CN103808846A - 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
- CN103808846A CN103808846A CN201410057537.1A CN201410057537A CN103808846A CN 103808846 A CN103808846 A CN 103808846A CN 201410057537 A CN201410057537 A CN 201410057537A CN 103808846 A CN103808846 A CN 103808846A
- Authority
- CN
- China
- Prior art keywords
- urine
- kinds
- poison
- medicine
- thing
- 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.)
- Granted
Links
Images
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.Once people is infected with among the deformity " pleasant sensation " that drugs will often wallow in drugs and had fantasies of out, 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 social serious harm, abroad some countries are except national drug addict is taked corresponding control measures, immigrant are also required to the proof that provides 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.Relate to health status aspect when responding actively more external countries to my personnel's entry visa and propose to add the requirement of doing illicit drugs inspection, inspection and quarantine department will be according to the requirement of State Bureau, actively carries out on the one hand material and technical preparation to tackle the external requirement to my entry personnel's health examination; From protection China's people's health and the object of maintaining social stability, also should take reciprocity principle on the other hand, the personnel of coming to China be increased to the requirement of illicit drugs inspection.
The measuring technology of the interior drug ingredient check of the most widely used human body is both at home and abroad immunoassay technology and vapor-phase chromatography (GC) at present, abroad also has (GC/MS) that use gas chromatography mass spectrometry to detect.Retrieval Chinese Industrial Standards (CIS) net is found, in the industry standard about a few class drugs such as morphine, ***e, heroin of the standard Zhong Jinyou Ministry of Public Security, is mainly the directly detection to drugs, and is not the detection method for clinical sample aspect.The method that inspection and quarantine department carries out illicit drugs inspection to inward and outward personnel's urine at present mainly adopts colloidal gold technique fast detecting morphine and meth, and false positive rate is high, and can only detect 2 kinds of drugs.Therefore foundation one is quick, sensitive, multiple poison (medicine) thing Simultaneous Detection becomes particularly important accurately.
China Patent No. is 201110075177.4 to disclose the Liquid Chromatography-Tandem Mass Spectrometry detection method of 33 kinds of drugs in a kind of urine, and its step is as follows:
Sample preparation: take 2g urine sample to be checked, be accurate to 0.01g, be placed in 50mL tool plug centrifuge tube, add 10ml ethyl acetate, on liquid blending device, fully mix 1min, with the centrifugal 10min of 4000r/min, draw upper strata ethyl acetate in test tube, then add 1mL0.1mol/L NaOH solution, after fully mixing on liquid blending device, add 10ml ethyl acetate to repeat to extract once, merge extracted twice liquid; Extract dries up instrument with nitrogen and dries up at 40 ℃, and adding 1mL volume ratio is the methyl alcohol of 30: 70: eddies of water supination, cross 0.25 μ m filter membrane, and obtain testing sample;
Measure: testing sample liquid chromatography series connection quadrupole mass spectrometer is measured; 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 spectrum condition of positive ion mode is: ionization source: ESI source; Scan pattern: positive ion is selected scan pattern SRM; Electron spray voltage: 2.5kV; Dry gas temperature: 400 ℃; Gas curtain atmospheric pressure: 45arb; Assisted gas pressure: 7arb; Capillary temperature: 350 ℃; Collision gas: high-purity argon gas;
The mass spectrum condition of negative ion mode is: ionization source: ESI source; Scan pattern: negative ion is selected scan pattern SRM; Electron spray voltage: 2.5kV; Dry gas temperature: 400 ℃; Gas curtain atmospheric pressure: 50arb; Assisted gas pressure: 7arb; Capillary temperature: 320 ℃; Collision gas: high-purity argon gas;
Chromatographic condition is: chromatographic column: Phenomenex Gemini C18100 × 2.0,3 μ m; Column temperature: room temperature; Flow velocity: 250 μ L/min; Sampling volume: 25 μ L.
China Patent No. is that its urine of documents of 201110075177.4 adopts classical liquid-liquid extraction method to carry out sample pre-treatments, adopts ethyl acetate as extraction solvent, and extractant shares amount 20ml, and can only detect 33 kinds of drugs.This method and adopt Liquid Chromatography-Tandem Mass Spectrometry to detect, the weak point of its method is, due to detection material some with positive ion scanning, some scans for negative ion.So will be divided into and detect the disposable detection that can not reach real for twice while detection.
Summary of the invention
The object of this invention is to provide a kind of 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 that adopts.
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) thing in described urine, it is characterized in that: the standard items of first using 35 kinds of poison (medicine) are set up multiple reaction detection method in series connection level Four bar gas chromatograph-mass spectrometer, determine 2～3 pairs of feature parent ions and daughter ion to and the retention time of every 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: m) quartz capillary column of DB-1MS(30m × 0.25mm × 0.25 μ, injector temperature: 250 ℃, do not shunt; Temperature programme: 50 ℃ of initial temperatures, are warmed up to 290 ℃ with 12 ℃/min and keep 10min, flow velocity: 1ml/min, sample size: 0.5 μ L; MS condition: EI ion gun, electronics energy 70eV, 230 ℃ of ion source temperatures, interface temperature: 280 ℃, level Four bar temperature: 150 ℃, electron-multiplier voltage: 1381eV.
Described series connection level Four bar gas chromatography combined with mass spectrometry detection method, specifically comprises:
1) standard solution 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 preparation
Get urine 1ml to be checked, add ether 6ml to divide three extractions, each 2ml; At 40 ℃ 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 ℃, nitrogen dries up, and residue dissolves with 100 μ L methyl alcohol, 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 ℃, do not shunt; Temperature programme: 50 ℃ of initial temperatures, are warmed up to 290 ℃ with 12 ℃/min and keep 10min, flow velocity: 1ml/min, sample size: 0.5 μ L;
2.3) MS condition
MS condition: EI ion gun, electronics energy 70eV, 230 ℃ of ion source temperatures, interface temperature: 280 ℃, level Four bar temperature: 150 ℃, 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: take the relative retention time of each drugs and the right abundance ratio of the monitoring parent ion/daughter ion of each drugs as qualitative foundation, the retention time of each drugs sees the above table 1; Quantitative by external standard method, with its residual quantity in sample of the calculated by peak area of each drugs;
4) matrix effect
By after above-mentioned " 2.1 " step process, add the standard solution of 35 kinds of poison (medicine) thing in blank urine sample, carry out GC-MS/MS analysis, obtain standard items peak area A; Equally the standard solution of 35 kinds of poison (medicine) thing is analyzed, obtained corresponding peak area B; Matrix effect=A/B × 100%, result show except nitrazepam and Clonazepam be that other 33 kinds of cytotoxic drugs of substrate inhibition are all matrix enhancements, 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) typical curve and detectability
Get blank urine sample, add 35 kinds of poison (medicine) thing standard solution to mix, 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 solution of 2.0,3.0,4.0,4.5,5.0,6.5 μ g/mL; By carrying out GC-MS/MS analysis after 2.1 step method pre-treatments; To cytotoxic drug mass concentration (C, μ g/mL) drawing standard curve, and try to achieve linear regression equation and related coefficient with cytotoxic drug peak area (A); The urine intoxicant substrate concentration (with 3 times snr 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;
Typical curve equation, related coefficient and the detection limit of table 335 kind of poison (medicine) thing
The main technical content of detection method of the present invention and ultimate principle:
1) extraction solvent contrast cyclohexane, ethyl acetate, four kinds of methenyl choloride and ether, find that extracted by ether rate is high, and ether boiling point is low, more concentrated, therefore selects ether as extraction solvent.
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 further verify with extracting ion pair, can detect the 7 large classes such as acephatemet, Tetramine, diazepam, nikethamidum, lidocaine, phenobarbital, atropine common poison (medicine) thing in totally 35 simultaneously, the recovery reaches 80.82% to 118.56%, and detection limit reaches 0.0014～1.8 μ g/ml.Chromatographic retention for this law, mass spectral characteristic parent ion/daughter ion be to simultaneously qualitative, with its residual quantity in sample of the calculated by peak area of each cytotoxic drug.The method has been eliminated the interference of complicated substrate in urine to greatest extent, can quick and precisely carry out qualitative and quantitative analysis to 35 kinds of poison (medicine) thing, widely applicable, simple to operate, detect with strong points.Can be used for inspection and quarantining for import/export, disease prevention and control center, the confirmation of public security department's fast detecting and positive findings.
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 concentrated, therefore select ether as extraction solvent; 2) documents that routine techniques is 201110075177.4 as the patent No. adopts high performance liquid chromatography-tandem mass technology, it can only detect 33 kinds of poison (medicine) thing, the present invention adopts simultaneously common poison (medicine) thing in 35 in Rapid Screening human urine of series connection level Four bar combined gas chromatography mass spectrometry (GC-MS/MS), assay is accurate, sensitive, quick, poison (medicine) thing 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 model GC-MS/MS multiple-reaction monitoring spectrograms.The retention time of 35 kinds of cytotoxic drugs and go out peak position as can be seen from FIG..
Embodiment
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) thing 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 methenyl choloride, ether, phosphoric acid, sodium dihydrogen phosphate, NaOH, sodium carbonate, sodium bicarbonate, hydrochloric acid etc. are analysis; Methyl alcohol is chromatographically pure.
2.3 standard solution preparations: 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 approachs
3.1 sample preparation
Get urine 1ml to be checked, add ether 6ml to divide three extractions, each 2ml.At 40 ℃ 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 ℃, nitrogen dries up, and residue dissolves with 100 μ L methyl alcohol, gets 1 μ L and enters GC/MS/MS analysis.
3.2GC condition
GC condition: DB-1 (m) quartz capillary column of 30m × 0.25mm × 0.25 μ, injector temperature: 250 ℃, do not shunt.Temperature programme: 50 ℃ of initial temperatures, are warmed up to 290 ℃ with 12 ℃/min and keep 10min, flow velocity: 1ml/min, sample size: 0.5 μ L.
3.3MS condition
MS condition: EI ion gun, electronics energy 70eV, 230 ℃ of ion source temperatures, interface temperature: 280 ℃, level Four bar temperature: 150 ℃, electron-multiplier voltage: 1381eV.
4 method optimization and method validations
4.1 sample pre-treatments optimizations
4.1.1 the optimization of extraction solvent
Extraction solvent contrast cyclohexane, ethyl acetate, four kinds of methenyl choloride and ether, find that extracted by ether rate is high, and ether boiling point is low, more concentrated, therefore selects ether as extraction solvent.
4.1.2 extract the optimization of pH value
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 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 composition well be separated.
The optimization of 4.3MS condition
4.3.1 determine the feature parent ion daughter ion pair of compound to be checked
First use the standard items of 35 kinds of poison (medicine) thing on GC/MS/MS, to set up multiple reaction monitoring (MRM) method, determine that the feature three of compound to be checked is to parent ion and daughter ion pair.
4.3.2 the exploitation of sectional detecting method
Detect the hybrid standard product of 35 kinds of poison (medicine) thing by this MRM method, suitably optimizing chromatographic condition makes 35 kinds of poison (medicine) thing substantially can reach baseline separation respectively, because can reaching baseline separation substantially, each thing to be checked therefore adopts each material to be divided into one to analyze the period, be chosen in the feature MRM method that this section go out the cytotoxic drug composition at peak and detect.The final retention time and feature parent ion and daughter ion used is to qualitative.When two materials are in the time that same retention time goes out peak or appearance time and differs very little, can adopt and extract parent ion daughter ion chromatogram is carried out to the qualitative and Quantitative Separation analysis of compound.
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.
5.235 kinds of cytotoxic drugs spectrogram in the time of optimum MRM method and sectional detecting method is shown in Fig. 1, and Figure 135 plants cytotoxic drug standard model GC-MS/MS multiple-reaction monitoring spectrogram, as can be seen from FIG. the retention time of 35 kinds of cytotoxic drugs and go out peak position.
5.3 qualitative-and-quantitative methods: take the relative retention time of each drugs and the right abundance ratio of the monitoring parent ion/daughter ion of each drugs as qualitative foundation, the retention time of each drugs sees the above table; Quantitative by external standard method, with its residual quantity in sample of the calculated by peak area of each drugs.
5.4 specificity
Due to the advance of GC-MS/MS method, adopt the feature parent ion daughter ion of target analytes to carrying out qualitative and quantitative analysis, can farthest get rid of matrix interference and improve the specificity that object is detected.
5.5 matrix effect
After blank urine sample is processed by " 3.1 ", add the standard solution of 35 kinds of poison (medicine) thing, carry out GC-MS/MS analysis, obtain standard items peak area A; Equally the standard solution of 35 kinds of poison (medicine) thing is analyzed, obtained corresponding peak area B; Matrix effect=A/B × 100%.Result show except nitrazepam and Clonazepam be that other 33 kinds of cytotoxic drugs of substrate inhibition are all matrix enhancements, 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 typical curves and detectability
Get blank urine sample, add 35 kinds of poison (medicine) thing standard solution to mix, 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 solution of 2.0,3.0,4.0,4.5,5.0,6.5 μ g/mL.By carrying out GC-MS/MS analysis after 3.1 method pre-treatments.To cytotoxic drug mass concentration (C, μ g/mL) drawing standard curve, and try to achieve linear regression equation and related coefficient with cytotoxic drug peak area (A).The urine intoxicant substrate concentration (with 3 times snr 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.
Typical curve equation, related coefficient and the detection limit of table 335 kind of poison (medicine) thing
5.7 accuracy and precision
By step " preparations of a 4.5 typical curves " lower operation, prepare quality control (QC) sample of basic, normal, high three concentration of 35 kinds of poison (medicine) thing, each concentration is carried out 5 times and is analyzed, METHOD FOR CONTINUOUS DETERMINATION five days, according to the typical curve on the same day, calculate the concentration that records of QC sample, calculate the veracity and precision of this law according to QC sample result, the results are shown in Table 4.
In a few days and in the daytime the recovery and the precision of table 435 kind of poison (medicine) thing
6 full patterns are measured
Get actual sample, wherein knownly contain DDVP, Tetramine, amytal, ketamine, doxepin and 6 kinds of poison of Clozapine (medicine) thing, and the concentration of known six kinds of poisonous substances is 5 μ g/mL.This actual sample is processed according to step " 3.1 " sample-pretreating method, carries out GC-MS/MS analysis, and measurement result is in table 5.As known from Table 5, adopt in detection method actual sample of the present invention 6 kinds of poison (medicine) thing all accurately qualitative, the relative deviation of the content obtaining by quantitative curve 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 is measured the result table of poison (medicine) thing content
| Cytotoxic drug title | Retention time (t/min) | Detected value (μ g/mL) | Relative deviation (%) |
| DDVP | 8.246 | 5.1931 | 3.86 |
| Tetramine | 12.459 | 4.8989 | 2.02 |
| Amytal | 13.128 | 5.1822 | 3.64 |
| Ketamine | 14.448 | 4.9018 | 1.96 |
| Doxepin | 17.524 | 4.8093 | 3.81 |
| Clozapine | 21.993 | 5.2158 | 4.31 |
7 discuss
The present invention utilizes GC-MS/MS to have the advantages such as very high specificity, sensitivity, accuracy, set up the disposable new method that simultaneously detects 35 kinds of common cytotoxic drugs, carry out system thinking and optimization to extracting the sample pre-treatments such as solvent, system pH, chromatographic column and testing conditions, determined that the optimum parent ion/daughter ion of 35 kinds of poison is to (specifically in table 1).Under selected condition, further provide detectability (specifically in table 3) 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), every kind of material etc., 35 kinds of cytotoxic drug institute method for building up in a few days and in the daytime the 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, can disposablely carry out qualitative and quantitative analysis to 35 kinds of cytotoxic drugs simultaneously, simultaneously qualitative with chromatographic retention and mass spectrum, eliminating other composition in urine disturbs, result accurately and reliably, selectivity and reproducible, actual sample qualitative and quantitative detection all obtains satisfactory result, shows that the GC-MS/MS method of setting up can meet the requirement of illicit drugs inspection in blood of human body and urine accurately and reliably.
Claims (2)
1. the series connection level Four bar gas chromatography combined with mass spectrometry detection method of 35 kinds of poison (medicine) thing in a urine, it is characterized in that: the standard items of first using 35 kinds of poison (medicine) are set up multiple reaction detection method in series connection level Four bar gas chromatograph-mass spectrometer, determine 2～3 pairs of feature parent ions and daughter ion to and the retention time of every 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: m) quartz capillary column of DB-1MS(30m × 0.25mm × 0.25 μ, injector temperature: 250 ℃, do not shunt; Temperature programme: 50 ℃ of initial temperatures, are warmed up to 290 ℃ with 12 ℃/min and keep 10min, flow velocity: 1ml/min, sample size: 0.5 μ L; MS condition: EI ion gun, electronics energy 70eV, 230 ℃ of ion source temperatures, interface temperature: 280 ℃, level Four bar temperature: 150 ℃, electron-multiplier voltage: 1381eV.
2. series connection level Four bar gas chromatography combined with mass spectrometry detection method according to claim 1, is characterized in that:
1) standard solution 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 preparation
Get urine 1ml to be checked, add ether 6ml to divide three extractions, each 2ml; At 40 ℃ 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 ℃, nitrogen dries up, and residue dissolves with 100 μ L methyl alcohol, 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 ℃, do not shunt; Temperature programme: 50 ℃ of initial temperatures, are warmed up to 290 ℃ with 12 ℃/min and keep 10min, flow velocity: 1ml/min, sample size: 0.5 μ L;
2.3) MS condition
MS condition: EI ion gun, electronics energy 70eV, 230 ℃ of ion source temperatures, interface temperature: 280 ℃, level Four bar temperature: 150 ℃, 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: take the relative retention time of each drugs and the right abundance ratio of the monitoring parent ion/daughter ion of each drugs as qualitative foundation, the retention time of each drugs sees the above table 1; Quantitative by external standard method, with its residual quantity in sample of the calculated by peak area of each drugs;
4) matrix effect
By after above-mentioned " 2.1 " step process, add the standard solution of 35 kinds of poison (medicine) thing in blank urine sample, carry out GC-MS/MS analysis, obtain standard items peak area A; Equally the standard solution of 35 kinds of poison (medicine) thing is analyzed, obtained corresponding peak area B; Matrix effect=A/B × 100%, result show except nitrazepam and Clonazepam be that other 33 kinds of cytotoxic drugs of substrate inhibition are all matrix enhancements, 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) typical curve and detectability
Get blank urine sample, add 35 kinds of poison (medicine) thing standard solution to mix, 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 solution of 2.0,3.0,4.0,4.5,5.0,6.5 μ g/mL; By carrying out GC-MS/MS analysis after 2.1 step method pre-treatments; To cytotoxic drug mass concentration (C, μ g/mL) drawing standard curve, and try to achieve linear regression equation and related coefficient with cytotoxic drug peak area (A); The urine intoxicant substrate concentration (with 3 times snr 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;
Typical curve equation, related coefficient and the detection limit of table 335 kind of poison (medicine) thing
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 true CN103808846A (en) | 2014-05-21 |
| CN103808846B 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) |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572241A (en) * | 2014-11-05 | 2016-05-11 | 复旦大学 | Method for assaying amphetamines, ketamine, pethidine and methadone in blood and urine |
| 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 |
| CN105987965A (en) * | 2015-02-09 | 2016-10-05 | 复旦大学 | Method for determining various types of abuse drugs in human whole blood |
| CN106139641A (en) * | 2016-08-25 | 2016-11-23 | 广东电网有限责任公司电力科学研究院 | The processing method of aminophenazone in a kind of transformer oil |
| 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 |
| CN106370745A (en) * | 2016-08-25 | 2017-02-01 | 广东电网有限责任公司电力科学研究院 | Detection method of aminopyrine in transformer oil |
| CN107192788A (en) * | 2017-05-10 | 2017-09-22 | 山东省公安厅 | The gaschromatographic mass spectrometry screening method of unknown poisonous substance in blood |
| CN107478757A (en) * | 2017-05-08 | 2017-12-15 | 公安部物证鉴定中心 | Method for preparing purified for the ketamine standard substance of forensic science illicit drugs inspection |
| CN107478747A (en) * | 2017-05-10 | 2017-12-15 | 山东省公安厅 | The liquid chromatography mass 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 |
| CN109828051A (en) * | 2019-03-11 | 2019-05-31 | 芜湖市疾病预防控制中心 | A kind of detection method of toxic compounds |
| 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 |
| CN111751470A (en) * | 2020-07-07 | 2020-10-09 | 多多药业有限公司 | Detection control method for new impurities in tramadol hydrochloride preparation and impurities obtained by detection control method |
| 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 |
| CN113588830A (en) * | 2021-08-03 | 2021-11-02 | 四川大学 | Screening identification and confirmation standard, screening method and kit for common toxicants |
| 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 (3)
| 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 |
| US20110086428A1 (en) * | 2003-08-28 | 2011-04-14 | Larson Michael E M | Method and device for monitoring medication usage |
| CN102128905A (en) * | 2010-12-10 | 2011-07-20 | 中国广州分析测试中心 | Method for quickly detecting drug |
-
2014
- 2014-02-20 CN CN201410057537.1A patent/CN103808846B/en not_active Expired - Fee Related
Patent Citations (3)
| 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 |
| US20110086428A1 (en) * | 2003-08-28 | 2011-04-14 | Larson Michael E M | Method and device for monitoring medication usage |
| 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 |
|---|
| 施家威等: "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", 《色谱》, vol. 30, no. 6, 30 June 2012 (2012-06-30), pages 602 - 612 * |
Cited By (22)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572241A (en) * | 2014-11-05 | 2016-05-11 | 复旦大学 | Method for assaying amphetamines, ketamine, pethidine and methadone in blood and urine |
| 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 |
| CN106139641A (en) * | 2016-08-25 | 2016-11-23 | 广东电网有限责任公司电力科学研究院 | The processing method of aminophenazone in a kind of transformer oil |
| CN106370745A (en) * | 2016-08-25 | 2017-02-01 | 广东电网有限责任公司电力科学研究院 | Detection method of aminopyrine in 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 |
| CN107478757A (en) * | 2017-05-08 | 2017-12-15 | 公安部物证鉴定中心 | Method for preparing purified for the ketamine standard substance of forensic science illicit drugs inspection |
| 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 |
| CN107478747A (en) * | 2017-05-10 | 2017-12-15 | 山东省公安厅 | The liquid chromatography mass screening method of unknown poisonous substance in blood |
| CN107192788A (en) * | 2017-05-10 | 2017-09-22 | 山东省公安厅 | The gaschromatographic mass spectrometry 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 |
| CN109828051A (en) * | 2019-03-11 | 2019-05-31 | 芜湖市疾病预防控制中心 | A kind of detection method of toxic compounds |
| 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 |
| CN111751470A (en) * | 2020-07-07 | 2020-10-09 | 多多药业有限公司 | Detection control method for new impurities in tramadol hydrochloride preparation and impurities obtained by detection control method |
| 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 |
| CN113588830A (en) * | 2021-08-03 | 2021-11-02 | 四川大学 | Screening identification and confirmation standard, screening method and kit for common toxicants |
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103808846B (en) | 2015-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103808846B (en) | Series quadrupole-rod gas-chromatographic mass spectrometry detection method for 35 toxic medicaments in urine | |
| Wylie et al. | Drugs in oral fluid: Part I. Validation of an analytical procedure for licit and illicit drugs in oral fluid | |
| CN103808840B (en) | A kind of method for building up of ginseng and astragalus injection for strengthening body finger-print | |
| CN102200530B (en) | Method for detecting 33 kinds of narcotic drugs in urine by liquid chromatography-tandem mass spectrometry | |
| 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 | |
| Duan et al. | Study on the destructive effect to inherent quality of Fritillaria thunbergii Miq.(Zhebeimu) by sulfur-fumigated process using chromatographic fingerprinting analysis | |
| CN102226794B (en) | Liquid chromatography-tandom mass spectrometry detection method of thirty-one drugs in human blood | |
| Stephanson et al. | Method validation and application of a liquid chromatography–tandem mass spectrometry method for drugs of abuse testing in exhaled breath | |
| CN102879486A (en) | Method for screening traditional Chinese medicine effect related ingredients and model building method | |
| 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 | |
| CN104991019B (en) | Gelsemine and the liquid chromatography-tandem mass of koumine in biological material | |
| CN107607649B (en) | Method for detecting periploca forrestii schltr | |
| 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 | |
| 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) | |
| Lu et al. | Detection of ***e and its metabolites in urine using solid phase extraction-ion mobility spectrometry with alternating least squares | |
| 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 | |
| CN102539597B (en) | Method for quickly identifying notopterygium incisum seed and notopterygium franchetii seed | |
| CN107529337A (en) | The HPLC analyses of impurity in two to the water wei ling alcohol | |
| 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 | |
| Song et al. | Simultaneous quantification of 16 bioactive constituents in common Cnidium fruit by liquid chromatography–electrospray ionization-mass spectrometry | |
| CN102507794A (en) | Method for rapidly detecting poisoned substances by adopting urine |
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 |