CN108181385A - Method that is a kind of while detecting the enantiomter of crystal methamphetamine and amphetamine in liquid biological sample - Google Patents
Method that is a kind of while detecting the enantiomter of crystal methamphetamine and amphetamine in liquid biological sample Download PDFInfo
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- CN108181385A CN108181385A CN201711354278.9A CN201711354278A CN108181385A CN 108181385 A CN108181385 A CN 108181385A CN 201711354278 A CN201711354278 A CN 201711354278A CN 108181385 A CN108181385 A CN 108181385A
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- G01N30/02—Column chromatography
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The present invention provides a kind of methods for detecting R () crystal methamphetamine in liquid biological sample, S (+) crystal methamphetamine, R () amphetamine and S (+) amphetamine simultaneously, include the following steps：(1) sample pre-treatments：At room temperature, liquid biological sample sample is taken, is placed in an EP pipes, and add in inner mark solution；Implement first time vortex oscillation, be then centrifuged for, supernatant taken to be placed in another EP pipes, and add in methanol；Implement second of vortex oscillation, finally, sample introduction to LC MS/MS systems；(2) LC MS/MS are analyzed.Method of the present invention has excellent selectivity, linear dependence, preci-sion and accuracy, LOD and LOQ is sufficiently low simultaneously, therefore, it is very suitable for the chiral separation and measure of crystal methamphetamine and the respective enantiomter of amphetamine in blood, urine, success is speculates that drug source and the illegal abuse of correct differentiation and clinical application provide technical support, so as to have considerable market potential.
Description
Technical field
The invention belongs to field of chemical detection, and in particular to a kind of to detect R (-)-methylbenzene in liquid biological sample simultaneously
Propylamine, S (+)-crystal methamphetamine, R (-)-amphetamine and S (+)-amphetamine method.
Background technology
Crystal methamphetamine (methamphetamine, MA) also known as methamphetamine or removes (de-) oxygen ephedrine (alkali),
Its hydrochloride is a kind of transparent crystal, is commonly called as ice；Amphetamine (amphetamine, AM) is also known as amphetamine, is ephedrine
Derivative；The two belongs to amphetamine central nervous excitation agent, for country《Psychotropic substances kind catalogue》Middle first kind essence
God's medicine product.Wherein, amphetamine can be used as the main activity in vivo metabolin of drugs exclusive use and crystal methamphetamine.
Crystal methamphetamine and amphetamine are chipal compounds, and the pharmacology of their enantiomter in vivo is lived
There are significant differences for property, metabolic process, metabolic rate and toxicity etc..For example, the pharmacological activity of S (+)-crystal methamphetamine compares R
Configuration is 5 times strong.S (+)-crystal methamphetamine has central nervous excitation effect, can be clinically used for treatment obesity；R (-)-first
The effect of base amphetamine central nervous excitation is weaker, and periphery sympatheticomimetic action is strong, clinically can be used as nasal vasoconstrictor and pa
The synthesis pro-drug of the gloomy sick medicine selegiline of gold.
The chiral separation of crystal methamphetamine and amphetamine, analysis have very big application value in forensic science field.
For example, the crystal methamphetamine drugs sold illegally in the market have plenty of S configurations, have plenty of R configurations, some is identical for S with R configurations
The raceme of ratio also has the mixture of different proportion.The crystal methamphetamine of S configurations is mainly by 1R, 2S- ephedrines or 1S, 2S-
Pseudoephedrine reduction generation, two kinds of precursor compounds are mainly extracted from Herba Ephedrae to be obtained.The crystal methamphetamine of R configurations is usual
By 1R, 2R Pseudoephedrines or the synthesis of 1S, 2R- ephedrine, and the precursor compound of the configuration is not contained in Herba Ephedrae.Raceme
Crystal methamphetamine is then mainly synthesized by benzyltrimethyl ketone through reduction amination or Leuckart reactions.In addition, high light purity
Crystal methamphetamine can be also further purified to obtain by racemic methyl amphetamine by chiral separation.Therefore, by methylbenzene
The analysis of propylamine enantiomter chiral ratio, people can further investigate the popularity of crystal methamphetamine abuse, place under arrest
The intrinsic property of drugs and the source of drugs, so as to provide technical support for the investigation of drug-related case, trial and identification.This
Outside, the chiral information of crystal methamphetamine is alternatively the synthesis offer thinking of certain compounds using it as precursor.
On the other hand, some clinical treatment drugs contain or are metabolized to crystal methamphetamine or pair of amphetamine various configuration
Body is reflected, and is drained by urine, is such as used for the prescription medicine selegiline (selegiline) for the treatment of of Parkinson disease；Selegiline mouth
It is absorbed after clothes rapidly, metabolism in vivo can generate R (-)-crystal methamphetamine, R (-)-amphetamine, S(+)DMS.Therefore, it is chiral
Crystal methamphetamine and the respective enantiomter of amphetamine are detached, exists in clinical, judicial expertise and toxicological aspect and focuses on
Want meaning.
In the prior art there are three types of the chiral separation methods of mainstream：Diastereomer is formed after chiral reagent derivatization again to divide
From；With chiral post separation；It is introduced into the Chiral Mobile Phase Additives to chromatographic system such as chiral cyclodextrin, to form temporary diastereomeric
Nanocrystal composition finally makes Chiral Separation.However, current chiral analysis focuses primarily upon clinical medicine, for methylbenzene third
The research report of amine and amphetamine is less, particularly, lacks the method for body fluid sample feature, the sample in existing method
Often dosage is larger, sensitivity is inadequate, specificity is not strong.
The chiral detection method of existing crystal methamphetamine includes：Gas chromatography-mass spectrometry (GC-MS), capillary
Electrophoresis (CE), high performance liquid chromatography (HPLC) and Liquid Chromatography-Mass Spectrometry LC-MS.Wherein, GC-MS has become wide
The technology of general application, but this method sample pre-treatments need derivatization step, it is cumbersome, time-consuming, laborious.There are many application sides by LC
Formula, but before chiral column is entered, sample pre-treatments are equally complicated and inconvenient；For example, it is desired to it is handed over strong cation
It changes column and crystal methamphetamine is extracted from urine, then derivatization is completed with chirality or achiral derivatization reagent.It is although near
LC-MS analyses are more and more important over year, but the application in crystal methamphetamine and the chiral detection field of amphetamine is still deficient.
The chiral detection method having built up is often there are shortcomings, for example, sensitivity is poor and acquisition time is longer.Therefore, it is
Detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-crystal methamphetamine, R (-)-amphetamine and S (+)-benzene simultaneously
Propylamine, it is urgent to provide chiral separation a kind of easy to operate and rapid, highly sensitive, high specificity and analysis methods.
Invention content
The present invention is directed to overcome defect in the prior art, and provide it is a kind of it is easy, fast and efficiently while detect
R (-)-crystal methamphetamine in liquid biological sample, S (+)-crystal methamphetamine, R (-)-amphetamine and S (+)-amphetamine method,
So as to speculate that drug source and the illegal abuse of correct differentiation and clinical application provide technical support.
Therefore, the present invention provides following technical schemes：
It is a kind of to detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-crystal methamphetamine, R (-)-amphetamine simultaneously
With the method for S (+)-amphetamine, include the following steps：
(1) sample pre-treatments；
(2) LC-MS/MS is analyzed；
Wherein, the sample pre-treatments described in step (1) include：
At room temperature, liquid biological sample sample is taken, is placed in an EP pipes, and add in inner mark solution；Implement first time whirlpool
Rotation oscillation, is then centrifuged for, supernatant is taken to be placed in another EP pipes, and add in methanol；Implement second of vortex oscillation, finally, into
Sample is to LC-MS/MS systems.
Preferably, in the above-mentioned methods, the liquid biological sample is blood or urine.
Preferably, in the above-mentioned methods, the inner mark solution is the methanol solution of 4- phenyibutylamines.
It is further preferred that in the above-mentioned methods, a concentration of 0.015 μ g/mL of the inner mark solution.
Preferably, in the above-mentioned methods, the concrete operations of the sample pre-treatments include：
At room temperature, 20 μ L liquid biological sample samples are taken, are placed in an EP pipes, and add in 980 μ L inner mark solutions；Implement
First time 15~20s of vortex oscillation, is then centrifuged for 1~2min, and 50 μ L supernatants is taken to be placed in another EP pipes, and adds in 450 μ L
Methanol；Implement second of 5~10s of vortex oscillation, finally, sample introduction to LC-MS/MS systems.
It is further preferred that in the above-mentioned methods, the volume of the EP pipes is 2mL.
It is further preferred that in the above-mentioned methods, the rotating speed of the centrifugation is 10000~13000r/min.
Preferably, in the above-mentioned methods, in the LC-MS/MS analyses described in step (2), chromatographic condition includes：
Chromatographic column is SupelcoAstec ChirobioticTMV2 chiral columns (2.1mm × 250mm, 5 μm)；Mobile phase：First
Alcohol/0.1% (v/v) glacial acetic acid/0.02% (v/v) ammonium hydroxide；Flow velocity：0.25mL/min；Sample size：20μL；Injector temperature：25
℃；Column temperature：25℃.
Preferably, in the above-mentioned methods, in the LC-MS/MS analyses described in step (2), Mass Spectrometry Conditions include：
Electron spray ionisation-positive ion mode (ESI+)；Collision gas (CAD)：7psi；Gas curtain gas (CUR)：30psi；Ion sprays
Radio pressure (IS)：5500V；Ion source gas 1 (GS1)：35psi；Ion source gas 2 (GS2)：35psi；Ion source temperature (TEM)：
500℃；Scan mode：More reaction detections (MRM).
Wherein, it detects ion and other relevant parameters see the table below 1：
The MS/MS conditions and retention time of 14 kinds of compounds of table
* the first pair of ion pair is for quantitative
Through method validation, R (-)-crystal methamphetamine, S (+)-crystal methamphetamine, R (-)-amphetamine and S (+)-amphetamine
The range of linearity is in 0.05-50.00 μ g/mL, the related coefficient (r of all standard curves2) it is all higher than 0.994；It is also, minimum
Detection limit (LOD) is 0.02 μ g/mL, and minimum quantitative limit (LOQ) is 0.05 μ g/mL.In addition, if actually detected result is more than mark
Directrix curve highest quantitative limit can will then measure after liquid biological sample sample methanol dilution.
Compared with prior art, technical solution provided by the present invention has the following advantages：
Liquid biological sample sample dilutes pre-treatment and the LC-MS/MS methods of chiral column combination are simple, quick and accurate；
Wherein, the dilution process of sample does not need to operating personnel and spends too many energy and time compared to extraction or derivatization method,
Therefore labour and time cost is greatly saved compared to GC-MS methods.The detection liquid biological inspection of the present invention while
R (-)-crystal methamphetamine in material, S (+)-crystal methamphetamine, R (-)-amphetamine and S (+)-amphetamine method in, simple sample
Product pre-treatment operate, significantly improve the working efficiency of large sample amount, and method accuracy and precision the result shows that, the inspection
Survey method is accurately and reliably.In addition, compared to other LC-MS methods up at least acquisition time of 35min, it is provided by the present invention
The complete baseline separation between crystal methamphetamine and each enantiomter of amphetamine only can be realized in method in 15min.Separately
Outside, it compared to the cutoff that 1mL urines to be measured is at least needed to can be only achieved standard requirement in existing literature method, invents described
Detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-crystal methamphetamine, R (-)-amphetamine and S (+)-phenylpropyl alcohol simultaneously
The method of amine has been obviously improved the sensitivity of detection method, therefore liquid biological because employing complete LC-MS/MS systems
The sampling amount of sample only needs 20 μ L.
In conclusion R (-)-crystal methamphetamine, S (+)-methyl in liquid biological sample are detected while of the present invention
The method of amphetamine, R (-)-amphetamine and S (+)-amphetamine has excellent selectivity, linear dependence, precision and accurate
Degree, while LOD and LOQ is sufficiently low, therefore, is very suitable for crystal methamphetamine and the respective mapping of amphetamine in blood, urine
The chiral separation and measure of isomers, successfully to speculate that drug source and the illegal abuse of correct differentiation and clinical application provide technology
It supports, so as to considerable market potential.
Description of the drawings
Fig. 1 is to detect the obtained chromatogram of blank diaper using the method for the invention；Wherein, horizontal axis coordinate is retains
Time, ordinate of orthogonal axes are abundance；
Fig. 2 is to be added to S (+)-crystal methamphetamine, R (-)-crystal methamphetamine, S using the method for the invention detection
The obtained chromatogram of the blank diaper of (+)-amphetamine and R (-)-amphetamine；S (+)-crystal methamphetamine, R (-) in the urine-
The respective concentration of crystal methamphetamine, S (+)-amphetamine and R (-)-amphetamine is 0.05 μ g/mL (LOQ)；Wherein, horizontal axis coordinate
For retention time, ordinate of orthogonal axes is abundance；
Fig. 3 is the obtained practical positive urine liquid chromatography of urine using the method for the invention detection person that sucks methamphetamine
Figure.
Specific embodiment
The present invention is further elaborated With reference to embodiment, but the present invention is not limited to following embodiment party
Formula.
It is a kind of to detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-crystal methamphetamine, R (-)-amphetamine simultaneously
With the method for S (+)-amphetamine, include the following steps：
(1) sample pre-treatments；
(2) LC-MS/MS is analyzed；
Wherein, the sample pre-treatments described in step (1) include：
At room temperature, liquid biological sample sample is taken, is placed in an EP pipes, and add in inner mark solution；Implement first time whirlpool
Rotation oscillation, is then centrifuged for, supernatant is taken to be placed in another EP pipes, and add in methanol；Implement second of vortex oscillation, finally, into
Sample is to LC-MS/MS systems.
In a preferred embodiment, the liquid biological sample is blood or urine.
In a preferred embodiment, the inner mark solution is the methanol solution of 4- phenyibutylamines.
In a further preferred embodiment, a concentration of 0.015 μ g/mL of the inner mark solution.
In a preferred embodiment, the concrete operations of the sample pre-treatments include：
At room temperature, 20 μ L liquid biological sample samples are taken, are placed in an EP pipes, and add in 980 μ L inner mark solutions；Implement
First time 15~20s of vortex oscillation, is then centrifuged for 1~2min, and 50 μ L supernatants is taken to be placed in another EP pipes, and adds in 450 μ L
Methanol；Implement second of 5~10s of vortex oscillation, finally, sample introduction to LC-MS/MS systems.
In a further preferred embodiment, the volume of the EP pipes is 2mL.
In a further preferred embodiment, the rotating speed of the centrifugation is 10000~13000r/min.
In a preferred embodiment, in the LC-MS/MS analyses described in step (2), chromatographic condition includes：
Chromatographic column is SupelcoAstec ChirobioticTMV2 chiral columns (2.1mm × 250mm, 5 μm)；Mobile phase：First
Alcohol/0.1% (v/v) glacial acetic acid/0.02% (v/v) ammonium hydroxide；Flow velocity：0.25mL/min；Sample size：20μL；Injector temperature：25
℃；Column temperature：25℃.
In a preferred embodiment, in the LC-MS/MS analyses described in step (2), Mass Spectrometry Conditions include：
Electron spray ionisation-positive ion mode (ESI+)；Collision gas (CAD)：7psi；Gas curtain gas (CUR)：30psi；Ion sprays
Radio pressure (IS)：5500V；Ion source gas 1 (GS1)：35psi；Ion source gas 2 (GS2)：35psi；Ion source temperature (TEM)：
500℃；Scan mode：More reaction detections (MRM).
In addition, a series of experiments is also embodied to be verified to the method for the invention in inventor, it is main to include verification
The selectivity, linear dependence and its quantitative limit of the method for the invention, preci-sion and accuracy etc.；It is as described below respectively：
Method choice
The 20 μ L of blank diaper of 10 parts of different peoples (health for not sucking methamphetamine is personal) are taken respectively, are carried out according to step (1)
Urine sample pre-treatment operates, and then implements LC-MS/MS analyses according to step (2), the results showed that, in compound (R to be analyzed
(-)-crystal methamphetamine, S (+)-crystal methamphetamine, R (-)-amphetamine and S (+)-amphetamine) it is equal at corresponding retention time window
No endogenous Interference Peaks occur (referring to Fig. 1), illustrate that the selectivity of this method is good.
Standard curve is limited with detection
The blank diaper for taking the health for not sucking methamphetamine personal is appropriate, is separately added into S (+)-crystal methamphetamine, R (-)-first
Base amphetamine, S (+)-amphetamine and R (-)-amphetamine series mixed standard solution are appropriate, are made and are equivalent to S (+)-methylbenzene third
Respectively concentration is respectively 0.05,0.15,0.40,2.00 for amine, R (-)-crystal methamphetamine, S (+)-amphetamine and R (-)-amphetamine,
The serial urine sample of 10.00 and 50.00 μ g/mL carries out urine sample pre-treatment operation, then according to step according to step (1)
Suddenly (2) implement LC-MS/MS analysis (S (+)-crystal methamphetamine, R (-)-crystal methamphetamine, S (+)-amphetamine and R (-)-phenylpropyl alcohol
Result when the respective concentration of amine is 0.05 μ g/mL is referring to Fig. 2, and the corresponding figure of same substance goes out peak position phase under other concentration
Closely, these attached drawings are omitted therefore).With test substance concentration in urine (μ g/mL) for abscissa x, test substance and internal standard compound
Peak area ratio is ordinate y, regressing calculation is carried out with weighting (w=1/x2) least square method, as a result with linear regression equation y
=ax+b is represented, see the table below 2：
The correction equation of 24 kinds of compounds of table
The above result shows that the range of linearity of test substance (4 kinds of compounds) is 0.05-50.00 μ g/mL, all marks
Related coefficient (the r of directrix curve2) 0.994 is both greater than, and its minimum quantitative limit (LOQ) is 0.05 μ g/mL.
Preci-sion and accuracy
The blank diaper for taking the health for not sucking methamphetamine personal is appropriate, prepares LOQ, basic, normal, high four concentration (S (+)-first
Base amphetamine, R (-)-crystal methamphetamine, S (+)-amphetamine and the respective concentration of R (-)-amphetamine are respectively 0.05,0.10,
0.50,40.00 μ g/mL) quality control (QC) sample, each concentration carries out 6 sample analyses, METHOD FOR CONTINUOUS DETERMINATION 4 days, according to the same day
Standard curve, calculate QC samples concentration.To measurement result carry out variance analysis, investigate method in a few days with day to day precision
(mean value of QC sample measured values is to the relatively inclined of sign value by RSD (relative standard deviation of QC sample measured values) and accuracy RE
Difference).Said determination result see the table below 3：
3 precision of table and accuracy
As it can be seen that R (-)-crystal methamphetamine, S (+)-methylbenzene third in liquid biological sample are detected while of the present invention
The absolute value of the accuracy RE of the method for amine, R (-)-amphetamine and S (+)-amphetamine is less than 7.1%, and in a few days and smart in the daytime
Density RSD is respectively smaller than 13.1% and 2.0%, shows that the accuracy of this method and precision are good, meets Determination of Biological Samples
Related request.
Procedure efficiency
The blank diaper and appropriate methanol for taking the health for not sucking methamphetamine personal, are separately added into S (+)-crystal methamphetamine, R
(-)-crystal methamphetamine, S (+)-amphetamine and R (-)-amphetamine series mixed standard solution, it is dense to be respectively prepared basic, normal, high three
S (+)-crystal methamphetamine of degree (0.10,0.50 and 40.00 μ g/mL), R (-)-crystal methamphetamine, S (+)-amphetamine and R (-)-
The QC samples (sample containing matrix) of amphetamine and reference sample (being free of matrix sample), sample pre-treatments are carried out according to step (1)
Then operation implements LC-MS/MS analyses according to step (2).Each concentration carries out 6 sample analyses, obtains corresponding peak area, and
Procedure efficiency is calculated by the peak area ratio of two groups of samples (A is free of matrix containing matrix/A).The result shows that S (+)-
Crystal methamphetamine, R (-)-crystal methamphetamine, S (+)-amphetamine and R (-)-amphetamine are corresponding under the conditions of various concentration
Ratio is between 85%~98%, RSD% ＜ 13%, therefore shows that this method process efficiency is stable and efficient, in detail number
According to the results are shown in Table 4：
4 procedure efficiency (n=6) of table
Residual effect
Untested compound (S (+)-crystal methamphetamine, R are completed with the 50.00 μ g/mL of maximum concentration in the range of linearity
(-)-crystal methamphetamine, S (+)-amphetamine and R (-)-amphetamine) test after, blank testing urine；It has been observed that blank is urinated
In liquid chromatography figure, residual peak is had no in the corresponding reservation period of untested compound.
Dilution effect
The blank diaper for taking the health for not sucking methamphetamine personal is appropriate, is separately added into S (+)-crystal methamphetamine, R (-)-first
Base amphetamine, S (+)-amphetamine and R (-)-amphetamine series mixed standard solution, are made and are equivalent to S (+)-crystal methamphetamine, R
The QC samples of (-)-crystal methamphetamine, S (+)-amphetamine and the respective a concentration of 200 μ g/mL of R (-)-amphetamine.According to step
(1) after carrying out sample pre-treatments operation, with 5 times of methanol dilution, implement LC-MS/MS analyses according still further to step (2), 20 μ L of sample introduction,
Record chromatogram, each 6 parts of concentration parallel determination.The concentration of each composition is obtained by standard curve, by the concentration measured with it is known
Addition concentrations versus, investigate with the accuracy of methanol dilution urine and precision；The result shows that urine sample methanol dilution
The accuracy RE absolute values of measurement result are respectively less than 8.0% after 5 times, and precision RSD is respectively less than 10%, therefore, if practical inspection
Result (or predicted concentration) is surveyed to be above standard curve highest quantitative limit, then it can be (such as practical positive by liquid biological sample sample
Urine) it is measured again with after methanol dilution.
With reference to specific embodiment, the present invention is further elaborated, but the present invention is not limited to following embodiments.
Embodiment 1
It is described while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-crystal methamphetamine, R (-)-amphetamine
Methylbenzene third in the blood in the internal sample quality assurance planning of certain law enforcement agency is applied to the method for S (+)-amphetamine
The assay project of amine and amphetamine.
Blood sample number to be checked is respectively BS-1, BS-2 and BS-3, and sample pre-treatments are carried out by the step (1), and
Three parts of operation repetitive respectively implements LC-MS/MS analyses then according to the step (2).Sample is calculated according to the standard curve on the same day
The concentration of S (+)-crystal methamphetamine that may contain in product, R (-)-crystal methamphetamine, S (+)-amphetamine and R (-)-amphetamine,
As a result 5 be see the table below：
The content of crystal methamphetamine and amphetamine in 5 blood sample of table
As it can be seen that do not detect R (-)-crystal methamphetamine in each blood sample, two kinds of enantiomers of measured amphetamine it is total
Concentration is consistent with the announcement result of certain law enforcement agency；Therefore testing result shows established method accurately and reliably.
Embodiment 2
The doubtful urine for abusing crystal methamphetamine personnel of the Yangtze River Delta Region of public security organ's inspection, collects and is moulding
Expect in bottle, be stored in refrigerator-freezer in -80 DEG C, and by the immune plate primary dcreening operation and laboratory GC-MS of the completion of other testing agencies
It is analyzed to identify as positive urine sample.Blank diaper separately is had, by (not sucking methamphetamine) healthy individuals will of no medication history
Hope person provides.
Be collected into 86 parts of positive urine samples are subjected to sample pre-treatments and by the step (2) by the step (1)
Implement LC-MS/MS analyses.The quality-control sample (QC) of basic, normal, high 3 kinds of concentration is prepared simultaneously, and quality-control sample number is no less than to work as and criticize
The 5% of total number of samples.S (+)-crystal methamphetamine, R (-)-crystal methamphetamine, S (+)-phenylpropyl alcohol are calculated according to the standard curve on the same day
The concentration of amine and R (-)-amphetamine.
In 86 parts of samples, 72 (83.7%) part urines only detect S (+)-crystal methamphetamine and S (+)-amphetamine, dense
It is respectively 0.33~89.73mg/L and 0.11~14.03mg/L to spend range, and intermediate concentration is respectively 7.01 and 1.18mg/L.It should
The concentration results of S (+)-crystal methamphetamine and S (+)-amphetamine are shown in Table 6 in 72 parts of positive urines.Wherein, S (+)-amphetamine/S
(+)-crystal methamphetamine ratio range is 0.04~1.27, average value 0.25, median 0.16.
The concentration results of S (+)-crystal methamphetamine and S (+)-amphetamine in the positive urine of 6 72 parts of table
In remaining 14 parts of urines, R the and S enantiomers of crystal methamphetamine and amphetamine have been detected simultaneously by, but still
Based on S- configurations.Referring to table 7, in this 14 parts positive urines, the concentration range of S (+)-crystal methamphetamine for 0.33~
The concentration range of 69.87mg/L, R (-)-crystal methamphetamine is LOQ~13.76mg/L, and is detected in five parts of samples wherein
Concentration to R (-)-crystal methamphetamine is less than LOQ；The ratio range of the R/S of crystal methamphetamine is 0.00~3.45；S (+)-benzene
The concentration range of propylamine is 0.15~14.94mg/L, R (-)-amphetamine is only detected in 4 parts of samples, except two parts of concentration are low
In outside LOQ, in addition two parts of concentration is respectively 0.06 and 0.29mg/L, and the ratio of the R/S of amphetamine is respectively 0.03 and 0.57.
Concentration and R/S ratios in the urine of R and S enantiomers that 7 14 parts of table detects crystal methamphetamine simultaneously
+a:It detects but less than LOQ (0.05mg/L)；N.D.b.:It does not detect.
Specific embodiments of the present invention are described in detail above, but it is intended only as example, it is of the invention and unlimited
It is formed on particular embodiments described above.To those skilled in the art, it is any to the equivalent modifications that carry out of the present invention and
It substitutes also all among scope of the invention.Therefore, the impartial conversion made without departing from the spirit and scope of the invention and
Modification, all should be contained within the scope of the invention.
Claims (9)
1. it is a kind of detect simultaneously R (-)-crystal methamphetamine in liquid biological sample, S (+)-crystal methamphetamine, R (-)-amphetamine and
The method of S (+)-amphetamine, which is characterized in that include the following steps：
(1) sample pre-treatments；
(2) LC-MS/MS is analyzed；
Wherein, the sample pre-treatments described in step (1) include：
At room temperature, liquid biological sample sample is taken, is placed in an EP pipes, and add in inner mark solution；Implement to be vortexed for the first time and shake
It swings, is then centrifuged for, supernatant is taken to be placed in another EP pipes, and add in methanol；Implement second of vortex oscillation, finally, sample introduction is extremely
LC-MS/MS systems.
It is 2. according to claim 1 while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-methylbenzene third
The method of amine, R (-)-amphetamine and S (+)-amphetamine, which is characterized in that the liquid biological sample is blood or urine.
It is 3. according to claim 1 while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-methylbenzene third
The method of amine, R (-)-amphetamine and S (+)-amphetamine, which is characterized in that the inner mark solution is molten for the methanol of 4- phenyibutylamines
Liquid.
It is 4. according to claim 3 while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-methylbenzene third
The method of amine, R (-)-amphetamine and S (+)-amphetamine, which is characterized in that a concentration of 0.015ug/mL of the inner mark solution.
It is 5. according to claim 1 while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-methylbenzene third
The method of amine, R (-)-amphetamine and S (+)-amphetamine, which is characterized in that the concrete operations of the sample pre-treatments include：
At room temperature, 20 μ L liquid biological sample samples are taken, are placed in an EP pipes, and add in 980 μ L inner mark solutions；Implement first
Secondary 15~20s of vortex oscillation, is then centrifuged for 1~2min, and 50 μ L supernatants is taken to be placed in another EP pipes, and adds in 450 μ L methanol；
Implement second of 5~10s of vortex oscillation, finally, sample introduction to LC-MS/MS systems.
It is 6. according to claim 5 while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-methylbenzene third
The method of amine, R (-)-amphetamine and S (+)-amphetamine, which is characterized in that the volume of the EP pipes is 2mL.
It is 7. according to claim 5 while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-methylbenzene third
The method of amine, R (-)-amphetamine and S (+)-amphetamine, which is characterized in that the rotating speed of the centrifugation is 10000~13000r/
min。
It is 8. according to claim 1 while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-methylbenzene third
The method of amine, R (-)-amphetamine and S (+)-amphetamine, which is characterized in that in the LC-MS/MS analyses described in step (2), chromatography
Condition includes：
Chromatographic column is Supelco Astec ChirobioticTMV2 chiral columns (2.1mm × 250mm, 5 μm)；Mobile phase：Methanol/
0.1% (v/v) glacial acetic acid/0.02% (v/v) ammonium hydroxide；Flow velocity：0.25mL/min；Sample size：20μL；Injector temperature：25℃；
Column temperature：25℃.
It is 9. according to claim 1 while detect R (-)-crystal methamphetamine in liquid biological sample, S (+)-methylbenzene third
The method of amine, R (-)-amphetamine and S (+)-amphetamine, which is characterized in that in the LC-MS/MS analyses described in step (2), mass spectrum
Condition includes：
Electron spray ionisation-positive ion mode (ESI+)；Collision gas (CAD)：7psi；Gas curtain gas (CUR)：30psi；Ion injection electricity
It presses (IS)：5500V；Ion source gas 1 (GS1)：35psi；Ion source gas 2 (GS2)：35psi；Ion source temperature (TEM)：500℃；
Scan mode：More reaction detections (MRM).
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110907542A (en) * | 2018-09-17 | 2020-03-24 | 复旦大学 | Method for detecting selegiline and metabolite thereof in saliva by liquid chromatography-mass spectrometry |
| CN114689757A (en) * | 2022-04-07 | 2022-07-01 | 中国计量科学研究院 | Preparation method and application of amphetamine-type drug hair standard substance |
-
2017
- 2017-12-15 CN CN201711354278.9A patent/CN108181385A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110907542A (en) * | 2018-09-17 | 2020-03-24 | 复旦大学 | Method for detecting selegiline and metabolite thereof in saliva by liquid chromatography-mass spectrometry |
| CN114689757A (en) * | 2022-04-07 | 2022-07-01 | 中国计量科学研究院 | Preparation method and application of amphetamine-type drug hair standard substance |
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