CN115326942A - Analytical method for measuring p-toluenesulfonic acid thiophene ester - Google Patents
Analytical method for measuring p-toluenesulfonic acid thiophene ester Download PDFInfo
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- CN115326942A CN115326942A CN202210161316.3A CN202210161316A CN115326942A CN 115326942 A CN115326942 A CN 115326942A CN 202210161316 A CN202210161316 A CN 202210161316A CN 115326942 A CN115326942 A CN 115326942A
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- mobile phase
- toluenesulfonate
- analytical method
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- thiophene
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- 238000004458 analytical method Methods 0.000 title claims abstract description 17
- -1 p-toluenesulfonic acid thiophene ester Chemical class 0.000 title claims description 19
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 claims abstract description 17
- 238000010828 elution Methods 0.000 claims abstract description 16
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 125000005233 alkylalcohol group Chemical group 0.000 claims abstract description 6
- 239000007788 liquid Substances 0.000 claims abstract description 5
- 239000000945 filler Substances 0.000 claims abstract description 4
- 239000002253 acid Substances 0.000 claims abstract description 3
- 239000007864 aqueous solution Substances 0.000 claims abstract description 3
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 3
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 3
- 150000002500 ions Chemical class 0.000 claims description 29
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- 238000001514 detection method Methods 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 13
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 8
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 238000011002 quantification Methods 0.000 claims 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 claims description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 4
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 4
- 235000019253 formic acid Nutrition 0.000 claims description 4
- ITBQALJLFZKJKR-UHFFFAOYSA-N 4-methylbenzenesulfonic acid;thiophene Chemical compound C=1C=CSC=1.CC1=CC=C(S(O)(=O)=O)C=C1 ITBQALJLFZKJKR-UHFFFAOYSA-N 0.000 claims description 3
- 239000011260 aqueous acid Substances 0.000 claims description 3
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 claims description 3
- 238000012512 characterization method Methods 0.000 claims description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 2
- 238000003556 assay Methods 0.000 claims 2
- 238000011895 specific detection Methods 0.000 abstract description 2
- 239000007789 gas Substances 0.000 description 12
- 239000012535 impurity Substances 0.000 description 7
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 6
- 229950010477 clopidogrel hydrogen sulphate Drugs 0.000 description 6
- FDEODCTUSIWGLK-UHFFFAOYSA-N hydrogen sulfate;hydron;methyl 2-(2-chlorophenyl)-2-(6,7-dihydro-4h-thieno[3,2-c]pyridin-5-yl)acetate Chemical compound OS(O)(=O)=O.C1CC=2SC=CC=2CN1C(C(=O)OC)C1=CC=CC=C1Cl FDEODCTUSIWGLK-UHFFFAOYSA-N 0.000 description 6
- 238000001035 drying Methods 0.000 description 4
- 238000007865 diluting Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 230000000702 anti-platelet effect Effects 0.000 description 2
- 239000003146 anticoagulant agent Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 231100000025 genetic toxicology Toxicity 0.000 description 2
- 230000001738 genotoxic effect Effects 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 2
- 125000004182 2-chlorophenyl group Chemical group [H]C1=C([H])C(Cl)=C(*)C([H])=C1[H] 0.000 description 1
- ZDMWSYXPZCVAOH-UHFFFAOYSA-N C(=O)(O)C(O)C(O)C(=O)O.COC(CN)=O Chemical compound C(=O)(O)C(O)C(O)C(=O)O.COC(CN)=O ZDMWSYXPZCVAOH-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- MVPPADPHJFYWMZ-UHFFFAOYSA-N chlorobenzene Chemical compound ClC1=CC=CC=C1 MVPPADPHJFYWMZ-UHFFFAOYSA-N 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- FDEODCTUSIWGLK-RSAXXLAASA-N clopidogrel sulfate Chemical compound [H+].OS([O-])(=O)=O.C1([C@H](N2CC=3C=CSC=3CC2)C(=O)OC)=CC=CC=C1Cl FDEODCTUSIWGLK-RSAXXLAASA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- FUKUFMFMCZIRNT-UHFFFAOYSA-N hydron;methanol;chloride Chemical compound Cl.OC FUKUFMFMCZIRNT-UHFFFAOYSA-N 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- QYSXFLDJHDDOLS-UHFFFAOYSA-N methyl acetate;sulfuric acid Chemical compound COC(C)=O.OS(O)(=O)=O QYSXFLDJHDDOLS-UHFFFAOYSA-N 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- JRMUNVKIHCOMHV-UHFFFAOYSA-M tetrabutylammonium bromide Chemical compound [Br-].CCCC[N+](CCCC)(CCCC)CCCC JRMUNVKIHCOMHV-UHFFFAOYSA-M 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- 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
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- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
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Abstract
The invention provides an analysis method for measuring p-toluenesulfonate, which adopts a liquid chromatography-mass spectrometer to detect and analyze p-toluenesulfonate, adopts a chromatographic column with octadecylsilane chemically bonded silica as a filler, takes an acid aqueous solution as a mobile phase A, takes lower alkyl alcohol or acetonitrile as a mobile phase B, and has the volume concentration of 0.1-0.5 percent and the volume fraction of 25-50 percent; gradient elution was performed. The invention provides an accurate, efficient and highly specific detection and analysis method for the ppm level determination of p-toluenesulfonate.
Description
Technical Field
The invention relates to the technical field of drug detection and analysis, in particular to an analysis method for determining p-toluenesulfonate.
Background
The chemical name of clopidogrel hydrogen sulfate is: s (+) -2- (2-chlorophenyl) -2- (4,5,6,7-tetrahydrothiophene [3,2-c]And pyridine-5) methyl acetate hydrogensulfate salt ofThe anti-platelet aggregation drug is developed by Sanofi company of France in 1986, and is a novel efficient anti-platelet aggregation drug. The CAS number is: 120202-66-6, molecular formula C 16 H 16 ClNO 2 ·H 2 SO 4 The molecular weight is: 419.90, the structural formula is as follows:
the main synthesis reaction process of clopidogrel hydrogen sulfate is as follows: o-chlorobenzene glycine methyl ester tartrate is added into a mixed system of water and toluene, the pH value of the mixed system is adjusted by sodium carbonate solution to obtain a solution of methyl ester and toluene (with a + chiral positive-rotation structure), and then the mixed solution reacts with dipotassium hydrogen phosphate and intermediate p-toluenesulfonic acid thiophene ester under the catalysis of tetrabutylammonium bromide, and then the mixed solution is salified by hydrogen chloride methanol solution to obtain a condensation compound. Wherein the molecular formula of the intermediate p-toluenesulfonic acid thiophene ester is C 13 H 14 O 3 S 2 The molecular weight is: 282.40, the structural formula is as follows:
at present, although pharmacopoeia records exist about the quality control of clopidogrel hydrogen sulfate impurity content, no LCMS detection method has been disclosed on the quality control of impurities (p-toluenesulfonate thiophene ester), the p-toluenesulfonate thiophene ester is taken as a key intermediate, a p-toluenesulfonate genotoxicity warning structure is provided, two or three reaction steps are only carried out on the p-toluenesulfonate thiophene ester as far as a finished product clopidogrel hydrogen sulfate, and in order to effectively control the impurities, an analysis method capable of effectively detecting ppm level of p-toluenesulfonate thiophene ester in clopidogrel hydrogen sulfate is established.
Disclosure of Invention
The purpose of the invention is as follows:
because the p-toluenesulfonate has a p-toluenesulfonate genotoxicity warning structure, in order to effectively control p-toluenesulfonate impurities in clopidogrel hydrogen sulfate and investigate the transmission of the impurities, the invention provides an analysis method capable of effectively measuring the ppm level of the p-toluenesulfonate.
The invention provides the following technical scheme:
an analytical method for measuring p-toluenesulfonate thiophene ester is characterized in that a liquid chromatography-mass spectrometer is adopted for detecting and analyzing p-toluenesulfonate thiophene ester, a chromatographic column with octadecylsilane chemically bonded silica as a filler is adopted, an acid aqueous solution is used as a mobile phase A, a lower alkyl alcohol or acetonitrile is used as a mobile phase B, the volume concentration of the mobile phase A is 0.1-0.5%, and the initial volume fraction of the mobile phase B is 25-50%; carrying out gradient elution, wherein the structural formula of the p-toluenesulfonic acid thiophene ester is as follows:
in some preferred embodiments, the volume concentration of mobile phase a is 0.1% to 0.3%; the initial volume fraction of the mobile phase B is 40-50%.
In some more preferred embodiments, the mobile phase a has a volume concentration of 0.1%; the initial volume fraction of mobile phase B was 45%.
In some embodiments, the detector is a mass spectrometry detector in which the scanning mode is a multiple reaction detection scanning mode.
In some embodiments, the mass spectrometry detector instrument parameters are set to:
| parameter name | Condition of parameter |
| Ion source | ESI source |
| |
0～200V |
| Temperature of drying gas | 250～300℃ |
| Dry |
8～12L/min |
| Pressure of atomized gas | 35～45psi |
| Nozzle voltage | 1400～1600V |
| Temperature of sheath gas | 250～300℃ |
| Flow rate of sheath gas | 5～8L/min |
| Capillary voltage | 3500～4500V |
In some preferred embodiments, the mass spectrometer instrument parameters are set to:
| parameter name | Condition of parameter |
| Ion source | Jet-ESI source |
| Electron multiplication voltage | 0V |
| Temperature of drying gas | 250℃ |
| Dry air flow rate | 11L/min |
| Pressure of atomized gas | 40psi |
| Nozzle voltage | 1500V |
| Temperature of sheath gas | 250℃ |
| Velocity of sheath gas | 8L/min |
| Capillary voltage | 4000V |
In some embodiments, the ion detection parameters in the liquid chromatography mass spectrometer are set as:
| p-toluenesulfonic acid thiophene ester | Parent ion | Daughter ions |
| Quantification of | 305m/z | 111m/z |
| Characterization of nature | 305m/z | 195m/z |
In some embodiments, the aqueous acid solution is an aqueous formic acid solution or an aqueous acetic acid solution. In some preferred embodiments, the aqueous acid solution is aqueous formic acid.
In some embodiments, the lower alkyl alcohol is one or more of methanol, ethanol, isopropanol, propanol, or butanol. In some preferred embodiments, the lower alkyl alcohol is methanol.
In some embodiments, the column temperature is from 30 ℃ to 50 ℃. In some preferred embodiments, the column temperature is 40 ℃.
In some embodiments, the detection wavelength is from 200nm to 220nm. In some preferred embodiments, the detection wavelength is 220nm.
In some embodiments, the sample is introduced in an amount of 5 to 10 μ l, the mobile phase flow rate is 0.5 to 0.9ml/min, and the sample plate is temperature controlled at 2 to 8 ℃. In some preferred embodiments, the sample is taken in an amount of 10. Mu.l, the mobile phase flow rate is 0.7ml/min, and the sample plate is temperature controlled to 4 ℃.
In some embodiments, the gradient elution procedure is:
in some preferred embodiments, the gradient elution procedure is:
| elution time (min) | Mobile phase A (%) | Moving phase B (%) |
| 0 | 60～50 | 40～50 |
| 7 | 20～30 | 80～70 |
| 15 | 20～30 | 80～70 |
| 16 | 60～50 | 40～50 |
| 20 | 60～50 | 40～50 |
In some more preferred embodiments, the gradient elution procedure is:
| elution time (min) | Phase A (%) | Phase B (%) |
| 0 | 55 | 45 |
| 7 | 25 | 75 |
| 15 | 25 | 75 |
| 16 | 55 | 45 |
| 20 | 55 | 45 |
In some embodiments, the chromatography column is Agilent ZORBAXplus C18 (4.6X 100mm,3.5 μm).
In some embodiments, the column: the grain diameter of the filler is 2.0-6.0 μm, the length of the chromatographic column is 100-250 mm, and the inner diameter of the chromatographic column is 4.0-5.0 mm. In some preferred embodiments, the packing has a particle size of 3.5 μm, a column length of 100mm, and a column inner diameter of 4.6mm.
Has the advantages that:
the invention provides an analysis method for measuring p-toluenesulfonate, which shows the detection advantage of p-toluenesulfonate on the ppm level, and has extremely high specificity through the detection of parent ions, quantitative ions and qualitative ions by a liquid chromatography-mass spectrometer. The invention provides an accurate, efficient and highly specific detection method for the ppm level determination of the p-toluenesulfonic acid thiophene ester.
Drawings
FIG. 1 shows the basis for the selection of parent ions and quantitative and qualitative ions according to the present invention;
FIG. 2 is a graph showing the detection results of example 1 of the present invention, wherein RT =11.56min is a peak of 111m/z for quantifying ion in thienyl p-toluenesulfonate;
FIG. 3 is a graph showing the detection results of example 1 of the present invention, wherein RT =11.56min is the signal-to-noise ratio response value of p-toluenesulfonate quantification ion 111m/z at the mass concentration of 50 ng/ml;
FIG. 4 is a graph showing the detection results of example 1 of the present invention, wherein RT =11.56min is the peak of 195m/z for thiophenyl tosylate counterion.
Detailed Description
The technical solutions in the embodiments of the present invention will be described in detail below, but the present invention is not limited to the scope of the embodiments. Unless otherwise indicated, reagents, materials and equipment used in the following examples are available under conventional conditions and methods, or by conventional commercial means.
Example 1
The instrument comprises the following steps: agilent 1260-6460 liquid chromatography-mass spectrometry combined instrument
A chromatographic column: agilent ZORBAXplus C18 (4.6X 100mm,3.5 μm)
Mobile phase: phase A: 0.1% aqueous formic acid solution
Phase B: methanol
Detection wavelength: 220nm
Flow rate: 0.7ml/min
Column temperature: 40 deg.C
Sample introduction amount: 10 μ l
Temperature control of a sample tray: 4 deg.C
Elution procedure:
table 1: example 1 gradient elution procedure
| Elution time (min) | Phase A (%) | Phase B (%) |
| 0min | 55 | 45 |
| 7min | 25 | 75 |
| 15min | 25 | 75 |
| 16min | 55 | 45 |
| 20min | 55 | 45 |
By adopting the elution procedure in the table 1, the separation of p-toluenesulfonate thiophene ester impurities and main components can be ensured, and the phenomenon that the impurity easily causes pollution when the excessive concentration enters a liquid chromatography-mass spectrometer is avoided.
When the proportion, the column temperature, the detection wavelength and the flow rate of the mobile phase in the chromatographic condition are slightly changed, for example, the initial volume fraction of the mobile phase B is 40 to 50 percent, even 25 to 50 percent; the column temperature is 35-45 ℃, even 30-50 ℃; the detection wavelength is 200nm-220nm; the flow rate is 0.5ml/min-0.9ml/min, even 0.5ml/min-1.5ml/min, the detection result is not affected.
An ion source: jet-ESI
The ion detection parameters were set as in table 2 below:
table 2: ion detection parameter setting
| Temperature of drying gas (. Degree.C.) | 250 |
| Flow rate of drying gas (L/min) | 11 |
| Atomizing air pressure (psi) | 40 |
| Temperature of sheath gas (. Degree. C.) | 250 |
| Sheath gas flow rate (L/min) | 8 |
| Capillary voltage (V) | ±4000 |
| Nozzle voltage (V) | ±1500 |
Scanning mode: multiple Reaction detection scanning (Multi Reaction Monitoring abbreviated as MRM)
Mass spectrum acquisition time: 3min to 10.1min
Electron multiplication voltage (+): 0V
As shown in FIG. 1, the basis for the selection of parent ions, quantitative ions and qualitative ions is shown. The method comprises the following specific steps: the molecular weight of p-toluenesulfonate thiophene ester is 282, and during mass spectrum first heavy quadrupole mass separation, a molecular weight of sodium addition, namely, the molecular weight is equivalent to +23, and the parent ion parameter is 305m/z. After the second-time quadrupole collision activation, parent ions are fractured, and the fractured daughter ions with parameters of 111m/z and 195m/z can be obtained in the third-time quadrupole mass separation.
The scan parameter settings are as follows in table 3:
table 3: scan parameter setting
Preparing a system applicability solution: taking a proper amount of p-toluenesulfonate reference substance, precisely weighing, placing in a 100ml volumetric flask, adding methanol for dissolving and diluting, transferring 2.0ml into the 100ml volumetric flask, adding methanol for dissolving and diluting, transferring 1.0ml into the 100ml volumetric flask, adding methanol for dissolving and diluting, and preparing a solution with the p-toluenesulfonate content of about 50ng in each 1ml as a system applicability solution.
And (3) detection results: referring to fig. 2, it can be seen that RT =11.56min is a peak of 111m/z for p-toluenesulfonate quantification ion in this example. Referring to fig. 3, it can be seen that the p-toluenesulfonate quantification ion 111m/z in the present embodiment has a signal-to-noise ratio of 1000 or more under the condition of a mass concentration of 50ng/ml, and has an excellent response. Referring to fig. 4, it can be seen that RT =11.56min is the peak of 195m/z for thiophenyl tosylate counterion in this example. From the results of FIGS. 2 to 4, it can be seen that thiophene p-toluenesulfonate has unique specificity under a plurality of conditions of a parent ion 305m/z, a quantitative ion 111m/z and a qualitative ion 195m/z.
It is obvious to the person skilled in the art that the invention is not limited to the details of the exemplary embodiments described above; and that the present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive. The scope of the invention is indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (10)
1. An analysis method for measuring p-toluenesulfonate thiophene is characterized in that a liquid chromatography-mass spectrometer is adopted to detect and analyze the p-toluenesulfonate thiophene, a chromatographic column with octadecylsilane chemically bonded silica as a filler is adopted, an acid aqueous solution is taken as a mobile phase A, a lower alkyl alcohol or acetonitrile is taken as a mobile phase B, the volume concentration of the mobile phase A is 0.1-0.5%, and the initial volume fraction of the mobile phase B is 25-50%; carrying out gradient elution, wherein the structural formula of the p-toluenesulfonate is as follows:
2. the method of claim 1, wherein the detector is a mass spectrometer detector and the scan pattern in the mass spectrometer detector is a multiple reaction detection scan pattern.
3. The analytical method of claim 1, wherein the ion detection parameters in the LC MS are set as follows:
。
4. The analytical method according to claim 1, wherein the aqueous acid solution is an aqueous formic acid solution or an aqueous acetic acid solution; the lower alkyl alcohol is one or more of methanol, ethanol, isopropanol, propanol or butanol.
5. The analytical method according to claim 1, wherein the column temperature is 30 ℃ to 50 ℃.
6. The analytical method of claim 5, wherein the column temperature is 40 ℃.
7. The analytical method according to claim 1, wherein the amount of the sample is 5 to 10 μ l and the flow rate of the mobile phase is 0.5 to 0.9ml/min.
8. The analytical method according to claim 7, wherein the sample volume is 10 μ l and the mobile phase flow rate is 0.7ml/min.
9. The assay of claim 1, wherein the gradient elution procedure is:
。
10. The assay of claim 1, wherein the gradient elution procedure is:
。
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