CN108107123A - The method that liquid chromatogram-triple quadrupole rods tandem mass spectrometries detect 7 kinds of artificial sweetening agents in environment water simultaneously - Google Patents
The method that liquid chromatogram-triple quadrupole rods tandem mass spectrometries detect 7 kinds of artificial sweetening agents in environment water simultaneously Download PDFInfo
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- 229910001868 water Inorganic materials 0.000 title claims abstract description 64
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 239000007788 liquid Substances 0.000 title claims abstract description 24
- 239000008122 artificial sweetener Substances 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 20
- 238000004885 tandem mass spectrometry Methods 0.000 title claims abstract description 18
- WEVYAHXRMPXWCK-UHFFFAOYSA-N acetonitrile Substances CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000010828 elution Methods 0.000 claims abstract description 14
- 238000004458 analytical method Methods 0.000 claims abstract description 13
- 239000012071 phase Substances 0.000 claims abstract description 13
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims abstract description 11
- 238000001514 detection method Methods 0.000 claims abstract description 10
- 238000010813 internal standard method Methods 0.000 claims abstract description 8
- 230000014759 maintenance of location Effects 0.000 claims abstract description 8
- WBZFUFAFFUEMEI-UHFFFAOYSA-M Acesulfame k Chemical compound [K+].CC1=CC(=O)[N-]S(=O)(=O)O1 WBZFUFAFFUEMEI-UHFFFAOYSA-M 0.000 claims abstract description 7
- 108010011485 Aspartame Proteins 0.000 claims abstract description 7
- 235000010358 acesulfame potassium Nutrition 0.000 claims abstract description 7
- 239000000619 acesulfame-K Substances 0.000 claims abstract description 7
- 239000000605 aspartame Substances 0.000 claims abstract description 7
- 235000010357 aspartame Nutrition 0.000 claims abstract description 7
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 claims abstract description 7
- 229960003438 aspartame Drugs 0.000 claims abstract description 7
- 235000012907 honey Nutrition 0.000 claims abstract description 7
- 239000004384 Neotame Substances 0.000 claims abstract description 6
- 238000005374 membrane filtration Methods 0.000 claims abstract description 6
- 235000019412 neotame Nutrition 0.000 claims abstract description 6
- HLIAVLHNDJUHFG-HOTGVXAUSA-N neotame Chemical compound CC(C)(C)CCN[C@@H](CC(O)=O)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 HLIAVLHNDJUHFG-HOTGVXAUSA-N 0.000 claims abstract description 6
- 108010070257 neotame Proteins 0.000 claims abstract description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 93
- 235000003599 food sweetener Nutrition 0.000 claims description 23
- 239000003765 sweetening agent Substances 0.000 claims description 23
- 239000011259 mixed solution Substances 0.000 claims description 19
- 230000007613 environmental effect Effects 0.000 claims description 17
- 239000000243 solution Substances 0.000 claims description 14
- 238000012545 processing Methods 0.000 claims description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 7
- 239000003643 water by type Substances 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 5
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 claims description 5
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- 235000019204 saccharin Nutrition 0.000 claims description 5
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 claims description 5
- 229940081974 saccharin Drugs 0.000 claims description 5
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 240000002853 Nelumbo nucifera Species 0.000 claims description 3
- 235000006508 Nelumbo nucifera Nutrition 0.000 claims description 3
- 235000006510 Nelumbo pentapetala Nutrition 0.000 claims description 3
- 240000000111 Saccharum officinarum Species 0.000 claims description 3
- 235000007201 Saccharum officinarum Nutrition 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- ITVGXXMINPYUHD-UHFFFAOYSA-N neohesperidin dihydrochalcone Chemical compound C1=C(O)C(OC)=CC=C1CCC(=O)C(C(=C1)O)=C(O)C=C1OC1C(OC2C(C(O)C(O)C(C)O2)O)C(O)C(O)C(CO)O1 ITVGXXMINPYUHD-UHFFFAOYSA-N 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 238000004454 trace mineral analysis Methods 0.000 abstract description 5
- 239000004376 Sucralose Substances 0.000 abstract description 4
- 235000019408 sucralose Nutrition 0.000 abstract description 4
- BAQAVOSOZGMPRM-QBMZZYIRSA-N sucralose Chemical compound O[C@@H]1[C@@H](O)[C@@H](Cl)[C@@H](CO)O[C@@H]1O[C@@]1(CCl)[C@@H](O)[C@H](O)[C@@H](CCl)O1 BAQAVOSOZGMPRM-QBMZZYIRSA-N 0.000 abstract description 4
- 239000012086 standard solution Substances 0.000 abstract description 3
- FTLYMKDSHNWQKD-UHFFFAOYSA-N (2,4,5-trichlorophenyl)boronic acid Chemical compound OB(O)C1=CC(Cl)=C(Cl)C=C1Cl FTLYMKDSHNWQKD-UHFFFAOYSA-N 0.000 abstract description 2
- 239000012452 mother liquor Substances 0.000 abstract description 2
- 229940085605 saccharin sodium Drugs 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 239000001329 FEMA 3811 Substances 0.000 abstract 1
- 229960004998 acesulfame potassium Drugs 0.000 abstract 1
- 238000004587 chromatography analysis Methods 0.000 abstract 1
- ITVGXXMINPYUHD-CUVHLRMHSA-N neohesperidin dihydrochalcone Chemical compound C1=C(O)C(OC)=CC=C1CCC(=O)C(C(=C1)O)=C(O)C=C1O[C@H]1[C@H](O[C@H]2[C@@H]([C@H](O)[C@@H](O)[C@H](C)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 ITVGXXMINPYUHD-CUVHLRMHSA-N 0.000 abstract 1
- 229940089953 neohesperidin dihydrochalcone Drugs 0.000 abstract 1
- 235000010434 neohesperidine DC Nutrition 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- 150000002500 ions Chemical class 0.000 description 9
- 239000007789 gas Substances 0.000 description 7
- 238000005457 optimization Methods 0.000 description 6
- 229910021642 ultra pure water Inorganic materials 0.000 description 5
- 239000012498 ultrapure water Substances 0.000 description 5
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 3
- 235000019257 ammonium acetate Nutrition 0.000 description 3
- 239000003651 drinking water Substances 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 235000020188 drinking water Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000010865 sewage Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 101100348341 Caenorhabditis elegans gas-1 gene Proteins 0.000 description 1
- 241001247197 Cephalocarida Species 0.000 description 1
- 101100447658 Mus musculus Gas1 gene Proteins 0.000 description 1
- 101100447665 Mus musculus Gas2 gene Proteins 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 238000000889 atomisation Methods 0.000 description 1
- QUQPHWDTPGMPEX-UTWYECKDSA-N aurantiamarin Natural products COc1ccc(cc1O)[C@H]1CC(=O)c2c(O)cc(O[C@@H]3O[C@H](CO[C@@H]4O[C@@H](C)[C@H](O)[C@@H](O)[C@H]4O)[C@@H](O)[C@H](O)[C@H]3O)cc2O1 QUQPHWDTPGMPEX-UTWYECKDSA-N 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 235000012206 bottled water Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- QGGZBXOADPVUPN-UHFFFAOYSA-N dihydrochalcone Chemical compound C=1C=CC=CC=1C(=O)CCC1=CC=CC=C1 QGGZBXOADPVUPN-UHFFFAOYSA-N 0.000 description 1
- PXLWOFBAEVGBOA-UHFFFAOYSA-N dihydrochalcone Natural products OC1C(O)C(O)C(CO)OC1C1=C(O)C=CC(C(=O)CC(O)C=2C=CC(O)=CC=2)=C1O PXLWOFBAEVGBOA-UHFFFAOYSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 231100001261 hazardous Toxicity 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000010841 municipal wastewater Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 125000000185 sucrose group Chemical group 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 230000003442 weekly effect Effects 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
- G01N2030/062—Preparation extracting sample from raw material
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- Physics & Mathematics (AREA)
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Pathology (AREA)
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Abstract
The invention belongs to water body chemical analysis technical fields, disclose the method that a kind of triple quadrupole rods tandem mass spectrometries of liquid chromatogram detect 7 kinds of artificial sweetening agents in environment water simultaneously, including：Seven kinds of acesulfame potassium, honey element, saccharin sodium, Sucralose, Aspartame, neotame, neohesperidin dihydrochalcone artificial sweetening agent standard items are configured to the mother liquor of 1mg/mL respectively, then the mixed standard solution for possessing concentration gradient is prepared, it is separately added into 5ng/mL internal standards, establish standard curve, using C18 chromatography post separations, with H2O+5mMNH4Ac+0.5mMTris-acetonitrile is as mobile phase, by the way of gradient elution, the component isolated is carried out with retention time and mass-to-charge ratio qualitative, is quantified with internal standard method.To environment water sample with 0.45 μm of membrane filtration, after with the method for Solid Phase Extraction carry out purifying concentration, be finally detected analysis with the triple quadrupole rods tandem mass spectrometries of liquid chromatogram.The present invention can carry out trace analysis to 7 kinds of artificial sweetening agents in environment water simultaneously, and detection limits low, high sensitivity, and stability is good.
Description
Technical field
The present invention relates to technical field, more particularly to a kind of liquid chromatogram-triple quadrupole rods tandem mass spectrometries detect ambient water simultaneously
The method of 7 kinds of artificial sweetening agents in body.
Background technology
Artificial sweetening agent is a kind of artificial synthesized or semi-synthetic sucrose substituent, is hardly metabolized in human body, i.e., in body
Interior generates a small amount of heat, has catered to the diet of modern consumer (especially diabetes patient, obese people, eurodonticus) and has needed
It asks, in addition its relatively low production cost, thus is widely used in food and beverage.In addition, artificial sweetening agent in drug and
Also there is addition in personal care articles (such as cream and lip gloss).
It is widely used by long-term, has there is research to show artificial sweetening agent generally existing in the environment.Wherein for example
Acesulfame-K ACE, honey element CYC, saccharin SAC, Sucralose SUC are detected in municipal wastewater and some environment waters
It arrives, the concentration of especially ACE and SUC in water environment is apparently higher than personal care articles ingredient and other artificial synthetic organic chemistries
The concentration of substance.Also some researches show that 90% CYC and SAC can be disposed off into after crossing sewage treatment plant, but physico
Property more stable ACE and SUC is learned by sewage treatment plant's treated concentration there is no very big change occurs, therefore final
Potable water treatment plants product water in, the concentration of ACE and SUC can also reach 7 μ g/L and 2.4 μ g/L.Although toxicologic study
Show in the range of management and control, the use of artificial sweetening agent does not endanger human body significantly, but their accidents in the environment
In the presence of sizable concern is still caused, especially for it is verified that having the SUC up to the half-life period of several years in water.
Meanwhile be documented, SUC may influence cladocera and the physiology and motor behavior of biped.
Therefore, evaluation artificial sweetening agent environmental behaviour and it is potentially hazardous before, we it is necessary to establish it is a kind of detection limit
Low, clever lightness is high, and the good trace analysis detection method of stability goes the content of artificial sweetening agent in accurate determination of the environment water body.
The analyzing detecting method of current many maturations is mainly focused in drink and food, and there is no actual to apply to ambient water
Body, while the taking a broad survey for artificial sweetening agent compared to other western countries, our country's data in this respect are also very
It is few.Therefore, the foundation of this method contribute to more comprehensively, more accurately in investigation water environment in China manually people sweetener there are feelings
Condition ensures the safe and reliable of drinking water.
The detection of current most of artificial sweetening agents, most is liquid chromatogram-uv detection method, but its sensitivity
Low, stability is poor.
The content of the invention
The present invention provides a kind of liquid chromatogram-triple quadrupole rods tandem mass spectrometries and detects 7 kinds of artificial sweetening agents in environment water simultaneously
Method, solve the technical issues of detection sensitivity of artificial sweetening agent in environment water in the prior art is low, and stability is poor.
In order to solve the above technical problems, the present invention provides a kind of liquid chromatogram-triple quadrupole rods tandem mass spectrometries to detect ring simultaneously
The method of 7 kinds of artificial sweetening agents in the water body of border, including：
Standard items processing：
The standard items of powdered sweetener are taken respectively：Honey element CYC, saccharin SAC, acesulfame-K ACE, Aspartame ASP, trichlorine sugarcane
Sugared SUC, neotame NEO and neohesperidin dihydrochalcone NHDC are configured to the standard reserving solution of seven kinds of 1.00mg/mL with methanol；
Seven kinds of standard reserving solutions are mixed and seven kinds of standard mixed solutions of 1.00 μ g/mL are configured to methanol；
The internal standard standard items of powdered sweetener are taken respectively：ACE-d4, ASP-d5, SUC-d6, CYC-d11 are with 1:1 methanol/
Water is configured to 1.00 μ g/mL, tetra- kinds of internal standard mixed solutions；
It is 0.1ng/mL, 0.5ng/mL, 1ng/mL that seven kinds of standard mixed solutions are configured to concentration gradient with methanol,
The standard curve of 5ng/mL, 10ng/mL, 50ng/mL, 100ng/mL, 200ng/mL；Wherein, described the seven of each concentration gradient
Equal tetra- kinds of internal standard mixed solutions containing 5ng/mL in kind of standard mixed solution, are examined for liquid chromatogram-triple quadrupole rods tandem mass spectrometries
Survey analysis；
It is separated using C18 chromatographic columns, with H2O+5mMNH4Ac+0.5mMTris (trishydroxymethylaminomethane)-acetonitrile is
Mobile phase, gradient elution is qualitative to each component progress with retention time and mass-to-charge ratio, and seven kinds of sweeteners are quantified with internal standard method,
Obtain the regression equation and related coefficient of each sweetener；
Environmental water sample processing to be measured：
50mL environmental water samples are taken through 0.45 μm of membrane filtration, and the PH of the environmental water sample after filtering is adjusted to 2 with dilute hydrochloric acid；
Purifying concentration environmental water sample is carried out with Solid Phase Extraction：
5mL methanol is used respectively, and the 5mL ultra-pure waters of PH=2 activate Oasis HLB solid phase extraction columns；With per second 1 drop
Speed loading；It is eluted and cleaned with 10mL ultra-pure waters (PH=2), vacuum drains 30min；It is eluted with 4mL methanol, the elution that will be obtained
Liquid is blown to 0.5mL in 45 DEG C of nitrogen；It is 25 with volume ratio:75 methanol/water solution constant volume is to 1mL；12000rpm centrifuges 10 points
Clock takes supernatant to carry out LC-MS/MS analyses；
It learns from else's experience the environment water body example of the environmental water sample processing step to be measured processing, is separated using C18 chromatographic columns, with
H2O+5mMNH4Ac+0.5mMTris (trishydroxymethylaminomethane)-acetonitrile be mobile phase, gradient elution, with retention time and matter
Lotus is more qualitative than being carried out for the component isolated, and the concentration of seven kinds of sweeteners is obtained with internal standard method.
The one or more technical solutions provided in the embodiment of the present application, have at least the following technical effects or advantages：
The liquid chromatogram provided in the embodiment of the present application-triple quadrupole rods tandem mass spectrometries detect in environment water 7 kinds manually simultaneously
The method of sweetener using liquid chromatogram-triple quadrupole rods tandem mass spectrometries, to sample purify dense using solid phase extraction techniques
Contracting, can carry out trace analysis, and detection limits low, high sensitivity, and stability is good, contribute to more accurate, more accurately monitor water quality,
Ensure the safety of drinking water.
Description of the drawings
Fig. 1 is chromatogram provided in an embodiment of the present invention.
Specific embodiment
The embodiment of the present application detects in environment water 7 kinds simultaneously by providing a kind of liquid chromatogram-triple quadrupole rods tandem mass spectrometries
The method of artificial sweetening agent, solves that the detection spirit lightness of artificial sweetening agent in environment water in the prior art is low, stability difference
Technical problem.
In order to better understand the above technical scheme, in conjunction with appended figures and specific embodiments to above-mentioned skill
Art scheme is described in detail, it should be understood that the specific features in the embodiment of the present invention and embodiment are to present techniques side
The detailed description of case rather than the restriction to technical scheme, in the case where there is no conflict, the embodiment of the present application and
Technical characteristic in embodiment can be mutually combined.
The method that a kind of liquid chromatogram-triple quadrupole rods tandem mass spectrometries detect 7 kinds of artificial sweetening agents in environment water simultaneously, bag
It includes：
Standard items processing：
The standard items of powdered sweetener are taken respectively：Honey element CYC, saccharin SAC, acesulfame-K ACE, Aspartame ASP, trichlorine sugarcane
Sugared SUC, neotame NEO and neohesperidin dihydrochalcone NHDC are configured to the standard reserving solution of seven kinds of 1.00mg/mL with methanol；
Seven kinds of standard reserving solutions are configured to seven kinds of standard mixed solutions of 1.00 μ g/mL with methanol；
The internal standard standard items of powdered sweetener are taken respectively：ACE-d4, ASP-d5, SUC-d6, CYC-d11 are with 1:1 methanol/
Water is configured to 1.00 μ g/mL, tetra- kinds of internal standard mixed solutions；
It is 0.1ng/mL, 0.5ng/mL, 1ng/mL that seven kinds of standard mixed solutions are configured to concentration gradient with methanol,
The standard curve of 5ng/mL, 10ng/mL, 50ng/mL, 100ng/mL, 200ng/mL；Wherein, described the seven of each concentration gradient
Equal tetra- kinds of internal standard mixed solutions containing 5ng/mL in kind of standard mixed solution, are examined for liquid chromatogram-triple quadrupole rods tandem mass spectrometries
Survey analysis；
It is separated using C18 chromatographic columns, with H2O+5mMNH4Ac+0.5mMTris (trishydroxymethylaminomethane)-acetonitrile is
Mobile phase, gradient elution is qualitative to each component progress with retention time and mass-to-charge ratio, and seven kinds of sweeteners are quantified with internal standard method,
Obtain the regression equation and related coefficient of each sweetener；
Environmental water sample processing to be measured：
50mL environmental water samples are taken through 0.45 μm of membrane filtration, and the PH of the environmental water sample after filtering is adjusted to 2 with dilute hydrochloric acid；
Purifying concentration environmental water sample is carried out with Solid Phase Extraction：
5mL methanol is used respectively, and the 5mL ultra-pure waters of PH=2 activate Oasis HLB solid phase extraction columns；With per second 1 drop
Speed loading；It is eluted and cleaned with 10mL ultra-pure waters (PH=2), vacuum drains 30min；It is eluted with 4mL methanol, the elution that will be obtained
Liquid is blown to 0.5mL in 45 DEG C of nitrogen；It is 25 with volume ratio:75 methanol/water solution constant volume is to 1mL；12000rpm centrifuges 10 points
Clock takes supernatant to carry out LC-MS/MS analyses；
It learns from else's experience the environment water body example of the environmental water sample processing step to be measured processing, is separated using C18 chromatographic columns, with
H2O+5mMNH4Ac+0.5mMTris (trishydroxymethylaminomethane)-acetonitrile be mobile phase, gradient elution, with retention time and matter
Lotus is more qualitative than being carried out for the component isolated, and the concentration of seven kinds of sweeteners is obtained with internal standard method.
It will be illustrated below by specific embodiment.
1. experimental section
1.1 reagents and instrument
Ultra performance liquid chromatography-triple quadrupole rods tandem mass spectrometries instrument, such as Thermo companies UPLC U3000 high performance liquid chromatographs
4500 mass spectrographs of series connection AB sciex API；Vortex oscillator (MX-F).
Methanol, acetonitrile are chromatographically pure, and ammonium acetate (is analyzed pure), ultra-pure water；
Artificial sweetening agent standard items：It is honey element, acesulfame-K, Aspartame (aspartame), Sucralose, neotame, saccharin, new
Aurantiamarin dihydrochalcone；
Internal standard standard items：ACE-d4, ASP-d5, SUC-d6, CYC-d11.
By above-mentioned standard product be configured to respectively seven kinds of standard mixed solutions of standard reserving solution and 1.00 μ g/mL of 1.00mg/mL in-
It is preserved in 20 DEG C of refrigerators.
By internal standard standard items ACE-d4, ASP-d5, SUC-d6, CYC-d11 with 1:1 methanol/water is configured in tetra- kinds of 1.00 μ g/mL
Mixed solution is marked, deposits in -20 DEG C.
Standard curve：It is 0.1ng/mL, 0.5ng/mL that seven kinds of standard mixed solutions are configured to concentration gradient with methanol,
The standard curve of 1ng/mL, 5ng/mL, 10ng/mL, 50ng/mL, 100ng/mL, 200ng/mL, while add in tetra- kinds of 5ng/mL
Internal standard mixed solution is tested and analyzed with liquid chromatogram-three-level quadruple bar tandem mass spectrum.
Instrument parameter condition：
Liquid-phase condition：
Chromatographic column：C18,150 × 4.6mm, 5 μm
Sample size：10.00μL
Mobile phase：A：H2O+5mMNH4Ac+0.5mMTris；B：Acetonitrile
Column temperature：40±5℃
Type of elution：Gradient elution is shown in Table 1
1 gradient elution of table
| Step | Time | Flow (mL/min) | %A | %B | Curve |
| 1 | 0 | 0.6 | 5 | 95 | 5 |
| 2 | 8 | 0.6 | 75 | 25 | 5 |
| 3 | 10 | 0.6 | 75 | 25 | 5 |
| 4 | 11 | 0.6 | 5 | 95 | 5 |
| 5 | 18 | 0.6 | 5 | 95 | 5 |
Mass Spectrometry Conditions：
Scan mode：Reaction of high order monitors (MRM)；ESI；
Scan mode：Negative ion mode
Ion source temperature：450 DEG C,
Ion source voltage：-3500V
CUR (Curtain Gas) gas curtain gas 40Psi
CAD (CoLLision Gas) collision gas 8Psi
GS1 (Ion Source Gas1) atomization gas 50Psi
GS2 (Ion Source Gas2) auxiliary gas 50Psi
Environmental water sample pre-treating method：
50mL water samples are taken, by 0.45 μm of membrane filtration, adjust PH=2 with dilute hydrochloric acid (analysis is pure) afterwards.With 5mL methanol and 5mL
(PH=2) ultra-pure water activation Oasis HLB solid phase extraction columns, the speed loading dripped with per second 1, after sample flow is complete,
It is cleaned with the ultrapure water wash of 10mL PH=2, vacuum drains 30min, is finally eluted with the pure methanol of 4mL, the eluent that will be obtained
Nitrogen is blown to 0.5mL under conditions of 45 DEG C, uses MeOH:H2O=25:75(V：V) constant volume centrifuges 10 to 1mL, last 12000rpm
Minute, supernatant is taken to carry out LC-MS/MS analyses.
2. the optimization of testing conditions
The optimization of 2.1 chromatographic conditions
Common mobile phase methanol-water and acetonitrile-water are compared, the experimental results showed that when making mobile phase using acetonitrile-water
Separating degree is good, high sensitivity.Attempt supplement Mobile Phase Additives simultaneously, finally herein using 5mM ammonium acetates and 0.5mMTris as
Mobile Phase Additives, experiment show that the separating degree for optimizing later 7 kinds of sweeteners of condition is good, and response is high, and peak type is graceful, reaches
Trace analysis requirement.
The optimization of 2.2 Mass Spectrometry Conditions
Herein using ESI negative ion modes, multistage monitoring scan pattern prepares the single standard solution of 7 kinds of sweeteners respectively, will
High concentration mother liquor is diluted to 20ng/mL step by step with methanol, for the optimization of Mass Spectrometry Conditions, because going cluster voltage (DP) and impact energy
(CE) it is affected to the response of compound, therefore mainly optimizes DP and CE in mass spectrum optimization process, while finely tunes EP (entrances
Voltage), the response of each compound is made to reach optimum state.
2 parent ion of table, daughter ion, DP, EP, CE, CXP
Wherein：Asterisk (*) is quota ion.
Linear relationship：Sweetener titer 0.1ng/mL, 0.5ng/mL, 1ng/mL, the 5ng/mL of various concentration gradient are configured,
10ng/mL, 50ng/mL, 100ng/mL, 200ng/mL, with inner mark method ration, the interior scalar of addition is 5ng/mL, and by excellent
Condition measure after change establishes standard curve, and the regression equation and related coefficient for obtaining 7 kinds of sweeteners are shown in Table 3.
The linear dimensions of 3 standard curve of table
2.4 precision：
It is subject to 10ng/mL, 8 samples is often surveyed on the day of test sample and survey a 10ng/mL titers, while provide repetition in one day weekly
Ten 10ng/mL titers are surveyed, go out standard deviation with calculated by peak area：Saccharin sodium 3.02%, acesulfame-K 2.32%, honey element
2.68%, Aspartame 1.98%, neotame 2.89%, Sucralose 2.32%.
2.5 chromatogram
It is good with the condition measure 10ng/mL mixed standard solutions separating degree after optimization referring to Fig. 1, high sensitivity.
2.2.5 sample analysis
This time sample is taken to Wuhan City's lake water.
50mL water samples are taken, by 0.45 μm of membrane filtration, adjust PH=2 with dilute hydrochloric acid (analysis is pure) afterwards.With 5mL methanol
Oasis HLB solid phase extraction columns are activated with the ultra-pure water of 5mL (PH=2), then loading, after sample flow is complete, used
The ultra-pure water removal of impurities of 10mLPH=2, vacuum drain 30min, are finally eluted with the pure methanol of 4mL, by obtained eluent at 45 DEG C
Under conditions of nitrogen be blown to 0.5mL, use MeOH:H2O=25:75(V：V) constant volume finally centrifuges 10 points to 1mL with 12000rpm
Clock takes supernatant to carry out LC-MS/MS analyses.
The detection sample to be analyzed that takes that treated, is separated using C18 chromatographic columns, with H2O+5mM ammonium acetates+0.5mMTris
(trishydroxymethylaminomethane)-acetonitrile is mobile phase, gradient elution, by the component isolated of retention time and mass-to-charge ratio into
Row is qualitative, and the numerical value of 7 kinds of artificial sweetening agents is quantitatively obtained with internal standard method.
The one or more technical solutions provided in the embodiment of the present application, have at least the following technical effects or advantages：
The liquid chromatogram provided in the embodiment of the present application-triple quadrupole rods tandem mass spectrometries detect in environment water 7 kinds manually simultaneously
The method of sweetener using liquid chromatogram-triple quadrupole rods tandem mass spectrometries, to sample purify dense using solid phase extraction techniques
Contracting, can carry out trace analysis, and detection limits low, high sensitivity, and stability is good, contribute to more accurate, more accurately monitor water quality.
It should be noted last that more than specific embodiment is merely illustrative of the technical solution of the present invention and unrestricted, although
The present invention is described in detail with reference to example, it will be understood by those of ordinary skill in the art that, it can be to the skill of the present invention
Art scheme is modified or replaced equivalently, and without departing from the spirit and scope of technical solution of the present invention, should all be covered at this
Among the right of invention.
Claims (1)
1. the method that a kind of liquid chromatogram-triple quadrupole rods tandem mass spectrometries detect 7 kinds of artificial sweetening agents in environment water simultaneously,
It is characterized in that, including：
Standard items processing：
The standard items of powdered sweetener are taken respectively：Honey element CYC, saccharin SAC, acesulfame-K ACE, Aspartame ASP, trichlorine sugarcane
Sugared SUC, neotame NEO and neohesperidin dihydrochalcone NHDC are configured to the standard reserving solution of seven kinds of 1.00mg/mL with methanol；It will
Seven kinds of standard reserving solutions mix and seven kinds of standard mixed solutions of 1.00 μ g/mL are configured to methanol；
The internal standard standard items of powdered sweetener are taken respectively：ACE-d4, ASP-d5, SUC-d6, CYC-d11 are with 1:1 methanol/water
It is configured to 1.00 μ g/mL, tetra- kinds of internal standard mixed solutions；
It is 0.1ng/mL, 0.5ng/mL, 1ng/mL, 5ng/ that seven kinds of standard mixed solutions are configured to concentration gradient with methanol
The standard curve of mL, 10ng/mL, 50ng/mL, 100ng/mL, 200ng/mL；Wherein, seven kinds of marks of each concentration gradient
Equal tetra- kinds of internal standard mixed solutions containing 5ng/mL in quasi- mixed solution, for liquid chromatogram-triple quadrupole rods tandem mass spectrometries detection point
Analysis；
It is separated using C18 chromatographic columns, with H2O+5mMNH4Ac+0.5mMTris (trishydroxymethylaminomethane)-acetonitrile is stream
Dynamic phase, gradient elution is qualitative to each component progress with retention time and mass-to-charge ratio, and seven kinds of sweeteners are quantified with internal standard method, are obtained
To the regression equation and related coefficient of each sweetener；
Environmental water sample processing to be measured：
50mL environmental water samples are taken through 0.45 μm of membrane filtration, and the PH of the environmental water sample after filtering is adjusted to 2 with dilute hydrochloric acid；
Purifying concentration environmental water sample is carried out with Solid Phase Extraction：
5mL methanol is used respectively, and the 5mL ultra-pure waters of PH=2 activate Oasis HLB solid phase extraction columns；With per second 1 drop
Speed loading；It is eluted and cleaned with 10mL ultra-pure waters (PH=2), vacuum drains 30min；It is eluted with 4mL methanol, the elution that will be obtained
Liquid is blown to 0.5mL in 45 DEG C of nitrogen；It is 25 with volume ratio:75 methanol/water solution constant volume is to 1mL；12000rpm is centrifuged 10 minutes,
Supernatant is taken to carry out LC-MS/MS analyses；
It learns from else's experience the environment water body example of the environmental water sample processing step to be measured processing, is separated using C18 chromatographic columns, with H2O
+5mMNH4Ac+0.5mMTris (trishydroxymethylaminomethane)-acetonitrile be mobile phase, gradient elution, with retention time and matter lotus
It is more qualitative than being carried out for the component isolated, the concentration of seven kinds of sweeteners is obtained with internal standard method.
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| CN115254063A (en) * | 2022-08-26 | 2022-11-01 | 中国地质大学（武汉） | Gel adsorption film, gradient diffusion device and application in artificial sweetener sampling |
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