CN103175727A - Sample preparation instrument - Google Patents
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- CN103175727A CN103175727A CN2013100505815A CN201310050581A CN103175727A CN 103175727 A CN103175727 A CN 103175727A CN 2013100505815 A CN2013100505815 A CN 2013100505815A CN 201310050581 A CN201310050581 A CN 201310050581A CN 103175727 A CN103175727 A CN 103175727A
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Abstract
The invention discloses a sample preparation instrument which comprises an outer shell; a nitrogen blow tank is formed in the top in the shell; the lower part of the nitrogen blow tank is provided with a nitrogen blow plate device; slide ways are respectively arranged at the two sides of the nitrogen blow plate device and in the middle of the nitrogen blow plate device; the nitrogen blow plate device is also provided with nitrogen blow plates capable of sliding along the slide ways; each slide way is provided with a blocking groove, and each nitrogen blow plate is fixed by the corresponding blocking groove; a first fixing groove used for placing a sample liquid receiving plate is formed in the bottom in the outer shell; the first fixing groove is sequentially provided with an intermediate sealing ring and a second fixing groove; the second fixing groove is provided with a sample preparation plate and a solid phase extraction (SPE) bracket; a lever, an oscillator and a negative pressure pump are arranged at the lower part of the first fixing groove; the side surface of the first fixing groove is provided with two small holes which are respectively connected with two fans, and a gas heating device located at the back of the first fixing groove; and the back of the outer shell is provided with a waste liquid outlet hole, an outer nitrogen interface, a negative pressure interface, a power interface and a nitrogen heating device.
Description
Technical field
The present invention relates to chemical analysis and Pharmaceutical Analysis field, relate in particular to a kind of sample preparation instrument.
Background technology
In analytical chemistry and Pharmaceutical Analysis process, sample pre-treatments is the key link of analyzing and testing result precision height, is concentrated determined trace components, improves detection sensitivity and removes the important means that analytic system is had interfering material.The step of sample pretreatment has a great impact credibility, analysis cost and the experimenter's of the high flux of sample detection, data satisfaction.Fast development along with the sample detection technology, for example: LC-MS(Liquid Chromatography-Mass Spectrum, liquid chromatography and mass spectrometry) technology, GC-MS(Gas Chromatography-Mass Spectrum, gas chromatography and mass spectrometry) technology and CE-MS(Capillary Electrophoresis-Mass Spectrum, Capillary Electrophoresis and mass spectrometry) technology, also day by day strengthened for the demand of the fast high-flux of Sample Pretreatment Technique.Therefore, for detection method of exploitation of success, suitable sample-pretreating method of choice and optimization is very important.The trend of at present Sample Pretreatment Technique development be quick, required sample size little, in batches large, automaticity is high, cost is low, labour intensity is low, reagent consumption less, environmental pollution is little and the method pseudo-selectivity is high.Therefore, explore quick, efficient, easy and sample-pretreating method easy robotization has become one of important research direction of current analytical chemistry.
SPE(Solid Phase Extraction, Solid-Phase Extraction) be a kind of novel Sample Pretreatment Technique Used, being combined by liquid-solid extraction and column liquid chromatographic technology develops, the history of existing two more than ten years.SPE compares with traditional liquid-liquid extraction has multinomial superior function, for example: extraction column is disposable throw-away-type, problem without cross pollution, without the emulsification phenomenon, the sample extraction recovery is high, discarded object is easily removed from the laboratory, do not need to use ultrapure solvent, the consumption of organic solvent is few, environmental pollution is little, little to the human injury, required volume of sample is few, processing speed is fast, operate under room temperature, being particularly suited for processing volatility reaches the thermally labile medicine, without being separated operation, easy collection analysis thing component, simple to operate, save time, laborsaving and be easy to the advantage such as robotization, annual growth with 10% enlarges its application always since SPE occurs.
SLE(Supported Liquid-liquid Extraction, the medium liquid-liquid extraction) take traditional liquid-liquid extraction principle as the basis, in conjunction with the form of solid-phase extraction column, utilize that a kind of adsorbability is high, inertia, porous aggregate that specific surface area is large be medium, carries out sample pre-treatments.Detachment process is: water-soluble sample solution is loaded on pillar, (different from Solid-Phase Extraction is, all sample solutions all are retained, n.s. liquid flows out), water-soluble sample liquid is coated on rapidly inertia porous aggregate surface and forms the sample thin film layer, then using with the immiscible organic solvent of water is eluent, target compound is extracted, collect extract and obtain target compound solution.The method has effectively solved the difficult problems such as the complex operation that exists in traditional Liquid-liquid Extraction Processes, time-consuming and easy emulsification, reduced simultaneously the loss of sample, effectively improve the recovery of extraction of sample, and can be combined with 96 orifice plates or 384 orifice plates, realized the high flux sample pre-treatments.
PPT(Protein Precipitation Technology, fast protein precipitation) technology is a kind of principle based on organic solvent deposit protein, combine the form of modern solid-phase extraction column and now membrane technology commonly used develop.Wherein Solid-Phase Extraction is a support of whole technology, and core is film.The film that uses in experiment is a kind of special film, this film does not allow organic solvent or water to pass through at normal temperatures and pressures, in the situation that negative pressure or malleation exist, liquid can pass film and flow out, and oarse-grained material as the denatured protein aggregation, can be retained in the film upside, thereby the protein after realizing precipitating separates with supernatant, reaches biological sample except the purpose of deproteinize.In addition, this technology can be combined with 96 orifice plates or 384 orifice plates, is applied on the robotization solid-phase extracting instrument, thereby realizes high flux and robotization.
Fast filtering technology (Filtration) is a kind of a kind of new fast filtering technology that grows up based on the single filter filtering technique of tradition.The fast filtering technology adopts the form of 96 orifice plates or 384 orifice plates, has realized the high flux pre-treatment of sample, simultaneously, has kept again the form of single solid-phase extraction column, make the selectivity of experimenter in process of the test more, have more dirigibility.Compare with traditional single filter filter type, the fast filtering technology had both kept the advantage of single filter, had overcome again some defectives in single filter filtering technique, such as: complex operation, consuming time, can't realize robotization and high flux etc.So the application of novel fast filtering technology in sample pretreatment process is also more and more extensive.
DPE(Dynamic Packing Extraction, dynamically Solid-Phase Extraction) technology is take solid phase extraction techniques as the basis, prepares in conjunction with quick sample the new technology that theory grows up.Dynamically the core of solid phase extraction techniques is, filler is in bulk in the Tip head or in little solid-phase extraction column, fix with baffle plate up and down, and a space is arranged between baffle plate and filler, this space make filler be bulked in solid-phase extraction column or the Tip head in, and filler is can be within the specific limits movable.Sample pretreatment process is, the Tip head is connected on pipettor or solid phase is extracted post be connected with piston, by repeatedly drawing and getting sample liquid, make testing sample or target compound by filling adsorption, if the compound that is adsorbed on filler is impurity, need not next step processing, if target compound is adsorbed on impurity, need to utilize a kind of suitable eluant, eluent, target compound is eluted, thereby reach the purpose of removing impurity or obtaining target compound.Because filler can move within the specific limits, so in extraction process, the time that sample liquid contacts with filler is compared with common Solid-Phase Extraction process with number of times have been increased a lot, therefore, the efficient that the common Solid-Phase Extraction of speed ratio that dynamic Solid-Phase Extraction reaches extraction equilibrium reaches extraction equilibrium is high a lot, so the method has shortened time for sample pretreatment greatly.Generally speaking, DPE has namely inherited the advantage of traditional Solid-Phase Extraction, has shortened again the time of sample pre-treatments simultaneously, has improved the efficient of sample pre-treatments, has realized new breakthrough on the basis of traditional Solid-Phase Extraction, is with a wide range of applications.
The method of SPE, SLE, PPT, Filtration and five kinds of sample pretreatments of DPE is that method the most frequently used in sample pretreatment is also state-of-the art sample pretreating method.
The inventor finds to exist at least in prior art following shortcoming and defect in realizing process of the present invention:
At present, in most of assay laboratory, these five kinds of Sample Pretreatment Technique Useds have application, and major part is all the medical grade polypropylene injection tube form of single tube, need manual operation.And for manual operation, have the following disadvantages: personnel interfere the too many poor repeatability of easily introducing error, sample, can't realize that the organic reagent that uses in robotization and high flux and sample preparation process is also larger to the injury of human body.At present, 96 orifice plates or 384 orifice plate Sample Pretreatment Technique Useds, frequency of utilization is also fewer in the assay laboratory of China or scientific research institution, and main cause is there is no supporting high flux sample preparation instrument mutually; Even high-throughout sample preparation instrument is arranged, a plurality of instruments such as needed Nitrogen evaporator and shaker in subsequent step so, not only expensive but also shared spatial volume is larger, is unfavorable for actual application.
Summary of the invention
The invention provides a kind of sample preparation instrument, the present invention realized semi-automatic so that automatically high flux carry out the experiment of various sample pre-treatments, avoided the introducing of error, improved the recovery of sample, guaranteed the repeatability of sample preparation, simultaneously, also kept and implemented traditional single tube Solid-Phase Extraction and liquid-liquid extraction isotype, seen for details hereinafter and describe:
A kind of sample preparation instrument comprises: shell, it is characterized in that, and described shell inner top is provided with nitrogen and blows groove, is provided with nitrogen in the bottom that described nitrogen blows groove and blows panel assembly; Both sides and centre position that described nitrogen blows panel assembly also are respectively arranged with slideway, blow at described nitrogen the nitrogen that arranges on panel assembly along described slideway slip and blow plate, are provided with draw-in groove on described slideway, and the fixing described nitrogen of described draw-in groove blows plate; Described shell inner bottom part is provided with for the first pickup groove of placing the sample liquid dash receiver, is disposed with intermediate seal circle and the second pickup groove on described the first pickup groove, is provided with sample preparation plate and SPE support on described the second pickup groove; On described the first pickup groove sidewall, three holes are arranged, wherein two holes are connected with the electric fan that is placed in the first pickup groove back, gas-heating apparatus is positioned at the electric fan bottom, airtight cavity of the common composition of another hole and electric fan and Solid-Phase Extraction the first pickup groove, the second pickup groove, intermediate seal circle and solid phase extraction sample plate provides the circulating-heating air; Described the first pickup groove base apertures is connected by inner conduit with the negative pressure pump that is positioned at the instrument rear lower; Be provided with waste liquor stream outlet, nitrogen external tapping, negative pressure interface, power interface and gas-heating apparatus at the described shell back side.
Described nitrogen blows groove and comprises: variable valve,
Described nitrogen blows panel assembly and is arranged on described nitrogen by described variable valve and blows on groove.
The model that described nitrogen blows plate is specially: 96 holes, 24 holes, 12 holes or 9 holes; Described nitrogen blows the model of plate and the model of described Solid-Phase Extraction column holder is complementary.
Described nitrogen gas heating arrangement is comprised of the heating cylinder body of two series connection.
Also be provided with the nitrogen flow table in described shell, be connected with extraneous source nitrogen by described nitrogen external tapping by nitrogen conduit;
Also be provided with negative pressure dynamics indicator gauge on described shell, show vacuum magnitude.
The back side of described shell also is provided with fan.
The bottom of described the first pickup groove also is provided with aperture, is connected with negative pressure pump by described aperture to realize the bottom negative pressure.
The side of described the first pickup groove also is provided with aperture, is connected with electric fan, gas-heating apparatus by described aperture to realize bottom malleation recirculated hot air.
Described electric fan is connected with the first pickup groove by the aperture of the side of the first pickup groove.
The side aperture of described the first pickup groove, electric fan, gas-heating apparatus, the first pickup groove, the second pickup groove, intermediate seal circle, solid phase extraction sample plate, airtight cavity of common formation provides bottom malleation recirculated hot air thereby blow over journey for nitrogen.
Also be provided with oscillation device below described the first pickup groove and described the second pickup groove, described oscillation device comprises: lever and oscillator,
Described the first pickup groove be connected the second pickup groove and connect described oscillator by described lever.
When the mode of operation of described sample preparation instrument was the programmed control operation, described sample preparation instrument also comprised: display control panel,
By the regulation and control of described display control panel realization to liquid feeding, positive/negative-pressure conversion, time-delay, nitrogen blowing temp.
Described nitrogen blows the rifle head assembly that plate is specially 96 holes.
The model of described Solid-Phase Extraction column holder is specially: 1mL, 3mL, 6mL or 10mL;
When the model of described Solid-Phase Extraction column holder was 1mL, 1mL Solid-Phase Extraction column jecket was arranged by 3 * 8 and is placed; When the model of described Solid-Phase Extraction column holder was 3mL, 3mL Solid-Phase Extraction column jecket was arranged by 4 * 6 and is placed; When the model of described Solid-Phase Extraction column holder was 6mL, 6mL Solid-Phase Extraction column jecket was arranged by 3 * 4 and is placed; When the model of described Solid-Phase Extraction column holder was 10mL, 10mL Solid-Phase Extraction column jecket was arranged by 3 * 3 and is placed.
The beneficial effect of technical scheme provided by the invention is: the present invention can be simultaneously with high flux nitrogen blowing apparatus, rocking equipment with move subsidiary function in the sample preparations such as liquid and the core technology of sample preparation has organically been joined together, be integrated on same instrument; In addition, this instrument can also be easily with two yuan of above Sample Pretreatment Technique Useds or polynary combining, thereby realize the pre-service to complex sample; The exchange function of single tube and 96-orifice plate makes sample preparation instrument have adaptability widely; The design of its Subminature multifunctional is easy to carry and movement, has saved simultaneously valuable lab space; Avoid the introducing error, improved the repeatability of sample; Adding sample preparation instrument can also directly be connected with computing machine, makes it more convenient to use effectively, so this multifunctional all high flux sample preparation instrument is a brand-new multi-functional sample preparation new instrument of cover, is with a wide range of applications.
Description of drawings
Fig. 1 is the Facad structure schematic diagram of a kind of sample preparation instrument provided by the invention;
Fig. 2 is the structure schematic diagram of a kind of sample preparation instrument provided by the invention;
Fig. 3 is the side structure schematic diagram of a kind of sample preparation instrument provided by the invention;
Fig. 4 is a kind of sample preparation instrument provided by the invention bottom solid-phase extraction device part right section structural representation;
Fig. 5 is vertical view on 1mL Solid-Phase Extraction column holder provided by the invention;
Fig. 6 is vertical view on 1mL solid-phase extraction column frame bottom provided by the invention hole;
Fig. 7 is vertical view on 3mL Solid-Phase Extraction column holder provided by the invention;
Fig. 8 is vertical view on 3mL solid-phase extraction column frame bottom provided by the invention hole;
Fig. 9 is vertical view on 6mL Solid-Phase Extraction column holder provided by the invention;
Figure 10 is vertical view on 6mL solid-phase extraction column frame bottom provided by the invention hole;
Figure 11 is vertical view on 10mL Solid-Phase Extraction column holder provided by the invention;
Figure 12 is vertical view on 10mL solid-phase extraction column frame bottom provided by the invention hole.
In accompanying drawing, the list of parts of each label representative is as follows:
1: shell; 2: variable valve;
3: nitrogen blows groove; 4: nitrogen blows panel assembly;
5: nitrogen blows plate; 6: the second pickup grooves;
7: the first pickup grooves; 8: the intermediate seal circle;
9: sample preparation plate and SPE support; 10: display control panel;
11: slideway; 12: draw-in groove;
13: the nitrogen flow table; 14: negative pressure dynamics indicator gauge;
15: gas-heating apparatus; 16: fan;
17: the waste liquor stream outlet; 18: the nitrogen external tapping;
19: the negative pressure interface; 20: power interface;
21: oscillator; 22: lever;
23: negative pressure pump; 24: electric fan;
25: gas-heating apparatus; 26: the side vent port;
27: the electric fan hole;
A:1mL column jecket hole; Bottom, B:1mL column jecket hole;
C:3mL column jecket hole; Bottom, D:3mL column jecket hole;
E:6mL column jecket hole; Bottom, F:6mL column jecket hole;
F:10mL column jecket hole; Bottom, G:10mL column jecket hole.
Embodiment
For making the purpose, technical solutions and advantages of the present invention clearer, embodiment of the present invention is described further in detail below in conjunction with accompanying drawing.
Therefore, for the problem that exists in present sample pretreatment, the problem that has solved for the sample preparation field of succeeding in developing of high flux and automatic multifunctional sample preparation instrument.The embodiment of the present invention just for these problems that exist in the sample pretreatment process, has been developed a kind of multifunctional all high flux sample preparation instrument.
For realize semi-automatic so that automatically high flux carry out various sample preparations experiment, avoid introducing error, improve the repeatability of sample, the embodiment of the present invention provides a kind of sample preparation instrument, referring to Fig. 1 to Fig. 4, sees for details hereinafter and describes:
A kind of sample preparation instrument comprises: shell 1, shell 1 inner top are provided with nitrogen and blow groove 3, are provided with nitrogen in the bottom that nitrogen blows groove 3 and blow panel assembly 4; Both sides and centre position that nitrogen blows panel assembly 4 also are respectively arranged with slideway 11, blow at nitrogen the nitrogen that arranges on panel assembly 4 along slideway 11 slips and blow plate 5, are provided with draw-in groove 12 on slideway 11, and draw-in groove 12 fixed nitrogens blow plate 5; Shell 1 inner bottom part is provided with for being disposed with on intermediate seal circle 8 and the second pickup groove 6, the second pickup grooves 6 on the first pickup groove 7, the first pickup grooves 7 of placing the sample liquid dash receiver and is provided with sample preparation plate and SPE support 9; On the first pickup groove 7 sides, electric fan hole 27 and vent port 26 are arranged; There is electric fan 24 27 back, electric fan hole; Gas-heating apparatus 25 is arranged at electric fan 24 bottoms; The first pickup groove 7, the second pickup groove 6, intermediate seal circle 8, sample preparation plate and SPE support 9 and electric fan 24, gas-heating apparatus 25 and concussion fill 21 and put airtight cavity of common composition; Lever 22 and rocking equipment 21 are arranged at the first pickup groove 7 bottoms; Be provided with waste liquor stream outlet 17, nitrogen external tapping 18, negative pressure interface 19, power interface 20 and nitrogen gas heating arrangement 15 at housing 1 back side.
Wherein, during specific implementation, realize that by nitrogen external tapping 18 nitrogen that nitrogen blows groove 3 blows; Export 17 derivation that realize waste liquid by waste liquor stream; Realize the energising of sample preparation instrument by power interface 20.Waste liquor stream outlet 17 and negative pressure interface 19 are connected to form circuit system by surge flask; In experimentation, the waste liquid that produces flows out by the aperture that is positioned at the first pickup groove 7 bottoms.
Wherein, during specific implementation, can also regulate by slideway 11 and draw-in groove 12 height that nitrogen blows plate 5.
Wherein, nitrogen blows groove 3 and comprises: variable valve 2, nitrogen blow panel assembly 4 and are arranged on nitrogen by variable valve 2 and blow on groove 3.During specific implementation, variable valve 2 can be the stationary installations such as bolt.
Further, in order to satisfy the multiple needs in practical application, the model that nitrogen blows plate 5 is specially: 96 holes, 24 holes, 12 holes or 9 holes.Should guarantee in practical application that nitrogen blows the model of plate 5 and the model of Solid-Phase Extraction column holder is complementary.
When the model of blowing plate 5 when nitrogen is 96 hole, be 8 * 12; When the model of blowing plate 5 when nitrogen is 24 hole, be 3 * 8 or 4 * 6; When the model of blowing plate 5 when nitrogen is 12 hole, be 3 * 4; When the model of blowing plate 5 when nitrogen is 9 hole, be 3 * 3.Blow plate 5 for the nitrogen of placing above-mentioned different model on the second pickup groove 6.
Wherein, top nitrogen blowing heating arrangement 15 is comprised of the heating cylinder body 15 of two series connection.
Further, for the nitrogen flow that nitrogen external tapping 18 is poured is counted, also be provided with nitrogen flow table 13 in shell 1, be connected with extraneous source nitrogen by nitrogen external tapping 18 by nitrogen conduit, show concrete airshed size by the nitrogen flow table 13 that is positioned on shell 1.
Further, for the pressure of show sample preparing instrument inside, also be provided with negative pressure dynamics indicator gauge 14 on shell 1, show vacuum magnitude.
Form negative pressure by the negative pressure pump 23 that is placed in instrument internal, be used for showing that the vacuum meter of vacuum magnitude is by being placed in negative pressure dynamics indicator gauge 14 realizations on the instrument front panel.
Further, for the airshed that makes sample preparation instrument inside is disperseed more even, sample can dry up as soon as possible, also be provided with aperture 27 in the side of the first pickup groove 7 and be connected with back electric fan 24, utilize side exhaust hole 26, electric fan 24 bottom heating devices 25, the first pickup groove 7, the second pickup groove 6, intermediate seal circle 8 and sample preparation plate and airtight cavity of the common formation of SPE support 9 that the function of circulating-heating gas is provided.
Further, for the heat radiation of supplementary instrument, also be provided with fan 16 at the back side of shell 1.
Further, for the recovery that improves sample and the reappearance of method, also be provided with oscillation device below the first pickup groove 7 and the second pickup groove 6, oscillation device comprises: lever 22 and oscillator 21, the first pickup grooves 7 and the second pickup groove 6 are connected with oscillator 21 by lever.During specific implementation, the frequency by vibration and time are realized the control to oscillator 21.
Wherein, the mode of operation of this sample preparation instrument: manually operation and programmed control move two kinds of patterns.Manual mode of operation, in the whole process of sample pretreatment, can be by the button at PC interface, change at any time the temperature of heating, the time-delay of operation, the amount of 96 hole rifle head imbititions, stream of nitrogen gas amount, vacuum tightness size, vacuum time-delay, the temperature of auxiliary heating gas and time-delay.Program control mode, the action in above each step is controlled by PC, and instrument can move according to the step of having set voluntarily, until complete whole program.
When the mode of operation of sample preparation instrument was the programmed control operation, this sample preparation instrument also comprised: display control panel 10, and by the regulation and control of display control panel 10 realizations to liquid feeding, positive/negative-pressure conversion, time-delay, nitrogen blowing temp.The process that realizes is, by the whole program of instrument internal is carried out software programming, integrated on a chip, external presentation is a LCDs, realizes control to all programs of whole device by display screen.
The sample preparation instrument that the embodiment of the present invention provides can be regulated airshed by external gas source, and by instrument itself with the airshed size used of can be the at any time online demonstration experiment of nitrogen flow table 13, thereby can regulate and control as soon as possible; By instrument with nitrogen blow panel assembly 4, can realize that different size nitrogen blows the replacing of plate 5, nitrogen blows the O-ring seal on panel assembly 4, the nitrogen that can guarantee different holes blows plate 5, such as: 96 holes (8 * 12), 24 holes (3 * 8), 24 holes (4 * 6), 12 holes (3 * 4), 9 holes (3 * 3) etc. different size, fit like a glove with sample preparation instrument.By the nitrogen gas heating arrangement 15 that sample preparation instrument inside carries, can realize needed nitrogen temperature in experiment is controlled; By slideway 11 and draw-in groove 12, realize that nitrogen blows the fixing of plate 5, and the height and position that nitrogen blows plate 5 is adjusted.
Referring to Fig. 5 to Figure 12, the embodiment of the present invention also designs the SPE support in sample preparation plate and SPE support 9, and the model of Solid-Phase Extraction column holder is specially: the Solid-Phase Extraction column holder of 1mL, 3mL, 6mL or 10mL.
When the model of Solid-Phase Extraction column holder was 1mL, 1mL Solid-Phase Extraction column jecket was arranged by 3 * 8 and is placed; When the model of Solid-Phase Extraction column holder was 3mL, 3mL Solid-Phase Extraction column jecket was arranged by 4 * 6 and is placed; When the model of Solid-Phase Extraction column holder was 6mL, 6mL Solid-Phase Extraction column jecket was arranged by 3 * 4 and is placed; When the model of Solid-Phase Extraction column holder was 10mL, 10mL Solid-Phase Extraction column jecket was arranged by 3 * 3 and is placed.
The Solid-Phase Extraction column holder in 96 holes adopts general support in this area, and the embodiment of the present invention is not done at this and given unnecessary details.
Wherein, during specific implementation, the O-ring seal on the O-ring seal around the first pickup groove 7, the first pickup groove 7, the second pickup groove 6, the second pickup groove 6, and intermediate seal circle 8 has formed solid-phase extraction device.By changing sample pretreatment solid-phase extraction plate of different nature and realizing the pre-service of different samples from the dash receiver that the sample preparation plate is complementary, for example the design of the SPE support of 96 holes (8 * 12), 24 holes (3 * 8), 24 holes (4 * 6), 12 holes (3 * 4), 9 holes (3 * 3) has realized the conversion of the solid-phase extraction column placement plate of specification, thereby has adapted to the demand of different sample preparations.The existence of intermediate seal circle 8 has realized the sealing of the first pickup groove 7 and the second pickup groove 6 on the solid-phase extraction device, has guaranteed the sealing in the experimentation, has solved the problem of vacuum magnitude.By the first middle aperture in pickup groove 7 bottoms, the waste liquid that produces in the sample pretreatment process can be derived by inner conduit and the waste liquor stream that is connected with the external waste collection bottle outlet 17 that is positioned at the back side, and with waste collection, reduce the pollution to environment and people.By the circuit system between the aperture on the first pickup groove 7 sides, inner conduit, waste liquor stream outlet 17, negative pressure pump 23 and negative pressure interface 19, realize the negative pressure that the sample pretreatment process is required, show the size of negative pressure by negative pressure dynamics indicator gauge 14 simultaneously, thereby can regulate accordingly.
The principle of work of this sample preparation instrument is specially: diversified nitrogen blows the design of plate 5, sample after pre-service can being completed directly dries up, and does not need to shift, and has reduced the chance of sample contamination, saved simultaneously the space, and by the dual regulation to airshed and temperature, realize for different material, the nitrogen of different dynamics blows, enlarged the range of application of instrument, in addition, can adopt two kinds of patterns of programmed control and manual operation to carry out nitrogen and blow control, reduce the man power and material.
During specific implementation, nitrogen blows plate 5 can be the rifle head assembly in 96 holes, adds internal components to the control of rifle head assembly accuracy, has realized the effect that the 96 passage volley of rifle fires move liquid and application of sample, has greatly saved time and labour.By regulating different gears, the inhalant liquid scale of construction is controlled.By the rifle head automatic spring assembly in the design of the lower end of rifle head assembly, reduced automatic sample and the time of removing in rifle head process in the sample pretreatment process, improved sample pretreatment speed, increased efficient.
The robotization solid-phase extraction device has been realized all processes of sample semi-automation and even robotization sample pretreatment.Comprise the steps such as the pre-service of extraction column, loading, drip washing and wash-out.In addition, the design of the Solid-Phase Extraction of semi-automation and even robotization part, the technology of repeatedly switching of employing negative pressure and malleation can be carried out pre-service or drip washing to extraction column at any time, has increased the Modulatory character in the process; The introducing of nitrogen heating arrangement 15 has realized the control to nitrogen blowing temperature, has greatly shortened sample and has dried up the time of using, and can be combined with the malleation of dash receiver part simultaneously, and both direction carries out drying up of liquid, has improved the pre-treating speed of sample.Simultaneously, this sample preparation instrument has also kept implements traditional single tube Solid-Phase Extraction, support liquid-liquid extraction isotype, can also realize the semi-automatic or full-automatic process of single tube Solid-Phase Extraction.The exchange function of this single tube and 96-orifice plate makes this sample preparation instrument have adaptability widely.
The concussion function, concussion depends on solid-phase extraction device and realizes, and concussion can guarantee the homogeneity that sample mixes in preprocessing process, improve the recovery of sample and the reappearance of method.
Embodiment 1:SPE operating process
C18 bonded silica gel filler, specification are the operating process of 96-well plate of the 2mL in 100mg filler/hole:
The placement of sample preparation 96 hole C18 filler plates: the 96 hole C18 filler plates of 2mL are fixed on the second pickup groove 6 of the upper pre-processed board placement of solid-phase extraction device, make it and fit like a glove on every side.Receiving trap below noting is not put for the time being, guarantees that 96 orifice plates and surrounding fit like a glove, to guarantee the sealing of the whole part of Solid-Phase Extraction.
Activation: every hole adds 2mL methyl alcohol, after adding 1mL for the first time, after balance 1min, vacuumizes and makes its outflow, then add the methyl alcohol of 1mL to activate, and vacuumizes after balance 1min and makes its outflow.Then every hole adds the water of 2mL, and the step that adds according to methyl alcohol activates.After activation is completed, vacuumize activating solution is flowed out fully, complete the reactivation process of pillar.
Loading: pipette concentration and be in the hole of benzoic acid aqueous solution 1mL loading to 96 orifice plate of 500 μ g/mL, sample solution drips under nature under Action of Gravity Field, if a lower speed is very slow, can adopt the acceleration loading process of assisted vacuum, and the vacuum dynamics is controlled to be-0.5bar.The principle of loading is, the speed that sample liquid is lower is can not ether fast, guarantee that the absorption of sample on pillar can reach balance, and is also can not ether slow, otherwise can extend analysis time, and general 1mL sample liquid needs 1-3min.
Drip washing: draw with 96 row's liquid transfer gun heads of upper part the sample preparation plate that the water of 1mL joins in sample preparation plate and SPE support 9 and carry out drip washing; repeat 2 times; if the lower speed of leacheate is very slow; start the carrying out of the auxiliary lessivation of negative pressure function; leacheate is flowed out, and the vacuum dynamics is controlled to be-0.5bar.
With taking off of sample preparation plate and SPE support 9, dash receiver is placed in the trough of solid-phase extraction device the first pickup groove 7, sample preparation plate and SPE support 9 are restored, be placed in original position.Attention: guarantee sealing.
Wash-out: the methyl alcohol that pipettes 1mL with 96 rifle heads joins in sample preparation plate in sample preparation plate and SPE support 9, make naturally lower of leacheate, if a lower speed is very slow, vacuumize auxiliary drip washing, leacheate is flowed in dash receiver, repeatedly repeat this process several times, finally need the methyl alcohol of 3mL that the complete drip washing of sample is got off.
Dry up: sample preparation plate and SPE support 9 are removed, following dash receiver is taken out, be placed in the position of sample preparation plate and SPE support 9, nitrogen dries up.Simultaneously, the bottom can adopt heated air to assist to dry up.The stream of nitrogen gas amount is 0.6mL/min.The temperature of lower heating is controlled to be 30 ℃.
Redissolve: the distilled water or the damping fluid that pipette 1mL join in sample well, ultrasonic redissolution.
Embodiment 2: medium liquid-liquid extraction (SLE) operating process
The operating process of medium liquid-liquid extraction, the specification of 96 hole SLE plates used is: amount of filler is the SLE96 orifice plate of the 2mL in 200mg/ hole.
Place dash receiver: dash receiver is placed in the trough that solid-phase extraction device is placed the first pickup groove 7 of dash receiver.
The placement of sample preparation plate 96 hole SLE plates: the 96 hole SLE plates of 2mL are fixed on the second pickup groove 6 of the upper pre-processed board placement of solid-phase extraction device, make it and fit like a glove on every side.
Loading: the concentration that pipettes 200 μ L is the propylparaben aqueous solution of 200 μ g/mL, is loaded in SLE post hole, and note: maximum applied sample amount should guarantee that sample can not flow out, balance 5min.
Extraction: the methylene chloride that pipettes 1mL with 96 rifle heads joins in sample preparation plate and SPE support 9, makes naturally lower of leacheate, and leacheate flows in dash receiver, repeatedly repeats this process several times, finally needs the methylene chloride of 3-5mL that the complete drip washing of sample is got off.If a lower speed is very slow, vacuumize auxiliary drip washing, the vacuum dynamics is controlled to be-0.5bar, continues 10-15s.
Dry up: (being determined on a case-by-case basis) removes 96-well plate sample preparation plate and SPE support 9, and following dash receiver is placed on the second pickup groove 6 of placing sample preparation plate and SPE support 9, and nitrogen dries up, under add that hot-air is auxiliary to be dried up.Nitrogen air blowing flow is 0.6mL/min, and the temperature of lower heating is controlled to be 30 ℃.
Redissolve: if sample need to dry up processing, distilled water or the damping fluid that need pipette 1mL join in sample well, ultrasonic redissolution.
Embodiment 3: novel protein sedimentation operating process
Fast protein sedimentation operating process, the specification of 96 orifice plates used are the penetrating shape plate of 1mL.
Put into dash receiver: dash receiver is placed on the first pickup groove 7 that solid-phase extraction device is placed dash receiver.
The placement of sample preparation plate 96 hole PPT plates: the second pickup groove 6 places with the 96 hole PPT plates of 1mL are fixed on that the upper pre-processed board of solid-phase extraction device places guarantee that 96 orifice plates and surrounding fit like a glove, with the sealing of the whole part of assurance Solid-Phase Extraction.
Loading: the acetonitrile that pipettes 1mL with 96 rifle heads joins in 96 hole PPT sample preparation plates, the more pending sample solution that pipettes 200 μ L joins in the sample well that contains the 1mL acetonitrile, and concussion 2min precipitates protein fully.
Vacuumize: vacuumize liquid is flowed out fully.The vacuum tightness size is controlled to be-0.8bar.Attention: the dynamics that vacuumizes can not be too large, otherwise film is leaked, and causes sample to become muddy.
Dry up (being determined on a case-by-case basis): remove the 96 hole PPT sample preparation plates that are positioned at sample preparation plate and SPE support 9 places, following dash receiver is placed in the position of sample preparation plate, nitrogen dries up, under add that hot-air is auxiliary to be dried up.Nitrogen air blowing flow is 0.6mL/min, and the temperature of lower heating is controlled to be 30 ℃.
Redissolve: the distilled water or the damping fluid that pipette 1mL join in sample well, ultrasonic redissolution.
Embodiment 4: the operating process of dynamic Solid-Phase Extraction (DPE)
Dynamically solid phase extracts operating process: the Tip specification: the SCX amount of filler is the rifle head of 50mg/1mL
Activation: the DPE post is connected on the liquid-transfering gun of 1mL, sucks 0.5mL methyl alcohol and appropriate air, make the abundant mixing of filler and solution, after mixing, methyl alcohol is got, then sucked the water of 0.5mL, mixing, activation.
Loading: place the little reagent bottle of a 2mL below the DPE post, contain 0.6 sample solution in reagent bottle, suck sample, and air is passed through, from but sample fully mix with filler, balance 25 seconds is got, carry out again loading, 2 times repeatedly, until sample all is sucked out.
Drip washing: suck successively each 1mL washing of methanol-water DPE pillar, at last aqueous solution is got.
Wash-out: the ammonification methanol solution 1mL of absorption 5% is in dash receiver, repeatedly suck and get 3 times, to guarantee that melamine is eluted fully, at last the eluent in dash receiver is dried up, dash receiver is placed on the second pickup groove 6, dry up, regulating the upper part nitrogen flow is 0.6mL/min, below temperature be controlled to be 30 ℃, bottom hot gas flow be instrument itself with airshed, need not regulate.
Redissolve: add the methanol aqueous solution of the 50:50 of 1mL, ultrasonic redissolution is filtered, and is to be detected.
In sum, the embodiment of the present invention provides a kind of sample preparation instrument, the embodiment of the present invention can be simultaneously with high flux nitrogen blowing apparatus, rocking equipment with move subsidiary function in the sample preparations such as liquid and the core technology of sample preparation has organically been joined together, be integrated on same instrument; In addition, this instrument can also be easily with two yuan of above Sample Pretreatment Technique Useds or polynary combining, thereby realize the pre-service to complex sample; The exchange function of single tube and 96-orifice plate makes sample preparation instrument have adaptability widely; The design of its Subminature multifunctional is easy to carry and movement, has saved simultaneously valuable lab space; Avoid the introducing error, improved the repeatability of sample; Adding sample preparation instrument can also directly be connected with computing machine, makes it more convenient to use effectively, so this multifunctional all high flux sample preparation instrument is a brand-new multi-functional sample preparation new instrument of cover, is with a wide range of applications.
It will be appreciated by those skilled in the art that accompanying drawing is the schematic diagram of a preferred embodiment, the invention described above embodiment sequence number does not represent the quality of embodiment just to description.
The above is only preferred embodiment of the present invention, and is in order to limit the present invention, within the spirit and principles in the present invention not all, any modification of doing, is equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. sample preparation instrument comprises: shell (1), it is characterized in that, and described shell (1) inner top is provided with nitrogen and blows groove (3), is provided with nitrogen in the bottom that described nitrogen blows groove (3) and blows panel assembly (4); Both sides and centre position that described nitrogen blows panel assembly (4) also are respectively arranged with slideway (11), blow at described nitrogen the nitrogen that arranges on panel assembly (4) along described slideway (11) slip and blow plate (5), be provided with draw-in groove (12) on described slideway (11), the fixing described nitrogen of described draw-in groove (12) blows plate (5); Described shell (1) inner bottom part is provided with for the first pickup groove (7) of placing the sample liquid dash receiver, be disposed with intermediate seal circle (8) and the second pickup groove (6) on described the first pickup groove (7), be provided with sample preparation plate and Solid-Phase Extraction column holder (9) on described the second pickup groove (6); Be provided with waste liquor stream outlet (17), nitrogen external tapping (18), negative pressure interface (19), power interface (20) and nitrogen heating arrangement (15) at described shell (1) back side, there is two fans of device (24) and the gas-heating apparatus (25) that recyclegas is provided at described the first pickup groove (7) back; Described two fans (24) are that the first pickup groove (7) provides hydronic gas by two apertures (27) and the exhaust hole (26) that is positioned on the first pickup groove (7) sidewall; Be provided with oscillator (21), lever (22) and negative pressure pump (23) bottom described the first pickup groove (7).
2. a kind of sample preparation instrument according to claim 1, is characterized in that, described nitrogen blows groove (3) and comprising: variable valve (2), described nitrogen blow panel assembly (4) and are arranged on described nitrogen by described variable valve (2) and blow on groove (3).
3. a kind of sample preparation instrument according to claim 1, is characterized in that, the model that described nitrogen blows plate (5) is specially: 96 holes, 24 holes, 12 holes or 9 holes; Described nitrogen blows the model of plate (5) and the model of described Solid-Phase Extraction column holder is complementary.
4. a kind of sample preparation instrument according to claim 1, is characterized in that, described nitrogen heating arrangement (15) is comprised of the heating cylinder body of two series connection.
5. a kind of sample preparation instrument according to claim 1, is characterized in that, also is provided with nitrogen flow table (13) in described shell (1), is connected with extraneous source nitrogen by described nitrogen external tapping (18) by nitrogen conduit; Also be provided with negative pressure dynamics indicator gauge (14) on described shell (1), show vacuum magnitude; The back side of described shell (1) also is provided with fan (16).
6. a kind of sample preparation instrument according to claim 1, is characterized in that, the bottom of described the first pickup groove (7) also is provided with aperture, realizes the bottom negative pressure by described aperture and negative pressure pump (23); The side of described the first pickup groove (7) also is provided with three apertures, wherein aperture (27) and electric fan (24) directly communicate, aperture (26), aperture (27), electric fan (24) and solid-phase extraction device be common forms an airtight box, thereby dries up the heated air that process provides circulation for Solid-Phase Extraction.
7. a kind of sample preparation instrument according to claim 1, it is characterized in that, also be provided with oscillation device below described the first pickup groove (7) and described the second pickup groove (6), described oscillation device comprises: lever (22) and oscillator (21), described the first pickup groove (7) be connected the second pickup groove (6) and connect described oscillator (21) by described lever (22).
8. a kind of sample preparation instrument according to claim 1, is characterized in that, when the mode of operation of described sample preparation instrument was the programmed control operation, described sample preparation instrument also comprised: display control panel (10),
By the regulation and control of described display control panel (10) realization to liquid feeding, positive/negative-pressure conversion, time-delay, nitrogen blowing temp.
9. according to claim 1 or 3 described a kind of sample preparation instruments, is characterized in that, described nitrogen blows the rifle head assembly that plate (5) is specially 96 holes.
10. a kind of sample preparation instrument according to claim 1, is characterized in that, the model of described Solid-Phase Extraction column holder is specially: 1mL, 3mL, 6mL or 10mL;
When the model of described Solid-Phase Extraction column holder was 1mL, 1ml Solid-Phase Extraction column jecket was arranged by 3 * 8 and is placed; When the model of described Solid-Phase Extraction column holder was 3mL, 3mL Solid-Phase Extraction column jecket was arranged by 4 * 6 and is placed; When the model of described Solid-Phase Extraction column holder was 6mL, 6mL Solid-Phase Extraction column jecket was arranged by 3 * 4 and is placed; When the model of described Solid-Phase Extraction column holder was 10mL, 10mL Solid-Phase Extraction column jecket was arranged by 3 * 3 and is placed.
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| CN103743607A (en) * | 2014-01-20 | 2014-04-23 | 厦门大学 | Chip card box for pretreatment before food sample detection |
| CN104148126A (en) * | 2014-08-20 | 2014-11-19 | 南京中科神光科技有限公司 | Self-adaption sucking head withdrawing device |
| CN104436753A (en) * | 2014-12-09 | 2015-03-25 | 天津博纳艾杰尔科技有限公司 | Solid phase extractor |
| CN106442092A (en) * | 2016-11-03 | 2017-02-22 | 睿科仪器（厦门）有限公司 | Drawer device capable of automatically inserting nitrogen-blowing needle into concentration test tube |
| CN108680426A (en) * | 2018-05-16 | 2018-10-19 | 天津博纳艾杰尔科技有限公司 | A kind of nitrogen evaporator |
| CN114200064A (en) * | 2020-09-18 | 2022-03-18 | 江苏豪思睦可生物科技有限公司 | Sample pretreatment system |
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