CN118067883A - Liquid chromatography detection method of dioxane - Google Patents
Liquid chromatography detection method of dioxane Download PDFInfo
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- CN118067883A CN118067883A CN202410401611.0A CN202410401611A CN118067883A CN 118067883 A CN118067883 A CN 118067883A CN 202410401611 A CN202410401611 A CN 202410401611A CN 118067883 A CN118067883 A CN 118067883A
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- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 title claims abstract description 145
- 238000001514 detection method Methods 0.000 title claims abstract description 112
- 238000004811 liquid chromatography Methods 0.000 title claims abstract description 41
- 239000000243 solution Substances 0.000 claims abstract description 63
- 238000000034 method Methods 0.000 claims abstract description 50
- 239000007788 liquid Substances 0.000 claims abstract description 45
- 239000012086 standard solution Substances 0.000 claims abstract description 33
- 239000007853 buffer solution Substances 0.000 claims abstract description 21
- 238000010829 isocratic elution Methods 0.000 claims abstract description 5
- 230000008569 process Effects 0.000 claims description 24
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 22
- 239000012498 ultrapure water Substances 0.000 claims description 22
- 230000002378 acidificating effect Effects 0.000 claims description 17
- 238000010828 elution Methods 0.000 claims description 13
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 12
- 238000002347 injection Methods 0.000 claims description 11
- 239000007924 injection Substances 0.000 claims description 11
- 239000000872 buffer Substances 0.000 claims description 9
- QAOWNCQODCNURD-UHFFFAOYSA-M hydrogensulfate Chemical compound OS([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-M 0.000 claims description 8
- 239000010413 mother solution Substances 0.000 claims description 8
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 claims description 7
- YBCVMFKXIKNREZ-UHFFFAOYSA-N acoh acetic acid Chemical compound CC(O)=O.CC(O)=O YBCVMFKXIKNREZ-UHFFFAOYSA-N 0.000 claims description 6
- XVVLAOSRANDVDB-UHFFFAOYSA-N formic acid Chemical compound OC=O.OC=O XVVLAOSRANDVDB-UHFFFAOYSA-N 0.000 claims description 6
- 239000000945 filler Substances 0.000 claims description 4
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 4
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 4
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 4
- 239000000377 silicon dioxide Substances 0.000 claims description 4
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 3
- WBHQBSYUUJJSRZ-UHFFFAOYSA-M sodium bisulfate Chemical compound [Na+].OS([O-])(=O)=O WBHQBSYUUJJSRZ-UHFFFAOYSA-M 0.000 claims description 3
- 229910000342 sodium bisulfate Inorganic materials 0.000 claims description 3
- JTNCEQNHURODLX-UHFFFAOYSA-N 2-phenylethanimidamide Chemical compound NC(=N)CC1=CC=CC=C1 JTNCEQNHURODLX-UHFFFAOYSA-N 0.000 claims description 2
- ADCGAPKUMAQOLJ-UHFFFAOYSA-N azane;formic acid Chemical compound N.OC=O.OC=O ADCGAPKUMAQOLJ-UHFFFAOYSA-N 0.000 claims description 2
- ALSPKRWQCLSJLV-UHFFFAOYSA-N azanium;acetic acid;acetate Chemical compound [NH4+].CC(O)=O.CC([O-])=O ALSPKRWQCLSJLV-UHFFFAOYSA-N 0.000 claims description 2
- YYRMJZQKEFZXMX-UHFFFAOYSA-L calcium bis(dihydrogenphosphate) Chemical compound [Ca+2].OP(O)([O-])=O.OP(O)([O-])=O YYRMJZQKEFZXMX-UHFFFAOYSA-L 0.000 claims description 2
- 229910000389 calcium phosphate Inorganic materials 0.000 claims description 2
- JXRVKYBCWUJJBP-UHFFFAOYSA-L calcium;hydrogen sulfate Chemical compound [Ca+2].OS([O-])(=O)=O.OS([O-])(=O)=O JXRVKYBCWUJJBP-UHFFFAOYSA-L 0.000 claims description 2
- 238000007872 degassing Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 235000019691 monocalcium phosphate Nutrition 0.000 claims description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 2
- 229910000343 potassium bisulfate Inorganic materials 0.000 claims description 2
- 238000009210 therapy by ultrasound Methods 0.000 claims description 2
- 229940062672 calcium dihydrogen phosphate Drugs 0.000 claims 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims 1
- 230000014759 maintenance of location Effects 0.000 abstract description 18
- 230000008859 change Effects 0.000 abstract description 7
- 230000002035 prolonged effect Effects 0.000 abstract description 5
- 239000000523 sample Substances 0.000 description 30
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 27
- 230000000052 comparative effect Effects 0.000 description 12
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000012360 testing method Methods 0.000 description 5
- 239000012496 blank sample Substances 0.000 description 4
- 238000004364 calculation method Methods 0.000 description 4
- 238000011835 investigation Methods 0.000 description 4
- 239000012452 mother liquor Substances 0.000 description 4
- 239000012488 sample solution Substances 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 239000001569 carbon dioxide Substances 0.000 description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- JHUUPUMBZGWODW-UHFFFAOYSA-N 3,6-dihydro-1,2-dioxine Chemical compound C1OOCC=C1 JHUUPUMBZGWODW-UHFFFAOYSA-N 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-M Formate Chemical compound [O-]C=O BDAGIHXWWSANSR-UHFFFAOYSA-M 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical compound [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- -1 medical products Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 210000002345 respiratory system Anatomy 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 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 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 231100000570 acute poisoning Toxicity 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229940127554 medical product Drugs 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229940111688 monobasic potassium phosphate Drugs 0.000 description 1
- 229940045641 monobasic sodium phosphate Drugs 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000000611 regression analysis Methods 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 239000002966 varnish Substances 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The application belongs to the technical field of liquid chromatography detection, and particularly relates to a liquid chromatography detection method of dioxane; the liquid chromatography detection method of dioxane provided by the application comprises the steps of sequentially injecting a group of dioxane standard solution and sample liquid to be detected containing dioxane into a high performance liquid chromatograph for detection; and the detection method is provided with the elution program of a liquid chromatograph as follows: the method takes the acid buffer solution as the mobile phase to perform isocratic elution, the pH value of the weak acid buffer solution is difficult to change, the retention time of the dioxane detection is consistent, the repeatability of the detection result is good, meanwhile, the retention time of the 1, 4-dioxane can be prolonged, the tailing of chromatographic peaks is obviously improved, the detection result has better repeatability, specificity, precision and accuracy, and therefore the technical problem that the detection is not ideal due to the mobile phase used in the liquid chromatography detection method of the dioxane in the prior art is solved.
Description
Technical Field
The application belongs to the technical field of liquid chromatography detection, and particularly relates to a liquid chromatography detection method of dioxane.
Background
1,4 Dioxane is an organic compound, also called 1, 4-dioxane, with a molecular formula of C 4H8O2, and is colorless liquid with slight fragrance; 1,4 dioxane is a widely used chemical, the most common application is solvents, emulsifiers, detergents and the like, the 1,4 dioxane can also be used for producing pesticides, medical products, dyes, cellulose acetate, resins, vegetable oils, mineral oil and the like, and can also be used for the production process of paint, varnish, plasticizer, wetting agent, perfume and the like, and the wide use of 1,4 dioxane leads to the easy leakage of the 1,4 dioxane into the natural environment; when a human body contacts the 1,4 dioxane through skin, respiratory tract, digestive tract and other ways, the human body has irritation to the skin, eyes and respiratory system, and after entering the human body, damage to liver, kidney and nervous system can be caused, even death caused by acute poisoning and other conditions occur, and the 1,4 dioxane is a dangerous chemical which can be cancerogenic, so that the 1,4 dioxane needs to be detected to control the hazard.
At present, the detection of the 1,4 dioxane is mainly a liquid chromatography detection method, mainly a gas chromatography detection method, and although the detection limit and the accuracy can be lower, the use of an instrument is expensive, the operation flow is complex, and the water sample product containing interference components formed in the processing process of the medical instrument is difficult to accurately detect; in the detection process, ultrapure water and/or acetonitrile is often used as a mobile phase, and for the mobile phase of ultrapure water, since the ultrapure water almost contains no ions, is usually neutral (pH=7), carbon dioxide in the air is easily absorbed by the air and is changed into weak acidity by exposure to the air, the pH value of the mobile phase of ultrapure water changes, and the 1,4 dioxane is a pH sensitive compound, and the tiny pH change in the liquid chromatography test process can lead to unstable environment-friendly retention time of the dioxane and poor repeatability; when pure water and acetonitrile are used as mobile phases, the retention time of the 1,4 dioxane is short, the tailing phenomenon of chromatographic peaks is easy to occur, the precision and the accuracy of detection results are low, and the detection requirements of medical instrument processing still cannot be met. Therefore, there is a need for an improvement in the current liquid chromatography detection methods for dioxane.
Disclosure of Invention
In view of the above, the present application provides a liquid chromatography detection method for dioxane, which is used for solving the technical problem of non-ideal detection caused by mobile phase used in the liquid chromatography detection method for dioxane in the prior art.
The first aspect of the application provides a liquid chromatography detection method of dioxane, comprising the following steps:
Injecting the sample liquid to be detected containing the dioxane into a liquid chromatograph with chromatographic parameters, and carrying out liquid chromatograph detection to obtain a detection result of the sample liquid to be detected;
among the chromatographic parameters:
The elution procedure was set to perform an isocratic elution with acidic buffer as mobile phase.
Preferably, the pH of the acidic buffer is from 2 to 6.
Preferably, the acidic buffer is selected from at least one of a dihydrogen phosphate solution, a hydrogen sulfate solution, an acetic acid-acetate solution, and a formic acid-formate solution.
Preferably, the monobasic phosphate solution is selected from a monobasic sodium phosphate solution, a monobasic potassium phosphate solution or a monobasic calcium phosphate solution;
The bisulfate solution is selected from sodium bisulfate solution, potassium bisulfate solution or calcium bisulfate solution;
The acetic acid-acetate solution is selected from acetic acid-ammonium acetate solution;
the formic acid-formate solution is selected from formic acid-ammonium formate solutions.
Preferably, the concentration of the dihydrogen phosphate solution is 0.005-0.1 mol/L;
The concentration of the bisulfate solution is 0.005-0.1 mol/L;
The concentration of the acetic acid-acetate solution is 0.005-0.1 mol/L;
the concentration of the formic acid-formate solution is 0.005-0.1 mol/L.
Preferably, the acidic buffer solution is subjected to pretreatment, and the pretreatment step includes: filtering the acid buffer solution by a filter membrane with the aperture of 0.22 mu m, and degassing by ultrasonic treatment for 15-20 min.
Preferably, among the chromatographic parameters:
The chromatographic column is a chromatographic column taking octadecylsilane chemically bonded silica as a filler;
The detection wavelength is 190-210 nm;
The flow rate is 0.5-1.0 mL/min;
the column temperature is 25-40 ℃;
the sample injection amount is 10-20 mu L.
Preferably, before injecting the sample solution to be tested containing dioxane into the high performance liquid chromatograph with the chromatographic parameters set, the method further comprises:
injecting a group of dioxane standard solutions with known concentrations into a high performance liquid chromatograph with chromatographic parameters set, and performing liquid chromatography detection to obtain detection results of the group of dioxane standard solutions;
And establishing a standard curve of the dioxane according to the concentration and the detection result of a group of dioxane standard solutions.
Among the chromatographic parameters:
The chromatographic column is a chromatographic column taking octadecylsilane chemically bonded silica as a filler;
the mobile phase is an acidic buffer solution;
The detection wavelength is 190-210 nm;
The flow rate is 0.5-1.0 mL/min;
the column temperature is 25-40 ℃;
the sample injection amount is 10-20 mu L.
Preferably, the set of dioxane standard solutions comprises dioxane standard solutions with concentration of 2.0 mug/mL, 3.0 mug/mL, 4.0 mug/mL, 6.0 mug/mL, 12.0 mug/mL and 24.0 mug/mL.
Preferably, the configuration process of the set of dioxane standard solutions comprises:
A set of dioxane standard solutions with concentrations of 2.0. Mu.g/mL, 3.0. Mu.g/mL, 4.0. Mu.g/mL, 6.0. Mu.g/mL, 12.0. Mu.g/mL, 24.0. Mu.g/mL were obtained by pipetting 1.0mL, 1.5mL, 2.0mL, 3.0mL, 6.0mL, 12.0mL into a 50mL volumetric flask with a pipette or pipette, diluting with ultrapure water, and metering to the scale.
In summary, the application provides a liquid chromatography detection method of dioxane, which comprises setting chromatographic parameters of a high performance liquid chromatograph, wherein mobile phase in the chromatographic parameters is an acidic buffer solution; compared with the method which uses pure water as a mobile phase and easily absorbs carbon dioxide to change the pH value from neutral to weak acid during liquid chromatography detection, the method has the advantages that the acid buffer solution is used as the mobile phase, and has certain acidity, so that the pH value is difficult to change during detection, the property of the mobile phase is stabilized, the stability of the environment-friendly retention time of dioxane is improved, the dioxane can peak at a fixed position, the repeatability of a detection result is good, and the qualitative detection of dioxane is facilitated; meanwhile, compared with pure water and acetonitrile serving as mobile phases, the method has the advantages that the peak is easy to occur early in a dioxane environment-friendly retention period during liquid chromatography detection, and the retention time of a1, 4-dioxane is prolonged by using the acidic buffer solution serving as the mobile phases, so that the tailing of the chromatographic peaks is remarkably improved, the peak area calculation accuracy of the chromatographic peaks is high, the accurate quantitative detection of the 1, 4-dioxane is facilitated, the detection result has good specificity, precision and accuracy, and the technical problem of non-ideal detection caused by the mobile phases used in the liquid chromatography detection method of the dioxane in the prior art is solved.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the following description will briefly explain the drawings needed in the embodiments or the prior art, and it is obvious that the drawings in the following description are some embodiments of the present application and that other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a standard working curve of a set of dioxane standard solutions in example 1 of the present application;
FIG. 2 is a chromatogram obtained by the test of a blank sample of ultrapure water in example 3 of the present application;
FIG. 3 is a chromatogram obtained from a standard stock 1, 4-dioxane mother liquor test in example 3 of the present application;
FIG. 4 is a chromatogram obtained by detecting 1, 4-dioxane by the liquid chromatography detection method provided in example 1 in example 4 of the present application;
FIG. 5 is a chromatogram obtained by detecting 1, 4-dioxane by a second sample injection of the liquid chromatography detection method provided in example 1 in example 4 of the present application;
FIG. 6 is a chromatogram obtained by detecting 1, 4-dioxane by first sample injection using the liquid chromatography detection method provided in comparative example 1 in example 4 of the present application;
FIG. 7 is a chromatogram obtained by detecting 1, 4-dioxane by a second sample injection using the liquid chromatography detection method provided in comparative example 1 in example 4 of the present application;
FIG. 8 is a chromatogram obtained by detecting 1, 4-dioxane according to the liquid chromatography detection method provided in comparative example 2 in example 4 of the present application.
Detailed Description
The application provides a liquid chromatography detection method of dioxane, which is used for solving the technical problem of non-ideal detection caused by mobile phase used in the liquid chromatography detection method of dioxane in the prior art.
The following description of the embodiments of the present application will be made apparent and fully in view of the accompanying drawings, in which some, but not all embodiments of the application are shown. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
In view of the defect that the liquid chromatogram of the existing liquid chromatogram by adopting ultrapure water (acetonitrile) as a mobile phase is not ideal, the application provides a liquid chromatogram detection method of dioxane, which comprises the steps of injecting a sample liquid to be detected containing dioxane into a high performance liquid chromatograph with chromatographic parameters, and carrying out liquid chromatogram detection to obtain a detection result of the sample liquid to be detected; wherein, among the chromatographic parameters: the elution procedure is set to take acid buffer as mobile phase to carry out isocratic elution; when ultrapure water is used as a mobile phase for elution in the prior art, as the ultrapure water almost contains ions and is neutral, carbon dioxide in the air can be absorbed to cause the change of pH value, the pH value is reduced to be slightly acidic, dioxane is a pH sensitive compound, the change of the pH value can cause inconsistent retention time of the dioxane detected before and after, the detection result is difficult to reproduce, the qualitative detection of the dioxane is not facilitated, and the application adopts a weak acid buffer solution with certain acidity as the mobile phase, the pH value of the weak acid buffer solution is difficult to change, so that the retention time of the dioxane detected before and after is consistent, the repeatability of the detection result is good, and the qualitative detection of the dioxane is facilitated; meanwhile, compared with the ultrapure water and acetonitrile which are used as mobile phases for elution, the acidic buffer solution used by the application is used as the mobile phase, so that the retention time of the 1, 4-dioxane can be increased, the tailing of chromatographic peaks is obviously improved, and the defect of non-ideal detection caused by the ultrapure water (acetonitrile) mobile phase used by the existing liquid chromatography detection method of the dioxane is overcome.
Preferably, the pH of the acidic buffer used in the present application is in the range of 2 to 6, and since the pH is too low, the column is liable to be damaged, and the acidic buffer having too low pH is not used in the present application.
Preferably, the acidic buffer selected in the present application is selected from buffers such as dihydrogen phosphate solution, bisulfate solution, formate solution, etc., dihydrogen phosphate such as sodium dihydrogen phosphate, bisulfate such as sodium bisulfate, etc.; meanwhile, when acetate solution such as ammonium acetate or formate solution such as ammonium formate is used, acetic acid or formic acid is added to adjust the pH to be weak acid, and the pH range is controlled to be 2-6.
Preferably, the method also pretreats the acid buffer solution, and the pretreatment process is to filter the acid buffer solution through a filter membrane with the aperture of 0.22 mu m, and degasify the acid buffer solution for 15-20 min.
As a further preferred aspect, the present application further includes injecting a set of dioxane standard solutions with known concentrations into the high performance liquid chromatograph with set chromatographic parameters before injecting the sample solution to be detected containing dioxane into the liquid chromatograph for detection, and establishing a standard curve according to the detection results and the corresponding concentrations of the set of dioxane standard solutions, and then injecting the sample solution to be detected containing dioxane into the liquid chromatograph for liquid chromatography, wherein the accurate concentration of dioxane in the sample solution to be detected can be obtained due to the pre-established standard curve, thereby realizing quantitative detection;
In the detection process, chromatographic parameters set by a group of dioxane standard solutions and sample liquid to be detected containing dioxane are the same, an acid buffer solution is used as a mobile phase for isocratic elution in the chromatographic parameters, and compared with pure water and acetonitrile which are used as mobile phases for elution, the acid buffer solution is used for elution in the application, so that the retention time of 1, 4-dioxane can be prolonged, the tailing of chromatographic peaks is obviously improved, meanwhile, interference peaks can not appear near the chromatographic peaks of 1, 4-dioxane, the accurate calculation of the chromatographic peak area or peak height of the 1, 4-dioxane with high, medium and low concentration is facilitated, and the quantitative detection result of the 1, 4-dioxane is excellent in accuracy and precision.
Preferably, the preparation process of the dioxane standard solution comprises the steps of sucking standard reserve mother solutions with different volumes into a group of volumetric flasks, diluting with ultrapure water, and fixing the volume to a scale to obtain a group of dioxane standard solution; in order to improve the accuracy of the detection result, the standard stock mother solution can be prepared for use at present, the commercial dioxane standard solution can be purchased directly, and the standard stock mother solution can be absorbed by a pipetting gun with good accuracy or by a traditional ear-washing ball matched with a pipette.
Example 1
The embodiment 1 of the application provides a liquid chromatography detection method of dioxane, which comprises the following steps: preparing standard stock mother solution, preparing a group of dioxane standard solution, preparing standard adding detection solution, establishing a standard curve and detecting dioxane.
Wherein, the process of preparing standard stock mother liquor comprises: precisely weighing 0.500g of 1, 4-dioxane in a 500mL volumetric flask, adding ultrapure water for dissolution, fixing the volume to a scale, and shaking uniformly to obtain a standard mother solution with the concentration of 1000 mug/mL. Taking 5.00mL of 1, 4-dioxane standard mother liquor, diluting with ultrapure water, fixing the volume to a scale, and shaking uniformly to obtain standard stock mother liquor with the concentration of 100 mug/mL.
The process of preparing a set of dioxane standard solutions includes: 1.0, 1.5, 2.0, 3.0, 6.0 and 12.0mL of the standard stock solution are accurately sucked respectively, placed in a 50mL volumetric flask, diluted to scale by ultrapure water, and a group of dioxane standard solutions with the concentration of 1, 4-dioxane of 2.0, 3.0, 4.0, 6.0, 12.0 and 24.0 mug/mL are prepared.
The process for preparing the labeling detection liquid comprises the following steps: standard stock solution of dioxane with the concentration of 100 mug/mL is used as the adding standard solution. A certain amount of the standard solution is taken according to the table 1 and added into a 5mL centrifuge tube, the sample leaching solution is added to prepare the standard detection solutions with three concentrations of C1, C2 and C3, and 3 parallel samples are prepared for each concentration, and the standard detection solutions are shown in the table 1.
| Theoretical concentration | Adding the concentration of the standard solution | Adding the volume of the standard solution | Volume of leach liquor | Total volume of |
| C 1 (about 2.0 μg/mL) | 100μg/mL | 100μL | 4900μL | 5mL |
| C 2 (about 6.0 μg/mL) | 100μg/mL | 300μL | 4700μL | 5mL |
| C 3 (about 10.0 μg/mL) | 100μg/mL | 500μL | 4500μL | 5mL |
TABLE 1
The process of establishing the standard curve comprises the following steps: precisely measuring a group of dioxane standard solutions, injecting the dioxane standard solutions into a high performance liquid chromatograph for analysis, recording a chromatogram, and carrying out regression analysis on the obtained peak areas and the solubility of the corresponding standard working solutions to obtain a 1, 4-dioxane standard curve; the standard working curves for a set of dioxane standard solutions are shown in figure 1.
The process of detecting dioxane comprises the following steps: precisely measuring a sample liquid to be measured, injecting the sample liquid into a high performance liquid chromatograph, analyzing, recording a chromatogram, analyzing the attribution of each peak in the chromatogram, and judging whether the sample liquid to be measured contains a1, 4-dioxane or not;
Quantitative analysis: according to the peak area or peak height of the 1, 4-dioxane in the spectrogram of the sample to be detected, the content of the 1, 4-dioxane in the sample liquid to be detected is calculated by an external standard method by contrasting the standard working curve of a group of dioxane standard solutions.
The instrument used in the process of establishing the standard curve and the process of detecting the dioxane is a high performance liquid chromatograph with an ultraviolet detector, and the chromatographic parameters are as follows:
Chromatographic column: ZORBAX Eclipse plus C184.6.6 mm. Times.250 mm,5 μm;
mobile phase: 0.01mol/L sodium dihydrogen phosphate aqueous solution;
Detection wavelength: 200nm;
flow rate: 1.0mL/min;
Column temperature: 40 ℃;
Sample injection amount: 20. Mu.L;
Elution time: 15min.
Example 2
Embodiment 2 of the present application provides a liquid chromatography detection method for dioxane, which is different from embodiment 1 in that chromatographic parameters of a process of establishing a standard curve and a process of detecting dioxane are adjusted, and the chromatographic parameters set in embodiment 2 are as follows:
Chromatographic column: ZORBAX Eclipse plus C184.6.6 mm. Times.250 mm,5 μm;
Mobile phase: 0.01mol/L ammonium formate;
Detection wavelength: 190nm;
Flow rate: 0.5mL/min;
column temperature: 25 ℃;
sample injection amount: 10. Mu.L;
Elution time: 15min.
Comparative example 1
The comparative example 1 of the present application provides a liquid chromatography detection method of dioxane, which is different from example 1 in that the chromatographic parameters of the process of establishing a standard curve and the process of detecting dioxane are adjusted, and the chromatographic parameters are:
Chromatographic column: ZORBAX Eclipse plus C184.6.6 mm. Times.250 mm,5 μm;
Mobile phase: ultrapure water;
Detection wavelength: 200nm;
flow rate: 1.0mL/min;
Column temperature: 40 ℃;
Sample injection amount: 20. Mu.L;
Elution time: and 10min.
Comparative example 2
The comparative example 2 of the present application provides a liquid chromatography detection method of dioxane, which is different from example 1 in that the chromatographic parameters of the process of establishing a standard curve and the process of detecting dioxane are adjusted, and the chromatographic parameters are:
Chromatographic column: ZORBAX Eclipse plus C184.6.6 mm. Times.250 mm,5 μm;
Mobile phase: mobile phase a was ultrapure water and mobile phase B was acetonitrile, and gradient elution was performed according to table 2;
Detection wavelength: 200nm;
flow rate: 1.0mL/min;
Column temperature: 40 ℃;
Sample injection amount: 20. Mu.L.
| Time/min | Mobile phase A/v% | Mobile phase B/v% |
| 0 | 100 | 0 |
| 5 | 100 | 0 |
| 6 | 0 | 100 |
| 7 | 0 | 100 |
| 8 | 100 | 0 |
| 13 | 100 | 0 |
TABLE 2
Example 3
The method for detecting the dioxane liquid chromatography provided by the application is examined in a methodology, and comprises specificity, detection limit, quantitative limit, precision and accuracy.
The investigation process of the specificity comprises the following steps: the liquid chromatography detection method of dioxane provided by the application is adopted to measure the blank sample of ultrapure water and the standard stock mother solution of dioxane with the concentration of 100 mug/mL, and the chromatograms of the blank sample of ultrapure water and the standard stock mother solution with the concentration of 100 mug/mL are compared; from fig. 2 to 3, it can be seen that the dioxane showed a peak around 9.7min, and the blank sample showed no interference peak here, indicating that the method has good specificity for dioxane.
The inspection process of the detection limit and the quantitative limit comprises the following steps: since the solvent ultrapure water does not interfere with the measurement of the dioxane content, the detection limit and the quantitative limit are measured by a signal-to-noise ratio (S/N) method, and the standard stock solution is diluted by the ultrapure water, and the signal-to-noise ratio (S/N) is 3: the sample concentration at about 1 is the detection limit, and the signal to noise ratio is (S/N) 10: the sample concentration at about 1 is the quantitative limit. The detection limit of the method is 1.5 mug/mL (S/N is 6.62), and the quantitative limit is 2 mug/mL (S/N is 11.41).
The precision investigation process comprises the following steps: the liquid chromatographic detection method of dioxane provided by the application is used for measuring 9 low, medium and high concentration standard adding detection liquids (3 parallel samples, each parallel sample comprises the standard adding detection liquids with low, medium and high concentrations) provided in the embodiment 1, and recording chromatograms, and the calculated results are summarized in the table 3, and the average value of the Relative Standard Deviation (RSD) of the low, medium and high concentrations of dioxane is 0.89 percent and less than 6 percent, which indicates that the precision is good.
TABLE 3 Table 3
The investigation process of the accuracy comprises the following steps: the liquid chromatographic detection method for dioxane provided by the application is used for measuring 9 low, medium and high concentration standard adding detection liquids (3 parallel samples, wherein each parallel sample comprises the standard adding detection liquids with low, medium and high concentrations) in the embodiment 1, recording chromatograms, summarizing calculation results in table 4, comparing the measured concentration with the theoretical concentration from table 4, and calculating to obtain that the recovery rate of 1, 4-dioxane in the leaching liquid of a sample to be detected is in the range of 97% -108%, thus indicating that the accuracy meets the requirements.
Example 4
This example 4 is a view of the method advancement of the liquid chromatography detection method for dioxane provided in example 1 and comparative examples 1 to 2.
The method advanced investigation process comprises the following steps: the standard stock solutions of 1, 4-dioxane were tested with the liquid chromatography detection methods for dioxane provided in example 1, comparative example 2, respectively; meanwhile, referring also to example 3, the precision and accuracy test results of the liquid chromatography test method for dioxane provided in comparative examples 1-2 were obtained, wherein the test results are shown in fig. 4-7 and table 4.
TABLE 4 Table 4
As can be seen from comparing fig. 4 to 5, fig. 6 to 7, and table 4, in the chromatograms obtained by the first and second detection methods for dioxane provided in comparative example 1, the retention time of dioxane is short, the retention time of dioxane is inconsistent, the detection result is difficult to reproduce, and in the chromatograms obtained by the first and second detection methods for dioxane provided in example 1, the retention time of dioxane is prolonged, the retention time of dioxane is consistent, the detection result can be reproduced, and qualitative detection of dioxane is facilitated.
As can be seen from table 4 and fig. 8, in the detection result of the liquid chromatography detection method using dioxane provided in comparative example 2, the retention time of dioxane is shorter, 5.98min, the tailing factor is large, 1.713, the average value of RSD% of the detection sample is close to 6%, 4.40%, the recovery rate range of the detection sample is larger, 82% -113%, while as can be seen from table 4 and fig. 4, the retention time of dioxane is longer, 9.729, the tailing factor is small, 1.19, the average value of RSD% of the detection sample is smaller than 1%, 0.89%, the recovery rate range of the detection sample is smaller, 97% -108%, which indicates that compared with the use of pure water and acetonitrile as mobile phases, the retention time of 1, 4-dioxane can be prolonged, the tailing area calculation accuracy of the chromatographic peak is remarkably improved, and the accuracy of the specific determination of 1, 4-dioxane detection result is good, and accuracy.
The above embodiments are only for illustrating the technical solution of the present application, and not for limiting the same; although the application has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the application.
Claims (10)
1. A method for detecting dioxane by liquid chromatography, comprising:
Injecting the sample liquid to be detected containing the dioxane into a liquid chromatograph with chromatographic parameters, and carrying out liquid chromatograph detection to obtain a detection result of the sample liquid to be detected;
among the chromatographic parameters:
The elution procedure is an isocratic elution with an acidic buffer as mobile phase.
2. The method for detecting dioxane according to claim 1, wherein the pH of the acidic buffer solution is=2 to 6.
3. The method for detecting dioxane according to claim 1, wherein the acidic buffer is at least one selected from the group consisting of a dihydrogen phosphate solution, a hydrogen sulfate solution, an acetic acid-acetate solution, and a formic acid-formate solution.
4. A method for liquid chromatographic detection of dioxane according to claim 3 wherein the dihydrogen phosphate solution is selected from the group consisting of sodium dihydrogen phosphate solution, potassium dihydrogen phosphate solution, and calcium dihydrogen phosphate solution;
The bisulfate solution is selected from sodium bisulfate solution, potassium bisulfate solution or calcium bisulfate solution;
The acetic acid-acetate solution is selected from acetic acid-ammonium acetate solution;
the formic acid-formate solution is selected from formic acid-ammonium formate solutions.
5. The method for detecting dioxane according to claim 3, wherein the concentration of the dihydrogen phosphate solution is 0.005-0.1 mol/L;
The concentration of the bisulfate solution is 0.005-0.1 mol/L;
The concentration of the acetic acid-acetate solution is 0.005-0.1 mol/L;
the concentration of the formic acid-formate solution is 0.005-0.1 mol/L.
6. The method for detecting dioxane according to claim 1, wherein the acidic buffer solution is subjected to pretreatment, and the pretreatment step comprises: filtering the acid buffer solution by a filter membrane with the aperture of 0.22 mu m, and degassing by ultrasonic treatment for 15-20 min.
7. The method for detecting dioxane according to claim 1, wherein the chromatographic parameters are as follows:
The chromatographic column is a chromatographic column taking octadecylsilane chemically bonded silica as a filler;
The detection wavelength is 190-210 nm;
The flow rate is 0.5-1.0 mL/min;
the column temperature is 25-40 ℃;
The sample injection amount is 10-20 mu L;
The elution time is 10-15 min.
8. The method for detecting liquid chromatography using dioxane according to claim 1, wherein before injecting the sample liquid to be detected containing dioxane into the high performance liquid chromatograph with chromatographic parameters set, further comprising:
injecting a group of dioxane standard solutions with known concentrations into a high performance liquid chromatograph with chromatographic parameters set, and performing liquid chromatography detection to obtain detection results of the group of dioxane standard solutions;
establishing a standard curve of the dioxane according to the concentration and the detection result of a group of dioxane standard solutions;
Among the chromatographic parameters:
The chromatographic column is a chromatographic column taking octadecylsilane chemically bonded silica as a filler;
the mobile phase is an acidic buffer solution;
The detection wavelength is 190-210 nm;
The flow rate is 0.5-1.0 mL/min;
the column temperature is 25-40 ℃;
The sample injection amount is 10-20 mu L;
The elution time is 10-15 min.
9. The method according to claim 8, wherein the set of dioxane standard solutions comprises dioxane standard solutions having a concentration of 2.0 μg/mL, 3.0 μg/mL, 4.0 μg/mL, 6.0 μg/mL, 12.0 μg/mL, and 24.0 μg/mL.
10. The method for detecting dioxane according to claim 9, wherein the configuration process of the set of dioxane standard solutions comprises:
1.0mL, 1.5mL, 2.0mL, 3.0mL, 6.0mL and 12.0mL of standard stock mother solution with the concentration of 100 mug/mL are sucked into a 50mL volumetric flask, diluted with ultrapure water and fixed to volume to a scale, and a group of dioxane standard solutions with the concentration of 2.0 mug/mL, 3.0 mug/mL, 4.0 mug/mL, 6.0 mug/mL, 12.0 mug/mL and 24.0 mug/mL are obtained.
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