CN111257399A - Direct mass spectrum detection device for high-throughput measurement of volatile organic compounds in blood - Google Patents
Direct mass spectrum detection device for high-throughput measurement of volatile organic compounds in blood Download PDFInfo
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- CN111257399A CN111257399A CN201811458979.1A CN201811458979A CN111257399A CN 111257399 A CN111257399 A CN 111257399A CN 201811458979 A CN201811458979 A CN 201811458979A CN 111257399 A CN111257399 A CN 111257399A
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Abstract
The invention discloses a direct mass spectrometry detection device for high-flux measurement of volatile organic compounds in blood, which comprises a sample bottle for containing sample liquid to be detected, a water bath kettle and a stirring magneton, wherein the sample bottle is arranged in the water bath kettle, and the stirring magneton is arranged in the sample bottle; the sample bottle is matched with a bottle cap with a central hole, a stainless steel gasket is clamped and fixed on the inner side of the bottle cap, and a carrier gas pipe jack and a sample gas pipe jack are arranged on the stainless steel gasket and are both in the central hole range of the bottle cap. One end of the carrier gas pipe is connected with a carrier gas source, the other end of the carrier gas pipe enters the sample bottle, one end of the sample gas pipe is connected with the sample gas pipe jack, and the other end of the sample gas pipe is connected with the mass spectrometer and the tail gas pipe through a tee joint. The method solves the problem that a large amount of foams are easily generated to cause instrument pollution when Volatile Organic Compounds (VOCs) in blood are measured by a bubbling, purging and sample injection mass spectrometry method, the whole operation process is simple and quick, the whole process is less than 10min, and the rapid and high-flux detection of the volatile organic compounds in the blood is realized.
Description
Technical Field
The invention belongs to the technical field of detection of volatile organic compounds in blood, relates to a mass spectrum sample introduction device, and particularly relates to a direct mass spectrum detection device for high-throughput measurement of volatile organic compounds in blood.
Background
Volatile Organic Compounds (VOCs) refer to organic compounds which are easy to volatilize at normal temperature, and are widely known to include formaldehyde, benzene, dichloromethane, trichloroethylene and the like, most of the organic compounds have biological toxicity, and the toxicity is mainly reflected in that the maladjustment of the immune level of an organism can be caused, and the central nervous system and the digestive system are influenced; in severe cases, the liver and hematopoietic system can be damaged. Some VOCs even have carcinogenicity, teratogenicity and mutagenicity, and can induce a plurality of potential chronic diseases, thereby bringing great harm to human health. The evaluation of the VOCs in the human blood is probably closer to the real level of the VOCs in a certain target tissue or certain target tissues on a substrate, so that more complete data can be obtained, therefore, the level of the VOCs in the blood is an important index for evaluating the human health, and has very important significance for analyzing and detecting the VOCs.
Because blood is a complex matrix, when VOCs in whole blood is measured, sample pretreatment is required, and currently, common methods include a headspace method, solid phase extraction, solid phase microextraction, a purging and trapping method, heating and evaporation and the like. These methods have certain limitations: the headspace method is simple, time-saving and easy to operate, but the enrichment effect is not good, and the sensitivity is low; the solid phase extraction method is simple in operation, needs a small amount of solvent, is complicated in steps, is easy to cause analyte loss, and is poor in reproducibility; the solid phase microextraction method has the advantages of high efficiency, small sample destructiveness and the like, can be used for quickly analyzing volatile components in combination with gas chromatography/mass spectrometry (GC/MS), but also has the defects that extraction fibers cannot be repeatedly used, the experiment cost is high, the instrument popularization rate is low, and the method is not suitable for large-scale biological sample detection; because the blood contains a large amount of protein components, a large amount of foam can be generated by a sweeping and trapping method, so that the instrument is easy to pollute, and although the problem that the sample is easy to generate foam can be solved by adding the defoaming agent into the blood sample, the sample is easy to pollute; heating to evaporate into a gaseous form may change the original state of the sample, destroying the sample structure. Therefore, it is very important to develop a direct detection method for detecting volatile organic compounds in blood at high throughput. The high-throughput mass spectrometry technology can directly obtain molecular weight information of a sample, has the advantages of high analysis speed, high sensitivity, good universality and the like, is widely accepted in the field of high-throughput detection, and is developed at a rapid speed.
Disclosure of Invention
The invention aims to provide a direct mass spectrometry detection device for high-flux measurement of volatile organic compounds in blood, which aims to overcome the problem that Volatile Organic Compounds (VOCs) in blood are easily polluted by a large amount of foams generated when the VOCs are measured by a bubbling blowing sample introduction mass spectrometry method. After the analysis is completed, the remaining sample is continuously blown out of the instrument with a purge gas in preparation for the next test cycle. The rapid high-flux analysis and detection of Volatile Organic Compounds (VOCs) in blood are realized.
In order to achieve the purpose, the invention adopts the following technical scheme:
the direct mass spectrometry detection device for high-throughput measurement of volatile organic compounds in blood is characterized by comprising a sample bottle 1 for containing sample liquid to be detected, a water bath 6 and a stirring magneton 7, wherein the sample bottle 1 is arranged in the water bath 6, and the stirring magneton 7 is arranged in the sample bottle 1 and is used for stirring the sample liquid to be detected;
the sample bottle 1 is matched with a bottle cap 5, the center of the bottle cap 5 is provided with a hole, a stainless steel gasket 3 is clamped and fixed on the inner side of the bottle cap 5 with the center provided with the hole, and the opening of the sample bottle 1 is provided with a sealing ring 2 for sealing;
a carrier gas pipe jack is arranged on the stainless steel gasket 3, one end of the carrier gas pipe 10 is connected with a carrier gas source, and the other end of the carrier gas pipe enters the sample bottle 1 through the carrier gas pipe jack arranged on the stainless steel gasket 3; meanwhile, a sample gas pipe jack is arranged on the stainless steel gasket 3, one end of a sample gas pipe 9 is connected with the sample gas pipe jack, and the other end of the sample gas pipe is connected with the mass spectrometer 8 and the tail gas pipe 11 through a tee; the carrier gas pipe jack and the sample gas pipe jack on the stainless steel gasket 3 are both in the central hole opening range of the bottle cap 5.
Furthermore, one end of the sample air pipe 9 is connected with a sample air pipe jack on the stainless steel gasket 3 through the sample air pipe joint 4.
Further, the sample air pipe jack on the steel gasket 3 is in threaded connection with the sample air pipe joint 4.
Further, the carrier gas pipe 10 is connected with a sample inlet of the mass spectrometer 8 through a tee joint, and the mass spectrometer 8 is cleaned by carrier gas purging.
The direct mass spectrometric detection method for high-throughput measurement of volatile organic compounds in blood is characterized by comprising the following steps:
(1) preparing a sample solution to be tested
Taking a blood sample by using a pipette, putting the blood sample into a sample bottle 1, diluting the sample liquid according to the detection requirement, then putting a stirring magneton 7, sealing the sample bottle 1, and sealing and shaking the carrier gas pipe jack and the sample gas pipe jack on the stainless steel gasket 3 uniformly by using a sealing strip;
(2) incubation
Putting the sample bottle 1 into a constant-temperature water bath 6 at 37 ℃, stirring the sample liquid to be detected by the stirring magnetons 7, and starting an incubation process, wherein the incubation time is 0-5 min; the purpose of incubation was two: firstly, because the incubation temperature is close to the body temperature, the sample is in a milder condition, the original state of the sample cannot be changed, and the structure of the sample cannot be damaged; secondly, an enrichment effect is achieved, in the incubation process, not only is the sample slowly heated, but also the stirring magneton 7 is added for stirring, so that as many substances as possible are volatilized and detected;
(3) sample introduction
After incubation, connecting one end of the carrier gas pipe 10 with a carrier gas source, and enabling the other end of the carrier gas pipe to enter the upper part of the sample liquid to be detected in the sample bottle 1 through a carrier gas pipe jack arranged on the stainless steel gasket 3; carrier gas is used as purge gas to purge the liquid surface of the sample liquid to be detected, organic matters on the liquid surface are continuously updated and are carried into headspace gas of the sample bottle 1, one end of a sample gas pipe 9 is connected with a sample gas pipe jack on a stainless steel gasket 3 through a sample gas pipe joint 4, the other end of the sample gas pipe is connected with a sample injection port of a mass spectrometer 8 and a tail gas pipe 11 through a tee joint, the headspace gas of the sample bottle 1 enters the mass spectrometer through the sample gas pipe 9 to be analyzed, and the accumulated acquisition time of a spectrogram is 1-2 min;
(4) cleaning of
And after sample injection analysis is finished, connecting the gas carrying pipe 10 to a sample injection port of the mass spectrometer 8 through a tee joint, and purging for 5-10 min by using purge gas to enter the next cycle operation.
Further, the purge gas is clean air.
Compared with the prior art, the invention has the beneficial effects that:
the invention uses headspace blowing for sample injection and uses the traditional bubbling method, thereby effectively solving the technical problem that the traditional bubbling method is easy to generate a large amount of foam to cause instrument pollution because blood contains a large amount of protein components; simultaneously, utilize the water bath pot to wait that the blood sample that awaits measuring is incubated in the water bath for 37 ℃ of temperature, make the temperature be close to body temperature, can not change the pristine condition of sample, more can not destroy the sample structure, the process of incubating all uses stirring magneton stirring, makes more organic matters in the blood volatilize and is detected by the analysis. The device has the advantages of simple structure, low cost and easy realization, and can provide balance gas for the sample introduction of the mass spectrometer and ensure that the mass spectrometer works under stable optimal air pressure. The method has simple and rapid whole operation process which is less than 10min, and can realize rapid and high-flux detection of volatile organic compounds in blood.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments described in the present application, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic diagram of a sampling process of the direct mass spectrometry detection apparatus for high-throughput measurement of volatile organic compounds in blood according to the present invention;
FIG. 2 is a schematic view of a sampling process of a direct mass spectrometry detection device for high-throughput measurement of volatile organic compounds in blood;
FIG. 3 is an enlarged view of a portion of FIG. 2 at A;
FIG. 4 is a comparison spectrum of VOCs in blood measured by using a direct mass spectrometry detection device for high-throughput measurement of volatile organic compounds in blood and a bubbling method in example 1 of the present invention;
in the figure: 1-bottle body, 2-sealing ring, 3-stainless steel gasket, 4-sample gas pipe joint, 5-bottle cap, 6-water bath, 7-stirring magneton, 8-mass spectrometer, 9-sample gas pipe, 10-carrier gas pipe and 11-tail gas pipe.
Detailed Description
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict. The present invention will be described in detail below with reference to the embodiments with reference to the attached drawings.
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the invention, its application, or uses. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
The relative arrangement of the components and steps, the numerical expressions and numerical values set forth in these embodiments do not limit the scope of the present invention unless specifically stated otherwise. Meanwhile, it should be understood that the sizes of the respective portions shown in the drawings are not drawn in an actual proportional relationship for the convenience of description. Techniques, methods, and apparatus known to those of ordinary skill in the relevant art may not be discussed in detail but are intended to be part of the specification where appropriate. Any specific values in all examples shown and discussed herein are to be construed as exemplary only and not as limiting. Thus, other examples of the exemplary embodiments may have different values. It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, further discussion thereof is not required in subsequent figures.
In the description of the present invention, it is to be understood that the orientation or positional relationship indicated by the directional terms such as "front, rear, upper, lower, left, right", "lateral, vertical, horizontal" and "top, bottom", etc., are generally based on the orientation or positional relationship shown in the drawings, and are used for convenience of description and simplicity of description only, and in the absence of any contrary indication, these directional terms are not intended to indicate and imply that the device or element so referred to must have a particular orientation or be constructed and operated in a particular orientation, and therefore should not be considered as limiting the scope of the present invention: the terms "inner and outer" refer to the inner and outer relative to the profile of the respective component itself.
Spatially relative terms, such as "above … …," "above … …," "above … …," "above," and the like, may be used herein for ease of description to describe one device or feature's spatial relationship to another device or feature as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if a device in the figures is turned over, devices described as "above" or "on" other devices or configurations would then be oriented "below" or "under" the other devices or configurations. Thus, the exemplary term "above … …" can include both an orientation of "above … …" and "below … …". The device may be otherwise variously oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly.
It should be noted that the terms "first", "second", and the like are used to define the components, and are only used for convenience of distinguishing the corresponding components, and the terms have no special meanings unless otherwise stated, and therefore, the scope of the present invention should not be construed as being limited.
Example 1
As shown in fig. 2, the direct mass spectrometry detection device for high-throughput measurement of volatile organic compounds in blood comprises a sample bottle 1 for containing a sample liquid to be measured, a water bath 6 and a stirring magnet 7, wherein the volume of the sample bottle 1 is 60ml, the outer diameter of the bottle body is 24mm, the length of the bottle body is 140mm, the sample bottle 1 is placed in the water bath 6, and the stirring magnet 7 is placed in the sample bottle 1 for stirring the sample liquid to be measured;
the sample bottle 1 is matched with a bottle cap 5, the sample bottle 1 is in threaded connection with the bottle cap 5, a hole is formed in the center of the bottle cap 5, a stainless steel gasket 3 is clamped and fixed on the inner side of the bottle cap 5 with the hole, and a sealing ring 2 with a sealing function is arranged at the bottle opening of the sample bottle 1;
the stainless steel gasket 3 is provided with a gas carrying pipe insertion hole, the gas carrying pipe 10 is a stainless steel pipe with the outer diameter of 3mm and the inner diameter of 2mm, the total length is 16mm, the length of one side of the gas carrying pipe 10 exposed out of the sample bottle 1 is 4mm, the gas carrying pipe is directly connected with a tetrafluoride pipe with the outer diameter of 4mm, the tetrafluoride pipe is connected with clean air serving as a gas carrying source, the length of the gas carrying pipe 10 entering the sample bottle 1 is 12mm, and the gas carrying pipe is arranged above the liquid level of a blood sample to be detected; meanwhile, a sample gas pipe jack with the aperture of 4mm is arranged on the stainless steel gasket 3, the sample gas pipe jack is in threaded connection with a sample gas pipe joint 4 with the inner diameter of 3-5 mm, the sample gas pipe 9 is a tetrafluoride pipe with the outer diameter of 4mm, one end of the sample gas pipe 9 is inserted into the sample gas pipe joint 4, and the other end of the sample gas pipe 9 is connected with a sample inlet of a mass spectrometer 8 and an exhaust pipe 11 through a tee joint; the carrier gas pipe jack and the sample gas pipe jack on the stainless steel gasket 3 are both in the central hole opening range of the bottle cap 5.
After completing a mass spectrum collection, the carrier gas pipe 10 is connected with the sample inlet of the mass spectrometer 8 through a tee joint, and the mass spectrometer 8 is cleaned by carrier gas purging.
The method for detecting by applying the direct mass spectrometry detection device for high-throughput measurement of volatile organic compounds in blood comprises the following steps:
(1) preparing a sample solution to be tested
In the embodiment, VOCs in whole blood is tested, as shown in FIG. 1, a pipette is used for taking 1ml of a whole blood sample and placing the whole blood sample into a sample bottle 1, then 3ml of purified water is taken for diluting the blood sample, a stirring magneton 7 is placed, the sample bottle 1 is sealed, and a carrier gas pipe jack on a stainless steel gasket 3 and a sample gas pipe jack are sealed and shaken uniformly by a sealing strip;
(2) incubation
Putting the sample bottle 1 into a constant-temperature water bath 6 at 37 ℃, stirring the sample solution to be detected by the stirring magneton 7, and starting an incubation process, wherein the incubation time is 2 min; the purpose of incubation was two: firstly, because the incubation temperature is close to the body temperature, the sample is in a milder condition, the original state of the sample cannot be changed, and the structure of the sample cannot be damaged; secondly, an enrichment effect is achieved, in the incubation process, not only is the sample slowly heated, but also the stirring magneton 7 is added for stirring, so that as many substances as possible are volatilized and detected;
(3) sample introduction
After incubation, one end of the gas carrying pipe 10 is communicated with clean air, and the other end of the gas carrying pipe enters the upper part of the sample liquid to be detected in the sample bottle 1 through a gas carrying pipe jack arranged on the stainless steel gasket 3; the method comprises the following steps that clean air is used as purge gas to purge the liquid surface of a sample liquid to be detected, organic matters on the liquid surface are continuously updated and carried into headspace gas of a sample bottle 1, one end of a sample gas pipe 9 is connected with a sample gas pipe jack on a stainless steel gasket 3 through a sample gas pipe joint 4, the other end of the sample gas pipe is connected with a sample injection port of a mass spectrometer 8 and a tail gas pipe 11 through a tee joint, the headspace gas of the sample bottle 1 enters the mass spectrometer through the sample gas pipe 9 to be analyzed, and the accumulated acquisition time of a spectrogram is 2 min;
(4) cleaning of
After sample injection analysis is completed, the carrier gas pipe 10 is connected to a sample injection port of the mass spectrometer 8 through a tee joint, and the next cycle operation can be performed after purging is performed for 5min by using purge gas.
Under the same experimental conditions, the bubbling method and the device and method of the invention are respectively used for testing VOCs in whole blood, namely the experimental conditions are as follows: the volume of the sample bottle 1 is 60ml, the outer diameter of the body of the sample bottle 1 is 24mm, the length of the body of the sample bottle 1 is 140mm, 3ml of purified water and 1ml of whole blood are taken to be placed in the sample bottle 1, the sample bottle 1 is incubated for 2min in water bath at 37 ℃, the flow rate of purge gas is 100ml-min, the accumulated acquisition time of a spectrogram is 2min, and the obtained spectrogram is shown in figure 3.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (6)
1. The direct mass spectrometry detection device for high-throughput measurement of volatile organic compounds in blood is characterized by comprising a sample bottle (1) for containing sample liquid to be detected, a water bath (6) and a stirring magneton (7), wherein the sample bottle (1) is arranged in the water bath (6), and the stirring magneton (7) is arranged in the sample bottle (1) and is used for stirring the sample liquid to be detected;
the sample bottle (1) is matched with a bottle cap (5), a central hole of the bottle cap (5) is formed, a stainless steel gasket (3) is clamped and fixed on the inner side of the bottle cap (5) with the central hole, and a bottle opening of the sample bottle (1) is provided with a sealing ring (2) with a sealing function;
a carrier gas pipe jack is arranged on the stainless steel gasket (3), one end of the carrier gas pipe (10) is connected with a carrier gas source, and the other end of the carrier gas pipe (10) enters the sample bottle (1) through the carrier gas pipe jack arranged on the stainless steel gasket (3); meanwhile, a sample air pipe jack is arranged on the stainless steel gasket (3), one end of a sample air pipe (9) is connected with the sample air pipe jack, and the other end of the sample air pipe (9) is connected with a mass spectrometer (8) and a tail gas pipe (11) through a tee; the carrier gas pipe jack and the sample gas pipe jack on the stainless steel gasket (3) are both in the central hole opening range of the bottle cap (5).
2. The direct mass spectrometry detection device for high throughput measurement of volatile organic compounds in blood according to claim 1, wherein one end of the sample gas tube (9) is connected to the sample gas tube insertion hole on the stainless steel gasket (3) through the sample gas tube connector (4).
3. The direct mass spectrometry detection device for high-throughput measurement of volatile organic compounds in blood according to claim 2, wherein the sample gas pipe jack on the stainless steel gasket (3) is in threaded connection with the sample gas pipe joint (4).
4. The direct mass spectrometry detection device for high-throughput measurement of volatile organic compounds in blood according to claim 1, wherein the carrier gas pipe (10) is connected to the sample inlet of the mass spectrometer (8) through a tee joint, and the mass spectrometer (8) is purged and cleaned by carrier gas.
5. The direct mass spectrometric detection method for high-throughput measurement of volatile organic compounds in blood is characterized by comprising the following steps:
(1) preparing a sample solution to be tested
Taking a blood sample by using a pipette, putting the blood sample into a sample bottle (1), diluting the sample liquid according to the detection requirement, then putting a stirring magneton (7), sealing the sample bottle (1), and sealing and shaking the carrier gas pipe jack and the sample gas pipe jack on the stainless steel gasket (3) by using a sealing strip;
(2) incubation
Putting the sample bottle (1) into a 37 ℃ constant-temperature water bath (6), stirring the sample liquid to be detected by the stirring magneton (7), and starting an incubation process, wherein the incubation time is 0-5 min;
(3) sample introduction
After incubation, connecting one end of a carrier gas pipe (10) with a carrier gas source, and enabling the other end of the carrier gas pipe to enter the upper part of the sample liquid to be detected in the sample bottle (1) through a carrier gas pipe jack arranged on the stainless steel gasket (3); carrier gas is used as purge gas to purge the liquid surface of the sample liquid to be detected, organic matters on the liquid surface are continuously updated and carried into headspace gas of a sample bottle (1), one end of a sample gas pipe (9) is connected with a sample gas pipe jack on a stainless steel gasket (3) through a sample gas pipe joint (4), the other end of the sample gas pipe is connected with a sample injection port and a tail gas pipe (11) of a mass spectrometer (8) through a tee joint, the headspace gas of the sample bottle (1) enters the mass spectrometer through the sample gas pipe (9) to be analyzed, and the accumulated acquisition time of a spectrogram is 1-2 min;
(4) cleaning of
And after sample injection analysis is finished, connecting the gas carrying pipe (10) to a sample injection port of the mass spectrometer (8) through a tee joint, and blowing for 5-10 min by using blowing gas to enter the next cycle operation.
6. The method of claim 5, wherein the sweep gas is clean air.
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| CN112108093A (en) * | 2020-09-28 | 2020-12-22 | 张梦薇 | Device and method for efficiently reacting and extracting generated gas |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112108093A (en) * | 2020-09-28 | 2020-12-22 | 张梦薇 | Device and method for efficiently reacting and extracting generated gas |
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