CN107101864A - Method for preparing purified for the cocainehydrochloride standard substance of forensic science illicit drugs inspection - Google Patents

Method for preparing purified for the cocainehydrochloride standard substance of forensic science illicit drugs inspection Download PDF

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Publication number
CN107101864A
CN107101864A CN201710316042.XA CN201710316042A CN107101864A CN 107101864 A CN107101864 A CN 107101864A CN 201710316042 A CN201710316042 A CN 201710316042A CN 107101864 A CN107101864 A CN 107101864A
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China
Prior art keywords
cocaine
sample
cocainehydrochloride
standard substance
methanol
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Chinese (zh)
Inventor
郑珲
高利生
赵彦彪
张春水
赵阳
常颖
贺剑锋
翟晚枫
李彭
刘克林
钱振华
郑晓雨
杨虹贤
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Institute of Forensic Science Ministry of Public Security PRC
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Institute of Forensic Science Ministry of Public Security PRC
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/34—Purifying; Cleaning
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/74—Optical detectors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/86—Signal analysis
    • G01N30/8624—Detection of slopes or peaks; baseline correction
    • G01N30/8631—Peaks
    • G01N30/8634—Peak quality criteria
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/89—Inverse chromatography

Abstract

The present invention discloses the method for preparing purified of the cocainehydrochloride standard substance for forensic science illicit drugs inspection, comprises the following steps：(1) purity of cocaine in sample of cocaine is captured in detection, and selection cocaine mass fraction captures the raw material that sample of cocaine prepares cocainehydrochloride standard substance as purifying more than or equal to 60wt%；(2) cocainehydrochloride standard substance is prepared using high performance liquid chromatography.The present invention prepares the cocainehydrochloride that purification process obtains and confirmed through nuclear magnetic resonance, Liquid Chromatography-Tandem Mass Spectrometry combination, infrared spectrum analysis, and its purity confirms definite value through liquid chromatogram, gas-chromatography, and chromatogram is measured without response impurity；Required according to Developments of certified reference samples, its stability, uniformity, definite value, the estimation of overall uncertainty meet relevant regulations, reach expectation index.

Description

It is prepared by the purifying for the cocainehydrochloride standard substance of forensic science illicit drugs inspection Method
Technical field
The present invention relates to the preparation of forensic science drugs standard items.It is used for forensic science drugs more particularly, to one kind The method for preparing purified of the cocainehydrochloride standard substance of detection.
Background technology
At present, the drug issue being on the rise turns into global disaster.The body for spreading unchecked the directly harm people of drugs Heart health, and bring grave danger to economic growth and social progress.Therefore, forensic science illicit drugs inspection magnitude tracing body is set up System, improves drugs composition measurement technique, it is ensured that the reliability and comparativity of measurement result, sets up the shared and mutual of measurement data Recognize, accurately and reliably evidence is provided for court, it has also become countries in the world drugs appraisal organization question of common concern.
Drugs composition measurement technique and traceability guarantee are an organic wholes, are core measurement capabilities in section of court Learn the important embodiment of drugs ingredient amount fields of measurement.Standard substance is through, in this overall skeleton, being the carrier of value, being The key element of drugs ingredient amount traceability system, is the weight for ensureing measurement result accuracy over time and space and comparativity It is basic, it is to realize effectively i.e. accurate, comparable, the measurement that can trace to the source the basic assurance of measurement.
Being generally acknowledged in the world using what the companies such as Sigma produced mostly in the advanced illicit drugs inspection laboratory of the National Technicals such as America and Europe Standard substance, but China can only dependence on import standard substance, the few valency of amount is high, has many can't be to China's offer.Mesh Not only species is extremely limited " reference substance " used in preceding domestic drugs of abuse, and general lack of perfect Structural Identification, pure The corresponding technical indicator such as degree measure, uniformity and stability.These all carry out certain uncertainty to case detection band of being involved in drug traffic, Directly influence the accuracy of quantitative result.Moreover, the scarcity of domestic drugs standard substance, has become restriction China and realizes method Front yard science illicit drugs inspection chemical measurement methodological standardization, the major obstacle traced to the source and recognized each other of measurement result.Therefore, it is possible to prepare Go out the problem of having become China's drugs research field urgent need to resolve for the drugs standard substance of forensic science illicit drugs inspection.
The physicochemical property of cocaine：Also known as：Cocaine, english common name：Cocaine hydrochloride, chemistry Title：(-) -3 β-beta-methoxy formoxyl tropane of benzoyloxy -2, English name：2-Carbomethoxy-3-benzoyloxy Tropane hydrochloride, molecular formula：C17H21NO4HCl, molecular weight：339.8, CA registration numbers：53-21-4, structure Formula is：
Physicochemical property：Colourless, odorless monoclinic crystal.98 DEG C of fusing point.Water is practically insoluble in, is dissolved in general organic molten Agent.Taste is first bitter then numb.Colourless crystallization or white crystalline powder, more than 197 DEG C of fusing point.Hydrochloride is highly soluble in water, readily soluble In ethanol, chloroform is dissolved in, insoluble in ether.D-72 ° of optical activity [α] (c=2, water).It is stored in 2-8 DEG C.
The content of the invention
It is an object of the present invention to provide a kind of cocainehydrochloride standard substance for forensic science illicit drugs inspection Method for preparing purified.
To reach above-mentioned purpose, the present invention uses following technical proposals：
For the method for preparing purified of the cocainehydrochloride standard substance of forensic science illicit drugs inspection, comprise the following steps：
(1) purity of cocaine in sample of cocaine is captured in detection, and selection cocaine mass fraction is more than or equal to 60wt%'s captures the raw material that sample of cocaine prepares cocainehydrochloride standard substance as purifying；
(2) cocainehydrochloride standard substance is prepared using high performance liquid chromatography.
The method for preparing purified of the above-mentioned cocainehydrochloride standard substance for forensic science illicit drugs inspection, in step (1) In, comprise the following steps：
(1.1) preparation of sample solution：Weigh 10mg sample of cocaine to be dissolved in 10mL distilled water, be made into 1mg/mL cocker Because of sample solution, using preceding sample solution by 0.45 μm of filtering with microporous membrane；
(1.2) liquid phase chromatogram condition is determined：Chromatographic column is Shim-pack HRC-ODS posts, 250mm × 4.6mm I.D., 5 μm；Mobile phase is VMethanol：V0.05% trifluoroacetic acid/water=33:67, isocratic elution；Ultraviolet detection wavelength 230nm；Flow velocity 1.0mL/min；Post 35 DEG C of temperature；
(1.3) calculate the regression equation of standard curve and determine the range of linearity：Cocaine standard is diluted with Chromatographic Pure Methanol Storing solution, precision is configured to the cocaine reference substance solution that concentration is respectively 10,20,50,100,200,500,1000 μ g/mL, Determined by the reverse-phase chromatography condition in step (1.2), each concentration is repeated 3 times, with mean value calculation, record cocaine chromatographic peak Area, the sample introduction concentration using reference substance is abscissa, and chromatographic peak area value is mapped for ordinate, and calculates the recurrence of standard curve Equation；Concentration is mapped with peak area, the regression equation of standard curve is：
Y=3 × 107X+4781, R2=1.0；
Cocaine linear relationship in the range of 0.5-1000 μ g/mL is good；
(1.4) the sample of cocaine aqueous solution, replication 3 times, note are determined by the reverse-phase chromatography method analysis of step (1.2) Cocaine peak area is recorded, its average value is calculated, the cocaine content in material sample is calculated by peak area external standard method.
The method for preparing purified of the above-mentioned cocainehydrochloride standard substance for forensic science illicit drugs inspection, in step (2) In, comprise the following steps：
(2.1) preparation of sample solution：Sample of cocaine 3731.42mg is weighed in 50mL volumetric flasks, adds water and is settled to Scale；It is configured to the sample aqueous solution that cocaine content is 50mg/mL and isolates and purifies preparation for preparative high performance liquid chromatography Cocaine standard items；Sample solution ultrasonic dissolution 5min, uses preceding process 0.22um mixing membrane filtrations；
(2.2) liquid phase chromatogram condition is determined：Chromatographic column is that Shim-pack VP-ODS prepare post, 250mm × 20mm I.D., 5 μm；Mobile phase is by A phases and B phase compositions：A phases are that 0.05%TFA/ water, i.e. trifluoroacetic acid volume fraction are 0.05% Trifluoroacetic acid aqueous solution；B phases are methanol, and gradient condition is by taking B phases as an example：0-21min 30%, 21-25min 30%-50%, 25-30min 50%-65%, 30-38min 65%, 38-45min 30%；Or mobile phase is by A phases and B phase compositions, VMethanol： VWater=30:70, isocratic elution；Ultraviolet detection wavelength 230nm or 254nm；Flow velocity 8mL/min；The μ L of applied sample amount 600；Column temperature is room Temperature；
(2.3) cocaine is present in the form of hydrochloride in sample of cocaine, the cocaine after preparing liquid phase separation Hydrochloride is directly dissolved in methanol -0.05%TFA/ aqueous systems or methanol-water solution, remove cut in trifluoroacetic acid and/ Or methanol, cocainehydrochloride crystal can be obtained after freeze-drying.
Beneficial effects of the present invention are as follows：
The present invention is prepared in the cocaine crystal that purification process is obtained, according to high performance liquid chromatography areas of peak normalization method meter Calculate cocaine purity and be more than or equal to 99.1wt%.
The present invention prepares cocaine that purification process obtains through nuclear magnetic resonance, Liquid Chromatography-tandem Mass, infrared light Analysis of spectrum confirms that its purity confirms definite value through liquid chromatogram, gas-chromatography, and chromatogram is measured without response impurity；According to Developments of certified reference samples requirement, its stability, uniformity, definite value, the estimation of overall uncertainty meet relevant regulations, reach expection Index.
The present invention prepare purification process can be provided for judicial expertise department of China value accurately, the cocaine mark that can trace to the source Quasi- material, fills up China's forensic science field drugs standard substance blank, to improve analysis measurement quality, improves quantitative result The degree of accuracy, farthest ensures the validity of measurement result.Help to set up forensic science illicit drugs inspection magnitude tracing system, Domestic forensic science illicit drugs inspection chemical measurement methodological standardization is advantageously implemented, the reliable, effective and mutual of measurement result is realized Recognize.
Overcome prior art and prepare that the purity that the sample of cocaine that purification process obtains is present is low, stability is poor, uniform Property the defect such as poor, preparation process complexity there is provided one kind using high performance liquid chromatography separation method obtain high-purity, high stability, Rate of recovery height, the cocaine standard substance preparation method for being easy to large-scale production.
Brief description of the drawings
The embodiment to the present invention is described in further detail below in conjunction with the accompanying drawings.
The ultraviolet spectrogram of impurity 1 in Fig. 1-1 sample of cocaine；
The ultraviolet spectrogram of impurity 2 in Fig. 1-2 sample of cocaine；
The ultraviolet spectrogram of Fig. 1-3 cocaines；
Fig. 2-1 sample of cocaine HPLC chromatograms (VMethanol：V0.05%TFA/ water=40:60)；
Fig. 2-2 Fig. 2-1 partial enlarged drawing；
Fig. 3-1 sample of cocaine HPLC chromatograms (VMethanol：V0.05%TFA/ water=33:67)；
Fig. 3-2 Fig. 3-1 partial enlarged drawing；
Fig. 4 sample of cocaine prepares HPLC chromatogram (A：0.05%TFA/ water, B：Methanol, gradient condition (B) 0-21min 30%, 21-25min 30%-50%, 25-30min 50%-65%, 30-38min 65%, 38-45min 30%)；
Fig. 5 sample of cocaine prepares HPLC chromatogram, VMethanol：VWater=30:70；
HPLC chromatogram (the V of Fig. 6 cocaine cutsMethanol：V0.05%TFA/ water=33:67)；
The hydrogen spectrogram of Fig. 7 cocaines；The carbon spectrogram of Fig. 8 cocaines；
The infrared spectrum of Fig. 9 cocainehydrochlorides；The mass spectrogram of Figure 10 cocainehydrochlorides.
Embodiment
In order to illustrate more clearly of the present invention, the present invention is done further with reference to preferred embodiments and drawings It is bright.Similar part is indicated with identical reference in accompanying drawing.It will be appreciated by those skilled in the art that institute is specific below The content of description is illustrative and be not restrictive, and should not be limited the scope of the invention with this.
The present embodiment mainly uses the cocaine sample that case is captured, to isolating and purifying system using preparative liquid chromatography Screening is optimized in the experiment condition of standby cocainehydrochloride standard substance.
First, instrument, reagent and material
1.1 key instrument
Analytic type high performance liquid chromatograph (Japanese Shimadzu), including：LC-20AD high pressure pumps；SIL-10A auto injections Device；SPD-20A PDADs；CTO-20A column ovens.
Preparative high performance liquid chromatography instrument (Agilent), including：G1361A high pressure pumps；G2260A automatic samplers； G1315D PDADs；G1364B automatic fraction collectors.
Preparative high performance liquid chromatography instrument (Japanese Shimadzu), including：LC-8A high pressure pumps；SIL-10AP auto injections Device；SPD-M20A PDADs；FRC-10A automatic fraction collectors；CBM-20A system controllers.BUCHI rotates Evaporator (Japanese BUCHI companies)；KQ3200 types ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.)；Flying pigeon board TDL- 40B desk centrifuges (Anting Scientific Instrument Factory, Shanghai)；XS105Dual Range electronic balances (Switzerland METTLER TOLEDO Company).
1.2 main agents and material
Methanol, trifluoroacetic acid (chromatographically pure, Chinese lark prestige company), ultra-pure water is (through Millipore ultrapure water production systems Purification, French Millipore companies).Cocaine 1mg/mL standard liquids (Chinese lark prestige company).Sample of cocaine (white Powder) captured and be applied in the present embodiment by case.
2nd, detection captures in sample of cocaine the purity of cocaine and prepares cocaine using sample of cocaine purifying is captured Standard substance
The preparation of 2.1 sample solutions
Analytic type：Weigh 10mg sample of cocaine to be dissolved in 10mL distilled water, be made into 1mg/mL sample of cocaine solution, make With preceding sample solution by 0.45 μm of filtering with microporous membrane.
Preparative：Sample of cocaine 3731.42mg (equivalent to pure cocaine 2500mg) is weighed in 50mL volumetric flasks, plus Water is settled to scale.The sample aqueous solution that cocaine content is 50mg/mL is configured to separate for preparative high performance liquid chromatography Prepare cocaine standard items.Sample solution ultrasonic dissolution 5min, uses preceding process 0.22um mixing membrane filtrations.
2.2 liquid phase chromatogram condition
2.2.1 RPLC (RP-HPLC) analysis method:
Chromatographic column is Shim-pack HRC-ODS posts (250mm × 4.6mm I.D., 5 μm)；Mobile phase is VMethanol： V0.05% trifluoroacetic acid/water=33:67 (0.05%TFA/ water refers to the trifluoroacetic acid aqueous solution that trifluoroacetic acid volume fraction is 0.05%), Isocratic elution；Ultraviolet detection wavelength 230nm；Flow velocity 1.0mL/min；35 DEG C of column temperature.
2.2.2 RPLC (RP-HPLC) preparation method:
Chromatographic column is that Shim-pack VP-ODS prepare post (250mm × 20mm I.D., 5 μm)；Flow combined one：Stream Move by A phases and B phase compositions：A phases are that the trifluoroacetic acid that 0.05%TFA/ water, i.e. trifluoroacetic acid volume fraction are 0.05% is water-soluble Liquid；B phases are methanol, and gradient condition is illustrated by taking B phases as an example：0-21min 30%, 21-25min 30%-50%, 25- 30min 50%-65%, 30-38min 65%, 38-45min 30%, i.e.,：B phase volumes are 30% and 21 points of holding when initial Clock, increases to 50% at 25 minutes, increase to 65% at 30 minutes and kept for 8 minutes, and 30% and holding 7 are reduced at 38 minutes Minute.
Flow combined two：VMethanol：VWater=30:70, isocratic elution.
Ultraviolet detection wavelength 230nm or 254nm；Flow velocity 8mL/min；The μ L of applied sample amount 600；Column temperature is room temperature.
The optimization of 2.3 efficient liquid phase chromatographic analysis conditions
2.3.1 the selection of chromatographic column
The reverse-phase chromatographic column that this experiment is used is Shim-pack HRC-ODS posts (250mm × 4.6mm I.D., 5 μm).
2.3.2 the selection of Detection wavelength
Analyzed by HPLC-DAD, in the range of 190-500nm, cocaine and its major impurity are equal for 230nm in wavelength There is absorption, as shown in Fig. 1-1 to Fig. 1-3, so selection Detection wavelength is 230nm.
2.3.3 the selection of flow visualizing
By the cocaine aqueous sample of preparation, HPLC analyses are carried out, respectively from VMethanol：V0.05%TFA/ water=33:67 and 40: The flow visualizing of 60 two kinds of different proportions, is separated in the case of other conditions identical to sample.Work as mobile phase ratio For VMethanol：V0.05%TFA/ water=40：When 60, cocaine is not completely separated (Fig. 2-1 and Fig. 2-2) with adjacent component；When mobile phase ratio Example is VMethanol：V0.05%TFA/ water=33：When 67, cocaine can reach baseline separation (Fig. 3-1 and Fig. 3-2) with adjacent component.So V is selected in this experimentMethanol：V0.05%TFA/ water=33：67 are used as the flow visualizing for analyzing sample of cocaine.
2.4 standard curves and linear relationship
With chromatogram methanol dilution cocaine standard reserving solution, precision be configured to concentration be respectively 10,20,50,100,200, 500th, 1000 μ g/mL cocaine reference substance solution, is determined, each concentration repeats 3 by step 2.2.1 reverse-phase chromatography condition It is secondary, with mean value calculation, cocaine chromatographic peak area is recorded, with the sample introduction concentration (μ g/mL) of reference substance for abscissa, chromatographic peak Area value is mapped for ordinate, and calculates the regression equation of standard curve.
The concentration and peak area of the cocaine of table 1
Concentration is mapped with peak area, the regression equation of standard curve is：
Y=3 × 107X+4781, R2=1.0 ... ... ... ...
Show that cocaine linear relationship in the range of 0.5-1000 μ g/mL is good.
The measure of cocaine content in 2.5 sample solutions
The sample of cocaine aqueous solution is determined by 2.2.1 reverse-phase chromatography method analysis, replication 3 times records cocaine Peak area, calculates its average value, and the cocaine content in material sample is calculated by peak area external standard method.
The measurement result of cocaine content in the sample of table 2
As shown in Table 12, calculated by peak area external standard method by calibration curve equation and obtain containing for cocaine in material sample Measure as 67.04wt%.
The optimization of 2.6 high performance liquid chromatography preparation conditions
The cocaine component in sample of cocaine is separated using methanol-TFA/ aqueous systems identical with analysis liquid phase, although Cocaine can reach good separation with the other components in sample in analysis liquid phase, but cocaine is simultaneously in liquid phase is prepared It can not reach and efficiently separate with other components, and cocaine is because content is higher and conditions of streaking occurs, attempts reduction The ratio of methanol, finally using methanol-TFA/ aqueous systems, condition of gradient elution, cocaine has reached preferably with other components Separating effect, prepares liquid chromatography(LC figure as shown in Figure 4.
Due to preparing cocaine component using methanol-TFA/ aqueous systems, needed the trifluoro in cut in the later stage of preparation Acetic acid is removed, and is operationally added with organic solvent extraction and acidified process under alkalescence condition, to simplify preparation method, The chance that impurity is introduced in cost, reduction mark thing preparation process is reduced, the present embodiment prepares cocaine to methanol-water solution again and entered Conditional filtering is gone, finally using VMethanol：VWater=30:70 can reach preferable separating effect.Preparative liquid chromatography figure such as Fig. 5 It is shown.
2.7 cocaines are further purified
Cocaine is present in the form of hydrochloride in sample of cocaine, the cocaine hydrochloride after preparing liquid phase separation It is directly dissolved in methanol-water solution, therefore in order to obtain cocainehydrochloride crystal, further purification process need to be passed through.Pass through Revolving removes the methanol in cut, and cocainehydrochloride crystal can be obtained after freeze-drying.Its HPLC chromatogram such as Fig. 6 institutes Show.Analyzed through HPLC, areas of peak normalization method determines percentage composition, cocaine content 99.874wt%.
3rd, the structural identification of cocaine standard substance
3.1 nuclear magnetic resonance spectroscopy
Structural identification has been carried out to cocainehydrochloride using 600M nuclear-magnetisms (Bruker AV600).Sample is dissolved in deuterated first Alcohol.As a result shown in below figure 7 and Fig. 8.
Solvent：D2O
3.2 infrared spectrum analysis
The red of cocainehydrochloride is determined on AVATAR 330FT-IR infrared spectrometers (Thermo Nicolet companies) External spectrum, (as shown in Figure 9) as a result consistent with standard spectrogram.
3.3 Liquid Chromatography-tandem Mass are analyzed
The mass spectrum of cocainehydrochloride is determined on the instrument of Agilent LC-MS/MS 6410.Under ESI positive ion modes, Broken voltage 80V, impact energy 8eV, the mass spectrogram of sample is as shown in Figure 10.
Following message can be obtained from mass spectrogram：
(1) molecular ion peak [M+H]+m/z304.30, differed with the relative molecular mass 303.353g/mol of cocaine 1, thus the detection molecular weight of sample and being consistent for cocaine.
(2) molecular ion peak m/z303 differs 121 with adjacent peak m/z182, and the fragment of loss is probably benzoate anion.
The definite value result of 3.4 cocaine standard substances
Analyzed through HPLC and GC methods definite value, the purity of cocainehydrochloride standard substance is 99.48wt%, is extended not true Fixed degree is 0.06% (k=2).Have good uniformity, stability at least more than 1 year.
Obviously, the above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not pair The restriction of embodiments of the present invention, for those of ordinary skill in the field, may be used also on the basis of the above description To make other changes in different forms, all embodiments can not be exhaustive here, it is every to belong to this hair Row of the obvious changes or variations that bright technical scheme is extended out still in protection scope of the present invention.

Claims (3)

1. the method for preparing purified of the cocainehydrochloride standard substance for forensic science illicit drugs inspection, it is characterised in that including Following steps：
(1) purity of cocaine in sample of cocaine is captured in detection, and selection cocaine mass fraction is more than or equal to 60wt%'s Capture the raw material that sample of cocaine prepares cocaine standard substance as purifying；
(2) cocainehydrochloride standard substance is prepared using high performance liquid chromatography.
2. the purifying preparation side of the cocainehydrochloride standard substance according to claim 1 for forensic science illicit drugs inspection Method, it is characterised in that in step (1), comprise the following steps：
(1.1) preparation of sample solution：Weigh 10mg sample of cocaine to be dissolved in 10mL distilled water, be made into 1mg/mL cocaine samples Product solution, using preceding sample solution by 0.45 μm of filtering with microporous membrane；
(1.2) liquid phase chromatogram condition is determined：Chromatographic column is Shim-pack HRC-ODS posts, 250mm × 4.6mm I.D., 5 μm； Mobile phase is VMethanol：V0.05% trifluoroacetic acid/water=33:67, isocratic elution；Ultraviolet detection wavelength 230nm；Flow velocity 1.0mL/min；Column temperature 35 ℃；
(1.3) calculate the regression equation of standard curve and determine the range of linearity：Cocaine standard inventory is diluted with Chromatographic Pure Methanol Liquid, precision is configured to the cocaine reference substance solution that concentration is respectively 10,20,50,100,200,500,1000 μ g/mL, by step Suddenly the reverse-phase chromatography condition in (1.2) is determined, and each concentration is repeated 3 times, with mean value calculation, record cocaine chromatographic peak face Product, the sample introduction concentration using reference substance is abscissa, and chromatographic peak area value is mapped for ordinate, and calculates the recurrence side of standard curve Journey；Concentration is mapped with peak area, the regression equation of standard curve is：
Y=3 × 107X+4781, R2=1.0；
Cocaine linear relationship in the range of 0.5-1000 μ g/mL is good；
(1.4) by the reverse-phase chromatography method analysis measure sample of cocaine aqueous solution of step (1.2), replication 3 times, record can Cacaine peak area, calculates its average value, and the cocaine content in material sample is calculated by peak area external standard method.
3. the purifying preparation side of the cocainehydrochloride standard substance according to claim 1 for forensic science illicit drugs inspection Method, it is characterised in that in step (2), comprise the following steps：
(2.1) preparation of sample solution：Sample of cocaine 3731.42mg is weighed in 50mL volumetric flasks, adds water and is settled to scale； The sample aqueous solution that cocaine content is 50mg/mL is configured to isolate and purify for preparative high performance liquid chromatography and prepare cocaine Standard items；Sample solution ultrasonic dissolution 5min, uses preceding process 0.22um mixing membrane filtrations；
(2.2) liquid phase chromatogram condition is determined：Chromatographic column is that Shim-pack VP-ODS prepare post, 250mm × 20mm I.D., 5 μ m；Mobile phase is by A phases and B phase compositions：A phases are the trifluoroacetic acid that 0.05%TFA/ water, i.e. trifluoroacetic acid volume fraction are 0.05% The aqueous solution；B phases are methanol, and gradient condition is by taking B phases as an example：0-21min 30%, 21-25min 30%-50%, 25-30min 50%-65%, 30-38min 65%, 38-45min 30%；Or mobile phase is by A phases and B phase compositions, VMethanol：VWater=30:70, Isocratic elution；Ultraviolet detection wavelength 230nm or 254nm；Flow velocity 8mL/min；The μ L of applied sample amount 600；Column temperature is room temperature；
(2.3) cocaine is present in the form of hydrochloride in sample of cocaine, the cocaine hydrochloric acid after preparing liquid phase separation Salt is directly dissolved in methanol -0.05%TFA/ aqueous systems or methanol-water solution, removes trifluoroacetic acid and/or first in cut Alcohol, can obtain cocainehydrochloride crystal after freeze-drying.
CN201710316042.XA 2017-05-08 2017-05-08 Method for preparing purified for the cocainehydrochloride standard substance of forensic science illicit drugs inspection Pending CN107101864A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110095458A (en) * 2019-06-05 2019-08-06 公安部禁毒情报技术中心 The preparation method of crystal methamphetamine urine detection kit quality control test panel

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101750458A (en) * 2009-11-05 2010-06-23 香港科技大学 Method for detecting drug residues in human body
CN102759476A (en) * 2012-07-20 2012-10-31 公安部禁毒情报技术中心 Preparation method of cocaine hydrochloride standard substance

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101750458A (en) * 2009-11-05 2010-06-23 香港科技大学 Method for detecting drug residues in human body
CN102759476A (en) * 2012-07-20 2012-10-31 公安部禁毒情报技术中心 Preparation method of cocaine hydrochloride standard substance

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
李雅宁: "毒品2C-E杂质成分分析以及毒品标准品的制备方法研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 *
梁宇华: "可卡因的定性定量分析", 《刑事技术》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110095458A (en) * 2019-06-05 2019-08-06 公安部禁毒情报技术中心 The preparation method of crystal methamphetamine urine detection kit quality control test panel

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