CN116183758B - Analysis method for isomer impurities in diethyl indenamine hydrochloride - Google Patents
Analysis method for isomer impurities in diethyl indenamine hydrochloride Download PDFInfo
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- 239000012535 impurity Substances 0.000 title claims abstract description 103
- 238000004458 analytical method Methods 0.000 title claims abstract description 12
- -1 diethyl indenamine hydrochloride Chemical compound 0.000 title claims description 14
- 238000001514 detection method Methods 0.000 claims abstract description 26
- 238000000926 separation method Methods 0.000 claims abstract description 26
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 16
- WIOQOIHRSCDJIQ-UHFFFAOYSA-N 1,2-diethyl-1h-indene Chemical compound C1=CC=C2C(CC)C(CC)=CC2=C1 WIOQOIHRSCDJIQ-UHFFFAOYSA-N 0.000 claims abstract description 13
- 150000001412 amines Chemical class 0.000 claims abstract description 13
- 238000004817 gas chromatography Methods 0.000 claims abstract description 10
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000001257 hydrogen Substances 0.000 claims abstract description 5
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 5
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical group [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 39
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical group ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 36
- 239000003085 diluting agent Substances 0.000 claims description 34
- 239000000523 sample Substances 0.000 claims description 33
- 239000000243 solution Substances 0.000 claims description 33
- 238000000034 method Methods 0.000 claims description 22
- 238000007865 diluting Methods 0.000 claims description 18
- 238000002347 injection Methods 0.000 claims description 15
- 239000007924 injection Substances 0.000 claims description 15
- 239000011259 mixed solution Substances 0.000 claims description 15
- 238000005303 weighing Methods 0.000 claims description 15
- 239000000047 product Substances 0.000 claims description 14
- 239000012488 sample solution Substances 0.000 claims description 14
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 10
- 239000013558 reference substance Substances 0.000 claims description 10
- 239000012467 final product Substances 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 7
- 238000004090 dissolution Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 239000011550 stock solution Substances 0.000 claims description 7
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical group [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 6
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- 239000002274 desiccant Substances 0.000 claims description 5
- 239000004205 dimethyl polysiloxane Substances 0.000 claims description 5
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims description 5
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims description 5
- 238000005464 sample preparation method Methods 0.000 claims description 5
- 238000005070 sampling Methods 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 239000012085 test solution Substances 0.000 claims description 5
- 230000014759 maintenance of location Effects 0.000 claims description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 2
- 229920001296 polysiloxane Polymers 0.000 claims description 2
- 238000011002 quantification Methods 0.000 abstract description 2
- 235000011121 sodium hydroxide Nutrition 0.000 description 9
- 238000012360 testing method Methods 0.000 description 8
- 239000000126 substance Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 6
- 238000011084 recovery Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- XUKUURHRXDUEBC-SXOMAYOGSA-N (3s,5r)-7-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyheptanoic acid Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-SXOMAYOGSA-N 0.000 description 1
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 230000001088 anti-asthma Effects 0.000 description 1
- 239000000924 antiasthmatic agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940124630 bronchodilator Drugs 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000012458 free base Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 229960004735 indacaterol maleate Drugs 0.000 description 1
- IREJFXIHXRZFER-PCBAQXHCSA-N indacaterol maleate Chemical compound OC(=O)\C=C/C(O)=O.N1C(=O)C=CC2=C1C(O)=CC=C2[C@@H](O)CNC1CC(C=C(C(=C2)CC)CC)=C2C1 IREJFXIHXRZFER-PCBAQXHCSA-N 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000011418 maintenance treatment Methods 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- PQCWTNQXCXUEAO-UHFFFAOYSA-N n,n-diethylaniline;hydron;chloride Chemical compound Cl.CCN(CC)C1=CC=CC=C1 PQCWTNQXCXUEAO-UHFFFAOYSA-N 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 239000011736 potassium bicarbonate Substances 0.000 description 1
- 235000015497 potassium bicarbonate Nutrition 0.000 description 1
- 229910000028 potassium bicarbonate Inorganic materials 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 235000011181 potassium carbonates Nutrition 0.000 description 1
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 235000011118 potassium hydroxide Nutrition 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
Classifications
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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
-
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
-
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/025—Gas chromatography
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
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- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
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Abstract
The invention discloses an analysis method of isomer impurities in diethyl indene amine hydrochloride, which adopts gas chromatography for detection, and the detection conditions are as follows: the chromatographic column is a capillary column; the initial temperature of the column temperature is 55555 ℃, the temperature is raised to 8225822 ℃ at the rate of 8518 ℃ per minute, and the temperature is maintained for 5 minutes; column flow 2.851.5ml/min; sample volume 1 μl; a detector: a hydrogen flame ionization detector; detecting the temperature of 8825222 ℃; the temperature of the sample inlet is 8225822 ℃; the split ratio is 5:1512:1. The invention provides a gas chromatography detection scheme for analyzing isomer impurities of diethyl indene amine hydrochloride, and the detection accuracy and precision are high; under the chromatographic condition, the invention can realize good separation between impurities and between impurity peaks and main component chromatographic peaks, has good peak shape and stable base line, and can realize accurate quantification.
Description
Technical Field
The invention relates to the technical field of impurity detection methods, in particular to an analysis method of isomer impurities in diethyl indene amine hydrochloride.
Background
Indanterol maleate is an antiasthmatic (bronchodilator), is clinically mainly applicable to maintenance treatment of adult Chronic Obstructive Pulmonary Disease (COPD) patients, and is diethyl indenamine hydrochloride serving as a common starting material of indacaterol maleate, and has the following structural formula:
the compound is easy to introduce three structural isomer impurities in the preparation process, and quantitative research on the impurities is needed:
the diethyl indene amine hydrochloride has high boiling point (285.2 ℃), contains a benzene ring structure and has ultraviolet absorption, and related substances are generally controlled by adopting an HPLC method. The method for checking related substances of the material manufacturer comprises the following steps:
chromatographic column: c18, 4.6X105 mm,5 μm;
mobile phase a:0.02mol/L sodium dihydrogen phosphate solution, and regulating the pH value to 3.0 by phosphoric acid;
mobile phase B: methanol;
the gradient elution procedure was as follows:
| time (min) | Mobile phase a (%) | Mobile phase B (%) |
| 0 | 80 | 20 |
| 3 | 80 | 20 |
| 10 | 70 | 30 |
| 20 | 70 | 30 |
| 50 | 20 | 80 |
| 51 | 80 | 20 |
| 60 | 80 | 20 |
。
Flow rate: 1.0ml/min, wavelength: 220nm, column temperature: sample injection volume 10 μl at 40deg.C;
a diluent: methanol water=1:1.
Sample preparation:
impurity A, B, C stock: weighing about 7.5mg of each impurity reference substance, placing into a 50ml measuring flask, adding diluent to dissolve and dilute to scale, and shaking uniformly to obtain the final product.
Mixing solution: measuring 5ml of each impurity A, B, C stock solution, placing into 50ml measuring flask with the concentration of about 25mg, adding diluent, dissolving, diluting to scale, and shaking.
Taking the mixed solution for sample injection, and examining the separation condition of the main peak and the three structural isomer impurity peaks.
As shown in the test result in FIG. 1, according to the detection method of related substances of diethyl indene amine hydrochloride, the main peak and the impurity peaks of three structural isomers cannot be completely separated, the impurity B and the main peak are completely overlapped, and the three structural isomers cannot be accurately quantified by the existing detection method of related substances. As can be seen from the figure, the main peak retention time has been delayed to 14 minutes, and even by adjusting the mobile phase ratio, separation of the three structural isomer impurities from the main component by HPLC method is difficult to achieve, and the analysis work efficiency is affected by too long time. No literature has been searched until any analysis means can control the complete separation and examination of the four compounds.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides an analysis method of isomer impurities in diethyl indene amine hydrochloride; the chromatographic peaks between the three structural isomer impurities and the four compounds of the main component are completely separated by adopting a gas chromatography, and the method has high accuracy and precision, simple analysis operation and high detection efficiency. The technical scheme of the invention is as follows:
an analysis method of isomer impurities in diethyl indene amine hydrochloride adopts gas chromatography for detection, and a chromatographic column adopted is a capillary column;
when the separation degree of each component is more than 1.05, the retention time of the main component is more than 11.3min;
the isomer impurity in the diethyl indene amine hydrochloride is one or more than one of the following structural formula impurities:
the method can realize good separation of the main component from three single impurities respectively, and can also realize good separation between the impurities and the chromatographic peaks of the main component when the three impurities exist simultaneously.
Preferably, the gas chromatography detection conditions are as follows:
column temperature: the initial temperature is 54-66 ℃, the temperature is raised to 230-270 ℃ at the rate of 8-12 ℃ per minute, and the temperature is maintained for at least 5 minutes;
column flow rate: 0.8-1.4 ml/min;
sample injection volume: 1 μl; a detector: a hydrogen flame ionization detector (FID detector);
detecting the temperature: 280-300 ℃; sample inlet temperature: 230-270 ℃; split ratio: 5:1-10:1.
Preferably, the capillary column uses 100% dimethyl polysiloxane, (5%) phenyl- (95%) methyl polysiloxane or other polar similar substances as a fixing liquid.
Further, the capillary column uses 100% dimethyl polysiloxane as a fixing liquid. Preferably, the sample preparation and detection method comprises the following steps:
impurity A, B, C stock: respectively weighing each impurity reference substance, adding absolute ethyl alcohol for dissolving, and diluting with the diluent with the same volume;
test solution: adding diluent into the product, adding alkaline reagent with the same volume, shaking, standing for layering, adding the diluent layer into a centrifuge tube, adding desiccant, shaking, standing, and collecting supernatant;
mixing solution: measuring impurity A, B, C, diluting the stock solution with the sample solution;
control solution: precisely weighing the mixed solution, diluting with a diluent, and shaking uniformly to obtain the final product;
taking mixed solution for sample injection, and examining the separation condition of a main peak and three structural isomer impurity peaks; and respectively sampling the sample solution and the control solution, and calculating the contents of three impurities in the sample.
Preferably, the alkaline reagent is selected from sodium hydroxide solution or potassium hydroxide solution.
Preferably, the drying agent is selected from anhydrous sodium sulfate or anhydrous magnesium sulfate.
Preferably, the diluent is dichloromethane or chloroform or a combination of both.
More preferably, the sample preparation and detection method comprises the steps of:
1mol/L sodium hydroxide solution: weighing 4.0g of sodium hydroxide, adding 100ml of water for dissolution, and shaking uniformly to obtain the product;
impurity A, B, C stock: weighing about 7.5mg of each impurity reference substance, placing into a same 100ml measuring flask, adding 50ml of absolute ethyl alcohol for dissolution, diluting to scale with a diluent, and shaking uniformly to obtain the final product;
test solution: about 50mg of the product is taken, precisely weighed and placed in a 60ml separating funnel, firstly, 10ml of diluent is precisely added, then 10ml of 1mol/L sodium hydroxide solution is precisely added, shaking is carried out to dissolve, standing is carried out for 10min to separate the diluent layer into 10ml of centrifuge tubes, then 2.0g of anhydrous sodium sulfate is added, shaking is carried out, standing is carried out, and supernatant fluid is taken to obtain the product;
mixing solution: weighing 0.5ml of impurity A, B, C stock solution, placing into a 5ml measuring flask, diluting with sample solution to scale, and shaking;
taking mixed solution for sample injection, and examining the separation condition of a main peak and three structural isomer impurity peaks; respectively sampling the sample solution and the control solution, and calculating the contents of three impurities in the sample;
the quantitative calculation formulas of the three impurities are as follows:
wherein:
A impurity(s) : corresponding impurity A, B, C peak area in the sample solution;
A for a pair of : main peak area in control solution.
Preferably, the column initiation temperature is 60 ℃, and the temperature is raised to 250 ℃ at a rate of 10 ℃ per minute for 5 minutes.
Preferably, the column flow rate is 1.0ml/min.
Preferably, the detection temperature used is 300 ℃; the temperature of the sample inlet is 250 ℃; the split ratio was 10:1.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a gas chromatography detection scheme for analyzing isomer impurities of diethyl indene amine hydrochloride, which solves the problem that the diethyl indene amine hydrochloride and three isomer impurities are not well separated in the existing method, and the diluent and the main component have no interference to the content inspection of each impurity, and the detection accuracy and precision are high; the retention time of the main peak is proper, and the impurity detection efficiency is high; the invention also researches the conditions of diluent replacement, initial temperature adjustment, temperature rising rate and impurity separation under column flow conditions in a certain range, and formulates proper chromatographic conditions.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention.
FIG. 1 is a chromatogram showing the detection and separation effect of a related substance of diethyl indene amine hydrochloride by adopting an HPLC method;
FIG. 2 is a chromatogram showing the detection and separation effect of diethyl indene amine hydrochloride provided in example 1 of the present invention by gas chromatography;
FIG. 3 is a chromatogram of the separation effect under the chromatographic conditions provided in comparative example 1;
FIG. 4 is a chromatogram of the separation effect under the chromatographic conditions provided in comparative example 2.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present invention. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.
Example 1 detection of a mixture of diethylaniline hydrochloride with impurity A, B, C
Chromatographic column: a capillary column with 100% dimethyl polysiloxane as a fixing liquid; column temperature: the initial temperature is 60 ℃, the temperature is raised to 250 ℃ at the rate of 10 ℃ per minute, and the temperature is maintained for 5 minutes;
column flow rate: 1.0ml/min;
sample injection volume: 1 μl; a detector: a hydrogen flame ionization detector (FID detector);
detecting the temperature: 300 ℃; sample inlet temperature: 250 ℃; split ratio: 10:1.
A diluent: dichloromethane;
sample preparation:
1mol/L sodium hydroxide solution: weighing 4.0g of sodium hydroxide, adding 100ml of water for dissolution, and shaking uniformly to obtain the product.
Impurity A, B, C stock: weighing about 7.5mg of each impurity reference substance, placing into a same 100ml measuring flask, adding 50ml of absolute ethyl alcohol for dissolution, diluting to scale with dichloromethane, and shaking uniformly to obtain the final product.
Test solution: about 50mg of the product is taken, precisely weighed and placed in a 60ml separating funnel, 10ml of dichloromethane is precisely added, then 10ml of 1mol/L sodium hydroxide solution is precisely added, shaking is carried out to dissolve, standing is carried out for 10min to separate layers, a dichloromethane layer is taken into a 10ml centrifuge tube, 2.0g of anhydrous sodium sulfate is added, shaking is carried out, standing is carried out, and supernatant fluid is taken to obtain the product.
Mixing solution: accurately measuring impurity A, B, C, storing 0.5ml, placing into 5ml measuring flask, diluting with sample solution to scale, and shaking.
Control solution: precisely measuring 1ml of the mixed solution, placing the mixed solution into a 100ml measuring flask, diluting to a scale with a diluent, and shaking uniformly; precisely measuring 3ml, placing into a 20ml measuring flask, diluting with diluent to scale, and shaking.
Taking the mixed solution for sample injection, and examining the separation condition of the main peak and the three structural isomer impurity peaks. As shown in the results of FIG. 2 and Table 1, the main peak and the impurity peaks of the three structural isomers can be completely separated, the peak shapes of the main peak and the impurity peak are good, the base line is stable, and the accurate quantification can be realized.
The quantitative calculation formulas of the three impurities are as follows:
wherein:
A impurity(s) : corresponding impurity A, B, C peak area in the sample solution;
A for a pair of : main peak area in control solution;
table 1 peak results for impurities in the mixed solution in example 1:
example 2 durability test
Fine tuning chromatographic conditions, taking mixed solution sample injection under the conditions of initial temperature 54 ℃, 66 ℃, heating rate 8 ℃/min, 12 ℃/min, column flow rate 0.8ml/min, 1.0ml/min (example 1) and 1.4ml/min, and examining the separation condition of main peak and three structural isomer impurity peaks, wherein the results are shown in Table 2.
TABLE 2 durability test results summary table
From the results, the minimum separation degree between the main peak and the three structural isomer impurity peaks under each condition is 1.05, and the complete separation can be achieved.
Example 3 accuracy and precision test
Accuracy and precision were tested with reference to the chromatographic conditions provided in example 1.
Sample preparation:
impurity control solution: precisely measuring 1ml of the impurity A, B, C stock solution, placing into a 10ml measuring flask, diluting to scale with dichloromethane, and shaking to obtain the final product.
Recovery of sample solution: 6 parts of the mixed solution is prepared by the same method.
Taking each solution for sample injection, checking the signal to noise ratio of the impurity peaks of the three structural isomers in the impurity reference substance solution, and calculating the recovery rate and the precision of the impurity peaks of the three structural isomers, wherein the results are shown in Table 2.
TABLE 2 summary of accuracy and precision test results
Example 4 diluent selection test
And the diluent is replaced by chloroform, and the impurity reference substance solution and the recovery rate sample solution are prepared by the same method, so that accuracy and precision tests are carried out. Taking each solution for sample injection, checking the signal to noise ratio of the impurity peaks of the three structural isomers in the impurity reference substance solution, and calculating the recovery rate and the precision of the impurity peaks of the three structural isomers, wherein the result is shown in Table 3.
TABLE 3 accuracy and precision of diluent replacement test results summary table
The result shows that under the condition of two diluents, the signal to noise ratio of the impurity peaks of the three structural isomers is more than 30, and the method meets the requirements, and shows that the detection sensitivity is high by adopting dichloromethane or chloroform or a mixture of the dichloromethane and the chloroform as the diluent; the recovery rate of the impurity peaks of the three structural isomers is 80% -120%, and meets the requirements, which shows that the accuracy of the result of detecting the impurity content of each structural isomer by the diluent provided by the method is high, and the product quality can be strictly controlled; RSD of the recovery rate of the impurities of the three structural isomers is less than 2%, which indicates that the diluent provided by the method has good precision in detecting the impurity content of each structural isomer.
In comparative example 1,
the following chromatographic conditions were used:
chromatographic column: a capillary column with 100% dimethyl polysiloxane as a fixing liquid;
column temperature: the initial temperature is 60 ℃, the temperature is raised to 250 ℃ at a rate of 20 ℃ per minute, and the temperature is maintained for 5 minutes;
column flow rate: 1.0ml/min
Sample injection volume: 1 μl;
a detector: a hydrogen flame ionization detector (FID detector); detecting the temperature: 300 ℃;
sample inlet temperature: 250 ℃;
split ratio: 10:1.
A diluent: dichloromethane.
Taking the mixed solution of the example 1 for sample injection, and examining the separation condition of main peaks and three structural isomer impurity peaks. As a result, as shown in fig. 3, the temperature rise rate was too high, resulting in impurity A, C, and incomplete separation between impurity a and the main peak.
Comparative example 2
Chromatographic conditions: chromatographic conditions as in example 1
The sample was prepared directly using a different sample preparation method than example 1, without sodium hydroxide treatment:
impurity A, B, C stock: weighing about 7.5mg of each impurity reference substance, placing into a same 100ml measuring flask, adding 50ml of absolute ethyl alcohol for dissolution, diluting to scale with dichloromethane, and shaking uniformly to obtain the final product.
Mixing solution: accurately measuring 1ml of impurity A, B, C stock solution, placing into a 10ml measuring flask with 50mg of sample, adding absolute ethanol for dissolving, diluting to scale with dichloromethane, and shaking.
And (5) sampling the prepared mixed solution, and examining the separation condition of the main peak and the impurity peaks of the three structural isomers. As a result, as shown in fig. 4, the impurity a and the main peak cannot be completely separated, and deterioration of the sensitivity of the impurity peak occurs. The analytical reasons may be that the product prepared by the method exists in the form of hydrochloride, the boiling point is higher, the peak is generated slowly, the peak shape is wide, the separation degree of chromatographic peaks is influenced, and the sensitivity corresponding to the reduction of the peak height is poor.
When the analysis method provided by the invention is used for preparing a sample, alkaline reagents such as sodium hydroxide, potassium hydroxide, ammonium hydroxide, ammonia water, potassium carbonate, potassium bicarbonate and the like are added to react with hydrochloric acid to generate diethyl indene amine free base, so that on one hand, the boiling point of a sample to be tested can be reduced, the detection of components is facilitated, and meanwhile, the effect of increasing the solubility of substances in a diluent can be achieved.
Because weak base reaction is slow and incomplete, carbonator can generate bubbles, which affect product preparation.
Anhydrous sodium sulfate is a desiccant, and may be replaced with anhydrous magnesium sulfate.
The invention adopts GC method to realize complete separation between main component and three structural isomer impurity chromatographic peaks, and the diluent and main component have no interference to the inspection of the content of each impurity. The method has the advantages of strong specificity, high accuracy and sensitivity, good precision and durability, simple sample preparation and analysis operation and high detection efficiency.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. An analysis method of isomer impurities in diethyl indene amine hydrochloride is characterized in that gas chromatography is adopted for detection, and a chromatographic column adopted is a capillary column;
the gas chromatography detection conditions were as follows:
column temperature: the initial temperature is 54-66 ℃, the temperature is raised to 230-270 ℃ at the rate of 8-12 ℃ per minute, and the temperature is maintained for at least 5 minutes;
column flow rate: 0.8-1.4 ml/min;
sample injection volume: 1 μl; a detector: a hydrogen flame ionization detector;
detecting the temperature: 280-300 ℃; sample inlet temperature: 230-270 ℃; split ratio: 5:1-10:1;
the capillary column takes 100% dimethyl polysiloxane or 5% phenyl-95% methyl polysiloxane as a fixing solution;
when the separation degree of each component is more than 1.05, the retention time of the main component is more than 11.3min;
the isomer impurities in the diethyl indenamine hydrochloride are impurities with the following structural formula:
。
2. the method for analyzing isomer impurities in diethyl indenamine hydrochloride according to claim 1, wherein the sample preparation and detection method comprises the following steps:
impurity A, B, C stock: respectively weighing each impurity reference substance, adding absolute ethyl alcohol for dissolving, and diluting with the diluent with the same volume;
test solution: adding diluent into the product, adding alkaline reagent with the same volume, shaking, standing for layering, adding the diluent layer into a centrifuge tube, adding desiccant, shaking, standing, and collecting supernatant;
mixing solution: measuring impurity A, B, C, diluting the stock solution with the sample solution;
control solution: precisely weighing the mixed solution, diluting with a diluent, and shaking uniformly to obtain the final product;
taking mixed solution for sample injection, and examining the separation condition of a main peak and three structural isomer impurity peaks; and respectively sampling the sample solution and the control solution, and calculating the contents of three impurities in the sample.
3. The method for analyzing isomer impurities in diethyl indenamine hydrochloride according to claim 2, wherein the alkaline reagent is selected from sodium hydroxide solution or potassium hydroxide solution; the drying agent is selected from anhydrous sodium sulfate or anhydrous magnesium sulfate.
4. The method for analyzing isomer impurities in diethyl indenamine hydrochloride according to claim 2, wherein the diluent is methylene chloride or chloroform or a combination of both.
5. The method for analyzing isomer impurities in diethyl indenamine hydrochloride according to claim 1, wherein the sample preparation method comprises the following steps:
1mol/L sodium hydroxide solution: weighing 4.0g of sodium hydroxide, adding 100ml of water for dissolution, and shaking uniformly to obtain the product;
impurity A, B, C stock: weighing about 7.5mg of each impurity reference substance, placing into a same 100ml measuring flask, adding 50ml of absolute ethyl alcohol for dissolution, diluting to scale with a diluent, and shaking uniformly to obtain the final product;
test solution: taking 50mg of the product, precisely weighing, placing in a 60ml separating funnel, precisely adding 10ml of diluent, precisely adding 10ml of 1mol/L sodium hydroxide solution, shaking for dissolving, standing for 10min for layering, taking a diluent layer into a 10ml centrifuge tube, adding 2.0g of anhydrous sodium sulfate, shaking, standing, and taking supernatant to obtain the product;
mixing solution: weighing 0.5ml of impurity A, B, C stock solution, placing into a 5ml measuring flask, diluting with sample solution to scale, and shaking;
taking mixed solution for sample injection, and examining the separation condition of a main peak and three structural isomer impurity peaks; respectively sampling the sample solution and the control solution, and calculating the contents of three impurities in the sample;
the quantitative calculation formulas of the three impurities are as follows:
wherein:
A impurity(s) : corresponding impurity A, B, C peak area in the sample solution;
A for a pair of : main peak area in control solution.
6. The method for analyzing isomer impurities in diethyl indenamine hydrochloride according to claim 1, wherein the chromatographic column is started at 60 ℃, and is heated to 250 ℃ at a rate of 10 ℃ per minute for 5 minutes.
7. The method for analyzing isomer impurities in diethyl indenamine hydrochloride according to claim 1, wherein the flow rate of the chromatographic column is 1.0ml/min.
8. The method for analyzing isomer impurities in diethyl indenamine hydrochloride according to claim 1, wherein the detection temperature is 300 ℃; the temperature of the sample inlet is 250 ℃; the split ratio was 10:1.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR1456957A (en) * | 1964-09-28 | 1966-07-08 | Givaudan & Cie Sa | Process for the preparation of indane derivatives |
| US5155272A (en) * | 1976-12-29 | 1992-10-13 | Monsanto Company | Process for the production of haloacylamides |
| CN107735390A (en) * | 2015-01-20 | 2018-02-23 | 欧伦股份公司 | The preparation method of indane amine derivative and new synthetic intermediate |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR1456957A (en) * | 1964-09-28 | 1966-07-08 | Givaudan & Cie Sa | Process for the preparation of indane derivatives |
| US5155272A (en) * | 1976-12-29 | 1992-10-13 | Monsanto Company | Process for the production of haloacylamides |
| CN107735390A (en) * | 2015-01-20 | 2018-02-23 | 欧伦股份公司 | The preparation method of indane amine derivative and new synthetic intermediate |
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| Title |
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| ASHVIN D. PANCHAL et al..Synthesis of N-(5-(Substitutedphenyl)- 4,5-dihydro-1H-pyrazol-3-yl)-4H-1,2,4-triazol- 4-amine from 4-Amino-4H-1,2,4-triazole.《E-Journal of Chemistry》.2011,第8卷(第3期),全文. * |
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