CN106053657B - A method of using high performance liquid chromatography detection chirality phenyl ethylamine content - Google Patents
A method of using high performance liquid chromatography detection chirality phenyl ethylamine content Download PDFInfo
- Publication number
- CN106053657B CN106053657B CN201610513852.XA CN201610513852A CN106053657B CN 106053657 B CN106053657 B CN 106053657B CN 201610513852 A CN201610513852 A CN 201610513852A CN 106053657 B CN106053657 B CN 106053657B
- Authority
- CN
- China
- Prior art keywords
- phenyl ethylamine
- solution
- preparation
- liquid chromatography
- performance liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
A method of high performance liquid chromatography detection chirality phenyl ethylamine content being used applied to one of combinations of materials detection field, which includes the following steps: chromatographic condition: chromatographic column: the unmodified packed column of crown ether derivative coating, 5 μm of partial size；Detection wavelength: UV detector 210nm；Mobile phase: high chloro acid solution pH=1.0/ acetonitrile=50:50 (volume ratio)；The preparation of high chloro acid solution's pH=1.0 solution；The preparation of system suitability solution；The preparation of test solution；Measuring method；The invention, which detects left-handed phenyl ethylamine and dextrorotation phenyl ethylamine reproducibility, sensitivity, accuracy, can meet the requirements.
Description
Technical field
The present invention relates to one of Pharmaceutical Analysis fields using the side of high performance liquid chromatography detection chirality phenyl ethylamine content
Method.
Background technique
Phenyl ethylamine, also known as β-phenyl ethylamine are a kind of alkaloid and monoamine neurotransmitter.Phenyl ethylamine has a structural isomerism
Body, i.e. α-phenylethylamine, there are two stereoisomers for α-phenylethylamine: (R)-(+) -1- phenyl ethylamine and (S)-(-) -1- phenyl ethylamine.Hand
Property phenyl ethylamine be to prepare one of important intermediate of single enantiomer, it can not only be used for the chiral resolving agent of racemic modification, split
Organic acid, but also as the chiral auxiliary and synthesis material of asymmetric syntheses, so its preparation and analysis have important meaning
Justice.The method of inspection split at present about chiral phenyl ethylamine, has no relevant report, therefore, develops a kind of chiral phenyl ethylamine
The method of inspection be always new issue urgently to be resolved.
Summary of the invention
The purpose of the present invention is to provide a kind of method using high performance liquid chromatography detection chirality phenyl ethylamine content, the party
Method uses high effective liquid chromatography for measuring, has that favorable reproducibility, high sensitivity, detection is accurate, specificity is strong, the spies such as easy to operate
Point.
To achieve the above object, the present invention adopts the following technical scheme: a kind of use high performance liquid chromatography detection chirality benzene
The method of ethamine content, the described method comprises the following steps:
(1) chromatographic condition:
Chromatographic column: the unmodified packed column of crown ether derivative coating；
Detection wavelength: ultraviolet wavelength 210nm；
Mobile phase: the high chloro acid solution of pH=1.0 and the mixed solvent of acetonitrile；
(2) preparation of mobile phase, the preparation of system suitability solution, the preparation of test solution；
(3) measuring method:
It takes system suitability solution to inject liquid chromatograph, map is recorded, wherein left-handed phenyl ethylamine and dextrorotation phenyl ethylamine peak
Separating degree answer >=1.5, take test solution inject liquid chromatograph, record map, calculate separately left-handed in test solution
The peak area of phenyl ethylamine and the peak area of dextrorotation phenyl ethylamine；
The partial size of the silica gel of the crown ether derivative coating is 5 μm；The high chloro acid solution of the pH=1.0 and acetonitrile
Volume ratio is=45-55:45-55, preferred volume ratio 50:50；The flow velocity of the mobile phase are as follows: 0.3-0.5mL/min, preferably
Flow velocity be 0.4mL/min；The column temperature of the chromatographic column is 20-30 DEG C, and preferred column temperature is 25 DEG C；The preparation of the mobile phase
Method are as follows: perchloric acid is weighed in the first volumetric flask, with water constant volume, is mixed, as the high chloro acid solution of pH=1.0, it will be upper
The high chloro acid solution for stating pH=1.0 is transferred in the second volumetric flask, is settled to scale with acetonitrile, mixes, degassing；Described first
The volume of volumetric flask is 1 liter, and the volume of second volumetric flask is 2 liters；The preparation method of the system suitability solution is to claim
Mixed phenyl ethylamine reference substance 50mg is taken, with flowing phased soln and 10mL is diluted to, shakes up；The preparation method of the test solution
To weigh test sample 50mg, with flowing phased soln and it is diluted to 10mL, is shaken up；The system suitability solution and test solution
The sample volume for injecting liquid chromatograph is 0.5-5 μ L, and preferably sample volume is 1 μ L.
The detection method of the invention for being characterized by it.The basic principle of the detection method is left-handed phenyl ethylamine, dextrorotation benzene
Retention time of the ethamine in mobile phase is different, calculates its content using HPLC area normalization method.
A kind of method using high performance liquid chromatography detection chirality phenyl ethylamine content compared with prior art, has and reappears
The advantages that property, sensitivity, accuracy can meet the requirements, will be widely used in Pharmaceutical Analysis field.
Detailed description of the invention
The following describes the present invention in detail with reference to the accompanying drawings and embodiments.
Fig. 1 is the HPLC test map of test sample 1.
Fig. 2 is the HPLC test map of test sample 2.
Fig. 3 is the HPLC test map of test sample 3.
Specific embodiment
The present invention will be further explained with reference to the examples below, and embodiment helps to more fully understand the present invention, but
The present invention is not limited only to following embodiments.
Embodiment one
1. chromatographic condition:
Chromatographic column: the unmodified packed column of crown ether derivative coating, 5 μm of partial size；
Detection wavelength: ultraviolet wavelength 210nm；
Mobile phase: the high chloro acid solution of pH=1.0/acetonitrile=50:50 (volume ratio)；
Flow velocity: 0.4mL/min；
Column temperature: 25 DEG C；
2. the preparation of mobile phase, system suitability solution and test solution:
The preparation of 2.1 mobile phases: weighing 16.3g perchloric acid (content 72%) in 1000mL brown volumetric flask, fixed with water
Hold, mixes, as the high chloro acid solution of 1L pH=1.0.The high chloro acid solution of above-mentioned 1L pH=1.0 is transferred to 2L to hold
In measuring bottle, it is settled to scale with acetonitrile, is mixed, degassing.
The preparation of 2.2 system suitability solution:
Mixed phenyl ethylamine reference substance 50mg is weighed, with flowing phased soln and 10mL is diluted to, shakes up；
The preparation of 2.3 test solutions:
2.3.1 test sample 1 is weighed, (R)-(+) -1- phenyl ethylamine 50mg with flowing phased soln and is diluted to 10mL, filters.
2.3.2 test sample 2 is weighed, mixed phenyl ethylamine 50mg with flowing phased soln and is diluted to 10mL, filters.
2.3.3 test sample 3 is weighed, right phenyl ethylamine salt 50mg with flowing phased soln and is diluted to 10mL, filters.
3. measuring method:
It takes 1 μ L of system suitability solution to inject liquid chromatograph, records map, point of left-handed phenyl ethylamine and dextrorotation phenyl ethylamine
>=1.5 are answered from degree, 3 batches of test liquids, 1 μ L is respectively taken to inject liquid chromatograph, map is recorded, calculates separately left-handed benzene second in test liquid
The peak area of amine and the peak area of dextrorotation phenyl ethylamine.
Inspection result:
Test sample 1:(R)-(+) -1- phenyl ethylamine (HPLC chromatogram is shown in attached drawing 1):
(S)-(-) area of -1- phenyl ethylamine is 25091, and the area of (R)-(+) -1- phenyl ethylamine is 3373982
(R)-(+) -1- phenyl ethylamine content is [3373982 ÷ (25091+3373982)] × 100%=99.26%
(S)-(-) -1- phenyl ethylamine content is [25091 ÷ (25091+3373982)] × 100%=0.74%；
Test sample 2: mixed phenyl ethylamine (HPLC chromatogram is shown in attached drawing 2):
(S)-(-) area of -1- phenyl ethylamine is 13643358, and the area of (R)-(+) -1- phenyl ethylamine is 15269336
(R)-(+) -1- phenyl ethylamine content is [15269336 ÷ (13643358+15269336)] × 100%=52.81%
(S)-(-) -1- phenyl ethylamine content is [13643358 ÷ (13643358+15269336)] × 100%=
47.19%；
Test sample 3: right phenyl ethylamine salt (HPLC chromatogram is shown in attached drawing 3):
(S)-(-) area of -1- phenyl ethylamine is 425.3, and the area of (R)-(+) -1- phenyl ethylamine is 11903.6
(R)-(+) -1- phenyl ethylamine content is [11903.6 ÷ (425.3+11903.6)] × 100%=96.55%
(S)-(-) -1- phenyl ethylamine content is [425.3 ÷ (425.3+11903.6)] × 100%=3.45%.
Claims (5)
1. a kind of method using high performance liquid chromatography detection chirality phenyl ethylamine content, which is characterized in that the method includes with
Lower step:
(1) chromatographic condition:
Chromatographic column: the unmodified packed column of crown ether derivative coating；
Detection wavelength: ultraviolet wavelength 210nm；
Mobile phase: the high chloro acid solution of pH=1.0 and the mixed solvent of acetonitrile；
(2) preparation of mobile phase, the preparation of system suitability solution, the preparation of test solution；
(3) measuring method:
It takes system suitability solution to inject liquid chromatograph, records map, wherein point of left-handed phenyl ethylamine and dextrorotation phenyl ethylamine peak
>=1.5 are answered from degree, test solution is taken to inject liquid chromatograph, map is recorded, calculates separately left-handed benzene second in test solution
The peak area of amine and the peak area of dextrorotation phenyl ethylamine；
The flow velocity of the mobile phase are as follows: 0.3-0.5mL/min；
The partial size of the silica gel of the crown ether derivative coating is 5 μm；The high chloro acid solution of the pH=1.0 and the volume of acetonitrile
Than for=45-55:45-55；The column temperature of the chromatographic column is 20-30 DEG C；
The preparation method of the system suitability solution is to weigh mixed phenyl ethylamine reference substance 50mg, with flowing phased soln and is diluted
To 10mL, shake up；The preparation method of the test solution is to weigh test sample 50mg, with flowing phased soln and is diluted to
10mL shakes up.
2. a kind of method using high performance liquid chromatography detection chirality phenyl ethylamine content according to claim 1, feature
It is, the high chloro acid solution of the pH=1.0 and the volume ratio of acetonitrile are=50:50；The flow velocity of the mobile phase are as follows:
0.4mL/min；The column temperature of the chromatographic column is 25 DEG C.
3. a kind of method using high performance liquid chromatography detection chirality phenyl ethylamine content according to claim 1, feature
It is, the preparation method of the mobile phase are as follows: weigh perchloric acid in the first volumetric flask, with water constant volume, mix, as pH=
The high chloro acid solution of above-mentioned pH=1.0 is transferred in the second volumetric flask, is settled to acetonitrile by 1.0 high chloro acid solution
Scale mixes, degassing；The volume of first volumetric flask is 1 liter, and the volume of second volumetric flask is 2 liters.
4. a kind of method using high performance liquid chromatography detection chirality phenyl ethylamine content according to claim 1, feature
It is, the sample volume of the system suitability solution and test solution injection liquid chromatograph is 0.5-5 μ L.
5. a kind of method using high performance liquid chromatography detection chirality phenyl ethylamine content according to claim 4, feature
It is, the sample volume of the system suitability solution and test solution injection liquid chromatograph is 1 μ L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610513852.XA CN106053657B (en) | 2016-07-01 | 2016-07-01 | A method of using high performance liquid chromatography detection chirality phenyl ethylamine content |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610513852.XA CN106053657B (en) | 2016-07-01 | 2016-07-01 | A method of using high performance liquid chromatography detection chirality phenyl ethylamine content |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106053657A CN106053657A (en) | 2016-10-26 |
| CN106053657B true CN106053657B (en) | 2018-12-04 |
Family
ID=57200653
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610513852.XA Active CN106053657B (en) | 2016-07-01 | 2016-07-01 | A method of using high performance liquid chromatography detection chirality phenyl ethylamine content |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106053657B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104860830A (en) * | 2015-04-08 | 2015-08-26 | 东北制药集团股份有限公司 | Preparation method for R-(+)-alpha-phenylethylamine salt and R-(+)-alpha-phenylethylamine |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH082844B2 (en) * | 1989-06-26 | 1996-01-17 | 宇部興産株式会社 | Method for producing phenethylamines |
| WO2006126498A1 (en) * | 2005-05-23 | 2006-11-30 | Kaneka Corporation | Novel amino group transferase, gene encoding the same and method of using the same |
| US9278904B2 (en) * | 2013-12-31 | 2016-03-08 | Chemapotheca, Llc | Synthesis of chiral amphetamine derivatives by stereospecific, regioselective cuprate addition reaction with aziridine phosphoramidate compounds |
-
2016
- 2016-07-01 CN CN201610513852.XA patent/CN106053657B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104860830A (en) * | 2015-04-08 | 2015-08-26 | 东北制药集团股份有限公司 | Preparation method for R-(+)-alpha-phenylethylamine salt and R-(+)-alpha-phenylethylamine |
Non-Patent Citations (2)
| Title |
|---|
| α-苯乙胺的间接高效液相色谱拆分研究;唐琴 等;《湖北民族学院学报（自然科学版）》;20060630;第24卷(第2期);全文 * |
| 高效液相色谱手性冠醚固定相拆分丙氨酸、正缬氨酸和犬尿氨酸对映体;李齐 等;《分析化学研究简报》;20090630;第37卷;全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106053657A (en) | 2016-10-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Preinerstorfer et al. | Advances in enantioselective separations using electromigration capillary techniques | |
| Sun et al. | Ionic liquids in analytical chemistry | |
| Choi et al. | Single-drop microextraction in bioanalysis | |
| ALOthman et al. | Single-drop microextraction as a powerful pretreatment tool for capillary electrophoresis: A review | |
| CN104634887B (en) | A method of separation and measurement ticagrelor and its optical isomer | |
| CN104792913B (en) | A kind of Apremilast and the method for separating and detecting of enantiomer thereof | |
| CN110849980A (en) | Method for detecting content of enantiomer in isopropyl L-alanine | |
| CN106770855A (en) | A kind of method for separating and detecting of Sha Kubi song intermediates and its optical isomer based on high performance liquid chromatography | |
| CN110687229A (en) | Diclofenac sodium raw material and analysis method of related substances in preparation thereof | |
| Carrasco-Correa et al. | Evaluation of 2, 3-epoxypropyl groups and functionalization yield in glycidyl methacrylate monoliths using gas chromatography | |
| CN105806963B (en) | The method for separating and detecting of L dried meat ammonia alcohol and its enantiomter | |
| CN106338564B (en) | A method of for detecting enantiomter in vildagliptin intermediate | |
| CN106053657B (en) | A method of using high performance liquid chromatography detection chirality phenyl ethylamine content | |
| CN104833740A (en) | HPLC (High Performance Liquid Chromatography) method for rivaroxaban intermediate | |
| CN101929988B (en) | Method for detecting febuxostat-associated matters by using high performance liquid chromatography | |
| CN104458945B (en) | The method of separating and assaying of a kind of besifloxacin hydrochloride and isomeride thereof | |
| CN108956827A (en) | A kind of method HPLC method analysis and prepare 3- (N- p-toluenesulfonyl-L- alanyl oxygroup) indoles and its enantiomter | |
| CN105572240A (en) | Method for detecting content of pharmaceutic adjuvant carmine by using high performance liquid chromatography | |
| CN110031583B (en) | Liquid chromatography method for separating and measuring N-succinyl tryptophan enantiomer | |
| CN100535657C (en) | Method for measuring purity of 9-fluorenemethanol | |
| CN105628841A (en) | Method for separating and measuring Apremilast and enantiomer of Apremilast through liquid chromatography | |
| CN110412188A (en) | A method of the isomer impurities of separation and measurement ticagrelor intermediate | |
| Tiritan et al. | Pirkle Type | |
| CN104730160B (en) | A kind of Acetyl Tyrosine and the high-efficiency liquid chromatography method for detecting of optical isomer thereof | |
| Cheong et al. | Open tubular molecular imprinted phases in chiral capillary electrochromatography |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |