CN108375644A - A kind of analysis method of neuromuscular blocking agent intermediate - Google Patents
A kind of analysis method of neuromuscular blocking agent intermediate Download PDFInfo
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- CN108375644A CN108375644A CN201711268853.3A CN201711268853A CN108375644A CN 108375644 A CN108375644 A CN 108375644A CN 201711268853 A CN201711268853 A CN 201711268853A CN 108375644 A CN108375644 A CN 108375644A
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Abstract
A kind of analysis method of neuromuscular blocking agent intermediate, the method includes carrying out the separation of Mivacurium Chloride intermediate using high performance liquid chromatography.The method of the present invention can efficiently separate the intermediate 15 and impurity of Mivacurium Chloride, their own content is measured simultaneously, high sensitivity, accuracy is high, with preferable separating degree and defects inspecting limit, it can be used for the process control of Mivacurium Chloride synthesis technology, be conducive to the quality control of Mivacurium Chloride finished product, the disposable purity analysis detection that Mivacurium Chloride intermediate 15 can be completed, it is easy to operate, analytic process and analysis time are enormously simplified, analysis efficiency is improved, has saved process cycle and material resources, human cost.
Description
Technical field
The present invention relates to a kind of analysis methods of neuromuscular blocking agent intermediate.
Background technology
Neuromuscular blocking agent provides skeletal muscle relaxation effect for surgical operation and trachea cannula.Mivacurium Chloride be at present most
Short-acting nondepolarizing neuromuscular retarding agent, can be used as the adjuvant drug of general anesthesia, makes skeletal muscle relaxation, in favor of tracheae
Intubation and mechanical ventilation.Without apparent accumulation under clinical dosage, promote histamine release effect smaller, invariably to intracranial pressure and intraocular pressure
Good influence, easily controllable flesh pine concentration and range, post-operative recovery are fast.Mivacurium Chloride can flexibly apply in various operations, in addition to
General patient applies also for the patient that neuromuscular illness and blood potassium increase；It is especially had little effect in pediatric operation small
The angiocarpy of youngster is considered as the replacement medicine of Scoline.The good clinical manifestation of Mivacurium Chloride, makes it be obtained in anesthesia surgery
More and more concerns.
Mivacurium Chloride, its chemical name is Mivacurium chloride, (1R, 1'R) -2,2'- ((((E) -4- alkenyls
Suberoyl base) bis- (oxygroups)) bis- (substitutions of n-propyl -3,1- two)) bis- (6,7- dimethoxy -2- methyl-1s-(3,4,5- front threes
Oxygen benzyl) -1,2,3,4- tetrahydroisoquinoline -2- quaternary ammoniums) two villaumites, molecular formula C58H80Cl2N2O14, molecular weight 1100.17, knot
Structure formula：
Yuan Yan companies are Abbott, are listed for the first time in the U.S. within 1992, character is white or off-white powder；Have draw it is wet
Property, it is easily dissolved in water or dichloromethane, it is almost insoluble in n-hexane.
Intermediate 1~5 has been used when Mivacurium Chloride Material synthesis.By the study found that the intermediate when Mivacurium Chloride contains
When having impurity, can by condensation, the series reactions such as cyclization, restore, methylate, by these impurity and their derivative impurity
It is introduced into the Mivacurium Chloride raw material finally synthesized, to influence the quality of Mivacurium Chloride related preparations product.
If only in the quality standard of Mivacurium Chloride bulk pharmaceutical chemicals using correlation analysis method for Mivacurium Chloride bulk pharmaceutical chemicals at
The detection of product impurity can not then control the content of impurity from source.
Document (European Journal of Pharmaceutical Sciences, 5 (1997) 253-266) is reported
From the synthesis step of intermediate 4 synthetic intermediate 5, and it is simply possible to use in and monitors this step reaction and measure the pure of intermediate 5
The HPLC methods of degree.Chromatographic column is 5mm silica column (Lichrosorb, E.Merck), and wavelength uses 282nm, flowing
It is mutually acetonitrile-water-phosphoric acid (91:8:1), flow velocity is 1.3~1.5ml/min.The defect of this method is, is only applicable to intermediate
5 control, and other four intermediates are may not apply to, complete process control cannot be reached to the entire synthesis of Mivacurium Chloride
Purpose.
Document (chemical reagent, 2013,35 (1), 94~96) reports the content assaying method of intermediate 4.Chromatographic column is
C18 columns (Agilent 250mm × 4.6mm, 5 μm), mobile phase are acetonitrile-water (60:40), Detection wavelength is 281nm, and flow velocity is
1.0mL/min, column temperature are controlled at 25 DEG C, and are calculated using external standard method.This method is the content assaying method of intermediate 4,
The reference substance for also needing to take intermediate 4, could calculate its result.This method is unhelpful for the impurity determination of intermediate 4 first,
Secondly it cannot achieve the purpose that efficiently and rapidly to obtain result, data are provided for process control, therefore this method can not possibly be answered
For the process control to synthesis technology.
From the point of view of document disclosed above, the method for report is only capable of to some intermediate in above-described intermediate
Analysis detection is carried out, or is only capable of carrying out process control to some reaction step therein.And there are no detection method can be with
Quality analysis detection is carried out to this 5 intermediates of Mivacurium Chloride simultaneously, also there are no detection method can be a series of to this
Synthesis technology carries out process monitoring.
The structure of 5 intermediates is very much like, process control is carried out to this synthetic route, to this in analysis method
It is an extremely challenging thing that several substances, which carry out separation,.Also, it also to carry out quality control to each intermediate to be allowed to
It complies with standard, is also badly in need of establishing corresponding analysis method.In order to more preferably reach the process control to Mivacurium Chloride synthesis technology
With the quality control of Mivacurium Chloride intermediate, it is necessary to establish reasonable, efficient analyzing detecting method and be walked to above-mentioned synthesis technology
Suddenly the purity for carrying out control and intermediate is detected analysis.
Invention content
The present invention develops a kind of analysis method in a creative way, can carry out purity to five intermediates separately or together
Detection, and can also realize the process control to a series of synthesis steps from intermediate 1 to intermediate 5.
The present invention provides a kind of separation method of Mivacurium Chloride intermediate, which is characterized in that the method includes using high
Effect liquid phase chromatogram method carries out the step of separation of Mivacurium Chloride intermediate, in the high performance liquid chromatography, Detection wavelength 250
~300nm carries out gradient elution with mobile phase A and Mobile phase B, wherein the mobile phase A is aqueous solution:Organic solvent=
100:0~50:50 (v/v), Mobile phase B are aqueous solution:Organic solvent=0:100~50:50 (v/v), the condition of gradient elution
For：
A. Mobile phase B is changed to 45%~100% from 0%~30%, and the run time in this stage is 10~30min；
B. on the basis of stage a, Mobile phase B keeps 45%~100% constant rate, and the run time in this stage is 10
~30min；
Wherein, within each stage, the percentage of Mobile phase B changed or remains unchanged with the time, shared by mobile phase A
The sum of percentage and Mobile phase B percentage are 100%.
Optionally, above-mentioned separation method：In the high performance liquid chromatography, with octadecyl silane or octyl
Silane group silica gel is stationary phase.
Optionally, above-mentioned separation method：The mobile phase A is aqueous solution:Organic solvent=95:5~55:45(v/v).
Optionally, above-mentioned separation method：The Mobile phase B is aqueous solution:Organic solvent=5:95~45:55(v/v).
Optionally, the aqueous solution in the mobile phase A and Mobile phase B be each independently selected from pure water, containing acid buffer, contain
Alkali buffer solution, containing at least one in salt buffer, aqueous solution containing ion-pairing agent.
Optionally, the aqueous solution in the mobile phase A and Mobile phase B be each independently selected from pure water, containing acid buffer, contain
At least one of salt buffer, aqueous solution containing ion-pairing agent.
Optionally, the organic solvent in the mobile phase A and Mobile phase B is each independently selected from acetonitrile, alcohols extremely
Few one kind.
Optionally, the acid containing in acid buffer is in phosphoric acid, sulfuric acid, hydrochloric acid, formic acid, acetic acid, trifluoroacetic acid
It is at least one.
Optionally, the acid containing in acid buffer is selected from least one of phosphoric acid, formic acid, acetic acid, trifluoroacetic acid.
Optionally, a concentration of 0.005%~2% (v/v) containing aqueous acid, is chosen as 0.01%~1% (v/
v)。
Optionally, the alkali in the buffer solution containing alkali is selected from ammonia, ammonium hydroxide, sodium hydroxide, potassium hydroxide, diethylamine, three second
At least one of amine.
Optionally, the salt containing in salt buffer is selected from sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, phosphoric acid
At least one of hydrogen dipotassium, ammonium acetate, ammonium formate, sodium acetate.
Optionally, the salt containing in salt buffer is selected from sodium dihydrogen phosphate, potassium dihydrogen phosphate, ammonium acetate, ammonium formate, second
At least one of sour sodium.
Optionally, a concentration of 1.0mol/L~0.005mol/L of the salt-containing solution, be chosen as 0.5mol/L~
0.01mol/L。
Optionally, the ion pair in the aqueous solution containing ion-pairing agent is selected from sodium pentanesulfonate, sodium hexanesulfonate, heptane
At least one of sodium sulfonate, perfluorooctane sulfonate, decane sulfonate.
Optionally, a concentration of 0.001mol/L~0.05mol/L of the aqueous solution containing ion-pairing agent, is chosen as
0.002mol/L~0.01mol/L.
Optionally, the pH of the aqueous solution is 2.0~10.0, is chosen as 2.0~7.0.
Optionally, the pH value of the aqueous solution is using ammonium hydroxide, phosphoric acid, sodium hydroxide, formic acid, acetic acid, potassium hydroxide, three second
One or more in amine, diethylamine, trifluoroacetic acid are adjusted.
Optionally, the alcohols is selected from least one of methanol, ethyl alcohol, isopropanol.
Optionally, above-mentioned separation method, which is characterized in that the condition of the gradient elution is：
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 100~70 | 0~30 |
| 5 | 80~60 | 20~40 |
| 10 | 70~40 | 30~60 |
| 15 | 65~20 | 35~80 |
| 20 | 55~0 | 45~100 |
| 40 | 55~0 | 45~100 |
Or
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 100~80 | 0~20 |
| 5 | 80~60 | 20~40 |
| 15 | 70~40 | 30~60 |
| 20 | 50~20 | 50~80 |
| 30 | 30~0 | 70~100 |
| 50 | 30~0 | 70~100 |
Or
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 100~75 | 0~25 |
| 10 | 80~60 | 20~40 |
| 15 | 65~40 | 35~60 |
| 25 | 50~15 | 50~85 |
| 30 | 35~0 | 65~100 |
| 45 | 35~0 | 65~100 |
Wherein, the percentage of Mobile phase B changed or remains unchanged with the time, mobile phase A percentage and flowing
The sum of phase B percentages are 100%.
Optionally, above-mentioned separation method, it is characterised in that：
The flow velocity of the mobile phase is 0.5~2.0mL/min, is chosen as 0.5~1.5mL/min, or it is chosen as 0.7~
1.3mL/min；
The Detection wavelength is 260~290nm, is chosen as 270~290nm；
Sample size is 1-100 μ L, is chosen as 5~50 μ L, or be chosen as 20 μ L.
Optionally, above-mentioned separation method, which is characterized in that the Mivacurium Chloride intermediate includes in Mivacurium Chloride
Mesosome 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5
The present invention also provides a kind of detection methods of Mivacurium Chloride intermediate, which is characterized in that the detection method includes
The step of detection of Mivacurium Chloride intermediate is carried out using above-mentioned separation method.
It is optionally, described that detection method includes the following steps：Sample to be tested is taken, test solution is made, draws test sample
Solution is injected high performance liquid chromatograph, is measured using above-mentioned separation method；
Optionally, the preparation of the test solution includes the following steps：Sample to be tested is taken, is made of dilution a concentration of
The test solution of 0.1mg/mL~5.0mg/mL；
Optionally, the dilution is selected from least one of aqueous solution, organic solvent；
Optionally, the aqueous solution is selected from pure water, is tried containing acid buffer, buffer solution containing alkali, containing salt buffer, containing ion pair
At least one of agent aqueous solution；
Optionally, the acid containing in acid buffer is in phosphoric acid, sulfuric acid, hydrochloric acid, formic acid, acetic acid, trifluoroacetic acid
It is at least one；Optionally, the acid containing in acid buffer is selected from least one of phosphoric acid, formic acid, acetic acid, trifluoroacetic acid；
Optionally, a concentration of 0.005%~2% (v/v) containing aqueous acid, is chosen as 0.01%~1% (v/v)；
Optionally, the alkali in the buffer solution containing alkali is selected from ammonia, ammonium hydroxide, sodium hydroxide, potassium hydroxide, diethylamine, three second
At least one of amine；
Optionally, the salt containing in salt buffer is selected from sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, phosphoric acid
At least one of hydrogen dipotassium, ammonium acetate, ammonium formate, sodium acetate；Optionally, a concentration of 1.0mol/L containing salt buffer
~0.001mol/L is preferably 0.5mol/L~0.005mol/L；
Optionally, the ion pair in the aqueous solution containing ion-pairing agent is selected from sodium pentanesulfonate, sodium hexanesulfonate, heptane
At least one of sodium sulfonate, perfluorooctane sulfonate, decane sulfonate；Optionally, the concentration of the aqueous solution containing ion-pairing agent
For 0.001mol/L~0.05mol/L, it is chosen as 0.002mol/L~0.01mol/L；
Optionally, the pH of the aqueous solution is 2.0~10.0, is chosen as 2.0~7.0；
Optionally, the pH value of the aqueous solution is using ammonium hydroxide, phosphoric acid, sodium hydroxide, formic acid, acetic acid, potassium hydroxide, three second
One or more kinds of adjustings in amine, diethylamine, trifluoroacetic acid.
Optionally, above-mentioned detection method, which is characterized in that the Mivacurium Chloride intermediate includes being selected from Mivacurium Chloride
Intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5
The present invention also provides a kind of method for detecting purity of Mivacurium Chloride intermediate, which is characterized in that the purity testing
Method includes the steps that the purity testing of Mivacurium Chloride intermediate is carried out using above-mentioned detection method；Optionally, the purity
Assay method is calculated using area normalization method.
Optionally, above-mentioned method for detecting purity, which is characterized in that the Mivacurium Chloride intermediate includes being selected from Micoud chlorine
Intermediate 1, intermediate 2, intermediate 3, intermediate 4, the intermediate 5 of ammonium
The present invention also provides a kind of mass analysis methods of Mivacurium Chloride, which is characterized in that separating step including impurity,
Detecting step or determination step, wherein the separating step includes being detached according to above-mentioned separation method, the detecting step
Including being detected according to above-mentioned detection method, the determination step includes the step being measured according to above-mentioned method for detecting purity
Suddenly.
The mass analysis method of Mivacurium Chloride of the present invention can be applied in Mivacurium Chloride bulk pharmaceutical chemicals or Mivacurium Chloride preparation
The analysis of impurity.
Separation, detection and the content assaying method of the Micoud chloramines intermediate of the present invention have the following advantages：Can effectively it divide
Intermediate 1-5 from Mivacurium Chloride and impurity, while their own content is measured, high sensitivity, accuracy is high, tool
There are preferable separating degree and defects inspecting limit, it can be achieved that process to a series of synthesis steps from intermediate 1 to intermediate 5
Control, can be used in the process control of Mivacurium Chloride synthesis technology, is conducive to the quality control of Mivacurium Chloride finished product, disposable to be
The purity analysis detection of Mivacurium Chloride intermediate 1-5 can be completed, it is easy to operate, analytic process and analysis time are enormously simplified,
Analysis efficiency is improved, process cycle and material resources, human cost have been saved.
Description of the drawings
Fig. 1 is the high-efficient liquid phase chromatogram of the test solution of 1 intermediate 1 of embodiment；
Fig. 2 is the high-efficient liquid phase chromatogram of the test solution of 1 intermediate 2 of embodiment；
Fig. 3 is the high-efficient liquid phase chromatogram of the test solution of 1 intermediate 3 of embodiment；
Fig. 4 is the high-efficient liquid phase chromatogram of the test solution of 1 intermediate 4 of embodiment；
Fig. 5 is the high-efficient liquid phase chromatogram of the test solution of 1 intermediate 5 of embodiment；
Fig. 6 is the high-efficient liquid phase chromatogram of the mixed positioning solution of 1 intermediate 1~5 of embodiment.
In the figures above, ordinate is response, and unit AU, abscissa is retention time, unit min.
Specific implementation mode
The specific implementation mode of the present invention is described in detail below.It should be understood that described herein specific
Embodiment is merely to illustrate and explain the present invention, and is not intended to restrict the invention.
Embodiment 1
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica column
Mobile phase：Mobile phase A：Pure water-acetonitrile (95:5)
Mobile phase B：Pure water-acetonitrile (25:75)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 100 | 0 |
| 5 | 80 | 20 |
| 10 | 70 | 30 |
| 15 | 65 | 35 |
| 20 | 55 | 45 |
| 40 | 55 | 45 |
Wavelength：275nm
Flow velocity：0.5mL/min
Diluent：0.45% acetic acid aqueous solution (adjusting PH to 4.0 with ammonia solution)-acetonitrile (40:60)
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 2.5mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 5 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By area
Normalization method calculates.
Measurement result：
Embodiment 2
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octyl silane group silica gel column
Mobile phase：Mobile phase A：1% phosphate aqueous solution (adjusting PH to 2.0 with phosphoric acid)-methanol (75:25)
Mobile phase B：1% phosphate aqueous solution (adjusting PH to 2.0 with phosphoric acid)-methanol (10:90)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 70 | 30 |
| 5 | 60 | 40 |
| 10 | 40 | 60 |
| 15 | 20 | 80 |
| 20 | 0 | 100 |
| 40 | 0 | 100 |
Wavelength：260nm
Flow velocity：1.3mL/min
Diluent：1% phosphate aqueous solution (adjusting PH to 2.0 with phosphoric acid)-methanol (75:25)
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 0.5mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 10 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 3
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica column
Mobile phase：Mobile phase A：0.5% aqueous formic acid (adjusting PH to 4.6 with ammonia solution)-methanol (90:10)
Mobile phase B：0.5% aqueous formic acid (adjusting PH to 4.6 with ammonia solution)-methanol (30:70)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 85 | 15 |
| 5 | 65 | 35 |
| 10 | 60 | 40 |
| 15 | 40 | 60 |
| 20 | 35 | 65 |
| 40 | 35 | 65 |
Wavelength：285nm
Flow velocity：0.7mL/min
Diluent：0.5% aqueous formic acid (adjusting PH to 4.6 with ammonia solution)-methanol (90:10)
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 3.0mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each 10 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By area
Normalization method calculates.
Measurement result：
Embodiment 4
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octyl silane group silica gel column
Mobile phase：Mobile phase A：0.25% acetic acid aqueous solution (adjusting PH to 5.0 with ammonia solution)-methanol (95:5)
Mobile phase B：0.25% acetic acid aqueous solution (adjusting PH to 5.0 with ammonia solution)-methanol (10:90)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 80 | 20 |
| 5 | 65 | 35 |
| 15 | 45 | 55 |
| 20 | 20 | 80 |
| 30 | 5 | 95 |
| 50 | 5 | 95 |
Wavelength：280nm
Flow velocity：1.0mL/min
Diluent：Pure water
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 1.0mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 20 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 5
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica column
Mobile phase：Mobile phase A：0.01% trifluoroacetic acid aqueous solution (adjusting PH to 3.0 with sodium hydroxide)-methanol-acetonitrile
(60:30:10)
Mobile phase B：0.01% trifluoroacetic acid aqueous solution (adjusting PH to 3.0 with sodium hydroxide)-methanol-acetonitrile (20:60:
20)
Gradient elution：
Wavelength：269nm
Flow velocity：1.5mL/min
Diluent：0.01% trifluoroacetic acid aqueous solution (adjusting PH to 3.0 with sodium hydroxide)-methanol-acetonitrile (60:30:
10)
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 0.1mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 40 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 6
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octyl silane group silica gel column
Mobile phase：Mobile phase A：0.005mol/L biphosphates sodium water solution (adjusting PH to 7.0 with sodium hydroxide)-acetonitrile
(85:15)
Mobile phase B：0.005mol/L biphosphates sodium water solution (adjusting PH to 7.0 with sodium hydroxide)-acetonitrile (5:95)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 100 | 0 |
| 5 | 80 | 20 |
| 15 | 70 | 30 |
| 20 | 50 | 50 |
| 30 | 30 | 70 |
| 50 | 30 | 70 |
Wavelength：265nm
Flow velocity：0.6mL/min
Diluent：0.005mol/L biphosphates sodium water solution (adjusting PH to 7.0 with sodium hydroxide)-acetonitrile (85:15)
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 2.5mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 25 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 7
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica column
Mobile phase：Mobile phase A：0.01mol/L potassium dihydrogen phosphate aqueous solutions (adjusting PH to 2.5 with phosphoric acid)-methanol (90:
10)
Mobile phase B：0.01mol/L potassium dihydrogen phosphate aqueous solutions (adjusting PH to 2.5 with phosphoric acid)-acetonitrile (30:70)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 100 | 0 |
| 10 | 80 | 20 |
| 15 | 65 | 35 |
| 25 | 50 | 50 |
| 30 | 35 | 65 |
| 45 | 35 | 65 |
Wavelength：280nm
Flow velocity：1.2mL/min
Diluent：0.01mol/L potassium dihydrogen phosphate aqueous solutions (adjusting PH to 2.5 with phosphoric acid)-acetonitrile (30:70)
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 5.0mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 15 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 8
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octyl silane group silica gel column
Mobile phase：Mobile phase A：0.1mol/L formic acid aqueous ammonium (adjusting PH to 5.0 with formic acid or ammonia solution)-acetonitrile
(55:45)
Mobile phase B：0.1mol/L formic acid aqueous ammonium (adjusting PH to 5.0 with formic acid or ammonia solution)-methanol (40:60)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 75 | 25 |
| 10 | 60 | 40 |
| 15 | 40 | 60 |
| 25 | 15 | 85 |
| 30 | 0 | 100 |
| 45 | 0 | 100 |
Wavelength：285nm
Flow velocity：0.7mL/min
Diluent：0.1mol/L formic acid aqueous ammonium (adjusting PH to 5.0 with formic acid or ammonia solution)-methanol (40:60)
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 3.0mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 30 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 9
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica column
Mobile phase：Mobile phase A：1.0mol/L ammonium acetate solutions (adjusting PH to 3.0 with acetic acid)-methanol-acetonitrile (75:
20:5)
Mobile phase B：1.0mol/L ammonium acetate solutions (adjusting PH to 3.0 with acetic acid)-methanol-acetonitrile (20:40:40)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 97 | 3 |
| 10 | 80 | 20 |
| 15 | 45 | 55 |
| 25 | 20 | 80 |
| 30 | 5 | 95 |
| 45 | 5 | 95 |
Wavelength：290nm
Flow velocity：1.0mL/min
Diluent：1.0mol/L ammonium acetate solutions (adjust PH to 3.0) with acetic acid
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 2.0mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 10 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 10
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octyl silane group silica gel column
Mobile phase：Mobile phase A：0.5mol/L aqueous sodium acetate solutions (adjusting PH to 4.5 with ammonia solution)-methanol-acetonitrile
(60:30:10)
Mobile phase B：0.5mol/L aqueous sodium acetate solutions (adjusting PH to 4.5 with ammonia solution)-methanol-acetonitrile (45:50:5)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 90 | 10 |
| 10 | 70 | 30 |
| 15 | 52 | 48 |
| 25 | 40 | 60 |
| 30 | 25 | 75 |
| 45 | 25 | 75 |
Wavelength：270nm
Flow velocity：0.9mL/min
Diluent：0.5mol/L aqueous sodium acetate solutions (adjust PH to 4.5) with ammonia solution
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 0.1mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 50 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 11
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica column
Mobile phase：Mobile phase A：0.001mol/L pentanesulfonic acids sodium water solution-acetonitrile (80:20)
Mobile phase B：0.001mol/L pentanesulfonic acids sodium water solution-acetonitrile (45:55)
Gradient elution：
Wavelength：280nm
Flow velocity：1.0mL/min
Diluent：0.001mol/L pentanesulfonic acid sodium water solutions
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 0.5mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 20 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 12
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octyl silane group silica gel column
Mobile phase：Mobile phase A：0.05mol/L pentanesulfonic acids sodium water solution-methanol (90:10)
Mobile phase B：0.05mol/L pentanesulfonic acids sodium water solution-methanol (20:80)
Gradient elution：
Wavelength：282nm
Flow velocity：0.9mL/min
Diluent：0.05mol/L pentanesulfonic acids sodium water solution-methanol (90:10)
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 1.0mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 25 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 13
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica column
Mobile phase：Mobile phase A：0.002mol/L sodium hexanesulfonates aqueous solution-methanol (95:5)
Mobile phase B：The own pentanesulfonic acid sodium water solution-methanol (25 of 0.002mol/L:75)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 90 | 10 |
| 5 | 70 | 30 |
| 10 | 45 | 55 |
| 15 | 30 | 70 |
| 20 | 0 | 100 |
| 40 | 0 | 100 |
Wavelength：275nm
Flow velocity：1.1mL/min
Diluent：The own pentanesulfonic acid sodium water solutions of 0.002mol/L
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 0.8mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 20 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
Embodiment 14
Major experimental instrument and chromatographic condition：
Using 1260 type high performance liquid chromatographs of Agilent, it is temperature automatically controlled automatic to be equipped with G1312X binary pumps, G1329B
Injector, G1316A column ovens, G1315CD diode array detector, chromatography processing system use Agilent
Chemstation chromatographic work stations.
Chromatographic condition：
Chromatographic column：Octyl silane group silica gel column
Mobile phase：Mobile phase A：0.01mol/L decane sulfonic acids sodium water solution-acetonitrile (65:35)
Mobile phase B：0.01mol/L decane sulfonic acids sodium water solution-acetonitrile (10:90)
Gradient elution：
| Gradient (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 95 | 5 |
| 10 | 75 | 25 |
| 15 | 60 | 40 |
| 25 | 35 | 65 |
| 30 | 10 | 90 |
| 45 | 10 | 90 |
Wavelength：285nm
Flow velocity：1.0mL/min
Diluent：0.01mol/L decane sulfonic acid sodium water solutions
Test solution：The sample of intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 is weighed respectively, it is accurate
It is weighed, it is dissolved and is diluted with diluent respectively the single midbody solution containing 1.0mg is made in every 1mL；Take intermediate 1~5
It is appropriate to mix sample, it is accurately weighed, it is dissolved with diluent and dilutes the mixed positioning that the sample of mixing containing 0.01mg in every 1mL is made
Solution.
Measuring method：It is accurate respectively to measure each each 15 μ L of test solution, liquid chromatograph is injected, chromatogram is recorded.By face
Product normalization method calculates.
Measurement result：
This hair can be seen that the analysis test result of single midbody solution and mixed positioning solution from embodiment 1-14
Bright separation and detection method can make intermediate 1-5 be detached and be detected well separately or together, have preferable
Separating degree and defects inspecting limit.The method of the present invention disposably can be complete it can be seen from the test result of mixed positioning solution
It is detected at the purity analysis of Mivacurium Chloride intermediate 1-5, enormously simplifies analytic process and analysis time, improve analysis effect
Rate has saved process cycle and physics, human cost.The present invention side it can be seen from the test result of single midbody solution
Method also detected plurality of impurities, and in quantitative analysis intermediate 1-5, impurity peaks the content of the maximum impurity of area and
Total impurity content, therefore, separation of the invention, detection and content assaying method can to Mivacurium Chloride carry out more comprehensively and
Accurately analysis detection, can be used for the process control of Mivacurium Chloride synthesis technology, is conducive to the quality control of Mivacurium Chloride finished product.
The present invention described in detail above is preferably carried out mode, still, during present invention is not limited to the embodiments described above
Detail can carry out a variety of simple variants to technical scheme of the present invention within the scope of the technical concept of the present invention, this
A little simple variants all belong to the scope of protection of the present invention.
Claims (10)
1. a kind of separation method of Mivacurium Chloride intermediate, which is characterized in that the method includes using high performance liquid chromatography
The step of carrying out the separation of Mivacurium Chloride intermediate, in the high performance liquid chromatography, Detection wavelength is 250~300nm, with stream
Dynamic phase A and Mobile phase B carry out gradient elution, wherein the mobile phase A is aqueous solution:Organic solvent=100:0~50:50(v/
V), Mobile phase B is aqueous solution:Organic solvent=0:100~50:50 (v/v), the condition of gradient elution are：
A. Mobile phase B is changed to 45%~100% from 0%~30%, and the run time in this stage is 10~30min；
B. on the basis of stage a, Mobile phase B keeps 45%~100% constant rate, and the run time in this stage is 10~
30min；
Wherein, within each stage, the percentage of Mobile phase B changed or remains unchanged with the time, percentage shared by mobile phase A
Than being 100% with the sum of Mobile phase B percentage.
2. separation method according to claim 1, it is characterised in that：In the high performance liquid chromatography, with octadecyl
Bonded silica gel or octyl silane group silica gel are stationary phase；
The mobile phase A is aqueous solution:Organic solvent=95:5~55:45(v/v)；
The Mobile phase B is aqueous solution:Organic solvent=5:95~45:55(v/v)；
Optionally, the aqueous solution in the mobile phase A and Mobile phase B is each independently selected from pure water, delays containing acid buffer, containing alkali
Fliud flushing, containing salt buffer, at least one of aqueous solution containing ion-pairing agent；Optionally, in the mobile phase A and Mobile phase B
Aqueous solution be each independently selected from pure water, containing acid buffer, containing at least one in salt buffer, aqueous solution containing ion-pairing agent
Kind；
Optionally, the organic solvent in the mobile phase A and Mobile phase B is each independently selected from least one in acetonitrile, alcohols
Kind；
Optionally, it is described containing in acid buffer acid in phosphoric acid, sulfuric acid, hydrochloric acid, formic acid, acetic acid, trifluoroacetic acid at least
It is a kind of；Optionally, the acid containing in acid buffer is selected from least one of phosphoric acid, formic acid, acetic acid, trifluoroacetic acid；It is optional
Ground, a concentration of 0.005%~2% (v/v) containing aqueous acid, is chosen as 0.01%~1% (v/v)；
Optionally, the alkali in the buffer solution containing alkali is in ammonia, ammonium hydroxide, sodium hydroxide, potassium hydroxide, diethylamine, triethylamine
At least one；
Optionally, the salt containing in salt buffer is selected from sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, phosphoric acid hydrogen two
At least one of potassium, ammonium acetate, ammonium formate, sodium acetate；Optionally, the salt containing in salt buffer is selected from biphosphate
At least one of sodium, potassium dihydrogen phosphate, ammonium acetate, ammonium formate, sodium acetate；Optionally, the salt-containing solution is a concentration of
1.0mol/L~0.005mol/L is chosen as 0.5mol/L~0.01mol/L；
Optionally, the ion pair in the aqueous solution containing ion-pairing agent is selected from sodium pentanesulfonate, sodium hexanesulfonate, heptanesulfonic acid
At least one of sodium, perfluorooctane sulfonate, decane sulfonate；Optionally, the aqueous solution containing ion-pairing agent is a concentration of
0.001mol/L~0.05mol/L is chosen as 0.002mol/L~0.01mol/L；
Optionally, the pH of the aqueous solution is 2.0~10.0, is chosen as 2.0~7.0；
Optionally, the pH value of the aqueous solution using ammonium hydroxide, phosphoric acid, sodium hydroxide, formic acid, acetic acid, potassium hydroxide, triethylamine,
One or more in diethylamine, trifluoroacetic acid are adjusted；
Optionally, the alcohols is selected from least one of methanol, ethyl alcohol, isopropanol.
3. separation method according to claim 1 or 2, which is characterized in that the condition of the gradient elution is：
Or
Or
Wherein, the percentage of Mobile phase B changed or remains unchanged with the time, mobile phase A percentage and Mobile phase B institute
It is 100% to account for the sum of percentage.
4. separation method described in any one of claim 1 to 3, it is characterised in that：
The flow velocity of the mobile phase is 0.5~2.0mL/min, is chosen as 0.5~1.5mL/min, or be chosen as 0.7~1.3mL/
min；
The Detection wavelength is 260~290nm, is chosen as 270~290nm；
Sample size is 1-100 μ L, is chosen as 5~50 μ L, or be chosen as 20 μ L.
5. according to any one of Claims 1 to 4 separation method, which is characterized in that the Mivacurium Chloride intermediate includes
Intermediate 1, intermediate 2, intermediate 3, intermediate 4, intermediate 5 selected from Mivacurium Chloride
6. a kind of detection method of Mivacurium Chloride intermediate, which is characterized in that the detection method include using claim 1~
The step of any one of 5 separation methods carry out the detection of Mivacurium Chloride intermediate；
It is optionally, described that detection method includes the following steps：Sample to be tested is taken, test solution is made, draws test solution,
High performance liquid chromatograph is injected, is measured using any one of Claims 1 to 5 separation method；
Optionally, the preparation of the test solution includes the following steps：Sample to be tested is taken, is made of dilution a concentration of
The test solution of 0.1mg/mL~5.0mg/mL；
Optionally, the dilution is selected from least one of aqueous solution, organic solvent；
Optionally, the aqueous solution is selected from pure water, contains acid buffer, buffer solution containing alkali, containing salt buffer, water containing ion-pairing agent
At least one of solution；
Optionally, it is described containing in acid buffer acid in phosphoric acid, sulfuric acid, hydrochloric acid, formic acid, acetic acid, trifluoroacetic acid at least
It is a kind of；Optionally, the acid containing in acid buffer is selected from least one of phosphoric acid, formic acid, acetic acid, trifluoroacetic acid；It is optional
Ground, a concentration of 0.005%~2% (v/v) containing aqueous acid, is chosen as 0.01%~1% (v/v)；
Optionally, the alkali in the buffer solution containing alkali is in ammonia, ammonium hydroxide, sodium hydroxide, potassium hydroxide, diethylamine, triethylamine
At least one；
Optionally, the salt containing in salt buffer is selected from sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, phosphoric acid hydrogen two
At least one of potassium, ammonium acetate, ammonium formate, sodium acetate；Optionally, a concentration of 1.0mol/L containing salt buffer~
0.001mol/L is preferably 0.5mol/L~0.005mol/L；
Optionally, the ion pair in the aqueous solution containing ion-pairing agent is selected from sodium pentanesulfonate, sodium hexanesulfonate, heptanesulfonic acid
At least one of sodium, perfluorooctane sulfonate, decane sulfonate；Optionally, the aqueous solution containing ion-pairing agent is a concentration of
0.001mol/L~0.05mol/L is chosen as 0.002mol/L~0.01mol/L；
Optionally, the pH of the aqueous solution is 2.0~10.0, is chosen as 2.0~7.0；
Optionally, the pH value of the aqueous solution using ammonium hydroxide, phosphoric acid, sodium hydroxide, formic acid, acetic acid, potassium hydroxide, triethylamine,
One or more kinds of adjustings in diethylamine, trifluoroacetic acid.
7. detection method according to claim 6, which is characterized in that the Mivacurium Chloride intermediate includes being selected from Micoud chlorine
Intermediate 1, intermediate 2, intermediate 3, intermediate 4, the intermediate 5 of ammonium
8. a kind of method for detecting purity of Mivacurium Chloride intermediate, which is characterized in that the method for detecting purity includes using power
The step of profit requires the detection method described in 6 or 7 to carry out the purity testing of Mivacurium Chloride intermediate；Optionally, the purity is surveyed
Determine method to be calculated using area normalization method.
9. method for detecting purity according to claim 8, which is characterized in that the Mivacurium Chloride intermediate includes being selected from rice
Intermediate 1, intermediate 2, intermediate 3, intermediate 4, the intermediate 5 of library oronain
10. a kind of mass analysis method of Mivacurium Chloride, which is characterized in that the separating step, detecting step including impurity or survey
Determine step, wherein the separating step includes being detached according to any one of Claims 1 to 5 separation method, described
Detecting step includes being detected according to the detection method of claim 6 or 7, and the determination step includes according to claim
The step of 8 or 9 method for detecting purity are measured.
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| CN110361472A (en) * | 2019-07-29 | 2019-10-22 | 武汉嘉诺康医药技术有限公司 | A kind of HPLC detection method of Mivacurium Chloride intermediate MIV-G isomers |
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| CN110361472A (en) * | 2019-07-29 | 2019-10-22 | 武汉嘉诺康医药技术有限公司 | A kind of HPLC detection method of Mivacurium Chloride intermediate MIV-G isomers |
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