CN111889155A - Multi-channel electrospray micro-fluidic chip and application thereof - Google Patents
Multi-channel electrospray micro-fluidic chip and application thereof Download PDFInfo
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Abstract
The invention discloses a multi-channel electrospray micro-fluidic chip and application thereof, wherein the multi-channel electrospray micro-fluidic chip comprises two first gas phase channels and at least two liquid phase channels, and the liquid phase channels are arranged between the two first gas phase channels; the nozzle of the first gas phase flow channel and the nozzle of the liquid phase flow channel are arranged at intervals. In the process of electrospray, the relative positions of the liquid-phase nozzle and the first gas-phase nozzle are fixed, so that the spraying conditions are stable and consistent, on one hand, the extraction function of macroscopic electrospray can be realized, the extraction efficiency and the extraction effect of the extraction liquid on the liquid to be detected are improved, and the quick detection on a complex sample is favorably realized; on the other hand, the problem that double liquid paths or multiple liquid paths are difficult to simultaneously carry out electrospray is solved, more application forms are finally created for the combination of the micro-fluidic chip and the mass spectrometer, and the method can be widely applied to the aspects of substance rapid mixing reaction, protein folding and stretching analysis, internal standard test and the like.
Description
Technical Field
The invention relates to the field of micro-fluidic chips for electrospray, in particular to a multi-channel electrospray micro-fluidic chip and application thereof.
Background
For mass spectrometry, single-path sample ionization has difficulty in meeting the requirements of relevant tests, and a two-path or even multi-path ionization mode has become a research hotspot. The dual-path or multi-path spraying ion source can be applied in various aspects, such as accurate mass number determination through dual-path internal standards, extraction and detection of complex samples through dual-path spraying, signal intensity enhancement of multi-path spraying, sensitivity improvement and the like, which cannot be realized through single-path spraying.
In the application research of double-path spraying, an extraction electrospray ion source (EESI) plays an important role in mass spectrometry. Because when complex samples such as urine, serum, lake water and milk are directly sprayed into a mass spectrometer by using an electrospray ion source for signal detection, adduct formation, sample residue and non-volatile matter accumulation can be rapidly caused, and finally, the sensitivity of the mass spectrometer is rapidly reduced and difficult to recover. EESI can not need any sample pretreatment process, realizes the quick on-line detection to complicated sample such as material such as urine, milk.
But when adopting EESI to carry out the double-circuit extraction spraying, the stability and the sensitivity of test can be influenced to the locating position and the contained angle of two nozzles, lead to the extraction inefficiency of the liquid that awaits measuring usually, and the ionization effect is poor to lead to that the test signal is weak, experiment operability and reproducibility are relatively poor.
Accordingly, the prior art is yet to be improved and developed.
Disclosure of Invention
In view of the defects of the prior art, the present invention aims to provide a multi-channel electrospray microfluidic chip and an application thereof, which aims to improve the extraction efficiency and effect of mass spectrometry test liquid, thereby enhancing the test signal of a mass spectrometer to the liquid to be tested.
The technical scheme of the invention is as follows:
a multi-channel electrospray microfluidic chip, comprising: two first gas phase flow channels and at least two liquid phase flow channels; the liquid phase flow channels are arranged between the two first gas phase flow channels; the nozzle of the first gas phase flow channel and the nozzle of the liquid phase flow channel are arranged at intervals.
The multi-channel electrospray microfluidic chip further comprises at least one second gas-phase flow channel positioned between the at least two liquid-phase flow channels, and the nozzles of the second gas-phase flow channel and the nozzles of the liquid-phase flow channels are arranged at intervals.
The multi-channel electrospray micro-fluidic chip is characterized in that the other end of each liquid phase flow channel is provided with a liquid phase inlet.
The multi-channel electrospray micro-fluidic chip is characterized in that the other ends of the two first gas-phase flow channels and the second gas-phase flow channel are converged to form a gas-coherent flow channel, and a gas-phase inlet is formed at the tail end of the gas-coherent flow channel.
The multi-channel electrospray micro-fluidic chip is characterized in that the nozzles of two adjacent liquid phase flow channels are arranged at intervals.
The multi-channel electrospray micro-fluidic chip is characterized in that the interval between the nozzle of the first gas-phase flow channel and the nozzle of the liquid-phase flow channel and the interval between the nozzle of the second gas-phase flow channel and the nozzle of the liquid-phase flow channel are both 10-100 micrometers.
The multi-channel electrospray micro-fluidic chip is characterized in that the central axes of the nozzles of the liquid phase flow channel, the first gas phase flow channel and the second gas phase flow channel are parallel, and the nozzles are on the same plane.
The multi-channel electrospray microfluidic chip is characterized in that the depths of the first gas-phase flow channel and the second gas-phase flow channel are greater than that of the liquid-phase flow channel.
The multichannel electrospray micro-fluidic chip is made of PDMS or PMMA.
Use of a multi-channel electrospray microfluidic chip as described above for extractive electrospray of a liquid or for multi-channel simultaneous electrospray.
Has the advantages that: the invention provides a multi-channel electrospray micro-fluidic chip which comprises two first gas phase channels and at least two liquid phase channels, wherein the liquid phase channels are arranged between the two first gas phase channels; the nozzle of the first gas phase flow channel and the nozzle of the liquid phase flow channel are arranged at intervals. In the process of electrospray, the relative positions of the liquid-phase nozzle and the first gas-phase nozzle are fixed, so that the spraying conditions are stable and consistent, on one hand, the extraction function of macroscopic electrospray can be realized, the extraction efficiency and the extraction effect of the extraction liquid on the liquid to be detected are improved, and the quick detection on a complex sample is favorably realized; on the other hand, the problem that double liquid paths or multiple liquid paths are difficult to simultaneously carry out electrospray is solved, more application forms are finally created for the combination of the micro-fluidic chip and the mass spectrometer, and the method can be widely applied to the aspects of substance rapid mixing reaction, protein folding and stretching analysis, internal standard test and the like.
Drawings
Fig. 1 is a schematic structural diagram of a multi-channel electrospray microfluidic chip according to a first embodiment of the invention.
Fig. 2 is a schematic structural diagram of a multi-channel electrospray microfluidic chip according to a second embodiment of the invention.
Fig. 3 is a schematic structural diagram of a multi-channel electrospray microfluidic chip according to a third embodiment of the invention.
Fig. 4 is a schematic structural diagram of a multi-channel electrospray microfluidic chip according to a fourth embodiment of the invention.
Fig. 5 is a schematic diagram of a hybrid taylor cone spray formation.
Fig. 6 is a schematic diagram of a split taylor cone spray formation.
FIG. 7a is a mass spectrum of rhodamine B in urine solution tested using an EESI ion source.
FIG. 7B is a mass spectrum of a urine solution of rhodamine B tested using the first embodiment of the invention.
Fig. 8 is a graph of total ion flux stability for a spray using a third embodiment of the invention.
FIG. 9a is a mass spectrum of a single-channel electrospray rhodamine B solution.
Figure 9b is a mass spectrum of reserpine solution from single-channel electrospray.
FIG. 9c is a mass spectrum of a double-channel simultaneous electrospray rhodamine B and reserpine solution in accordance with a third embodiment of the present invention.
Detailed Description
The invention provides a multi-channel electrospray micro-fluidic chip and application thereof, and the invention is further described in detail below in order to make the purpose, technical scheme and effect of the invention clearer and more clear. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In the process of mass spectrometry, the extraction and ionization effects of the liquid to be tested have great influence on the test effect of mass spectrometry, but because the relative positions of two independent nozzles of a macroscopic electrospray extraction ionization (EESI) device are not fixed, the relative positions of the two independent nozzles need to be adjusted before the experiment, the operability and reproducibility of the experiment are often reduced, the extraction efficiency of liquid electrospray extraction by adopting the EESI device is not high, mass spectrometry test signals are easily low, and the test result is influenced.
Referring to fig. 1 and fig. 2, the multi-channel electrospray microfluidic chip provided by the present invention includes: two first gas phase flow channels 10 and at least two liquid phase flow channels 20; the liquid phase flow channels 20 are both arranged between the two first vapor phase flow channels 10; the nozzle of the first gas phase flow passage 10 and the nozzle of the liquid phase flow passage 20 are arranged at an interval
The invention can perform extraction electrospray ionization on complex liquid to be detected, wherein the complex liquid to be detected refers to complex liquid with various compositions which is not extracted yet, such as urine containing an object to be detected. Referring to fig. 5, in the using process of the microfluidic chip, the liquid phase channels 20 are used for guiding the liquid, wherein at least one of the liquid phase channels 20 is used for guiding the liquid to be detected, and the other liquid phase channels 20 are used for guiding the extraction liquid, and only high voltage needs to be applied to the extraction liquid. Because the liquid phase flow channel 20 has a small interval, in the process of extracting electrospray, the liquid to be detected and the extraction liquid can be mixed at the nozzle of the liquid phase flow channel 20, so that the extraction and ionization of the extraction liquid on the liquid to be detected are realized, a mixed Taylor cone spray is formed, and the atomization is carried out simultaneously. Because the micro-fluidic chip has high integration level, the relative position of the liquid flow channel to be detected and the extraction liquid flow channel is fixed, and the nozzle interval is small, the spraying conditions of the two liquids are stable and consistent, and the atomization effect is good. The liquid phase flow passage 20 positioned in the middle is better used for guiding the liquid to be tested, so that a Taylor cone spraying mode for mixing the extraction liquid and the liquid to be tested can be formed at the spray opening of the liquid phase flow passage 20, the extraction efficiency and the extraction effect of the extraction liquid on the liquid to be tested are further improved while the two liquids are uniformly mixed, and the subsequent test signal is enhanced. The first gas phase flow channel 10 is used for guiding gas, the first gas phase flow channel 10 is distributed on two sides of the liquid phase flow channel 20, so that the mixed liquid can be prevented from spreading to two sides at the nozzle of the liquid phase flow channel 20 to influence the spraying effect, meanwhile, the mixed liquid can be further atomized, and the ionization efficiency is improved. The arrangement of the first interval can obtain a mixed Taylor cone with a proper size, so that the spraying effect of the micro-fluidic chip is further improved. The method effectively realizes the detection of the complex liquid to be detected, reduces the pretreatment process of the liquid to be detected, improves the extraction efficiency of the complex liquid to be detected, and enhances the ionization capacity of the liquid to be detected.
In one embodiment, the nozzles of two adjacent liquid phase flow channels 20 are spaced, preferably, the width of the nozzle spacing of the liquid phase flow channels 20 is 20 μm, and the spacing is small, so as to facilitate the mixing of the liquid to be detected and the extraction liquid.
In another embodiment, at least one second gas phase flow channel 30 is further included and is located between the at least two liquid phase flow channels 20, that is, the second gas phase flow channel 30 is used for separating the liquid phase flow channels 20, and the nozzle of the second gas phase flow channel 30 is spaced from the nozzle of the liquid phase flow channel 20, so that a structure in which a plurality of independent nozzles are arranged side by side can be formed, wherein an independent nozzle refers to a nozzle formed by arranging at least one liquid phase flow channel 20 between two gas phase flow channels (including the first gas phase flow channel 10 and/or the second gas phase flow channel 30). For example, two first gas phase flow channels 10, three liquid phase flow channels 20, and one second gas phase flow channel 30 may be used, wherein the second gas phase flow channel 30 is disposed between any pair of adjacent liquid phase flow channels 20, so that two independent nozzles are formed, wherein one independent nozzle has two nozzles of the liquid phase flow channels 20, and the other independent nozzle has only one nozzle of the liquid phase flow channels 20; it is also possible to arrange the structure as shown in fig. 4, using two first vapor flow paths 10, three liquid phase flow paths 20, and two second vapor flow paths 30, wherein two second vapor flow paths 30 are arranged between adjacent liquid phase flow paths 20, forming three independent nozzles each having a nozzle of only one liquid phase flow path 20.
In a preferred embodiment, referring to fig. 3 and 4, the second gas phase flow channels 30 are spaced from the adjacent liquid phase flow channels 20 to form a structure in which the second gas phase flow channels 30 are alternately spaced from the liquid phase flow channels 20, that is, only one nozzle of the liquid phase flow 20 is formed in each individual nozzle. Thus, during extractive electrospray, multiple individual nozzles form multiple smaller separate taylor cone sprays, as shown in fig. 6. In the process of extracting and electrically spraying the complex liquid to be detected, voltage is only applied to the extraction liquid, and the liquid to be detected and the extraction liquid form separated Taylor cone spray at the nozzles of the liquid phase flow passages 20 where the liquid to be detected and the extraction liquid are respectively positioned, and the atomization is respectively carried out. Under the atomization state, the extraction liquid drops and the liquid drops to be detected collide with each other, so that the extraction and ionization of the extraction liquid on the liquid to be detected are realized. It should be noted that, the flow channel selection of the extraction liquid and the liquid to be tested can be performed according to the needs and the spraying effect. The device can also be used for double-liquid-path or multi-liquid-path electrospray ionization, generally speaking, the double-path or multi-path electrospray ionization can reduce the ion suppression effect, so that the mass number can be accurately determined through the internal standard, the mass spectrum signal intensity can be enhanced, the sensitivity is improved, and the high-efficiency and high-throughput analysis of a macromolecular sample is realized. However, when two nozzles are simultaneously electrospray, it is often difficult to simultaneously electrospray two or more nozzles due to the strong electric field interference of the nozzle tips with each other. As the gas phase flow channels are arranged between the adjacent liquid phase flow channels 20, the liquid phase flow channels 20 are independent from each other, and the shape of the tip of a common nozzle is almost not existed, so that the mutual electric field interference is small, the simultaneous electrospray of two-way or multi-way nozzles can be realized, and the spray stability is high. It should be noted that, when performing two-liquid-path or multi-liquid-path electrospray, since the liquid to be detected is not extracted in the spraying process, the liquid to be detected needs to be extracted or purified in advance, and then electrospray is performed. When the two-liquid-path or multi-liquid-path electrospray is performed, the liquid-phase flow channels 20 are used for guiding the liquid to be detected, and high voltage is applied to the liquid to be detected for performing electrospray, wherein the liquid to be detected can be the same or different, and mass spectrograms of different objects to be detected can be obtained simultaneously by performing electrospray on different liquids to be detected simultaneously, so that the detection efficiency is improved.
The multi-channel electrospray micro-fluidic chip further expands the coupling range of the micro-fluidic chip and a mass spectrometer, can be widely applied to the aspects of substance rapid mixing reaction, protein folding and stretching analysis, internal standard test and the like, is really favorable for realizing rapid detection of complex samples, gets rid of the complicated sample pretreatment process in a laboratory, and makes a certain contribution to the development of mass spectrometer miniaturization and portability.
Referring to fig. 1 to 4, the other end of each liquid phase flow channel 20 is provided with a liquid phase inlet 21, and the liquid phase inlets 21 of different liquid phase flow channels 20 are different, so that liquid enters the liquid phase flow channel 20 from the liquid phase inlet 21, and taylor cone spray is formed at the nozzle of the liquid phase flow channel 20. The other ends of the two first gas phase channels 10 and the other end of the second gas phase channel 30 converge to form a gas coherent channel 40, a gas inlet 41 is disposed at the end of the gas coherent channel 40, that is, the two first gas phase channels 10 and all the second gas phase channels 30 share one gas inlet 41, and the gas enters the gas coherent channel 40 from the gas inlet 41 and separately passes through each of the first gas phase channels 10 and the second gas phase channels 30 at the disposed position.
In one embodiment, the nozzle interval between the nozzle of the first gas phase flow channel 10 and the nozzle of the liquid phase flow channel 20 and the nozzle interval between the nozzle of the second gas phase flow channel 30 and the nozzle of the liquid phase flow channel 20 are both 10 to 100 μm, preferably 15 to 25 μm. A certain interval is required between the outlet of the liquid phase flow passage 20 and the nozzle of the gas phase flow passage (including the first gas phase flow passage 10 or the second gas phase flow passage 30) to form taylor cone spray with a proper size, and the taylor cone spray formed due to the large interval is large and unstable in spray, so that the spray effect is influenced; the small interval easily causes the phenomenon that a gas phase flow channel and a liquid phase flow channel are mixed due to the fact that the upper half piece and the lower half piece are not firmly bonded in the preparation process of the chip.
In one embodiment, the central axes of the orifices of the liquid-phase flow passage 20, the first gas-phase flow passage 10, and the second gas-phase flow passage 30 are parallel, and the orifices are all on the same plane. That is, the sections of all the flow channels close to the nozzles are distributed in parallel, and the gas phase and the liquid phase are simultaneously sprayed out. The parallel distribution can make the first gas phase runner 10 and the second gas phase runner 30 on every liquid phase runner 20 or all liquid phase runner 20 both sides completely symmetric distribution, can make liquid spout simultaneously in parallel completely like this, when just facing the mass spectrometer entry with the liquid phase spout, the parallel spun liquid phase can as much as possible and more accurately get into the mass spectrometer, guarantees that the signal can not reduce because of the inaccurate detection signal that leads to of spraying direction.
In one embodiment, the first vapor phase channel 10 and the second vapor phase channel 30 have a depth greater than the depth of the liquid phase channel 20. The first vapor phase channel 10 and the second vapor phase channel 30 are deeper, which facilitates atomization of the liquid and prevents spreading of the liquid at the nozzle of the liquid phase channel 20.
In one embodiment, the multichannel electrospray microfluidic chip is made of PDMS or PMMA. The invention adopts a multilayer overlay soft lithography method to process a microfluidic chip, and Polydimethylsiloxane (PDMS) or polymethyl methacrylate (PMMA) is used as a processing material. The soft photoetching process flow of the multi-channel electrospray micro-fluidic chip mainly comprises the working procedures of whirl coating, exposure, development, mold entering, curing, mold stripping, cutting, punching, bonding and the like, and the process is mature, and is not described in detail in the invention. Specifically, an upper half piece and a lower half piece of the microfluidic chip can be formed respectively, and the upper half piece and the lower half piece are both provided with a micro-channel structure, so that the upper half piece and the lower half piece are combined and processed into a complete chip in a mode of bonding PDMS and PDMS. The processing technology can be changed according to different application requirements of the microfluidic chip. The method has the advantages of simple processing technology, easy bonding, low cost, convenient large-scale preparation and the like.
The invention also provides application of the multi-channel electrospray extraction micro-fluidic chip, and the multi-channel electrospray extraction micro-fluidic chip is mainly used for extraction electrospray of liquid or simultaneous electrospray of multiple liquid paths to generate micro atomized liquid drops for a mass spectrometer to detect.
As shown in figure 7a and figure7b, it can be known from the figure that the rhodamine signal intensity and the signal-to-noise ratio obtained by adopting the EESI ion source are far lower than those of the ion source test of the microfluidic chip. Total ion flux (mass to charge ratio 443-. As shown in fig. 8, it can be seen that the total ion current intensity of the two samples was only 1.6% relative to the standard deviation (RSD) during the two-pass electrospray process, which fully demonstrates the stability of the two-pass simultaneous electrospray. As shown in FIGS. 9a, 9b and 9c, it can be seen that the mean signal intensities of rhodamine and reserpine were 1.1X 10, respectively, when two single-pass sprays were applied separately5And 4.8X 105. However, the conditions are kept unchanged, two samples are added on the two paths of the microfluidic chip at the same time, high voltage is applied, and when simultaneous independent spraying is carried out, the signal intensity of rhodamine is hardly influenced, and the signal intensity of reserpine is slightly reduced. This shows that during simultaneous spraying, two-way simultaneous electrospray has certain influence, including electric field intensity interference and ion suppression effect etc. but total influence is less, can not influence mass spectrum signal test basically.
In summary, the present invention provides a multi-channel electrospray microfluidic chip, which includes two first vapor phase channels and at least two liquid phase channels, wherein the liquid phase channels are disposed between the two first vapor phase channels; the nozzle of the first gas phase flow channel and the nozzle of the liquid phase flow channel are arranged at intervals. In the process of electrospray, the relative positions of the liquid-phase nozzle and the first gas-phase nozzle are fixed, so that the spraying conditions are stable and consistent, on one hand, the extraction function of macroscopic electrospray can be realized, the extraction efficiency and the extraction effect of the extraction liquid on the liquid to be detected are improved, and the quick detection on a complex sample is favorably realized; on the other hand, the problem that double liquid paths or multiple liquid paths are difficult to simultaneously carry out electrospray is solved, more application forms are finally created for the combination of the micro-fluidic chip and the mass spectrometer, and the method can be widely applied to the aspects of substance rapid mixing reaction, protein folding and stretching analysis, internal standard test and the like.
It is to be understood that the invention is not limited to the examples described above, but that modifications and variations may be effected thereto by those of ordinary skill in the art in light of the foregoing description, and that all such modifications and variations are intended to be within the scope of the invention as defined by the appended claims.
Claims (10)
1. A multi-channel electrospray microfluidic chip, comprising: two first gas phase flow channels and at least two liquid phase flow channels; the liquid phase flow channels are arranged between the two first gas phase flow channels; the nozzle of the first gas phase flow channel and the nozzle of the liquid phase flow channel are arranged at intervals.
2. The multi-channel electrospray microfluidic chip according to claim 1, further comprising at least one second gas-phase flow channel positioned between the at least two liquid-phase flow channels, wherein the orifices of the second gas-phase flow channel are spaced apart from the orifices of the liquid-phase flow channels.
3. The multi-channel electrospray microfluidic chip according to claim 2, wherein the other end of each liquid phase flow channel is provided with a liquid phase inlet.
4. The multi-channel electrospray microfluidic chip according to claim 2, wherein the two first gas-phase flow channels and the other end of the second gas-phase flow channel converge to form a gas-coherent flow channel, and a gas-phase inlet is arranged at the tail end of the gas-coherent flow channel.
5. The multi-channel electrospray microfluidic chip according to claim 1, wherein the nozzles of two adjacent liquid phase flow channels are spaced apart.
6. The multi-channel electrospray microfluidic chip according to claim 2, wherein the nozzle interval of the first gas-phase flow channel and the nozzle interval of the liquid-phase flow channel and the nozzle interval of the second gas-phase flow channel and the nozzle interval of the liquid-phase flow channel are both 10-100 μm.
7. The multi-channel electrospray microfluidic chip according to claim 2, wherein the central axes of the orifices of the liquid phase flow channel, the first gas phase flow channel and the second gas phase flow channel are parallel and the orifices are all on the same plane.
8. The multi-channel electrospray microfluidic chip according to claim 2, wherein the first and second gas phase flow channels have a depth greater than the liquid phase flow channel.
9. The multi-channel electrospray microfluidic chip according to any of claims 1-8, wherein the multi-channel electrospray microfluidic chip is made of PDMS or PMMA.
10. Use of a multi-channel electrospray microfluidic chip according to claim 9 for extractive electrospray of a liquid or for multi-channel simultaneous electrospray.
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