CN103808846B - Series quadrupole-rod gas-chromatographic mass spectrometry detection method for 35 toxic medicaments in urine - Google Patents
Series quadrupole-rod gas-chromatographic mass spectrometry detection method for 35 toxic medicaments in urine Download PDFInfo
- Publication number
- CN103808846B CN103808846B CN201410057537.1A CN201410057537A CN103808846B CN 103808846 B CN103808846 B CN 103808846B CN 201410057537 A CN201410057537 A CN 201410057537A CN 103808846 B CN103808846 B CN 103808846B
- Authority
- CN
- China
- Prior art keywords
- urine
- kinds
- drugs
- temperature
- ion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a series quadrupole-rod gas-chromatographic mass spectrometry detection method for 35 toxic medicaments in urine. The series quadrupole-rod gas-chromatographic mass spectrometry detection method comprises the steps of constructing a multiple-reaction detection method on a series quadrupole-rod gas chromatograph-mass spectrometer through standard substances of 35 toxicants (medicines), and determining 2-3 pairs of feature parent ions and daughter ions and the retention time of each toxicants (medicines); extracting a urine sample to be detected through diethyl ether, performing ultrasonic and centrifugal treatment, blow-drying and dissolving, and then qualitatively and quantitatively analyzing the toxicants (medicines) through a tandem mass spectrum MRM. By the adoption of a series quadrupole-rod gas-chromatographic-mass spectrometry/mass spectrometry method (GC-MS/MS), the method disclosed by the invention can quickly screen 35 conventional toxicants (medicines) in the urine of a human body; a detection result is accurate, sensitive and quick; the recycling rate of toxicants (medicines) is 80.82-118.56 percent; the detection limit is up to 0.0014-1.8 micrograms per microliter.
Description
Technical field
The invention belongs to the series connection level Four bar gaschromatographic mass spectrometry detection method of 35 kinds of poisonous substances in a kind of urine.
Background technology
Drug abuse, traffic in drugs have become global social effects of pollution.People once be infected with drugs will often wallow in drugs among the deformity " pleasant sensation " had fantasies of out, he is not in the mood for working and learning.Take drugs and also can cause a series of family social problem.In view of consume illegal drugs is to family, particularly to the serious harm of society, abroad some countries are except taking corresponding control measures to national drug addict, immigrant also required to the proof providing applicant not take drugs, as Russia, Belgium, Hong Kong etc.Along with global economic integration progress is accelerated, other countries inevitably will put into effect similar regulation successively to immigrant.Propose to add the requirement doing illicit drugs inspection to relating to health status aspect during my personnel's entry visa for responding actively more external countries, inspection and quarantine department will according to the requirement of State Bureau, actively carries out material and technical preparation on the one hand to tackle the external requirement to my entry personnel's health examination; And maintain social stability object healthy from protection our people, also should take the principle of equity, the personnel of coming to China be increased to the requirement of illicit drugs inspection on the other hand.
In the most widely used domestic and international human body, the measuring technology of drug ingredient inspection is immunoassay techniques and gas chromatography (GC) at present, abroad also has (GC/MS) that use gas chromatography mass spectrometry to detect.Retrieval Chinese Industrial Standards (CIS) net finds, the professional standard about a few class drugs such as morphine, cocaine, heroin of the Jin You Ministry of Public Security in a standard, the mainly direct detection to drugs, and is not the detection method for clinical sample aspect.Current inspection and quarantine department mainly adopts colloidal gold technique to detect morphine and meth fast to the method that inward and outward personnel's urine carries out illicit drugs inspection, and false positive rate is high, and can only detect 2 kinds of drugs.Therefore set up a kind of quick, sensitive, accurately multiple poison (medicine) thing Simultaneous Detection become particularly important.
China Patent No. is the 201110075177.4 Liquid Chromatography-Tandem Mass Spectrometry detection methods disclosing 33 kinds of drugs in a kind of urine, and its step is as follows:
Sample treatment: take 2g urine sample to be checked, be accurate to 0.01g, be placed in 50mL tool plug centrifuge tube, add 10ml ethyl acetate, liquid blending device fully mixes 1min, with the centrifugal 10min of 4000r/min, draw upper strata ethyl acetate in test tube, then add 1mL0.1mol/L NaOH solution, after liquid blending device fully mixes, add 10ml ethyl acetate to repeat to extract once, merge extracted twice liquid; Extract nitrogen dries up instrument and dries up at 40 DEG C, adds the methyl alcohol that 1mL volume ratio is 30: 70: eddies of water supination, crosses 0.25 μm of filter membrane, obtains testing sample;
Measure: testing sample liquid chromatography tandem quadrupole mass spectrometer measures; In the Liquid Chromatography-Tandem Mass Spectrometry analytic approach of 33 kinds of drugs, except barbiturate uses negative ion mode analysis, all the other compounds all use positive ion mode analysis;
Wherein the Mass Spectrometry Conditions of positive ion mode is: ionization source: ESI source; Scan pattern: cation selects scan pattern SRM; Electron spray voltage: 2.5kV; Dry gas temperature: 400 DEG C; Gas curtain atmospheric pressure: 45arb; Assisted gas pressure: 7arb; Capillary temperature: 350 DEG C; Collision gas: high-purity argon gas;
The Mass Spectrometry Conditions of negative ion mode is: ionization source: ESI source; Scan pattern: anion selects scan pattern SRM; Electron spray voltage: 2.5kV; Dry gas temperature: 400 DEG C; Gas curtain atmospheric pressure: 50arb; Assisted gas pressure: 7arb; Capillary temperature: 320 DEG C; Collision gas: high-purity argon gas;
Chromatographic condition is: chromatographic column: Phenomenex Gemini C18100 × 2.0,3 μm; Column temperature: room temperature; Flow velocity: 250 μ L/min; Sampling volume: 25 μ L.
China Patent No. is that the liquid-liquid extraction method of its urine of documents employing classics of 201110075177.4 carries out sample pre-treatments, and adopt ethyl acetate as extractant, extractant shares amount 20ml, and can only detect 33 kinds of drugs.This method and adopt Liquid Chromatography-Tandem Mass Spectrometry detect, the weak point of its method is, due to detection material some with cation scanning and some for anion scan.Real disposable detection can not be reached so will be divided into when detecting carrying out for twice detecting.
Summary of the invention
The object of this invention is to provide and a kind of adopt series connection level Four bar combined gas chromatography mass spectrometry (GC-MS/MS) new method of common poison (medicine) thing in 35 in Rapid Screening human urine simultaneously.
The object of the present invention is achieved like this, the series connection level Four bar gas chromatography combined with mass spectrometry detection method of 35 kinds of poison (medicine) things in described urine, it is characterized in that: first use the standard items of 35 kinds of poison (medicine) to set up multiple reaction detection method on series connection level Four bar GC-MS, determine 2 ~ 3 pairs of feature parent ions and daughter ion to and the retention time of often kind of poison (medicine) thing; Urine sample extracted with diethyl ether to be measured, through ultrasonic, centrifugal, dry up and dissolve after, with tandem mass spectrum MRM qualitative and quantitative analysis poison (medicine) thing;
Chromatographic condition is: instrument, Agilent7890GC/7000QQB; GC condition: DB-1MS(30m × 0.25mm × 0.25 μm) quartz capillary column, injector temperature: 250 DEG C, do not shunt; Temperature programming: initial temperature 50 DEG C, is warmed up to 290 DEG C with 12 DEG C/min and keeps 10min, flow velocity: 1ml/min, sample size: 0.5 μ L; MS condition: EI ion gun, electronic energy 70eV, ion source temperature 230 DEG C, interface temperature: 280 DEG C, level Four bar temperature: 150 DEG C, electron multiplier voltage: 1381eV.
Described series connection level Four bar gas chromatography combined with mass spectrometry detection method, specifically comprise: 1) standard liquid preparation: add DDVP in the methanol solution of 35 kinds of poison (medicine) thing hybrid standard product, Rogor, parathion-methyl, malathion, parathion, Tetramine concentration are 22.73 μ g/ml; Barbital, amytal, quinalbarbitone, lidocaine, phenobarbital, atropine, amitriptyline, chlorpromazine, midazolam, nitrazepam, Clonazepam, Clozapine, estazolam, alprazolam, triazolam concentration are 16.67 μ g/ml; MDMA, paracetamol, pethidine, aminopyrine, C16H25NO2, chlorpheniramine, methadone, doxepin, carbamazepine, diazepam concentration are 20.83 μ g/ml;
Crystal methamphetamine, ephedrine, ketamine, codeine concentration are 25 μ g/ml;
2) detection method
2.1) sample treatment
Get urine 1ml to be checked, add ether 6ml and divide three extractions, each 2ml; At 40 DEG C of ultrasonic extraction 10min, then with the centrifugal 3min of 10000r/min, the merging of three extraction organic layers is transferred in another test tube, and at 40 DEG C, nitrogen dries up, and residue 100 μ L methyl alcohol dissolve, and gets 1 μ L and enters GC/MS/MS analysis;
2.2) GC condition
GC condition: DB-1 (30m × 0.25mm × 0.25m) quartz capillary column, injector temperature: 250 DEG C, do not shunt; Temperature programming: initial temperature 50 DEG C, is warmed up to 290 DEG C with 12 DEG C/min and keeps 10min, flow velocity: 1ml/min, sample size: 0.5 μ L;
2.3) MS condition
MS condition: EI ion gun, electronic energy 70eV, ion source temperature 230 DEG C, interface temperature: 280 DEG C, level Four bar temperature: 150 DEG C, electron multiplier voltage: 1381eV;
The optimum parent ion daughter ion of 35 kinds of cytotoxic drugs is to seeing the following form 1:
Table 135 kind of poison (medicine) thing chromatographic retention and mass spectral characteristic ion pair
* in table is quota ion pair;
3) qualitative-and-quantitative method: be qualitative foundation with the abundance ratio that the monitoring parent ion/daughter ion of the relative retention time of each drugs and each drugs is right, the retention time of each drugs sees the above table 1; With quantified by external standard method, with its residual quantity in the sample to which of the calculated by peak area of each drugs;
4) matrix effect
After blank urine sample is by above-mentioned " 2.1 " step process, adds the standard liquid of 35 kinds of poison (medicine) things, carry out GC-MS/MS analysis, obtain standard items peak area A; Equally the standard liquid of 35 kinds of poison (medicine) things is analyzed, obtain corresponding peak area B; Matrix effect=A/B × 100%, result shows that except nitrazepam and Clonazepam are other 33 kinds of cytotoxic drugs of substrate inhibition be all matrix enhancement, and wherein atropinic matrix enhancement effect is the most obvious, specifically sees the following form 2;
Table 235 kind of cytotoxic drug matrix effect list
5) calibration curve and detectability
Get blank urine sample, add 35 kinds of poison (medicine) thing standard liquid mixing, be made into paracetamol mass concentration in urine and be respectively 3.0,4.0,5.0,7.5,10.0,15.0 μ g/mL; In urine, DDVP mass concentration is respectively 2.0,3.0,4.0,5.0,7.5,10.0 μ g/mL; In urine, Tetramine, Rogor, parathion-methyl, malathion and parathion mass concentration are respectively 2.0,3.0,4.0,5.0,7.5,9.1 μ g/mL; In urine, crystal methamphetamine, ephedrine, ketamine and codeine mass concentration are respectively 0.2,0.5,1.0,2.5,5.0,7.5 μ g/mL; In urine, all the other 24 kinds of cytotoxic drug mass concentrations are respectively the standard liquid of 2.0,3.0,4.0,4.5,5.0,6.5 μ g/mL; GC-MS/MS analysis is carried out by after 2.1 step method pre-treatments; With cytotoxic drug peak area (A) to cytotoxic drug mass concentration (C, μ g/mL) drawing standard curve, and try to achieve linear regression equation and coefficient correlation; The urine intoxicant substrate concentration (with 3 times signal-to-noise ratio computation) of minimum detectability for detecting by 2.1 step method processing samples, in the visible the method for result, the linear equation correlation coefficient r of 35 kinds of cytotoxic drugs is more than 0.98, specifically in table 3;
The calibration curve equation of table 335 kind of poison (medicine) thing, coefficient correlation and detection limit
The main technical content of detection method of the present invention and general principle:
1) extractant contrast cyclohexane, ethyl acetate, chloroform and ether four kinds, find that extracted by ether rate is high, and ether boiling point is low, more easily concentrates, and therefore selects ether as extractant.
2) the present invention is by groping and the optimization of GC-MS/MS analysis condition sample-pretreating method, by extracted with diethyl ether, segmentation multiple reaction monitoring method (MRM) is identified, and verify further by extraction ion pair, the 7 large classes such as acephatemet, Tetramine, diazepam, nikethamidum, lidocaine, phenobarbital, atropine common poison (medicine) thing in totally 35 can be detected simultaneously, the rate of recovery reaches 80.82% to 118.56%, and detection limit reaches 0.0014 ~ 1.8 μ g/ml.This law chromatographic retention, mass spectral characteristic parent ion/daughter ion to simultaneously qualitative, with its residual quantity in the sample to which of the calculated by peak area of each cytotoxic drug.The method eliminates the interference of complicated substrate in urine to greatest extent, quick and precisely can carry out qualitative and quantitative analysis to 35 kinds of poison (medicine) things, widely applicable, simple to operate, detect with strong points.Can be used for inspection and quarantining for import/export, disease prevention and control center, public security department is detected and the confirmation of positive findings fast.
With National Standard Method contrast, this method has the features such as sensitive, accurate, easy, quick.
Advantage of the present invention is: 1) in urine, the routine techniques pre-treatment of common cytotoxic drug adopts liquid-liquid extraction method, and the present invention finds that extracted by ether rate is high, shares 6mL ether and just can extract completely 35 poisoning (medicine) thing in urine.And ether boiling point is low, more easily concentrates, therefore select ether as extractant; 2) documents of routine techniques if the patent No. is 201110075177.4 adopts high performance liquid chromatography-tandem mass technology, it can only detect 33 kinds of poison (medicine) things, the present invention adopts series connection level Four bar combined gas chromatography mass spectrometry (GC-MS/MS) simultaneously common poison (medicine) thing in 35 in Rapid Screening human urine, assay is accurate, sensitive, quick, poison (medicine) thing rate of recovery reaches 80.82% to 118.56%, and detection limit reaches 0.0014 ~ 1.8 μ g/ml.
Accompanying drawing explanation
Fig. 1 is 35 kinds of cytotoxic drug standard sample GC-MS/MS multiple-reaction monitoring spectrograms.As can be seen from FIG. 35 kinds of cytotoxic drugs retention time and go out peak position.
Detailed description of the invention
Below in conjunction with example, the present invention is described in detail
1 instrument
Instrument: Agilent7890GC/7000QQQB.
2 reagent
2.1 medicines: in the standard items of 35 kinds of poison (medicine) things in measurement range, diazepam and MDMA are the methanol solution of 500 μ g/ml, and Tetramine is 250 μ g/ml ethyl acetate solutions, and other is 1mg/mL methanol solution.Purchased from Expert Testimony Science-Technology Inst., Judical Department.
2.2 reagent: cyclohexane, ethyl acetate, it is pure that chloroform, ether, phosphoric acid, sodium dihydrogen phosphate, NaOH, sodium carbonate, sodium acid carbonate, hydrochloric acid etc. are analysis; Methyl alcohol is chromatographically pure.
2.3 standard liquids preparation: add DDVP in the methanol solution of 35 kinds of poison (medicine) thing hybrid standard product, Rogor, parathion-methyl, malathion, parathion, Tetramine concentration are 22.73 μ g/ml; Barbital, amytal, quinalbarbitone, lidocaine, phenobarbital, atropine, amitriptyline, chlorpromazine, midazolam, nitrazepam, Clonazepam, Clozapine, estazolam, alprazolam, triazolam concentration are 16.67 μ g/ml; MDMA, paracetamol, pethidine, aminopyrine, C16H25NO2, chlorpheniramine, methadone, doxepin, carbamazepine, diazepam concentration are 20.83 μ g/ml; Crystal methamphetamine, ephedrine, ketamine, codeine concentration are 25 μ g/ml.
3 analytical methods
3.1 sample treatment
Get urine 1ml to be checked, add ether 6ml and divide three extractions, each 2ml.At 40 DEG C of ultrasonic extraction 10min, then with the centrifugal 3min of 10000r/min, the merging of three extraction organic layers is transferred in another test tube, and at 40 DEG C, nitrogen dries up, and residue 100 μ L methyl alcohol dissolve, and gets 1 μ L and enters GC/MS/MS analysis.
3.2GC condition
GC condition: DB-1 (30m × 0.25mm × 0.25 μm) quartz capillary column, injector temperature: 250 DEG C, do not shunt.Temperature programming: initial temperature 50 DEG C, is warmed up to 290 DEG C with 12 DEG C/min and keeps 10min, flow velocity: 1ml/min, sample size: 0.5 μ L.
3.3MS condition
MS condition: EI ion gun, electronic energy 70eV, ion source temperature 230 DEG C, interface temperature: 280 DEG C, level Four bar temperature: 150 DEG C, electron multiplier voltage: 1381eV.
4 method optimization and method validations
4.1 sample pre-treatments optimizations
4.1.1 the optimization of extractant
Extractant contrast cyclohexane, ethyl acetate, chloroform and ether four kinds, find that extracted by ether rate is high, and ether boiling point is low, more easily concentrates, and therefore selects ether as extractant.
4.1.2 the optimization of pH value is extracted
The pH value of regulation system is respectively 1.5,3.5,5,7,9,12.Result is according to the peak area of each poison (medicine) thing, and the pH when rate of recovery is totally high is 7.
The optimization of 4.2GC condition
Contrast chromatographic column DB-1MS, HP-5MS, DB-17MS.BD-1MS can make 35 kinds of poison (medicine) thing compositions be well separated.
The optimization of 4.3MS condition
4.3.1 the feature parent ion daughter ion pair of compound to be checked is determined
First use the standard items of 35 kinds of poison (medicine) things on GC/MS/MS, set up multiple reaction monitoring (MRM) method, determine feature three pairs of parent ions and the daughter ion pair of compound to be checked.
4.3.2 the exploitation of sectional detecting method
The hybrid standard product of 35 kinds of poison (medicine) things are detected by this MRM method, suitable optimization chromatographic condition makes 35 kinds of poison (medicine) things substantially can reach baseline separation respectively, therefore adopt each material to be divided into an analysis period because each thing to be checked can reach baseline separation substantially, select the feature MRM method going out the cytotoxic drug composition at peak at this section to detect.Final retention time and feature parent ion and daughter ion are to qualitative.When two materials are when same retention time goes out peak or appearance time difference is very little, can adopt and extract parent ion daughter ion the qualitative and Quantitative Separation analysis of compound is carried out to chromatogram.
5 interpretations of result
The optimum parent ion daughter ion of 5.135 kinds of cytotoxic drugs is to being listed as follows:
Table 135 kind of poison (medicine) thing chromatographic retention and mass spectral characteristic ion pair
Note: * is quota ion pair.
The spectrogram of 5.235 kinds of cytotoxic drugs when optimum MRM method and sectional detecting method is shown in Fig. 1, and Figure 135 plants cytotoxic drug standard sample GC-MS/MS multiple-reaction monitoring spectrogram, as can be seen from FIG. 35 kinds of cytotoxic drugs retention time and go out peak position.5.3 qualitative-and-quantitative methods: be qualitative foundation with the abundance ratio that the monitoring parent ion/daughter ion of the relative retention time of each drugs and each drugs is right, the retention time of each drugs sees the above table; With quantified by external standard method, with its residual quantity in the sample to which of the calculated by peak area of each drugs.
5.4 specificity
Due to the advance of GC-MS/MS method, adopting the feature parent ion daughter ion of target analytes to carrying out qualitative and quantitative analysis, farthest can get rid of matrix interference and improving the specificity that object is detected.
5.5 matrix effect
In blank urine sample by after " 3.1 " item process, add the standard liquid of 35 kinds of poison (medicine) things, carry out GC-MS/MS analysis, obtain standard items peak area A; Equally the standard liquid of 35 kinds of poison (medicine) things is analyzed, obtain corresponding peak area B; Matrix effect=A/B × 100%.Result shows that except nitrazepam and Clonazepam are other 33 kinds of cytotoxic drugs of substrate inhibition be all matrix enhancement, and wherein atropinic matrix enhancement effect is the most obvious, specifically sees the following form 2.
Table 235 kind of cytotoxic drug matrix effect list
5.6 calibration curves and detectability
Get blank urine sample, add 35 kinds of poison (medicine) thing standard liquid mixing, be made into paracetamol mass concentration in urine and be respectively 3.0,4.0,5.0,7.5,10.0,15.0 μ g/mL; In urine, DDVP mass concentration is respectively 2.0,3.0,4.0,5.0,7.5,10.0 μ g/mL; In urine, Tetramine, Rogor, parathion-methyl, malathion and parathion mass concentration are respectively 2.0,3.0,4.0,5.0,7.5,9.1 μ g/mL; In urine, crystal methamphetamine, ephedrine, ketamine and codeine mass concentration are respectively 0.2,0.5,1.0,2.5,5.0,7.5 μ g/mL; In urine, all the other 24 kinds of cytotoxic drug mass concentrations are respectively the standard liquid of 2.0,3.0,4.0,4.5,5.0,6.5 μ g/mL.GC-MS/MS analysis is carried out by after 3.1 method pre-treatments.With cytotoxic drug peak area (A) to cytotoxic drug mass concentration (C, μ g/mL) drawing standard curve, and try to achieve linear regression equation and coefficient correlation.The urine intoxicant substrate concentration (with 3 times signal-to-noise ratio computation) of minimum detectability for detecting by 3.1 method processing samples, in the visible the method for result, the linear equation correlation coefficient r of 35 kinds of cytotoxic drugs is more than 0.98, specifically in table 3.
The calibration curve equation of table 335 kind of poison (medicine) thing, coefficient correlation and detection limit
5.7 degrees of accuracy and precision
Operate by under step " preparations of 4.5 calibration curves " item, prepare quality control (QC) sample of 35 kinds of basic, normal, high three concentration of poison (medicine) thing, each concentration is carried out 5 times and is analyzed, METHOD FOR CONTINUOUS DETERMINATION five days, according to the calibration curve on the same day, what calculate QC sample records concentration, calculates the veracity and precision of this law, the results are shown in Table 4 according to QC sample result.
In a few days and in the daytime the rate of recovery and the precision of table 435 kind of poison (medicine) thing
6 full patterns measure
Get actual sample, wherein known to DDVP, Tetramine, amytal, ketamine, doxepin and Clozapine 6 kinds of poison (medicine) things, and the concentration of known six kinds of poisonous substances is 5 μ g/mL.This actual sample processes according to step " 3.1 " sample-pretreating method, and carry out GC-MS/MS analysis, measurement result is in table 5.As known from Table 5, adopt 6 kinds of poison (medicine) things in detection method actual sample of the present invention all accurately qualitative, the relative deviation of the content obtained by quantitation curves and actual sample amount is all less than 5%, proves that this detection method is reliable and practical.
In table 5 people urine, actual sample measures the result table of poison (medicine) thing content
7 discuss
The present invention utilizes GC-MS/MS to have the advantages such as very high specificity, sensitivity, accuracy, establish the disposable new method simultaneously detecting 35 kinds of common cytotoxic drugs, system thinking and optimization are carried out to the sample pre-treatments such as Extraction solvent, system pH, chromatographic column and testing conditions, has determined the optimum parent ion/daughter ion of 35 kinds of poison to (specifically in table 1).Under selected conditions, furthermore present the detectability (specifically in table 3) etc. of the range of linearity of 35 kinds of cytotoxic drugs on GC-MS/MS, linear equation (all linearly dependent coefficient r are all greater than 0.98), often kind of material, 35 kinds of cytotoxic drug institute method for building up in a few days and in the daytime the rate of recovery is all between 80% ~ 120%, precision RSD value is all less than 10%.
The present invention uses GC-MS/MS method in a word, disposablely can carry out qualitative and quantitative analysis to 35 kinds of cytotoxic drugs simultaneously, with qualitative while of chromatographic retention and mass spectrum, eliminate other composition interference in urine, result accurately and reliably, selective and reproducible, actual sample qualitative and quantitative detection all obtains satisfactory result, shows that the GC-MS/MS method set up can meet the requirement of illicit drugs inspection in human urine accurately and reliably.
Claims (1)
1. the series connection level Four bar gas chromatography combined with mass spectrometry detection method of 35 kinds of cytotoxic drugs in a urine, it is characterized in that: first on series connection level Four bar GC-MS, set up multiple reaction detection method with the standard items of 35 kinds of poison, determine 2 ~ 3 pairs of feature parent ions and daughter ion to and the retention time of often kind of cytotoxic drug; Urine sample extracted with diethyl ether to be measured, through ultrasonic, centrifugal, dry up and dissolve after, with tandem mass spectrum MRM qualitative and quantitative analysis cytotoxic drug;
Chromatographic condition is: instrument, Agilent 7890GC/7000QQB; GC condition: DB-1MS 30m × 0.25mm × 0.25 μm quartz capillary column, injector temperature: 250 DEG C, do not shunt; Temperature programming: initial temperature 50 DEG C, is warmed up to 290 DEG C with 12 DEG C/min and keeps 10min, flow velocity: 1ml/min, sample size: 0.5 μ L; MS condition: EI ion gun, electronic energy 70eV, ion source temperature 230 DEG C, interface temperature: 280 DEG C, level Four bar temperature: 150 DEG C, electron multiplier voltage: 1381eV;
Concrete detecting step is:
1) standard liquid preparation: add DDVP in the methanol solution of 35 kinds of cytotoxic drug hybrid standard product, Rogor, parathion-methyl, malathion, parathion, Tetramine concentration are 22.73 μ g/ml; Barbital, amytal, quinalbarbitone, lidocaine, phenobarbital, atropine, amitriptyline, chlorpromazine, midazolam, nitrazepam, Clonazepam, Clozapine, estazolam, alprazolam, triazolam concentration are 16.67 μ g/ml; MDMA, paracetamol, pethidine, aminopyrine, C16H25NO2, chlorpheniramine, methadone, doxepin, carbamazepine, diazepam concentration are 20.83 μ g/ml; Crystal methamphetamine, ephedrine, ketamine, codeine concentration are 25 μ g/ml;
2) detection method
2.1) sample treatment
Get urine 1ml to be checked, add ether 6ml and divide three extractions, each 2ml; At 40 DEG C of ultrasonic extraction 10min, then with the centrifugal 3min of 10000r/min, the merging of three extraction organic layers is transferred in another test tube, and at 40 DEG C, nitrogen dries up, and residue 100 μ L methyl alcohol dissolve, and gets 1 μ L and enters GC/MS/MS analysis;
2.2) GC condition
GC condition: DB-130m × 0.25mm × 0.25m quartz capillary column, injector temperature: 250 DEG C, do not shunt; Temperature programming: initial temperature 50 DEG C, is warmed up to 290 DEG C with 12 DEG C/min and keeps 10min, flow velocity: 1ml/min, sample size: 0.5 μ L;
2.3) MS condition
MS condition: EI ion gun, electronic energy 70eV, ion source temperature 230 DEG C, interface temperature: 280 DEG C, level Four bar temperature: 150 DEG C, electron multiplier voltage: 1381eV;
The optimum parent ion daughter ion of 35 kinds of cytotoxic drugs is to seeing the following form 1:
Table 1 35 kinds of cytotoxic drug chromatographic retentions and mass spectral characteristic ion pair
* in upper table is quota ion pair;
3) qualitative-and-quantitative method: be qualitative foundation with the abundance ratio that the monitoring parent ion/daughter ion of the relative retention time of each drugs and each drugs is right, the retention time of each drugs sees the above table 1; With quantified by external standard method, with its residual quantity in the sample to which of the calculated by peak area of each drugs;
4) matrix effect
In blank urine sample by above-mentioned 2.1) after step process, add the standard liquid of 35 kinds of cytotoxic drugs, carry out GC-MS/MS analysis, obtain standard items peak area A; Equally the standard liquid of 35 kinds of cytotoxic drugs is analyzed, obtain corresponding peak area B; Matrix effect=A/B × 100%, result shows that except nitrazepam and Clonazepam are other 33 kinds of cytotoxic drugs of substrate inhibition be all matrix enhancement, and wherein atropinic matrix enhancement effect is the most obvious, specifically sees the following form 2;
The list of table 2 35 kinds of cytotoxic drug matrix effects
5) calibration curve and detectability
Get blank urine sample, add 35 kinds of cytotoxic drug standard liquid mixing, be made into paracetamol mass concentration in urine and be respectively 3.0,4.0,5.0,7.5,10.0,15.0 μ g/mL; In urine, DDVP mass concentration is respectively 2.0,3.0,4.0,5.0,7.5,10.0 μ g/mL; In urine, Tetramine, Rogor, parathion-methyl, malathion and parathion mass concentration are respectively 2.0,3.0,4.0,5.0,7.5,9.1 μ g/mL; In urine, crystal methamphetamine, ephedrine, ketamine and codeine mass concentration are respectively 0.2,0.5,1.0,2.5,5.0,7.5 μ g/mL; In urine, all the other 24 kinds of cytotoxic drug mass concentrations are respectively the standard liquid of 2.0,3.0,4.0,4.5,5.0,6.5 μ g/mL; GC-MS/MS analysis is carried out by after 2.1 step method pre-treatments; With cytotoxic drug peak area A to cytotoxic drug mass concentration C, μ g/mL drawing standard curve, and try to achieve linear regression equation and coefficient correlation; Minimum detectability is for the urine intoxicant substrate concentration that can detect by 2.1 step method processing samples is with 3 times of signal-to-noise ratio computation, and in the visible the method for result, the linear equation correlation coefficient r of 35 kinds of cytotoxic drugs is more than 0.98, specifically in table 3;
The calibration curve equation of table 3 35 kinds of cytotoxic drugs, coefficient correlation and detection limit
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410057537.1A CN103808846B (en) | 2014-02-20 | 2014-02-20 | Series quadrupole-rod gas-chromatographic mass spectrometry detection method for 35 toxic medicaments in urine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410057537.1A CN103808846B (en) | 2014-02-20 | 2014-02-20 | Series quadrupole-rod gas-chromatographic mass spectrometry detection method for 35 toxic medicaments in urine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103808846A CN103808846A (en) | 2014-05-21 |
| CN103808846B true CN103808846B (en) | 2015-04-29 |
Family
ID=50705922
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410057537.1A Expired - Fee Related CN103808846B (en) | 2014-02-20 | 2014-02-20 | Series quadrupole-rod gas-chromatographic mass spectrometry detection method for 35 toxic medicaments in urine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103808846B (en) |
Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572241B (en) * | 2014-11-05 | 2018-01-19 | 复旦大学 | The method for determining amphetamine in blood and urine, ketamine, pethidine and methadone |
| CN105987965A (en) * | 2015-02-09 | 2016-10-05 | 复旦大学 | Method for determining various types of abuse drugs in human whole blood |
| CN105651926A (en) * | 2016-04-15 | 2016-06-08 | 广西壮族自治区梧州食品药品检验所 | Method for detecting tetramine in urine of poisoning patients by gas chromatography-mass spectrometry |
| CN106248819A (en) * | 2016-07-13 | 2016-12-21 | 浙江景嘉医疗科技有限公司 | The detection method of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel |
| CN106370745B (en) * | 2016-08-25 | 2019-08-02 | 广东电网有限责任公司电力科学研究院 | The detection method of aminopyrine in a kind of transformer oil |
| CN106139641B (en) * | 2016-08-25 | 2019-10-08 | 广东电网有限责任公司电力科学研究院 | The processing method of aminopyrine in a kind of transformer oil |
| CN107478757B (en) * | 2017-05-08 | 2019-12-06 | 公安部物证鉴定中心 | Method for purifying and preparing ketamine standard substance for forensic science drug detection |
| CN107478747B (en) * | 2017-05-10 | 2019-11-08 | 山东省公安厅 | The liquid chromatography-mass spectrography screening method of unknown poisonous substance in blood |
| CN107192788B (en) * | 2017-05-10 | 2019-04-19 | 山东省公安厅 | The gas chromatography-mass spectrum screening method of unknown poisonous substance in blood |
| CN108760906A (en) * | 2018-04-12 | 2018-11-06 | 公安部物证鉴定中心 | A kind of detection method of new psychoactive drug substance pFPP |
| CN109828051B (en) * | 2019-03-11 | 2022-09-09 | 芜湖市疾病预防控制中心 | Method for detecting toxic compound |
| CN110346470A (en) * | 2019-07-04 | 2019-10-18 | 公安部物证鉴定中心 | A kind of detection method of 3,4- methylene-dioxy methcathinone |
| CN111751470B (en) * | 2020-07-07 | 2023-05-05 | 多多药业有限公司 | Detection control method for new impurities in tramadol hydrochloride preparation |
| CN113588830A (en) * | 2021-08-03 | 2021-11-02 | 四川大学 | Screening identification and confirmation standard, screening method and kit for common toxicants |
| CN113406244A (en) * | 2021-08-03 | 2021-09-17 | 四川大学 | Common poison screening database and rapid screening method based on liquid chromatogram-rod orbit trap mass spectrum |
| CN117434186A (en) * | 2023-12-22 | 2024-01-23 | 中国市政工程华北设计研究总院有限公司 | Synchronous identification and quantification method for complex odor gas composition of sewage-carrying rainwater |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128905A (en) * | 2010-12-10 | 2011-07-20 | 中国广州分析测试中心 | Method for quickly detecting drug |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050054942A1 (en) * | 2002-01-22 | 2005-03-10 | Melker Richard J. | System and method for therapeutic drug monitoring |
| US7585680B2 (en) * | 2003-08-28 | 2009-09-08 | Marshfield Medical Research And Education Foundation | Method and device for monitoring medication usage |
-
2014
- 2014-02-20 CN CN201410057537.1A patent/CN103808846B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128905A (en) * | 2010-12-10 | 2011-07-20 | 中国广州分析测试中心 | Method for quickly detecting drug |
Non-Patent Citations (1)
| Title |
|---|
| 分散固相萃取-气相色谱-三重四极杆质谱分析蔬菜中112 种农药残留;施家威等;《色谱》;20120630;第30卷(第6期);602-612 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103808846A (en) | 2014-05-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103808846B (en) | Series quadrupole-rod gas-chromatographic mass spectrometry detection method for 35 toxic medicaments in urine | |
| CN102200530B (en) | Method for detecting 33 kinds of narcotic drugs in urine by liquid chromatography-tandem mass spectrometry | |
| Yan et al. | Rapid and global detection and characterization of aconitum alkaloids in Yin Chen Si Ni Tang, a traditional Chinese medical formula, by ultra performance liquid chromatography–high resolution mass spectrometry and automated data analysis | |
| Wylie et al. | Drugs in oral fluid: Part I. Validation of an analytical procedure for licit and illicit drugs in oral fluid | |
| Ye et al. | Chemical fingerprinting of Liuwei Dihuang Pill and simultaneous determination of its major bioactive constituents by HPLC coupled with multiple detections of DAD, ELSD and ESI-MS | |
| CN103808840B (en) | A kind of method for building up of ginseng and astragalus injection for strengthening body finger-print | |
| Duan et al. | LC–MS/MS determination and pharmacokinetic study of five flavone components after solvent extraction/acid hydrolysis in rat plasma after oral administration of Verbena officinalis L. extract | |
| CN102226794B (en) | Liquid chromatography-tandom mass spectrometry detection method of thirty-one drugs in human blood | |
| Duan et al. | Study on the destructive effect to inherent quality of Fritillaria thunbergii Miq.(Zhebeimu) by sulfur-fumigated process using chromatographic fingerprinting analysis | |
| CN102879486A (en) | Method for screening traditional Chinese medicine effect related ingredients and model building method | |
| CN104991019B (en) | Gelsemine and the liquid chromatography-tandem mass of koumine in biological material | |
| Yang et al. | Internal standard mass spectrum fingerprint: a novel strategy for rapid assessing the quality of Shuang-Huang-Lian oral liquid using wooden-tip electrospray ionization mass spectrometry | |
| CN103235050B (en) | Quality control method of panax notoginseng saponins injection | |
| CN104833743A (en) | Method for analyzing cathinone, methcathinone and 4-methylmethcathinone in biological sample by liquid chromatography-mass spectrometry | |
| Jin et al. | Simultaneous quantification of 19 diterpenoids in Isodon amethystoides by high-performance liquid chromatography–electrospray ionization tandem mass spectrometry | |
| CN109444290B (en) | Construction method and detection method of UPLC (ultra performance liquid chromatography) characteristic map of plantain herb | |
| CN105181855A (en) | Method for simultaneously determining contents of 10 chemical components in fourstamen stephania root and astragalus membranaceus decoction preparation by UHPLC-MS/MS (Ultra High Performance Liquid Chromatography-Mass Spectrograph) | |
| CN109374764B (en) | HPLC fingerprint spectrum and main component content determination method for eight-ingredient Longzui granules | |
| Liu et al. | More accurate matrix-matched quantification using standard superposition method for herbal medicines | |
| Li et al. | Ultraviolet spectroscopy combined with ultra-fast liquid chromatography and multivariate statistical analysis for quality assessment of wild Wolfiporia extensa from different geographical origins | |
| Wang et al. | TCM-ADMEpred: a novel strategy for poly-pharmacokinetics prediction of traditional Chinese medicine based on single constituent pharmacokinetics, structural similarity, and mathematical modeling | |
| CN105954422A (en) | Method for quick detection of content of citrinin in traditional Chinese medicinal materials | |
| Liu et al. | Metabolomic study of a rat fever model induced with 2, 4-dinitrophenol and the therapeutic effects of a crude drug derived from Coptis chinensis | |
| CN104407082B (en) | The detection method of alkylphenol compounds in a kind of ginkgo leaf raw material and preparation | |
| CN105866295A (en) | Method for quickly detecting aflatoxin B1 content in traditional Chinese medicinal materials |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150429 |