Method for detecting impurities of dextroborneol

Abstract

The invention relates to the technical field of medicine detection, in particular to a method for detecting impurities of dextroborneol. The detection method provided by the invention comprises the following steps: providing a dextrocamphol test solution and a reference substance solution of dextrocamphol impurities, wherein the dextrocamphol impurities comprise at least one of dextrocamphol, isoborneol and L-borneol; carrying out gas chromatography-mass spectrometry detection on the dextrocamphol test solution and the reference substance solution of the dextrocamphol impurity; wherein certain chromatographic conditions and mass spectrum conditions are set. According to the embodiment of the invention, by setting related chromatographic conditions and mass spectrum conditions, the final detection result shows that the peak pattern is good, the separation degree is excellent, the method has obvious advantages in specificity, quantitation limit, detection limit, linearity, precision, accuracy and durability, and a detection thought is provided for impurity detection of dextroborneol.

Description

Technical Field

[0001] The present application belongs to the technical field of drug detection, and in particular, relates to a method for detecting dexamethasone impurities. Background Art

[0002] Dextroborneol, chemical name endo-(1R)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-ol, CAS number 464-43-7, is a natural bicyclic monoterpene used for analgesia and anesthesia. Modern pharmacological research has found that dextroborneol has pharmacological effects such as protecting nerves, increasing blood-brain barrier permeability, shortening sleep time, protecting heart and brain organs and tissues, transdermal absorption, antibacterial, anti-inflammatory, sedative, analgesic, antiseptic and muscle-building, and skin care.

[0003] In recent years, the demand for borneol has been increasing day by day. Due to its high cost of use, synthetic borneol is often used instead. However, in addition to the main chemical component borneol, synthetic borneol also contains a large amount of isoborneol, which has certain toxic and side effects. In addition, synthetic borneol will gradually convert into camphor during storage, and camphor has strong toxicity and can act on the nervous system, causing severe poisoning.

[0004] Given the differences in safety and pharmacokinetic properties of borneol, detecting the content of impurities such as d-camphor, isoborneol and levoborneol is significantly helpful in controlling the quality of borneol. However, the current detection method needs to be improved. Summary of the invention

[0005] The purpose of the present application is to provide a method for detecting impurities in borneol, aiming to solve the technical problem of how to better detect impurities in borneol.

[0006] In order to achieve the above application purpose, the technical solution adopted in this application is as follows:

[0007] The present application provides a method for detecting dextroborneol impurities, comprising:

[0008] Providing a borneol test solution and a borneol reference solution of impurities; the borneol impurities include at least one of dextrorotatory camphor, isoborneol and l-borneol;

[0009] Performing gas chromatography-mass spectrometry detection on the dextroborneol test solution and the dextroborneol impurity reference solution;

[0010] in,

[0011] The chromatographic conditions include: using a chiral capillary column as a chromatographic column, program temperature increase, the initial column temperature is 75-85°C, maintained for 7-9 minutes; heating to 105-115°C at a rate of 4-6°C / min, maintained for 12-14 minutes; then heating to 175-185°C at a rate of 4-6°C / min, and then heating to 215-225°C at a rate of 38-42°C / min, maintained for 4-6 minutes; the injection port temperature is 295-305°C; the flow rate is 1.9-2.1 ml / min, the split ratio is 18-22:1, and the injection volume is 0.5-1.5 μl;

[0012] The mass spectrometry conditions include: the ion source is an electron impact ion source; the ion source temperature is 270-290° C.; the quantitative ion is 95; the electron bombardment energy is 70 eV; and the scanning mode is the SIM mode.

[0013] The method for detecting impurities in dextroborneol provided in the present application uses gas chromatography-mass spectrometry (GCMS) to detect impurities in dextroborneol. By setting relevant chromatographic conditions and mass spectrometry conditions, the final detection results show that the peak shape is good and the separation is excellent. It shows obvious advantages in specificity, quantitative limit, detection limit, linearity, precision, accuracy and durability. It has the advantages of strong specificity, high sensitivity and precision, and good accuracy, and provides a detection idea for the detection of related impurities in the raw materials and preparations of dextroborneol. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0015] Figure 1 is the liquid phase chromatogram of the blank solvent in Example 2 of the present application;

[0016] Figure 2 is the liquid phase mass spectrum of the blank solvent in Example 2 of the present application;

[0017] Figure 3 is the chromatogram of the reference solution in Example 2 of the present application;

[0018] Figure 4 is the mass spectrum of the reference solution in Example 2 of the present application;

[0019] Figure 5 It is the chromatogram of the test solution in Example 2 of the present application;

[0020] Figure 6 This is the mass spectrum of the test solution in Example 2 of the present application. DETAILED DESCRIPTION

[0021] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application more clearly understood, the present application is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0022] In this application, the term "and / or" describes the association relationship of associated objects, indicating that there may be three relationships. For example, A and / or B can mean: A exists alone, A and B exist at the same time, and B exists alone. A and B can be singular or plural. The character " / " generally indicates that the associated objects are in an "or" relationship.

[0023] In the present application, "at least one" means one or more, "more than one" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single or plural items.

[0024] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the above-mentioned processes does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0025] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments, and are not intended to limit the present application. The singular forms "a", "said" and "the" used in the embodiments of the present application and the appended claims are also intended to include plural forms, unless the context clearly indicates other meanings.

[0026] The weight of the relevant components mentioned in the embodiment description of the present application can not only refer to the specific content of each component, but also represent the proportional relationship between the weights of the components. Therefore, as long as the content of the relevant components is proportionally enlarged or reduced according to the embodiment description of the present application, it is within the scope disclosed in the embodiment description of the present application. Specifically, the mass described in the embodiment description of the present application can be a mass unit known in the chemical industry such as μg, mg, g, kg, etc.

[0027] The terms "first" and "second" are used only for descriptive purposes to distinguish objects such as substances from each other, and should not be understood as indicating or implying relative importance or implicitly indicating the number of indicated technical features. For example, without departing from the scope of the embodiments of the present application, the first XX may also be referred to as the second XX, and similarly, the second XX may also be referred to as the first XX. Thus, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features.

[0028] Borneol is the main active ingredient of Camphor Camphor, also known as natural borneol. It is a commonly used precious Chinese medicinal material and high-grade edible spice, which has the effects of refreshing the mind, clearing away heat and relieving pain. In recent years, the demand for Borneol has increased day by day. Due to its high cost of use, it is often replaced by synthetic Borneol. However, in addition to the main chemical component D-borneol, synthetic Borneol also contains a large amount of isoborneol, which has certain toxic and side effects. In addition, synthetic Borneol will gradually convert into camphor during storage, and camphor is highly toxic and can act on the nervous system, causing severe poisoning. In view of the differences in safety and pharmacokinetic properties of Borneol, detecting the content of impurities such as D-camphor, isoborneol and L-borneol is significantly helpful for controlling the quality of Borneol. The relevant molecular structure information is shown in Table 1.

[0029] Table 1

[0030]

[0031] At present, the Chinese Pharmacopoeia does not include varieties that simultaneously detect the content of impurities such as right-handed camphor, isoborneol and left-handed camphor. In the existing public related literature, there are impurity detections of some different chiral isomers, and the commonly used methods are single gas chromatography (GC) or high performance liquid chromatography (HPLC) detection. These methods can only be quantitative, and can only be qualitative by retention time, and the latter detection baseline is prone to fluctuations, and the detection effect is not ideal.

[0032] To this end, the present application embodiment has developed a method for detecting impurities in borneol (such as at least one borneol impurity in d-camphor, isoborneol and levorotatory borneol, etc.) by GCMS method after comprehensive and systematic methodological research, which has the advantages of strong specificity, high sensitivity, good precision and high accuracy. The technical solution is described in detail below.

[0033] The present application embodiment provides a method for detecting borneol impurities. The detection method provided in the present application embodiment includes:

[0034] S01: providing a borneol test solution and a reference solution of borneol impurities; wherein the borneol impurities include at least one of dextrorotatory camphor, isoborneol and l-borneol;

[0035] S02: gas chromatography-mass spectrometry detection is carried out to the reference substance solution of dextroborneol need testing solution and dextroborneol impurity;

[0036] in,

[0037] The chromatographic conditions include: using a chiral capillary column as a chromatographic column, program temperature increase, the initial column temperature is 75-85°C, maintained for 7-9 minutes; heating to 105-115°C at a rate of 4-6°C / min, maintained for 12-14 minutes; then heating to 175-185°C at a rate of 4-6°C / min, and then heating to 215-225°C at a rate of 38-42°C / min, maintained for 4-6 minutes; the injection port temperature is 295-305°C; the flow rate is 1.9-2.1 ml / min, the split ratio is 18-22:1, and the injection volume is 0.5-1.5 μl;

[0038] The mass spectrometry conditions include: the ion source is an electron impact ion source; the ion source temperature is 270-290° C.; the quantitative ion is 95; the electron bombardment energy is 70 eV; and the scanning mode is the SIM mode.

[0039] The purpose of the embodiments of the present application is to provide a detection method based on the GCMS system with higher detection precision, shorter detection time, and better system applicability, so that it has advantages in specificity, quantitative limit, detection limit, linear range, repeatability, accuracy, etc., and provides an important method detection reference basis for the quality control of dextromethorphan drug solution or dextromethorphan-containing preparation solution (such as: edaravone dextromethorphan concentrated solution for injection).

[0040] Further, the chiral capillary column in the chromatographic conditions includes a Cyclosil-B chromatographic column. For example, the specifications of the Cyclosil-B chromatographic column may be: 30m×0.25mm, 0.25μm.

[0041] In some embodiments, the chromatographic conditions include: using Cyclosil-B as the chromatographic column, programmed temperature, the initial column temperature is 80°C, maintained for 8 minutes; heating to 110°C at a rate of 5°C / min, maintained for 13 minutes; then heating to 180°C at a rate of 5°C / min, and then heating to 220°C at a rate of 40°C / min, maintained for 5 minutes; the carrier gas is helium; the injection port temperature is 300°C; the flow rate is 2.0 ml / min, the split ratio is 20:1, and the injection volume is 1 μl.

[0042] In some embodiments, the mass spectrometry conditions include: the ion source is an electron impact ion (EI) source; the ion source temperature is 280° C.; the quantitative ion is 95; the electron bombardment energy is 70 eV; and the scanning mode is selected as SIM mode.

[0043] In some embodiments, the dextroborneol test solution is a solution containing dextroborneol to be tested, and specifically may include a dextroborneol bulk drug solution or a dextroborneol-containing preparation solution. For a preparation solution containing dextroborneol, a commonly used pharmaceutical preparation containing dextroborneol such as edaravone dextroborneol injection concentrated solution may be used. According to the present application, it is a detection method, which can mainly be directed to the detection of dextroborneol raw materials and preparation products (such as: edaravone dextroborneol injection concentrated solution).

[0044] In some embodiments, the solvent in the dextroborneol test solution is selected from methanol. For example, taking the dextroborneol test solution as an edaravone dextroborneol concentrated solution for injection as an example, the test solution preparation includes: accurately measuring 5 ml of the edaravone dextroborneol concentrated solution for injection (compound injection), placing it in a 10 ml volumetric bottle, adding methanol to dissolve and dilute to the scale, shaking well, and using it as the test solution.

[0045] In some embodiments, the standard curve method of the reference substance is used to calculate the dextroborneol impurity in the dextroborneol test solution. Wherein, the solvent in the reference substance solution of the dextroborneol impurity is selected from a methanol aqueous solution with a methanol volume percentage of 45 to 55%.

[0046] Specifically, the reference substance stock solution can be prepared: take right-handed camphor, isoborneol and left-handed camphor, weigh accurately, put into a volumetric bottle, add methanol to dissolve and dilute to scale, shake well; further, add 45-55% methanol solution to dilute to scale, shake well, as the reference substance stock solution. Then take the reference substance stock solution, add 45-55% methanol solution to dilute to prepare a series of reference substance standard curve solutions of different concentrations.

[0047] Furthermore, a system suitability solution can be prepared. For example, taking the dextroborneol test solution as an edaravone dextroborneol concentrated solution for injection as an example: accurately measure 1 ml of the edaravone dextroborneol concentrated solution for injection, place it in a 10 ml volumetric bottle, add 1 ml of the above-mentioned reference substance stock solution, add methanol to dilute to the scale, shake well, and use it as the system suitability solution.

[0048] In certain embodiments, according to the detection method of impurities in right-borneol, the quality control limit of right-handed camphor is 0.2%, the quality control limit of isoborneol is 0.3%, and the quality control limit of left-handed camphor is 1.0%. In the quantitative limit and detection limit test, the impurity quantitative limit concentration and detection limit concentration of right-borneol are both lower than 1 / 10 of its quality control limit; when the detection method based on GCMS of the present application embodiment is used for detection, the detection limit of right-handed camphor is 0.006%, the detection limit of isoborneol is 0.009%, and the detection limit of left-handed camphor is 0.03%, indicating that right-handed camphor, isoborneol and left-handed camphor can be effectively detected above the quality control limit of 3 / 100.

[0049] In some embodiments, when the GCMS-based detection method of the embodiment of the present application is used for detection, a system suitability solution containing right-handed camphor, isoborneol, left-handed camphor and right-borneol, in a specific test, in the chromatogram of the system suitability solution, right-handed camphor, isoborneol-1, isoborneol-2, left-handed camphor, and right-borneol are eluted in sequence, and the separation between the left-handed camphor peak and the right-borneol peak should be greater than 1.5, and the number of theoretical plates should be not less than 3000 based on left-handed camphor.

[0050] In certain embodiments, in the system suitability test, 6 injections were continuously made, and the RSD% of the peak areas of right-handed camphor, isoborneol and left-handed camphor were all less than 8.0%. In the test of linear range, the linear relationship of right-handed camphor, isoborneol and left-handed camphor was good within the range of (10% to 150%) relative to the test sample concentration, and the correlation coefficient r was not less than 0.99. In the solution stability test, within 24 hours at room temperature, the RSD% of the peak areas of right-handed camphor, isoborneol and left-handed camphor in the reference solution and the test sample solution was all less than 8.0%. In the test of accuracy, the recovery of right-handed camphor, isoborneol and left-handed camphor was between 80% and 120%, and the RSD% was all less than 8.0%. In the test of repeatability, the RSD% of the detection results of right-handed camphor, isoborneol and left-handed camphor in 6 samples was all less than 8.0%. In the intermediate precision test, the RSD% of the detection results of right-rotating camphor, isoborneol and left-rotating borneol in 12 samples were all less than 8.0%.

[0051] Therefore, the present application embodiment adopts the detection method of the impurities (at least one impurity such as d-camphor, isoborneol and borneol) of d-camphor based on GCMS detection. The final result shows: the peak shape is good, the separation is excellent, and it shows obvious advantages in specificity, quantitative limit, detection limit, linearity, precision, accuracy and durability, and has the advantages of strong specificity, high precision and high accuracy, and provides detection ideas for the raw materials and preparation products of d-camphor, isoborneol and borneol.

[0052] For the detection method of the impurities (at least one impurity such as dextrorotatory camphor, isoborneol and borneol) of borneol detected based on the GCMS system provided in the embodiment of the present application, those skilled in the art can learn from the content of this article and appropriately improve the chromatogram and mass spectrometry parameters to achieve. It is particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all deemed to be included in the present application. The present application detection method is described by preferred embodiments, and the relevant personnel obviously change or appropriately change and combine the method application described herein without departing from the content, spirit and scope of the present application to realize and apply the present application technology.

[0053] The present application is further described below in conjunction with the examples. The gas chromatograph-mass spectrometer used in the detection method of the present application in the embodiment is a Thermo Fisher brand GCMS.

[0054] Example 1 Solution preparation and gas chromatography-mass spectrometry conditions

[0055] Preparation of test solution: Accurately measure 5 ml of edaravone and dextroborneol concentrated solution for injection (compound injection, dextroborneol is a main component of the preparation), place it in a 10 ml volumetric flask, add methanol to dilute to the scale (10 ml), shake well, and use it as the test solution.

[0056] Preparation of reference substance stock solution: Take 5 mg of right-handed camphor, 7.5 mg of isoborneol and 25 mg of left-handed camphor, accurately weigh them, place them in 3 different 10 ml volumetric flasks, add methanol to dissolve and dilute to the scale, shake well, and use them as the reference substance intermediate stock solution; accurately measure 1 ml, place it in a 100 ml volumetric flask, add 50% methanol aqueous solution to dilute to the scale, shake well, and use it as the reference substance stock solution.

[0057] Preparation of reference substance standard curve solution: Take the above reference substance stock solution and dilute it with 50% methanol aqueous solution to prepare a series of reference substance solutions containing 50 ng, 100 ng, 200 ng, 350 ng, 500 ng and 750 ng of dextrorotatory camphor per 1 ml (the reference substance standard curve solutions are named L1, L2, L3, L4, L5 and L6, respectively).

[0058] System suitability solution: Accurately measure the preparation solution containing dexborneol: 5 ml of edaravone dexborneol concentrated solution for injection, place it in a 10 ml volumetric flask, add 1.0 ml of the above-mentioned reference substance stock solution, add methanol to dilute to the scale, shake well, and use it as the system suitability solution.

[0059] Chromatographic conditions: capillary column chromatography column (Cyclosil-B, 30m*0.25mm*0.25μm) with β-cyclodextrin as the stationary liquid, programmed temperature, initial column temperature is 80℃, maintained for 8 minutes; heated to 110℃ at a rate of 5℃ per minute, maintained for 13 minutes; then heated to 180℃ at a rate of 5℃ per minute; heated to 220℃ at a rate of 40℃ per minute, maintained for 5 minutes; carrier gas is helium; injection port temperature is 300℃; flow rate is 2.0ml / min, split ratio is 20:1, injection volume is 1μl.

[0060] Mass spectrometry conditions: ion source: EI source; ion source temperature: 280°C; quantitative ion: 95; electron bombardment energy: 70 eV; scanning mode: SIM mode.

[0061] Example 2 Specificity Test

[0062] Solvent: methanol.

[0063] Blank solvent: 50% methanol in water.

[0064] System suitability solution: Accurately measure 5 ml of the concentrated solution of edaravone and dextroborneol for injection, place it in a 10 ml volumetric flask, add 1.0 ml of the reference substance stock solution of Example 1 above, dilute to the scale with methanol, shake well, and use as the system suitability solution.

[0065] Test solution: Accurately measure 5 ml of edaravone and dextroborneol concentrated solution for injection into a 10 ml volumetric flask, add methanol to dissolve and dilute to the scale, shake well, and use as the test solution.

[0066] The above system suitability solution and the test solution were subjected to gas chromatography-mass spectrometry detection (chromatographic conditions and mass spectrometry conditions were the same as those in Example 1). The specificity test results are shown in Table 2 and the spectra are shown in Attachment Figure 1 ~Attached Figure 6 Conclusion: The blank solvent does not interfere with the detection of right-rotating camphor, isoborneol and left-rotating camphor. Right-rotating camphor, isoborneol and left-rotating camphor can be effectively separated from right-borneol with a separation degree of >1.5. The detection method has good specificity.

[0067] Table 2

[0068] Serial number Peak name Retention time Separation Theoretical plates 1 Dextrorotatory camphor 18.756 34.09 325400 2 Isoborneol-1 24.546 3.49 220533 3 Isoborneol-2 25.281 2.44 224158 4 L-Borneol 25.806 3.70 224443 5 Dextroborneol 26.821 / 104169

[0069] Note: There are two peaks of isoborneol: isoborneol-1 and isoborneol-2.

[0070] Example 3 System suitability test of the detection method

[0071] The reference solution was prepared according to the preparation method of the reference standard curve solution L5 in Example 1 above, 1 μl of the reference solution was accurately drawn, injected into a gas chromatography-mass spectrometer, and gas chromatography-mass spectrometry detection was performed (chromatographic conditions and mass spectrometry conditions were the same as in Example 1), and a chromatogram was recorded. The system suitability results are shown in Table 3. The results showed that the reference solution was injected 6 times continuously, and the RSD of right-handed camphor was 4.3%, the RSD of isoborneol was 3.8%, and the RSD of left-handed borneol was 3.7%, all of which were less than 8.0%, and the detection method had good system suitability.

[0072] Table 3

[0073] Components 6-needle peak area Average peak area RSD(%) Dextrorotatory camphor 2401、2260、2325、2183、2306、2465 2323 4.3 Isoborneol 7952、7682、7675、7244、8016、7979 7758 3.8 L-Borneol 29960、28758、29514、27545、29978、30556 29385 3.7

[0074] Example 4 Linearity and range detection of detection method

[0075] Preparation of reference stock solution: Take 5 mg of right-handed camphor, 7.5 mg of isoborneol and 25 mg of left-handed camphor, accurately weigh them, place them in 3 different 10 ml volumetric flasks, add methanol to dissolve and dilute to the scale, shake well; accurately measure 1 ml, place it in a 100 ml volumetric flask, add 50% methanol aqueous solution to dilute to the scale, shake well, as the reference stock solution.

[0076] Preparation of reference standard curve solution: Prepare L1, L2, L3, L4, L5, and L6 according to the method of Example 1, as shown in Table 4.

[0077] Table 4

[0078]

[0079] Accurately pipette 1 μl of the standard curve solution of the reference substance, inject it into the gas chromatography-mass spectrometry instrument, perform gas chromatography-mass spectrometry detection (chromatographic conditions and mass spectrometry conditions are the same as those in Example 1), and record the chromatogram. Perform linear regression on the peak area based on the concentration of the test solution, and the results are as follows:

[0080] The results of the linear test of right-handed camphor are shown in Table 5. The results show that right-handed camphor presents a good linear relationship in the concentration range of 50 to 750 ng / ml, and the correlation coefficient r is not less than 0.995; the Y-axis intercept deviation is 0.3%, which meets the requirement that the intercept deviation shall not exceed 25.0%, and the linearity is good.

[0081] Table 5

[0082]

[0083] The results of the linear test of isoborneol are shown in Table 6. The results show that isoborneol presents a good linear relationship in the concentration range of 75 to 1125 ng / ml, the correlation coefficient r is not less than 0.995, the Y-axis intercept deviation is 0.01%, which meets the requirement that the intercept deviation shall not exceed 25.0%, and the linearity is good.

[0084] Table 6

[0085]

[0086] The linear test results of L-borneol are shown in Table 7, and the results show that: the results show that L-borneol presents a good linear relationship in the concentration range of 250-3750 ng / ml, the correlation coefficient r is not less than 0.995, and the Y-axis intercept deviation is 0.1%, which meets the requirement that the intercept deviation shall not exceed 25.0%, and the linearity is good.

[0087] Table 7

[0088]

[0089] Example 5 Quantitative Limit and Detection Limit Test of Detection Method

[0090] Take the reference standard curve solutions L5 and L4 in Table 4 of Example 4, add 50% methanol aqueous solution as solvent, and dilute according to the following Table 8.

[0091] Table 8

[0092]

[0093] Dextrorotatory camphor concentration is 50ng / ml, and signal-to-noise ratio S / N is 11.9, isoborneol concentration is 75ng / ml, peak 1 signal-to-noise ratio S / N is 17.0, peak 2 signal-to-noise ratio S / N is 13.6, and L-borneol concentration is 250ng / ml, and signal-to-noise ratio S / N is 673.1, which meets the requirement of 10 times of signal-to-noise ratio and is proposed as quantitative limit. Continuous injection 6 needles, carry out gas chromatography-mass spectrometry detection (chromatographic conditions and mass spectrometry conditions are the same as embodiment 1), peak area and RSD results are as shown in table 9. The result shows: quantitative limit solution is continuously injected 6 needles, and dextrorotatory camphor RSD is 3.7%, isoborneol RSD is 6.5%, and L-borneol RSD is 3.7%, which is less than 8.0%, meets the requirements.

[0094] Table 9

[0095]

[0096] The concentration of right-handed camphor is 15 ng / ml, the signal-to-noise ratio S / N is 3.8, the concentration of isoborneol is 22.5 ng / ml, the signal-to-noise ratio S / N of peak 1 is 6.2, the signal-to-noise ratio S / N of peak 2 is 5.8, the concentration of left-handed camphor is 75 ng / ml, the signal-to-noise ratio S / N is 359.8, which meets the requirement of 3 times the signal-to-noise ratio and is intended to be the detection limit. The results are shown in Table 10 below. The results show that the quality control limit of right-handed camphor is 0.2%, the quality control limit of isoborneol is 0.3%, and the quality control limit of left-handed camphor is 1.0%.

[0097] Table 10

[0098]

[0099] In the quantitative limit and detection limit tests, the quantitative limit concentration and detection limit concentration of the impurity of dextrocamphor were both lower than 1 / 10 of its quality control limit; the detection limit of dextrocamphor was 0.006%, the detection limit of isocamphor was 0.009%, and the detection limit of l-camphor was 0.03%, indicating that dextrocamphor, isocamphor and l-camphor can be effectively detected above the quality control limit of 3 / 100. This method has high sensitivity.

[0100] Example 6 Solution Stability Study of Detection Method

[0101] Test solution: Accurately measure 5 ml of edaravone and dextroborneol concentrated solution for injection into a 10 ml volumetric flask, add methanol to dissolve and dilute to the scale, shake well, and use as the test solution.

[0102] Reference solution: Prepare the reference solution according to the preparation method of reference standard curve solution L5 in Example 1.

[0103] Take the above solution at room temperature and inject at 0h, 4h, 8h, and 24h respectively, perform gas chromatography-mass spectrometry detection (chromatographic conditions and mass spectrometry conditions are the same as those in Example 1), and record the chromatogram. The results are shown in Table 11. The results show that the RSD% of the peak area changes of right-handed camphor, isoborneol, and left-handed camphor in the reference substance and the test sample are all less than 8.0%, and the right-handed camphor, isoborneol, and left-handed camphor in the solution are stable within 24 hours. That is, the detection method solution stability of the present application embodiment is good.

[0104] Table 11

[0105] Time (hours) 0 4 8 24 RSD% Peak area of ​​right-rotatory camphor in the test sample 580 574 534 569 3.4 Peak area of ​​d-camphor in the reference substance 1737 1763 1761 1816 1.9 The peak area of ​​isoborneol in the test sample 1003 1000 877 884 7.4 The peak area of ​​isoborneol in the reference substance 6095 6137 6138 6323 1.6 Peak area of ​​L-borneol in the test sample 13672 13599 12089 11813 7.7 Peak area of ​​L-borneol in reference substance 18524 18878 18877 19287 1.7

[0106] Example 7 Detection of the Repeatability of the Detection Method

[0107] Blank solvent: methanol-water (50:50, volume ratio).

[0108] Test solution: Accurately measure 5 ml of edaravone and dextroborneol concentrated solution for injection into a 10 ml volumetric flask, add methanol to dissolve and dilute to the mark, shake well, and use as the test solution. Prepare 6 portions in parallel.

[0109] Accurately pipette 1 μl of each of the above sample solutions, inject into the gas chromatography-mass spectrometry instrument, perform gas chromatography-mass spectrometry detection (chromatographic conditions and mass spectrometry conditions are the same as those in Example 1), and record the spectrum. The results are shown in Table 12. The results show that in the repeatability test, the RSD% of the detection results of right-handed camphor, isoborneol and left-handed borneol in 6 samples is less than 8.0%, so the method has good repeatability.

[0110] Table 12

[0111]

[0112]

[0113] Example 8 Intermediate precision test of the detection method

[0114] The test solution (the test solution of Example 7) was prepared by different analysts on different dates according to the method under the repeatability item, and 6 portions were prepared in parallel. Injected into a gas chromatography-mass spectrometer, and subjected to gas chromatography-mass spectrometry detection (chromatographic conditions and mass spectrometry conditions were the same as those in Example 1). The results are shown in Table 13. The results show that in the intermediate precision test, the RSD% of the detection results of right-handed camphor, isoborneol and left-handed borneol in 12 samples were all less than 8.0%, and the intermediate precision of the detection method of the embodiment of the present application was good.

[0115] Table 13 Intermediate precision test results

[0116]

[0117] Example 9 Testing of the Accuracy of the Detection Method

[0118] Preparation of accuracy solution: Take the concentrated solution of the test sample edaravone dextroborneol injection and place it in a 20ml volumetric flask, add 80%, 100%, and 120% of the limit of the reference substance intermediate reserve solution in Example 1 to the test sample and dilute to the scale, shake well. Then take 5ml from it and place it in a 10ml volumetric flask, add methanol to dissolve and dilute to the scale, shake well, as the accuracy solution, as shown in Table 14. Prepare 3 portions for each concentration, and prepare 9 portions of accuracy solution in parallel. Then inject it into a gas chromatography-mass spectrometry instrument for gas chromatography-mass spectrometry detection (chromatographic conditions and mass spectrometry conditions are the same as those in Example 1).

[0119] The results of the accuracy test of right-handed camphor are as shown in Table 15, and in the test of accuracy, the recovery of right-handed camphor is between 80%~120%, and RSD% is all less than 8.0%, and accuracy is good. The results of the accuracy test of isoborneol are as shown in Table 16, and in the test of accuracy, the recovery of isoborneol is between 80%~120%, and RSD% is all less than 8.0%, and accuracy is good. The results of the accuracy test of left-borneol are as shown in Table 17, and in the test of accuracy, the recovery of left-borneol is between 80%~120%, and RSD% is all less than 8.0%, and accuracy is good.

[0120] Table 14 Accuracy solution preparation

[0121]

[0122] Table 15 Dextrorotatory camphor accuracy test results

[0123]

[0124] Table 16 Isoborneol accuracy test results

[0125]

[0126] Table 17 L-Borneol accuracy test results

[0127]

[0128] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A method for detecting impurities in dextroborneol, characterized in that, include: Providing a test solution of dextroborneol and a reference solution of dextroborneol impurities; The dextroborneol impurities include at least one of dextrorotatory camphor, isoborneol and l-borneol; Performing gas chromatography-mass spectrometry detection on the dextroborneol test solution and the dextroborneol impurity reference solution; in, The chromatographic conditions include: using a chiral capillary column as a chromatographic column, program temperature increase, the initial column temperature is 75-85°C, maintained for 7-9 minutes; heating to 105-115°C at a rate of 4-6°C / min, maintained for 12-14 minutes; then heating to 175-185°C at a rate of 4-6°C / min, and then heating to 215-225°C at a rate of 38-42°C / min, maintained for 4-6 minutes; the injection port temperature is 295-305°C; the flow rate is 1.9-2.1 ml / min, the split ratio is 18-22:1, and the injection volume is 0.5-1.5 μl; The mass spectrometry conditions include: the ion source is an electron impact ion source; the ion source temperature is 270-290° C.; the quantitative ion is 95; the electron bombardment energy is 70 eV; and the scanning mode is the SIM mode.

2. The detection method according to claim 1, characterized in that The chromatographic conditions include: the initial column temperature is 80°C, maintained for 8 minutes; the temperature is increased to 110°C at a rate of 5°C / min and maintained for 13 minutes; the temperature is then increased to 180°C at a rate of 5°C / min, and then increased to 220°C at a rate of 40°C / min and maintained for 5 minutes; the carrier gas is helium; the injection port temperature is 300°C; the flow rate is 2.0ml / min, the split ratio is 20:1, and the injection volume is 1μl.

3. The detection method according to claim 1, characterized in that The chiral capillary column comprises a Cyclosil-B chromatographic column.

4. The detection method according to claim 1, characterized in that The ion source temperature in the mass spectrometry conditions was 280°C.

5. The detection method according to any one of claims 1 to 4, characterized in that: The dextroborneol impurities include dextrorotatory camphor, isoborneol and l-borneol.

6. The detection method according to claim 5, characterized in that The quality control limit of the right-handed camphor is 0.2%, the quality control limit of the isoborneol is 0.3%, and the quality control limit of the left-handed camphor is 1.0%; the detection limit of the right-handed camphor is 0.006%, the detection limit of the isoborneol is 0.009%, and the detection limit of the left-handed camphor is 0.03%.

7. The detection method according to any one of claims 1 to 4, characterized in that: The solvent in the dextroborneol test solution is selected from methanol.

8. The detection method according to any one of claims 1 to 4, characterized in that: The solvent in the reference solution of the dextroborneol impurity is selected from a methanol aqueous solution with a volume percentage of 45 to 55%.

9. The detection method according to any one of claims 1 to 4, characterized in that: The borneol impurities in the borneol test solution were calculated using a reference standard curve method.

10. The detection method according to any one of claims 1 to 4, characterized in that: The dextroborneol test solution includes a dextroborneol bulk drug solution or a dextroborneol-containing preparation solution.

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