CN115266981B - Method for detecting deuteration rate of substance by GC-MS - Google Patents
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Abstract
The invention relates to a method for detecting the deuteration rate of a substance by using GC-MS, preparing a certain amount of sample before deuteration and sample solution after deuteration to be detected, and detecting by using the GC-MS; integrating main peaks of the samples before and after deuteration after blank deduction, and extracting ions, response values and abundance percentages of the main peaks; and (3) extracting main peak ions before and after deuteration, response values and abundance percentage data of the obtained samples, and calculating each deuteration rate of the samples. Compared with the traditional nuclear magnetic method for evaluating the deuteration rate, the method has low requirements on instruments and equipment, can separate impurities by adjusting analysis parameters at a GC end, has no special requirements on the purity of an analysis sample, is simple in sample pretreatment, can evaluate the deuterium quantity ratio contained in the compound, and can correct a detection instrument by using the sample before deuteration, deduct isotope interference and the like, so that the analysis result is more accurate and reliable.
Description
Technical Field
The invention belongs to the field of mass analysis, and particularly relates to a method for detecting the deuteration rate of a substance by using GC-MS.
Background
Deuterium (D), also known as deuterium, is a stable isotope of hydrogen (H), carries neutrons deficient in hydrogen, has a mass 2 times that of hydrogen, has similar chemical properties as hydrogen, but has a carbon-deuterium bond shorter than a carbon-hydrogen bond, and a low vibration stretching frequency of the carbon-deuterium bond makes the carbon-deuterium bond lower in ground state energy than the carbon-hydrogen bond, and is more stable in structure, and in view of these characteristics, in the pharmaceutical production, a deuteration method is often used to improve pharmacokinetic characteristics and metabolic spectrum of the drug without losing pharmaceutical activity, so as to achieve the purposes of reducing the administration frequency or administration dose, reducing adverse effects of the drug, and the like.
The organic electroluminescent (OLED) display technology has the advantages of high resolution, quick response, wide viewing angle, low power consumption, colorization, flexible screen, wide requirement on environmental temperature and the like, the application proportion in the fields of electronic and electric products, medical treatment, traffic, business and the like is improved year by year, but the defects of poor material stability, short service life and the like exist, the application of an organic light-emitting diode is limited, and in order to improve the stability of an OLED material, the deuteration method for the organic light-emitting diode material is a practical and feasible method. For analysis of deuteration rate, most of the literature relates to analysis of some deuteration solvents such as heavy water, and the analysis of deuteration rate of organic compounds is freshly reported, and patent CN112946092A also discloses an evaluation method of deuteration purity of deuteration organic luminescent materials, which combines nuclear magnetism, mass spectrum and ultraviolet; from the published data, it is found that the deuteration rate is basically used for nuclear magnetism and even quantitative nuclear magnetism, the fund investment on the instrument and the equipment is large, in addition, the nuclear magnetism is used for evaluating the deuteration rate, in order to reduce the purity requirement of impurity interference analysis on the sample, expensive deuteration reagents and the like are needed, in addition, in the deuteration process of the sample, the number of deuterium can be different among molecules of the deuterated sample, for example, some molecules only have one deuterium, some molecules have two or three deuterium, even some deuterium does not have the same value, and no detection method has been reported on how to evaluate the quantity of the last deuterium in the sample.
Disclosure of Invention
Aiming at the requirements in the prior art, the invention provides a method for detecting the deuteration rate of a substance by using GC-MS, and the deuteration rate analysis method has low requirements on equipment, is simple and convenient to operate, has scientific and reliable analysis results, and has important guiding significance on production activities.
The invention is realized by the following technical scheme:
a method for assaying deuteration of a substance by GC-MS comprising the steps of:
1) Weighing a certain amount of pre-deuterated sample and a certain amount of post-deuterated sample to be detected, adding a proper amount of solvent to completely dissolve the sample, and preparing a sample blank;
2) Taking a proper amount of the solution in the step 1, passing through an organic filter membrane, and bottling and testing the solution as an upper sample solution;
3) Detecting the sample solution in the step 2 by using GC-MS;
4) Integrating main peaks of the samples before and after deuteration after deducting blank according to the result obtained by GC-MS detection in the step 3, and extracting ions, response values and abundance percentages of the main peaks;
5) And (3) extracting main peak ions before and after deuteration, response values and abundance percentage data in the step (4), and calculating the deuteration rate of the sample.
The structure of the deuterated pre-sample is shown as formula I, and the molecular formula is C 22 H 26 The theoretical m/z is 290.20,291.21 and 292.21 respectively; the abundance percentage is 100.0%,24.1% and 2.8% respectively;
the method of the step 1) is to respectively weigh about 10mg of a pre-deuterated sample and a post-deuterated sample to be detected, and add a solvent with corresponding volume to dissolve the pre-deuterated sample and the post-deuterated sample to be detected to prepare 1mg/mL of on-machine solution; the sample before deuteration is a sample before deuteration of the product; the blank sample is a blank solution prepared by the same steps as the sample except that no sample is added.
The solvent is dichloromethane and tetrahydrofuran, the solution is bottled in a 20mL brown glass bottle, and the dissolution is shaking or ultrasonic until the sample is completely dissolved.
The solvent is dichloromethane.
The organic filter membrane is bottled by a nylon needle filter with the size of 0.45 mu m, and the bottle is a brown sample bottle for chromatographic detection with the size of 2 mL.
The GC-MS detection method in the step 3) comprises the following conditions: gasification chamber temperature: 320 ℃; column flow rate: 1.2ml/min; column temperature: maintaining at 100deg.C for 1min, heating to 250deg.C at 10deg.C/min, and maintaining for 1min; heating to 320 ℃ at 20 ℃/min and keeping for 9.5min; split ratio 30:1, a step of; chromatographic column: agilent HP-5ms UI 30m x 250 μm x 0.25 μm; MS interface temperature: 330 ℃; intuvo flow path: 320 ℃; the sample injection amount is as follows: 1 microliter. Ion source: EI; electron energy: 70eV, MS ion source 230 ℃; MS quadrupole rod 150 ℃; solvent delay time is 3min, the solvent A (methylene dichloride) and the solvent B (methanol) are respectively used for 5 times before and after sample injection, the sample to be injected is used for 3 times before sample injection, then suction is carried out for 4 times, the bubbles of the sample injection needle are removed, and carrier gas is generated: high purity helium, ion scan range: 35-350.
The retention time of the main peak in the step 4) is 19.5+/-0.5 min.
The result of the ion group peak of the main peak molecule before deuteration in the step 4) is recorded as M, and the result of the isotopic abundance percentage of the ion group of the main peak molecule before deuteration is recorded as 100:B:C.
The method for calculating the deuteration rate of the sample comprises the following 4 steps:
(1) According to the molecular ion group peak M of the sample before deuteration, the molecular ion group peak M/z of non-deuteration is obtained as follows: m+0; the ionic group peak m/z of 1 deuterated (upper 1 deuterium) molecule is: m+1; the 2 deuterated (upper 2 deuterated) molecular ionic groups peak m/z is: m+2;3 deuterated (upper 3 deuterated) molecular ionic groups peak m/z is: m+3; the ion group peak m/z of 4 deuterated (4 deuterium) molecules is: m+4; the 5 deuterated (upper 5 deuterated) molecular ion groups peak m/z is: m+5; … …;
(2) And calculating the response value of each deuterated base peak possibly existing in the deuterated sample, wherein the calculation formula is as follows: 0 deuterated group peak M has response value A 0 The method comprises the steps of carrying out a first treatment on the surface of the 1 deuterated group peak m/z response value A 1 Response value-a=m+1 0 * B/100;2 deuterated group peak m/z response value A 2 Response-a =m+2 0 *C/100-A 1 * B/100;3 deuterated group peak m/z response value A 3 Response-a =m+3 1 *C/100-A 2 * B/100;4 deuterated group peak m/z response value A 4 Response-a =m+4 2 *C/100-A 3 * B/100;5 deuterated group peak m/z response value A 5 =m+5 response-a 3 *C/100-A 4 * B/100 … …; until a negative value occurs in the result;
(3) The deuterated non-negative base peak response values are then summed, i.e., A 0 +A 1 +A 2 +A 3 +A 4 +A 5 +……＝K；
(4) After calculation of deuterationRatio of deuteration of each sample: non-deuteration=a 0 K is 100%,1 deuteration=a 1 K is 100%,2 deuteration=a 2 K is 100%,3 deuteration=a 3 K is 100%,4 deuteration=a 4 K is 100%,5 deuteration=a 5 /K*100％……。
Compared with the traditional nuclear magnetic method for evaluating the deuteration rate, the method has low requirements on instruments and equipment, can separate impurities by adjusting analysis parameters at a GC end, has no special requirements on the purity of an analysis sample, is simple in sample pretreatment, can evaluate the deuterium quantity ratio contained in the compound, and can correct a detection instrument by using the sample before deuteration, deduct isotope interference and the like, so that the analysis result is more accurate and reliable.
Drawings
FIG. 1 is the main peak molecular ion of the pre-deuterated sample under method 1.3.1;
FIG. 2 is the main peak molecular ion of deuterated sample 1 under the method of 1.3.1;
FIG. 3 is the main peak molecular ion of deuterated sample 2 under the method of 1.3.1;
FIG. 4 is a graph comparing HNMR of the compound before deuteration and after 2 deuteration;
FIG. 5 is a structural change of the compounds before deuteration and after 2 deuteration;
FIG. 6 is a TIC mass spectrum under the 1.3.1 method after 2 deuteration.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1
The invention provides a method for detecting the deuteration rate of a substance by using GC-MS, which is completed by the following specific steps:
the sample has deuterated structure, theoretical m/z and abundance percentage as shown in formula I and molecular formula C 22 H 26 The theoretical m/z and abundance percentages are 290.20 (100.0%), 291.21 (24.1%), 292.21 (2.8%).
1.1 laboratory instruments and reagents:
agilent gas chromatograph-mass spectrometer Intuvo 9000-5977B; a mertretolide one ten thousandth analytical balance; 0.45 μm organic filter membrane; HPLC grade methanol, dichloromethane; other laboratory glassware such as 2mL sample bottles.
1.2 sample pretreatment:
respectively weighing 10mg of each of the sample 1 and the sample 2 before deuteration and after deuteration by different methods in a 20mL brown glass bottle, adding 10mL of dichloromethane, slightly shaking or performing ultrasonic treatment until the mixture is fully dissolved, taking a proper amount of organic filter membrane as a sample solution, bottling the sample solution for measurement, and preparing a sample blank at the same time, wherein the concentration of the sample is 1 mg/mL.
1.3GC-MS method validation
1.3.1 the sample solution and blank obtained in step 1.2 were examined using the following conditions, wherein the GC-MS method is: gasification chamber temperature: 320 ℃; column flow rate: 1.2ml/min; column temperature: maintaining at 100deg.C for 1min, heating to 250deg.C at 10deg.C/min, and maintaining for 1min; heating to 320 ℃ at 20 ℃/min and keeping for 9.5min; split ratio 30:1, a step of; chromatographic column: agilent HP-5ms UI 30m x 250 μm x 0.25 μm; MS interface temperature: 330 ℃; intuvo flow path: 320 ℃; the sample injection amount is as follows: 1 microliter. Ion source: EI; electron energy: 70eV, MS ion source 230 ℃; MS quadrupole rod 150 ℃; solvent delay time is 3min, the solvent A (methylene dichloride) and the solvent B (methanol) are respectively used for 5 times before and after sample injection, the sample to be injected is used for 3 times before sample injection, then suction is carried out for 4 times, the bubbles of the sample injection needle are removed, and carrier gas is generated: high purity helium, ion scan range: 35-350, total analysis time is 30.0min, and main peak retention time is about 19.5+/-0.5 min; the main peak ion result before deuteration after blank subtraction is shown in figure 1, the main peak ion result of deuterated sample 1 is shown in figure 2, and the main peak ion result of deuterated sample 2 is shown in figure 3.
1.3.2 As can be seen from FIG. 1, the deuterated precursor molecule ion group peak M is 290.2.
1.3.3 As can be seen from FIG. 1, the percent deuterated precursor molecule ion abundance results were 100:B:C=100:25.12:2.82.
1.3.4 molecular ion group peaks M of pre-deuterated samples according to FIG. 1: 290.2, the peak m/z of non-deuterated molecular ion groups which may exist in the deuterated sample is calculated as: 290.2+0=290.2; the ionic group peak m/z of 1 deuterated (upper 1 deuterium) molecule is: 290.2+1=291.2; the 2 deuterated (upper 2 deuterated) molecular ionic groups peak m/z is: 290.2+2= 292.2;3 deuterated (upper 3 deuterated) molecular ionic groups peak m/z is: 290.2+3= 293.2; the ion group peak m/z of 4 deuterated (4 deuterium) molecules is: 290.2+4= 294.2; the 5 deuterated (upper 5 deuterated) molecular ion groups peak m/z is: 290.2+5=295.2; … … from the results of fig. 2, the sample may have non-deuterated, 1 deuterated, 2 deuterated, 3 deuterated, 4 deuterated, and 5 deuterated molecules present.
1.3.5 from the results of fig. 2, the corresponding values of the deuterated base peaks that may be present in deuterated sample 1 were calculated according to the following steps:
1.3.5.10 deuterated group peak m/z response value A 0 ＝5177.13；
1.3.5.2 1 deuterated (upper 1 deuterium) base peak m/z response value A 1 Response value=291.2
-A 0 *B/100＝10577.73-5177.13*25.12/100＝9277.24；
1.3.5.3 2 deuterated (upper 2 deuterated) base peak m/z response value A 2 Response = 292.2
-A 0 *C/100-A 1 *B/100＝1953723.62-5177.13*2.82/100-9277.24*25.12/100＝1951247.18；
1.3.5.4 3 deuterated (upper 3 deuterated) base peak m/z response value A 3 Response = 293.2
-A 1 *C/100-A 2 *B/100＝492863.69-9277.24*2.82/100-1951247.18*25.12/100＝2448.78；
1.3.5.5 4 deuterated (upper 4 deuterated) base peak m/z response value A 4 Response = 294.2
-A 2 *C/100-A 3 *B/100＝55068.71-1951247.18*2.82/100-2448.78*25.12/100＝-11.594；
1.3.5.6 5 deuterated (upper 5 deuterated) base peak m/z response value A 5 Response =295.2
-A 3 *C/100-A 4 *B/100＝98.26-2448.78*2.82/100＝29.20；
1.3.5.6 results to obtain A 0 ＝5177.13；A 1 ＝9277.24；A 2 ＝1951247.18；A 3 ＝2448.78；A 4 ＝-11.594；
A 5 ＝29.20。
1.3.5.7 results discussion: a is that 4 The calculated values appear negative, demonstrating that there is no 4 deuteration and the GC-MS results appear to be suspected of 4 deuteration base peak 294.2 being 2 deuteration and 3 deuteration isotope responses; a is that 5 The abundance percentage is only 0.01%, and it can be judged that 295.2, which is suspected of being the 5 deuterated base peak, is the 3 deuterated isotope response.
1.3.5.8 the individual non-negative base peak response values are then summed, i.e
K＝A 0 +A 1 +A 2 +A 3 ＝5177.13+9277.24+1951247.18+2448.78＝1968150.33。
1.3.5.9 the ratio of each deuteration in sample 1 after deuteration:
1.3.5.10 non-deuterated=a 0 /K*100％＝5177.13/1968150.33*100％＝0.26％；
1.3.5.11 1 deuteration (1 deuterium above) =a 0 /K*100％＝9277.24/1968150.33*100％＝0.47％；
1.3.5.12 2 deuteration (upper 2 deuterium) =a 0 /K*100％＝1951247.18/1968150.33*100％＝99.14％；
1.3.5.13 3 deuteration (3 deuterium above) =a 0 /K*100％＝2448.78/1968150.33*100％＝0.12％；
Namely, the ratio of the target compound in the sample 1 after deuteration is not subjected to deuterium: 0.26%, the upper 1 deuterium ratio of 0.47%, the upper 2 deuterium ratio of 99.14%, and the upper 3 deuterium ratio of 0.12%.
1.3.6 from the results of fig. 3, the corresponding values of the deuterated base peaks possibly present in deuterated sample 2 were calculated according to the following formula:
1.3.6.10 deuterated group peak m/z response value A 0 ＝192091.78；
1.3.6.2 1 deuterated (upper 1 deuterium) base peak m/z response value A 1 Response value=291.2
-A 0 *B/100＝65962.43-192091.78*25.12/100＝17708.97；
1.3.6.3 2 deuterated (upper 2 deuterated) base peak m/z response value A 2 Response = 292.2
-A 0 *C/100-A 1 *B/100＝1600078.88-192091.78*2.82/100-17708.97*25.12/100＝1590213.40；
1.3.6.4 3 deuterated (upper 3 deuterated) base peak m/z response value A 3 Response = 293.2
-A 1 *C/100-A 2 *B/100＝399935.88-17708.97*2.82/100-1590213.40*25.12/100＝-25.12；
1.3.6.5 4 deuterated (upper 4 deuterated) base peak m/z response value A 4 Response = 294.2
-A 2 *C/100-A 3 *B/100＝44794.23-2448.78*25.12/100＝-43.48；
1.3.6.6 result to obtain A 0 ＝192091.78；A 1 ＝17708.97；A 2 ＝1590213.40；A 3 ＝-25.12；A 4 ＝-43.48。
1.3.6.7 discussion: a is that 3 The calculated values appear negative, demonstrating that there is no 3 deuteration (3 deuterations above), and the GC-MS results appear to be suspected 3 deuteration base peak 293.2 as 1 deuteration and 2 deuteration isotope responses; a is that 4 The calculated values appear negative, demonstrating that there is no 4 deuteration (4 deuterations above), and the GC-MS result is suspected of having a 4 deuteration base peak 294.2 as a 2 deuterated isotopic response.
1.3.6.8 the individual non-negative base peak response values are then summed, i.e. k=a 0 +A 1 +A 2
＝192091.78+17708.97+1590213.40＝1800014.15。
1.3.6.9 the ratio of deuterated groups of sample 1 after deuteration:
1.3.6.10 non-deuterated = a 0 /K*100％＝192091.78/1800014.15*100％＝10.67％；
1.3.6.11 1 deuteration=a 0 /K*100％＝17708.97/1800014.15*100％＝0.89％；
1.3.6.12 2 deuteration=a 0 /K*100％＝1590213.40/1800014.15*100％＝88.34％；
I.e., the ratio of non-deuterated in deuterated sample 2: 10.67%, the upper 1 deuterium ratio is 0.89%, and the upper 2 deuterium ratio is 88.34%.
1.4 methodological verification
1.4.1 instrument stability verification
The deuterated pre-sample prepared in step 1.2 was continuously fed into 3 needles using the analytical method of 1.3.1:
table 1: instrument stability verification result summarization
As a result, three continuous needles are fed to the same bottle of sample solution, the RSD of the main peak base peak response value and the main peak isotope peak abundance percentage is less than 1%, and the instrument stability is good.
1.4.2 solution stability verification
The deuterated pre-sample prepared in step 1.2 was placed in a laboratory environment (temperature: 15-30 ℃ C., humidity 20% -80%) and tested by the analytical method of 1.3.1 at 0,2, 4, 8 hours, and a total of 8 hours solution stability test was performed, the results are shown in Table 2 below:
table 2: summarizing the stability verification results of the sample solution
As a result, the sample solution of the same bottle is placed in a laboratory environment (the temperature is 15-30 ℃ and the humidity is 20-80%), the RSD of the main peak base peak response value and the main peak isotope peak abundance percentage is less than 1% in 8 hours, and the sample solution is proved to have good stability in 8 hours in the laboratory environment.
1.4.3 method stability verification
The same deuterated samples were analyzed on the first, second and third days, respectively, using chromatographic conditions of 1.3.1, in triplicate every day, and the measured deuteration rate results are shown in table 3:
table 3: method stability verification result summarization
As a result, the relative standard deviation of the deuteration rate results measured in three days is less than 2.0%, and the method has good reproducibility.
1.4.4 method accuracy verification
1.4.4.1 the HNMR is used to confirm deuterated samples, and it is known from HNMR diagram that two of the cyclopentyl groups connected to benzene rings in the compound are replaced by deuterium, see fig. 4, and the other positions of H are unchanged, i.e. the compound is a 2 deuterated (upper 2 deuterated) compound, see fig. 5.
1.4.4.2 the 2 deuterated (upper 2 deuterium) samples described above were tested using chromatographic conditions of 1.3.1 and from the total ion flow it can be seen that the samples were substantially free of other impurities, see fig. 6.
1.4.4.3 to the non-deuterated sample, the above 2 deuterated (upper 2 deuterated) samples were added in amounts shown in the following table, each group was added in parallel, the ratio of 2 deuteration was calculated by using a formula by detecting the chromatographic condition of 1.3.1, and the recovery rate was calculated, and the results are shown in the following table.
Table 4: summarizing results of method accuracy verification
Results: the recovery rate of the 2 deuterated (upper 2 deuterated) samples added into the non-deuterated samples from low to high is proved to be 94.35-99.56%, the recovery rate is good, and the method accuracy is high.
In conclusion, the method for detecting the deuteration rate of the substance by using the GC-MS has low requirements on detection instruments, can evaluate the specific deuterium number, and has important guiding significance on production.
Claims (7)
1. A method for assaying deuteration of a substance by GC-MS comprising the steps of:
1) Weighing a certain amount of pre-deuterated sample and a certain amount of post-deuterated sample to be detected, adding a proper amount of solvent to completely dissolve the sample, and preparing a sample blank;
2) Taking a proper amount of the solution in the step 1), passing through an organic filter membrane, and bottling and measuring the solution as an upper sample solution;
3) Detecting the sample solution in the step 2) by using GC-MS;
4) Integrating main peaks of the samples before and after deuteration after deducting blank according to the result obtained by GC-MS detection in the step 3), and extracting ions, response values and abundance percentages of the main peaks;
5) Extracting main peak ions before and after deuteration, response values and abundance percentage data in the step 4), and calculating the deuteration rate of the sample;
the structure of the deuterated pre-sample is shown as formula I, and the molecular formula is C 22 H 26 The theoretical m/z is 290.20,291.21,292.21 respectively; the abundance percentage is 100.0%,24.1% and 2.8% respectively;
the ion group peak result of the main peak molecule before deuteration in the step 4) is marked as M, and the isotopic abundance percentage result of the main peak molecule before deuteration is marked as 100:B:C;
the method for calculating the deuteration rate of the sample comprises the following 4 steps:
(1) According to the molecular ion group peak M of the sample before deuteration, the molecular ion group peak M/z of non-deuteration is obtained as follows: m+0; the ionic group peak m/z of 1 deuterated (upper 1 deuterium) molecule is: m+1; the 2 deuterated (upper 2 deuterated) molecular ionic groups peak m/z is: m+2;3 deuterated (upper 3 deuterated) molecular ionic groups peak m/z is: m+3; the ion group peak m/z of 4 deuterated (4 deuterium) molecules is: m+4; the 5 deuterated (upper 5 deuterated) molecular ion groups peak m/z is: m+5; … …;
(2) And calculating the response value of each deuterated base peak possibly existing in the deuterated sample, wherein the calculation formula is as follows: 0 deuterated group peak M has response value A 0 The method comprises the steps of carrying out a first treatment on the surface of the 1 deuterated group peak m/z response valueA 1 Response value-a=m+1 0 * B/100;2 deuterated group peak m/z response value A 2 Response-a =m+2 0 *C/100-A 1 * B/100;3 deuterated group peak m/z response value A 3 Response-a =m+3 1 *C/100-A 2 * B/100;4 deuterated group peak m/z response value A 4 Response-a =m+4 2 *C/100-A 3 * B/100;5 deuterated group peak m/z response value A 5 Response =m+5
-A 3 *C/100-A 4 * B/100 … …; until a negative value occurs in the result;
(3) The deuterated non-negative base peak response values are then summed, i.e., A 0 +A 1 +A 2 +A 3 +A 4 +A 5 +……＝K；
(4) Calculating the ratio of each deuterated to the deuterated sample: non-deuteration=a 0 K is 100%,1 deuteration=a 1 K is 100%,2 deuteration=a 2 K is 100%,3 deuteration=a 3 K is 100%,4 deuteration=a 4 K is 100%,5 deuteration=a 5 /K*100％……。
2. The method for detecting the deuteration rate of a substance by using GC-MS according to claim 1, wherein the method in the step 1) is to respectively weigh a sample before deuteration and about 10mg of deuteration to be detected, and add a solvent with corresponding volume to dissolve the sample and the deuteration to prepare 1mg/mL of the upper solution; the sample before deuteration is a sample before deuteration of the product; the blank sample is a blank solution prepared by the same steps as the sample except that no sample is added.
3. The method for determining deuteration rate of material according to claim 2 wherein said solvent is dichloromethane or tetrahydrofuran, said solution is bottled in 20mL brown glass bottles and said dissolving is shaking or sonicating until the sample is completely dissolved.
4. A method for determining the deuteration rate of a material according to claim 3 wherein said solvent is methylene chloride.
5. The method for detecting deuteration rate of material according to claim 4 wherein said organic filter is a 0.45 μm nylon needle filter and wherein said filtrate is contained in a 2mL brown sample bottle for chromatography.
6. The method for detecting deuteration rate of a material according to claim 5 wherein said step 3) comprises the steps of: gasification chamber temperature: 320 ℃; column flow rate: 1.2ml/min; column temperature: maintaining at 100deg.C for 1min, heating to 250deg.C at 10deg.C/min, and maintaining for 1min; heating to 320 ℃ at 20 ℃/min and keeping for 9.5min; split ratio 30:1, a step of; chromatographic column: agilent HP-5ms UI 30m x 250 μm x 0.25 μm; MS interface temperature: 330 ℃; intuvo flow path: 320 ℃; the sample injection amount is as follows: 1 microliter; ion source: EI; electron energy: 70eV, MS ion source 230 ℃; MS quadrupole rod 150 ℃; solvent delay time is 3min, the solvent A dichloromethane and the solvent B methanol are respectively used for 5 times before and after sample injection, the sample to be injected is used for 3 times before sample injection, then suction is carried out for 4 times, the bubbles of the sample injection needle are removed, and carrier gas is generated: high purity helium, ion scan range: 35-350.
7. The method for determining deuteration of a material by GC-MS according to claim 1 wherein the main peak retention time in step 4) is 19.5 ± 0.5min.
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