CN106338564B - A method of for detecting enantiomter in vildagliptin intermediate - Google Patents
A method of for detecting enantiomter in vildagliptin intermediate Download PDFInfo
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- CN106338564B CN106338564B CN201610856455.2A CN201610856455A CN106338564B CN 106338564 B CN106338564 B CN 106338564B CN 201610856455 A CN201610856455 A CN 201610856455A CN 106338564 B CN106338564 B CN 106338564B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Abstract
A kind of method for detecting enantiomter in vildagliptin intermediate of the invention, the chromatographic condition used for：Chromatographic column is CHIRALPAK IC chromatographic columns, and column temperature is 25~40 DEG C, Detection wavelength 210nm, and mobile phase is the mixture of n-butanol, ethyl alcohol and diethylamine, and flow velocity is 0.8~1.2mL/min.The quantitative analysis of enantiomter (2R) 1 (2 chloracetyl) 2 Pyrrolidinecarbonitiles in (2S) 1 (2 chloracetyl) 2 Pyrrolidinecarbonitiles can be accurately carried out using the method for the present invention; the present invention effectively can carry out separation detection to enantiomter; peak type is symmetrical; without trailing phenomenon; separating degree, specificity, quantitative limit are with detection limit, linear, precision, accuracy, stability of solution, durability etc. through verification in detail; and every verification result meets the requirement of relevant laws and regulations and guideline, detection result is good.
Description
Technical field
The invention belongs to pharmaceutical technology fields, are related to a kind of side for detecting enantiomter in vildagliptin intermediate
Method, and in particular to the efficient liquid phase detection method of enantiomter in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles.
Background technology
Vildagliptin (Vildagliptin), and entitled (the S) -1- of chemistry [2- (3- hydroxyl -1- adamantane -1- bases amino] second
Acyl group] pyrrolidines -2- formonitrile HCNs.Vildagliptin by with DPP-4 in conjunction with the activity for inhibiting the enzyme by forming DPP-4 compounds,
GLP-l concentration is improved, while promoting B cell to generate insulin, reduces Glucagon concentrations, to reduce blood glucose,
And weight is had no significant effect.The chemical structural formula of vildagliptin is：
It is rapid that healthy human body pharmacokinetic studies show that oral vildagliptin absorbs, and bioavilability is about 85%.When up to peak
Between for 1~2h after administration, plasma half-life be 1.5~4.5h, protein binding rate is low (4%~17%).Its physiological disposition has
Linear pharmacokinetic feature does not repeatedly occur drug accumulation after oral medication, and pharmacokinetic parameters are not by food effect.
By literature search, the vildagliptin synthesis reported is mainly using L- prolineamides or L-PROLINE as starting material
Two kinds of routes：(1) using L-PROLINE as starting material synthetic route：The route turns using L-PROLINE as raw material through acylation, carboxyl
Become amide groups, amide groups is dehydrated to obtain key intermediate (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles, last and 3- ammonia
Vildagliptin is obtained by the reaction in base -1- adamantanols.(2) using L- prolineamides as starting material synthetic route：The route is with L- dried meat
Glutamine is starting material, is reacted with chloracetyl chloride, and midbody acid amide compound dehydration is prepared into key intermediate (2S)-
1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles, the intermediate react that be prepared into vildagliptin thick with 3- amino-1-adamantane alcohols
Product, then through refined and etc. vildagliptin.The key intermediate of two lines is (2S) -1- (2- chloracetyls) -2- pyrroles
Alkane formonitrile HCN is coughed up, the chemical structural formula for being somebody's turn to do (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles is：
In vildagliptin final product quality standard, quality control, standard have been carried out to its enantiomter R- vildagliptins
To be no more than 0.5%.And enantiomter D- prolineamide or L- of its enantiomter in material L- prolineamides
Enantiomter D-PROLINE in proline further leads to vildagliptin key intermediate (2S) -1- (2- chlorine after reaction
Acetyl group) -2- Pyrrolidinecarbonitiles enantiomter (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles generation.To ensure
Clinical application it is safe and effective, according to chiral drug guideline, it is necessary to vildagliptin key intermediate (2S) -1- (2-
Chloracetyl) (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles in -2- Pyrrolidinecarbonitiles are effectively controlled.
Existing literature reports the effective control method of chirality of intermediate (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles
It is less, Chinese Journal of Pharmaceuticals (2012,43 (12)：It 965-967) mentions using CHIRALPAKAD-H (models：Long 250mm,
Internal diameter 4.6mm, 5 μm), mobile phase (n-hexane-isoamyl alcohol-ethyl alcohol-ammonium hydroxide=70:15:15:0.05), 25 DEG C of column temperature, flow velocity
1.2ml/min, Detection wavelength 210nm, to (2R) -1- (2- chloroethenes in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles
Acyl group) -2- Pyrrolidinecarbonitiles are detected.It is tested with the method, there are peak type symmetry is bad, hangover, column imitates difference etc.
Problem, and do not provide detailed detection method.This detection method is finely adjusted, the above problem can not be also improved.Therefore, it is
It is continuously improved safety and the validity of vildagliptin, there is an urgent need to establish a kind of easy to operate, high sensitivity, favorable reproducibility
The quantitative detection side of the enantiomter of key intermediate (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles in vildagliptin
Method.
Invention content
In consideration of it, it is an object of the invention to propose a kind of side for detecting enantiomter in vildagliptin intermediate
Method, specially a kind of (2R) -1- (2- chloracetyls that can effectively detect in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles
Base) -2- Pyrrolidinecarbonitiles new method, this detection method use high performance liquid chromatography, to (2S) -1- (2- chloracetyls) -
Enantiomter (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles are effectively separated and are determined in 2- Pyrrolidinecarbonitiles
Amount, separating degree, specificity, quantitative limit and detection limit, linear, precision, accuracy, stability of solution, durability etc.
Through verification in detail, and every verification result meets the requirement of relevant laws and regulations and guideline, actually detected to work well.
To achieve the above object, the technical proposal of the invention is realized in this way：
A method of for detecting enantiomter in vildagliptin intermediate, the vildagliptin intermediate is
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles, the enantiomter are (2R) -1- (2- chloracetyls) -2- pyrrolidines
Formonitrile HCN the described method comprises the following steps：
S1, preparation (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile test solutions：
It weighs (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles samples and mobile phase is hybridly prepared into test solution,
For use；
S2, preparation (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile reference substance solutions：
It weighs (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles and mobile phase is hybridly prepared into reference substance solution, wait for
With；
S3, mixed solution is prepared：
Weigh (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles and prepared (2R) -1- (the 2- chloracetyls of step S2
Base) -2- Pyrrolidinecarbonitile reference substance solutions be added volumetric flask in, with flowing phase dilution, be uniformly mixed, mixed solution is made, waits for
With；
Reference substance solution, test solution and the mixed solution of S4, respectively draws equal amounts, inject in high performance chromatograph and carry out
It measures, the determination condition of the high performance liquid chromatography includes：Chromatographic column is CHIRALPAK IC chromatographic columns, and column temperature is 25~40
DEG C, the flow velocity of Detection wavelength 210nm, mobile phase are 0.8~1.2mL/min；
The mobile phase is the mixture of n-butanol, ethyl alcohol and diethylamine, is calculated by volume, n-butanol：Ethyl alcohol：Diethyl
Amine=58~62：38~42：0.05.
Further, it in the step S1 test solutions, is counted according to mass volume ratio (g/L), (2S) -1- (2- chloroethenes
Acyl group) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 0.1~5：1.
Further, it in the step S1 test solutions, is counted according to mass volume ratio (g/L), (2S) -1- (2- chloroethenes
Acyl group) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 1：1.
Further, in the step S2 reference substance solutions, according to mass volume ratio meter, (2R) -1- (2- chloracetyls) -
2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 0.01~1：1.
Further, it in the step S2 reference substance solutions, is counted according to mass volume ratio (g/L), (2R) -1- (2- chloroethenes
Acyl group) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 0.1：1.
Further, the step S3 according to mass volume ratio (g/L) in mixed solution, counting, (2S) -1- (2- chloroethenes
Acyl group) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 0.1~5：1, (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles：Stream
The ratio of dynamic phase is 0.001~0.1：1.
Further, the step S3 according to mass volume ratio (g/L) in mixed solution, counting, (2S) -1- (2- chloroethenes
Acyl group) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 1：1, (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles：Mobile phase
Ratio be 0.01：1.
Further, in the determination condition of the step S4 high performance liquid chromatographies, the type of CHIRALPAK IC chromatographic columns
Number it is：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler agent, filler material grain size are 5 μm.
Further, in the determination condition of the step S4 high performance liquid chromatographies, column temperature is 30 DEG C, the flow velocity of mobile phase
For 1mL/min.
Further, it in the mobile phase, calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=60：40：0.05.
Compared with prior art, beneficial effects of the present invention are：
The present invention provides (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles and (2R) -1- (2- chloracetyls) -2-
The method that Pyrrolidinecarbonitile carries out separation detection, the present invention uses the chromatographic condition different from the prior art, from HPLC collection of illustrative plates
There is certain distance between them to find out, can separation inspection effectively be carried out to enantiomter in vildagliptin key intermediate
It surveys, and peak type is symmetrical, no trailing phenomenon.
The detection method compareed using impurity reference substance, separating degree, specificity, quantitative limit are limited with detection, are linear, is accurate
Degree, accuracy, stability of solution, durability etc. through verification in detail, and every verification result meet relevant laws and regulations and
The requirement of guideline, it is actually detected to work well.
The present invention is highly practical, during atual detection, the inspection of (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles
Limit is surveyed up to 0.14 μ g/mL, you can be higher than 0.01% in detection (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles
(2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles, it is highly practical.Detection process is simple, quick.
High performance liquid chromatography of the present invention, included in determination condition in the range of be virtual value, i.e.,：
After taking arbitrary value in each parameter area, the enantiomter in key intermediate can accurately be also detected, and can be different to mapping
Structure body is effectively separated.During atual detection, to the adjustment of parameter and avoid human error to inspection convenient for testing staff
Resulting influence is surveyed, suitable for promoting the use of.
Description of the drawings
To describe the technical solutions in the embodiments of the present invention more clearly, make required in being described below to embodiment
Attached drawing is briefly described.
Fig. 1 is the HPLC collection of illustrative plates of the test solution of the embodiment of the present invention 1；
Fig. 2 is the HPLC collection of illustrative plates of the reference substance solution of the embodiment of the present invention 1；
Fig. 3 is the HPLC collection of illustrative plates of the mixed solution of the embodiment of the present invention 1.
Specific implementation mode
In order to carry out quality control, and the safety of continuous improvement vildagliptin drug to vildagliptin key intermediate
And validity, the present invention propose a kind of method for detecting key intermediate enantiomter in vildagliptin, wherein dimension
Key intermediate is (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles in Ge Lieting, and enantiomter is (2R) -1- (2- chlorine
Acetyl group) -2- Pyrrolidinecarbonitiles, i.e., mainly to (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles and (2R) -1- (2- chlorine
Acetyl group) -2- Pyrrolidinecarbonitiles progress separation detection.
A method of for detecting enantiomter in vildagliptin intermediate, include the following steps：
S1, preparation (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile test solutions：
It weighs (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles samples and mobile phase is hybridly prepared into test solution,
For use, it is counted according to mass volume ratio (g/L), (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 0.1
~5：1, the ratio of optimization is 1：1.
S2, preparation (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile reference substance solutions：
It weighs in (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles addition volumetric flask and mobile phase mixed preparing is pairs of
According to product solution, for use, counted according to mass volume ratio (g/L), (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles：Mobile phase
Ratio is 0.01~1：1；It is 0.1 to optimize ratio：1.
The preparation of S3, mixed solution：
It weighs (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles to be added in volumetric flask, precision measures what step S2 was prepared
(2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile reference substance solutions are added in volumetric flask and are diluted to scale with mobile phase, mix
It closes, shakes up, as mixed solution, for use.It is counted according to mass volume ratio (g/L), (2S) -1- (2- chloracetyls) -2- pyrrolidines
Formonitrile HCN：The ratio of mobile phase is 0.1~5：1, optimization ratio is 1：1；It is counted according to mass volume ratio (g/L), (2R) -1- (2- chlorine
Acetyl group) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 0.001~0.1：1, optimization ratio is 0.01：1；
Reference substance solution, test solution and the mixed solution of S4, respectively draws equal amounts, inject in high performance chromatograph and carry out
It measures, the determination condition of the high performance liquid chromatography includes：
Inspection method：2015 editions two annex high performance liquid chromatographies of Chinese Pharmacopoeia；
Chromatographic column：CHIRALPAK IC (models：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler
Agent, 5 μm of filler material grain size)；
Detector：UV detectors；
Detection wavelength：210nm；
Column temperature：25~40 DEG C, preferably 30 DEG C；
Flow velocity：0.8~1.2mL/min, preferably 1mL/min；
Mobile phase：It calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=(58~62)：(38~42)：0.05, preferably
60：40：0.05.Mobile phase employed in step S1 to S3 is identical as the step S4 mobile phases used.
During atual detection, for the high performance liquid chromatography that the present invention uses, each in determination condition
It is carried out in parameter area after suitably adjusting, can accurately also detect (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles, and
Can have to (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles and (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles
The separation detection of effect.
Illustrate specific implementation mode of the invention to enumerate with 7 exemplary embodiments below, chooses identical lot number (160104
Batch) (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples be detected.
Embodiment 1
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples that lot number is 160104 are chosen, to its (2R) -1- (2-
Chloracetyl) contents of -2- Pyrrolidinecarbonitiles is detected, includes the following steps：
S1, preparation (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile test solutions：
It weighs 25mg (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples and 25mL mobile phases is added, be uniformly mixed
Afterwards, it is configured to the solution of every 1mL 1.0mg containing vildagliptin, as test solution, for use.
S2, preparation (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile reference substance solutions：
It is appropriate to weigh (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile reference substances, accurately weighed, flowing phased soln is simultaneously
Quantitative dilution is made in every 1mL containing about the solution of 100 μ g, as (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile reference substances
Solution.
The preparation of S3, mixed solution：
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile 10mg are weighed, are added in 10mL volumetric flasks, precision measures step
(2R) -1- (2- the chloracetyls) -2- Pyrrolidinecarbonitile reference substance solutions 1mL of rapid S2 is added in volumetric flask and with being flowed phase dilution
To scale, mixing shakes up, as mixed solution, for use.
S4,10 μ L injection high performance liquid chromatographs of contrast solution are taken, adjusts detection sensitivity, makes the peak of principal component chromatographic peak
The 20%~25% of a height of full scale, then accurate measurement test solution, reference substance solution, each 10 μ L of mixed solution, are noted respectively
Enter chromatography, records chromatogram.Fig. 1, Fig. 2 and Fig. 3 are respectively (2S) -1- (2- chloracetyls) -2- pyrroles in the present embodiment
Cough up alkane formonitrile HCN test sample, (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles reference substances and mixed solution high performance liquid chromatography
Figure.The determination condition of wherein high performance liquid chromatography includes：
Chromatographic column：CHIRALPAK IC (models：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler
Agent, 5 μm of filler material grain size)；
Detector：UV detectors；
Detection wavelength：210nm；
Column temperature：30℃；
Flow velocity：1mL/min；
Mobile phase：It calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=60：40：0.05.Employed in step S1 to S3
Mobile phase it is identical as the step S4 mobile phases used.
After testing it is found that (2R) -1- (2- chloracetyls in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples
Base) -2- Pyrrolidinecarbonitile contents be 0.02%.As can be seen that (2S) -1- (2- chloracetyls) -2- pyrrolidines first from collection of illustrative plates
The separating degree of nitrile and (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles is more than 1.5.
Embodiment 2
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples for choosing lot number and 1 same batch of embodiment, to it
The content of (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles is detected, and the present embodiment is differed only in embodiment 1：
The parameter of flow velocity is different in chromatographic condition, and other testing conditions are consistent with embodiment 1.In the present embodiment, high performance liquid chromatography
The determination condition of method includes：
Chromatographic column：CHIRALPAK IC (models：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler
Agent, 5 μm of filler material grain size)；
Detector：UV detectors；
Detection wavelength：210nm；
Column temperature：30℃；
Flow velocity：0.8mL/min；
Mobile phase：It calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=60：40：0.05.
After testing it is found that (2R) -1- (2- chloracetyls in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples
Base) -2- Pyrrolidinecarbonitile contents be 0.02%.
Embodiment 3
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples for choosing lot number and 1 same batch of embodiment, to it
The content of (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles is detected, and the present embodiment is differed only in embodiment 1：
The parameter of flow velocity is different in chromatographic condition, and other testing conditions are consistent with embodiment 1.In the present embodiment, high performance liquid chromatography
The determination condition of method includes：
Chromatographic column：CHIRALPAK IC (models：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler
Agent, 5 μm of filler material grain size)；
Detector：UV detectors；
Detection wavelength：210nm；
Column temperature：30℃；
Flow velocity：1.2mL/min；
Mobile phase：It calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=60：40：0.05.
After testing it is found that (2R) -1- (2- chloracetyls in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples
Base) -2- Pyrrolidinecarbonitile contents be 0.02%.
Embodiment 4
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples for choosing lot number and 1 same batch of embodiment, to it
The content of (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles is detected, and the present embodiment is differed only in embodiment 1：
The parameter of column temperature is different in chromatographic condition, and other testing conditions are consistent with embodiment 1.In the present embodiment, high performance liquid chromatography
The determination condition of method includes：
Chromatographic column：CHIRALPAK IC (models：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler
Agent, 5 μm of filler material grain size)；
Detector：UV detectors；
Detection wavelength：210nm；
Column temperature：25℃；
Flow velocity：1mL/min；
Mobile phase：It calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=60：40：0.05.
After testing it is found that (2R) -1- (2- chloracetyls in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples
Base) -2- Pyrrolidinecarbonitile contents be 0.02%.
Embodiment 5
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples for choosing lot number and 1 same batch of embodiment, to it
The content of (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles is detected, and the present embodiment is differed only in embodiment 1：
The parameter of column temperature is different in chromatographic condition, and other testing conditions are consistent with embodiment 1.In the present embodiment, high performance liquid chromatography
The determination condition of method includes：
Chromatographic column：CHIRALPAK IC (models：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler
Agent, 5 μm of filler material grain size)；
Detector：UV detectors；
Detection wavelength：210nm；
Column temperature：40℃；
Flow velocity：1mL/min；
Mobile phase：It calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=60：40：0.05.
After testing it is found that (2R) -1- (2- chloracetyls in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples
Base) -2- Pyrrolidinecarbonitile contents be 0.02%.
Embodiment 6
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples for choosing lot number and 1 same batch of embodiment, to it
The content of (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles is detected, and the present embodiment is differed only in embodiment 1：
The parameter of mobile phase ratio is different in chromatographic condition, and other testing conditions are consistent with embodiment 1.In the present embodiment, efficient liquid
The determination condition of phase chromatography includes：
Chromatographic column：CHIRALPAK IC (models：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler
Agent, 5 μm of filler material grain size)；
Detector：UV detectors；
Detection wavelength：210nm；
Column temperature：30℃；
Flow velocity：1mL/min；
Mobile phase：It calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=62：38：0.05.
After testing it is found that (2R) -1- (2- chloracetyls in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples
Base) -2- Pyrrolidinecarbonitile contents be 0.02%.
Embodiment 7
(2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples for choosing lot number and 1 same batch of embodiment, to it
The content of (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles is detected, and the present embodiment is differed only in embodiment 1：
The parameter of mobile phase ratio is different in chromatographic condition, and other testing conditions are consistent with embodiment 1.In the present embodiment, efficient liquid
The determination condition of phase chromatography includes：
Chromatographic column：CHIRALPAK IC (models：Long 250mm, internal diameter 4.6mm, cellulose surface covalent bonding silicone filler
Agent, 5 μm of filler material grain size)；
Detector：UV detectors；
Detection wavelength：210nm；
Column temperature：30℃；
Flow velocity：1mL/min；
Mobile phase：It calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=58：42：0.05.
After testing it is found that (2R) -1- (2- chloracetyls in (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile samples
Base) -2- Pyrrolidinecarbonitile contents be 0.02%.
By embodiment 1 to the testing result of embodiment 7 it is found that in vildagliptin key intermediate (2S) -1- (2- chloracetyls
Base) -2- Pyrrolidinecarbonitiles detection process in, suitably to each parameter of chromatographic condition, such as mobile phase ratio, flow velocity, column temperature
It, can be to the enantiomter in key intermediate (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles after being adjusted
(2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles are effectively separated, and its testing result is effectively, accurately.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
With within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention god.
Claims (9)
1. a kind of method for detecting enantiomter in vildagliptin intermediate, the vildagliptin intermediate is (2S)-
1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles, the enantiomter are (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles,
It is characterized in that, the described method comprises the following steps：
S1, preparation (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile test solutions：
It weighs (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles samples and mobile phase is hybridly prepared into test solution, wait for
With；
S2, preparation (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitile reference substance solutions：
It weighs (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles and mobile phase is hybridly prepared into reference substance solution, for use；
S3, mixed solution is prepared：
Weigh (2S) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles and prepared (the 2R) -1- (2- chloracetyls)-of step S2
2- Pyrrolidinecarbonitile reference substance solutions are added in volumetric flask, with flowing phase dilution, are uniformly mixed, mixed solution are made, for use；
Reference substance solution, test solution and the mixed solution of S4, respectively draws equal amounts, inject in high performance chromatograph and are surveyed
Fixed, the determination condition of the high performance liquid chromatography includes：Chromatographic column is CHIRALPAK IC chromatographic columns, and column temperature is 25~40
DEG C, the flow velocity of Detection wavelength 210nm, mobile phase are 0.8~1.2mL/min；
The mobile phase is the mixture of n-butanol, ethyl alcohol and diethylamine, is calculated by volume, n-butanol：Ethyl alcohol：Diethylamine=
58~62：38~42：0.05.
2. the method for being used to detect enantiomter in vildagliptin intermediate according to claim 1, which is characterized in that institute
It states in step S1 test solutions, according to mass volume ratio meter, unit g/L, (2S) -1- (2- chloracetyls) -2- pyrrolidines
Formonitrile HCN：The ratio of mobile phase is 0.1~5：1.
3. the method for being used to detect enantiomter in vildagliptin intermediate according to claim 2, which is characterized in that institute
It states in step S1 test solutions, according to mass volume ratio meter, unit g/L, (2S) -1- (2- chloracetyls) -2- pyrrolidines
Formonitrile HCN：The ratio of mobile phase is 1：1.
4. the method for being used to detect enantiomter in vildagliptin intermediate according to claim 1, which is characterized in that institute
It states in step S2 reference substance solutions, according to mass volume ratio meter, unit g/L, (2R) -1- (2- chloracetyls) -2- pyrrolidines
Formonitrile HCN：The ratio of mobile phase is 0.01~1：1.
5. the method for being used to detect enantiomter in vildagliptin intermediate according to claim 4, which is characterized in that institute
It states in step S2 reference substance solutions, according to mass volume ratio meter, unit g/L, (2R) -1- (2- chloracetyls) -2- pyrrolidines
Formonitrile HCN：The ratio of mobile phase is 0.1：1.
6. the method for being used to detect enantiomter in vildagliptin intermediate according to claim 1, which is characterized in that institute
It states in step S3 mixed solutions, according to mass volume ratio meter, unit g/L, (2S) -1- (2- chloracetyls) -2- pyrrolidines first
Nitrile：The ratio of mobile phase is 0.1~5：1, (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is
0.001~0.1：1.
7. the method for being used to detect enantiomter in vildagliptin intermediate according to claim 6, which is characterized in that institute
It states in step S3 mixed solutions, according to mass volume ratio meter, unit g/L, (2S) -1- (2- chloracetyls) -2- pyrrolidines first
Nitrile：The ratio of mobile phase is 1：1, (2R) -1- (2- chloracetyls) -2- Pyrrolidinecarbonitiles：The ratio of mobile phase is 0.01：1.
8. the method for being used to detect enantiomter in vildagliptin intermediate according to claim 1, which is characterized in that institute
In the determination condition for stating step S4 high performance liquid chromatographies, column temperature is 30 DEG C, and the flow velocity of mobile phase is 1mL/min.
9. the method for being used to detect enantiomter in vildagliptin intermediate according to claim 1, which is characterized in that institute
It states in mobile phase, calculates by volume, n-butanol：Ethyl alcohol：Diethylamine=60：40：0.05.
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| CN112345668B (en) * | 2020-11-10 | 2022-06-17 | 贵州天安药业股份有限公司 | High performance liquid chromatography method for separating vildagliptin intermediate and R-type isomer |
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| A validated new chiral LC method for the enantiomeric separation of vildagliptin;Ch.Sruya Naga Malleswara Rao等;《Analytical CHEMISTRY》;20090930;第8卷(第3期);371-375 * |
| 维格列汀的合成;宋伟国等;《中国医药工业杂志》;20121231;第43卷(第12期);965-967 * |
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