CN117347527A - Detection method of N-methyl-L-prolyl related substances - Google Patents
Detection method of N-methyl-L-prolyl related substances Download PDFInfo
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- 239000000126 substance Substances 0.000 title claims abstract description 34
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- 238000000034 method Methods 0.000 claims abstract description 23
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- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 6
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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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- 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
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N30/54—Temperature
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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
- G01N30/62—Detectors specially adapted therefor
- G01N30/64—Electrical detectors
- G01N30/68—Flame ionisation detectors
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Abstract
The invention discloses a detection method of N-methyl-L-prolyl related substances, which adopts gas chromatography to measure the related substances, and the chromatographic conditions are as follows: chromatographic column: capillary column with cyanopropyl phenyl-dimethyl polysiloxane as stationary phase; carrier gas flow rate: 0.5 to 1.5ml/min; split ratio: 48-52:1; heating program: the initial temperature is 40-50 ℃, the temperature is maintained for 1-3 min, the temperature is raised to 275-285 ℃ at the rate of 18-22 ℃ per minute, and the temperature is maintained for 2-8 min; sample inlet temperature: 275-285 ℃; the detector is a hydrogen flame ionization detector, and the temperature of the detector is 275-285 ℃. The detection method provided by the invention can realize the effective separation of N-methyl-L-prolyl alcohol, L-prolyl alcohol and N-methyl-L-proline. The method provided by the invention is sensitive, accurate and good in reproducibility, provides reliable guarantee for improving and better controlling the quality of the N-methyl-L-prolyl, and has very important significance for improving the medication safety.
Description
Technical Field
The invention relates to the technical field of medicine analysis and detection, in particular to a detection method of N-methyl-L-prolyl related substances.
Background
N-Methyl-L-prolyl alcohol (English name N-Methyl-L-prolinol, molecular formula: C) 6 H 13 NO, molecular weight: 115.17 Is an agent for the synthesis of novel 4-hydroxy tamoxifen analogues, useful as estrogen-related gamma (errgamma) inverse agonists.
N-methyl-L-prolyl alcohol is an important organic compound and has wide application and research value in the anti-inflammatory field. Anti-inflammatory therapy is one of the important means of treating inflammatory diseases, and N-methyl-L-prolyl has received a great deal of attention as a compound with potential anti-inflammatory activity. Studies have shown that N-methyl-L-prolyl is able to inhibit the occurrence and progression of inflammatory reactions, thereby alleviating symptoms and damage of related diseases.
N-methyl-L-prolinol is of great interest not only in the use in neuroinflammation, but also in pulmonary inflammation. First, neuroinflammation is a common feature of various neurological diseases, such as parkinson's disease, alzheimer's disease, and the like. It was found that N-methyl-L-prolyl can reduce the extent of neuroinflammatory reaction by inhibiting the production of inflammatory factors, thereby improving the symptoms and progression of neurological diseases. Pulmonary inflammation is a common inflammatory disease of the respiratory system, such as asthma, chronic obstructive pulmonary disease, and the like. It was found that N-methyl-L-prolol can reduce inflammatory responses in lung tissue, improve lung function, and reduce peak airway flow and positive end-expiratory pressure. This suggests that N-methyl-L-prolyl alcohol may be a potential drug for the treatment of respiratory inflammation.
In recent years, research into N-methyl-L-prolinol has also been directed to other fields. For example, N-methyl-L-prolyl alcohol has been used in the fields of cardiovascular diseases, renal diseases, digestive system diseases, etc. as a preliminary study result. These findings indicate that N-methyl-L-prolyl may be an anti-inflammatory drug with a broad application potential.
As a medicine, besides a certain curative effect, whether to realize accurate and convenient quantitative analysis is one of the technical preconditions for wide popularization. The impurity research is one of key projects in the medicine quality research, and the content of the impurity research is not only a direct index reflecting the purity of medicines, but also has important safety significance.
At present, no detection method for N-methyl-L-prolyl related substances is reported in the prior art. With the improvement of the requirements of China on the medicine quality control work, the establishment of standards and detection methods of related substances becomes an urgent need for improving and better controlling the quality of N-methyl-L-prolyl.
Disclosure of Invention
The invention provides a detection method of N-methyl-L-prolyl related substances aiming at the technical problems. The method can not only realize the effective separation of N-methyl-L-prolyl alcohol from L-prolyl alcohol and N-methyl-L-prolyl acid, but also accurately detect the content of related substances in the N-methyl-L-prolyl alcohol.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
a detection method of N-methyl-L-prolyl related substances comprises the steps of preparing a solution to be detected by taking 100% methanol as a solvent, and measuring the related substances by using a gas chromatography, wherein the related substances comprise L-prolyl and N-methyl-L-proline, and the chromatographic conditions of the gas chromatography are as follows:
chromatographic column: capillary column with cyanopropyl phenyl-dimethyl polysiloxane as stationary phase;
carrier gas flow rate: 0.5-1.5 ml/min;
split ratio: 48-52:1;
heating program: the initial temperature is 40-50 ℃, the temperature is maintained for 1-3 minutes, the temperature is raised to 275-285 ℃ at the rate of 18-22 ℃ per minute, and the temperature is maintained for 2-8 minutes;
sample inlet temperature: 275-285 ℃;
the detector is a hydrogen flame ionization detector, and the temperature of the detector is 275-285 ℃.
Preferably, a method for detecting N-methyl-L-prolinol-related substances comprises the following steps:
preparing a sample solution, and dissolving N-methyl-L-prolyl alcohol by using a solvent to obtain the sample solution;
preparing a system applicability solution, namely dissolving an N-methyl-L-prolyl reference substance and an impurity reference substance stock solution by using a solvent to obtain the system applicability solution; the impurity reference substance stock solution is prepared by dissolving an impurity reference substance with a solvent, wherein the impurities comprise L-prolyl and N-methyl-L-proline;
detection was performed by gas chromatography under the following conditions:
chromatographic column: agilent DB-1701 30m X0.32 mm,0.25 μm;
carrier gas flow rate: 1.0mL/min;
split ratio: 50:1;
heating program: the initial temperature is 45 ℃, maintained for 2 minutes, and the temperature is raised to 280 ℃ at the rate of 20 ℃ per minute, and maintained for 5 minutes;
sample inlet temperature: 280 ℃;
detector temperature: 280 ℃.
Preferably, the method further comprises the following steps: and calculating the content of the N-methyl-L-prolyl related substances in the test sample solution according to the measured data by an external standard method and peak areas.
Preferably, the chromatographic column is a capillary column with 14% cyanopropylphenyl-86% dimethylpolysiloxane as the stationary phase.
Preferably, the limit of L-prolyl alcohol is less than or equal to 0.5 percent.
Preferably, the limit of N-methyl-L-proline is less than or equal to 0.5%.
The beneficial effects of the invention are at least as follows: according to the method, derivatization treatment of N-methyl-L-prolyl alcohol is not needed, a chiral chromatographic column is not needed, a capillary column taking cyanopropyl phenyl-dimethylpolysiloxane as a stationary phase is taken as a chromatographic column, 100% methanol is taken as a solvent to prepare a solution to be detected, and related substances are measured in a direct solution injection mode by controlling a temperature-raising program of a gas chromatography, so that effective separation of the N-methyl-L-prolyl alcohol from the L-prolyl alcohol and the N-methyl-L-proline is realized, and the content of the related substances in the N-methyl-L-prolyl alcohol is accurately detected.
The method provided by the invention is subjected to methodology researches and verification such as specificity, sensitivity and the like, has high sensitivity, better accuracy and reproducibility, can simply, conveniently and quickly realize accurate quantitative detection of N-methyl-L-prolyl related substances, overcomes the defect that the detection of the N-methyl-L-prolyl related substances is not recorded in the prior art, provides reliable guarantee for better controlling the quality of the N-methyl-L-prolyl, and has very important significance for improving the medication safety.
The invention can realize the effective separation of N-methyl-L-prolyl alcohol, L-prolyl alcohol and N-methyl-L-proline without chiral chromatographic columns.
The method of the invention ensures that each component has good peak shape, separation degree and detection sensitivity, has small baseline interference, is more beneficial to effectively separating N-methyl-L-prolyl from L-prolyl and N-methyl-L-proline, and has accurate and reliable result and good repeatability.
The invention effectively improves the separation degree and the detection sensitivity between the N-methyl-L-prolyl alcohol and the N-methyl-L-proline, so that the detection result is accurate in quantification and high in precision.
Drawings
FIG. 1 is a chromatogram of a blank solvent under the proprietary and system applicability terms in the examples;
FIG. 2 is a chromatogram of a system applicability solution under the specificity and system applicability terms in the examples;
FIG. 3 is a chromatogram of a test solution under the specific and system applicability terms in the examples;
FIG. 4 is a chromatogram under detection by gas chromatography in comparative example 1;
FIG. 5 compares the chromatograms under detection by gas chromatography in example 2;
FIG. 6 compares the chromatograms under detection by gas chromatography in example 3;
FIG. 7 is a chromatogram of comparative example 4 under detection by gas-liquid chromatography.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Examples
The embodiment of the invention provides a detection method of N-methyl-L-prolyl related substances
Instrument and reagent
Instrument: gas chromatograph, hydrogen flame ionization detector, measuring flask, electronic balance.
Blank reagent: 100% methanol.
Preparation of the solution
L-prolyl impurity control stock solution: taking a proper amount of L-prolyl reference substance, precisely weighing, and preparing the L-prolyl reference substance into a stock solution containing 16mg of L-prolyl in 1mL of L-prolyl impurity by 100% methanol.
N-methyl-L-proline impurity control stock solution): taking a proper amount of N-methyl-L-proline reference substance, precisely weighing, and preparing an N-methyl-L-proline impurity stock solution with 100% methanol in a concentration of 16mg per 1mL of N-methyl-L-proline impurity stock solution.
System applicability solution: taking a proper amount of N-methyl-L-prolyl reference substance, precisely weighing a proper amount of impurity reference substance stock solution, and preparing a system applicability solution containing 1120 mug of L-prolyl, 1120 mug of N-methyl-L-proline and 224mg of N-methyl-L-prolyl per 1mL of system applicability solution by using 100% methanol.
Test solution: taking a proper amount of N-methyl-L-prolyl alcohol test sample, precisely weighing, and preparing 224mg of N-methyl-L-prolyl alcohol test sample per 1mL of test sample solution by using 100% methanol.
Method for detecting N-methyl-L-prolyl related substances
The detection analysis was performed according to the following chromatographic conditions:
chromatographic column: agilent DB-1701 30m X0.32 mm,0.25 μm;
carrier gas flow rate: 1.0mL/min
Split ratio: 50:1
Heating program: the initial temperature is 45 ℃, maintained for 2 minutes, and the temperature is raised to 280 ℃ at the rate of 20 ℃ per minute, and maintained for 5 minutes;
sample inlet temperature: 280 DEG C
Detector temperature: 280 DEG C
And (3) methodological verification:
(1) Specificity and System applicability test
Taking a blank solvent, a system applicability solution and a sample solution to be detected according to the gas chromatography conditions, detecting the sample injection amount by 1 mu L, and recording a chromatogram, wherein the chromatogram peak of the blank solvent is shown in fig. 1, the chromatogram peak of the system applicability solution is shown in fig. 2, the chromatogram peak of the sample solution is shown in fig. 3, wherein the chromatogram peak 1 of the system applicability solution is N-methyl-L-prolyl alcohol, the peak 2 is L-prolyl alcohol, the peak 3 is N-methyl-L-proline, and the experimental results of the system applicability solution are shown in tables 1 and 2:
table 1: corresponding chromatographic peak results after gas chromatography
Table 2: relative standard deviation results for the applicability solution of the repeat sample injection system
The test result shows that the baseline is stable and has no interference, and the blank solvent has no interference on detection of taking L-prolyl, N-methyl-L-proline and N-methyl-L-prolyl; the N-methyl-L-prolyl theoretical plate number in the system applicability solution is larger than 5000, the separation degree between the N-methyl-L-prolyl and the separation degree between the N-methyl-L-prolyl are larger than 1.5, and the detection method is suitable for detecting related substances of the N-methyl-L-prolyl, so that the detection method provided by the invention has good specificity, the repeated sample injection system is suitable for 6 needles of the solution, the RSD of the peak area of the N-methyl-L-prolyl is 0.87 percent and smaller than 2.0 percent, and meets the standard, and the system applicability is good.
(2) Limit of detection and limit of quantification
The concentration of each impurity at a signal-to-noise ratio of 3:1 was set as the detection limit, and the signal-to-noise ratio was 10: and (3) determining the concentration at the time 1 as a quantitative limit, respectively sampling and detecting according to the chromatographic conditions, continuously sampling the quantitative limit solution for 6 times, and calculating the RSD of the peak area. The quantitative limit and the detection limit test results are shown in Table 3, and the quantitative limit reproducibility test results are shown in Table 4:
table 3: quantitative limit and detection limit test results
Table 4: quantitative limit reproducibility test results
The quantitative limit and the detection limit test result show that the detection method provided by the invention has high sensitivity, and the repeated determination of the peak areas of the L-prolyl and the N-methyl-L-proline for 6 times has RSD less than 10.0 percent, which indicates that the method provided by the scheme has good quantitative limit repeatability.
(3) Linear range
Precisely weighing L-prolyl alcohol and N-methyl-L-proline, diluting with blank solvent to obtain linear solution with serial concentration, and detecting according to the above chromatographic conditions and detection method. 1. Mu.L of each linear solution was precisely measured and injected into a gas chromatograph, the chromatogram was recorded, the peak area was measured, and linear regression was performed with the peak area A as the ordinate and the concentration C as the abscissa, and the results are shown in tables 5 and 6.
Table 5: L-Pronol Linear test results
Table 6: N-methyl-L-proline linearity test results
The test result shows that the components have good linear relation, and the detection method provided by the invention has good linear relation.
(4) Recovery rate
Taking a proper amount of L-prolyl alcohol and N-methyl-L-proline, precisely weighing, preparing a mixed solution with the limit concentration of 10 times by using 100% methanol, taking about 2240mg of an N-methyl-L-prolyl alcohol test sample as an impurity reference substance stock solution, precisely weighing, placing into a 10mL measuring flask, precisely preparing 9 parts, respectively precisely adding 1.6mL, 2mL and 2.4mL of the impurity reference substance stock solution, metering the volume to a scale by using 100% methanol, preparing 3 parts of each concentration in parallel, taking the recovery ratio test sample solution, and detecting according to the chromatographic conditions and the detection method, wherein the recovery ratio results of each component are shown in tables 7 and 8.
Table 7: l-prolyl recovery rate detection result
Table 8: N-methyl-L-proline recovery rate detection result
From the data, the recovery rate of each component is between 80% and 120%, and the recovery rate RSD of the solution with the same concentration is less than 10.0% and is within acceptable standards. Therefore, the detection method provided by the invention is also proved to have good accuracy.
(5) Repeatability of
3 parts of 100% standard solution is taken, detection is carried out according to the chromatographic conditions and the detection method, a chromatogram is recorded, and the content of each related substance in N-methyl-L-prolyl is calculated, and the results are shown in Table 9.
Table 9: results of the repeatability test
The test result shows that the detection results of 3 parts of 100% labeling solution have no obvious difference, and the detection method provided by the invention has good repeatability.
(6) Solution stability
Taking 1 part of 100% standard adding solution, and respectively detecting at 0h, 2h, 4h, 8h, 12h and 24h according to the chromatographic conditions and the detection method, wherein the solution stability results are shown in Table 10:
table 10: results table of solution stability
The test results show that the RSD of the peak areas of the L-prolyl alcohol and the N-methyl-L-proline is less than 10.0%, and the detection method provided by the invention has good solution stability after being placed for 24 hours at room temperature.
(7) Durability of
Taking a plurality of parts of system applicability solution, respectively changing the temperature of a sample inlet, the initial temperature, the temperature rising rate and the temperature of a detector, observing the chromatographic behavior change of an instrument, and the durability results are shown in tables 11 to 14:
table 11: different sample inlet temperature separation degree
| Name of the name | The temperature of the sample inlet is 275 DEG C | The temperature of the sample inlet is 280 DEG C | The temperature of the sample inlet is 285 DEG C |
| N-methyl-L-prolinol | _ | _ | _ |
| L-prolyl alcohol | 4.33 | 4.57 | 4.35 |
| N-methyl-L-proline | 58.78 | 59.18 | 58.49 |
Table 12: degree of separation at different onset temperatures
| Name of the name | Initial temperature 40 DEG C | Initial temperature 45 DEG C | Initial temperature of 50 DEG C |
| N-methyl-L-prolinol | _ | _ | _ |
| L-prolyl alcohol | 4.27 | 4.57 | 4.34 |
| N-methyl-L-proline | 58.58 | 59.18 | 57.81 |
Table 13: degree of separation at different heating rates
Table 14: different detector temperature separation
| Name of the name | Detector 275 deg.c | Detector 280 deg.c | Detector 285 DEG C |
| N-methyl-L-prolinol | _ | _ | _ |
| L-prolyl alcohol | 4.39 | 4.57 | 4.34 |
| N-methyl-L-proline | 58.34 | 59.18 | 58.95 |
The detection method provided by the invention has good durability as various conditions are changed and the separation degree of each component is more than 1.5, which meets the requirements.
(8) Test solution detection
The sample solution was tested under the above chromatographic conditions, and the chromatogram is shown in FIG. 3. The related substances in the test solution are calculated by the external standard method according to the mixed reference solution respectively. The results are shown in Table 15.
Table 15: detection results of related substances of test sample
The data show that the separation degree between the main peak and the adjacent impurities is more than or equal to 1.5, and meets the regulations, which indicates that the N-methyl-L-prolyl alcohol can be effectively separated from the L-prolyl alcohol and the N-methyl-L-proline.
In order to better embody the effectiveness of the method of the present invention, the following comparative analysis of N-methyl-L-prolyl-related substances was conducted under different conditions in comparative examples.
Comparative example 1:
detection was performed by gas chromatography under the following conditions:
chromatographic column: agilent HP-5 30 m.times.0.53 mm,1.50 μm;
carrier gas flow rate: 5.0mL/min;
split ratio: 10:1, a step of;
heating program: the initial temperature is 40 ℃, maintained for 6 minutes, and the temperature is raised to 240 ℃ at the rate of 20 ℃ per minute, and maintained for 10 minutes;
sample inlet temperature: 250 ℃;
detector temperature: 280 DEG C
Dilution liquid: 100% methanol.
The chromatogram is shown in FIG. 4. Under this method, the chromatographic peak shapes of N-methyl-L-prolyl alcohol, L-prolyl alcohol and N-methyl-L-proline are not good, and further optimization is needed.
Comparative example 2:
detection was performed by gas chromatography under the following conditions:
chromatographic column: SE-54 30 mX0.32 mm,0.50 μm;
carrier gas flow rate: 1.0mL/min;
split ratio: 10:1, a step of;
heating program: the initial temperature is 40 ℃, maintained for 6 minutes, and the temperature is raised to 240 ℃ at the rate of 20 ℃ per minute, and maintained for 10 minutes;
sample inlet temperature: 250 ℃;
detector temperature: 280 DEG C
Dilution liquid: 100% methanol.
The chromatograms of the method are shown in figure 5, and the chromatographic peak shapes of the N-methyl-L-prolyl alcohol, the L-prolyl alcohol and the N-methyl-L-proline are not good, so that the method needs further optimization.
Comparative example 3:
detection was performed by gas chromatography under the following conditions:
chromatographic column: agilent HP-FFAP 30m×0.53mm,1.00 μm;
carrier gas flow rate: 3.0mL/min;
split ratio: 5:1, a step of;
heating program: the initial temperature is 60 ℃, maintained for 6 minutes, and the temperature is raised to 230 ℃ at the rate of 20 ℃ per minute, and maintained for 5.5 minutes;
sample inlet temperature: 250 ℃;
detector temperature: 280 DEG C
Dilution liquid: 100% methanol.
The chromatogram is shown in FIG. 6. Under the method, the chromatographic peak shape of the N-methyl-L-prolyl alcohol is poor, the peaks of the L-prolyl alcohol and the N-methyl-L-proline are not generated, and the method is not applicable.
Comparative example 4:
detection was performed by liquid chromatography under the following conditions:
chromatographic column: CAPCELL Pak C18 4.6mm 250mm 5 μm
Mobile phase a:6.0g/L anhydrous sodium dihydrogen phosphate, regulating the pH to 3.0 by phosphoric acid, adding 2.35g/L sodium octane sulfonate, stirring uniformly, and filtering.
Mobile phase B: acetonitrile
Flow rate: 1.0mL/min
A detector: ultraviolet detector (UV)
Wavelength: 200nm
Column temperature: 30 DEG C
The gradient elution procedure was as follows:
| time (min) | Mobile phase A (% V/V) | Mobile phase B (% V/V) |
| 0 | 90 | 10 |
| 30 | 70 | 30 |
| 35 | 90 | 10 |
| 50 | 90 | 10 |
The chromatogram is shown in FIG. 7. The liquid phase condition is selected for screening, and no peak appears in the N-methyl-L-prolyl alcohol, the L-prolyl alcohol and the N-methyl-L-proline, so that the method is not applicable.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, or alternatives falling within the spirit and principles of the invention.
Claims (6)
1. The detection method of the N-methyl-L-prolyl related substances is characterized in that a solution to be detected is prepared by taking 100% methanol as a solvent, and the related substances are detected by using a gas chromatography, wherein the related substances comprise L-prolyl and N-methyl-L-proline, and the chromatographic conditions of the gas chromatography are as follows:
chromatographic column: capillary column with cyanopropyl phenyl-dimethyl polysiloxane as stationary phase;
carrier gas flow rate: 0.5 to 1.5ml/min;
split ratio: 48-52:1;
heating program: the initial temperature is 40-50 ℃, the temperature is maintained for 1-3 minutes, the temperature is raised to 275-285 ℃ at the rate of 18-22 ℃ per minute, and the temperature is maintained for 2-8 minutes;
sample inlet temperature: 275-285 ℃;
the detector is a hydrogen flame ionization detector, and the temperature of the detector is 275-285 ℃.
2. The method for detecting an N-methyl-L-prolinol-related substance according to claim 1, comprising the steps of:
preparing a sample solution, and dissolving N-methyl-L-prolyl alcohol by using a solvent to obtain the sample solution;
preparing a system applicability solution, namely dissolving an N-methyl-L-prolyl reference substance and an impurity reference substance stock solution by using a solvent to obtain the system applicability solution; the impurity reference substance stock solution is prepared by dissolving an impurity reference substance with a solvent, wherein the impurities comprise L-prolyl and N-methyl-L-proline;
detection was performed by gas chromatography under the following conditions:
chromatographic column: agilent DB-1701 30m X0.32 mm,0.25 μm;
carrier gas flow rate: 1.0mL/min;
split ratio: 50:1;
heating program: the initial temperature is 45 ℃, maintained for 2 minutes, and the temperature is raised to 280 ℃ at the rate of 20 ℃ per minute, and maintained for 5 minutes;
sample inlet temperature: 280 ℃;
detector temperature: 280 ℃.
3. The method for detecting an N-methyl-L-prolinol-related substance according to claim 2, further comprising the steps of: and calculating the content of the N-methyl-L-prolyl related substances in the test sample solution according to the measured data by an external standard method and peak areas.
4. The method for detecting an N-methyl-L-prolinol-related substance according to claim 2, wherein the column is a capillary column having 14% cyanopropylphenyl-86% dimethylpolysiloxane as a stationary phase.
5. The method for detecting an N-methyl-L-prolinol-related substance according to claim 1, wherein the limit of L-prolinol is not more than 0.5%.
6. The method for detecting an N-methyl-L-prolinol-related substance according to claim 1, wherein the limit of N-methyl-L-proline is not more than 0.5%.
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