GB2604084A - Multi-channel sampling and detection device - Google Patents
Multi-channel sampling and detection device Download PDFInfo
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Abstract
A multi-channel sampling and detection device, comprises; two multi-channel sampling devices (A+B) and one gas detection device; each multi-channel sampling device includes a multi-channel gas path automatic switching unit, a sample detection channel, an interval gas supply device connected to the switching unit or said sample detection channel; wherein each multi-channel sampling device includes multiple expiratory chamber units of mice and multiple front buffer gas chambers each installed at gas output ends of the expiratory chambers to pre-collect samples of mice exhaled breath. The interval gas is used to purge the detection channel.
Description
Multi-channel Sampling and Detection Device
Technical field
The present invention provides a multi-channel gas sampling and detection device with interval gas supply. Specifically, the interval gas can delay the diffusion of gas samples and to prevent cross-interference between them as much as possible, the interval gas is used to achieve continuous detection of multiple gas samples within a short time range. It is especially suitable for connecting with real-time online detecting equipment. This device can be widely used in the detection of various gases, such as environmental atmosphere, human exhaled breath, animal exhaled breath, cell headspace, etc.
Background technology
As a potential research method, gas analysis has received extensive attention from the scientific community and society. It has certain applications in the fields of environmental atmospheric monitoring, early screening, and pharmacokinetics. Exhaled breath contains thousands of volatile and non-volatile substances that can directly reflect the current state of the human body, tissues, cells, microorganisms, etc., because most of them come from metabolism, environmental exposure or therapeutic intervention. In addition, the breath analysis has good patient compliance because the exhaled breath can be quickly, conveniently, and continuously unlimited non-invasive sampling.
At present, breath analysis has aroused extensive scientific attention and great interest of clinicians, and has shown great potential in disease diagnosis, early screening, drug detection, and drug abuse analysis. For example, Sinues et al. conducted studies on the effects of morning and evening administration on the metabolism of ketamine in mice (Gauging circadian variation in ketamine metabolism by real-time breath analysis, Chemical Communications. 2017; Drug phannacokinetics determined by real-time analysis of mouse's exhalation. Angewandte Chemie international Edition, 2015). Based on high-resolution mass spectromety (quadrupole time-of-flight mass spectrometry) and 2 Limin dry air as the carrier gas, a single mouse exhaled breath can be detected online in real time within 1 hour to obtain high time resolution (monitoring time resolution <1 s) exhaled breath metabolism data. However, the sample throughput and detection efficiency of this method are low, and it cannot meet the needs of multiple sample collection and detection in environmental atmospheric monitoring, environmental exposure, early screening, and phannacokinetics.
Zhu J et al, used rapid detection and analysis of mouse exhaled breath to identify acute bacterial lung infections. (Secondary electrospray ionization-mass spectrometry (SES1-MS) breath printing of multiple bacterial lung pathogens, a mouse model study, Journal of Applied Physiology, 2013). In the experiment, there were six parallel samples for the control group mice and the normal group mice, taking into account the individual differences and reproducibility of the objects. The mice were anesthetized in sequence and placed on a ventilator to collect their exhaled breath in a Tedlar bag. The gas sample was detected by SESE-MS in the positive ion mode once with in 1 hour, and the average spectrum was taken for data analysis. This analysis method will cause different storage time (sampling bag adsorption) or reaction time (adsorbent adsorption) between parallel samples, which will have a certain impact on the data results.
The current multi-channel gas sample collection device has solved the problem of simultaneous collection of multiple samples. Patent JP 200267098 A provides a multi-channel sampling device for animal exhaled breath. The device includes an animal storage container, an exhalation collection port, an exhalation collection channel, an exhalation transfer device, an exhalation storage container, and a gas detection device. The following is the process of a single-channel animal exhaled breath collection device: animal exhaled breath is collected from the exhalation collection port through the exhalation collection channel to the exhalation transfer device, and then transferred to the exhalation storage container for storage and waiting to be detected. The device integrates multiple single-channel animal exhalation collection devices into one to achieve simultaneous collection of multiple animal exhalation samples.
Patent CN 202204709U provides a fully automatic multi-channel gas path switching atmospheric sampling device, which including an atmospheric sampler, a rotary multi-channel gas path switching device, and a control module. The atmospheric sampler is connected to the gas port of the rotary multichannel gas path switching device through a silicone tube, and the other end is connected to the gas collection device. The sample collection channel can be selectively opened to collect the gas sample into the designated sampling bag. The rotary multi-channel gas path switching device has multiple gas paths, and the sampling time, sampling channel, and sampling time can be set according to work requirements. The control module determines the channel position of the rotary multi-channel gas path switching device according to these parameters and the signal input by the sensor. Then by controlling the start and stop of the motor to control the connection or closing of each channel during the sampling process of the atmospheric sampler, the collection of multiple samples can be completed within a certain period of time.
Simultaneous collection of multiple gas samples can be achieved by the above-mentioned two multi-channel sampling devices, but for the subsequent detection of multiple gas samples, the current main method is to sequentially send the collected multiple gas samples to the gas detection device for detection. Moreover, a cleaning operation is generally required between the two gas samples, which is time-consuming and cannot meet the requirement of continuous detection of multiple gas samples. After the detection of the previous gas sample is completed, the sampling path and the detection gas chamber will be cleaned in order to avoid the sample residue in the sampling path and the detection gas chamber from interfering with the detection result of the subsequent gas sample. The cleaning gas is charged into the sampling path and the detection gas chamber for a period of cleaning, and then discharged from the exhaust port or pumped out of the cavity with a pump. After the cleaning is completed, the next gas sample detection is performed. in the patent CN 104111228 A, it is mentioned that after the gas sample detection is completed. the cleaning gas needs to be filled into the sampling path and the detection gas chamber by an air pump until the gas chamber is filled and then cleaned for 200 seconds.
Another method for detecting multiple gas samples is that some gas detection devices have multiple detection channels. However, considering the cost, instrument structure, detection method and other factors, most gas detection device currently has only one sample detection channel, such as high-resolution quadrupole time-of-flight mass spectrometry, gas chromatography mass spectrometry (GC-MS), etc. Therefore, the detection method of multiple gas samples is mainly to sequentially detect multiple gas samples collected.
Term specification
1. Interval gas: delaying the diffusion of each gas sample and preventing cross-interference between the two samples as much as possible. Gases such as carbon dioxide (CO2), oxygen (02), nitrogen (N2), and inert gases are common components of interval gas.
2. Cross interference: The mixing between the two samples changed the composition of the original sample and caused some interference to the subsequent sample detection and analysis.
3. Gas sample: is a shapeless, volumetric, compressible and swellable fluid that is actually observed or investigated in research.
4. Sample throughput: refers to the number of samples that can be detected per unit time.
5. Detection efficiency: is the ratio of detection output to detection time 6. Diffusion: The process in which molecules of a certain substance enter other substances through irregular movement and diffusion movement.
7. Animal exhaled breath: refers to the breath exhaled by animals placed in the self-made breathing chamber.
8. Environmental atmosphere. refers to a thick layer of ahnospherie molecules gathered around the earth, called the atmosphere.
9. Cell headspace: refers to the gas produced by cell metabolism during cell culture.
Cleaning gas: The gas used to clean the gas channel or the residual substance in the gas chamber. The main function is to prevent the residual from interfering with the detection result of the next sample.
1 I Dry clean air: refers to the mixed gas in the atmosphere except for water vapor, liquid and solid particles, referred to as dry air 12 Zero gas: refers to gases that do not contain components to be detected or interfering substances, but are allowed to contain components that are not related to the detection.
13 Carrier gas: Refers to the Teflon (RTM) tube used in the breathing chamber to connect with the gas that meets the requirements of the experiment. On the one hand, it provides the animal with normal exhalation, arid on the other hand, it discharges excess exhaust gas. The carrier gas must not contain target compounds and substances that will interfere with the detection results.
14 Concentrated gas sample: A high-pressure or high-speed gas sample whose concentration is increased by physical means (such as compression, pressurization, etc.).
Concentrated interval gas: which refers to high-pressure interval gas after increasing the air pressure by physical means (such as compression, pressurization, etc.) 16 Multi-channel: refers to the parallel use of multiple sampling devices, or the sequential injection of multiple sampling channel, which can achieve high-throughput detection of gas samples, effectively shortening the detection time arid detection cycle.
17 Public channel: refers to the gas channel shared by multiple gas samples.
18 Monitoring time resolution: refers to the period of repeated monitoring of animal exhalation.
19 Compliance: refers to the patient's behavior in accordance with the doctor's prescription and advice. It is customary to call the patient "cooperation"; othenvise, it is called non-compliance.
Teflon (RTM) (polytetrafluoroethylene, PTFE): generally called "non-stick coating" or "easy to clean material". This material is resistant to acids, alkalis, and various organic solvents, and is almost insoluble in all solvents. At the same time, PTFE has the characteristics of high temperature resistance, and its friction coefficient is very low, so it can be used for lubrication and become an ideal coating for easy-to-clean pipelines.
21 Pharmacokinetics: It is a subject that quantitatively studies the discipline of absorption, distribution, metabolism and excretion of drugs in organisms, and uses mathematical knowledge to explain the laws of blood drug concentration changes over time.
Summary of invention
A basic multi-channel sampling device is shown in Figure I, the device includes a gas inlet diversion unit (10), an expiratory chamber unit of mice (20), and a multi-channel gas path automatic switching unit (30), Motor control module. The gas inlet diversion unit is connected to the mouse expiratory chamber unit through a Teflon (RTM) tube, and the other end is connected to a carrier gas supply device that meets the experimental requirements for the mouse to breathe. The multi-channel gas path automatic switching unit is equipped with rotary table, an electronic control module and a high-precision mass flow controller which can accurately control die gas collection flow rate.
The multi-channel sampling control program software sets parameters such as carrier gas flow rate, sampling sequence, sampling time and flow rate to meet sampling requirements. The electric control module guides the rotation of the rotary table to switch the gas path and connects the target sample collection path to the sample detection channel (400) of the gas detection device (40) for real-time online analysis. During the collection of the target mouse exhaled breath sample, the other mouse exhaled breath channels are in an empty state (the carrier gas and mouse exhaled breath are discharged from the exhaust port), and the carrier gas will be continuously provided to the mouse to breathe to maintain life. Generally, Plexiglass (RTM) tube or centrifuge tube (PTFE) is used as the expiratory chamber unit of mice, shown in Fig. 2.
The carrier gas enters through the gas inlet for the mouse to breathe, and the mouse exhaled breath is loaded into the storage device for storage or gas detection equipment for detection. At present, animal exhaled breath and human breath analysis experiments have been extensively carried out in the research fields of pharmacokinetics and early screening. Since this type of experiment is mainly analyzed by the quality and quantity changes of volatile metabolites, it is the best choice to observe the changes in exhaled breath in real time. Since exhaled breath sample has low concentration and is difficult to store, it is generally required that exhaled breath sample can be collected and detected in real time.
The multi-channel sampling device can realize the collection of multiple gas samples. Special design is needed to achieve this purpose. Fig.3 and Fig.4 are examples of disc-shaped equidistant gas inlet diversion chamber and rotary multi-channel gas path conversion mechanism.
Whether it is single-channel gas sample collection or multi-channel gas sample collection, the general process is shown in Fig.5: start to collect gas samples according to sampling requirements after cleaning all sampling pipelines. There are two detection methods to choose after a gas sample is collected. One is real-time online analysis. The sample collection channel is directly connected to the detection channel of die gas detection device, and the gas sample is directly detected after collection. The other is offline analysis, where the collected gas samples are collected in a storage device and stored under certain conditions, and then the gas samples are detected. It is mainly aimed at pre-processing or storing gas samples that have little interference with subsequent gas sample detection results, and the premise of the method is that the gas samples have suitable storage methods and devices.
An improved multiple gas sample detection device is shown in Figure 6: multiple gas samples flow in turn into the gas detection device; to avoid interference of sample residue on the next sample detection, channel cleaning is needed. However, the detection of samples is intermittent and time-consuming because of long sampling channel or detection chamber cleaning. The cleaning gas does not participate in the detection and is discharged directly from the exhaust gas port and the degree of cleanliness can only be judged by experience. The current condition and contamination of the sample channel can hardly be observed in real time so the experimental adjustment cannot be made to ensure the efficiency of the experiment.
Some gas samples need to be detected within a short time range, such as some atmospheric samples, unstable and prone to reaction or light easily decomposition; and some biological exhaled breath sample concentration is low, in particular gas diffusion during the testing process resulting in the loss of certain targets below the instrument's detection limit, and some gas samples that arc difficult to store or have stricter storage conditions. Because of the loss rate in the sample storage process, it may also result in the loss of certain objects in the sample, or the background of the gas sample storage device will cause greater interference to the sample, such as the need to achieve real-time on-line continuous detection of gas samples in a short period of time (most biological exhalation samples belong to this category). How should a gas detection device with only one sample channel achieve the need to continuously analyze multiple gas samples over a period of time? and how do multiple samples meet the diffusion of continuous sample delay gas samples and preventing cross-interference between them as much as possible when converged through a public channel.
In order to solve these problems, the present invention discloses a multi-channel sampling and detection device, when multiple samples need to converged through a public channel or only one sample detection channel of the device needs to continuously analyze multiple samples at the same time, to delay the diffusion of gas samples and preventing cross-interference between them as much as possible whether for real-time on-line detection or offline sampling analysis of multiple gas samples.
A multi-channel gas sampling and detection device is shown in Figure 6. it includes a gas inlet diversion unit, a multi-channel gas path automatic switching unit (30), a control unit or control system (not shown in figure) and an interval gas supply device (60) is connected by the Teflon (RTM) tube to the interval gas port set before the public channel inflow. When previous gas sample has been collected, the control system sends a signal to a solenoid valve or sensor to open the interval gas channel inputting gas according to a set flow rate and timing. The interval gas flow rate, timing and start-stop are also manually controlled by swirling the buttons and switches of the manual flow control valve. The next sample is detected at the end of the interval gas and is cycled until all samples have been detected. The addition of interval gas to achieve continuous detection of multiple gas samples while delaying the diffiision of gas samples and preventing cross-interference between them as much as possible. While the interval gas is detected together with gas samples that can be observed in real time cross-interference, gas sample diffusion and channel pollution to adjust the experiment in time and to ensure the efficiency of the experiment. Each units of the device are independent, and can be freely combined, that is, through different combinations to meet different technical needs applicable to a variety of ways to collect gas samples. As shown in Figure 6, the combination of interval gas unit, gas inlet diversion unit, multichannel gas path automatic switching unit, expiratory chamber unit of mice, and control unit can be used for rapid collection and detection of a number of live exhalation samples, such as mouse exhaled breath or cell headspace. As shown in Figure 7, the position of interval gas supply device (60) is flexible and can be connected to multi-channel gas path automatic switching unit (30), or at the front end of the sample detection channel (400) or the public channel, or can be set up for any diffusion purpose where interval gas is required or where diffusion of gas samples needs to be delayed.
The choice of interval gas has certain requirements and principles. The purpose of the interval gas is to delay the diffusion of gas samples during continuous detection of multiple gas samples, so that there is no interference between neighboring samples as far as possible.
In order to observe the cross-interference status of gas samples, the diffusion degree of gas samples and channel pollution in real time, the selection of interval gas must meet the following criteria: (1) the main components or concentrations of the interval gas are known and can be detected by the testing instrument; (2) the interval gas has less interference to the experimental background and does not react with the substance in the gas sample; (3) The composition of the interval gas is adjusted according to the different gas samples, so as to select the appropriate gas of physical properties (e.g. density, air pressure, concentration, etc.) to delay the diffusion of the gas sample and preventing cross-interference between them as much as possible.
Based on experimental results of different interval gases to achieve continuous detection of multiple gas samples, there are following findings: (1) for continuous detection of multiple biological exhalation samples (e.g. human exhaled breath, cell headspace, plant headspace, etc.), the components of the interval gas can be selected are carbon dioxide (CO2), oxygen (02), high purity nitrogen (N2), dry air, zero gas or inert gas; a mixture of gases formed from two or more of the above gases. Carbon dioxide (CO2) is a common composition of biological exhalation; when used as interval gas, the content ratio of carbon dioxide in the interval gas should be no less than the content ratio of carbon dioxide in gas sample.
(2) interval gas is mainly selected according to the physical and chemical properties of multiple environment atmospheric samples, in which the delayed diffusion effect and interval effect of inert gases are good. Overall, the interval gas is matched according to the characteristics of the gas sample to be detected.
The combination of interval gas can be, for example: 1) )10% zero gas and 20% carbon dioxide; 2) 50% oxygen and 30% high purity nitrogen and 20% carbon dioxide, or 3) 50% helium and 50% argon, etc. The main technical scheme of the invention is based on a continuous fast sampling and detection device for multi-channel gas samples, or a multi-channel sampling and detection device (MSDD) for short, including a multi-channel gas path automatic switching unit (30), a sample detection channel (400), an interval gas supply device (60) and a gas detection device (40). The interval gas supply device (60) is connected to the multi-channel gas path automatic switching unit (30) or to the sample detection channel (400) near the multi-channel gas path automatic switching unit. Said interval gas contains carbon dioxide and contains at least one of the following: oxygen, nitrogen, dry air, zero gas or inert gases. Further, the content ratio of carbon dioxide in the interval gas is not less than the gas sample to be detected.
This invention reveals a multi-channel sampling and detection device (MSDD) to continuously sample and detect multi-channel gas samples. The typical use of the device includes following steps: 1) a gas sample enters sample detection channel (400) via multi-channel gas path automatic switching unit (30); 2) interval gas supply device (60) generates interval gas; 3) multi-channel gas path automatic switching unit (30) switches to next gas sample; 4) for next gas sample, repeat step 1) to 3), otherwise stop.
The multi-channel sampling and detection device (MSDD) applied to mouse exhaled breath detection also contains multiple expiratory chamber units of mice (20) and multiple front buffer gas chambers (25), as shown in Fig.. The front buffer gas chamber (25) is used to pre-collect samples of mice exhaled breath and is installed at the gas output end of the expiratory chamber unit of mice (20).
In order to collect gas samples, the multi-channel sampling and detection device (MSDD) can be further improved to introduce concentrated gas samples into the front buffer gas chambers (25). Accordingly, pushed out from the front buffer gas chambers (25) are concentrated gas samples, and the interval gas supply device (60) generates concentrated interval gas; and the air pressure of the concentrated interval gas is not less than that of the concentrated gas samples.
Sometimes particle filters (70) are needed to remove disturbing particles; as shown in Fig.9, the multi-channel sampling and detection device (MSDD) includes multiple expiratory chamber units of mice (20) and multiple particle filters (70).
The invention also discloses three methods based on multi-channel sampling and detection device (MSDD) using interval gas to achieve continuous detection of multiple samples.
The first method is shown in Figures 7 and 9, when multiple samples are converged to be detected through a public channel or only one instrument needs to continuously analyze multiple samples at the same time so as to delay the diffusion of gas samples and prevent cross-interference between them as much as possible. Multiple gas samples collected online in real time or offline are connected to the injection port on the multi-channel automatic switching unit via rotary table of the Teflon (RIM) tube. The gas outlet of the rotary table is connected with the inlet of the gas detection device, and the sampling sequence, sampling time, interval gas duration and flow rate and other related parameters are set according to the specific requirements, and then the sampling starts. The sampling starts after a period of interval gas has been fed in order to observe whether there are any target residues in the sample detection channel or detection gas chamber. When the gas sample 1 is detected, a peak pattern that is clearly different from that of the interval gas as shown in Figure 11 can be observed. After the detection of gas sample I is completed, and a period of interval gas is fed according to set parameters. After the interval gas inlet is completed, the channel of gas sample 2 is opened to start sampling. This method uses interval gas to delay the diffusion of gas samples and preventing cross-interference between them as much as possible. In such cycle, multiple gas samples can be continuously detected in real time while ensuring that no cross-interference occurs between the samples.
The second method is related to some biological exhalation samples with low target concentration; the diffusion of gas samples during qualitative testing results in some target objects below the instrument's detection limit and thus loses the target, such as human exhaled breath, cell headspace, plant headspace, etc. At present, many gas detections instruments (e.g. high-resolution mass spectrometers) have a short analysis time and can carry out efficient analysis of gas samples, so delaying the diffusion of gas samples can be used to qualitative analysis of gas samples. The rapid diffusion of gas samples is delayed by the use of interval gas between two consecutive samples. Since both the interval gas and samples are detected online in real time, the degree of diffusion between them can be observed through the analysis results of the instrument, so that the target test results can be better judged during the data analysis process.
The third method is applied to gas samples with lower concentration. To increase the concentration of the gas sample, concentration is performed before the sample detection. Sample concentration can be increased by pressurizing and compressing the volume. The gas sample I after the pressure
S
concentration increased at the same time the volume is smaller as shown in Figure 12. The concentrated gas sample is a high-pressure or high-speed gas that is prone to rapid diffiision. Without intervention, it will offset the previous work of pressure and concentration of the sample to increase the concentration. How to achieve real-time continuous detection of concentrated gas samples while delaying the diffusion of gas samples and preventing cross-interference between them as much as possible, some changes can be made in the first method. Adjust the pressure or flow rate of the interval gas according to the pressure of the concentrated gas sample that required to be greater than or equal to the pressure of the concentrated gas sample, as shown in Figure 13. When the pressure of the concentrated gas sample is lower than that of the concentrated gas sample, the two samples will break through the range of the gas sample and cross-interference will occur because the concentrated gas sample will rapidly expand in the sample detection channel. When the pressure of the concentrated interval gas is greater than or equal to the pressure of the concentrated gas sample, it can prevent the concentrated gas sample from diffusing quickly, so that multiple high-pressure concentrated gas samples can be detected continuously and quickly in real time while delaying the diffusion of gas samples and preventing cross-interference between them as much as possible.
The flow rate, pressure and intake time of the interval gas need to match the flow rate, pressure and diffusion rate of the gas sample to make corresponding changes due to the diffusion effect of the gas, and usually the interval gas in the sample time of 60-200 seconds can achieve the separation of the gas sample to prevent cross-interference between them as much as possible.
Brief description of drawings
Figure 1 shows a basic multi-channel sampling device; Figure 2 shows two mouse exhalation chambers and specific gas flow direction; Figure 3 is the schematic diagram of disc-shaped equidistant gas inlet diversion chamber; Figure 4 is a schematic diagram of rotary multi-channel gas path conversion device; Figure 5 is the schematic diagram of conventional gas sample collection process; Figure 6 shows the multi-channel sampling and detection device for animal exhalation; Figure 7 shows the schematic diagram of straight-through multi-channel sampling device; Figure 8 shows multi-channel gas sample sampling and detection device with front buffer gas chamber; Figure 9 is a multi-channel sampling and detection device with particle filters; Figure 10 shows multi-channel sampling method with interval gas; Figure 11 is the schematic diagram of continuous detection results of multiple gas samples; Figure 12 is the comparison diagram of gas sample and concentrated gas sample; Figure 13 is a schematic diagram of continuous detection results of multiple concentrated gas samples; Figure 14 is the schematic diagram of the specific process of parallel sampling of two sets of multi-channel sampling and detection device; Figure 15 is the schematic diagram of the data results of parallel sampling of multi-channel sampling and detection device.
In these figures: 10: gas inlet diversion unit; 20: the expiratory chamber unit of mice; 30: multichannel gas path switching unit; 40: gas detection device; 400: Sample detection channel; 60: Interval gas supply device; 70: particle filter, 25: front buffer gas chamber.
Embodiments In the following, various embodiments of the present invention will be described in detail with reference to the accompanying drawings, and various components can be further designed on the basis of the aforementioned methods.
Embodiment one: selection and matching method of interval gas composition for continuous sampling and detection of biological exhalation samples The main components of different biological exhalation samples are roughly the same, including nitrogen (N2), oxygen (02), carbon dioxide (CO2), water vapor or other gases and impurities. At present, the analysis of biological exhalation samples is mainly to screen out characteristic volatile compounds related to diseases, drugs, environmental exposures and other factors from the gas samples.
The purpose of the interval gas is mainly to delay the diffiision of gas samples during continuous detection of multiple gas samples, so that there is no interference between the samples as far as possible. In order to observe the cross-interference status of gas samples, the diffusion degree of gas samples and channel pollution in real time. The selection of interval gas must meet the following points: ( I) the main components or concentrations of the interval gas are known and can be detected by the testing instrument; (2) the interval gas has less interference to the experimental background and does not react with the substance in the gas sample; (3) The composition of the interval gas is adjusted according to the different gas samples, so as to select the appropriate gas of physical properties (e.g. density, air pressure, concentration, etc.) to delay the diffusion of the gas sample and preventing cross-interference between them as much as possible. Based on the current use of interval gas to achieve continuous detection of multiple gas samples experimental results: The components of the interval gas can be selected carbon dioxide (CO2), oxygen (02), high purity nitrogen (N2), dry air, zero gas or inert gas, such as one or more of the above gases to form a mixture of gases. Carbon dioxide (CO2) as a biological exhalation sample analysis of the common composition of the interval gas, its content ratio of the interval gas generally requires no less than the content ratio of carbon dioxide in the biological exhalation sample.
The following is an example of multiple real-time online mouse exhaled breath samples to continuously detect the component ratio of interval gas. The real-time online collection of mouse exhaled breath samples is mainly through the carrier gas into the detection device, the concentration is lower, and the pressure is lower In order to delay the diffusion of the mouse exhaled breath sample, attention should be paid to the choice of interval air with a higher density and a relatively higher air pressure. In this example, carbon dioxide (CO2) and dry air are selected, and the ratio is 20% carbon dioxide (C07) and 80% di-y clean air, with a flow rate of IL/min and a passage time of 30s. The results show that there is no cross-interference between all samples, and the diffusion of gas samples is effectively delayed.
Embodiment two: selection and matching method of gas composition at intervals for continuous sampling and detection of environmental atmosphere samples The composition of environmental atmospheric samples is more complicated than that of biological exhalation samples. The difference between the concentration and pressure of different environmental atmospheric samples is also large, and some secondary reactions are prone to occur.
When choosing the composition of the interval gas, pay attention to the chemical and physical properties of the compound that are stable and not easy to react with substances in the environmental atmosphere sample. The test shows that the inert gas has a better delaying diffusion effect and spacing effect on the ambient atmosphere, and argon has the best delaying effect. The composition matching method of the interval gas for continuous detection of multiple environmental atmospheric samples can be selected according to the physical and chemical properties of the gas sample. The current test selects 100% argon and 50% argon and 50% helium as the environmental monitoring station. The interval gas for continuous detection of environmental atmospheric samples at a certain site at multiple time points. The results show that both solutions can effectively delay the diffusion of gas samples and prevent cross-interference between gas samples.
Embodiment three: the workflow of the interval gas in the continuous and rapid sampling and detection device of the straight-through multi-channel gas sample The straight-through multi-channel gas sample continuous and rapid sampling and detection device and method are suitable for gas samples with higher concentrations, such as environmental atmospheric samples. Detailed description of the invention as shown in Figure 9 and Figure 11, certain small changes are made to the schematic diagram of Figure 9 to cancel the gas inlet diversion unit, the expiratory chamber unit of mice and the exhaust gas discharge pipeline, when multiple samples converged to be detected through a public channel or only one sample of the instrument needs to continuously analyze multiple samples at the same time need to delay the diffusion of gas samples and preventing cross-interference between them as much as possible. Multiple gas samples collected online in real time or offline are connected to the injection port on the multi-channel automatic switching unit rotary table of the Teflon (RTM) tube, and set the sampling sequence and injection according to specific requirements. The gas outlet of the rotary table is connected with the inlet of the gas detection device, and the sampling sequence, sampling time, interval gas duration and flow rate and other related parameters are set according to the specific requirements, and then the sampling starts. The sample detection begins to advance for a period of interval gas to observe whether there are any target residues in the sample detection channel and detection gas chamber. When the gas sample 1 is detected, a peak pattern that is clearly different from that the interval gas as shown in Figure 11 can be observed. After the detection of gas sample 1 is completed, and a period of interval gas is entered according to the set parameters. After the interval gas inlet is completed, the channel of gas sample 2 is opened to start sampling. This method uses interval gas to delay the diffusion of gas samples and preventing cross-interference between them as much as possible, In this cycle, multiple gas samples can be continuously detected in real time while ensuring that no cross-interference occurs between the samples.
Embodiment four: the workflow of the interval gas in of multi-channel gas samples with front buffer gas chamber for continuous and rapid sampling and detection device.
For biological exhalation samples such as animal exhaled breath, human exhaled breath, and cell headspace, these sample concentrations may be lower than the detection limit of the gas detection equipment. To increase the concentration of the gas sample, the concentration is performed before the sample detection. For example, the sample concentration can be increased by pressurizing and compressing the volume.
The following is an example of multiple real-time online mouse exhaled breath samples to continuously detect the component ratio of interval gas. The gas inlet diversion unit is connected to the mouse expiratory chamber unit through a Teflon (RIM) tube, and the other end is connected to a carrier gas supply device that meets the experimental requirements for the mouse to breathe. During the collection of the target mouse exhaled breath sample, the other mouse exhaled breath channel is in an empty state (the carrier gas and mouse exhaled breath are discharged from the exhaust port), and the carrier gas will be continuously provided to the mouse to breathe to maintain life. A front buffer gas chamber (70) is provided between the expiratory chamber unit of mice and the multi-channel gas path automatic switching unit for pre-collecting a certain volume of mouse exhaled breath samples.
The multi-channel gas path automatic switching unit is equipped with a rotary table, an electronic control module and a high-precision mass flow controller which can accurately control the gas collection flow rate. The multi-channel sampling control program software sets parameters such as carrier gas flow rate, sampling sequence, sampling time and flow rate to meet sampling requirements. The electric control module guides the rotation of the rotary table to switch the gas path and connects the target sample collection path to the sample detection channel (400) of the gas detection device (40) for real-time online analysis.
When the post piston of the buffer gas chamber of the mouse exhaled breath sample I will quickly push out the gas in the cavity to complete the pressure and concentration of the mouse exhaled breath sample to increase the sample concentration and concentrate the mouse exhaled breath. After the detection of gas sample 1 is completed, and a period of concentrated interval gas that matches the pressure of the concentrated mouse exhaled breath sample (greater than or equal to) is input to prevent the rapid spread of the sample. After the inhalation of the concentrated interval gas is completed, the mouse exhaled breath sample 2 is opened to start detection.
Embodiment five: the workflow of two sets of multi-channel sampling and detection One set of multi-channel sampling and detection device has limited gas channels. Multiple sets of multi-channel sampling device and one detection device can be used to continuous and rapid sampling and detection to perform parallel sample testing through sample switching. In this invention, the workflow of two sets of multi-channel sampling device and one detection device are shown in Fig. 14 and Fig. 15. The mechanism of more sets of devices is similar.
Sampling device A and Sampling device B collaborate as follows (shown in Fig.14): 1) device A starts sampling, and the duration takes To; in the meantime, device B is waiting for a duration of Ti; 2) after completion of sampling, device A enters wating stage for a duration of Ti +12; 3) device B starts sampling, for a duration of To; 4) device B enters wating stage for a duration of T2; 5) repeat steps 1) to 4) or stop.
Preferably TI-To +12 Both device A and device B are connected to the same detection device (40).
Embodiment six: application of multi-channel sampling and detection method in cell headspace Breath analysis is currently also used in the analysis of cell gas cultured in vitro, but the current analysis of volatile organic compounds (VOCs) in the cell gas is performed on a single cell bottle. For example, Brunner et al, used component analysis of the cell headspace to identify cancerous and noncancerous cell lines (Discrimination of cancerous and non-cancerous cell lines by headspace-analysis with PTR-MS, 2010). By continuously injecting carrier gas into the cell culture flask to bring the cell headspace into the proton transfer reaction mass spectrometer for real-time online analysis, the analysis data of VOCs in the cell headspace can be obtained. And use a PTFE sampling bag to collect the headspace generated in the cells within 12 hours for offline analysis. A PTFE sampling bag was used to collect the headspace generated within 12 hours of the cells for offline analysis. But for real-time online or offline analysis of the headspace of a single cell, the sample concentration is low, and individual differences between cells and the stability of the sample are ignored, and it is not suitable for simultaneous sampling of parallel samples.
The application of the multi-channel sampling and detection method in the cell headspace is mainly to solve: I). the concentration of volatile organic compounds in the headspace of cells is low and trace amounts. It is easy to lose part of the target during analysis because when the concentration of the target marker is lower than the detection limit, the target marker will not be detected; 2). The different sampling time of the parallel samples of the cell headspace will cause certain interference to the analysis results. For example, the culture time will affect cell proliferation, cell apoptosis, and cvtopathic changes. These factors will have a certain impact on the composition of the cell headspace.
The multi-channel sampling and detection method provided by the present invention can continuously and efficiently analyze a plurality of cell headspace samples collected offline. Multiple parallel samples can be collected at the same time and continuously analyzed, the stability of the parallel samples and the individual differences of the samples can be investigated, and the influence of different cell headspace sampling time and storage time on the experimental data can be excluded.
Claims (4)
- Claims 1 a multi-channel sampling and detection device, said multi-channel sampling and detection device includes two multi-channel sampling devices and one gas detection device (40); each multichannel sampling device includes a multi-channel gas path automatic switching unit (30), a sample detection channel (400), an interval gas supply device (60); said interval gas supply device (60) is connected to said multi-channel gas path automatic switching unit (30) or to said sample detection channel (400); interval gas supplied by said interval gas supply device (60) contains carbon dioxide and contains at least one of the following gases: oxygen, nitrogen, dry air, zero gas or inert gases; wherein, each multi-channel sampling device includes multiple expiratory chamber units of mice (20) and multiple front buffer gas chambers (25); each said front buffer gas chamber (25) is installed at gas output end of one of said expiratory chamber units of mice (20) to pre-collect samples of mice exhaled breath.
- 2 a device according to claim 1, wherein, pushed out from the front buffer gas chambers (25) are concentrated gas samples, and the interval gas supply device (60) generates concentrated interval gas; and the air pressure of said concentrated interval gas is not less than that of said concentrated gas samples.
- 3 a device according to claim 1, wherein, the content ratio of carbon dioxide in said interval gas is chosen to be greater than the content ratio of carbon dioxide in said gas sample.
- 4 a device according to claim 1, wherein, said multi-channel sampling device includes multiple expiratory chamber units of mice (20) and multiple particle filters (70).a device according to any of claims 1 to 4, wherein, said multi-channel gas path automatic switching unit (30) is equipped with a rotary table, an electronic control module and a high-precision mass flow controller.
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