CN105929042A - Method for rapidly detecting tetramine in blood of toxic patients - Google Patents

Abstract

The invention discloses a method for rapidly detecting tetramine in the blood of toxic patients, and belongs to the technical field of chemical detection. The detection method has the advantages of high sensitivity and high efficiency. The detection method comprises the following steps: (1) preparing a standard solution; (2) preparing a test sample: adding 5 mL of blood into a 15 mL centrifuge tube, adding 5 mL of acetonitrile, performing vortex shaking for 2 minutes, carrying out centrifugation for 5 minutes under a rotation speed of 3200 r/min, and measuring the supernate by GC-MS.

Description

The quickly method of the Tetramine in detection toxic patient blood

Technical field

The invention discloses the detection method of a kind of Tetramine, specifically, be that one quickly detects toxonosis The method of the Tetramine in human blood, belongs to technical field of chemical detection.

Background technology

Tetramine (tetramine), chemical entitled tetramethylene-disulfo-tetramine, for quick-acting the killing mouse of severe toxicity Medicine, colourless, tasteless.It has severe toxicity to all homoiothermic animals, and the Ministry of Agriculture, Ministry of Public Health are once in 1982 Dispatch a joint document in year and forbid to use these compounds as raticide, but still have people to use and cause poisoning. For providing foundation the most reliably to the diagnoses and treatment of toxic patient, it is necessary to foundation one operation simple and fast, Qualitative accurate, highly sensitive detection method.Tetramine gas chromatography mass spectrometry method, as uniquely confirming method, does not has Unified national standard, tradition pretreatment process complexity is loaded down with trivial details, the most long, it usually needs several hours, difficult To adapt to the quickly poison inspection requirement of public health emergency.And often analyzing complex matrices such as: people Blood etc. time, owing to purifying not, impurity brings system into, makes instrument consume serious, even causes conclusion Misjudgment.

Summary of the invention

For the problems referred to above, it is an object of the invention to provide the side of Tetramine in a kind of toxic patient blood Method, the method sensitivity is good, and detection efficiency is high.

For solving the problems referred to above, the technical scheme that the present invention provides is such that

The method of the Tetramine in a kind of quick detection toxic patient blood, comprises the steps: successively

1) preparation of standard solution

Accurately weigh Tetramine standard substance 5.0mg, be placed in 100ml volumetric flask, dissolve and constant volume with acetonitrile, It is made into 50 μ g/ml contrast solutions；

2) preparation of test sample

Take blood 5ml and put in 15ml centrifuge tube, add 5ml acetonitrile, whirlpool concussion 2min；3200r/min Centrifugal 5min, takes gas chromatograph-mass spectrometer on supernatant and measures；

Described chromatographic condition is as follows:

Chromatographic column: HP-5MS, 20m × 0.18mm, thickness 0.18 μm；Carrier gas is high-purity helium；Sample introduction Amount: 30 μ l；During PTV sample introduction, shunting exhaust-valve controls the flow at 80ml/min, and sample introduction completes After, shunting exhaust-valve closes 1.0min；

Described Mass Spectrometry Conditions is as follows:

Electron impact ion source, electron energy 70eV；Scanning characteristic ion: m/z92, m/z 132, m/z 212, m/z 240；GC/MS interface temperature: 280 DEG C；Ion source temperature: 230 DEG C；Level Four bar temperature: 150 DEG C.

Described PTV temperature program(me) is as follows: 40 DEG C maintain 1.0min, rise to 230 DEG C with 230 DEG C/min speed Maintain 2min, rise to 300 DEG C with 230 DEG C/min speed and maintain 7min；Column flow rate: with constant current mode 0.5ml/min；Column oven: 90 DEG C of 2min, rises to 150 DEG C with 100 DEG C/min speed, with 50 DEG C/min speed Rate rises to 280 DEG C and maintains 4min.

Further, the method for the above-mentioned Tetramine in quickly detection toxic patient blood, described makings connection Agilent 7890A-5975C gas chromatograph-mass spectrometer is used with instrument.

Compared with prior art, the technical scheme that the application provides combines large volume sample injection, PTV sample introduction, logical Cross sim pattern and carry out Tetramine detection confirmation.The time-consuming about 20min of overall process is completed from sample pre-treatments to confirmation. The method tool operation simple and fast, qualitative accurate, sensitivity high, can apply to emergent inspection, for Patient's rescue work saves valuable time, reduces cost.

Accompanying drawing explanation

Fig. 1 is the mass spectrum of SIM type collection Tetramine；

Fig. 2 is the Tetramine standard solution collection of illustrative plates that liquid-liquid extraction method combines heavy caliber chromatographic column；

Fig. 3 is the blood extracting solution collection of illustrative plates that liquid-liquid extraction method combines heavy caliber chromatographic column；

Detailed description of the invention

Below in conjunction with detailed description of the invention, the claim of the present invention is described in further detail, but not Constitute any limitation of the invention, anyone limited number of time done in the claims in the present invention protection domain Amendment, still within the claims of the present invention.

The percentage concentration that the present invention relates to, is not particularly illustrated, and liquid is volumetric concentration, solute Finger mass concentration for solid.

Embodiment 1

1. materials and methods

1.1 instruments and reagent

Agilent 7890A-5975C gas chromatograph-mass spectrometer, is furnished with CTC Multifunctional sample injection device, PTV injection port； Superpure water machine (ELGAPURELAB ClassicUVF)；Ultrasonic cleaner；High speed centrifuge；C18 is solid Extract pillar (Tianjin Bonaaijieer Technology Co., Ltd) mutually

Acetonitrile (HPLC level, Fisher company of the U.S.), other reagent is analytical pure (traditional Chinese medicines group chemistry Reagent company limited).Experimental water is to filter through ELGAPURELAB ClassicUVF cleaning system Deionized water；Tetramine standard substance are purchased from State center for standard matter.

1.2 chromatographic condition

Chromatographic column: HP-5MS (20m × 0.18mm thickness 0.18 μm)；Carrier gas is high-purity helium；Sample size: 30μl；During PTV sample introduction (0.8min), shunting exhaust-valve controls the flow at 80ml/min, sample introduction After completing, shunting exhaust-valve closes (1.0min)；PTV temperature program(me) is as follows: 40 DEG C maintain 1.0min, Rise to 230 DEG C with 230 DEG C/min speed and maintain 2min, rise to 300 DEG C with 230 DEG C/min speed and maintain 7min. Column flow rate: with constant current mode 0.5ml/min.Column oven: 90 DEG C (2min), with 100 DEG C/min speed liter To 150 DEG C, rise to 280 DEG C with 50 DEG C/min speed and maintain 4min.

Use solvent purging function and the HP-5MS (20m × 0.18mm thickness 0.18 μm) of PTV injection port, Appearance time can be accelerated, thus reduces analysis time on the basis of ensureing sensitivity.

Shunting Splitless injecting samples mouth relative to common injector with common gas phase, general effective sample size is 1 μl；But the application have employed large volume sample injection (50ul) and PTV sample introduction pattern, for reaching to be not required to after extracting Extracting solution is concentrated, gets final product loading, and realize solvent purging at injection port end, after making object focus on Enter chromatographic column, be greatly saved the time of pre-treatment.

2.3 Mass Spectrometry Conditions

Electron bombardment (EI) ion source, electron energy 70eV；Scanning characteristic ion: m/z92, m/z 132, M/z 212, m/z 240 (mass spectrum is shown in Fig. 1)；GC/MS interface temperature: 280 DEG C；Ion source temperature: 230℃；Level Four bar temperature: 150 DEG C.

The preparation of 2.4 standard solution

Accurately weigh Tetramine standard substance 5.0mg, be placed in 100ml volumetric flask, dissolve and constant volume with acetonitrile, It is made into 50 μ g/ml contrast solutions.

The preparation of 2.5 test samples

Take blood 5ml and put in 15ml centrifuge tube, add 5ml acetonitrile, whirlpool concussion 2min；3200r/min Centrifugal 5min, takes gas chromatograph-mass spectrometer on supernatant and measures.

3 results

Being added in blank blood sample by Tetramine, " 2.5 " the joint method of employing processes, and measures Tetramine The response rate in blood, every kind of addition level determination 3 times, average and detect as reality, result It is shown in Table 1.

Result shows that Blood calldack rate is 80%～93%, and method accuracy rate is high.

Claims (2)

1. the method for the Tetramine in a quick detection toxic patient blood, it is characterised in that include successively Following step:

1) preparation of standard solution

Accurately weigh Tetramine standard substance 5.0mg, be placed in 100ml volumetric flask, dissolve and constant volume with acetonitrile, It is made into 50 μ g/ml contrast solutions；

2) preparation of test sample

Take blood 5ml and put in 15ml centrifuge tube, add 5ml acetonitrile, whirlpool concussion 2min；3200r/min Centrifugal 5min, takes gas chromatograph-mass spectrometer on supernatant and measures；

Described chromatographic condition is as follows:

Chromatographic column: HP-5MS, 20m × 0.18mm, thickness 0.18 μm；Carrier gas is high-purity helium；Sample introduction Amount: 30 μ l；During PTV sample introduction, shunting exhaust-valve controls the flow at 80ml/min, and sample introduction completes After, shunting exhaust-valve closes 1.0min；

Described Mass Spectrometry Conditions is as follows:

Electron impact ion source, electron energy 70eV；Scanning characteristic ion: m/z92, m/z 132, m/z 212, m/z 240；GC/MS interface temperature: 280 DEG C；Ion source temperature: 230 DEG C；Level Four bar temperature: 150 DEG C.

Described PTV temperature program(me) is as follows: 40 DEG C maintain 1.0min, rise to 230 DEG C with 230 DEG C/min speed Maintain 2min, rise to 300 DEG C with 230 DEG C/min speed and maintain 7min；Column flow rate: with constant current mode 0.5ml/min；Column oven: 90 DEG C of 2min, rises to 150 DEG C with 100 DEG C/min speed, with 50 DEG C/min speed Rate rises to 280 DEG C and maintains 4min.

The method of the Tetramine in quick detection toxic patient blood the most according to claim 1, it is special Levying and be, described gas chromatograph-mass spectrometer uses Agilent 7890A-5975C gas chromatograph-mass spectrometer.