阅读说明：本技术 一种罗替戈汀关键中间体的高效液相拆分方法 (High performance liquid phase resolution method of rotigotine key intermediate ) 是由 宋玉红 王亚萍 胡剑 王红秀 于 2020-07-15 设计创作，主要内容包括：本发明涉及药物分析检测技术领域,尤其为一种罗替戈汀关键中间体的高效液相拆分方法,所述方法采用高效液相色谱仪,以多糖衍生物正相涂敷型手性色谱柱,以正己烷、乙醇、二乙胺和三氟乙酸的混合溶液为流动相,流动相流速为1.0～1.5mL/min；色谱柱温度为30±5℃；检测波长为225nm；进样量为5～15uL。本发明的方法能够有效的将罗替戈汀关键中间体的R构型和S构型在色谱图中彻底分离,并准确的测出了其光学异构体的含量,解决了其质量控制问题,确保了最终产品罗替戈汀光学异构体的过程控制,为工业化生产带来了巨大的社会意义,具有很强的实用性。(The invention relates to the technical field of drug analysis and detection, in particular to a high performance liquid chromatography method for rotigotine key intermediates, which adopts a high performance liquid chromatograph, a polysaccharide derivative normal phase coating type chiral chromatographic column, a mixed solution of normal hexane, ethanol, diethylamine and trifluoroacetic acid as a mobile phase, and the flow rate of the mobile phase is 1.0-1.5 mL/min; the temperature of the chromatographic column is 30 +/-5 ℃; the detection wavelength is 225 nm; the sample injection amount is 5-15 uL. The method can effectively and thoroughly separate the R configuration and the S configuration of the rotigotine key intermediate in the chromatogram, accurately measure the content of the optical isomer, solve the quality control problem, ensure the process control of the final product rotigotine optical isomer, bring great social significance to industrial production and have strong practicability.)

1. A high performance liquid phase resolution method of rotigotine key intermediate is characterized in that: a high performance liquid chromatograph is adopted, a polysaccharide derivative normal phase coating type chiral chromatographic column is adopted, a mixed solution of normal hexane, ethanol, diethylamine and trifluoroacetic acid is used as a mobile phase, and the flow rate of the mobile phase is 1.0-1.5 mL/min; the temperature of the chromatographic column is 30 +/-5 ℃; the detection wavelength is 225 nm; the sample injection amount is 5-15 uL.

2. The high performance liquid phase resolution method of rotigotine key intermediates according to claim 1, characterized in that: the volume percentage of the mobile phase is as follows: n-hexane: ethanol: diethylamine: 94% -98% of trifluoroacetic acid: 2% -6%: 0.15%: 0.05 percent.

3. The high performance liquid phase resolution method of rotigotine key intermediate according to claim 1 or 2, characterized in that: the polysaccharide derivative normal phase coating type chiral chromatographic column is a polysaccharide derivative normal phase coating type chiral chromatographic CHIRALPAKOD, and the specification of the chromatographic column is 250mm multiplied by 4.6mm and 5 mu m.

4. The high performance liquid phase resolution method of rotigotine key intermediate according to claim 1 or 2, characterized in that: the high performance liquid chromatograph is an Agilent 1260 high performance liquid chromatograph.

5. The high performance liquid phase resolution method of rotigotine key intermediates according to any one of claims 1 to 4, which is characterized in that: the flow rate of the mobile phase is 1.2 mL/min; the temperature of the chromatographic column is 30 ℃; the detection wavelength is 225 nm; the sample size was 10 uL.

6. The method for high performance liquid phase resolution of rotigotine key intermediates according to claim 5, wherein the method comprises the following steps:

s1, taking an S-configuration rotigotine key intermediate, an R-configuration rotigotine key intermediate and a rotigotine key intermediate racemate, and respectively diluting the S-configuration solution, the R-configuration solution and the rotigotine key intermediate racemate into an S-configuration solution, an R-configuration solution and a racemate solution with the concentrations of 0.4-1.2 mg/ml and 0.4-1.2 mg/ml by using mobile phases;

and S2, respectively injecting the S configuration solution, the R configuration solution and the racemate solution into a liquid chromatograph, recording a chromatogram, and analyzing the chromatogram.

Technical Field

The invention relates to the technical field of drug analysis and detection, in particular to a high-efficiency liquid phase resolution method of a rotigotine key intermediate.

Background

The chemical name (S) -5,6,7, 8-tetrahydro-6- [ propyl [2- (2-thiophene) ethyl ] Rotigotine (Rotigotine)]Amino group]-1-Naphthol hydrochloric acidSalt, developed by schwarz biosciences (schwarz biosciences) in germany, is used for adjuvant therapy of early secondary parkinson's disease and late parkinson's disease, approved by FDA in us 5 months in 2007, and is the first transdermal patch for treating parkinsonism. The chemical name of Rotigotine (Rotigotine) is ((S) -5,6,7, 8-tetrahydro-6- (propyl (2- (2-thienyl) ethyl) amino) -1-naphthol), and the molecular formula is C₁₉H₂₅NOS, the drug is chiral, and the formula of rotigotine is shown below:

according to the report of rotigotine documents, synthesizing rotigotine, namely, taking 5-methoxy tetralone and n-propylamine as starting materials, and obtaining a racemate intermediate A through reductive amination reaction; after salifying the racemate intermediate A and a chiral acid resolution reagent, splitting to obtain an intermediate B with an S configuration; and carrying out condensation reaction and demethylation reaction on the intermediate B and thiophene ethanol to prepare rotigotine.

The intermediate of rotigotine has pharmacological activity only in the S configuration, namely (S) -5-methoxy-N-propyl-1, 2,3, 4-tetrahydronaphthalen-2-amine. The (R) -5-methoxy-N-propyl-1, 2,3, 4-tetrahydronaphthalene-2-amine may exist in the synthesized rotigotine intermediate, and the enantiomer can remain in rotigotine medicine through subsequent reaction to influence the quality of the medicine. The (S) -5-methoxy-N-propyl-1, 2,3, 4-tetrahydronaphthalene-2-amine is used as a key intermediate of rotigotine, and the configuration of the intermediate plays an important role in the quality of a final product, so that the control of the content of an optical isomer in the (S) -5-methoxy-N-propyl-1, 2,3, 4-tetrahydronaphthalene-2-amine has important significance in improving the quality of rotigotine medicaments and ensuring the medication safety of patients. Because the properties of optical isomers are close, the main detection methods of chiral enantiomers at present include an optical rotation method and a high performance liquid chromatography, and because the optical rotation method has relatively low accuracy and cannot detect the content of enantiomer impurities, the development of the high performance liquid chromatography which can effectively separate the chiral enantiomer impurities and accurately detect the content of the chiral enantiomer impurities is necessary.

Disclosure of Invention

The invention aims to provide a high-efficiency liquid phase resolution method of a rotigotine key intermediate, which aims to solve the problems in the background technology.

In order to achieve the purpose, the invention provides the following technical scheme:

a high performance liquid phase resolution method of rotigotine key intermediate is characterized in that: a high performance liquid chromatograph is adopted, a polysaccharide derivative normal phase coating type chiral chromatographic column is adopted, a mixed solution of normal hexane, ethanol, diethylamine and trifluoroacetic acid is used as a mobile phase, and the flow rate of the mobile phase is 1.0-1.5 mL/min; the temperature of the chromatographic column is 30 +/-5 ℃; the detection wavelength is 225 nm; the sample injection amount is 5-15 uL.

The method for high-efficiency liquid phase resolution of the rotigotine key intermediate comprises the following steps of: n-hexane: ethanol: diethylamine: 94% -98% of trifluoroacetic acid: 2% -6%: 0.15%: 0.05 percent.

In the method for high performance liquid resolution of the rotigotine key intermediate, the polysaccharide derivative normal phase coating type chiral chromatographic column is CHIRALPAK OD, and the specification of the chromatographic column is 250mm × 4.6mm and 5 μm.

In the method for high performance liquid resolution of the rotigotine key intermediate, the high performance liquid chromatograph is an Agilent 1260 high performance liquid chromatograph.

The high-efficiency liquid phase resolution method of the rotigotine key intermediate has the flowing phase flow rate of 1.2 mL/min; the temperature of the chromatographic column is 30 ℃; the detection wavelength is 225 nm; the sample size was 10 uL.

The high performance liquid phase resolution method of the rotigotine key intermediate comprises the following steps:

s1, taking an S-configuration rotigotine key intermediate, an R-configuration rotigotine key intermediate and a rotigotine key intermediate racemate, and respectively diluting the S-configuration solution, the R-configuration solution and the rotigotine key intermediate racemate into an S-configuration solution, an R-configuration solution and a racemate solution with the concentrations of 0.4-1.2 mg/ml and 0.4-1.2 mg/ml by using mobile phases;

and S2, respectively injecting the S configuration solution, the R configuration solution and the racemate solution into a liquid chromatograph, recording a chromatogram, and analyzing the chromatogram.

Compared with the prior art, the invention has the beneficial effects that:

according to the invention, the polysaccharide derivative normal phase coating type chiral chromatographic column is used, normal hexane, ethanol, diethylamine and trifluoroacetic acid are mixed according to a certain proportion to be used as a mobile phase, the R configuration and the S configuration of the rotigotine key intermediate can be effectively and thoroughly separated in a chromatogram, the content of an optical isomer of the rotigotine key intermediate is accurately measured, the quality control problem of the rotigotine key intermediate is solved, the process control of the final product rotigotine optical isomer is ensured, great social significance is brought to industrial production, and the rotigotine key intermediate has strong practicability. Meanwhile, the mobile phase of the method is easy to obtain, and the method is simple.

Drawings

FIG. 1 is a blank chromatogram of example 1;

FIG. 2 is a chromatogram map of the localization of the key intermediate of rotigotine in S configuration in example 1;

FIG. 3 is a chromatogram map of the location of the key intermediate of rotigotine in R configuration from example 1;

FIG. 4 is a chromatogram for separating the racemic mixture of the key intermediate of rotigotine in example 1;

FIG. 5 is a chromatogram map of the localization of the key intermediate of rotigotine in S configuration in example 2;

FIG. 6 is a chromatogram map of the location of the key intermediate of rotigotine in R configuration in example 2;

fig. 7 is a chromatogram for separating a mixed racemate of a key intermediate of rotigotine in example 2.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and all other embodiments obtained by a person of ordinary skill in the art without creative efforts based on the embodiments of the present invention belong to the protection scope of the present invention.

Referring to fig. 1-7, the present invention provides a technical solution:

a high performance liquid chromatography method for resolving rotigotine key intermediate adopts a high performance liquid chromatograph, a polysaccharide derivative normal phase coating type chiral chromatographic column and a mixed solution of normal hexane, ethanol, diethylamine and trifluoroacetic acid as a mobile phase, and comprises the following specific steps:

s1; preparing an S-configuration solution: weighing 5.00mg of S-configuration rotigotine key intermediate into a 10mL volumetric flask, dissolving the S-configuration rotigotine key intermediate by using a mobile phase as a diluent, performing constant volume by using the diluent, and fully mixing and shaking up to obtain S-configuration rotigotine intermediate solution;

s2; preparing an R configuration solution: weighing 5.16mg of R-configuration rotigotine key intermediate into a 10mL volumetric flask, dissolving the R-configuration rotigotine key intermediate by using a mobile phase as a diluent, performing constant volume by using the diluent, and fully mixing and shaking up to obtain an R-configuration rotigotine intermediate solution;

s3; preparing a racemate mixture solution: weighing 10.01mg of rotigotine key intermediate racemate into a 10mL volumetric flask, dissolving by using a mobile phase as a diluent, fixing the volume by using the diluent, and fully mixing and shaking up to obtain a racemate solution;

s4; injecting the S-configuration solution into a liquid chromatograph, and recording a chromatogram; taking the R configuration solution, performing a liquid chromatograph, and recording a chromatogram; removing the cyclone mixture solution, injecting into a liquid chromatograph, and recording the chromatogram; and (6) analyzing the chromatogram.