CN105136957A - Detection method for simultaneously measuring OXC in human plasma and metabolite MHD and MHD-G - Google Patents
Detection method for simultaneously measuring OXC in human plasma and metabolite MHD and MHD-G Download PDFInfo
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Abstract
The invention relates to a detection method for simultaneously measuring OXC in human plasma and metabolite MHD and MHD-G, and belongs to the technical field of biological detection. An experiment is carried out by taking nitrazepam as an internal standard, and can simultaneously detect the plasma concentrations of the three compounds by adopting high performance liquid chromatography-tandem mass spectrometry after pretreatment through precipitation protein. The detection method is short in time, high in specificity and sensitivity, and convenient in operation, and is successfully applied to the analysis and research of oxcarbazepine in the human plasma and the metabolite thereof.
Description
Technical field
The present invention relates to the detection method of OXC and metabolic product MHD and MHD-G in a kind of Simultaneously test human plasma, belong to technical field of biological.
Background technology
Epilepsy (Epilepsy) is the nervous system disease common in a kind of world wide, its Clinical symptoms is the cerebral function imbalance that the paradoxical discharge of cranial nerve cell recurrent exerbation causes, because its course of disease is long, disability rate is high, becomes the disease that the incidence of disease in paediatrics nervous system is the highest, harm is the most serious.As can not be well controlled, irreversible damage will be caused to cranial nerve, have a strong impact on growing of children, bring heavy psychological burden and economic pressures to infant, family and society.
Oxcarbazepine (Oxcarbazepine, OXC) is the medicine of a kind of novel treatment epilepsy partial seizures and generalized tonic-clonic outbreak.After Oxcarbazepine (OXC) enters body, liver cell solute enzyme (AKR1C1, AKR1C2, AKR1C3, AKR1C4, CBR1 and CBR3) active metabolite 10 can be translated into rapidly, 11-dihydro, 10-hydroxyl carbamazepine (MHD), and play pharmacological action, MHD is combined with glucuronic acid further again and excretes under UGT catalysis, fraction MHD is oxidized to 10 of parmacodynamics-less activity, 11-dihydroxy carbamazepine (DHD), the medicine of more than 90% is discharged by urine with metabolite form: glucuronide conjugate (49%), MHD (27%), DHD (3%), other metabolic products (13%), OXC is less than 1%, discharge from ight soil less than 4% former medicine.As can be seen here, metabolism is the principal mode eliminated in OXC body, II of MHD and UGT combines reaction, that OXC carries out the most important approach of biology conversion in vivo, affect UGT activity and will disturb MHD internal metabolism, thus change MHD blood concentration, cause Endodontic failure even to cause serious adverse reaction, high risks will be caused safely to patient medication.
Efficient liquid phase-tandem mass spectrometry (HPLC-MS/MS) is the most frequently used method of detection of drugs concentration, has the features such as highly sensitive, high specificity, can detect lot of trace compound simultaneously.Therefore, by while human body in the blood concentration of OXC, MHD, MHD-G, OXC dynamic metabolic alterations situation in vivo can be monitored more accurately, and judge whether to reduce due to OXC metabolism or UGT gene mutation causes the reduction of MHD medicine concentration.
At present, HPLC-MS/MS method through system verification can only detect OXC and MHD simultaneously, but also there is defect: as longer in the time of losing shape (10min) (High-performanceliquidchromatography-tandemmassspectrome trymethodforsimultaneousquantificationofcarbamazepine, oxcarbazepine, andtheirmainmetabolitesinhumanserum.TherDrugMonit.2012, 34 (1): 53-58), disposal route complexity (extraction method) (Simultaneousquantitativeanalysisof
oxcarbazepineand10,11-dihydro-10-hydroxycarbamazepineinhumanplasmabyliquidchromatography-electrospraytandemmassspectrometry.JPharmBiomedAnal.2007，45(2):304-311)。Detection method for MHD-G does not find pertinent literature, also the bibliographical information simultaneously detecting OXC, MHD, MHD-G is had no, the present invention establishes the HPLC-MS/MS assay method in OXC, MHD, MHD-G human plasma first, and solid methodology basis has been established in the detection for the clinical blood concentration of OXC.
Summary of the invention
The object of the invention is to overcome above-mentioned weak point, the detection method of OXC and metabolic product MHD and MHD-G in a kind of Simultaneously test human plasma is provided.
According to technical scheme provided by the invention, the detection method of OXC and metabolic product MHD and MHD-G in a kind of Simultaneously test human plasma, step is:
(1) plasma sample pre-service: get plasma sample, add interior mark intrazepam, then add acetonitrile precipitation albumen, centrifugation gets supernatant, obtains pretreated plasma sample;
(2) chromatogram: step (1) pretreated plasma sample is carried out liquid chromatography separation: chromatographic column in chromatographic condition: PhenomenexkinetexXB-C18 post; Mobile phase: acetonitrile: water volume ratio is 50:50, containing mass concentration in water is the formic acid of 0.1%; Flow velocity: 250 ~ 300 μ L/min; Column temperature: 30 ~ 40 DEG C; Sample size: 10 μ L;
(3) mass spectrum: ion gun: electro-spray ionization ESI ion gun, mode scans: select reaction detection scanning SRM, spray voltage: 3500 ~ 4000V, sheath gas: 20 ~ 50psi, assisted gas: 10 ~ 20arb, capillary temperature: 350 DEG C, ion source temperature: 300 DEG C;
(4) calculate: adopt internal standard method, substitute into typical curve equation with the peak area ratio of OXC, MHD, MHD-G and interior mark intrazepam, calculate the blood concentration of OXC, MHD, MHD-G.
Step (1) is specially: get plasma sample 150 μ L, and add mark intrazepam solution 15 μ L in 1000ng/mL, then add acetonitrile 285 μ L, the centrifugal 10 ~ 15min of vortex mixing 5 ~ 10min, 13000rpm, gets supernatant after centrifugal.
Step (1) described plasma sample is the blood plasma containing OXC, MHD, MHD-G.
In step (2), chromatogram column length is 100mm, and internal diameter is 2.1mm, and packing material size is 2.6 μm.
Ion is detected specific as follows: OXC selective reaction detects ion [M+H] in step (3)
+m/z253.2 → m/z180.2CE:28eV; MHD selective reaction detects ion [M+H]
+m/z255.2 → m/z194.2CE:21eV; MHD-G selective reaction detects ion [M+H]
+m/z431.1 → m/z194.2CE:28eV; Interior mark intrazepam selective reaction detects ion [M+H]
+m/z282.3 → m/z236.2CE:22eV.
Beneficial effect of the present invention:
(1) preprocess method is easy: adopt a step precipitation of protein, be applicable to conventional sense.
(2) specificity is strong, PhenomenexkinetexXB-C18 chromatographic column is adopted to be separated, acetonitrile: the mixed liquor of water (containing mass concentration in water is the formic acid of 0.1%) is as mobile phase, isocratic elution, the retention time of OXC is about 1.24min, the retention time of MHD is about 1.07min, MHD-G retention time is about 1.00min, the retention time of interior mark intrazepam (Nitrazepam) is about 1.55min, four have good degree of separation, can complete mensuration in 3min.In addition, endogenous material does not disturb both mensuration.
(3) highly sensitive: in blood plasma OXC, MHD, MHD-G be minimumly quantitatively limited to 10,50,125ng/mL.
(4) detect fast: 3min completes the once mensuration of sample, and the time is short, be therefore applicable to the detection of large batch of biological sample.
(5) amount of samples is little: only need 150 μ L plasma samples, can determine actual concentrations.
(6) the inventive method is quick, accurate, highly sensitive, easy and simple to handle, for the determination of plasma concentration of OXC, MHD, MHD-G provides foundation.OXC, MHD, MHD-G plasma standard curve linear scope of this method is 10 ~ 750ng/mL, 50 ~ 10000ng/mL, 125 ~ 5000ng/mL, and in low QC day, day to day precision and accuracy be all less than 20%; In middle and high QC day, day to day precision and accuracy be all less than 15%.
Accompanying drawing explanation
Fig. 1 is blank human plasma mass spectrogram.
Fig. 2 is the mass spectrogram adding OXC, MHD, MHD-G and interior mark intrazepam (Nitrazepam) in blank human plasma.
Fig. 3 is the canonical plotting of human plasma OXC.
Fig. 4 is the canonical plotting of human plasma MHD.
Fig. 5 is the canonical plotting of human plasma MHD-G.
Embodiment
According to following embodiment, the present invention may be better understood, but those skilled in the art will readily understand, the content described by embodiment only for illustration of the present invention, and should can not limit the present invention described in detail in claims yet.
Embodiment: the mensuration of human plasma OXC, MHD and MHD-G concentration
Experiment material and instrument
Oxcarbazepine, purity >99.8%, lot number: 100657-201102, is purchased from National Institute for Food and Drugs Control; 10,11-dihydro-10-hydroxyl carbamazepine, purity >98%, lot number: D449135, is purchased from lark prestige company; 10,11-dihydro-10-hydroxyl carbamazepine glucuronide conjugate, purity >95.5%, lot number: MD114541501, is purchased from carbosynth company of Britain; Intrazepam, lot number: 9201, is purchased from national arcotic laboratory; Methyl alcohol, lot number: HX080973, acetonitrile, lot number: HX086245, is all purchased from CNW company; Formic acid, lot number: T20090811, is purchased from Chemical Reagent Co., Ltd., Sinopharm Group; Water is ultrapure water (Milli-Q water purification machine) self-control.
Liquid chromatographic system: ACCELAautosampler, ACCELA1250pump, ThermoFisher company of the U.S.; MS/MS system: the triple quadrupole rods tandem mass spectrometry instrument of TSQQuantumAccess type, is equipped with electro-spray ionization ESI ion gun, ThermoFisher company of the U.S.; Data acquisition: LCQUAN
tM2.6 softwares, ThermoFisher company of the U.S.; The miniature vortex mixed instrument of XW-80A, Instrument Factory of Shanghai Medical Univ.; Z233MK-2 supercentrifuge, German HERMLE company; BT25S type analysis balance, German Sartorius company.HGC-12A Nitrogen evaporator, Tianjin Heng Ao company.
Liquid matter condition
1, liquid phase chromatogram condition:
Chromatographic column: PhenomenexkinetexXB-C18 (2.6 μm, 2.1 × 100mm); Mobile phase: acetonitrile: water (containing mass concentration in water is the formic acid of 0.1%)=50:50 (V/V); Flow velocity: 200 ~ 300 μ L/min; Column temperature: 30 ~ 35 DEG C; Sample size: 10 μ L.
2, Mass Spectrometry Conditions:
Ion gun: electro-spray ionization ESI ion gun, mode scans: select reaction detection scanning (SRM), spray voltage: 3500 ~ 4000V, sheath gas: 20 ~ 50psi, assisted gas: 10 ~ 20arb, capillary temperature: 350 DEG C, ion source temperature: 300 DEG C.Detect ion: OXC selective reaction detects ion [M+H]
+m/z253.2 → m/z180.2CE:28eV; MHD selective reaction detects ion [M+H]
+m/z255.2 → m/z194.2CE:21eV; MHD-G selective reaction detects ion [M+H]
+m/z431.1 → m/z194.2CE:28eV; Interior mark intrazepam selective reaction detects ion [M+H]
+m/z282.3 → m/z236.2CE:22eV;
Experimentation
1, OXC, MHD, MHD-G and the preparation of interior mark intrazepam standard solution
It is a small amount of that precision takes OXC standard items, and be placed in volumetric flask, methyl alcohol is configured to 1mg/mLOXC storing solution.Precision measures appropriate OXC storing solution methyl alcohol and dilutes successively, be made into concentration be respectively 100,200,500,1000,2500,5000, the OXC standard solution of 7500ng/mL.
It is a small amount of that precision takes MHD standard items, and be placed in volumetric flask, methyl alcohol is configured to 1mg/mLMHD storing solution.Precision measures appropriate MHD storing solution methyl alcohol and dilutes successively, be made into concentration be respectively 500,1250,5000,20000,50000,80000, the MHD standard solution of 100000ng/mL.
It is a small amount of that precision takes MHD-G standard items, and be placed in volumetric flask, methyl alcohol is configured to 1mg/mLMHD-G storing solution.Precision measures appropriate MHD-G storing solution methyl alcohol and dilutes successively, be made into concentration be respectively 1250,2500,5000,10000,20000,40000, the MHD-G standard solution of 50000ng/mL.
It is a small amount of that precision takes interior mark intrazepam standard items, and be placed in volumetric flask, methyl alcohol is configured to 1mg/mL intrazepam storing solution.Precision measures appropriate storing solution methanol dilution, and being configured to concentration is 1000ng/mL intrazepam standard solution.
2, specificity
Get the blank human plasma of 150 μ L, add 300 μ L acetonitrile precipitation albumen, the centrifugal 10 ~ 15min of vortex mixing 5 ~ 10min, 13000rpm, gets supernatant 10 μ L and carries out LC-MS/MS analysis.Blank human plasma sample's mass spectrogram as shown in Figure 1.
Get 1.5mLEP pipe number to prop up, add each 15 μ L of OXC, MHD, MHD-G standard solution, after nitrogen dries up, add the blank human plasma of 150 μ L, mark intrazepam solution and 285 μ L acetonitrile precipitation albumen in 15 μ L1000ng/mL, centrifugal 10 ~ the 15min of vortex mixing 5 ~ 10min, 13000rpm, gets supernatant 10 μ L and carries out LC-MS/MS analysis.
The mass spectrogram of OXC, MHD, MHD-G and interior mark intrazepam (Nitrazepam) is added as shown in Figure 2 in blank human plasma.
Note: passage a:OXC selective reaction detects ion [M+H]
+m/z253.2 → m/z180.2CE:28eV; Passage b:MHD selective reaction detects ion [M+H]
+m/z255.2 → m/z194.2CE:21eV; Passage c: interior mark intrazepam selective reaction detects ion [M+H]
+m/z282.3 → m/z236.2CE:22eV; Passage d:MHD-G selective reaction detects ion [M+H]
+m/z431.1 → m/z194.2CE:28eV.
Result shows, under the LC-MS/MS condition that this experiment adopts, without assorted peak, interference is caused to detection material in blood plasma, the retention time of OXC is about 1.24min, the retention time of MHD is about 1.07min, MHD-G retention time is about 1.00min, and the retention time of interior mark intrazepam is about 1.55min, completes mensuration in 3min.OXC, MHD, MHD-G and interior mark intrazepam do not interfere with each other, and peak shape is good, and baseline is steady.
3, typical curve
Get 1.5mLEP pipe number to prop up, add OXC, MHD, MHD-G standard serial solution respectively, after nitrogen dries up, add the blank human plasmas of 150 μ L, be configured to concentration for (OXC:10,20,50,100,250,500,750ng/mL; MHD:50,125,500,2000,500,8000,10000ng/mL; MHD-G:125,250,500,1000,2000,4000,5000ng/mL) standard plasma containing drug.Add mark intrazepam solution in 15 μ L1000ng/mL again, 285 μ L acetonitrile precipitation albumen, the centrifugal 10 ~ 15min of vortex mixing 5 ~ 10min, 13000rpm, gets supernatant 10 μ L and carries out LC-MS/MS analysis.Calculate the peak area As of OXC, MHD, MHD-G and the ratio f (f=As/Ai) of interior mark peak area Ai respectively, with testing concentration c for horizontal ordinate, with f value for ordinate, with weighting (weight coefficient: 1/x
2) least square method carries out linear regression operation and obtain regression equation.
OXC：f＝0.0055663+0.00615469*c，r
2＝0.9985；
MHD：f＝0.0245203+0.000508853*c，r
2＝0.9953；
MHD-G：f＝-0.00106133+8.3264e-005*c,r
2＝0.9967。
OXC is good in 10 ~ 750ng/mL scope internal linear relation, is minimumly quantitatively limited to 10ng/mL.
MHD is good in 50 ~ 10000ng/mL scope internal linear relation, is minimumly quantitatively limited to 50ng/mL.
MHD-G is good in 125 ~ 5000ng/mL scope internal linear relation, is minimumly quantitatively limited to 125ng/mL.
Specific standards curve map as in Figure 3-5.
4, minimum quantitative limit (LLOQ)
Get 1.5mLEP pipe number to prop up, add OXC, MHD, MHD-G standard solution respectively, after nitrogen dries up, add the blank human plasma of 150 μ L, be configured to concentration for (OXC:10ng/mL; MHD:50ng/mL; MHD-G:125ng/mL) standard plasma containing drug.Add mark intrazepam solution and 285 μ L acetonitrile precipitation albumen in 15 μ L1000ng/mL again, the centrifugal 10 ~ 15min of vortex mixing 5 ~ 10min, 13000rpm, gets supernatant 10 μ L and carries out LC-MS/MS analysis.Establish 5 Duplicate Samples simultaneously.Calculate the peak area As of OXC, MHD, MHD-G and the ratio f of interior mark peak area Ai, substitute in the plasma standard curve on the same day measured concentration of trying to achieve containing OXC, MHD, MHD-G.Precision (RSD%) and the accuracy (RE%) of minimum quantitative limit (LLOQ) is calculated by measured concentration.
As seen from the results in Table 1, RSD% and RE% of OXC, MHD, MHD-G all within 15%, the lower limit of quantitation (LLOQ) of OXC, MHD, MHD-G is respectively 10,50,125ng/mL (S/N>10).
Table 1LC-MS/MS method measures the lower limit of quantitation (LLOQ) of OXC, MHD, MHD-G in human plasma
| OXC(10ng/mL) | RE％ | MHD(50ng/mL) | RE％ | MHD-G(125ng/mL) | RE％ | |
| 1 | 10.60 | 106.04 | 42.18 | 84.36 | 130.23 | 104.18 |
| 2 | 10.06 | 100.61 | 47.39 | 94.78 | 122.59 | 98.07 |
| 3 | 9.31 | 93.09 | 48.12 | 96.23 | 100.02 | 80.02 |
| 4 | 9.84 | 98.42 | 45.54 | 91.07 | 110.87 | 88.70 |
| 5 | 10.45 | 104.46 | 48.54 | 97.07 | 131.07 | 104.85 |
| Mean value | 10.05 | 46.35 | 118.96 | |||
| RSD％ | 5.11 | 5.61 | 11.21 |
5, preci-sion and accuracy
Be basic, normal, high Quality Control (QC) sample according to the preparation of human plasma typical curve method containing OXC (20,100,500ng/mL), MHD (125,2000,8000ng/mL), MHD-G (250,1000,4000ng/mL) plasma sample.Add mark intrazepam solution and 285 μ L acetonitrile precipitation albumen in 15 μ L1000ng/mL again, the centrifugal 10 ~ 15min of vortex mixing 5 ~ 10min, 13000rpm, gets supernatant 10 μ L and carries out LC-MS/MS analysis.For three days on end, each concentration every day 5 Duplicate Samples, calculate the peak area As of OXC, MHD, MHD-G and the ratio f of interior mark peak area Ai, substitute in the plasma standard curve on the same day measured concentration of trying to achieve containing OXC, MHD, MHD-G, calculated in a few days by measured concentration, day to day precision, be relative standard deviation (RSD%), the ratio of measured concentration and theoretical concentration is accuracy (RE%).
In low QC day from table 2-4, OXC, MHD, MHD-G, day to day precision and accuracy be all less than 20%; In middle and high QC day, day to day precision and accuracy be all less than 15%.
Table 2LC-MS/MS method measure OXC in human plasma in a few days, day to day precision and accuracy
Table 3LC-MS/MS method measure MHD in human plasma in a few days, day to day precision and accuracy
Table 4LC-MS/MS method measure MHD-G in human plasma in a few days, day to day precision and accuracy
6, the recovery
Get the blank centrifuge tube number of 1.5mL to prop up, add OXC, MHD and MHD-G standard solution each 15 μ L, the N of variable concentrations respectively
2after drying up, add again 150 μ L people blank plasmas be mixed with basic, normal, high three concentration standard plasma containing drug (OXC plasma concentration is respectively 20,100,500ng/mL, MHD plasma concentration is respectively 125,2000,8000ng/mL, MHD-G plasma concentration is respectively 250,1000,4000ng/mL).Add mark intrazepam solution and 285 μ L acetonitrile precipitation albumen in 15 μ L1000ng/mL again, the centrifugal 10 ~ 15min of vortex mixing 5 ~ 10min, 13000rpm, gets supernatant 10 μ L and carries out LC-MS/MS analysis.Obtain relative medicine peak area and interior mark peak area, be respectively As (H), Ai (H); Each concentration carries out 5 sample analyses.
Get the blank centrifuge tube number of 1.5mL to prop up, add after adding 150 μ L people blank plasmas and reenter 300 μ L acetonitrile precipitation albumen, after the centrifugal 10 ~ 15min of vortex mixing 5 ~ 10min, 13000rpm, for subsequent use.Separately get the blank centrifuge tube number of 1.5mL to prop up, add OXC, MHD and MHD-G Quality Control (QC) solution and 1000ng/mL interior mark intrazepam solution each 15 μ L, the N of variable concentrations respectively
2under dry up, often pipe adds blank plasma supernatant after above-mentioned 450 μ L acetonitrile centrifugal treating, vortex dissolves, be mixed with the reference substance solution of basic, normal, high three concentration, sample introduction is analyzed, obtain relative medicine peak area and interior mark peak area, be respectively As (D), Ai (D), each concentration carries out 5 sample analyses.Extraction recovery is calculated as follows=[As (H)/As (D) is average] × 100%.
Table 5-7 result shows, the OXC recovery is 85.48% ~ 96.24%, the MHD recovery be 90.73% ~ 97.95%, the MHD recovery is 72.30% ~ 80.63%.
Table 5LC-MS/MS method measures the recovery (n=5) of OXC in blood plasma
Table 6LC-MS/MS method measures the recovery (n=5) of MHD in blood plasma
Table 7LC-MS/MS method measures the recovery (n=5) of MHD-G in blood plasma
7, testing result
Detect after 10 infants take Oxcarbazepine (OXC), the Grain volume of blood plasma OXC, MHD, MHD-G, in table 8.
Table 8
| Sample | OXC(ng/mL) | MHD(ng/mL) | MHD-G(ng/mL) |
| Sample-1 | 54.20 | 3629.71 | 694.95 |
| Sample-2 | 124.30 | 5731.20 | 1607.33 |
| Sample-3 | 96.70 | 6055.96 | 1459.04 |
| Sample-4 | 55.26 | 4047.15 | 1286.54 |
| Sample-5 | 51.38 | 4339.84 | 507.61 |
| Sample-6 | 77.90 | 4360.72 | 578.98 |
| Sample-7 | 48.84 | 2826.03 | 856.97 |
| Sample-8 | 120.55 | 5892.58 | 619.28 |
| Sample-9 | 75.14 | 5244.91 | 614.33 |
| Sample-10 | 77.00 | 6466.42 | 1667.10 |
From table 1-8, the inventive method is quick, accurate, highly sensitive, easy and simple to handle, for the determination of plasma concentration of OXC, MHD, MHD-G provides foundation.OXC, MHD, MHD-G plasma standard curve linear scope of this method is 10 ~ 750ng/mL, 50 ~ 10000ng/mL, 125 ~ 5000ng/mL, and in low QC day, day to day precision and degree of accuracy be all less than 20%; In middle and high QC day, day to day precision and accuracy be all less than 15%.
Claims (5)
1. a detection method of OXC and metabolic product MHD and MHD-G in Simultaneously test human plasma, is characterized in that step is:
(1) plasma sample pre-service: get plasma sample, add interior mark intrazepam, then add acetonitrile precipitation albumen, centrifuging and taking supernatant, obtains plasma sample;
(2) chromatogram: step (1) pretreated plasma sample is carried out liquid chromatography separation: chromatographic column in chromatographic condition: PhenomenexkinetexXB-C18 post; Mobile phase: acetonitrile: water volume ratio is 50:50, containing mass concentration in water is the formic acid of 0.1%; Flow velocity: 250 ~ 300 μ L/min; Column temperature: 30 ~ 40 DEG C; Sample size: 10 μ L;
(3) mass spectrum: ion gun: electro-spray ionization ESI ion gun, scan mode: select reaction detection scanning SRM, spray voltage: 3500 ~ 4000V, sheath gas: 20 ~ 50psi, assisted gas: 10 ~ 20arb, capillary temperature: 350 DEG C, ion source temperature: 300 DEG C;
(4) calculate: adopt internal standard method, substitute into typical curve equation with the peak area ratio of OXC, MHD and MHD-G and interior mark intrazepam, calculate the blood concentration of OXC, MHD and MHD-G.
2. the detection method of OXC and metabolic product MHD and MHD-G in Simultaneously test human plasma as claimed in claim 1, it is characterized in that step (1) is specially: get plasma sample 150 μ L, add mark intrazepam solution 15 μ L in 1000ng/mL, add acetonitrile 285 μ L again, vortex mixing 5 ~ 10min, centrifugal 10 ~ the 15min of 13000rpm, gets supernatant after centrifugal.
3. the detection method of OXC and metabolic product MHD and MHD-G in Simultaneously test human plasma as claimed in claim 1, is characterized in that: step (1) described plasma sample is the blood plasma containing OXC, MHD and MHD-G.
4. the detection method of OXC and metabolic product MHD and MHD-G in Simultaneously test human plasma as claimed in claim 1, it is characterized in that in step (2), chromatogram column length is 100mm, internal diameter is 2.1mm, and packing material size is 2.6 μm.
5. the detection method of OXC and metabolic product MHD and MHD-G in Simultaneously test human plasma as claimed in claim 1, is characterized in that detecting ion in step (3) specific as follows: OXC selective reaction detects ion [M+H]
+m/z253.2 → m/z180.2CE:28eV; MHD selective reaction detects ion [M+H]
+m/z255.2 → m/z194.2CE:21eV; MHD-G selective reaction detects ion [M+H]
+m/z431.1 → m/z194.2CE:28eV; Interior mark intrazepam selective reaction detects ion [M+H]
+m/z282.3 → m/z236.2CE:22eV.
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| CN112034078A (en) * | 2020-09-21 | 2020-12-04 | 北京和合医学诊断技术股份有限公司 | Method for detecting carbamazepine |
| CN112946100A (en) * | 2021-01-26 | 2021-06-11 | 山东英盛生物技术有限公司 | Method and kit for determining carbamazepine and 10-hydroxycarbazepine and application thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108195978A (en) * | 2018-03-30 | 2018-06-22 | 北京和合医学诊断技术股份有限公司 | Detect Oxcarbazepine and its liquid phase chromatography analytical method of metabolite blood concentration in blood |
| CN109541107A (en) * | 2018-11-30 | 2019-03-29 | 徐州佳生医药科技有限公司 | A kind of method that LC-MS measures Carbamazepine in blood plasma |
| CN109884235A (en) * | 2019-02-28 | 2019-06-14 | 上海药明康德新药开发有限公司 | The efficient liquid phase detection method of carbamazepine |
| CN110161159A (en) * | 2019-07-04 | 2019-08-23 | 杭州必益泰得医学科技有限公司 | A kind of biological sample analysis method of Oxcarbazepine bioequivalence test |
| CN110763800A (en) * | 2019-11-12 | 2020-02-07 | 北京和合医学诊断技术股份有限公司 | Method for detecting oxcarbazepine and 10, 11-dihydro-10-hydroxycarbazepine in blood |
| CN112034078A (en) * | 2020-09-21 | 2020-12-04 | 北京和合医学诊断技术股份有限公司 | Method for detecting carbamazepine |
| CN114671809A (en) * | 2020-12-25 | 2022-06-28 | 苏州博源医疗科技有限公司 | Oxcarbazepine derivative, immunogen, anti-oxcarbazepine specific antibody, preparation method and application thereof |
| CN112946100A (en) * | 2021-01-26 | 2021-06-11 | 山东英盛生物技术有限公司 | Method and kit for determining carbamazepine and 10-hydroxycarbazepine and application thereof |
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