JP6967198B2 - Breath mist collection device, breath mist analysis system, breath mist collection method and breath mist analysis method - Google Patents

Description

The present invention is for analyzing an exhaled mist collecting device that collects exhaled mist containing a non-volatile substance released from blood or the like via an alveoli, and an exhaled mist collected by the exhaled mist collecting device. It relates to an analysis system including an exhaled mist analysis system, an exhaled mist collecting method and an exhaled mist analysis method, that is, a device and a system for non-invasively collecting and analyzing the non-volatile substance in circulating blood.

Although the dosage and administration of therapeutic drugs are specified, the effective blood concentration range is narrow and the toxicity onset concentration is close to each other. For drugs that fluctuate significantly, therapeutic drug monitoring (TDM) is performed to maintain their effective blood levels. The importance of TDM is also increasing in cancer treatment. In cancer treatment, conventional anti-cancer drugs are also highly toxic to normal cells, so the dose is set so that the anti-cancer effect is maximum and the side effects are not lethal, and the side effects are prepared. The treatment above has been done. However, in recent years, molecular-targeted drugs that act specifically on cancer cells as compared with conventional anticancer drugs have been developed and used for treatment. However, treatment with a molecular-targeted drug is required to maintain its blood concentration between the minimum concentration at which anticancer activity is obtained and the maximum concentration at which side effects do not occur. The blood concentration of a molecular-targeted drug also varies depending on the drug-metabolizing ability of the patient, and the relationship between the dose of the molecular-targeted drug and the blood concentration differs from patient to patient. Therefore, TDM is indispensable for effective treatment. In such a TDM, a measurement sample has been conventionally collected by an invasive method involving blood sampling. Blood sampling imposes a heavy physical and psychological burden not only on patients but also on medical personnel. In addition, blood sampling could not be performed many times a day, and continuous monitoring of blood concentration was not possible. Furthermore, a certain amount of blood sample is required for TDM, and it is impossible for the patient to collect blood at home.

Under these circumstances, there has been a great demand for non-invasive methods to replace invasive methods such as blood sampling for both patients and medical personnel. Biosampling by non-invasive methods that can be performed anywhere and at any time, even if it is not related to medical treatment, is prohibited not only for therapeutic drugs but also for identifying the causative compounds of poisoning by harmful substances, and for banning narcotics and stimulants. It is also eagerly awaited in fields such as confirmation of the presence or absence of drug administration.

Here, as a non-invasive method, a method of analyzing the components contained in the exhaled breath and inspecting the condition of the subject is being studied. For example, Non-Patent Document 1 discloses a relationship between a volatile component released from exhaled breath and health and illness, collection of exhaled breath as an analysis sample, and a method for analyzing the component.

The above-mentioned technique focuses on volatile components such as hydrogen, carbon monoxide, nitric oxide, and volatile organic compounds released in the exhaled breath, but in recent years, non-volatile components have also been attracting attention.

Non-volatile components contained in blood and lung tissue, for example, harmful compounds contained in therapeutic drugs, foods, etc., physiological substances such as hormones, substances that are indicators of diseases such as glucose and lipids, etc. , It is taken up by the mist of alveolar coating fluid (breath mist) consisting of pulmonary surfactant and body fluid formed with respiration and released into the exhaled breath.

In order to analyze such non-volatile components in the exhaled mist, for example, Patent Document 1 discloses a method and an apparatus for collecting an exhaled condensate and measuring a protein, a bioactive factor, glucose, a toxic metal). Has been done.

Further, Patent Document 2 discloses a technique of adhering and capturing an aerosol contained in exhaled breath with a filter and analyzing the aerosol attached to the filter.

Applied Physics Vol. 83 26-32 (2014): Shimouchi, Kondo

Japanese Patent Publication No. 2012-521008 Japanese Patent Publication No. 2014-510914

However, in the technique described in Patent Document 1, there is a problem that the device for collecting the exhaled condensate tends to be large, and the operation is complicated and time-consuming to obtain the condensate. That is, although it is a non-invasive method, it is not a method that anyone can implement at any place and time, and there is a problem that there are many restrictions on implementation.

Further, the technique described in Patent Document 2 is mainly for determining the presence or absence of the use of a prohibited drug, and has a problem that the quantitativeness of the analysis target is poor.

The inventors have stated that the exhaled mist discharged from the alveoli during respiratory movement is an aggregate of fine particles composed of the membrane of pulmonary surfactant existing in the alveoli and the water contained therein. It was noted that the fine particles contained non-volatile substances that infiltrated the alveoli from the circulating blood. That is, they have found that by collecting fine particles composed of pulmonary surfactant membrane and water, non-volatile substances discharged from blood can be efficiently collected and concentrated. As a result, therapeutic drug monitoring (TDM), etc., monitors the concentrations of compounds / substances in the blood and tissues and applies them to the diagnosis and treatment of diseases, the state of uptake of harmful substances into the body, and further. It is possible to know whether or not a prohibited drug such as a drug or stimulant is taken. As a means for that purpose, we have completed an exhaled mist collecting device containing an adsorbent capable of capturing the fine particles discharged from the exhaled air. We also found a method for accurately measuring and quantifying compounds and substances contained in the collected exhaled breath mist.

The present invention is a breath mist collecting device that efficiently collects exhaled mist containing a non-volatile substance released from blood or the like via the alveoli, and a breath mist collected by the exhaled mist collecting device with high accuracy. It is an object of the present invention to provide a breath mist analysis system, a breath mist collection method, and a breath mist analysis method for analysis in.

In order to achieve the above object, in the invention according to claim 1, in order to measure the exhaled breath component, an exhaled mist containing a non-volatile and refractory volatile substance released from blood or the like via the alveoli and the respiratory tract is adsorbed. An exhaled mist collecting device that collects with an agent, an adsorbent unit in which an adsorbent is detachably arranged and exhaled air is circulated through the adsorbent to collect exhaled mist, and the exhaled breath of a subject is collected as the adsorbent. The suitability of the breath mist collection condition is determined based on the breath introduction means introduced into the unit, the breath volume measuring means for measuring the breath volume of the subject, and the breath volume measured each time by the breath volume measuring means. The determination means includes, and the determination means uses a technical means of determining the suitability of the exhaled mist collection condition based on a threshold set based on the closing capacity of the subject.

In the invention according to claim 2 , in the breath mist collecting device according to claim 1 , the adsorbent is an adsorbent capable of specifically capturing a target component having fluorescence. Is used.

In the invention according to claim 3 , the breath mist analysis system adsorbs to the breath mist collecting device according to claim 1 or 2 and the adsorbent unit, and analyzes the collected breath mist. A technical means is used that comprises a device and a lung volume fraction measuring device configured to perform subject lung volume fractionation and measure the subject's closing volume.

In the invention according to claim 4 , in the breath mist analysis system according to claim 3 , the technical means that the analyzer is a fluorescence emission analyzer is used.

In the invention according to claim 5 , the breath mist collecting method introduces the breath of a subject into an adsorbent and contains a non-volatile and refractory substance released from blood or the like through the alveoli and the respiratory tract. a step of collecting comprises a step of measuring the amount of exhalation of the subject, and determining the appropriateness of the collecting conditions exhalation mist based on the expiration of each time that is measured, and the breath mist In the step of determining the suitability of the collection conditions, a technical means of determining the suitability of the breath mist collection conditions based on a threshold set based on the closing capacity of the subject is used.

In the invention according to claim 6 , in the breath mist collecting method according to claim 5 , the adsorbent is an adsorbent capable of specifically capturing a target component having fluorescence. Is used.

In the invention according to claim 7 , in the breath mist analysis method, the breath mist collected by the adsorbent is analyzed by the fluorescence emission analysis method by the breath mist collecting method according to claim 6. Use the means.

According to the first aspect of the present invention, the breath of the subject can be introduced into the adsorbent unit by the breath introducing means, and the breath mist can be collected by the adsorbent of the adsorbent unit. The exhaled volume of the subject can be measured by the exhaled breath measuring means, and the suitability of the exhaled mist collection condition can be determined based on the exhaled volume measured by the exhaled breath measuring means by the determining means, so that the exhaled mist can be efficiently used. Can be collected.

In order for a large amount of exhaled mist to be released during exhalation, it is necessary to exhale to a level below the resting expiratory position. The amount of exhaled mist is small. The amount of exhaled mist released increases rapidly as the depth of breathing increases, but repeated calls that reach the amount of residual air impose a heavy load on the subject, which is not desirable. Therefore, call reaches the closing volume, i.e., be performed repeatedly following call closing capacity, since preferred because more breath mist is released, the threshold value set on the basis of the closing capacity of the examiner A larger amount of exhaled mist can be collected by determining the suitability of the exhaled mist collection conditions based on the above.

When an adsorbent capable of specifically capturing a target component having fluorescence is used as an adsorbent as in the invention according to claim 2, a state capable of directly detecting solid phase fluorescence on the surface of the adsorbent is created. The on-resin solid phase fluorescence detection method can be adopted. According to the on-resin solid-phase fluorescence detection method, exhaled breath is introduced into the adsorbent, the target component is adsorbed / concentrated / fixed on the surface of the adsorbent, unnecessary contaminants are washed, and then the target component is adsorbed / fixed on the surface of the adsorbent. It can be easily quantified by irradiating the excitation light having a wavelength corresponding to the target component in the same state and detecting the fluorescence produced by the target component.

As in the invention of claim 3 , the breath mist collecting device according to claim 1 or 2 is combined with the analyzer and the lung volume fraction measuring device to construct a breath mist analysis system. be able to.

Lung weight fraction measuring device, by measurably configure closing volume of a subject, for determining the appropriateness of the collecting conditions exhalation mist, to determine the threshold set on the basis of the closing capacity of the subject Can be done.

By adopting a fluorescence emission analyzer as the analyzer as in the invention according to claim 4, the target component can be analyzed and quantified with high accuracy.

According to the fifth aspect of the present invention, the exhaled air volume of the subject can be measured, and the suitability of the exhaled breath mist collection condition can be determined based on the measured exhaled air volume to collect the exhaled mist. Exhaled mist can be collected efficiently.

By determining the appropriateness of the collecting conditions exhalation mist based on the threshold value set on the basis of the closing capacity of the examiner, it is possible to collect more breath mist, which is preferable.

According to the sixth aspect of the present invention, an on-resin solid phase fluorescence detection method can be performed by adopting a fluorescence emission spectrometry method as an analysis method.

By adopting the fluorescence emission analysis method as the analysis method as in the invention according to claim 7, the target component can be analyzed and quantified with high accuracy.

It is a block diagram which shows the structure of the exhalation mist collection system. It is explanatory drawing of the exhaled breath mist collecting device. FIG. 2A is an external view, and FIG. 2B is an explanatory view showing the internal structure of the breath mist collecting device. It is explanatory drawing of lung volume fraction (A) and closing volume (B). It is a flowchart which shows the exhalation mist collection method. It is explanatory drawing which shows the synthesis scheme of an adsorbent. It is explanatory drawing which shows the relationship between the drug concentration detected from the exhaled breath mist and the drug concentration in blood. It is explanatory drawing which shows the synthesis scheme of an adsorbent.

(Breathing mist analysis system)
As shown in FIG. 1, the breath mist analysis system S of the present invention collects breath mist containing non-volatile and refractory substances released from blood or the like through the alveoli and respiratory tract with an adsorbent. A collection device 1, an analyzer 2 that analyzes the breath mist collected by the breath mist collection device 1, and a lung volume fraction measurement device 3 that fractionates the lung volume of the subject by a spirogram. I have.

The breath mist collecting device of the present invention collects a breath mist containing a non-volatile and refractory substance released from blood or the like through the alveoli and the respiratory tract with an adsorbent in order to measure the breath component. As shown in FIGS. 1 and 2, the breath mist collecting device 1 arranges the breath introducing means 10 for introducing the breath of the subject into the adsorbent unit 11 and the adsorbent desorbably, and distributes the breath to the adsorbent. It is provided with an adsorbent unit 11 for collecting exhaled breath mist.

Further, the breath mist collecting device 1 determines whether or not the breath mist collection condition is appropriate based on the breath volume measuring means 12 for measuring the breath volume of the subject and the breath volume measured by the breath volume measuring means 12. Means 13 and.

The exhaled breath introducing means 10, the adsorbent unit 11, and the exhaled breath amount measuring means 12 are connected side by side in this order when viewed from the subject side.

The exhaled breath introducing means 10 includes an exhaled breath collecting member 10a such as a mask covering the subject's mouth and a mouthpiece that the subject holds and exhales, and the exhaled breath collecting member 10a so that the subject can repeatedly breathe while wearing the exhaled breath collecting member 10a. The check valve 10b and the like are provided. Here, the inner surface of the exhaled breath recovery member 10a is preferably hydrophobic in order to introduce the exhaled breath mist into the adsorbent without leakage.

The adsorbent unit 11 is arranged so that the cartridge 11a, which is filled with the adsorbent K and is made of a mesh-like material or punching metal which is configured to be breathable, and the cartridge 11a are detachably arranged downstream of the exhaled breath introducing means 10. A member 11b and the like are provided. Here, if the cartridge 11a can be detachably arranged, the arrangement member 11b can adopt various configurations such as sandwiching the cartridge 11a, forming a slit, and inserting the cartridge 11a.

The exhaled breath measuring means 12 is composed of, for example, a flow meter, and is configured to be capable of measuring the exhaled flow rate and the total exhaled volume of the subject. It can also be configured to be equipped with a respiratory rate counter so that the respiratory rate can be measured.

The determination means 13 compares the amount of exhaled air sent from the exhaled air volume measuring means 12 with a threshold value input in advance, and determines whether or not the exhalation mist collection condition is good for the exhalation mist collection. Judge suitability. Here, the determination means 13 is provided with a notification means for encouraging exhalation by voice guidance or the like so that the subject satisfies a good exhalation condition.

It is possible to adopt a configuration in which the exhaled breath introducing means 10 and the adsorbent unit 11 are provided with a removing means such as a filter and a trap mechanism for removing water and impurities in the exhaled air introduced into the adsorbent unit 11. .. According to this, it is possible to prevent the liquid such as saliva of the subject from flowing into the adsorbent and to maintain the sensitivity of the exhaled breath measuring means 12.

As the analyzer 2, an analyzer suitable for the measurement target, such as a known fluorescence emission analyzer, an absorptiometry analyzer, or a liquid chromatography mass spectrometer, can be used. In this embodiment, a fluorescence emission analyzer capable of analyzing and quantifying a target component with high accuracy is adopted.

As the lung volume fraction measuring device 3, a known spirometer 30 for performing lung volume fractionation can be used. Lung volume fractions can be performed based on the spiralograms obtained by the spiral meter 30. In addition, a configuration including a nitrogen analyzer 31 for acquiring a nitrogen exhalation curve can be adopted. This makes it possible to measure the closing volume described later.

(Breathing mist)
In the peripheral structure of the lung, the terminal bronchiole branches into the respiratory bronchiole, and further branches into the alveolar duct and the alveolar sac. The surface of the alveoli is covered with alveolar coating fluid consisting of pulmonary surfactant produced by type II alveolar epithelial cells, intracellular fluid of alveolar macrophages, and interstitial tissue fluid.

The peripheral airways, including the bronchioles, are obstructed by exhalation and open by inspiration. The flow forms an exhaled mist. This exhaled mist is a part of the airway covering liquid that is released into the airway in the form of a mist and then released to the outside by exhalation.

Pulmonary surfactant is a mixture of lipid and protein whose main component is dipalmitoylphosphatidylcholine, which is a lipid phosphate, and acts as a surfactant to prevent air from collapsing from the alveoli. Four types of surface-active substance-related proteins (SP) are known as proteins constituting pulmonary surfactant. SP-A and SP-D are glycoproteins, and SP-B and SP-C are lipid-affinity proteins.

The alveoli sac is a small mass of alveoli where gas exchange with the outside air and blood takes place. The alveolar coverage thinly covers the inner surface of the alveoli and is in contact with blood across the interstitial membrane. Non-volatile and refractory substances contained in blood and lung tissue, for example, harmful compounds contained in therapeutic agents, foods, etc., bioactive substances such as hormones, energy of living organisms such as glucose and lipids, etc. Substances and the like move from the blood into the alveolar coating.

Then, the alveolar covering solution containing the non-volatile and non-volatile drug becomes the bronchial covering solution together with the secretory components from the bronchial line, and is released into the exhaled breath as an exhaled mist. By collecting this exhaled mist with an adsorbent and measuring the above-mentioned compounds / substances contained therein, the pharmacokinetics thereof can be grasped. This makes it possible to monitor the concentrations of the above compounds / substances in blood and tissues, monitor therapeutic drugs, apply them to diagnosis / treatment of diseases, grasp the state of uptake of harmful substances into the body, and the like. ..

(Lung volume fraction)
The amount of exhaled mist released into the exhaled breath varies depending on the depth of breathing. Qualitatively, the peripheral airway is obstructed during deep exhalation, and when breathing is replaced by inspiration, the obstructed peripheral airway is opened, and at that time, the airway epithelial covering fluid is scattered and floats as an aerosol. Deep breathing releases more exhaled mist because it is released during exhalation.

FIG. 3 schematically shows a spirogram, a lung volume fraction, and a nitrogen exhalation curve. Pulmonary volume is the maximum inspiratory level, which is the maximum inhaled breathing level, the maximum expiratory level, which is the maximum exhaled breathing level, the rested inspiratory level, which is the naturally inhaled breathing level, and the natural breathing. It is divided into four levels, the resting and expiratory level, which is the breathing level at which the patient exhales.

In addition, the following air volume is defined by the air volume between the above four levels.
・ Tidal volume: difference between resting inspiratory position and resting expiratory position ・ Preliminary inspiratory volume: difference between maximum inspiratory position and resting inspiratory position ・ Preliminary expiratory volume: difference between maximum expiratory position and resting expiratory position ・ Pulmonary activity: 1 times ventilation, the sum of the pre-intake amount and pre-expiratory volume and functional residual volume: sum-total lung capacity of the preliminary expiratory volume and residual volume: where the sum of the vital capacity and the residual capacity, residual volume Is the amount of air remaining in the lungs after maximal exhalation.

Normal breathing is resting breathing that reciprocates between the resting inspiratory position and the resting expiratory position.

The volume called from the timing of obstruction of the peripheral airway to the time of exhalation is called the closing volume (CV). The sum of the residual air volume and the closing volume is called the closing capacity. The closing volume is obtained by the nitrogen exhalation method or the like. The nitrogen exhalation curve used to measure the closing volume will be described below.

The nitrogen exhalation curve is indicated by the concentration of nitrogen in the exhaled air relative to the air volume. First, inhale as much pure oxygen as possible at the maximum exhalation position to fill the alveoli. Since there is a difference in elevation between the apex of the lung and the bottom of the lung, the pressure at the apex of the lung is originally lower than that of the bottom of the lung, and the alveolar space is large. At the bottom of the lung, the alveolar space shrinks, the peripheral airway obstruction rate is high, and the oxygen filling rate associated with inspiration is high, that is, the nitrogen content is low. In the nitrogen exhalation curve, pure oxygen in the airway is first exhaled in Phase I, and then nitrogen from the bottom of the lung is mixed and exhaled in the airway in Phase II. Phase III is the period during which a gas with a relatively constant nitrogen concentration is exhaled by the exhalation of alveolar air, and the rate of increase in nitrogen is moderate. In Phase IV, the bottom of the lung begins to occlude, nitrogen is exhaled from the apex of the lung, which has been inadequately replaced with pure oxygen, and the nitrogen concentration rises again. This phase IV range is the closing volume.

In order for a large amount of exhaled mist to be released during exhalation, it is necessary to exhale to a resting exhalation level or lower. Since the peripheral airway is less obstructed in the resting expiratory position, the amount of exhaled mist released with the opening of the peripheral airway is small.

According to J Appl Physiol 108: 584-588, 2010, the number of particles with a particle size of 0.30-0.40 μm at the resting expiratory position is 6700, while the number of particles at the closing capacity is 13000. The number of particles at the residual air volume level is 44,000, and the amount released increases rapidly as the depth of breath increases.

Here, it is not desirable to repeat the call to reach the residual air amount because it imposes a heavy load on the subject. Therefore, it is preferable to repeatedly call the closing volume because a larger amount of exhaled mist is released.

(How to collect exhaled mist)
Next, a method of collecting exhaled mist will be described with reference to FIG.

First, in step S1, a subject's spiralogram is acquired by a spirometer 30 by a known method, and lung volume fractionation is performed. In addition, the nitrogen exhalation curve of the subject is acquired using the nitrogen analyzer 31 and the closing volume is measured.

In the following step S2, the resting inspiratory position, the resting expiratory position, the closing volume, and the number of breaths to be performed for the expiratory mist collection are input to the determination means 13 of the expiratory mist collecting device 1.

In the following step S3, the determination means 13 sets the threshold value Vth calculated based on the closing capacity.

In the following step S4, the subject is equipped with the exhaled mist collecting device 1, and the subject introduces deep exhaled breath after several resting breaths. The exhaled breath introduced by the subject is introduced into the adsorbent unit 11, and the exhaled breath mist is collected by the adsorbent K. The exhaled volume introduced by the subject is measured by the expiratory volume measuring means 12, and the expiratory flow rate and the expiratory capacity are sent to the determining means 13.

In the following step S5, the amount of exhaled breath sent from the exhaled breath amount measuring means 12 is compared with the threshold value Vth, and the suitability of the collection condition of whether or not the exhalation condition is good for collecting the exhaled mist is determined. Specifically, it is determined whether or not the exhalation level of the subject has reached the closing capacity based on the expiratory capacity based on the resting inspiratory position or the resting expiratory position.

When the exhalation level of the subject reaches the closing capacity (step S5: YES), the notification means provided by the determination means 13 notifies by voice or the like that the exhalation is terminated, and the process proceeds to step S6.

When the exhalation level of the subject has not reached the closing capacity (step S5: NO), the notification means provided by the determination means 13 notifies the subject to urge exhalation by voice or the like.

In step S6, the number of breaths is counted by the determination means 13.

In the following step S7, it is determined whether or not the breathing rate has reached the set breathing rate. If the set breathing rate has not been reached (step S7: NO), step S4 or lower is repeated, and if the set breathing rate is reached (step S7: YES), the collection of exhaled mist is terminated. ..

Here, the exhalation approaching the closing volume is about 10% of the vital capacity in a healthy person, and a level at which the exhalation is less by 10% of the vital capacity from the maximum expiratory position may be used as a representative value of the closing capacity. More simply, it can be set based on the vital capacity estimated from height, age, gender and the like.

(Adsorbent)
The exhaled mist is considered to be a surface hydrophobic reverse micelle. As the adsorbent, various adsorbents can be used as long as they can sufficiently capture such exhaled mist, but in the present embodiment, the adsorbents are glycidyl methacrylate (GMA) and divinylbenzene (GMA) as shown in FIG. BG15 was used in which the glycidyl group of the polymer obtained by suspension polymerization of DVB) was hydrolyzed with sulfuric acid. The design concept of this adsorbent is to prevent the detachment of water droplets due to the disintegration of micelles on the mist surface after adsorption of the hydrophobic part by appropriately distributing the hydrophilic part derived from the ring-opening diol group and the hydrophobic part derived from the benzene ring. It is in. The adsorbent used in the present invention is not limited to the copolymer of GMA and DVB. For example, ethylene glycol dimethyl acrylate (EGDM) is used as a cross-linking agent instead of DVB, and the ratio with GMA is determined by the above design concept. Examples thereof include a prepared copolymer. Since the exhaled mist is trapped in the adsorbent according to this design concept, all the drugs contained therein are trapped regardless of whether they are hydrophobic or hydrophilic.

(Breathing mist analysis method)
The adsorbent unit 11 that has collected the exhaled mist is removed from the exhaled mist collecting device 1 and subjected to analysis. As the analyzer 2, an analyzer suitable for the measurement target such as a fluorescence emission analyzer, an absorptiometry analyzer, a liquid chromatography / mass spectrometer, etc. can be used.

As the fluorescence emission analysis method, an on-resin solid phase fluorescence detection method can be adopted. In the on-resin solid-phase fluorescence detection method, when an adsorbent capable of specifically capturing the target component having fluorescence is used, the matrix is hardly present on the cartridge filled with the adsorbent, and the surface of the adsorbent is eliminated. It is a detection method that creates a state in which direct solid-phase fluorescence can be detected in (A. Yamamoto, et al., Anal.Methods, 2013, 5, 773-777.). According to this detection method, high sensitivity is achieved by concentrating the target component on the surface of a small amount of solid-phase extractant.

Although the method of the present invention captures the breath mist, it is also possible to impart selectivity to the drug to be analyzed to the adsorbent that captures the breath mist. By using such an adsorbent, it can be directly used in the on-resin solid phase fluorescence detection method. For example, an adsorbent in which a functional group that specifically adsorbs a component to be measured is introduced into a base gel using BG15 can be prepared, and the adsorbent that has adsorbed exhaled breath can be used as it is. Introduce exhaled air into the adsorbent, adsorb, concentrate and fix the target component on the surface of the adsorbent, wash unnecessary contaminants with acetonitrile or medicated ethanol in the case of home, and then adsorb and fix the target component on the surface of the adsorbent. It can be easily quantified by irradiating the excitation light having a wavelength corresponding to the target component in the same state and detecting the fluorescence produced by the target component.

The on-resin solid-phase fluorescence detection method device is easy to miniaturize and portable design, and such a device enables analysis anywhere, that is, on-site or bed-site analysis. Furthermore, by adding a data transmission function to this device or connecting to a data transmission terminal, for example, the patient himself or her caregiver taking a therapeutic drug can collect exhaled mist and quantify the drug at home or the like. It is also possible to do this and automatically send the data to the attending physician. It is also possible for doctors and others to carry out bed site monitoring in this method at medical institutions.

In the breath mist analysis system S, it is preferable that the analyzer 2 has a quantitative value calculation function using a calibration curve. In this case, there is a method of measuring a quantitative object having a known concentration in advance and calculating a calibration curve as an internal standard. If the purpose is achieved by confirming that the substance to be quantified is between the lower limit amount and the upper limit amount, enter only the quantitative values of the standard product of the lower limit amount and the upper limit amount. Is also possible.

The amount of exhaled mist generated varies depending on the amount of exhaust from the lungs. In order to correct this fluctuation, it is possible to measure the amount of substances that can be references such as inorganic ion species and SP-D at the same time, and convert the drug amount ratio to this into the blood concentration. Similar to the drug measurement, a fluorescence emission spectrometer, an absorptiometry analyzer, a liquid chromatography / mass spectrometer, or the like can be used for the measurement of these references. Further, it is also possible to use a measurement method or a chemical analysis method using an antigen-antibody reaction, an enzyme reaction, or the like. Further, the therapeutic drug may be contained in a fixed ratio of a substance that can be quantified by exhalation and has no effect, or it may be co-administered to obtain the above reference. Although it is possible to add a reference measurement function to an on-site device for general households, it is realistic to prioritize portability and small size and respond with the amount of drug per expiratory exhaust volume. When judging by the amount of drug per exhaled breath, it is preferable to standardize the breathing method at the time of collecting exhaled mist.

The feature of this method is that the non-volatile and non-volatile components released in the exhaled breath can be easily collected in a short time. Further, this method captures the breath mist containing the measurement target, not the object to be measured, and even when measuring a plurality of body fluid components, it can be carried out with one kind of breath mist adsorbent.

(Change example)
As the threshold value Vth, a value equal to or lower than the resting expiratory position and greater than or equal to the closing capacity can be adopted.

(Effect of embodiment)
According to the breath mist collecting device 1 and the breath mist collecting method of the present invention, the breath of the subject is introduced into the adsorbent unit 11 by the breath introducing means 10, and the breath mist is collected by the adsorbent K of the adsorbent unit 11. can do. The exhaled mist is made efficient by measuring the exhaled volume of the subject by the exhaled breath measuring means 12 and determining the suitability of the exhaled mist collection condition based on the exhaled volume measured by the exhaled breath measuring means 12 by the determining means 13. Can be collected well.
Further, according to the breath mist analysis system S and the breath mist analysis method, the target component in the collected breath mist can be analyzed and quantified with high accuracy, and can be used for therapeutic drug monitoring and the like.

(Example 1)
(1) Synthetic adsorbent BG15 was used as the adsorbent. The base gel of the adsorbent was synthesized by the suspension polymerization method. First, an organic solvent in which a monomer and a cross-linking agent are dissolved and water are added to a separable flask to form a two-phase flask. The aqueous layer contains a surfactant to prevent coalescence of the obtained oil droplets, and the oil layer also contains a diluent and a polymerization initiator to increase the surface area of the resin. Although the suspension polymerization method was used in this example, the method for synthesizing the adsorbent is not limited to the suspension polymerization method, and for example, a direct membrane emulsification method can also be used. The direct membrane emulsification method is a method in which an oil layer containing a polymerization monomer is directly dispersed in an aqueous layer through a filter having a pore size of several μm.

Next, the liquid is stirred and o / w type oil droplets are suspended in a water tank. After the oil droplet diameter stabilizes, polymerization starts. Warm slowly at 1 ° C / min or less, and when the reaction layer temperature becomes constant at 70-80 ° C, continue the reaction at this temperature for more than half a day. The finished resin was thoroughly washed with Soxhlet or the like, and then spheroidized to obtain an adsorbent having a particle size of about 0.5-1.0 mm required for exhaled breath collection. It is also possible to synthesize a resin having a uniform particle size by the direct membrane emulsification method.

(2) Breath-blood drug concentration correlation experiment The relationship between the drug concentration detected in the breath mist and the drug concentration in the blood was investigated. Subjects took one 60 mg tablet of fexofenadine, a drug administered to hay fever patients as an antiallergic drug on the morning of the experiment, and exhaled breath was collected with blood every two hours.

For the collection of exhaled mist, an adsorbent package having a cell size of φ30 × 10 mm was filled with 1 g of an adsorbent package having a particle size of 0.5-1.0 mm. There was no dyspnea due to pressure loss, and it was possible to collect exhaled breath normally. For exhaled breath, 10 L was collected by resting breathing.

The adsorbent that collected the exhaled breath was eluted with 20 mL of methanol and dissolved in 0.1 mL of water to prepare a test solution, and LC-ESI ⁺ -MS measurement (m / z = 502, detection limit 10 pg / mL). It was offered to.

Blood was measured by diluting it 3-fold with PBS, centrifuging it into plasma, and then subjecting it to LC-UV measurement. Since fexofenadine (FF) has only a benzene ring and does not have a characteristic absorption spectrum, it was detected at UV 220 nm.

The measurement results are shown in FIG. FIG. 6 shows total FF plasma values using formic acid for dilution. A comparison of the breath mist and the blood concentration showed extremely good similarity between them, and it was shown that the blood concentration of the drug can be grasped by collecting and analyzing the breath mist.

(Example 2) Adsorbent for molecular-targeted therapeutic agent In this example, for gefitinib, which is a therapeutic agent for lung cancer, TDM is simplified to determine the optimum dose for each patient using the blood concentration as a surrogate marker. An adsorbent was prepared to realize bedside monitoring. By quantifying the molecular-targeted therapeutic agent collected on the surface of the adsorbent with an on-resin device, it is confirmed that the blood concentration of the molecular-targeted therapeutic agent has reached the effective concentration, and further, the drug efficacy is lost due to forgetting to take. Can be prevented. Bedside monitoring is possible because an on-resin device is used.

For the purpose of evaluating the adsorption selectivity of gefitinib, two types of adsorbents were synthesized, one having a naphthyl group having a conjugated planar structure next to the thiourea group and the other having a bulky cyclohexyl group introduced. FIG. 7 shows the reaction scheme. The base gel used was a polymer in which the ratio of the functional group monomer GMA to the cross-linking agent ethylene glycol dimethacrylate (EGMA) was 30:70, and it was a hydrophilic resin compared to BG15. This glycidyl group was ring-opened with ammonia to form an aminoethanol structure. Naftyl isothiocyanate and cyclohexyl isothiocyanate were introduced via amino groups, respectively, to synthesize Na-ITC and CH-ITC adsorbents. Preliminary tests have confirmed that gefitinib can be supplemented by supporting silver on these adsorbents. Therefore, the capture characteristics of gefitinib were evaluated using a silver-supported ITC adsorbent.

The presence or absence of silver support on two types of adsorbents and the behavior of gefitinib in an acetonitrile solution in a total of four types of 50 mg-filled cartridges were investigated. It was found that the silver-supported Na-ITC adsorbent captures gefitinib in acetonitrile almost quantitatively and can be desorbed with methanol. This adsorbent could be subjected to solid-phase fluorescence analysis by an on-resin apparatus by capturing the exhaled aerosol, washing it with water, and then washing it with acetonitrile. As described above, an adsorbent suitable for the on-resin solid-phase fluorescence detection method of gefitinib could be obtained.

1 ... Breath mist collecting device 2 ... Analytical device 3 ... Lung volume fraction measuring device 10 ... Breath introduction means 10a ... Breath collecting member 10b ... Check valve 11 ... Adsorbent unit 11a ... Cartridge 11b ... Arrangement member 12 ... Breath Volume measuring means 13 ... Judgment means 30 ... Spirometer 31 ... Nitrogen analyzer K ... Adsorbent S ... Breath mist analysis system

Claims (7)

An exhaled mist collecting device that collects exhaled mist containing non-volatile and refractory substances released from blood or the like through the alveoli and respiratory tract with an adsorbent in order to measure the expiratory component.
An adsorbent unit that collects exhaled mist by arranging the adsorbent in a removable manner and circulating exhaled air through the adsorbent.
An exhaled breath introducing means for introducing the exhaled breath of the subject into the adsorbent unit,
An exhalation volume measuring means for measuring the exhalation volume of a subject,
Determination means for determining the appropriateness of the collecting conditions exhalation mist based on the expiration of each time measured by said expiratory volume measuring means,
Equipped with
The determination means is an exhaled mist collecting device, which determines the suitability of an exhaled mist collecting condition based on a threshold value set based on the closing capacity of a subject. The breath mist collecting device according to claim 1 , wherein the adsorbent is an adsorbent capable of specifically capturing a target component having fluorescence. The exhaled mist collecting device according to claim 1 or 2,
An analyzer that adsorbs in the adsorbent unit and analyzes the collected exhaled mist,
A lung volume fraction measuring device configured to perform subject's lung volume fractionation and measure the subject's closing volume,
Breath mist analysis system characterized by being equipped with. The breath mist analysis system according to claim 3 , wherein the analyzer is a fluorescence emission spectrometer. A step of introducing the exhaled breath of the subject into an adsorbent and collecting the exhaled mist containing non-volatile and refractory substances released from blood or the like through the alveoli and the respiratory tract.
Steps to measure the exhaled volume of the subject,
A step of determining the suitability of the exhaled mist collection condition based on the measured exhaled volume each time is provided.
In the step of determining the suitability of the breath mist collection condition, the breath mist collection method is characterized in that the suitability of the breath mist collection condition is determined based on a threshold value set based on the closing capacity of the subject. .. The method for collecting exhaled mist according to claim 5 , wherein the adsorbent is an adsorbent capable of specifically capturing a target component having fluorescence. A breath mist analysis method according to claim 6 , wherein the breath mist collected by the adsorbent is analyzed by a fluorescence emission spectrometry method.

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