CN106645492B - Under the conditions of a kind of real-time detection fume exposure in animal brain monoamine neurotransmitter variation method - Google Patents
Under the conditions of a kind of real-time detection fume exposure in animal brain monoamine neurotransmitter variation method Download PDFInfo
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Abstract
Under the conditions of a kind of real-time detection fume exposure in animal brain monoamine neurotransmitter variation method, it is characterized by: animal is exposed in flue gas in real time using animal mouth and nose formula sucking fume exposure system, animal brain's sample is collected by on-line microdialysis technology, recycle high performance liquid chromatography-coulomb array electrochemical detector in brain dialysate samples monoamine neurotransmitter and its metabolin quickly analyzed, thus in the animal brain of real-time monitoring fume exposure front and back monoamine neurotransmitter variation characteristic.This method is closer to the mode of mankind's smoking tobaccos product to the fume exposure of animal, overcome can not simulate in real time, evaluation of flue gas sucking (nicotine intake) caused by monoamine neurotransmitter variation technical problem, have many advantages, such as that easy to operate, result is accurate and reliable, obvious progress is achieved compared with prior art, has started a kind of new monitoring method for Monoamine Metabolism variation in animal brain after fume exposure.
Description
Technical field
The present invention relates to the method for monoamine neurotransmitter mutation analysis in animal brain under the conditions of real-time detection fume exposure,
This method collects the experimental animal brain tissue sample being exposed in flue gas in real time by on-line microdialysis technology, recycles efficient liquid
Phase chromatography-coulomb array electrochemical detector in brain dialysate samples monoamine neurotransmitter and its metabolin carry out it is quick
On-line analysis, thus Real-Time Evaluation flue gas sucking exposure front and back monoamine neurotransmitter variation characteristic.
Background technique
Cigarette smoke and electronics smoke sol all important feature ingredients containing tobacco --- nicotine (Nicotine).Nicotine
The main reason for dependence is smoking addiction, and continue smoking tobaccos product and then endanger the basic reason of health.It takes the photograph
Entering intracorporal nicotine through blood-brain barrier and can be distributed widely in each region of brain tissue rapidly, and it is more to mainly act on middle limbic brain
Bar amine (Dopamine, DA) system and generate rewarding effect.In fact, including the monoamines such as DA, serotonin (5-HT) nerve
Mediator has been involved in the generating process of nicotine Neuropharmacology effect.Therefore, developing the new detection method of one kind can be to flue gas
The release of monoamine neurotransmitter and metabolic alterations carry out Accurate Determining and Real-Time Evaluation in animal brain caused by exposure, will be to take off
Show that the genesis mechanism of smoking addiction provides important means support.
The measurement of monoamine neurotransmitter mostly uses HPLC-electrochemical detection (HPLC- in biological sample
ECD), wherein high performance liquid chromatography-coulomb array electrochemical detector because its multichannel in response to due to show highly selective and
High sensitivity, application are relatively broad.It is combined using on-line microdialysis technology (Online microdialysis) and HPLC-ECD
The living body to monoamine neurotransmitters such as DA, 5-HT, dynamic measurement may be implemented, but this method is only capable of evaluation drug exposure
The variation of monoamine neurotransmitters caused by after (such as nicotine injection)；Using animal, integrally exposure sucks flue gas (by animal
Be placed in closed flue gas cabin) exposure chamber can not also synchronously complete that flue gas is exposed in real time and animal brain in monoamine nerve pass
The on-line determination of matter.
Summary of the invention
The present invention is exactly to be directed to not synchronizing at present to realize flue gas sucking exposure and monoamine neurotransmitter on-line determination
Deficiency, and the analysis side that monoamine neurotransmitter changes in animal brain under the conditions of a kind of real-time monitoring fume exposure specially developed
Method.This method realizes animal mouth and nose formula fume exposure system by sucking exposure clamper using improved experimental animal nose
The combination of system and Microdialysis sgstem, and then detected by using on-line microdialysis and high performance liquid chromatography-coulomb array electrochemical
Device joint technology realizes real-time acquisition and the on-line determination that exposure experimental animal brain tissue sample is sucked to flue gas, has reached cigarette
Aspiration enters the technical requirements of exposed progress synchronous with the variation measurement of intracerebral monoamine neurotransmitter.In the method, experimental animal
It is exposed in flue gas in such a way that only mouth and nose suck (nose-only), is closer to the side of mankind's smoking tobaccos product
Formula, overcome can not Real-Time Evaluation flue gas sucking (nicotine intake) caused by monoamine neurotransmitter variation technical problem, have
Easy to operate, the advantages that result is accurate and reliable.
The purpose of the present invention is achieved through the following technical solutions: animal under the conditions of a kind of real-time detection fume exposure
Animal is exposed to cigarette using animal mouth and nose formula sucking fume exposure system by the method for intracerebral monoamine neurotransmitter variation in real time
In gas, animal brain's sample is collected by on-line microdialysis technology, recycles high performance liquid chromatography-coulomb array electrochemical inspection
Survey device in brain dialysate samples monoamine neurotransmitter and its metabolin quickly analyzed, so that real-time monitoring flue gas is sudden and violent
The variation characteristic of monoamine neurotransmitter in the animal brain of dew front and back, and then according to the analysis of DA, 5-HT in sample and its metabolin
Intracerebral monoamine neurotransmitter changing rule caused by testing result Real-Time Evaluation fume exposure.
Specific step is as follows for this method:
A. fume exposure: making electronic cigarette liquid generate aerosol using smoking machine smoking cigarette or using aerosol generator,
Head is put into a dedicated experimental animal fixing clamp holder equipped with the animal of microdialysis probe conduit, so that the clamper is held under the arm
It holds animal and is only exposed in certain density flue gas with mouth and nose suction；
B. encephalic micro-dialysis sample collection: will be in the conduit of microdialysis probe insertion animal head after operation Microdialysis sgstem
Continuous animal brain dialyzate of collecting is automatically injected by online sample injector at regular intervals in the quantitative loop of online sample injector
In the high performance liquid chromatography of operation.
C. high performance liquid chromatography-coulomb array electrochemical detector analysis: setting gradient elution program and sample introduction sequence are simultaneously
It runs high performance liquid chromatography-coulomb array electrochemical detector and detects microdialysis sample, so that the detection time of each sample is omited
Less than the acquisition time of the sample.
D. standard solution is prepared and Specification Curve of Increasing: weighing DA, 3,4 dihydroxyphenyl acetic acids (DOPAC), high-quality slender joss stick respectively
5 kinds of oxalic acid (HVA), 5-HT and 5 hydroxyindole acetic acids (5-HIAA) each 10mg of standard items, are respectively placed in 100mL volumetric flask, use
Methanol/water solution (20:80, volume ratio, similarly hereinafter) dissolves and is diluted to scale, shakes up, the standard that obtain concentration respectively be 100 μ g/mL
Stock solution；The Standard Stock solutions for pipetting 1.0mL respectively are placed in same branch 100mL volumetric flask, are diluted to methanol/water
Scale shakes up, the level-one mixed standard solution that obtain concentration respectively be 1.0 μ g/mL；Accurately pipette 0.01,0.05,0.10,0.50,
1.00,5.00,10.0 and 50.0mL level-one mixed standard solution is respectively placed in different 100mL volumetric flasks, with compound chlorination
Sodium injection is diluted to scale, shakes up, and obtains equal 0.1,0.5,1.0,5.0,10.0,50.0, the 100.0 and 500.0ng/mL of concentration
Serial mixed standard solution.Serial mixed standard solution is subjected to high performance liquid chromatography-library by concentration sequence from low to high
The analysis of logical sequence array electrochemical detector, with the chromatographic peak areas of each target analytes (Y) to its corresponding concentration (X) carry out linearly
It returns, obtains standard curve regression equation.
E. data analysis and processing: the content of each object in dialysate samples is calculated according to calibration curve method, according to knot
Fruit draws the forward and backward each time point intracerebral monoamine neurotransmitter of fume exposure respectively and its Concentration-time variation of metabolin is bent
Line discloses the rule that fume exposure causes intracerebral monoamine neurotransmitter to change.
One aspect of the present invention utilizes animal mouth and nose formula fume exposure system, can be to be closer to mankind's smoking tobaccos system
The mode of product is exposed to animal in flue gas by way of only nasal inhalation, on the other hand also utilizes on-line microdialysis and efficient liquid
Phase chromatography-coulomb array electrochemical detector joint technology, can not only obtain the animal brain sample being exposed in flue gas in real time
And realize on-line checking monoamine neurotransmitter, while also there is high sensitivity, sampling interval time are short and analysis speed is fast etc.
Feature.5 kinds of monoamine neurotransmitter compounds in animal brain can not only be detected simultaneously using technical solution of the present invention, it is also real
Cause monoamine neurotransmitter changes in animal brain online, dynamic to measure when having showed fume exposure, takes compared with prior art
Obvious progress was obtained, a kind of new monitoring for Monoamine Metabolism variation in animal brain after fume exposure has been started
Method.
Detailed description of the invention
Fig. 1 is test mode schematic diagram of the present invention.
Serial number in Fig. 1: 1 be microdialysis infusion pump, 2 in clamper only by mouth and nose formula suck flue gas animal, 3
It is on-line automatic sample injector for microdialysis probe, 4,5 be high performance liquid chromatography-coulomb array electrochemical detector system).
Fig. 2 is the HPLC-ECD test map of 5 kinds of objects in rat encephalic micro-dialysis sample；
In figure: a. DA；b. 5-HT；c. DOPAC；d. 5-HIAA；e. HVA.
Fig. 3 is the Concentration-time trend chart of DA in the rat brain of cigarette smoke exposure front and back.
Fig. 4 is the Concentration-time trend chart of DOPAC in the rat brain of cigarette smoke exposure front and back.
Fig. 5 is the Concentration-time trend chart of HVA in the rat brain of cigarette smoke exposure front and back.
Fig. 6 is the Concentration-time trend chart of 5-HT in the rat brain of cigarette smoke exposure front and back.
Fig. 7 is the Concentration-time trend chart of 5-HIAA in the rat brain of cigarette smoke exposure front and back.
Fig. 8 is the Concentration-time trend chart of DA in the rat brain of electronic cigarette fume exposure front and back.
Fig. 9 is the Concentration-time trend chart of DOPAC in the rat brain of electronic cigarette fume exposure front and back.
Figure 10 is the Concentration-time trend chart of HVA in the rat brain of electronic cigarette fume exposure front and back.
Figure 11 is the Concentration-time trend chart of 5-HT in the rat brain of electronic cigarette fume exposure front and back.
Figure 12 is the Concentration-time trend chart of 5-HIAA in the rat brain of electronic cigarette fume exposure front and back.
Figure 13 is the dedicated experimental animal fixing clamp holder structural schematic diagram of the present invention.
In figure: 2-1 is front port, and 2-2 is that encephalic micro-dialysis samples skylight, and 2-3 is head lock chamber, and 2-4 is that body portion is fixed
Chamber, 2-5 are push rod, and 2-6 is end cap, and 2-7 is hold-doun nut, and 2-8 is that arc pushes head.
Specific embodiment
The present invention is described further with the following Examples, but is not intended to limit the present invention.
1. experimental material: male adult SD normal rat, 10 week old, weight (200 ± 20) g are raised using independent isolating
Cage tool raising is supported, is freely intake, standard particle feed.
2. major experimental equipment: animal mouth and nose suck fume exposure equipment, occur comprising rotating disc type smoking machine, aerosol
Device and animal mouth and nose sucking exposing device (HRH-MNE63150, the intelligent Rong He company in Beijing)；Microdialysis sgstem (including CMA/402
Microdialysis pump, the on-line automatic sample injector of CMA on-line injector, 12 microdialysis probe of CMA and conduit, Sweden CMA are public
Department)；SN-1N type stereotaxic apparatus (Japanese Narishige company)；High performance liquid chromatography-coulomb array electrochemical detector
(5600A CoulArray, Thermo company, the U.S.)；Improved experimental animal nose suck exposure holder structure referring to
Figure 13 (and separately applying for a patent).
3. experimental method: the method for the present invention includes the following steps:
(1) microdialysis probe conduit brain is implanted into: after rat weight, pressing 1.25 g/kg with 20% urethane ester solution
Dosage intraperitoneal injection of anesthesia, is fixed on stereotaxic instrument, cranium removal part endocranium is opened, according to positioning coordinate (AP:+2.5 mm；
ML:1.2 mm；DV:6.6 mm) spy micro dialysis needle casing is implanted into rat brain nucleus accumbens septi and is fixed with dental cement.Operation
Afterwards, rat single cage, which is fed, restores 24 hours.
(2) flue gas mouth and nose sucking exposure: rotating disc type smoking machine (each pumping volume under 3308 puffing regimens of ISO is utilized
It generates cigarette smoke for 35ml, suction 2s, interval 60s) smoking cigarette or tobacco juice for electronic smoke is added in aerosol generator
It generates electronic cigarette flue gas (aerosol), the experimental rat that brain is implanted with microdialysis probe conduit is placed in improved experimental animal
Nose sucks in exposure clamper, then accesses in animal mouth and nose sucking exposing device；Accurate control animal mouth and nose sucking exposure
The parameters such as temperature, the diluent air flow velocity of device are exposed to experimental rat in the flue gas of constant total granules (TPM) concentration.
(3) it microdialysis sampling: in the probe catheter of microdialysis probe (CMA 12) loading test rat head and will be connected to
Microdialysis pipeline uses the ringer's solution (compound sodium chloride injection) of filtration to sample with the speed perfusion of 2.0 μ L/min.
(4) microdialysis sampling pipeline sample detection: is passed through into on-line automatic sample injector and high performance liquid chromatography-coulomb array
Electrochemical detector connection with realize microdialysis sample it is on-line automatic sample introduction is analyzed, according to the chromatography row of 5 kinds of target compounds
For (Fig. 2) and analysis time (detection of an achievable sample in 14 min under following chromatographic conditions) setting it is on-line automatic into
The sampling interval of sample device is 15 min.
Chromatographic condition
Chromatographic column: Discovery HS F5 column (4.6 mm × 150 mm, 3 μm)；Column temperature: 30 DEG C；Mobile phase: A: first
Alcohol, B: buffering saline solution contains 40 mmol/L citric acids, 20 mmol/L disodium hydrogen phosphates and 0.3 mmol/L EDTA；Flow velocity:
0.25 mL/min；Condition of gradient elution is shown in Table 1；Sample volume: 20 μ L；According to the volt-ampere curve of each object, coulomb array is set
8 work potentials of electrochemical detector are as follows: 50,0,50,175,200,350,400,500 mV.
1 condition of gradient elution of table
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 10 | 90 |
| 4 | 25 | 75 |
| 13 | 25 | 75 |
| 14 | 10 | 90 |
(5) foundation of standard curve and sample data analysis
Serial mixed standard solution is subjected to HPLC- by concentration sequence from low to high in the case where not being connected to Microdialysis sgstem
ECD analysis, with the chromatographic peak area of each object (Y) to its corresponding concentration (X) linear regression is carried out, it obtains standard curve and returns
Return equation.
Quantitative analysis is carried out using 5 kinds of objects in the micro- sample of rat brain of the calibration curve method to detection, according to result
The Concentration-time change curve of the forward and backward each time point intracerebral monoamine neurotransmitter of fume exposure and its metabolin is drawn respectively,
Fume exposure is disclosed to the affecting laws of monoamine neurotransmitter.
Embodiment 1
Monoamine neurotransmitter variation detection test in rat brain after the exposure of certain reference cigarette smoke
Rat brain microdialysis probe conduit Implantation Test is carried out according to above-mentioned steps (1), is then carried out according to step (2)
Reference cigarette smoking and the sucking exposure of rat flue gas mouth and nose formula, make experimental rat be exposed to 200 mg/m of TPM concentration3Cigarette
In flue gas, microdialysis experiments then are carried out according to step (3), sample detection are carried out according to step (4), finally according to step (5)
Carry out the data analysis of sample.
Fig. 3, Fig. 4, Fig. 5, Fig. 6 and Fig. 7 be respectively cigarette smoke exposure front and back rat brain in DA, DOPAC, HVA, 5-HT and
The Concentration-time change curve schematic diagram of 5-HIAA.
The result shows that the release of DA and 5-HT continues to increase in rat nucleus accumbens septi, the primary of DA during cigarette smoke exposure
The content of metabolite DOPAC is without significant changes, and the content of DA and 5-HT respective whole grade metabolite HVA and 5-HIAA are bright
It is aobvious to increase, illustrate that the method for the present invention can be used for continuously monitoring the change of monoamine neurotransmitter in the rat brain of cigarette smoke exposure front and back
Law.
Embodiment 2
Monoamine neurotransmitter variation detection test in rat brain after certain electronic cigarette fume exposure
Rat brain microdialysis probe conduit Implantation Test is carried out according to above-mentioned steps (1), is then utilized according to step (2)
Aerosol generator carries out the generation and the sucking exposure of rat flue gas mouth and nose formula of electronics smoke sol, is exposed to experimental rat
TPM concentration is 1000 mg/m3Electronic cigarette flue gas in, then according to step (3) carry out microdialysis experiments, according to step (4) into
Row sample detection is finally analyzed according to the data that step (5) carry out sample.
Fig. 8, Fig. 9, Figure 10, Figure 11 and Figure 12 be respectively DA, DOPAC in rat brain before and after electronic cigarette fume exposure, HVA,
The Concentration-time change curve schematic diagram of 5-HT and 5-HIAA.The result shows that between electronic cigarette flue gas exposure period in rat nucleus accumbens septi
The variation of DA and 5-HT similar rule when showing with cigarette smoke exposure, illustrates that the method for the present invention can also be used for real-time monitoring
The change information of monoamine neurotransmitter in the rat brain of electronic cigarette fume exposure front and back.
Claims (3)
1. a kind of method that monoamine neurotransmitter changes in animal brain under the conditions of real-time detection fume exposure, it is characterised in that:
Animal is exposed in flue gas in real time using animal mouth and nose formula sucking fume exposure system, is collected by on-line microdialysis technology dynamic
Object brain tissue sample recycles high performance liquid chromatography-coulomb array electrochemical detector to the monoamine in brain dialysate samples
Neurotransmitter and its metabolin are quickly analyzed, thus monoamine neurotransmitter in the animal brain of real-time monitoring fume exposure front and back
Variation characteristic, the specific steps are as follows:
A. fume exposure: electronic cigarette liquid is made to generate aerosol using smoking machine smoking cigarette or using aerosol generator, by head
Portion is put into a dedicated experimental animal fixing clamp holder equipped with the animal of microdialysis probe conduit so that the clamper seize on both sides by the arms it is dynamic
Object is only exposed in certain density flue gas with mouth and nose suction；
B. encephalic micro-dialysis sample collection: will be continuous in the conduit of microdialysis probe insertion animal head after operation Microdialysis sgstem
Animal brain dialyzate is collected in the quantitative loop of online sample injector, operation is automatically injected by online sample injector at regular intervals
High performance liquid chromatography in；Chromatographic condition is specific as follows:
Chromatographic column: Discovery HS F5 column, specification 4.6 mm × 150 mm, 3 μm；Column temperature: 30 DEG C；Mobile phase: A: first
Alcohol, B: buffering saline solution contains 40 mmol/L citric acids, 20 mmol/L disodium hydrogen phosphates and 0.3 mmol/L EDTA；Flow velocity:
0.25 mL/min；The multiple channel operation potential of coulomb array electrochemical detector is set according to the volt-ampere curve of each object
Are as follows: 50,0,50,175,200,350,400,500 mV；Condition of gradient elution is as follows:
C. high performance liquid chromatography-coulomb array electrochemical detector analysis: setting gradient elution program and sample introduction sequence are simultaneously run
High performance liquid chromatography-coulomb array electrochemical detector detects microdialysis sample, so that the detection time of each sample is slightly less than
The acquisition time of the sample；
D. standard solution prepare and Specification Curve of Increasing: prepare containing dopamine (DA), 3,4 dihydroxyphenyl acetic acids (DOPAC),
Homovanillic acid (HVA), the serial hybrid standard of 5 kinds of standard items of serotonin (5-HT) and 5 hydroxyindole acetic acids (5-HIAA) are molten
Serial mixed standard solution is carried out high performance liquid chromatography-coulomb array electrochemical by concentration sequence from low to high and detected by liquid
Device analysis, with the chromatographic peak areas of each target analytes (Y) to its corresponding concentration (X) linear regression is carried out, obtain standard song
Line regression equation；
E. data analysis and processing: the content of each object in dialysate samples is calculated according to calibration curve method, according to result point
Not Hui Zhi the forward and backward each time point intracerebral monoamine neurotransmitter of fume exposure and its metabolin Concentration-time change curve, take off
Show the rule that fume exposure causes intracerebral monoamine neurotransmitter to change.
2. the side that monoamine neurotransmitter changes in animal brain under the conditions of real-time detection fume exposure according to claim 1
Method, it is characterised in that: standard solution process for preparation is specific as follows: 5 kinds of DA, DOPAC, HVA, 5-HT and 5-HIAA are weighed respectively
Each 10 mg of standard items, is respectively placed in 100 mL volumetric flasks, is dissolved and is diluted to the methanol/water solution that volume ratio is 20:80
Scale shakes up, the Standard Stock solutions that obtain concentration respectively be 100 μ g/mL；The Standard Stock solutions for pipetting 1.0 mL respectively, are placed in
In same branch 100mL volumetric flask, it is diluted to scale with the methanol/water solution that volume ratio is 20:80, is shaken up, obtaining concentration is respectively 1.0
The level-one mixed standard solution of μ g/mL；Accurately pipette 0.01,0.05,0.10,0.50,1.00,5.00,10.0 and 50.0mL mono-
Grade mixed standard solution, is respectively placed in different 100mL volumetric flasks, is diluted to scale with compound sodium chloride injection, shakes up,
Obtain the serial mixed standard solution of equal 0.1,0.5,1.0,5.0,10.0,50.0,100.0 and 500.0 ng/mL of concentration.
3. the side that monoamine neurotransmitter changes in animal brain under the conditions of real-time detection fume exposure according to claim 1
Method, it is characterised in that: the animal is rat.
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| CN108459095A (en) * | 2018-01-17 | 2018-08-28 | 烟台大学 | Detection method that is a kind of while detecting eight kinds of neurotransmitters |
| CN108802250B (en) * | 2018-08-28 | 2020-09-25 | 上海中医药大学 | Method for detecting 11 neurotransmitters in brain microdialysis solution by ultra-high performance liquid chromatography-mass spectrometry |
| CN112730738A (en) * | 2020-12-07 | 2021-04-30 | 中国科学院深圳先进技术研究院 | Evaluation device and evaluation method for reward and addiction of atomized matter |
| CN114609300A (en) * | 2022-04-07 | 2022-06-10 | 国家烟草质量监督检验中心 | Method for evaluating influence of menthol addition on nicotine metabolism of oral nicotine based on animal model |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102455313A (en) * | 2010-11-01 | 2012-05-16 | 上海纳米技术及应用国家工程研究中心有限公司 | Lamina detection pool used for detecting dopamine and preparation method thereof |
| CN104020241A (en) * | 2014-06-11 | 2014-09-03 | 中国烟草总公司郑州烟草研究院 | Synchronous analysis method for trace nicotine in blood-brain samples of animal and main metabolites thereof |
| RU2013113102A (en) * | 2013-03-25 | 2014-09-27 | Татьяна Михайловна Рожнова | METHOD FOR FORECASTING RISK OF DEVELOPMENT OF DISORDERS OF BEHAVIOR OF A DEPENDENT CHARACTER IN THE FORM OF ADDICTION, ALCOHOLISM, TOBACCO SMOKING AND / OR THE ACCEPTANCE PHENOMENON |
| CN105891357A (en) * | 2016-04-05 | 2016-08-24 | 中国烟草总公司郑州烟草研究院 | Analysis method for dynamically tracing and monitoring nicotine and nicotine metabolites in animal |
-
2016
- 2016-12-29 CN CN201611241605.5A patent/CN106645492B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102455313A (en) * | 2010-11-01 | 2012-05-16 | 上海纳米技术及应用国家工程研究中心有限公司 | Lamina detection pool used for detecting dopamine and preparation method thereof |
| RU2013113102A (en) * | 2013-03-25 | 2014-09-27 | Татьяна Михайловна Рожнова | METHOD FOR FORECASTING RISK OF DEVELOPMENT OF DISORDERS OF BEHAVIOR OF A DEPENDENT CHARACTER IN THE FORM OF ADDICTION, ALCOHOLISM, TOBACCO SMOKING AND / OR THE ACCEPTANCE PHENOMENON |
| CN104020241A (en) * | 2014-06-11 | 2014-09-03 | 中国烟草总公司郑州烟草研究院 | Synchronous analysis method for trace nicotine in blood-brain samples of animal and main metabolites thereof |
| CN105891357A (en) * | 2016-04-05 | 2016-08-24 | 中国烟草总公司郑州烟草研究院 | Analysis method for dynamically tracing and monitoring nicotine and nicotine metabolites in animal |
Non-Patent Citations (3)
| Title |
|---|
| Effect of venlafaxine and nicotine on the level of neurotransmitters and their metabolites in rat brains;Anna Czubak等;《Journal of physiology and Pharmacology》;20101231;第61卷(第3期);339-346 |
| 不同模式口鼻式吸烟诱导急性肺损伤的实验研究;肖卫强等;《浙江大学学报（医学版）》;20160930;第45卷(第5期);522-529 |
| 库仑阵列电化学-高效液相色谱法同时检测大鼠不同脑区中的8种单胺类神经递质;邓少东等;《广东医学院学报》;20151231;第33卷(第6期);644-648 |
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