CN104316639B - A kind of LC-MS quantitatively detects the method for S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body - Google Patents
A kind of LC-MS quantitatively detects the method for S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body Download PDFInfo
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Abstract
The present invention discloses a kind of method that LC-MS quantitatively detects S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body, namely first mice plasma sample is carried out pre-service, then mice plasma sample after pre-service is analyzed through liquid phase, mass spectrometry successively, finally adopt quantified by external standard method, use calculated by peak area to go out S-configuration razaxaban and R-configuration razaxaban drug concentration in blood plasma.The method shortens the time of chiral separation, and analysis time is only 13min.The detection of the method is limited to 3ng/mL, quantitatively be limited to 10ng/mL, good in 0.01-10 μ g/mL scope internal linear relation, the recovery of method is between 86-112%, in a few days relative standard deviation is 5.50-8.86%, in the daytime relative standard deviation is 8.39-17.6%, meets the requirement that in Mice Body, S-configuration razaxaban and R-configuration razaxaban detection of drug concentration are analyzed.
Description
Technical field
The invention belongs to medical detection technique field, relate to a kind of method detecting razaxaban medicine S-configuration and isomeride R-structure contents thereof in blood plasma, be specifically related to the method that LC-MS quantitatively detects S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body.
Background technology
Razaxaban is a kind of new oral anticoagulation thing, it is a medicine that there is high selectivity and competitiveness and directly suppress the Xa factor in free state, but also Xa factor and the prothrombin activity of bonding state can be suppressed, direct effect is not had to platelet aggregation.It is high that it has bioavilability, and disease therapy spectrum is wide, and dose-effect relationship is stablized, convenient oral, the feature that bleeding risk is low.Razaxaban (rivaroxaban, BAY5927939, trade name: Xarelto) chemical by name: the chloro-N-{ of 5-[2-oxo-3-[4-(3-oxygen morpholine-4-base) phenyl] oxazolidone-5 (S)-Ji] methyl } thiophene-2-carboxylic acid amides.Its chemical structural formula is as follows:
At present, the mainly application RP-HPLC method (HPLC) that existing document is reported measures the content of Chinese patent medicine preparation razaxaban; Report application liquid chromatography mass coupling technique is had to measure razaxaban blood concentration in rat plasma, its pharmacokinetic parameter after the intravenous injection of calculating razaxaban, but the single configuration medicine of razaxaban that the common C18 post that the method is is analyzed, do not measure the situation of raceme administration, therefore do not go out razaxaban sample and whether change in Mice Body.
Therefore, set up the method for razaxaban raceme concentration in reliably sensitive detection blood plasma, especially liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantitatively detects the method for razaxaban content in blood plasma, can lay a solid foundation for the research of pharmacokinetics after razaxaban administration.
Summary of the invention
The object of the invention is the defect and the deficiency that overcome prior art, there is provided a kind of LC-MS quantitatively to detect the method for S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body, thus be that after razaxaban administration, the research of pharmacokinetics is laid a solid foundation.
Technical scheme of the present invention
A kind of LC-MS of the present invention quantitatively detects the method for S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body, specifically comprises following experimental procedure:
(1), the pre-service of mice plasma sample to be measured
Adopt the method for liquid-liquid extraction, make extractant with methylene chloride, get 50 μ L mice plasma samples,
Add 150 μ L methylene chloride, vortex concussion 10s, with 10000r/min rotating speed, centrifuging 10min, pipettes subnatant in 1.5mL centrifuge tube with liquid-transfering gun, uses N
2dry up, analyze by feed liquor matter after 50 μ L acetonitrile ultrasonic dissolutions;
(2), liquid phase process
High performance liquid chromatograph is the ACQUITYUPLC of Waters company
Liquid-phase chromatographic column: CHIRALPAKIC (250mm*4.6mm) 5 μm
Mobile phase: by volume percentage calculation, by acetonitrile: water: formic acid is the mixed solvent that the ratio of 97%:3%:0.1% forms, single channel isocratic elution;
Flow velocity: 0.9mL/min;
Column temperature: room temperature;
Sample size: 20 μ L;
Determined wavelength: all band scans
The method shunted after adopting post, liquid phase flow rate is 0.9mL/min, and entering mass spectrum flow velocity is 0.3mL/min;
(3), mass spectrometry method
Mass spectrometer is the QuattroPremierXE of Waters company, adopts high-purity liquid nitrogen through the nitrogen of gained of vaporizing as collision gas, atomization gas, assisted gas, taper hole gas, and maintains the vacuum state of system;
EI-MS process controling parameters is as follows:
ESI positive ion scan pattern is adopted to carry out multiple-reaction monitoring (hereinafter referred to as MRM), taper hole voltage 35.0V; Capillary voltage 3.28kv; Ion source temperature 115 DEG C; Desolventizing temperature 400 DEG C; Desolventizing airshed 600L/hr; Taper hole airshed 50L/hr, quota ion is to being 436.0/144.9;
(4), adopt quantified by external standard method, use calculated by peak area to go out the concentration of S-configuration razaxaban and R-configuration razaxaban medicine in blood plasma.
Above-mentioned LC-MS quantitatively detects S-configuration razaxaban and R-configuration razaxaban drug concentration method in Mice Body, and its detection is limited to 3ng/mL, is quantitatively limited to 10ng/mL, good in 10-10000ng/mL scope internal linear relation.The recovery of method is 86-112%, and in a few days relative standard deviation is 5.50-8.86%, in the daytime between relative standard deviation 8.39-17.6%, meets the requirement that in body, razaxaban drug concentration is analyzed.
Advantageous Effects of the present invention
A kind of LC-MS of the present invention quantitatively detects the method for S-configuration razaxaban and R-configuration razaxaban medicine in Mice Body, what liquid phase pillar adopted is bonding type cellulose derivative chiral post, mobile phase acetonitrile and the anti-phase system of water, the method shunted after adopting post, mass spectrum adopts electron spray ionisation source, with many reaction detection scanning (MRM), with the method process plasma sample of liquid-liquid extraction, compared with use C18 chromatographic column, razaxaban (S-configuration) and isomeride (R-configuration) concentration in vivo thereof can better be detected, break the limitation that C18 post can only detect the concentration of razaxaban (S-configuration), adopt liquid chromatography mass coupling technique in addition, the method shunted after using post, shorten the time analyzed and detect, analysis time is 13min, in last Mice Body, plasma sample adopts the pre-treating method of liquid-liquid extraction, compared with use methyl alcohol Direct precipitation protein method, reduce the detectability of analysis, the plasma sample of lower concentration also can be detected.
A kind of LC-MS of the present invention quantitatively detects the method for S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body, detection is limited to 3ng/mL, quantitatively be limited to 10ng/mL, good in 0.01-10 μ g/mL scope internal linear relation, the recovery of method is between 86-112%, in a few days relative standard deviation is 5.50-8.86%, and relative standard deviation is 8.39-17.6% in the daytime, meets the requirement that in Mice Body, S-configuration razaxaban and R-configuration razaxaban detection of drug concentration are analyzed.
Further, a kind of LC-MS of the present invention quantitatively detects the method for S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body, after analyzing mouse administration, blood concentration and the result of variations of time show, during raceme administration, R configuration may have half to change into S configuration, during single S configuration administration, sub-fraction is only had to be converted into R configuration.Therefore can be applicable to the research of pharmacokinetics after S-configuration razaxaban and razaxaban raceme oral administration.
Accompanying drawing explanation
The chromatogram of many reaction detection of Fig. 1, detectability 3.0ng/mL, ion pair 436.0/144.9;
Fig. 2, quantitative limit 10.0ng/mL, many reaction detection chromatogram of ion pair 436.0/144.9;
Fig. 3 a, mouse stomach S-configuration razaxaban medicine 10mg/kg dosed administration, mean blood plasma concentration-time curve;
Fig. 3 b, mouse stomach razaxaban raceme sample 10mg/kg dosed administration, mean blood plasma concentration-time curve;
Fig. 4, the many reaction detection chromatogram of razaxaban raceme plasma sample under the experiment condition groped.
Embodiment
Below in conjunction with specific embodiment, also the invention will be further elaborated by reference to the accompanying drawings, but do not limit the present invention.
embodiment 1the determination of detection method
1.1 experimental apparatuss and reagent
Instrument:
Liquid chromatograph is WatersACQUITYUPLC (Waters company), is furnished with BinarySolventManager, Column, PDA detecting device; Chromatogram pillar: CHIRALPAKIC(4.6mm Φ * 250cm5 μm) (Daicel (China) Investment Co., Ltd);
Mass spectrometer is QuattroPremierXE(Waters company), be furnished with electron spray ionisation ion gun ESI and atmosphere pressure chemical ion source (APCI), MasslynxV4.1 workstation, UltraSonicCleanerUSKType ultrasonic cleaner;
FA604A electronic balance (Shanghai Ke Tian Electron equipment Co., Ltd);
Its woods Bel instrument manufacturing company limited of vortex mixed instrument XW-80A(Haimen City)
TGL-16G high speed tabletop centrifuge (Anting Scientific Instrument Factory, Shanghai)
DC-12 Nitrogen evaporator (Shanghai ANPEL Scientific Instrument Co., Ltd.)
Liquid-transfering gun (BIOHTPROLINE) 20-200 μ l; 10-1000 μ l
Reagent:
S-configuration razaxaban medicine, R-configuration razaxaban medicine and razaxaban racemic drug (wherein R configuration and S configuration respectively account for 50%) are provided by Zhejiang Haixiang Pharmaceutical Co., Ltd., wherein the ee value of S configuration is the ee value of 99.82%, R configuration is 99.42%;
Reagent company limited of pure AR(traditional Chinese medicines group analyzed by ethanol (absolute ethyl alcohol)),
Ethanol (HPLC level, FisherScientic) normal heptane (entering to divide) (reagent company limited of traditional Chinese medicines group),
Trifluoracetic acid (Shanghai Chu Tai Chemical Co., Ltd.);
HPLC level isopropanol, normal hexane, methyl alcohol, acetonitrile (Merck company);
Methylene chloride, methenyl choloride, cyclohexane (AR, reagent company limited of traditional Chinese medicines group);
Watson distilled water.
S-configuration razaxaban reference substance storing solution:
Precision takes the S-configuration razaxaban medicine 1.0mg of white powder, is dissolved in 10ml volumetric flask, and dissolve with acetonitrile, constant volume, is mixed with the S-configuration razaxaban reference substance storing solution that concentration is 100.0 μ g/mL, 4 DEG C of Refrigerator stores;
It is appropriate that precision measures reference substance storing solution, becomes the S-configuration razaxaban standard solution of desired concn by dilution in acetonitrile;
The preparation of razaxaban raceme reference substance storing solution:
Accurate R-configuration razaxaban medicine and each 1.0mg of S-configuration razaxaban medicine taking white powder respectively, be placed in same 10ml volumetric flask, dissolve with acetonitrile, constant volume, be mixed with the razaxaban raceme reference substance storing solution that concentration is 100.0 μ g/mL, 4 DEG C of Refrigerator stores;
Zoopery: SD male mice, body weight 230 ~ 280g, is put in laboratory rearing 3 days after buying back, free food sanitation standard feed, room temperature remains on 25 DEG C.Fasting about 12h before experiment, freely drinks water.
Calculate by volume, by ethanol: molecular weight is the polyglycol of 400: water be the mixed solution that forms of 20:60:20 as solvent dilution S-configuration razaxaban medicine and razaxaban racemic drug (wherein R configuration and S configuration respectively account for 50%), be made into 1.0mg/mLS-configuration razaxaban respectively to drug solns and razaxaban raceme to each 10mL of drug solns.
Mice plasma process: the blood plasma getting the male mice after 50 μ L administrations is placed in 1.5mL tool plug plastic centrifuge tube, add acetonitrile 150 μ L, vortex mixed 10s, ultrasonic 5min, 10000r/min, 4 DEG C of centrifugal 10min, get supernatant, syringe filter is filtered, and obtains mice plasma sample to be measured, carries out the analysis of liquid matter by following condition;
1.2 liquid-phase condition
Liquid-phase chromatographic column: CHIRALPAKIC (250mm*4.6mm) 5 μm
Sample introduction 20 μ L;
Mobile phase: acetonitrile: water: formic acid=97%:3%:0.1% single channel isocratic elution
Flow velocity: 0.9mL/min column temperature: room temperature
Determined wavelength: all band scans
The method shunted after adopting post, liquid phase flow rate is 0.9mL/min, and entering mass spectrum flow velocity is about 0.3mL/min;
1.3 Mass Spectrometry Conditions
Mass spectrometer QuattroPremierXE adopts high-purity liquid nitrogen through the nitrogen of gained of vaporizing as collision gas, atomization gas, assisted gas, taper hole gas, and maintains the vacuum state of system.In this method, each mass spectrometry parameters is specific as follows: adopt ESI positive ion scan pattern to carry out multiple-reaction monitoring, taper hole voltage: 35.0V; Capillary voltage: 3.28kv; Ion source temperature: 115 DEG C; Desolventizing temperature: 400 DEG C; Desolventizing airshed: 600L/hr; Taper hole airshed: 50L/hr, the parameter of multiple-reaction monitoring is in table 1;
The parameter list of table 1, multiple-reaction monitoring
embodiment 2method validation is tested
Linear and the related coefficient of 2.1 methods
By razaxaban raceme (namely respectively accounting for 50% by S-configuration razaxaban and the R-configuration razaxaban to form) standard reserving solution of 100 μ g/mL, being diluted to concentration respectively with acetonitrile is 0.01 μ g/mL, 0.05 μ g/mL, 0.10 μ g/mL, 0.50 μ g/mL, the razaxaban raceme standard model solution of 1.00 μ g/mL, 5.00 μ g/mL, 10.00 μ g/mL;
Carry out the analysis of liquid matter according to " 1.2 liquid-phase condition " in embodiment 1 and " 1.3 Mass Spectrometry Conditions ", draw under multiple-reaction monitoring pattern, the ion current intensity of ion pair 436.0/144.9;
With S-configuration razaxaban sample concentration for horizontal ordinate, the peak area of measured ion current intensity is that ordinate does standard linear regression equation.The linear equation of gained S-configuration razaxaban sample is y=32506.58x-1603.50, and the range of linearity is 0.01-10 μ g/mL.Coefficient R
2=0.9963.
With R-configuration razaxaban sample concentration for horizontal ordinate, with the peak area of measured ion current intensity for ordinate does standard linear regression equation, the equation of linear regression of gained R-configuration razaxaban sample is y=31809.36x-1379.32, and the range of linearity is 0.01-10 μ g/mL, coefficient R
2=0.9964.Meet quantitative testing requirement.
The preparation of quality-control sample and the drafting of linearity curve: get each 50 μ L of blank mice plasma, totally 6 parts, concentration 0.01 μ g/mL is added respectively in 6 parts of blank mice plasma, 0.05 μ g/mL, 0.10 μ g/mL, 0.50 μ g/mL, 1.00 μ g/mL, the each 50 μ L of razaxaban raceme standard model storing solution of 5.00 μ g/mL, 10.00 μ g/mL, and then add methylene chloride 150 μ L respectively, vortex concussion 10s, with the centrifugal 10min of 10000r/min rotating speed, get general 150 μ about the L of subnatant, use N
2dry up, then use the acetonitrile ultrasonic dissolution of 50 μ L, carry out the analysis of liquid matter according to " 1.2 liquid-phase condition " in embodiment 1 and " 1.3 Mass Spectrometry Conditions ".
Respectively with the concentration of S-configuration razaxaban sample and R-configuration razaxaban sample for horizontal ordinate, with gained ion current peak area for ordinate, do equation of linear regression, gained linear equation is respectively:
The linear equation y=124011.92x-23633.92 of S-configuration razaxaban blood plasma mark-on sample, R
2=0.9974;
The linear equation y=125575.29x-23481.10 of R-configuration razaxaban blood plasma mark-on sample, R
2=0.9968, good in 0.01-10.0 μ g/mL scope internal linear, when mensuration 0.01 μ g/mL blood plasma mark-on sample, wherein S/N=11.1, meets quantitative requirement.
The recovery of 2.2 methods, accuracy and precision
The method recovery and precision, accuracy are tested, add in blank mice plasma sample different oneself know razaxaban raceme (namely respectively accounting for 50% by S-configuration razaxaban and the R-configuration razaxaban to form) sample of concentration, be respectively 0.10,5.00,10 μ g/mL levels, each concentration prepares 5 samples, get clean 1.5mL plastic centrifuge tube 15, add 100 μ L blank plasmas respectively, every 5 is one group, adds the razaxaban raceme standard solution 100 μ L of 0.1 μ g/mL in first group respectively; Second group of razaxaban raceme solution 100 μ L adding 5.0 μ g/mL; 3rd group adds 10.0 μ g/mL razaxaban raceme solution 100 μ L, then in three groups of solution, adds 300 μ L dichloromethane solutions respectively, and vortex concussion 10s, 10000r/min rotating speed centrifuging 10min, gets subnatant, N
2air-blowing is done, and with 100 μ L acetonitrile ultrasonic dissolutions, is mixed with 0.1 μ g/mL, 5.0 μ g/mL, 10.0 each 5 parts of μ g/mL razaxaban raceme blood plasma mark-on solution.Second day, the 3rd day according to the method described above, prepares 0.1 μ g/mL respectively, 5.0 μ g/mL, 10.0 each 1 part of μ g/mL razaxaban blood plasma mark-on solution.The analysis of liquid matter is carried out according to " 1.2 liquid-phase condition " in embodiment 1 and " 1.3 Mass Spectrometry Conditions ".Acquired results is as table 2.
Table 2 method repeatability and recovery experimental result
Under the low concentration of 0.1 μ g/mL, the recovery of method is the recovery of the high concentration of 86-89%, 5.0-10.0 μ g/mL is as can be seen from Table 2 95-112%, meets Mice Body inner analysis quantitative requirement.During low concentration, the recovery is on the low side, tends towards stability during high concentration, all meets the requirement of In vivo analysis.As shown in Table 2, in a few days relative standard deviation is between 5.50%-8.86%, and relative standard deviation is between 8.39%-17.6% in the daytime, meets the requirement of In vivo analysis.The accuracy of illustration method and precision better, fully verify the operability of institute's method for building up.From table 2, data find when Pitch-based sphere is lower, and the recovery of razaxaban is relatively little, relative standard deviation is relatively large, and this may be that error also can increase to some extent because addition is too little to such an extent as to data fluctuations is relatively large.
The concentration of sample under quantitative limit and detectability is analyzed according to " 1.2 liquid-phase condition " in embodiment 1 and " 1.3 Mass Spectrometry Conditions ", obtain ion flow graph respectively as depicted in figs. 1 and 2, as can be seen from Figure 1, S/N >=3, determine that the detection of analytical approach is limited to 3.0ng/mL, as can be seen from Figure 2, S/N >=10, that determines analytical approach is quantitatively limited to 10.0ng/mL.
embodiment 3the mensuration of actual sample
3.1, plasma sample preparation
During administration, dissolve the solvent ethanol of razaxaban medicine: PEG400: the volume ratio of water is the mixed solution of 20:60:20;
Take S-configuration razaxaban 10.37mg, dissolve with the above-mentioned mixed solution of 10.37mL, being mixed with concentration is that the S-configuration razaxaban sample of 1.0mg/mL is to drug solns;
Take S-configuration razaxaban 9.886mg and R-configuration razaxaban 9.991mg in addition to mix and be placed in sample bottle, with the above-mentioned mixed solution ultrasonic dissolution of 9.9mL, being mixed with concentration is that the razaxaban raceme sample of 1.0mg/mL is to drug solns.
Be that the dosage of 10.0mg carries out administration to mouse according to per kilogram mouse;
The S-configuration razaxaban sample utilizing concentration to be 1.0mg/mL carries out administration to drug solns to 6 mouse, each mouse difference 10min, 20min, 30min upon administration, 1h, 2h, 4h, 6h, 8h blood sampling, each time point gets 6 plasma samples altogether, i.e. 48 plasma samples;
The razaxaban raceme sample utilizing concentration to be 1.0mg/mL carries out administration to drug solns to 6 mouse, and each mouse is respectively upon administration respectively at 10min, 20min, 30min, 1h, 2h, 4h, 6h, 8h blood sampling, each time point gets 6 plasma samples altogether, i.e. 48 plasma samples;
Above-mentioned 96 blood samples altogether, according to " disposal routes of 1.1 plasma samples ", carry out the analysis of liquid matter after process plasma sample.
3.2, actual plasma sample tests
By 96 of above-mentioned gained plasma samples according to " disposal routes of 1.1 plasma samples ", process plasma sample, the analysis of liquid matter is carried out according to " 1.2 liquid-phase condition " in embodiment 1 and " 1.3 Mass Spectrometry Conditions ", the average rear acquired results of 6 values of each point brings the blood plasma mark-on equation in " the linear and related coefficients of 2.1 methods " into, the concentration of S-configuration razaxaban medicine and corresponding body R-configuration razaxaban medicine thereof in sampling under going out different time according to calculated by peak area.Acquired results is as table 3.
Sample concentration (n=6) is sampled under table 3 different time
3.3, the drafting of the change curve of mouse vivo medicine concentration and time after administration
According to the data of table 3, draw out mouse vivo medicine concentration and the change curve of time after the razaxaban administration of S-configuration and raceme administration respectively, specifically see shown in Fig. 3 a, Fig. 3 b, as can be seen from Fig. 3 a, Fig. 3 b, during the mono-configuration administration of independent S-, in Mice Body, S-configuration has sub-fraction and is converted into R-configuration, and during raceme administration, R configuration may have half to change into S configuration, therefore can be applicable to the research of pharmacokinetics after razaxaban list configuration and raceme oral administration.
The raceme blood plasma mark-on sample that above-mentioned concentration is 5.0 μ g/mL is analyzed according to " 1.2 liquid-phase condition " in embodiment 1 and " 1.3 Mass Spectrometry Conditions ", its result is as Fig. 4, and as can be seen from Figure 4 S-configuration razaxaban and R-configuration razaxaban can reach baseline separation.
In sum, LC-MS of the present invention quantitatively detects razaxaban drug concentration in sensitive, accurate, the reliable detection blood sample of method energy of razaxaban S-configuration and isomeride R-structure contents thereof in blood plasma.
The above is only the citing of embodiments of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the technology of the present invention principle; can also make some improvement and modification, these improve and modification also should be considered as protection scope of the present invention.
Claims (1)
1. LC-MS quantitatively detects a method for S-configuration razaxaban and R-configuration razaxaban drug concentration in Mice Body, it is characterized in that specifically comprising the steps:
(1), sample pretreatment
Adopt the method for liquid-liquid extraction, extractant is made with methylene chloride, get 50 μ L mice plasma samples, add 150 μ L methylene chloride, vortex concussion 10s, with 10000r/min rotating speed, centrifuging 10min, pipette subnatant in 1.5mL centrifuge tube with liquid-transfering gun, dry up with N2, analyze by feed liquor matter after 50 μ L acetonitrile ultrasonic dissolutions; Described plasma sample is that mouse stomach is to the blood plasma after drug solns; Described is that concentration is respectively 1.0mg/mL razaxaban raceme solution and S-configuration razaxaban solution to drug solns; Administration solution preparation solvent used is the polyglycol of 400 and the volume ratio of water by ethanol, molecular weight is the mixed solution that 20:60:20 forms;
(2), liquid phase analysis method
Instrument used, high performance liquid chromatograph is the ACQUITYUPLC of Waters company;
Liquid-phase chromatographic column: CHIRALPAKIC250mm*4.6mm5 μm;
Mobile phase: by volume percentage calculation, by acetonitrile: water: formic acid is the mixed solvent that the ratio of 97%:3%:0.1% forms, single channel isocratic elution;
Flow velocity: 0.9mL/min;
Column temperature: room temperature;
Sample size: 20 μ L;
Determined wavelength: all band scans;
The method shunted after adopting post, liquid phase flow rate is 0.9mL/min, and entering mass spectrum flow velocity is 0.3mL/min;
(3), mass spectrometric analysis method
Instrument used, mass spectrometer is the QuattroPremierXE of Waters company, adopts high-purity liquid nitrogen through the nitrogen of gained of vaporizing as collision gas, atomization gas, assisted gas, taper hole gas, and maintains the vacuum state of system;
EI-MS process controling parameters is as follows:
ESI positive ion scan pattern is adopted to carry out multiple-reaction monitoring, taper hole voltage 35.0V; Capillary voltage 3.28kv; Ion source temperature 115 DEG C; Desolventizing temperature 400 DEG C; Desolventizing airshed 600L/hr; Taper hole airshed 50L/hr, quota ion is to being 436.0/144.9;
(4), adopt quantified by external standard method, use calculated by peak area to go out S-configuration razaxaban and R-configuration razaxaban drug concentration in blood plasma.
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| CN110297054A (en) * | 2019-08-14 | 2019-10-01 | 西安和合医学检验所有限公司 | The detection method of Levetiracetam content in a kind of human serum |
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| CN103896933A (en) * | 2012-12-26 | 2014-07-02 | 上海医药工业研究院 | Intermediate of rivaroxaban and preparation method thereof |
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| CN103896933A (en) * | 2012-12-26 | 2014-07-02 | 上海医药工业研究院 | Intermediate of rivaroxaban and preparation method thereof |
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