acid, Mobile phase B is methanol, using gradient elution program；Flow velocity is 0.3mL/min；Column temperature is 40 DEG C, and sample size is 10 μ L；
Wherein described gradient elution program is as follows：
| Time (min) | A (%) | B (%) |
| 0 | 10 | 90 |
| 0.5 | 60 | 40 |
| 3 | 65 | 35 |
| 6 | 70 | 30 |
| 6.5 | 100 | 0 |
| 8 | 10 | 90 |
Mass Spectrometry Conditions：Using more reaction detection patterns, ion gun is electro-spray ionization source (ESI sources), source temperature 120
DEG C, taper hole voltage：30V, extraction ion voltage are 2.5kV, and capillary voltage 3kV, desolvation temperature is 450 DEG C, precipitation
Agent gas velocity is 900L/hr；Sweep time is 0.1s；
(4) drafting of standard curve
To mix mark stock concentrations (mg/L) described in step (3) as abscissa, response, which is that ordinate drafting is pregnant, to swash
The standard curve of element；
(5) machine determines on
Working solution described in step (3) is detected using UPLC-MS/MS instruments, obtains the typical color of actual sample
Spectrogram, carry out qualitative detection is compared with the standard curve for the actual sample typical case chromatogram；Reuse UPLC-
The tuber function of MS/MS instruments, parent ion, daughter ion, collision voltage and the capillary voltage of each material are determined, uses above-mentioned ginseng
Several progestational hormone in working solution carry out quantitative detection.
Embodiment 2
Method that is a kind of while detecting a variety of progestational hormone in environment water, wherein, the progestational hormone be respectively mifepristone,
Chlormadinone, medroxyproges-terone acetate, Drospirenone, progesterone, dl-Norgestrel, Dydrogesterone, Levonorgestrel, norethindrone, progesterone-d9,
Norethindrone-d6.By testing sample after new solid phase extraction adsorbentses Oasis PRiME HLB pretreatments, then use UPLC-
MS/MS is detected, and is comprised the following steps：
(1) water sample pre-processes
It is water sample to choose sewage treatment plant inflow, water outlet and river, and two kinds of water source human contacts lead height, representative stronger.
The water sample of collection is subjected to three-stage filtration using the filter membrane of 10 μm, 1 μm and 0.45 μm three kinds of different pore sizes, after accurate measuring filtering
Water sample, wherein, sewage treatment plant inflow water sample is 350mL, and sewage disposal plant effluent water sample is 400mL, and river collection capacity is
1.5L, and be that 0.1mol/L hydrochloric acid and 0.1mol/L sodium hydroxides adjust pH to 7.1 using concentration；
(2) new solid phase extraction adsorbentses Oasis PRiME HLB enrichments concentration
Oasis PRiME HLB solid phase extraction columns do not have to be activated and balanced, and greatly save operating time and molten
Agent, the direct Oasis PRiME HLB solid phase extraction columns of crossing of water sample that step (1) is treated are enriched with, and flow control exists
8mL/min, the solid phase extraction column is eluted at twice using ultra-pure water after enrichment terminates, then extract the solid phase after elution
Pillar is taken to dry 2h in vacuum drying chamber, drying temperature is 30 DEG C, using ethyl acetate to dried solid phase extraction column
Eluted three times, wherein, the dosage of the ultra-pure water and ethyl acetate is 1.5 times of the solid phase extraction column volume,
The eluent containing target compound is obtained, the eluent is moved in nitrogen drying instrument, the nitrogen for being 1.8m/s with flow velocity blows
45min is swept, it is standby to obtain residue；Wherein, filled out inside the Oasis PRiME HLB solid phase extraction columns containing composite adsorption
Material, the composite adsorption stuffing is that mass ratio is 2:1:1 poly N-vinyl pyrrolidone-divinylbenzene macroreticular resin, three oil
Acid glyceride fibrous nodules, hud typed glucose polymer-silica-gel sphere, the poly N-vinyl pyrrolidone-divinylbenzene macropore
Resin provides good water logging lubricant nature by one " special polarity captures group " come the reservation increased to polar substances；It is described
Olein fibrous nodules have stronger lipophilicity, to this quasi-grease compound strong adsorption force of progestational hormone and performance it is steady
It is fixed；Hud typed glucose polymer-the silica-gel sphere has highly hydrophilic, and concentration and separation works well.
(3) UPLC-MS/MS determines progestin concentration
Preparing standard solution：Accurately weigh the standard items that 1mg detects progestational hormone respectively, and respectively using methanol constant volume in
10mg/L mixed mark stock solution is obtained in 100mL volumetric flask, after mixing, is stored in -20 DEG C of refrigerators；
Prepare working solution：Precision measures the residue in the μ L dissolving steps (2) of methanol solution 500, after vortex, uses glass
Glass dropper constant volume 1mL is simultaneously diluted with 0.22 μm of membrane filtration, again with methanol solution, obtains the working solution of various concentrations；
Chromatographic condition：Chromatographic column ACQUITY UPLC BEH C18,2.1 × 100mm, 1.7 μm；Mobile phase A is 0.1%
Formic acid, Mobile phase B is methanol, using gradient elution program；Flow velocity is 0.3mL/min；Column temperature is 40 DEG C, and sample size is 10 μ L；
Wherein described gradient elution program is as follows：
| Time (min) | A (%) | B (%) |
| 0 | 10 | 90 |
| 0.5 | 60 | 40 |
| 3 | 65 | 35 |
| 6 | 70 | 30 |
| 6.5 | 100 | 0 |
| 8 | 10 | 90 |
Mass Spectrometry Conditions：Using more reaction detection patterns, ion gun is electro-spray ionization source (ESI sources), source temperature 120
DEG C, taper hole voltage：30V, extraction ion voltage are 2.5kV, and capillary voltage 3kV, desolvation temperature is 450 DEG C, precipitation
Agent gas velocity is 900L/hr；Sweep time is 0.1s；
(4) drafting of standard curve
To mix mark stock concentrations (mg/L) described in step (3) as abscissa, response, which is that ordinate drafting is pregnant, to swash
The standard curve of element；
(5) machine determines on
Working solution described in step (3) is detected using UPLC-MS/MS instruments, obtains the typical color of actual sample
Spectrogram, carry out qualitative detection is compared with the standard curve for the actual sample typical case chromatogram；Reuse UPLC-
The tuber function of MS/MS instruments, parent ion, daughter ion, collision voltage and the capillary voltage of each material are determined, uses above-mentioned ginseng
Several progestational hormone in working solution carry out quantitative detection.
Embodiment 3
Method that is a kind of while detecting a variety of progestational hormone in environment water, wherein, the progestational hormone be respectively mifepristone,
Chlormadinone, medroxyproges-terone acetate, Drospirenone, progesterone, dl-Norgestrel, Dydrogesterone, Levonorgestrel, norethindrone, progesterone-d9,
Norethindrone-d6.By testing sample after new solid phase extraction adsorbentses Oasis PRiME HLB pretreatments, then use UPLC-
MS/MS is detected, and is comprised the following steps：
(1) water sample pre-processes
It is water sample to choose sewage treatment plant inflow, water outlet and river, and two kinds of water source human contacts lead height, representative stronger.
The water sample of collection is subjected to three-stage filtration using the filter membrane of 10 μm, 1 μm and 0.45 μm three kinds of different pore sizes, after accurate measuring filtering
Water sample, wherein, sewage treatment plant inflow water sample is 500mL, and sewage disposal plant effluent water sample is 500mL, and river collection capacity is
2L, and be that 0.1mol/L hydrochloric acid and 0.1mol/L sodium hydroxides adjust pH to 7.2 using concentration；
(2) new solid phase extraction adsorbentses Oasis PRiME HLB enrichments concentration
Oasis PRiME HLB solid phase extraction columns do not have to be activated and balanced, and greatly save operating time and molten
Agent, the direct Oasis PRiME HLB solid phase extraction columns of crossing of water sample that step (1) is treated are enriched with, and flow control exists
9mL/min, the solid phase extraction column is eluted at twice using ultra-pure water after enrichment terminates, then extract the solid phase after elution
Pillar is taken to dry 3h in vacuum drying chamber, drying temperature is 35 DEG C, using ethyl acetate to dried solid phase extraction column
Eluted three times, wherein, the dosage of the ultra-pure water and ethyl acetate is 2 times of the solid phase extraction column volume, is obtained
To the eluent containing target compound, the eluent is moved in nitrogen drying instrument, the nitrogen for being 2m/s with flow velocity purges
60min, it is standby to obtain residue；Wherein, composite adsorption stuffing is contained inside the Oasis PRiME HLB solid phase extraction columns,
The composite adsorption stuffing is that mass ratio is 2:1:1 poly N-vinyl pyrrolidone-divinylbenzene macroreticular resin, three oleic acid are sweet
Grease fibrous nodules, hud typed glucose polymer-silica-gel sphere, the poly N-vinyl pyrrolidone-divinylbenzene macroreticular resin
By one " special polarity captures group " good water logging lubricant nature is provided come the reservation increased to polar substances；Three oil
Acid glyceride fibrous nodules have stronger lipophilicity, to this quasi-grease compound strong adsorption force of progestational hormone and stable performance；Institute
Stating hud typed glucose polymer-silica-gel sphere has highly hydrophilic, and concentration and separation works well.
(3) UPLC-MS/MS determines progestin concentration
Preparing standard solution：Accurately weigh the standard items that 1mg detects progestational hormone respectively, and respectively using methanol constant volume in
10mg/L mixed mark stock solution is obtained in 100mL volumetric flask, after mixing, is stored in -20 DEG C of refrigerators；
Prepare working solution：Precision measures the residue in the μ L dissolving steps (2) of methanol solution 500, after vortex, uses glass
Glass dropper constant volume 1mL is simultaneously diluted with 0.22 μm of membrane filtration, again with methanol solution, obtains the working solution of various concentrations；
Chromatographic condition：Chromatographic column ACQUITY UPLC BEH C18,2.1 × 100mm, 1.7 μm；Mobile phase A is 0.1%
Formic acid, Mobile phase B is methanol, using gradient elution program；Flow velocity is 0.3mL/min；Column temperature is 40 DEG C, and sample size is 10 μ L；
Wherein described gradient elution program is as follows：
| Time (min) | A (%) | B (%) |
| 0 | 10 | 90 |
| 0.5 | 60 | 40 |
| 3 | 65 | 35 |
| 6 | 70 | 30 |
| 6.5 | 100 | 0 |
| 8 | 10 | 90 |
Mass Spectrometry Conditions：Using more reaction detection patterns, ion gun is electro-spray ionization source (ESI sources), source temperature 120
DEG C, taper hole voltage：30V, extraction ion voltage are 2.5kV, and capillary voltage 3kV, desolvation temperature is 450 DEG C, precipitation
Agent gas velocity is 900L/hr；Sweep time is 0.1s；
(4) drafting of standard curve
To mix mark stock concentrations (mg/L) described in step (3) as abscissa, response, which is that ordinate drafting is pregnant, to swash
The standard curve of element；
(5) machine determines on
Working solution described in step (3) is detected using UPLC-MS/MS instruments, obtains the typical color of actual sample
Spectrogram, carry out qualitative detection is compared with the standard curve for the actual sample typical case chromatogram；Reuse UPLC-
The tuber function of MS/MS instruments, parent ion, daughter ion, collision voltage and the capillary voltage of each material are determined, uses above-mentioned ginseng
Several progestational hormone in working solution carry out quantitative detection.
Interpretation of result
It is the upper machine measurement result analysis of embodiment 3 below：
(1) qualitative detection
Wherein, 0.1-0.8ng/L, high sensitivity are limited to 3 times of test limits for method of signal to noise ratio, then detection；And line
For property coefficient correlation more than 0.99, linear relationship is good.Standard curve range, coefficient correlation, the test limit of each working solution
It is as shown in table 1 etc. result：
The methodology testing result of table 1
(2) quantitative detection
Reuse the tuber function of UPLC-MS/MS instruments, determine the parent ion of each material, daughter ion, collision voltage and
Capillary voltage, quantitative detection is carried out to the progestational hormone in working solution using above-mentioned parameter.Wherein, 11 kinds of progestational hormone quantitative analyses
Mass spectrometry parameters are as shown in table 2：
The Mass Spectrometry Conditions of 2 11 kinds of progestational hormone of table
(3) matrix effect calculates
A：Internal standard and standard liquid are added in the water sample by above-mentioned steps (1) processing, according to step (2) to above-mentioned pregnant sharp
Element is extracted and detected according to step (3), is as a result defined as Cm；
B：Identical water sample, is not added with internal standard and standard liquid, and above-mentioned progestational hormone is extracted according to step (2), from carrying
Take and 100 μ L are taken in liquid, nitrogen drying, then add the mixed mark solution (50 μ g/L) of 100 μ L progestational hormone, obtained result is defined as
Cm+100；
C：The standard liquid of the mixed mark solution (50 μ g/L) of progestational hormone is analyzed according to step (3), is as a result defined as C100；Then
The result of calculation of matrix effect (ME%) is ME%=[(Cm+100-Cm)/C100] × 100%, as a result as shown in table 3：
The progestational hormone matrix effect (ME%) of table 3
(4) recovery of standard addition calculates
D：Sewage treatment plant inflow and water outlet and river water sample are taken, adds the standard liquid of certain density each determinand
In the water sample handled by above-mentioned steps (1), after above-mentioned steps (2) enrichment concentration, by above-mentioned steps (3) test sample, obtain corresponding
As a result it is defined as Ca；
E：By the standard liquid of certain density each determinand, without processing, accordingly tied by above-mentioned steps (3) test sample
Fruit Cb；Then the calculation formula of the rate of recovery (RE%) is：RE%=Ca/Cb× 100%
The present invention has investigated the rate of recovery of 3 concentration, and its result is as shown in table 4, and the rate of recovery of each test substance is higher, full
Foot analysis requires.
The rate of recovery of the 4 each determinand of table in sewage treatment plant's Inlet and outlet water and river
(5) content analysis
Sewage treatment plant inflow and water outlet and river water sample are taken, the inner mark solution for adding certain density each determinand is pressed
In the water sample of above-mentioned steps (1) processing, after above-mentioned steps (2) enrichment concentration, by above-mentioned steps (3) test sample, progestational hormone is obtained
Content (ng/L) such as table 5；
Progestogen content (ng/L) in the sewage treatment plant of table 5 and river
Finally it should be noted that：The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although
The present invention is described in detail with reference to the foregoing embodiments, it will be understood by those within the art that：It still may be used
To be modified to the technical scheme described in previous embodiment, or equivalent substitution is carried out to which part technical characteristic；And
These modifications are replaced, and the essence of appropriate technical solution is departed from the spirit and model of technical scheme of the embodiment of the present invention
Enclose.
Claims (8)
1. method that is a kind of while detecting a variety of progestational hormone in environment water, it is characterised in that by testing sample by new solid
Mutually after extraction adsorbent Oasis PRiME HLB pretreatments, then detected with UPLC-MS/MS, comprised the following steps：
(1) water sample pre-processes
The water sample of collection is subjected to three-stage filtration, the water sample 0.2- after accurate measuring filtering using the filter membrane of three kinds of different pore sizes
2L, and be that 0.1mol/L hydrochloric acid and 0.1mol/L sodium hydroxides adjust pH to 7.0-7.2 using concentration；
(2) new solid phase extraction adsorbentses Oasis PRiME HLB enrichments concentration
The direct Oasis PRiME HLB solid phase extraction columns of crossing of water sample that step (1) is treated are enriched with, and flow control exists
7-9mL/min, the solid phase extraction column is eluted at twice using ultra-pure water after enrichment terminates, then by the solid phase after elution
Extract pillar and 1-3h is dried in vacuum drying chamber, drying temperature is 25-35 DEG C, and dried solid phase is extracted using ethyl acetate
Take pillar to be eluted three times, obtain the eluent containing target compound, the eluent is moved in nitrogen drying instrument, with stream
Speed purges 30-60min for 1.5-2m/s nitrogen, and it is standby to obtain residue；
(3) UPLC-MS/MS determines progestin concentration
Preparing standard solution：Accurately weigh the standard items that 1mg detects progestational hormone respectively, and respectively using methanol constant volume in
10mg/L mixed mark stock solution is obtained in 100mL volumetric flask, after mixing, is stored in -20 DEG C of refrigerators；
Prepare working solution：Precision measures the residue in the μ L dissolving steps (2) of methanol solution 500, after vortex, uses glass drop
Pipe constant volume 1mL is simultaneously diluted with 0.22 μm of membrane filtration, again with methanol solution, obtains the working solution of various concentrations；
Chromatographic condition：Chromatographic column ACQUITY UPLC BEH C18,2.1 × 100mm, 1.7 μm；Mobile phase A is 0.1% first
Acid, Mobile phase B is methanol, using gradient elution program；Flow velocity is 0.3mL/min；Column temperature is 40 DEG C, and sample size is 10 μ L；Its
Described in gradient elution program it is as follows：
Mass Spectrometry Conditions：Using more reaction detection patterns, ion gun is electro-spray ionization source (ESI sources), and source temperature is 120 DEG C,
Taper hole voltage：30V, extraction ion voltage are 2.5kV, and capillary voltage 3kV, desolvation temperature is 450 DEG C, desolventizing gas
Flow velocity is 900L/hr；Sweep time is 0.1s；
(4) drafting of standard curve
To mix mark stock concentrations (mg/L) described in step (3) as abscissa, response is that ordinate draws progestational hormone
Standard curve；
(5) machine determines on
Working solution described in step (3) is detected using UPLC-MS/MS instruments, obtains the typical chromatogram of actual sample,
Carry out qualitative detection is compared with the standard curve for the actual sample typical case chromatogram；Reuse UPLC-MS/MS instrument
The tuber function of device, parent ion, daughter ion, collision voltage and the capillary voltage of each material are determined, using above-mentioned parameter to work
Make the progestational hormone in solution and carry out quantitative detection.
2. method that is as claimed in claim 1 a kind of while detecting a variety of progestational hormone in environment water, it is characterised in that described
Progestational hormone is respectively mifepristone, chlormadinone, medroxyproges-terone acetate, Drospirenone, progesterone, dl-Norgestrel, Dydrogesterone, left alkynes promise
Progesterone, norethindrone, progesterone-d9, norethindrone-d6.
3. method that is as claimed in claim 1 a kind of while detecting a variety of progestational hormone in environment water, it is characterised in that described
Environment water is sewage treatment plant inflow, water outlet and river.
4. method that is as claimed in claim 1 a kind of while detecting a variety of progestational hormone in environment water, it is characterised in that described
When the water sample is sewage farm water inlet water sample, collection capacity is 200-500mL in step (1), when water sample is sewage farm
Collection capacity is 300-500mL during water outlet water sample, and when water sample is river, collection capacity is 1-2L.
5. method that is as claimed in claim 1 a kind of while detecting a variety of progestational hormone in environment water, it is characterised in that described
Oasis PRiME HLB solid phase extraction columns do not have to be activated and balanced.
6. method that is as claimed in claim 1 a kind of while detecting a variety of progestational hormone in environment water, it is characterised in that step
(1) aperture of filter membrane described in is respectively 10 μm and 1 μm.
7. method that is as claimed in claim 1 a kind of while detecting a variety of progestational hormone in environment water, it is characterised in that step
(1) aperture of filter membrane described in is respectively 10 μm, 1 μm and 0.45 μm.
8. method that is as claimed in claim 1 a kind of while detecting a variety of progestational hormone in environment water, it is characterised in that step
(2) dosage of ultra-pure water described in and ethyl acetate is 1-2 times of the solid phase extraction column volume.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105334282A (en) * | 2015-10-27 | 2016-02-17 | 哈尔滨工业大学 | Co-detecting method for environmental estrogens in surface water body |
| CN107121518A (en) * | 2017-06-30 | 2017-09-01 | 华东理工大学 | A kind of method of phenols, estrogens and androgens incretion interferent in Sync enrichment detection drinking water |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105334282A (en) * | 2015-10-27 | 2016-02-17 | 哈尔滨工业大学 | Co-detecting method for environmental estrogens in surface water body |
| CN107121518A (en) * | 2017-06-30 | 2017-09-01 | 华东理工大学 | A kind of method of phenols, estrogens and androgens incretion interferent in Sync enrichment detection drinking water |
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| DR EMMA L. WILLIAMS: "LC-MS/MS measurement of serum steroids in the clinical laboratory", 《MASS SPECTROMETRY》 * |
| 谭丽超: "水环境中类固醇激素的污染特征及健康风险评价研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
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