CN114324720A - Method for detecting isomer in levo-hydrochloric acid demethyl phencynonate - Google Patents
Method for detecting isomer in levo-hydrochloric acid demethyl phencynonate Download PDFInfo
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- -1 demethyl phencynonate Chemical compound 0.000 title claims abstract description 19
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Abstract
The invention provides a method for detecting isomers in levo-hydrochloric acid demethyl phencynonate, which comprises the steps of preparing a blank solution, preparing a sample solution, preparing a test solution, detecting by a high performance liquid chromatography method and the like, and can qualitatively or quantitatively detect the content of dextro-hydrochloric acid demethyl phencynonate in levo-hydrochloric acid demethyl phencynonate. The method has the characteristics of strong specificity, high accuracy, good reproducibility and accurate and reliable detection result, can control the quality of the levorotatory hydrochloric acid demethylated phencynonate, and improves the medication safety of patients.
Description
Technical Field
The invention belongs to the field of drug analysis, and particularly relates to a method for detecting an isomer in levorotatory hydrochloric acid demethylated phencynonate.
Background
The levo-phencynonate hydrochloride is a metabolite of levo-phencynonate, is an M4 receptor selective antagonist with a brand-new structure, and has selectivity indexes (M4/M1 is more than 100, M4/M2 is more than 100, M4/M3 is more than 10, and M4/M5 is more than 10) superior to M4 receptor selective antagonists reported in the prior literature. Because of the characteristics of strong activity (IC50 value is 4.4nmol/L) and low toxicity (LD 50 value is 490mg/kg for mice), the compound has been used as a candidate drug for resisting Parkinson disease and enters preclinical research.
Dextrorotatory phencynonate may be generated in the synthesis process of levorotatory phencynonate hydrochloride, so as to provide a safer and more effective novel medicine for treating parkinsonism for clinic, promote the development of M receptor antagonist and chiral medicine in PD treatment application, and ensure the medication safety of patients, the isomer dextrorotatory phencynonate hydrochloride in the levorotatory phencynonate hydrochloride needs to be detected. However, no method for measuring isomer of demethylated cyclononyl levlate hydrochloride in the same has been reported in domestic and foreign literature. Therefore, it is necessary and urgent to establish an analytical method capable of determining isomer of demethylated cyclononyl levulinate hydrochloride in demethylated cyclononyl levulinate hydrochloride.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide the high performance liquid chromatography which has strong specificity, high accuracy, good reproducibility and accurate and reliable detection result and can detect the isomer of the demethylated cyclononyl levulinate hydrochloride in the demethylated cyclononyl levulinate hydrochloride, thereby controlling the quality of the demethylated cyclononyl levulinate hydrochloride and improving the medication safety of patients.
The application is realized by the following technical scheme:
the application provides a method for detecting an isomer in levorotatory demethylated phencynonate hydrochloride, wherein the isomer is dextrorotatory demethylated phencynonate hydrochloride, and the method comprises the following steps:
(1) preparing a blank solution: taking acetonitrile and pure water, and fully and uniformly mixing to obtain a blank solution;
(2) preparing a sample solution: taking a levo-hydrochloric acid demethylated phencynonate sample, adding a diluent to completely dissolve the levo-hydrochloric acid demethylated phencynonate sample to obtain a sample solution;
(3) and (3) measuring the content: respectively measuring blank solution and sample solution, injecting into a high performance liquid chromatograph, recording chromatogram, and calculating the content of dextrorotatory phencynonate hydrochloride isomer according to an area normalization method.
Further, the chromatographic conditions of the high performance liquid chromatograph are as follows:
a chromatographic column: agilent Technologies HPLC-cassette 250-4ChiraDex, 5 μm;
the detection wavelength is as follows: 220 nm;
the sample injection volume is as follows: 5-50 mu L;
the column box temperature was: 20-35 ℃;
the flow rate is: 0.6mL/min-1.0 mL/min;
the mobile phase comprises an A phase and a B phase, the volume ratio of the A phase to the B phase is 30: 70-40: 60,
the phase A is a mixed solution of acetonitrile and methanol, and the phase B is a potassium dihydrogen phosphate solution with the mass molar concentration of 0.01 mol/L.
Further, the flow rates are: 0.8 mL/min.
Further, the column box temperature is: at 25 ℃.
Further, the volume ratio of the phase a to the phase B was 35:65, and the volume ratio of methanol to acetonitrile in the phase a was 25: 10.
Further, the preparation method of the dextro-rotatory hydrochloric acid demethyl phencynonate stock solution comprises the following steps:
taking about 10mg of dextro-rotatory norbencyclononyl hydrochloride reference substance, precisely weighing, placing into a 100ml measuring flask, adding about 2/3 volume of diluent to completely dissolve, diluting to constant volume with the diluent, and shaking up; precisely measuring 10ml, putting into a 250ml measuring flask, diluting to constant volume with diluent, and shaking up to obtain the dextrorotatory norbencyclononyl hydrochloride stock solution with the concentration of 4.0 mug/ml.
Further, before the content is measured in the step (3), a system applicability experiment is also included, and the method specifically comprises the following steps: fully mixing the left-handed hydrochloric acid demethylated phencynonate stock solution and the right-handed hydrochloric acid demethylated phencynonate stock solution to obtain a system applicability solution; measuring the system applicability solution, injecting into a high performance liquid chromatograph, and measuring the separation degree of the peaks of the levorotatory demethylated phencynonate hydrochloride and the dextrorotatory demethylated phencynonate hydrochloride.
Furthermore, the separation degree of the peaks of the levorotatory demethylated phencynonate hydrochloride and the dextrorotatory demethylated phencynonate hydrochloride is more than 1.6.
Further, the preparation method of the L-hydrochloric acid demethylated phencynonate stock solution comprises the following steps: and (2) taking 80mg of a levorotatory norbencyclononyl hydrochloride reference substance, precisely weighing, placing into a 100ml measuring flask, adding a diluent to completely dissolve, adding the diluent to a constant volume, and shaking up to obtain the levorotatory norbencyclononyl hydrochloride stock solution, wherein the concentration of the levorotatory norbencyclononyl hydrochloride stock solution is 800 mu g/ml.
Further, the diluent is 60% acetonitrile.
The beneficial effect of this application lies in: the method has the advantages of strong specificity, high accuracy, good reproducibility and accurate and reliable detection result, can qualitatively or quantitatively detect the content of dextrorotatory phencynonate in levorotatory phencynonate hydrochloride, is convenient for controlling the quality of the levorotatory phencynonate hydrochloride, and provides practical and effective guarantee for improving the medication safety of patients.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required to be used in the embodiments will be briefly described below.
FIG. 1 is a high performance liquid chromatogram of an air-white solution in example 3.
FIG. 2 is a high performance liquid chromatogram of the resolution solution of example 4.
FIG. 3 is a high performance liquid chromatogram of the L-S-1 solution in example 6.
FIG. 4 is a high performance liquid chromatogram of a solution A-S-1-1 of the test sample (0.2% demethylated phencynonate D-HCl) in example 7.
FIG. 5 is a high performance liquid chromatogram of a solution A-S-2-1 of the sample (0.5% demethylated phencynonate D-HCl) in example 7.
FIG. 6 is a high performance liquid chromatogram of a solution A-S-3-1 of the sample (0.6% demethylated phencynonate D-HCl) in example 7.
Detailed Description
The present invention is described in further detail below with reference to specific examples, which are not to be construed as limiting the scope of the invention as claimed herein.
Example 1:
the method for detecting the isomer in the levo-hydrochloric acid demethyl phencynonate comprises the following steps:
1. solution preparation:
blank solution: taking 600ml of acetonitrile and 400ml of purified water, mixing uniformly, and degassing to obtain the product.
Dextro-phencynonate hydrochloride stock solution (4.0 mug/ml): taking about 10mg of dextro-rotatory norbencyclononyl hydrochloride reference substance, precisely weighing, placing into a 100ml measuring flask, adding about 2/3 volume of diluent to completely dissolve, diluting to constant volume with the diluent, and shaking up; precisely measuring 10ml, placing into a 250ml measuring flask, diluting to constant volume with diluent, and shaking up to obtain the final product.
L-Phenylcyclononyl Dehydrochlorid stock solution (800. mu.g/ml): taking about 80mg of a levo-hydrochloric acid demethyl phencynonate reference substance, precisely weighing, placing into a 100ml measuring flask, adding about 2/3 volume of diluent for complete dissolution, adding the diluent for constant volume, and shaking up to obtain the levo-hydrochloric acid demethyl phencynonate contrast substance.
Resolution solution: precisely measuring 2.5ml of dextrorotatory phencynonate hydrochloride stock solution and 2.5ml of levorotatory phencynonate hydrochloride stock solution into a 5ml volumetric flask, and shaking uniformly.
Sample solution: taking a levo-hydrochloric acid demethylated phencynonate sample of 20mg, precisely weighing, placing in a 50ml volumetric flask, dissolving by using a diluent, fixing the volume, and shaking up;
the diluent in the solution formulation is 60% acetonitrile.
2. And (3) determination: and respectively carrying out high performance liquid chromatograph detection on the sample solution and the blank solution, and recording chromatograms. The chromatographic conditions of the high performance liquid chromatograph are as follows:
a chromatographic column: agilent Technologies HPLC-cassette 250-4ChiraDex, 5 μm chiral chromatography column;
the detection wavelength is as follows: 220 nm;
the sample injection volume is as follows: 20 mu L of the solution;
the column box temperature was: 25 ℃;
the flow rate is: 0.8 mL/min;
the mobile phase comprises an A phase and a B phase, the volume ratio of the A phase to the B phase is 30: 70-40: 60,
the phase A is a mixed solution of acetonitrile and methanol, the phase B is a potassium dihydrogen phosphate solution with the mass molar concentration of 0.01mol/L, the volume ratio of the phase A to the phase B is 35:65, and the volume ratio of the methanol to the acetonitrile in the phase A is 25: 10.
Example 2 gas chromatography conditions are shown in table 1:
TABLE 1
| Chromatographic column | Agilent Technologies HPLC-Cartridge 250-4ChiraDex(5μm) |
| Detection wavelength | 220nm |
| Detector | UV |
| Flow rate of flow | 0.8ml/min |
| Sample introduction volume | 20μl |
| |
25℃ |
| Sample pan temperature | At room temperature |
| Run time | 30min |
| Mobile phase | Methanol-acetonitrile-0.01 mol/L potassium dihydrogen phosphate solution (phosphoric acid, pH3.2) (25:10:65) |
Example 3 specificity experiments
Blank solution: taking 600ml of acetonitrile and 400ml of purified water, mixing uniformly, and degassing to obtain the product. And (5) detecting the blank solution by using a high performance liquid chromatograph, and recording a chromatogram. The high performance liquid chromatogram of the blank solution is shown in FIG. 1.
See example 2 for high performance liquid chromatography conditions.
And (4) conclusion: as can be seen from FIG. 1, the blank solution does not interfere with the sample to be tested.
Example 4 degree of separation experiment
Dextro-phencynonate hydrochloride stock solution (4.0 mug/ml): taking about 10mg of dextro-rotatory norbencyclononyl hydrochloride reference substance, precisely weighing, placing into a 100ml measuring flask, adding about 2/3 volume of diluent to completely dissolve, diluting to constant volume with the diluent, and shaking up; precisely measuring 10ml, placing into a 250ml measuring flask, diluting to constant volume with diluent, and shaking up to obtain the final product.
L-Phenylcyclononyl Dehydrochlorid stock solution (800. mu.g/ml): taking about 80mg of a levo-hydrochloric acid demethyl phencynonate reference substance, precisely weighing, placing into a 100ml measuring flask, adding about 2/3 volume of diluent for complete dissolution, adding the diluent for constant volume, and shaking up to obtain the levo-hydrochloric acid demethyl phencynonate contrast substance.
Resolution solution: precisely measuring 2.5ml of dextrorotatory phencynonate hydrochloride stock solution and 2.5ml of levorotatory phencynonate hydrochloride stock solution into a 5ml volumetric flask, and shaking uniformly.
And detecting the resolution solution by using a high performance liquid chromatograph, and recording a chromatogram. The high performance liquid chromatogram of the resolution solution is shown in FIG. 2.
See example 2 for high performance liquid chromatography conditions.
And (4) conclusion: as can be seen from FIG. 2, the separation degree of the peaks of the L-demethyl phencynonate hydrochloride and the D-demethyl phencynonate hydrochloride is 1.925. When the separation degree is 1.6 in the high performance liquid chromatography analysis, the separation degree is considered to reach the baseline separation, and the standard separation degree which can obtain the most accurate quantitative analysis result can be ensured, the separation degree of the peaks of the levorotatory demethylated phencynonate hydrochloride and dextrorotatory demethylated phencynonate hydrochloride is 1.925 which is far more than 1.6, and therefore, the stable analysis can be ensured.
Example 5 Linear experiment
Solutions were prepared at the following concentrations, respectively:
L-S-1 (0.2%, dextro-phencynonate hydrochloride): precisely measuring 2ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, performing constant volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution.
L-S-2 (0.3%, dextro-phencynonate hydrochloride): precisely measuring 3ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, performing constant volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution.
L-S-3 (0.4%, dextro-phencynonate hydrochloride): precisely measuring 4ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, fixing the volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution.
L-S-4 (0.5%, dextro-phencynonate hydrochloride): precisely measuring 5ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, fixing the volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution.
L-S-5 (0.6%, dextro-phencynonate hydrochloride): precisely measuring 6ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, fixing the volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution.
L-R-1 (40%, demethylphencynonate levo-hydrochloride): precisely measuring 2ml of the L-demethyl phencynonate hydrochloride stock solution, putting the L-demethyl phencynonate stock solution into a 10ml measuring flask, diluting to a constant volume by using a diluent, and uniformly mixing to obtain the L-demethyl phencynonate hydrochloride stock solution.
L-R-2 (60%, demethylphencynonate levo-hydrochloride): precisely measuring 3ml of the L-demethyl phencynonate hydrochloride stock solution, putting the L-demethyl phencynonate stock solution into a 10ml measuring flask, diluting to a constant volume by using a diluent, and uniformly mixing to obtain the L-demethyl phencynonate hydrochloride stock solution.
L-R-3 (80%, demethylphencynonate L-HCl): precisely measuring 4ml of the L-decylnyloxinyl hydrochloride stock solution, putting the L-decylnyloxinyl hydrochloride stock solution into a 10ml measuring flask, diluting to a constant volume with a diluent, and uniformly mixing to obtain the final product.
L-R-4 (100%, demethylphencynonate L-HCl): precisely measuring 5ml of the L-norphencynonate hydrochloride stock solution, putting the L-norphencynonate hydrochloride stock solution into a 10ml measuring flask, diluting to a constant volume by using a diluent, and uniformly mixing to obtain the L-norphencynonate hydrochloride.
L-R-5 (120%, demethylphencynonate L-HCl): precisely measuring 6ml of the levorotatory demethylated phencynonate hydrochloride stock solution, putting the levorotatory demethylated phencynonate stock solution into a 10ml measuring flask, diluting to a constant volume by using a diluent, and uniformly mixing to obtain the levorotatory demethylated phencynonate hydrochloride stock solution.
See example 2 for high performance liquid chromatography conditions.
The results of the linear experimental determination of dextro-rotatory demethyl phencynonate hydrochloride are shown in the table 2:
the linear equation is: 17711.5010x-862.8000
Correlation coefficient R is 0.9982
Intercept percentage: 0.0122%
TABLE 2
The results of the linear experimental determination of the levorotatory hydrochloric acid demethyl phencynonate are shown in the table 3:
the linear equation is: 17907.9517x +3603.2000
Correlation coefficient R:1.0000
Intercept percentage: 0.0503 percent
TABLE 3
And (4) conclusion: the linear experimental determination result of the dextrorotatory demethylated phencynonate hydrochloride and the linear experimental determination result of the levorotatory demethylated phencynonate hydrochloride can both be known, and the concentration of each solvent and the peak area form a good linear relation, thereby meeting the linear determination requirement.
Example 6 quantitative Limit experiment
Solution preparation: the same procedure as for the L-S-1 solution in example 5 (0.2%, dextrorotatory demethyl phencynonate hydrochloride): precisely measuring 2ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, performing constant volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution. See example 2 for high performance liquid chromatography conditions. The high performance liquid chromatogram of the L-S-1 solution is shown in FIG. 3.
As a result: as can be seen from the high performance liquid chromatogram of fig. 3, the concentrations are: 0.8 mu g/ml; 20.51 of S/N; 1.03 percent of RSD;
and (4) conclusion: the quantitative limit meets the requirement, and the dextro-rotatory hydrochloric acid demethyl phencynonate can be accurately quantified about the limit.
Example 7 accuracy experiment
Solution preparation
Accuracy experiments were performed with a blank solution as background.
Test solution (0.2%, dextro-rotatory phencynonate hydrochloride): precisely measuring 2ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, performing constant volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution. In parallel 3 parts. Wherein: the high performance liquid chromatogram of the test solution (0.2%, dextro-rotatory phencynonate hydrochloride) A-S-1-1 solution is shown in FIG. 4.
Test solution (0.5%, dextro-rotatory phencynonate hydrochloride): precisely measuring 5ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, fixing the volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution. In parallel 3 parts. Wherein: the high performance liquid chromatogram of the test solution (0.5%, dextro-rotatory phencynonate hydrochloride) A-S-2-1 solution is shown in FIG. 5.
Test solution (0.6%, dextro-rotatory phencynonate hydrochloride): precisely measuring 6ml of dextrorotatory phencynonate hydrochloride stock solution, placing the dextrorotatory phencynonate hydrochloride stock solution into a 10ml measuring flask, fixing the volume by using a diluent, and uniformly mixing to obtain the dextrorotatory phencynonate hydrochloride stock solution. In parallel 3 parts. Wherein: the high performance liquid chromatogram of the test solution (0.6% of dextro-phencynonate hydrochloride) A-S-3-1 is shown in FIG. 6.
The HPLC conditions are shown in example 2, and the results of the accuracy test are shown in Table 4.
And (4) conclusion: the average recovery rates of the test solution (0.2 percent, dextrorotation demethylated phencynonate hydrochloride), the test solution (0.5 percent, dextrorotation demethylated phencynonate hydrochloride) and the test solution (0.6 percent, dextrorotation demethylated phencynonate hydrochloride) are respectively 105.4 percent, 103.9 percent and 105.2 percent, and the RSD recovery rates are respectively 2.15 percent, 0.61 percent and 1.17 percent, which indicates that the method has better accuracy.
TABLE 4
In conclusion, the method for detecting the isomer in the L-hydrochloric acid demethyl phencynonate is determined by the preparation conditions of the sample solution, the preparation conditions of the test solution and the chromatographic conditions. The detection method for the isomer in the levo-hydrochloric acid demethylated cyclononyl ester has the advantages of strong specificity, high accuracy, good reproducibility and accurate and reliable detection result, can qualitatively or quantitatively detect the content of dextro-hydrochloric acid demethylated cyclononyl ester in the levo-hydrochloric acid demethylated cyclononyl ester, is convenient to control the quality of the levo-hydrochloric acid demethylated cyclononyl ester, and provides practical and effective guarantee for improving the medication safety of patients.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. A method for detecting an isomer in demethylated phencynonate levorotatory hydrochloride is characterized in that the isomer is dextrorotatory phencynonate dextrorotatory hydrochloride, and the method comprises the following steps:
(1) preparing a blank solution: taking acetonitrile and pure water, and fully and uniformly mixing to obtain a blank solution;
(2) preparing a sample solution: taking a levo-hydrochloric acid demethylated phencynonate sample, adding a diluent to completely dissolve the levo-hydrochloric acid demethylated phencynonate sample to obtain a sample solution;
(3) and (3) measuring the content: respectively measuring blank solution and sample solution, injecting into a high performance liquid chromatograph, recording chromatogram, and calculating the content of dextrorotatory phencynonate hydrochloride isomer according to an area normalization method.
2. The method for detecting isomers of demethylated cyclononyl levorotatory hydrochloride according to claim 1, wherein the chromatographic conditions of said HPLC are as follows:
a chromatographic column: agilent Technologies HPLC-cassette 250-4ChiraDex, 5 μm;
the detection wavelength is as follows: 220 nm;
the sample injection volume is as follows: 5-50 mu L;
the column box temperature was: 20-35 ℃;
the flow rate is: 0.6mL/min-1.0 mL/min;
the mobile phase comprises an A phase and a B phase, the volume ratio of the A phase to the B phase is 30: 70-40: 60,
the phase A is a mixed solution of acetonitrile and methanol, and the phase B is a potassium dihydrogen phosphate solution with the molar concentration of 0.01 mol/L.
3. The method for detecting isomers of norbencyclononyl levohcitrate according to claim 2, wherein the flow rates are as follows: 0.8 mL/min.
4. The method for detecting isomers of norbencyclononyl levo-hydrochloride according to claim 2, wherein the column box temperature is: at 25 ℃.
5. The method for detecting isomers of norphencynonate levorotatory hydrochloride according to claim 2, wherein the volume ratio of the phase A to the phase B is 35:65, and the volume ratio of methanol to acetonitrile in the phase A is 25: 10.
6. The method for detecting isomers in l-desmethyl phencynonate hydrochloride according to claim 1, wherein the method for preparing the l-desmethyl phencynonate hydrochloride stock solution is as follows:
taking about 10mg of dextro-rotatory norbencyclononyl hydrochloride reference substance, precisely weighing, placing into a 100ml measuring flask, adding 2/3 volume of diluent to completely dissolve, diluting to constant volume with the diluent, and shaking up; precisely measuring 10ml, putting into a 250ml measuring flask, diluting to constant volume with diluent, and shaking up to obtain the dextrorotatory norbencyclononyl hydrochloride stock solution with the concentration of 4.0 mug/ml.
7. The method for detecting isomers of l-decylncyclononyl levhyhci according to claim 1, wherein before the content is measured in step (3), a systematic applicability test is further included, specifically: fully mixing the left-handed hydrochloric acid demethylated phencynonate stock solution and the right-handed hydrochloric acid demethylated phencynonate stock solution to obtain a system applicability solution; measuring the system applicability solution, injecting into a high performance liquid chromatograph, and measuring the separation degree of the peaks of the levorotatory demethylated phencynonate hydrochloride and the dextrorotatory demethylated phencynonate hydrochloride.
8. The method of claim 7, wherein the peak separation between the desmethylphenylcyclononyl levehydrochloride and the desmethylphenylcyclononyl dextrohcl is greater than 1.6.
9. The method for detecting isomers of demethylated cyclononyl levorotatory hydrochloride according to claim 7, wherein the stock solution of demethylated cyclononyl levorotatory hydrochloride is prepared by the following steps: and (2) taking 80mg of a levorotatory norbencyclononyl hydrochloride reference substance, precisely weighing, placing into a 100ml measuring flask, adding a diluent to completely dissolve, adding the diluent to a constant volume, and shaking up to obtain the levorotatory norbencyclononyl hydrochloride stock solution, wherein the concentration of the levorotatory norbencyclononyl hydrochloride stock solution is 800 mu g/ml.
10. The method for detecting isomers of norbencyclononyl levorotatory hydrochloride according to claim 1, 6 or 9, wherein the diluent is 60% acetonitrile.
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