CN105223263B - A kind of detection platform and detection method for being used to measure trace element in biological sample - Google Patents
A kind of detection platform and detection method for being used to measure trace element in biological sample Download PDFInfo
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- CN105223263B CN105223263B CN201410240392.9A CN201410240392A CN105223263B CN 105223263 B CN105223263 B CN 105223263B CN 201410240392 A CN201410240392 A CN 201410240392A CN 105223263 B CN105223263 B CN 105223263B
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Abstract
The present invention relates to a kind of detection platform and detection method for being used to measure trace element in biological sample, the detection platform includes：By the Frozen Biological to solid refrigeration system；The laser ablation system of laser ablation is carried out to being refrigerated to solid biological sample；And the aerosol of the biological sample for laser ablation to be obtained is loaded into ionization in plasma and detects the inductivity coupled plasma mass spectrometry system of the Determination of trace element contents in the biological sample.
Description
Technical field
The invention belongs to the Element detection field of biomaterial, and in particular to one kind is adapted to measure using LA-ICP-MS methods
The detection platform and detection method of trace element in biological sample.
Background technology
Inductivity coupled plasma mass spectrometry technology (ICP-MS), which comes out so far, to be had 25 years.In the time of more than 20 years,
It develops into a kind of analytical technology for being widely used and being greatly prized by rapidly.The existing thousands of ICP-MS in the whole world are as conventional
Analysis means, are equipped in the laboratory of different research fields.With the development and perfection of instrument itself, it is in trace, ultra trace
Advantageous ability is shown in elemental analysis.
The method of trace element using classical wet digestion, micro-wave digestion or is in traditional analysis biological sample
High-pressure digestion tank is cleared up, its principle is that the strand of biological sample is carried out using the strong oxidizing property of nitric acid or hydrogen peroxide
Destroy, it is dissolved into solution, enter back into ICP-MS and be detected.Such method usually has higher want to the quantity of sample
Ask, general sample of clearing up is at least in 0.1g or so.Also, unnecessary impurity element can be also introduced in digestion process.
Laser ablation inductively coupled plasma-mass spectrometry technology (LA-ICP-MS) is that a kind of solid sample is introduced directly into skill
Art, its basic principle are that laser microbeam is focused on sample surfaces to be allowed to corrosion vaporization, are carried the particulate etched by carrier gas
Enter and ionized into plasma, then analyze and detect through mass spectrometer system.As a kind of introducing suitable for polytype solid sample
Method, with disturbing less, the inductivity coupled plasma mass spectrometry (ICP-MS) of high sensitivity be combined, opened up analytical technique of mass spectrum
Frontier.
The LA-ICP-MS technologies its have the following advantages that：(1) it is in situ, in real time, quickly analyze advantage；High sensitivity, sky
Between resolution ratio it is preferable the characteristics of.(2) analyzable sample scope is very wide, and almost all of sample can be by laser institute
Degrade.(3) solid sample can not have to decompose directly to measure, and improve sample introduction efficiency.(4) analysis of different modes can be taken,
Such as micro-zone analysis, global analysis or probe analysis.(5) to sample almost without requiring.Above-mentioned solid sample is introduced directly into
Technology (i.e. solid injection) reduces cumbersome sample preparation procedure, and possibility that is not only time saving and can reducing pollution, avoids
Solution prepare in dilution effect, to reduce detection limit it is favourable, and the Dry aerosol for introducing plasma cause mass spectrum interference compared with
Solwution method is less.
But conventional LA-ICP-MS methods in the analysis process more uses solid standard specimen, or school is carried out using solution standard specimen
Just, solid standard specimen uses the technique such as pressed powder, molten sintering in preparation process, and for biological sample then without good
Standard manufacturing method.Due to the repeatability during laser ablation, the uniformity of solid sample, solid sample in itself physical
Matter and surface state, the fractionation effect etc. of element drastically influence precision of analysis and precision.Therefore, how to pass through
The content of trace element in LA-ICP-MS method Accurate Determining biological samples, for one of the research hotspot in the field.
The content of the invention
It is contemplated that overcome detecting step in conventional wet decomposition means cumbersome and existing LA-ICP-MS methods in mark
The problem of quasi- sample preparation, be used to measure the detection platform of trace element and detection side in biological sample the present invention provides a kind of
Method, so as to quickly and accurately measure the trace element in biological sample.
An aspect of of the present present invention provides a kind of detection platform for being used to measure trace element in biological sample, the detection
Platform includes：By the Frozen Biological to solid refrigeration system；Laser stripping is carried out to being refrigerated to solid biological sample
The laser ablation system of erosion；And the aerosol of the biological sample for laser ablation to be obtained is loaded into plasma and ionizes
Change and detect the inductivity coupled plasma mass spectrometry system of the Determination of trace element contents in the biological sample.
Detection platform provided by the invention, can obtain being adapted in LA- effectively by biological sample snap frozen to solid-state
The solid standard specimen used in ICP-MS methods, and without any chemical reagents are introduced in detection process, environment-friendly high-efficiency, is conducive to
The development of subsequent detection process.The detection platform of the present invention is simple in structure, easy to use, and refrigeration is rapid, can quickly measure biology
The content of trace element in sample.
In the present invention, the refrigeration system includes：Store the sample cell of biological sample；There is opening and appearance at top
Receive the housing of at least one sample cell；The laser that the opening can be sealed and allow the laser ablation system to send is worn
Saturating lid；For the refrigeration unit for freezing biological sample and being kept the temperature；And for measuring the temperature of the biological sample
Temperature measuring unit.
Housing provided by the invention can accommodate multiple sample cells at the same time, can be at the same time via refrigeration unit by multiple samples
Freezed and kept the temperature, be conducive to subsequent sample detection, the rapid development of calibration steps, improve the speed of detection.Pass through lid
Body can be fully sealed with housing, closed cavity, and the laser-light transparent lid that laser ablation system is sent be formed, to being contained in
The solid biological sample that is refrigerated in housing carries out laser ablation, so as to obtain what is be subsequently detected with LA-ICP-MS methods
Aerosol.Also, by setting temperature measuring unit to be conducive to control refrigeration unit to keep the temperature the biological sample after freezing.
In the present invention, the refrigeration unit possesses for providing refrigerant into the housing to freeze biological sample
Refrigeration pipeline and refrigerant in the refrigeration pipeline according to the temperature control that the temperature measuring unit detects flow velocity
Flow control member.
Using refrigerant is provided into housing to freeze the refrigeration pipeline of biological sample and according to above-mentioned survey in the present invention
The flow control member of the flow velocity of refrigerant in the temperature controlled refrigerating pipeline that warm unit detects, effectively to freeze
Biological sample is simultaneously kept the temperature.Preferably, refrigerant can be nitrogen, so as to freeze more quickly.
In the present invention, the refrigeration pipeline include being respectively arranged on refrigerant-medium inlet portion on the side wall of the housing and
Refrigerant outlet portion and it is connected between the refrigerant-medium inlet portion and refrigerant outlet portion and is arranged at the shell
The internal tube body for being used to load the sample cell；And the flow control member is arranged on the refrigerant-medium inlet portion.
According to the present invention, the refrigerant as refrigeration source flowed into from refrigerant-medium inlet portion is flowed through in housing
Flowed out after tube body from refrigerant outlet portion.So as to by the refrigerant to circulate in tube body effectively to being placed in the pipe
The biological sample in sample cell on body is freezed.Also, by the flow control member for being arranged at refrigerant-medium inlet portion
The flow velocity that the temperature convection that can be effectively measured based on above-mentioned temperature measuring unit enters the refrigerant in housing is controlled, with favourable
Biological sample after to freezing is kept the temperature.Preferably, above-mentioned tubular body can be formed as M types, so as to more uniformly make
It is cold.
In the present invention, the temperature measuring unit includes the temperature measurer being connected with the sample cell.
According to the present invention, the temperature of the biological sample in sample cell can be effectively detected by the temperature measurer being connected with sample cell
Degree, is advantageously implemented heat insulation function.Preferably, which can be the temperature detecting resistance being connected with sample cell.
In the present invention, the inductivity coupled plasma mass spectrometry system possesses for the biological sample in the housing
The icp ms that the gas-carrier pipeline of carrier gas is provided and is connected with the gas-carrier pipeline.
According to the present invention, carrier gas can be provided to the biological sample in housing by gas-carrier pipeline, so that laser ablation obtains
The aerosol of biological sample ionization in plasma is loaded into by carrier gas, to carry out subsequent detection.
In the present invention, the gas-carrier pipeline includes carrier gas inlet portion and the carrier gas being respectively arranged on the side wall of the housing
Outlet portion, the carrier gas inlet portion are connected with carrier gas supply source, and the carrier gas outlet portion and the inductively coupled plasma
Mass spectrograph is connected.
In the present invention, gas-carrier pipeline includes carrier gas inlet portion and the carrier gas outlet portion being respectively arranged on the side wall of housing, institute
Carrier gas inlet portion is connected with carrier gas supply source to provide carrier gas, carrier gas outlet portion and inductivity coupled plasma mass spectrometry into housing
Instrument, which is connected, provides the aerosol for the biological sample being loaded into by carrier gas with latter.I.e., after laser ablation, lead into housing
Enter carrier gas, the aerosol degraded is brought into icp ms by carrier gas, to carry out subsequent detection.Into one
Step simplifies detection method, it is ensured that the accuracy of testing result.Preferably, which for example can be helium, argon gas etc
Inert gas.It is highly preferred that the carrier gas can be the mixed gas of helium and argon gas.
Present invention also offers a kind of detection method using above-mentioned detection platform, including：
(1) solid biological sample and standard solution will be refrigerated to via refrigeration system, will be carried out by laser ablation system
Laser ablation processing, forms the aerosol of biological sample and standard solution respectively；
(2) by inductivity coupled plasma mass spectrometry system by the aerosol obtained in step (1) be loaded into etc. from
It is detected after being ionized in daughter, the ion-intensity values of the biological sample and standard solution is directly obtained, according to the standard
The ionic strength of solution is worth to the calibration curve on ion-intensity values-concentration of the standard solution, and according to from described
The relational expression that calibration curve obtains converts to obtain the trace member in the biological sample with the ion-intensity values of the biological sample
The content of element.
In the method for the invention, can be rapidly molten by biological sample and standard due to the use of above-mentioned detection platform
Liquid is refrigerated to solid-state, recycles laser ablation system and inductivity coupled plasma mass spectrometry system to standard solution and biological sample
Tested, quantitative analysis is carried out to biological sample using standard solution as quantitative basis.Due to being in unified low-temperature condition
Standard specimen and sample are analyzed when similarly hereinafter, it can be ensured that the stability of data, the accuracy for improving data, so as to quickly and accurately obtain
The measured value of trace element into biological sample, and overcome that detecting step in conventional wet decomposition means is cumbersome and existing LA-
The problem of prepared by the standard sample in ICP-MS methods.
In the present invention, in the step (1), the laser ablation system degrades pattern using line scanning, and described
The wavelength for the laser that laser ablation system is sent is 213mm, pulse frequency 20Hz, aperture is degraded for 15~200 μm, when degrading
Between be 15~200 seconds.
By the selection of the parameter of above-mentioned laser ablation system, laser ablation can be effectively passed through and obtain biomaterial
Aerosol.
In the present invention, in the step (2), the inductivity coupled plasma mass spectrometry system is divided with jumping the time at peak
Distinguish type collection data, and in the inductivity coupled plasma mass spectrometry system, RF power is 1000w, sampling depth is
9mm, working gas are argon gas, and plasma flow amount is 15.0L/ minute, and carrier gas flux is 1.4L/ minutes, during data acquisition
Between be 120 seconds, the time of integration be 60 seconds.
, can be effectively to by laser ablation by the selection of the parameter of above-mentioned inductivity coupled plasma mass spectrometry system
Obtained from the aerosol of biomaterial be detected.
Beneficial effects of the present invention：
By biological sample and standard solution rapid condensation into solid-state under the refrigeration of refrigeration source, thus to sample into
Row fast and accurately quantitative analysis.Refrigerant system configurations are simple, easy to use, and refrigeration is rapid, and low temperature is kept constant.And put
Multigroup sample and standard solution can be analyzed within the same time by putting multi-disc sample cell.Unified low-temperature condition similarly hereinafter when analyze
Standard specimen and sample, it is ensured that the stability of data, the accuracy for improving data.The method solves traditional biological sample of resolution method analysis
The problem of complex for operation step in product, and any chemical reagents are not introduced, more environment-friendly high-efficiency, and solve sample preparation
When reagent blank it is excessive the problem of, improve detection sensitivity.The method also solves to be difficult in conventional LA-ICP-MS analyses
The problem of preparing solid standard specimen, prepares compared to traditional LA solids standard specimen, and solution standard specimen prepares more simple, quick, accurate.Mark
The uniformity of sample has also obtained maximum guarantee.And various carrier gas can be filled with the housing, reduce polyatom particle
Interference.
Brief description of the drawings
Fig. 1 shows the detection platform for being used to measure trace element in biological sample of an embodiment according to the present invention
Refrigeration system schematic diagram；
Fig. 2 is the schematic diagram of the sample cell used in the refrigeration system shown in Fig. 1；
Fig. 3 is the schematic diagram of the refrigeration unit used in the refrigeration system shown in Fig. 1；
Fig. 4 is used to measure the embodiment that the detection platform of trace element in biological sample is detected to be using the present invention
In obtained calibration curve.
Embodiment
Further illustrate the present invention below in conjunction with attached drawing and following embodiments, it should be appreciated that following embodiments and/
Or attached drawing is merely to illustrate the present invention, it is not intended to limit the present invention.
A kind of detection platform for being used to measure trace element in biological sample is provided according to an aspect of the present invention, it is described
Detection platform includes：By the Frozen Biological to solid refrigeration system；Swash to being refrigerated to solid biological sample
The laser ablation system that light degrades；And the aerosol of the biological sample for laser ablation to be obtained is loaded into plasma
Ionization and the inductivity coupled plasma mass spectrometry system for detecting the Determination of trace element contents in the biological sample.
Detection platform provided by the invention, can obtain being adapted in LA- effectively by biological sample snap frozen to solid-state
The solid standard specimen used in ICP-MS methods, and without any chemical reagents are introduced in detection process, environment-friendly high-efficiency, is conducive to
The development of subsequent detection process.The detection platform of the present invention is simple in structure, easy to use, and refrigeration is rapid, can quickly measure biology
The content of trace element in sample.
Fig. 1 shows the detection platform for being used to measure trace element in biological sample of an embodiment according to the present invention
Refrigeration system schematic diagram；Fig. 2 is the schematic diagram of the sample cell used in the refrigeration system shown in Fig. 1.Such as Fig. 1 and Fig. 2 institutes
Show, which includes：Store the sample cell 6 of biological sample；There is opening at top and accommodate at least one sample cell 6
Housing 2；The lid 1 for the laser penetration that the opening and permission laser ablation system are sent can be sealed；For freezing biology
Sample and the refrigeration unit kept the temperature；And the temperature measuring unit of the temperature for measuring the biological sample.The laser ablation
System can be laser ablation system commonly used in the prior art, omit the detailed description of its structure herein.
Multiple sample cells 6 can be accommodated at the same time in above-mentioned housing 2, can at the same time via refrigeration unit by multiple samples into
Row is freezed and kept the temperature.It can be fully sealed by lid 1 with housing 2, form closed cavity, such as in the embodiment shown in Fig. 1
In be formed as cube-shaped closed cavity.And the laser-light transparent lid 1 that laser ablation system is sent, to being contained in housing
The solid biological sample that is refrigerated in 2 carries out laser ablation, so that the gas for obtaining subsequently being detected with LA-ICP-MS methods is molten
Glue.Also, by setting temperature measuring unit to be conducive to control refrigeration unit to keep the temperature the biological sample after freezing.
Specifically, above-mentioned housing 2 is prepared using low temperature resistant, high intensity stainless steel material, is resistant to -50 DEG C or so
Low temperature and indeformable.The material of above-mentioned lid 1 is the quartz glass of penetrating bubble-free, which allows laser energy
Decay through without rapid, and focus on biological material specimens surface.
In addition, the material of above-mentioned sample cell 6 can be the aluminium nitride ceramics of polishing both surfaces, and in the one side of the sample cell 6
Smooth concave groove body 7 is processed, for placing biological sample or standard solution.Aluminium nitride ceramics thermal conductivity factor is big, resistance to
Strong acid and strong base is not easily susceptible to sample corrosion, is unlikely to deform fragmentation at low temperature.Can be quick and uniform by the sample cell of its making
Heat is conducted, whole groove body can be made uniformly to cool down and can preferably keep low-temperature condition.And its can carry it is various acid and
The sample of alkalescence, and can repeatedly use, embody it compared to more other similar materials such as stainless steel and aluminium oxide
Excellent performance.
In addition, above-mentioned refrigeration unit possesses for providing refrigerant into housing 2 to freeze the refrigerator pipes of biological sample
The flow control member of the flow velocity of road and the refrigerant in the temperature controlled refrigerating pipeline detected according to above-mentioned temperature measuring unit,
Effectively to freeze biological sample and to be kept the temperature.In the embodiment of the present invention, which can be nitrogen,
So as to freeze more quickly.
Specifically, Fig. 3 is the schematic diagram of the refrigeration unit used in the refrigeration system of the embodiment shown in Fig. 1.At this
In embodiment, refrigeration source is used as using nitrogen.As shown in figures 1 and 3, above-mentioned refrigeration pipeline includes being respectively arranged on housing 2
The nitrogen input pipe 4 as refrigerant-medium inlet portion on side wall and the nitrogen efferent duct 4 ' as refrigerant outlet portion, with
And it is connected between nitrogen input pipe 4 and nitrogen efferent duct 4 ' and is arranged at the tube body for being used to load sample cell 6 in housing 2
10.In this embodiment, which can for example be made of stainless steel tube.And flow control member 5 is arranged on the nitrogen
At input pipe 4.In this embodiment, which for example can be flow control valve.
As indicated by the arrows in fig. 3, the refrigerant stream of such as nitrogen as refrigeration source flowed into from nitrogen input pipe 4
Flowed out after the tube body 10 in housing 2 from nitrogen efferent duct 4 '.So as to pass through the refrigerant of the circulation in tube body 10
Effectively the biological sample in the sample cell 6 that is placed on the tube body 10 is freezed.Also, inputted by being arranged at nitrogen
The temperature convection that the flow control valve 5 of pipe 4 can be measured effectively based on above-mentioned temperature measuring unit enters the stream of the refrigerant in housing 2
Speed is controlled, to be conducive to keep the temperature the biological sample after freezing.Preferably, as shown in figure 3, above-mentioned tubular body 10 can be with
Be formed as M types, so as to more uniformly freeze.But the shape not limited to this of the tube body 10, as long as can be to loading thereon
Biomaterial is freezed and kept the temperature.And thermal insulation material can be filled in right amount in the gap of the refrigeration pipeline and housing 2, with
Ensure that low temperature is constant.
Above-mentioned refrigeration unit cardinal principle is using the ability of the fast-refrigerating of liquid nitrogen, by sample and the fast quickly cooling of standard solution
Freeze and in required temperature, such as -25 DEG C keep constant state.Therefore for solves the problems, such as refrigeration and insulation can take it is any
Suitable method.
4 to 6 sample cells 6 should be able to be disposably placed in the refrigeration system.Sample cell 6 and refrigeration unit directly connect
Touch, control liquid nitrogen flow velocity to ensure the constant temperature of sample cell 6 in -25 by the flow control valve 5 accessed at nitrogen input pipe 4
℃.Temperature detecting resistance 8 is placed in one end of the sample cell 6, carries out temperature monitoring.I.e., the present invention is with nitrogen as refrigeration source, with
Flow control valve 5 controls nitrogen flow rate, monitors temperature change with temperature detecting resistance 8, keeps it in -25 DEG C of scopes.
With reference to Fig. 2, above-mentioned temperature measuring unit may include the temperature measurer 8 being connected with sample cell 6.In this embodiment, the survey
Warm instrument 8 for example can be the temperature detecting resistance being connected with sample cell 6.
In addition, in the present invention, above-mentioned inductivity coupled plasma mass spectrometry system possesses for the biological sample in housing 2
The icp ms that the gas-carrier pipeline of carrier gas is provided and is connected with the gas-carrier pipeline.The inductive coupling etc. from
Daughter mass spectrograph for example can be icp ms commonly used in the prior art, omit the detailed of its structure herein
Thin description.Carrier gas can be provided to the biological sample in housing 2 by gas-carrier pipeline, so that the biological sample that laser ablation obtains
Aerosol is loaded into ionization in plasma by carrier gas, to carry out subsequent detection.
As shown in Figure 1, above-mentioned gas-carrier pipeline includes the carrier gas inlet portion 3 being respectively arranged on the side wall of housing 2 and carrier gas goes out
Oral area 3 ', carrier gas inlet portion 3 are connected to provide carrier gas, and carrier gas outlet portion into housing 2 with the carrier gas supply source that diagram is omitted
3 ' the icp ms omitted with diagram, which are connected, provides the gas for the biomaterial being loaded into by carrier gas with latter
Colloidal sol, to carry out subsequent detection.
The gas being filled with above-mentioned gas-carrier pipeline includes carrier gas and/or protective gas, and the main function of gas injection is
Completely cut off the disturbing factors such as moisture, air and dust, prevent that sample from volatilizing moisture at low temperatures, and using carrier gas degrading
Particulate bring plasma into.
Present invention also offers a kind of detection method using above-mentioned detection platform, including：
(1) solid biological sample and standard solution will be refrigerated to via refrigeration system, will be carried out by laser ablation system
Laser ablation processing, forms the aerosol of biological sample and standard solution respectively；
(2) by inductivity coupled plasma mass spectrometry system by the aerosol obtained in step (1) be loaded into etc. from
It is detected after being ionized in daughter, the ion-intensity values of the biological sample and standard solution is directly obtained, according to the standard
The ionic strength of solution is worth to the calibration curve on ion-intensity values-concentration of the standard solution, and according to from described
The relational expression that calibration curve obtains converts to obtain the trace member in the biological sample with the ion-intensity values of the biological sample
The content of element.
According to the present invention, under the refrigeration of refrigeration source by biological sample and standard solution rapid condensation into solid-state,
So as to carry out fast and accurately quantitative analysis to sample.Refrigerant system configurations are simple, easy to use, and refrigeration is rapid, and low temperature is protected
Hold constant.And placement multi-disc sample cell 6 can analyze multigroup sample and standard solution within the same time.In unified low temperature
State similarly hereinafter when analyze standard specimen and sample, it is ensured that the stability of data, the accuracy for improving data.The method solves tradition and disappears
The problem of complex for operation step in solution analysis biological sample, and do not introduce any chemical reagents, more environment-friendly high-efficiency, and
Reagent blank when solving the problems, such as sample preparation is excessive, improves detection sensitivity.The method also solves conventional LA-
The problem of being difficult to prepare solid standard specimen in ICP-MS analyses, prepared compared to traditional LA solids standard specimen, solution standard specimen is prepared more
Simply, quickly, accurately.The uniformity of standard specimen has also obtained maximum guarantee.And various loads can be filled with housing 2
Gas, reduces the interference of polyatom particle.
Also, in above-mentioned steps (1), the laser ablation system degrades pattern using line scanning, and the laser is shelled
The wavelength for the laser that erosion system is sent is 213mm, pulse frequency 20Hz, degrades aperture for 15~200 μm, degrades the time for 15
~200 seconds.Since biomaterial belongs to organic-based material, quality is relatively soft and is rich in moisture, therefore selects laser parameter
When need the appropriate energy for reducing laser, increase hot spot.Make that the shadow surface of laser is big, and depth of shine is shallow as much as possible, and
Biomaterial cannot be penetrated.Therefore, by the selection of the parameter of above-mentioned laser ablation system, can effectively to biomaterial into
Row laser ablation.
In addition, in above-mentioned steps (2), the inductivity coupled plasma mass spectrometry system is to jump the time-resolved mode at peak
Gathered data, and in the inductivity coupled plasma mass spectrometry system, RF power is 1000w, sampling depth 9mm, is worked
Gas is argon gas, and plasma flow amount is 15.0L/ minutes, and carrier gas flux is 1.4L/ minutes, data acquisition time 120
Second, the time of integration is 60 seconds., can be effectively to warp by the selection of the parameter of above-mentioned inductivity coupled plasma mass spectrometry system
The aerosol of biomaterial is detected obtained from crossing laser ablation.
Specifically, the present invention can use the trace element in LA-ICP-MS methods measure biological sample, include the following steps：
A) biological sample of suitable size is taken to be placed directly in the groove 7 of sample cell 6；
B) a certain amount of standard solution is drawn, is placed in another sample cell 6, multi-disc sample cell 6 can be increased as needed,
For putting different biological sample or standard solution；And temperature detecting resistance or temperature measurer 8 are connected, multiple surveys can be connected if necessary
Warm resistance or temperature measurer 8；
C) these sample cells 6 are placed on the stainless steel tube 10 of refrigeration unit.Lid 1 is covered and sealed, and is being carried
Helium is filled with air pipe；
D) flow control valve 5 is opened, quickly pours nitrogen, observes the temperature display of temperature measurer 8, when close to -25 DEG C, is subtracted
Few nitrogen pressure, and keep it in -25 DEG C or so；
E) open laser ablation system and pattern is degraded with line scanning, ICP-MS gathers number to jump the time-resolved mode at peak
According to.Calibration curve is made to freeze standard solution, obtains the relational expression of calibration curve.Biological sample is carried out in the same way
Detection is according to the relational expression of foundation and the ionic strength of biological sample, you can containing for trace element in biological specimen is calculated
Amount.
In step e), specifically, the biological sample and the aerosol of standard solution obtained through laser ablation is loaded into
It is detected after being ionized in plasma, the ion-intensity values of biological sample and standard solution is directly obtained, according to standard solution
Ionic strength be worth to the calibration curve on ion-intensity values-concentration of standard solution, and obtained according to from calibration curve
Relational expression convert to obtain the content of the trace element in biological sample with the ion-intensity values of biological sample.
And above-mentioned ion-intensity values for example can separate ion by quadrupole rod, photomultiplier detector detects to obtain, simply
For be exactly with high-velocity electrons to hit gaseous molecular or atom, the cation after ionization is accelerated to import in mass analyzer,
Then it is collected and is recorded by the size order of mass-to-charge ratio (m/z).
The above-mentioned detection method of the present invention can measure the trace element of biological specimen in the case where not clearing up sample, and survey
The result that test result is obtained with micro-wave digestion-inductively coupled plasma mass spectrometry has the uniformity in error range, has
Reasonable.
The present invention is described in more detail by specific embodiment below in conjunction with Fig. 1 to Fig. 3.
Embodiment 1
This gives a kind of laser ablation refrigeration system.Use sus304 stainless steel material systems as shown in Figure 1
Make the box of a upper opening as housing 2, make a call to two apertures in the side surface upper part of housing 2, penetrate stainless steel pipe 3,3 '
And conduit 3,3 ' and 2 firm welding of housing, guarantee are sealed.Wherein, the pressure reducing valve and helium steel that the connection of conduit 3 diagram is omitted
Bottle, to be filled with carrier gas and/or protective gas, and conduit 3 ' is connected with the icp ms that diagram is omitted.
Two holes are made a call in the side bottom of housing 2.By hollow stainless steel tube 10, it is set to be bent to form M shapes by hot-working,
As shown in figure 3, and ensure the smooth no tilting of one side.By this, M types stainless steel tube 10 is smooth is put into upwardly inside housing 2, will not
Both ends, i.e. nitrogen input pipe 4 and the nitrogen efferent duct 4 ' of rust steel pipe 10 are connected with two apertures of the bottom of housing 2 respectively, and
Make itself and 2 firm welding of housing, ensure sealing.And the access flow control valve 5 on the nitrogen input pipe 4.In the stainless steel tube
10 and housing 2 gap in can fill thermal insulation material in right amount.Then, the quartzy glass of a piece of penetrating bubble-free twin polishing can be used
Lid 1 of the glass as refrigeration system, one section of rubber sheet gasket can be installed additional in the junction of lid 1 and housing 2, and makes lid with clip
1 seals with housing 2.
Using the aluminium nitride ceramics of polishing both surfaces as sample cell 6.Processed in the one side of sample cell 6 smooth concave
Groove body 7, for placing sample or standard solution.And set temperature it can also pop one's head in around groove body 7.
This application example tests Cd therein to verify the reliability of apparatus and method using a liver specimens.
It is as follows that method and step micro- in biological sample is measured using LA-ICP-MS methods：
1st, sample cell 6 is positioned in 5% nitric acid and soaks 2h, it is clean with ultrapure water after taking-up and air-dry；
2nd, suitable size, such as the wide liver specimens of long 0.5mm of 1mm or so or so is taken to be placed directly in the groove of sample cell 6
In body 7；
3rd, 2 μ g/g, 4 μ g/g, 6 μ g/g, each 5mL of Cd standard solution of 10 μ g/g are drawn, is placed in four sample cells 6, and
Connect temperature detecting resistance 8；
4th, these sample cells 6 are placed on stainless steel tube 10.Lid 1 is covered and sealed.And in carrier gas input pipe 3
It is filled with helium；
5th, nitrogen flow control valve 5 is opened, nitrogen is quickly filled with the pressure of 0.3mpa, the temperature of observation temperature measurer 8 is shown
Show, when close to -25 DEG C, reduce nitrogen pressure to 0.1mpa, and keep it in -25 DEG C or so.Biological sample and mark at this time
Quasi- solution has frozen；
6th, open laser ablation system and pattern is degraded with line scanning, ICP-MS gathers number to jump the time-resolved mode at peak
According to；
7th, calibration curve is made to freeze standard solution, obtains the relational expression of calibration curve.In the same way to sample
It is detected.According to the relational expression of foundation and the ionic strength of sample, you can micro- in biological specimen contain is calculated
Amount.Wherein, first scanning frost standard solution, obtains one group of element data of a line, it is molten that a standard is obtained after being averaged
The data point of liquid, after testing the standard solution of several concentration respectively, draws calibration curve, then sample is tested, and obtains sample
Corresponding concentration value is calculated by calibration curve after product data.
It is wherein as follows to the optimal setting of each parameter in LA-ICP-MS methods：The ripple of the laser sent in laser ablation system
A length of 213mm, pulse frequency 20Hz, degrade aperture for 200 μm, it is 200s to degrade the time；Inductivity coupled plasma mass spectrometry
In system：RF power is 1000w, and sampling depth 9mm, working gas is argon gas, and plasma flow amount is 15.0L/min,
Carrier gas flux is 1.4L/min, data acquisition time 120s, time of integration 60s.
The data measured in the embodiment 1 are listed in table 1, and the calibration curve in Fig. 4 can be made according to data in table 1.
Table 1
According to table 1 and Fig. 4, standard solution line scan detection 4 points, 4 points be averaged after with average detected intensity
It is worth for ordinate, using the configuration concentration value of Cd as abscissa, can draw corresponding standard curve calibrating curve equation is, calculates
The equation for going out respective alignment curve is y=884.43x, and the related coefficient of calibration curve is 0.996.And sample wire scanning result is such as
Shown in table 2.
Table 2
| First point | Second point | Thirdly | 4th point | 5th point | 6th point | 7th point | |
| Test intensity | 5322 | 7633 | 5479 | 2689 | 5834 | 2699 | 3396 |
| Scaling results | 6.02 | 8.66 | 6.19 | 3.04 | 6.60 | 3.05 | 3.84 |
It can be learnt by as above result, this method standard curve is linearly good, can clearly understand micro in biological sample
The distribution of element.And the method the time it takes is only 1/3rd of traditional analysis, and without any chemistry of introducing
Reagent.The method solves the problems, such as complex for operation step in traditional resolution method analysis biological sample, and does not introduce anyization
Reagent blank when learning reagent, more environment-friendly high-efficiency, and solving the problems, such as sample preparation is excessive, and it is sensitive to improve detection
Degree.The method also solves the problems, such as to be difficult to prepare solid standard specimen in conventional LA-ICP-MS analyses, compared to traditional LA solid marks
Prepared by sample, solution standard specimen prepares more simple, quick, accurate.The uniformity of standard specimen has also obtained maximum guarantee.
Claims (10)
- A kind of 1. detection platform for being used to measure trace element in biological sample, it is characterised in thatThe detection platform includes：The biological sample and standard solution are refrigerated to solid refrigeration system；The laser ablation system for carrying out Direct Laser to being refrigerated to solid biological sample and standard solution and degrading；AndBiological sample and the aerosol of standard solution for laser ablation to be obtained are loaded into ionization in plasma and examine The inductivity coupled plasma mass spectrometry system of the Determination of trace element contents in the biological sample is surveyed,The refrigeration system includes：Store the sample cell of the aluminium nitride ceramics of biological sample and freezing biological sample and standard solution are freezed simultaneously Insulation and the refrigeration unit directly contacted with the sample cell.
- 2. detection platform according to claim 1, it is characterised in that the refrigeration system further includes：There is opening at top and accommodate the housing of at least one sample cell；The lid of the opening and the laser penetration for allowing the laser ablation system to send can be sealed；AndTemperature measuring unit for the temperature for measuring the biological sample.
- 3. detection platform according to claim 2, it is characterised in that the refrigeration unit possesses for into the housing Refrigerant is provided to freeze the refrigeration pipeline of biological sample and the system according to the temperature control that the temperature measuring unit detects The flow control member of the flow velocity of refrigerant in cold pipeline.
- 4. detection platform according to claim 3, it is characterised in that the refrigeration pipeline includes being respectively arranged on the housing Side wall on refrigerant-medium inlet portion and refrigerant outlet portion and be connected to the refrigerant-medium inlet portion and refrigeration be situated between Between matter outlet portion and be arranged in the housing be used for load the tube body of the sample cell；And the flow control member is set In the refrigerant-medium inlet portion.
- 5. detection platform according to claim 2, it is characterised in that the temperature measuring unit includes being connected with the sample cell Temperature measurer.
- 6. detection platform according to claim 2, it is characterised in that the inductivity coupled plasma mass spectrometry system possesses For the inductive coupling for providing the biological sample in the housing gas-carrier pipeline of carrier gas and being connected with the gas-carrier pipeline Plasma mass spectrograph.
- 7. detection platform according to claim 6, it is characterised in that the gas-carrier pipeline includes being respectively arranged on the housing Side wall on carrier gas inlet portion and carrier gas outlet portion, the carrier gas inlet portion is connected with carrier gas supply source, and the carrier gas goes out Oral area is connected with the icp ms.
- A kind of 8. detection method using detection platform according to any one of claim 1 to 7, it is characterised in that bag Include：（1）Solid biological sample and standard solution will be refrigerated to via refrigeration system, laser is carried out by laser ablation system Processing is degraded, forms the aerosol of biological sample and standard solution respectively；（2）By inductivity coupled plasma mass spectrometry system by step（1）The aerosol of middle acquisition is loaded into plasma It is detected after middle ionization, the ion-intensity values of the biological sample and standard solution is directly obtained, according to the standard solution Ionic strength be worth to the calibration curve on ion-intensity values-concentration of the standard solution, and according to from the calibration The relational expression that curve obtains converts to obtain the trace element in the biological sample with the ion-intensity values of the biological sample Content.
- 9. detection method according to claim 8, it is characterised in that in the step（1）In, the laser ablation system Pattern is degraded using line scanning, and the wavelength of laser that sends of the laser ablation system is 213mm, pulse frequency 20Hz, Aperture is degraded for 15~200 μm, degrades the time for 15~200 seconds.
- 10. detection method according to claim 8 or claim 9, it is characterised in that in the step（2）In, the inductive coupling Plasma mass system is to jump the time-resolved mode gathered data at peak, and in the inductivity coupled plasma mass spectrometry system In, RF power is 1000w, and sampling depth 9mm, working gas is argon gas, and plasma flow amount is 15.0L/ minutes, carrier gas Flow is 1.4 L/ minutes, and data acquisition time is 120 seconds, and the time of integration is 60 seconds.
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| CN109668818A (en) * | 2018-12-26 | 2019-04-23 | 宁波大学 | Cell analysis apparatus and method based on laser ablation |
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| CN112362722B (en) * | 2019-07-26 | 2022-07-12 | 中国科学院上海硅酸盐研究所 | Quantitative analysis method for laser ablation inductively coupled plasma mass spectrum |
| CN112213374A (en) * | 2020-08-21 | 2021-01-12 | 苏州格目软件技术有限公司 | Microbial detection platform for aquatic plant growth environment |
| CN112229895A (en) * | 2020-08-21 | 2021-01-15 | 苏州格目软件技术有限公司 | Biological detection platform based on trace element determination |
| CN112229894A (en) * | 2020-08-21 | 2021-01-15 | 苏州格目软件技术有限公司 | Biological trace element detection platform for plasma charge determination |
| CN112229897A (en) * | 2020-09-11 | 2021-01-15 | 苏州格目软件技术有限公司 | Biological detection platform using charge detection |
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| CN102455317A (en) * | 2010-10-27 | 2012-05-16 | 中国石油化工股份有限公司 | Micro component laser ablation isotope analyzing device and method |
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