CN112730683A - Method for detecting epichlorohydrin isomer - Google Patents
Method for detecting epichlorohydrin isomer Download PDFInfo
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- CN112730683A CN112730683A CN202011568845.2A CN202011568845A CN112730683A CN 112730683 A CN112730683 A CN 112730683A CN 202011568845 A CN202011568845 A CN 202011568845A CN 112730683 A CN112730683 A CN 112730683A
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- BRLQWZUYTZBJKN-UHFFFAOYSA-N Epichlorohydrin Chemical class ClCC1CO1 BRLQWZUYTZBJKN-UHFFFAOYSA-N 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 32
- BRLQWZUYTZBJKN-VKHMYHEASA-N (-)-Epichlorohydrin Chemical compound ClC[C@H]1CO1 BRLQWZUYTZBJKN-VKHMYHEASA-N 0.000 claims abstract description 68
- BRLQWZUYTZBJKN-GSVOUGTGSA-N (+)-Epichlorohydrin Chemical compound ClC[C@@H]1CO1 BRLQWZUYTZBJKN-GSVOUGTGSA-N 0.000 claims abstract description 38
- 238000001514 detection method Methods 0.000 claims abstract description 32
- 238000004817 gas chromatography Methods 0.000 claims abstract description 17
- 238000002347 injection Methods 0.000 claims abstract description 8
- 239000007924 injection Substances 0.000 claims abstract description 8
- 239000000126 substance Substances 0.000 claims abstract description 6
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 4
- 229940080345 gamma-cyclodextrin Drugs 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 30
- 239000013558 reference substance Substances 0.000 claims description 21
- 238000004587 chromatography analysis Methods 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 12
- 239000012085 test solution Substances 0.000 claims description 11
- 239000002904 solvent Substances 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- 230000005526 G1 to G0 transition Effects 0.000 claims description 7
- 239000012159 carrier gas Substances 0.000 claims description 6
- 239000012088 reference solution Substances 0.000 claims description 6
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 4
- 239000011248 coating agent Substances 0.000 claims description 4
- 238000000576 coating method Methods 0.000 claims description 4
- 239000001257 hydrogen Substances 0.000 claims description 4
- 229910052739 hydrogen Inorganic materials 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- IPWKIXLWTCNBKN-ZCFIWIBFSA-N (2s)-1-chloro-3-(2-methyl-5-nitroimidazol-1-yl)propan-2-ol Chemical compound CC1=NC=C([N+]([O-])=O)N1C[C@H](O)CCl IPWKIXLWTCNBKN-ZCFIWIBFSA-N 0.000 abstract description 8
- 238000001212 derivatisation Methods 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000012795 verification Methods 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 2
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 54
- 239000000523 sample Substances 0.000 description 19
- 239000011259 mixed solution Substances 0.000 description 16
- 239000007789 gas Substances 0.000 description 14
- 238000000926 separation method Methods 0.000 description 12
- 238000011084 recovery Methods 0.000 description 11
- 230000000052 comparative effect Effects 0.000 description 9
- 238000005303 weighing Methods 0.000 description 8
- 239000012488 sample solution Substances 0.000 description 6
- 230000002349 favourable effect Effects 0.000 description 4
- ICKWICRCANNIBI-UHFFFAOYSA-N 2,4-di-tert-butylphenol Chemical compound CC(C)(C)C1=CC=C(O)C(C(C)(C)C)=C1 ICKWICRCANNIBI-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- IPWKIXLWTCNBKN-UHFFFAOYSA-N Madelen Chemical compound CC1=NC=C([N+]([O-])=O)N1CC(O)CCl IPWKIXLWTCNBKN-UHFFFAOYSA-N 0.000 description 1
- 208000022506 anaerobic bacteria infectious disease Diseases 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000842 anti-protozoal effect Effects 0.000 description 1
- 239000013064 chemical raw material Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 150000004957 nitroimidazoles Chemical class 0.000 description 1
- 229960002313 ornidazole Drugs 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
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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
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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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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Abstract
The invention relates to the technical field of chemical analysis, and particularly discloses a method for detecting an epichlorohydrin isomer. The method adopts a capillary column with a fixed phase of 2, 3-di-O-acetyl-6-0-TBDMS-gamma-cyclodextrin embedded with a middle polar phase as a detection chromatographic column, and sets the column temperature to be operated at a constant temperature of 80-90 ℃, the injection port temperature is 245-255 ℃, the detector temperature is 245-255 ℃, and the accurate quantitative detection of R-epichlorohydrin in an S-epichlorohydrin sample can be realized by gas chromatography without derivatization treatment of S-epichlorohydrin. Proved by methodological verification, the epichlorohydrin isomer detection method provided by the invention has the advantages of good accuracy, repeatability and stability, high detection sensitivity, and simple and convenient operation, can be used for qualitatively and quantitatively analyzing R-epichlorohydrin in S-epichlorohydrin, and is further beneficial to realizing the control of purity and quality safety of the levoornidazole product.
Description
Technical Field
The invention relates to the technical field of chemical analysis, in particular to a method for detecting an epichlorohydrin isomer.
Background
Epichlorohydrin is also known as epichlorohydrin, is an important organic chemical raw material and fine chemical product, is active in chemical property and chiral molecules, can react with other compounds through functional group conversion reaction to prepare a series of corresponding chiral drugs with different structures, and has wide application.
The chemical name of the levoornidazole is (S) - (-) -1- (3-chloro-2-hydroxypropyl) -2-methyl-5-nitroimidazole, which is a 3 rd generation nitroimidazole derivative, has the characteristics of strong antibacterial capability, high antiprotozoal activity, long half-life period, good tissue permeability and the like, and is mainly used for various diseases caused by anaerobic bacterial infection clinically. S-epichlorohydrin is an important starting material in the synthesis of the ornidazole. However, during the synthesis of epichlorohydrin, the isomer (R-epichlorohydrin) inevitably occurs. The existence of the R-epichlorohydrin isomer can directly influence the preparation of the levoornidazole, so that the purity of the product is low, and the safety risk of the levoornidazole product is increased. In the prior art, the method for detecting the epichlorohydrin isomer needs to adopt a chiral derivatization reagent to perform derivatization treatment on the epichlorohydrin, and the operation steps are complicated. Therefore, it is necessary to develop a simple method for detecting epichlorohydrin isomers to improve the detection effect, thereby effectively controlling the purity and quality safety of the levoornidazole product.
Disclosure of Invention
The invention provides a method for detecting an epichlorohydrin isomer, aiming at the technical problems that the method for detecting the epichlorohydrin isomer in the prior art needs derivatization treatment and is complex to operate.
In order to solve the technical problems, the technical scheme provided by the invention is as follows:
a method for detecting an epichlorohydrin isomer is a gas chromatography method, and comprises the following steps:
(1) preparing a test solution and a reference solution:
preparing a reference substance solution: preparing a reference substance solution from an R-epichlorohydrin reference substance by using a solvent;
preparing a test solution: preparing a sample of S-epichlorohydrin into a test solution by using a solvent;
(2) taking a reference substance solution and a test substance solution, and detecting according to the following gas chromatography conditions:
a detector: a hydrogen flame ionization detector;
a chromatographic column: the stationary phase is a capillary column of 2, 3-di-O-acetyl-6-0-TBDMS-gamma-cyclodextrin embedded in a medium polar phase;
column temperature: running at constant temperature of 80-90 ℃;
sample inlet temperature: 245 ℃ and 255 ℃;
detector temperature: 245 ℃ and 255 ℃;
the split ratio is as follows: 95-105: 1;
sample introduction volume: 0.05-0.15 μ L.
Compared with the prior art, the method for detecting the isomer of the epichlorohydrin provided by the invention adopts the capillary column with the stationary phase of the 2, 3-di-O-acetyl-6-0-TBDMS-gamma-cyclodextrin embedded in the middle polar phase as the detection chromatographic column, and can realize the quantitative detection of the R-epichlorohydrin in the S-epichlorohydrin sample by matching with the specific gas chromatographic condition without performing derivatization treatment on the S-epichlorohydrin. Proved by methodological verification, the epichlorohydrin isomer detection method provided by the invention has the advantages of good accuracy, repeatability and stability, high detection sensitivity, and simple and convenient operation, can be used for qualitatively and quantitatively analyzing R-epichlorohydrin in S-epichlorohydrin, and is further beneficial to realizing the control of purity and quality safety of the levoornidazole product.
Preferably, the column length of the chromatographic column is 15-60m, the inner diameter is 0.25-0.53mm, and the thickness of the stationary phase coating liquid film is 0.25-0.53 μm.
Further preferably, the chromatographic column has a column length of 30m, an inner diameter of 0.25mm, and a stationary phase coating liquid film thickness of 0.25 μm.
Further preferably, the column is a Gamma DEXTM 225, 30 m.times.0.25 mm.times.0.25 μm.
The preferable chromatographic column is favorable for realizing the separation of the S-epichlorohydrin and the R-epichlorohydrin, thereby being favorable for accurately and quantitatively detecting the R-epichlorohydrin in the S-epichlorohydrin by gas chromatography.
Preferably, the column temperature is 80 ℃, the injection port temperature is 250 ℃, and the detector temperature is 250 ℃.
Preferably, the carrier gas is nitrogen and the flow rate is 0.5-0.7 mL/min.
More preferably, the carrier gas flow rate is 0.6 mL/min.
Preferably, the split ratio is 100: 1.
Preferably, the injection volume is 0.1. mu.L.
Preferably, the solvent is anhydrous methanol.
Preferably, the concentration of the R-epichlorohydrin in the control solution is 250 mug/mL, and the concentration of the test solution is 50 mg/mL.
The optimized gas chromatography condition is favorable for improving the separation degree of S-epichlorohydrin and R-epichlorohydrin, thereby being favorable for accurately quantifying the R-epichlorohydrin.
Through the research and verification of methodologies such as accuracy, repeatability and sensitivity, the gas chromatography detection method provided by the invention finds that the method has better sensitivity, accuracy and repeatability, can realize quantitative detection on the epichlorohydrin isomer by a simpler method, further provides reliable guarantee for better controlling the quality of the levoornidazole product, and has wide application prospect.
Drawings
FIG. 1 is a gas chromatogram of a mixed solution detected using LIPODEX chiral capillary chromatography column in example 1 of the present invention;
FIG. 2 is a gas chromatogram of the mixed solution detected by using a CYCLOSIL-B chiral capillary chromatographic column in example 1 of the present invention;
FIG. 3 is a gas chromatogram of the mixed solution detected by using a CYCLODEX-B chiral capillary chromatographic column in example 1 of the present invention;
FIG. 4 is a gas chromatogram of a mixed solution detected by an Astec CHIRALDEX B-DM chiral capillary chromatographic column in example 1 of the present invention;
FIG. 5 is a gas chromatogram of a mixed solution detected by an Astec CHIRALDEX G-TA chiral capillary chromatographic column in example 1 of the present invention;
FIG. 6 is a gas chromatogram of a mixed solution detected by using a chiral capillary chromatographic column of Gamma DEXTM 225 in example 1 of the present invention;
FIG. 7 is a gas chromatogram of a methanol solvent in example 2 of the present invention;
FIG. 8 is a gas chromatogram of a comparison solution of R-epichlorohydrin in example 2 of the present invention;
FIG. 9 is a gas chromatogram of the mixed solution in example 2 of the present invention;
FIG. 10 is a gas chromatogram of the mixed solution in comparative example 1 of the present invention;
FIG. 11 is a gas chromatogram of the mixed solution in comparative example 2 of the present invention;
FIG. 12 is a gas chromatogram of the mixed solution in comparative example 3 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
1.1 preparation of solution:
preparing a reference substance solution: taking a proper amount of R-epichlorohydrin reference substance, precisely weighing, and preparing a reference substance solution with the concentration of 250 mug/mL by using methanol.
Preparing a test solution: taking a proper amount of S-epichlorohydrin sample, precisely weighing, and preparing a sample solution with the concentration of 50mg/mL by using methanol.
Mixing the solution: taking a proper amount of each of an S-epichlorohydrin sample and an R-epichlorohydrin reference substance, and preparing a mixed solution with the concentration of the R-epichlorohydrin of 250 mu g/mL and the concentration of the S-epichlorohydrin of 50mg/mL by using methanol.
1.2 gas chromatography conditions:
a detector: a hydrogen flame ionization detector;
column temperature: the operation is carried out at a constant temperature of 85 ℃;
sample inlet temperature: 250 ℃;
detector temperature: 250 ℃;
carrier gas: high purity nitrogen (purity 99.99%), flow rate 0.6 mL/min;
the split ratio is as follows: 100: 1;
sample introduction volume: 0.1. mu.L.
1.3 selection of chromatographic column:
in the early stage of the experiment, the inventor tries to analyze and detect R-epichlorohydrin in S-epichlorohydrin by various methods, and selects various chromatographic columns to detect the mixed solution according to the methods.
Detection was carried out using a chiral capillary chromatography column of LIPODEX (25 m. times.0.25 mm. times.0.25 μm) under the chromatography conditions of item 1.2, and the results are shown in FIG. 1. As can be seen from the figure, the peak shape of the R-epichlorohydrin is poor, and the R-epichlorohydrin can not be accurately quantified.
The detection was carried out by using a chiral capillary chromatography column of CYCLOSIL-B (30 m. times.0.32 mm. times.0.25 μm) under the chromatography conditions of item 1.2, and the results are shown in FIG. 2. As can be seen from the figure, chromatographic peaks of the S-epichlorohydrin and the R-epichlorohydrin are completely coincident, peak positions of the S-epichlorohydrin and the R-epichlorohydrin are about 9.8min, and effective separation of the R-epichlorohydrin in the S-epichlorohydrin cannot be realized.
The detection was carried out by using a chiral capillary chromatography column of CYCLODEX-B (30 m. times.0.32 mm. times.0.25 μm) under the chromatography conditions of item 1.2, and the results are shown in FIG. 3. As can be seen from the figure, chromatographic peaks of S-epichlorohydrin and R-epichlorohydrin are completely coincident, peak positions of the S-epichlorohydrin and the R-epichlorohydrin are about 10min, and effective separation of the R-epichlorohydrin in the S-epichlorohydrin cannot be realized.
The detection was carried out by using a chiral capillary chromatography column (30 m. times.0.25 mm. times.0.12 μm) of Astec CHIRALDEX B-DM under the chromatography conditions of item 1.2, the results of which are shown in FIG. 4. As can be seen from the figure, chromatographic peaks of the S-epichlorohydrin and the R-epichlorohydrin are completely coincident, peak positions of the S-epichlorohydrin and the R-epichlorohydrin are both about 8.5min, and effective separation of the R-epichlorohydrin in the S-epichlorohydrin cannot be realized.
The detection was carried out by using a chiral capillary chromatography column (30 m. times.0.25 mm. times.0.12 μm) of Astec CHIRALDEX G-TA under the chromatography conditions of item 1.2, and the results are shown in FIG. 5. As can be seen from the figure, chromatographic peaks of S-epichlorohydrin and R-epichlorohydrin can not be effectively separated, and accurate quantification of R-epichlorohydrin can not be realized.
The inventors surprisingly found that effective separation of the chromatographic peaks of S-epichlorohydrin and R-epichlorohydrin can be achieved using a chiral capillary chromatographic column of type Gamma dex tm 225(30m x 0.25mm x 0.25 μm). The results of the detection according to the chromatographic conditions under item 1.2 are shown in FIG. 6. As can be seen from the figure, chromatographic peaks of S-epichlorohydrin and R-epichlorohydrin can be effectively separated, the peak-off time of S-epichlorohydrin is 7.838min, the peak-off time of R-epichlorohydrin is 8.398min (3.340min is a methanol solvent peak), the separation degree of R-epichlorohydrin and S-epichlorohydrin is 3.41, the separation degree and the peak shape are good, and the detection requirement is met. Therefore, this column was selected as the column for detecting epichlorohydrin isomers.
Example 2
And (3) verification of methodology:
2.1 specificity
The methanol solvent, the control solution and the mixed solution are detected according to the gas chromatography conditions, and chromatograms are recorded, and the results are shown in fig. 7-9, wherein fig. 7 is a chromatogram of the methanol solvent, fig. 8 is a chromatogram of the control solution, and fig. 9 is a chromatogram of the mixed solution.
As can be seen from FIG. 7, the methanol peak time was 3.332 min. As can be seen from FIG. 8, the peak time of R-epichlorohydrin was 8.418 min. As can be seen from FIG. 9, the peak time of S-epichlorohydrin is 7.832min, the peak time of R-epichlorohydrin is 8.387min, and the separation degree of the two is 3.42. The method is proved to have good specificity.
2.2 detection and quantitation limits
Respectively taking a proper amount of R-epichlorohydrin, precisely weighing, preparing a reference substance solution with the concentration of about 250 mu g/mL by using methanol, precisely measuring the reference substance solution, performing serial dilution, respectively injecting into a gas chromatograph, and recording a chromatogram. When the chromatographic peak height of the R-epichlorohydrin is about 3 times of the baseline noise, the chromatographic peak height is the detection limit; the results are shown in table 1 for quantitation when the R-epichlorohydrin chromatographic peak height is about 10 times the baseline noise. 6 parts of a reference substance solution is prepared according to the quantitative limit concentration, and the detection is carried out according to the gas chromatography conditions, and the results are shown in Table 2.
TABLE 1 detection limit and quantitation limit test results
TABLE 2 quantitative limit repeatability test results
| Sample (I) | 1 | 2 | 3 | 4 | 5 | 6 | RSD/％ |
| Peak area of R-epichlorohydrin | 1.00 | 1.26 | 1.21 | 1.13 | 1.19 | 1.29 | 8.84 |
The test result shows that the detection limit concentration of the R-epichlorohydrin is 25.49 mu g/mL, the quantitative limit concentrations are 84.97 mu g/mL respectively, the quantitative limit solution is repeatedly injected for 6 times, and the RSD of the R-epichlorohydrin peak area is only 8.84%, which indicates that the method has high sensitivity and good quantitative limit repeatability.
2.2 Linear Range
An appropriate amount of R-epichlorohydrin reference substance is precisely weighed, and a series of linear solutions with concentration are respectively prepared by methanol. Respectively and precisely measuring 0.1 μ L, performing sample injection detection according to the gas chromatography conditions, recording chromatogram, measuring peak area, and taking concentration (μ g/mL) as abscissa and peak area as ordinate to make linear regression curve, wherein the result is shown in Table 3.
TABLE 3R-Epichlorohydrin linearity test results
The results show that the R-epichlorohydrin has good linearity in the concentration range of 84.97-510.44 mu g/ml, and the linear correlation coefficient R is 0.9978.
2.3 solution stability
Accurately weighing a proper amount of R-epichlorohydrin, preparing a reference solution with methanol to be about 250 mu g/mL, carrying out sample injection detection for 0h, 2h, 4h, 8h and 12h respectively according to the gas chromatography conditions, recording a chromatogram, and calculating RSD according to the peak area of the R-epichlorohydrin, wherein the results are shown in Table 4.
TABLE 4 solution stability test results
| Time | 0h | 2h | 4h | 8h | 12h | RSD％ |
| Peak area of R-epichlorohydrin | 3.77 | 3.70 | 3.64 | 3.72 | 3.73 | 1.28 |
The test result shows that the R-epichlorohydrin reference solution is placed at room temperature for 12 hours, the RSD of the peak area is 1.28%, and the reference solution has good stability.
2.4 repeatability
6 parts of S-epichlorohydrin sample is taken, and is respectively prepared into a sample solution with the concentration of 50mg/mL by using methanol, and the detection is carried out according to the gas chromatography conditions, and the results are shown in Table 5.
TABLE 5 results of the repeatability tests
| Repeatability of | 1 | 2 | 3 | 4 | 5 | 6 |
| Content of R-epichlorohydrin (%) | Not detected out | Not detected out | Not detected out | Not detected out | Not detected out | Not detected out |
The test result shows that no R-epichlorohydrin is detected in 6 parts of test solution, and the method has good repeatability.
2.5 recovery
Taking a proper amount of R-epichlorohydrin reference substance, precisely weighing, and preparing a stock solution containing about 2500 mu g of R-epichlorohydrin in each 1mL by using methanol; taking 0.5g of S-epichlorohydrin sample, precisely weighing, placing in a 10mL volumetric flask, precisely adding a proper amount of stock solution respectively, and performing constant volume with methanol to prepare solutions containing low, medium and high concentrations of R-epichlorohydrin, wherein 3 parts of each concentration is prepared in parallel to serve as a recovery rate sample solution.
The recovery rates of the 9 recovery rate solutions were measured under the conditions of the gas chromatography, and chromatograms were recorded to calculate the recovery rates, and the results are shown in table 6.
TABLE 6 recovery test results
The test result shows that the average recovery rate of the R-epichlorohydrin in 6 parts of test sample solution is 103.58%, the average recovery rate of the RSD is 1.86%, the average recovery rate of the 2, 4-di-tert-butylphenol is 93.25%, the average recovery rate of the RSD is 1.19%, and the method has good recovery rate.
Example 3
The present embodiment provides a method for detecting an epichlorohydrin isomer:
preparing a reference substance solution: taking a proper amount of R-epichlorohydrin reference substance, precisely weighing, and preparing a reference substance solution with the concentration of 250 mug/mL by using methanol.
Preparing a test solution: taking a proper amount of S-epichlorohydrin sample, precisely weighing, and preparing a sample solution with the concentration of 50mg/mL by using methanol.
Mixing the solution: taking a proper amount of each of an S-epichlorohydrin sample and an R-epichlorohydrin reference substance, and preparing a mixed solution with the concentration of the R-epichlorohydrin of 250 mu g/mL and the concentration of the S-epichlorohydrin of 50mg/mL by using methanol.
Gas chromatography conditions:
a detector: a hydrogen flame ionization detector;
a chromatographic column: gamma DEXTM 225(30m 0.25mm 0.25 μm);
column temperature: the operation is carried out at a constant temperature of 85 ℃;
sample inlet temperature: 250 ℃;
detector temperature: 250 ℃;
carrier gas: high purity nitrogen (purity 99.99%), flow rate 0.6 mL/min;
the split ratio is as follows: 100: 1;
sample introduction volume: 0.1. mu.L.
And (3) sampling and detecting the mixed solution according to the gas chromatography conditions, sampling and detecting the reference solution and the sample solution when the separation degree of the S-epichlorohydrin and the R-epichlorohydrin is qualified, and indicating that the R-epichlorohydrin is not detected in the sample.
The other chromatographic conditions defined by the invention can also achieve the purpose of qualified separation degree of S-epichlorohydrin and R-epichlorohydrin, and can also achieve the detection effect basically equivalent to that of the embodiment 3.
Comparative example 1
The comparative example provides a method for detecting an epichlorohydrin isomer, which is completely the same as that in example 3, except that the chromatographic column is operated at a constant temperature of 50 ℃ for 30min under chromatographic conditions, and a chromatogram is shown in FIG. 10. As can be seen from the figure, the R-epichlorohydrin peak pattern is poor, and the R-epichlorohydrin peak pattern cannot be accurately quantified.
Comparative example 2
This comparative example provides a method for detecting an epichlorohydrin isomer, which is exactly the same as in example 3 except that the separation ratio under chromatographic conditions was 50:1 and the chromatogram is shown in FIG. 11. As can be seen from the figure, the R-epichlorohydrin peak pattern is poor, and the R-epichlorohydrin peak pattern cannot be accurately quantified.
Comparative example 3
This comparative example provides a method for detecting an epichlorohydrin isomer, which is the same as that of example 3 except that the sample injection volume is 2. mu.L under chromatographic conditions, and the chromatogram is shown in FIG. 12. As can be seen from the figure, the R-epichlorohydrin peak pattern is poor, and the R-epichlorohydrin peak pattern cannot be accurately quantified.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. The method for detecting the epichlorohydrin isomer is characterized by being a gas chromatography method and comprising the following steps:
(1) preparing a test solution and a reference solution:
preparing a reference substance solution: preparing a reference substance solution from an R-epichlorohydrin reference substance by using a solvent;
preparing a test solution: preparing a sample of S-epichlorohydrin into a test solution by using a solvent;
(2) taking a reference substance solution and a test substance solution, and detecting according to the following gas chromatography conditions:
a detector: a hydrogen flame ionization detector;
a chromatographic column: the stationary phase is a capillary column of 2, 3-di-O-acetyl-6-0-TBDMS-gamma-cyclodextrin embedded in a medium polar phase;
column temperature: running at constant temperature of 80-90 ℃;
sample inlet temperature: 245 ℃ and 255 ℃;
detector temperature: 245 ℃ and 255 ℃;
the split ratio is as follows: 95-105: 1;
sample introduction volume: 0.05-0.15 μ L.
2. The epichlorohydrin isomer detection method according to claim 1, wherein a column length of the chromatographic column is 15 to 60m, an inner diameter is 0.25 to 0.53mm, and a liquid film thickness of the stationary phase coating is 0.25 to 0.53 μm.
3. The epichlorohydrin isomer detection method according to claim 2, wherein the column length of the chromatographic column is 30m, the inner diameter is 0.25mm, and the thickness of the stationary phase coating liquid film is 0.25 μm.
4. The epichlorohydrin isomer detection method of claim 2, wherein the chromatography column is Gamma dex tm 225, 30m x 0.25mm x 0.25 μm.
5. The method for detecting an epichlorohydrin isomer according to claim 1, wherein the column temperature is 80 ℃, the injection port temperature is 250 ℃, and the detector temperature is 250 ℃.
6. The epichlorohydrin isomer detection method of claim 1, wherein the carrier gas is nitrogen and the flow rate is 0.5-0.7 mL/min.
7. The epichlorohydrin isomer detection method of claim 6, wherein the carrier gas flow rate is 0.6 mL/min.
8. The method for detecting an epichlorohydrin isomer according to claim 1, wherein the split ratio is 100: 1.
9. The method for detecting an epichlorohydrin isomer according to claim 1, wherein the injection volume is 0.1 μ L.
10. The method for detecting an epichlorohydrin isomer according to claim 1, wherein the concentration of R-epichlorohydrin in the control solution is 250 μ g/mL, and the concentration of the test solution is 50 mg/mL.
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