CN105136957B - 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 OXC and the detection method of metabolite MHD and MHD-G in a kind of human plasma of mensuration simultaneously, belong to
In technical field of biological.
Background technology
Epilepsy (Epilepsy) is nervous system disease common in a kind of world wide, and its Clinical symptoms is that cranial nerve is thin
The brain function imbalance that the paradoxical discharge of born of the same parents' recurrent exerbation causes, because its course of disease is long, disability rate is high, becomes in department of pediatrics nervous system
The disease that sickness rate is the highest, harm is the most serious.As can not be well controlled, it will cranial nerve to be caused irreversible damage,
Have a strong impact on the growth promoter of child, bring heavy psychological burden and economic pressures to infant, family and society.
Oxcarbazepine (Oxcarbazepine, OXC) is a kind of novel treatment epilepsy partial seizures and generalized tonic-clonic
Property outbreak medicine.After oxcarbazepine (OXC) enters body, hepatocyte solute enzyme (AKR1C1, AKR1C2, AKR1C3,
AKR1C4, CBR1 and CBR3) active metabolite 10,11-dihydro, 10-hydroxyl carbamazepine can be translated into rapidly
(MHD), and playing pharmacological action, MHD is further combined with glucuronic acid under UGT is catalyzed and excretes, fraction MHD
Being oxidized to the 10 of parmacodynamics-less activity, 11-dihydroxy carbamazepine (DHD), the medicine of more than 90% leads to metabolite form
Cross urine to discharge: glucuronide conjugate (49%), MHD (27%), DHD (3%), other metabolites (13%), OXC is few
In 1%；Discharge from feces less than 4% former medicine.As can be seen here, metabolism is the interior principal mode eliminated of OXC body, MHD's Yu UGT
II combines reaction, is that OXC carries out the most important approach of bioconversion in vivo, affects UGT activity and will disturb generation in MHD body
Thank, thus change MHD blood drug level, cause Endodontic failure even to cause serious adverse reaction, patient medication will be caused safely
High risks.
Efficiently liquid phase-tandem mass spectrometry (HPLC-MS/MS) is detection drug level most common method, has sensitivity
The features such as height, high specificity, can detect lot of trace compound simultaneously.Therefore, by detecting in human body simultaneously
The blood drug level of OXC, MHD, MHD-G, can monitor OXC dynamic metabolic alterations situation in vivo more accurately, and sentence
Disconnected whether reduce due to OXC metabolism or UGT gene mutation causes the reduction of MHD medicine concentration.
At present, the HPLC-MS/MS method through system checking can only detect OXC and MHD simultaneously, but also have the disadvantage that as
Time of losing shape longer (10min) (High-performance liquid chromatography-tandem mass
spectrometry method for simultaneousquantification ofcarbamazepine,
Oxcarbazepine, and their main metabolites in human serum.Ther Drug Monit.2012,
34 (1): 53-58), complicated (extraction method) (Simultaneous quantitative analysis of of processing method
oxcarbazepine and 10,11-dihydro-10-hydroxycarbamazepine inhuman
plasma by liquid chromatography-electrospray tandem mass spectrometry.J Pharm
Biomed Anal.2007,45 (2): 304-311).Detection method for MHD-G does not finds pertinent literature, has no simultaneously yet
The document report of detection OXC, MHD, MHD-G, the present invention establishes the HPLC-MS/MS in OXC, MHD, MHD-G human plasma first
Assay method, solid methodology basis has been established in the detection for the clinical blood drug level of OXC.
Summary of the invention
It is an object of the invention to overcome above-mentioned weak point, it is provided that one measures OXC and metabolism in human plasma simultaneously
The detection method of product MHD and MHD-G.
The technical scheme provided according to the present invention, one measures OXC and metabolite MHD and MHD-in human plasma simultaneously
The detection method of G, step is:
(1) plasma sample pretreatment: take plasma sample, adds internal standard nitrodiazepam, adds acetonitrile precipitation albumen, speed
Centrifuging and taking supernatant, obtains pretreated plasma sample；
(2) chromatograph: step (1) pretreated plasma sample is carried out liquid chromatograph separation: chromatographic column in chromatographic condition:
Phenomenex kinetex XB-C18 post；Flowing phase: acetonitrile: water volume ratio is 50:50, water containing mass concentration 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 source: electro-spray ionization ESI ion source, mode scans: selective response detection scanning SRM, spray
Mist voltage: 3500～4000V, sheath gas: 20～50psi, auxiliary gas: 10～20arb, capillary temperature: 350 DEG C, ion source temperature
Degree: 300 DEG C；
(4) calculate: use internal standard method, substitute into standard with the peak area ratio of OXC, MHD, MHD-G and internal standard nitrodiazepam
Curvilinear equation, calculates the blood drug level of OXC, MHD, MHD-G.
Step (1), particularly as follows: take plasma sample 150 μ L, adds 1000ng/mL internal standard nitrodiazepam solution 15 μ L, then adds
Entering acetonitrile 285 μ L, vortex mixing 5～10min, 13000rpm is centrifuged 10～15min, takes supernatant after being centrifuged.
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.
In step (3), detection ion is specific as follows: OXC selective reaction detection ion [M+H]+m/z253.2→m/
z180.2CE:28eV；MHD selective reaction detection ion [M+H]+m/z 255.2→m/z194.2CE:21eV；MHD-G selects
Property reaction detection ion [M+H]+m/z 431.1→m/z194.2CE:28eV；Internal standard nitrodiazepam selective reaction detection ion
[M+H]+m/z 282.3→m/z 236.2CE:22eV。
Beneficial effects of the present invention:
(1) preprocess method is easy: use a step precipitation of protein, it is adaptable to conventional sense.
(2) specificity is strong, uses Phenomenex kinetex XB-C18 chromatographic column to separate, acetonitrile: water is (in water
Be the formic acid of 0.1% containing mass concentration) mixed liquor as flowing phase, isocratic elution, the retention time of OXC be about
The retention time of 1.24min, MHD is about 1.07min, MHD-G retention time and is about 1.00min, internal standard nitrodiazepam
(Nitrazepam) retention time is about 1.55min, and four have good separating degree, can complete to measure in 3min.This
Outward, endogenous material does not disturb both mensuration.
(3) highly sensitive: in blood plasma OXC, MHD, MHD-G minimum be quantitatively limited to 10,50,125ng/mL.
(4) detection is quickly: 3min completes the once mensuration of sample, and the time is short, is therefore suitable for large batch of biological sample
Detection.
(5) amount of samples is little: only needs 150 μ L plasma samples, can determine actual concentrations.
(6) the inventive method is quick, accurate, highly sensitive, easy and simple to handle, and the blood drug level for OXC, MHD, MHD-G is surveyed
Foundation is provided surely.OXC, MHD, MHD-G plasma standard curve linear scope of this method be 10～750ng/mL, 50～
10000ng/mL, 125～5000ng/mL, in low QC day, day to day precision and accuracy be respectively less than 20%；In middle and high QC day,
Day to day precision and accuracy are respectively less than 15%.
Accompanying drawing explanation
Fig. 1 is blank human plasma mass spectrum.
Fig. 2 is to add OXC, MHD, MHD-G and the mass spectrum of internal standard nitrodiazepam (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.
Detailed description of the invention
According to following embodiment, the present invention be may be better understood, but as it will be easily appreciated by one skilled in the art that enforcement
Content described by example is merely to illustrate the present invention, and should be also without limitation on described in detail this in claims
Bright.
Embodiment: the mensuration of human plasma OXC, MHD and MHD-G concentration
Experiment material and instrument
Oxcarbazepine, purity > 99.8%, lot number: 100657-201102, it is purchased from National Institute for Food and Drugs Control；
10,11-dihydro-10-hydroxyl carbamazepines, purity > 98%, lot number: D449135, it is purchased from lark prestige company；10,11-dihydro-
10-hydroxyl carbamazepine glucuronide conjugate, purity > 95.5%, lot number: MD114541501, it is purchased from Britain
Carbosynth company；Nitrodiazepam, lot number: 9201, it is purchased from country's narcotic laboratory；Methanol, lot number: HX080973, second
Nitrile, lot number: HX086245, all it is purchased from CNW company；Formic acid, lot number: T20090811, it is purchased from the traditional Chinese medicines group limited public affairs of chemical reagent
Department；Water is ultra-pure water (Milli-Q water purification machine) self-control.
Liquid chromatographic system: ACCELA autosampler, ACCELA 1250pump, U.S. Thermo Fisher is public
Department；MS/MS system: TSQ QuantumAccess type triple quadrupole rods tandem mass spectrometry instrument, is equipped with electro-spray ionization ESI ion
Source, Thermo Fisher company of the U.S.；Data acquisition: LC QUANTM2.6 softwares, Thermo Fisher company of the U.S.；XW-
80A miniature vortex mixed instrument, Instrument Factory of Shanghai Medical Univ.；Z233MK-2 high speed centrifuge, HERMLE company of Germany；BT25S type divides
Analysis balance, Sartorius company of Germany.HGC-12A Nitrogen evaporator, Tianjin Heng Ao company.
Liquid matter condition
1, liquid phase chromatogram condition:
Chromatographic column: Phenomenex kinetex XB-C18 (2.6 μm, 2.1 × 100mm)；Flowing phase: acetonitrile: water (water
In containing mass concentration be the formic acid of 0.1%)=50:50 (V/V)；Flow velocity: 200～300 μ L/min；Column temperature: 30～35 DEG C；Enter
Sample amount: 10 μ L.
2, Mass Spectrometry Conditions:
Ion source: electro-spray ionization ESI ion source, mode scans: selective response detection scanning (SRM), spray voltage:
3500～4000V, sheath gas: 20～50psi, auxiliary gas: 10～20arb, capillary temperature: 350 DEG C, ion source temperature: 300
℃.Detection ion: OXC selective reaction detection ion [M+H]+m/z 253.2→m/z180.2CE:28eV；MHD selectivity is anti-
Ion [M+H] should be detected+m/z 255.2→m/z194.2CE:21eV；MHD-G selective reaction detection ion [M+H]+m/z
431.1→m/z194.2CE:28eV；Internal standard nitrodiazepam selective reaction detection ion [M+H]+m/z 282.3→m/z
236.2CE:22eV；
Experimentation
1, OXC, MHD, MHD-G and the preparation of internal standard nitrodiazepam standard solution
It is a small amount of that precision weighs OXC standard substance, is placed in volumetric flask, and methanol is configured to 1mg/mL OXC storing solution.Accurate amount
Take appropriate OXC storing solution methanol to dilute 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 weighs MHD standard substance, is placed in volumetric flask, and methanol is configured to 1mg/mL MHD storing solution.Accurate amount
Take appropriate MHD storing solution methanol to dilute 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 weighs MHD-G standard substance, is placed in volumetric flask, and methanol is configured to 1mg/mL MHD-G storing solution.Essence
Close measure appropriate MHD-G storing solution methanol and dilute 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 weighs internal standard nitrodiazepam standard substance, is placed in volumetric flask, and methanol is configured to 1mg/mL nitrodiazepam
Storing solution.Precision measures appropriate storing solution methanol dilution, and being configured to concentration is 1000ng/mL nitrodiazepam standard solution.
2, specificity
Taking 150 μ L blank human plasmas, add 300 μ L acetonitrile precipitation albumen, vortex mixing 5～10min, 13000rpm is centrifuged
10～15min, take supernatant 10 μ L and carry out LC-MS/MS analysis.Blank human plasma sample's mass spectrum is as shown in Figure 1.
Take 1.5mL EP pipe number to prop up, add each 15 μ L of OXC, MHD, MHD-G standard solution, after nitrogen dries up, add 150 μ L
Blank human plasma, 15 μ L 1000ng/mL internal standard nitrodiazepam solution and 285 μ L acetonitrile precipitation albumen, vortex mixing 5～
10min, 13000rpm are centrifuged 10～15min, take supernatant 10 μ L and carry out LC-MS/MS analysis.
Blank human plasma adds OXC, MHD, MHD-G and mass spectrum such as Fig. 2 of internal standard nitrodiazepam (Nitrazepam)
Shown in.
Note: passage a:OXC selective reaction detection ion [M+H]+m/z 253.2→m/z180.2CE:28eV；Passage b:
MHD selective reaction detection ion [M+H]+m/z 255.2→m/z194.2CE:21eV；Passage c: internal standard nitrodiazepam selects
Property reaction detection ion [M+H]+m/z 282.3→m/z 236.2CE:22eV；Passage d:MHD-G selective reaction detection ion
[M+H]+m/z 431.1→m/z194.2CE:28eV。
Result shows, under the conditions of the LC-MS/MS that this experiment is used, causes dry without miscellaneous peak to detection material in blood plasma
Disturbing, the retention time of OXC is about the retention time of 1.24min, MHD and is about 1.07min, MHD-G retention time about
1.00min, the retention time of internal standard nitrodiazepam is about 1.55min, completes to measure in 3min.OXC, MHD, MHD-G and internal standard
Nitrodiazepam does not interfere with each other, and peak shape is good, and baseline is steady.
3, standard curve
Take 1.5mL EP pipe number to prop up, be separately added into OXC, MHD, MHD-G standard serial solution, after nitrogen dries up, add 150
μ L blank human plasma, 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 contain
Medicine blood plasma.Add 15 μ L1000ng/mL internal standard nitrodiazepam solution, 285 μ L acetonitrile precipitation albumen, vortex mixing 5～
10min, 13000rpm are centrifuged 10～15min, take supernatant 10 μ L and carry out LC-MS/MS analysis.Calculate OXC, MHD, MHD-respectively
The ratio f (f=As/Ai) of the peak area As and internal standard peak area Ai of G, with testing concentration c as abscissa, sits with f value for vertical
Mark, with weighting (weight coefficient: 1/x2) method of least square carries out linear regression operation and obtain regression equation.
OXC:f=0.0055663+0.00615469*c, r2=0.9985；
MHD:f=0.0245203+0.000508853*c, r2=0.9953；
MHD-G:f=-0.00106133+8.3264e-005*c, r2=0.9967.
OXC linear relationship in the range of 10～750ng/mL is good, minimum is quantitatively limited to 10ng/mL.
MHD linear relationship in the range of 50～10000ng/mL is good, minimum is quantitatively limited to 50ng/mL.
MHD-G linear relationship in the range of 125～5000ng/mL is good, minimum is quantitatively limited to 125ng/mL.
Specific standards curve chart is as in Figure 3-5.
4, minimum quantitative limit (LLOQ)
Take 1.5mL EP pipe number to prop up, be separately added into OXC, MHD, MHD-G standard solution, after nitrogen dries up, add 150 μ L empty
White man's blood plasma, is configured to concentration for (OXC:10ng/mL；MHD:50ng/mL；MHD-G:125ng/mL) standard plasma containing drug.
Adding 15 μ L 1000ng/mL internal standard nitrodiazepam solution and 285 μ L acetonitrile precipitation albumen, vortex mixes 5～10min,
13000rpm is centrifuged 10～15min, takes supernatant 10 μ L and carries out LC-MS/MS analysis.Set 5 Duplicate Samples simultaneously.Calculating OXC,
The peak area As and the ratio f of internal standard peak area Ai of MHD, MHD-G, substitute into the same day plasma standard curve in try to achieve containing OXC,
The measured concentration of MHD, MHD-G.Precision (RSD%) and the accuracy of minimum quantitative limit (LLOQ) is calculated by measured concentration
(RE%).
As seen from the results in Table 1, RSD% and RE% of OXC, MHD, MHD-G all within 15%, OXC, MHD, MHD-G's
Lower limit of quantitation (LLOQ) be 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 |
| Meansigma methods | 10.05 | 46.35 | 118.96 | |||
| RSD% | 5.11 | 5.61 | 11.21 |
5, preci-sion and accuracy
According to the preparation of human plasma standard curve method containing OXC (20,100,500ng/mL), MHD (125,2000,8000ng/
ML), MHD-G (250,1000,4000ng/mL) plasma sample is basic, normal, high Quality Control (QC) sample.Add 15 μ L
1000ng/mL internal standard nitrodiazepam solution and 285 μ L acetonitrile precipitation albumen, vortex mixing 5～10min, 13000rpm is centrifuged
10～15min, take supernatant 10 μ L and carry out LC-MS/MS analysis.For three days on end, every day 5 Duplicate Samples of each concentration, calculate OXC,
The peak area As and the ratio f of internal standard peak area Ai of MHD, MHD-G, substitute into the same day plasma standard curve in try to achieve containing OXC,
The measured concentration of MHD, MHD-G, is calculated in a few days by measured concentration, day to day precision, is relative standard deviation (RSD%), real
The ratio surveying concentration and theoretical concentration is accuracy (RE%).
From table 2-4, in the low QC day of OXC, MHD, MHD-G, day to day precision and accuracy be respectively less than 20%；In,
In high QC day, day to day precision and accuracy be respectively 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 response rate
Take 1.5mL blank centrifuge tube number to prop up, be separately added into each 15 μ L of OXC, MHD and MHD-G standard solution of variable concentrations,
N2After drying up, add 150 μ L people's blank plasmas and be configured to the standard plasma containing drug of basic, normal, high three concentration (OXC blood plasma is dense
Degree is respectively 20,100,500ng/mL, MHD plasma concentration be respectively 125,2000,8000ng/mL, MHD-G plasma concentration respectively
Be 250,1000,4000ng/mL).Add 15 μ L 1000ng/mL internal standard nitrodiazepam solution and 285 μ L acetonitrile precipitation eggs
In vain, vortex mixing 5～10min, 13000rpm is centrifuged 10～15min, takes supernatant 10 μ L and carries out LC-MS/MS analysis.Obtain phase
Answer medicine peak area and internal standard peak area, respectively As (H), Ai (H)；Each concentration carries out 5 sample analyses.
Take 1.5mL blank centrifuge tube number to prop up, add after adding 150 μ L people's blank plasmas and reenter 300 μ L acetonitrile precipitation albumen, whirlpool
Rotation mixing 5～10min, after 13000rpm is centrifuged 10～15min, standby.Separately take 1.5mL blank centrifuge tube number to prop up, be separately added into
OXC, MHD and MHD-G Quality Control (QC) solution of variable concentrations and each 15 μ L of 1000ng/mL internal standard nitrodiazepam solution, N2Under blow
Dry, blank plasma supernatant after often pipe adds above-mentioned 450 μ L acetonitrile centrifugal treating, vortex dissolves, is configured to basic, normal, high three
The reference substance solution of concentration, sample introduction analysis, obtain relative medicine peak area and internal standard peak area, respectively As (D), Ai (D), often
One 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 response rate is 85.48%～96.24%, the MHD response rate be 90.73%～
97.95%, the MHD response rate is 72.30%～80.63%.
Table 5LC-MS/MS method measures the response rate (n=5) of OXC in blood plasma
Table 6LC-MS/MS method measures the response rate (n=5) of MHD in blood plasma
Table 7LC-MS/MS method measures the response rate (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, it is shown 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 blood of OXC, MHD, MHD-G
Concentration measures provides foundation.OXC, MHD, MHD-G plasma standard curve linear scope of this method is 10～750ng/mL, 50
～10000ng/mL, 125～5000ng/mL, in low QC day, day to day precision and degree of accuracy be respectively less than 20%；Middle and high QC day
In, day to day precision and accuracy be respectively less than 15%.
Claims (1)
1. measure OXC and a detection method of metabolite MHD and MHD-G in human plasma simultaneously, it is characterized in that step is:
(1) plasma sample pretreatment: take plasma sample, adds internal standard nitrodiazepam, adds acetonitrile precipitation albumen, in centrifuging and taking
Clear liquid, obtains plasma sample；
(2) chromatograph: step (1) pretreated plasma sample is carried out liquid chromatograph separation: chromatographic column in chromatographic condition:
Phenomenex kinetex XB-C18 post；Flowing phase: acetonitrile: water volume ratio is 50:50, water containing mass concentration 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 source: electro-spray ionization ESI ion source, scan mode: selective response detection scanning SRM, spraying electricity
Pressure: 3500 ~ 4000V, sheath gas: 20 ~ 50psi, assists gas: 10 ~ 20arb, capillary temperature: 350 DEG C, ion source temperature: 300
℃；
(4) calculate: use internal standard method, substitute into standard curve with the peak area ratio of OXC, MHD and MHD-G and internal standard nitrodiazepam
Equation, calculates the blood drug level of OXC, MHD and MHD-G；
Step (1), particularly as follows: take plasma sample 150 μ L, adds 1000ng/mL internal standard nitrodiazepam solution 15 μ L, adds second
Nitrile 285 μ L, vortex mixing 5 ~ 10min, 13000rpm are centrifuged 10 ~ 15min, take supernatant after being centrifuged；Step (1) described blood plasma sample
Product are the blood plasma containing OXC, MHD and MHD-G；
In step (2), chromatogram column length is 100mm, and internal diameter is 2.1mm, and packing material size is 2.6 μm；
In step (3), detection ion is specific as follows: OXC selective reaction detection ion [M+H]+ m/z 253.2→ m/z180.2
CE: 28eV；MHD selective reaction detection ion [M+H]+m/z 255.2→ m/z194.2 CE: 21eV；MHD-G selects
Property reaction detection ion [M+H]+m/z 431.1→ m/z194.2 CE: 28eV；Internal standard nitrodiazepam selective reaction detects
Ion [M+H]+ m/z 282.3 → m/z 236.2 CE: 22eV。
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| 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 |
| CN110161159B (en) * | 2019-07-04 | 2020-09-22 | 杭州必益泰得医学科技有限公司 | Biological sample analysis method for 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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