WO2012172011A1

WO2012172011A1 - Method for analysis of exhaled air

Info

Publication number: WO2012172011A1
Application number: PCT/EP2012/061342
Authority: WO
Inventors: Jens HOHLFELD; Heike BILLER; Hubert Lödding; Wilhelm Dunkhorst; Katharina Schwarz; Wolfgang Koch; Horst Windt
Original assignee: Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e. V.
Priority date: 2011-06-17
Filing date: 2012-06-14
Publication date: 2012-12-20
Also published as: DE102011077772A1; DE102011077772B4

Abstract

The present invention relates to a method for analysis of an exhalate, wherein the exhalate has particles with a minimum diameter d_min(exhalate) and a maximum diameter d_max(exhalate), said method comprising the measurement and/or determination of at least one physical variable of a defined first particle size fraction P1 of the exhalate, wherein the physical variable is chosen from the particle number concentration C1, the particle number flow F1, the particle number N1, the particle mass flow M1, the particle mass G1 or combinations thereof, wherein the first particle size fraction P1 is defined by a minimum diameter d_min(P1) and a maximum diameter d_max(P1), such that d_min(P1)>d_min(exhalate) and/or d_max(P1)<d_max(exhalate).

Description

Method for the analysis of exhaled air

The invention relates to a method for the analysis of exhaled air (Exhalat). Such a method may e.g. as a preparatory measure in the context of the early, non-invasive diagnosis of pathological changes of the peripheral areas of the lung (P AD, peripheral airway disease), as described e.g. in obstructive

Respiratory diseases (bronchial asthma, chronic obstructive

Pulmonary disease (COPD) or bronchiolitis obliterans or

Bronchiolitis obliterans syndrome in the context of organ rejection after

Lung transplantation occur, find application. Pathological changes of the lung periphery with bronchiolo alveolar

Fluid accumulation and / or inflammation occur i.a. also on at

Left ventricular failure, respiratory failure or iatrogenic lung injury from external ventilation (VILI, Ventilation induced lung injury). Furthermore, the method can be used as a preparatory measure in the context of the follow-up of lung diseases and for the efficacy testing and optimization of therapeutic methods for the treatment of

Lung diseases or the prevention of lung damage, especially in the context of long-term artificial respiration. Due to the steady increase in the prevalence of lung diseases

Early detection and follow-up have become an outstanding goal in pneumological diagnostics. Crucial here is the most specific and sensitive methodology that allows non-invasive information about the lung disease. The obvious analysis of exhaled air has become increasingly important in recent years. Ten years ago, after the fundamental measurability of non-volatile molecules (proteins, peptides) in human exhaled air could be demonstrated, there are now a number of descriptive findings on changes in the concentration of various

Inflammatory molecules in respiratory condensate in patients with lung diseases.

Nonvolatile molecules must necessarily have one

Drool formation process from the lung fluid out into the exhaled air. The properties of the exhaled aerosols are determined on the one hand by the properties of the droplets formed in the different lung compartments and on the other by the redeposition of already generated particles in the complex lung structures during the exhalation process. In normal breathing, the particle size spectrum usually has

Number maximum in the particle size range less than 1 μιη on.

One mechanism of drop formation in the lung, which is the dominant process in normal breathing, is the reopening of collapsed airway capillaries of the lung periphery and the associated abrupt separation of the lung

Epithelial fluid film lining the respiratory tract. In this liquid-liquid separation, droplets emerge, which can escape with the breath and can be detected. The process is shown in FIG.

Surface tension-related instabilities (Rayleigh plateau instabilities) of the lung fluid can lead to occlusion of small airways.

Liquid bridges or liquid plug 3, as shown in Fig. 1, make it a more energetically favorable state than the smooth lined with lung fluid film 2 capillary walls 1. This state is realized in particular during exhalation. During the inspiratory process, the transpulmonary pressure increases and it can be used to reopen the droplets 4 at

Exhalation process clogged airways come.

This drop formation process is of the processes that are e.g. in a forced

Exhalation or when speaking, coughing and sneezing become effective, too

differ. In the latter case, the droplets v. A. by tearing out of the surfactant film at high flow velocities and through the

Movement of the vocal cords. These processes lead to larger but significantly fewer particles.

The ventilated lung area where the airway can close is referred to as the "closing volume" (CV), when the lung volume at the end of an expiratory process (EELV, end expiratory volume) is less than the closing volume (EELV <CV) it for the realization of

Airway closures come. The particles or liquid droplets generated during the reopening are then carried outside with the respiratory flow during the next exhalation process. By varying the breathing depth is the

Likelihood of occlusion of the peripheral airways and thus the

Particle flow influenced. However, this is in direct relation to the individually different lung function parameters, in particular the

Vital capacity and the residual volume, which are shown in Fig. 2, which, as well as the closing volume for the healthy lung have a function of age.

FIG. 2 shows typical lung function parameters which, apart from the closure volume CV, are determined by conventional measuring methods such as spirometry or body plethysmography. These pulmonary function parameters shown in Fig. 2 are it is the lung capacity TLC ("total lung capacity"), the lung volume available for respiration VC ("vital capacity"), which is not ventilated

Residual volume RV (residual volume), tidal volume (VT) tidal volume, lung volume after respiratory distress after normal expiration FRC (functional residual capacity), closing volume CV, lung volume at End of the expiration process EELV ("end expiratory volume") and the exhaled volume EV ("expired volume").

There is a correlation between occlusion volume and changes in the peripheral airways. The measurement of the occlusion volume opens up the possibility of early diagnosis of diseases of the peripheral airways. Furthermore, it can be seen that in the ventilated patient, e.g. during anesthesia, a periodically recurring one enforced by the ventilation pattern

Airway obstruction may cause injury to the peripheral airway.

The occlusion volume can be measured by the administration and detection of noble gas boluses during pulmonary function examinations.

The events of the reopening of the respiratory tract can also be detected and quantified by the analysis of acoustic signals. They express themselves in so-called crackling noises (crackles). In principle, the analysis of "crackles" is suitable for the early detection of peripheral lung changes.

For the detection of changes in the peripheral airway dimensions in the context of lung diseases is the residence time-dependent

Sedimentation behavior of inhaled monodisperse particles in the

Used aerosol morphometry. In this method, the subject inhaled under a standardized breathing maneuver a monodisperse test aerosol with a particle diameter of 1 μιη, the concentration is determined as a function of the inspired volume. After a post-inspiratory respite During the subsequent exhalation, the decrease in concentration above the exhaled volume or lung depth is determined. The recovery rate of the particles as a function of the residence time of the particles allows the

Conclusion on the so-called mean effective airway diameter in the considered lung depth.

In addition to the morphometric examination of the lung, monodisperse aerosols also serve to analyze the convective transport of the respiratory gas as part of the so-called aerosol-bolus dispersion. Exhalation (dispersion) of an inhaled bolus of a low redeposition rate test aerosol allows conclusions to be drawn on pathological alterations of the respiratory tract associated with inhomogeneous ventilation of the lungs. Ventilation of the lungs is performed by the administration and detection of inert gas boli. Practically, both the gas measurements due to the complex detection and the acoustic measurements and the aerosol investigation are very complex.

It is an object of the present invention to provide a non-invasive method of analyzing respiratory air. Preferably, the method should be simple to perform and as a preparatory measure for subsequent lung diagnostics, e.g. may serve as an early diagnosis of pathological changes in the peripheral areas of the lung (PED).

This object is achieved by providing a method for analyzing an exhalate, wherein the exhalate particles having a minimum diameter d _m i _n (Exhalat) and a maximum diameter d _max (Exhalat) comprising the measurement or determination of at least one physical size a defined first particle size fraction PI of the exhalate, wherein the physical quantity is selected from the particle number concentration Cl, the

Particle number flow F 1, the number of particles N 1, the particle mass flow M 1, the Particle mass Gl or combinations thereof, wherein the first particle size fraction PI by a minimum diameter d _m i _n (Pl) and a maximum diameter dmax (PI) is defined such that d _m i _n (Pl)> d _m i _n (exhalation) and or

max (Pl) <d _max (exhalate).

The term "exhalate" refers to the total exhaled volume flow and / or the total exhaled volume and / or a partial volume flow and / or a partial volume of expired respiratory gas The term "particles" refers to the relative humidity in the environment of the aerosol either on liquid droplets or at a relative humidity well below 100% on solid particles present after the shrinkage of the droplets, consisting of the non-volatile substances of the liquid lining the respiratory tract as exhaled with exhalation. Due to their particle size distribution, these particles have a minimum diameter d _m i _n (exhalate) and a maximum diameter d _max (exhalate) which, of course, may vary depending on the subject or respiratory pattern or depth of respiration or activity Liquid droplet in the lungs has already been discussed above with reference to FIG. Typically, the maximum diameter d _max (exhalate) <5 μιη in normal breathing, coughing, sneezing, speaking or forced exhalation up to d _max (exhalate) <1mm.

In the context of the present invention, it has been found that a process in which a suitable particle size fraction (i.e.

Particle subset of all present in exhaled particles) defined and determined for this particle size fraction number concentration, number flow, mass flow and / or particle number, an efficient preparatory measure for subsequent lung diagnostics, eg an early diagnosis of pathological Changes in the peripheral areas of the lung (PED, peripheral airway disease) represents.

Preferably d _m i _n (Pl)> d _m i _n (exhalation) and d _max (Pl) <d _max (exhalation). Alternatively, it is also possible that d _mm (Pl)> d _mm (exhalate) and d _max (Pl) <d _max (exhalate)

Preferably dmin (Pl) and d _max (Pl) are chosen so that the first

Particle size fraction PI at most 95%, more preferably at most 90% of

Total number of particles in the exhalate includes. In a preferred

Embodiment d _m i _n (Pl) and d _max (Pl) are chosen so that the first

Particle size fraction PI 10% to 95%, more preferably 70% to 90% of

Total number of particles in the exhalate includes.

If the provision of the Exhalats by normal breathing or normal breathing in combination with at least one other different type, such as coughing, sneezing speaking or forced expiration, d _m i _n (Pl) is preferably μιη to a value in the range of 0.2 to 1 , 5 μιη, more preferably in the range of 0.3 μιη set to 0.6 μιη. If the exhalate is provided by coughing, sneezing or forced exhalation or a combination of at least two of these activities, dmin (P1) is preferably adjusted to a value in the range of 0.2 μm to 100 μm, more preferably in the range of 0.3 μm set to 10 μιη. The maximum particle diameter d _max (Pl) of the first particle size fraction can be selected, for example, such that it corresponds to d _max (exhalate), ie

max (Pl) = d _max (exhalate). As already stated above, the maximum diameter d _max (exhalate) is typically <5 μιη in normal respiration; for coughing, sneezing, speaking or forced exhalation, d _max (exhalate) <1 mm. The mass flow M 1 of the defined first particle size fraction PI can be the total particle mass flow or the particle partial mass flow of one or more specific components of the particles, such as certain proteins or salts. In the former case, the total mass of the particle is taken into account for the measurement or determination of the mass flow, while in the latter case only the mass of this specific component or components of the particle is taken into account for the measurement or determination of the mass flow.

The mass Gl of the defined first particle size fraction PI may be the total particle mass or the particle mass of one or more specific components of the particles, such as e.g. certain proteins or salts act. In the former case, the total mass of the particle is taken into account for the measurement or determination of the mass, while in the latter case only the mass of this specific component or components of the particle is taken into account for the measurement or determination of the mass.

Suitable devices for measuring or determining the

Particle number concentration, the particle number flow, the number of particles, the particle mass flow and the particle mass of aerosol particles are the

Specialist known. The measurement or determination can be made, for example, with a condensation nucleus counter or a scattered light counter or an impactor or a TEOM method or a quartz balance, a measuring device for particle size determination, preferably an optical particle counter

Duration spectrometer (aerodynamic particle classifier), an electric mobility spectrometer, a cascade impactor (mass determination via detection of electric current or quartz crystal balance) or a Combination of these devices, optionally in combination with a known to the expert analysis method for the determination of specific

Components of the particles, take place. Condensation core counters are known per se to those skilled in the art and are also commercially available. Usual condensation nucleus counters have one

Humidification zone in which there is a container of liquid, and a condensation zone in which the particle growth takes place by the vapor of the humidification zone condenses on the particle surface.

This measuring method with condensation nucleus counter permits, in contrast to pure stray light particle counts, the detection of particles with diameters smaller than 0.1 μm. Particles smaller than 0.1 μιη be because of their low

Scattered light signal with pure scattered light meters no longer detected. in the

Condensation core counter lets you pass through the submicron particles

Condensation of a supersaturated vapor (butanol, isopropanol, water) grow to sizes above a few microns. These particles can then be counted in a simple scattered light counter. From the count rate, RN, and the sampling volume flow, the concentration of the particles in the exhalate results. It is advantageous to perform the breathing maneuvers so that the tidal volume flow is always greater than the sampling volume flow. Then the number of particles emitted per breath results by simply adding up all

Counting events during the half-wave of exhalation and multiplication by the ratio of tidal volume and sample volume. The control of the measuring process, the recording of the resulting data and the necessary preferred are preferably carried out with a PC.

As mentioned above, condensation nucleus counters are commercially available. In many devices, however, the sampling volume flow at 0.11 rpm is so low that, given the overall low emission rates, the statistical reliability for the Recording the counting events can be negatively influenced. Furthermore, in the devices partly harmful vapors (such as butanol) are used, which then have to be filtered consuming. Moreover, commercial devices are expensive not least because of their high technical complexity for saturating the steam and subsequently controlled cooling. In a preferred embodiment, the condensation nucleus counter used has no humidification zone (ie no container with liquid). It is therefore preferred that the increase in the particle diameter in the condensation nucleus counter essentially, more preferably exclusively by condensation of the

Steam of the exhaled air takes place. This preferred embodiment thus utilizes the steam condensation of the already saturated with steam breathing air. Such a condensation nucleus counter is e.g. in EP 2 248 464 A1.

Preferably, the condensation nucleus counter is mounted in a device having a tube with inhalation and Exhalationszweig. Preferred is in

Inhalationszweig the tube, preferably in Exhalationszweig the tube, a filter, preferably attached to an absolute filter. The term "absolute filter" is used in the context of the present invention in its usual, the expert

used in common meaning and refers to a filter that deposits substantially all particles, with an efficiency better than 99.99%. With an absolute filter, it can be guaranteed that the inhaled air is particle-free and thus does not disturb the particle measurement on the exhalate.

Devices for measuring or determining the particle diameter or the particle size distribution of aerosol particles are also known in principle to the person skilled in the art.

In a preferred embodiment, an optical particle counter is used which counts the particles and classizes them. In addition to determining the size distribution of all exhaled particles in the measuring range of the meter allows such an optical particle counter and the selective detection of individual particle size fractions.

Optical particle counters count and classify particles according to their requirements

size-dependent scattered light intensity. For this purpose, the aerosol flow passes through a sample chamber, through which a powerful light beam is projected. The scattered light emitted by each individual particle is picked up by a receiving optical system and converted into a size class arrangement into a voltage pulse corresponding to the size of the detected particle. Such devices are generally known to the person skilled in the art.

According to further preferred embodiments, the measurement or

Determination of the particle number concentration Cl, the particle number flow Fl, the particle number Nl, the particle mass flow Ml or the particle mass G1 of the defined first particle size fractions PI of the exhalate also by a

Scattered light or a runtime spectrometer (aerodynamic particle classifier) or an electric mobility spectrometer done.

In a scattered light counter, e.g. a detection range over the intensity of the light beam of the transmitting optics and the selection of the detector or its

Detection threshold are set. This detection range can then be set so that it with the specified minimum diameter d _m i _n (Pl) and / or set maximum diameter d _max (Pl) of the first

Particle size fraction PI matches.

In a further preferred embodiment, the measurement or

Determination of the particle number concentration Cl, the particle number flow Fl, the particle number Nl, the particle mass flow Ml or the particle mass Gl of the defined first particle size fractions PI of the exhalate by a

Condensation core counter with a diameter d _m i _n (Pl) corresponding lower detection limit and / or a fixed maximum diameter dmax (Pl) upper detection limit.

In further preferred embodiments, the measurement or determination of the particle number concentration Cl, the particle number flow Fl, the particle number Nl, the particle mass flow Ml or the particle mass G1 of the defined first particle size fractions PI of the exhalate is carried out by a condensation nucleus counter or a scattered light counter or another of the abovementioned measuring methods a lower detection limit equal to or less than the fixed diameter d _m i _n (Pl), or an upper detection limit equal to or greater than the specified maximum diameter d _max (Pl), the minimum diameter d _m i _n (Pl) and / or the maximum diameter dmax (Pl) defined first particle size fraction PI of the exhalate by a suitable pre-separation or fractionation of the exhaled aerosol

provided.

A typical measuring example is shown in FIG. FIG. 3 shows the time profile of the number flow F1 for differently defined particle size fractions PI. In addition, Figure 3 shows the total number of flow of all the exhaled particles, and in the upper half the respiratory flow, the negative half wave indicating exhalation (i.e., exhalation). The simultaneous measurement of the particle concentration or the particle number flow and the

Particle size distribution, i. the particle concentration or the

Particle number flow in individual size classes makes it possible to determine the number flow and the size distribution of the aerosols in the exhalation branch of the respiration.

In a preferred embodiment, the method further comprises:

the measurement or determination of a physical quantity for all particles of the exhalate or for at least one defined second particle size fraction P2, which is defined by a minimum diameter d _m i _n (P2) and a maximum diameter d _max (P2), where

and or

wherein the physical quantity is selected from the particle number concentration C2, the particle number flow F2, the

Particle number N2, the particle mass flow M2, the particle mass G2 or combinations thereof; and

the determination of at least one parameter value from a

mathematical relationship comprising at least one of the physical quantities C1, F1, N1, M1 or Gl and at least one of the physical quantities C2, F2, N2, M2 or G2.

The physical variable which is determined for all exhalate particles or the defined second particle size fraction P2 is preferably the same physical quantity which is also determined for the defined first particle size fraction PI.

As already noted above with regard to the particle size fraction PI, the mass flow M2 may be the total particle mass flow or the mass flow M2

Particle mass flow of a specific component of the particles, e.g.

certain proteins or salts act. In the former case, the total mass of a particle is taken into account for the measurement or determination of the mass flow, while in the latter case only the mass of this specific component of the particle is taken into account for the measurement or determination of the mass flow.

As already mentioned above with regard to the particle size fraction PI, the mass M2 may be the total particle mass or the particle partial mass of a specific component of the particles, such as certain proteins or salts. In the former case, the total mass of a particle is taken into account, while in the latter case only the mass of this specific component of the particle is taken into account. With regard to suitable devices and conditions for the measurement or

Determination of C2, F2, M2, N2 and / or G2 can be referred to the above comments on the first particle size fraction PI.

Therefore, the measurement or determination of the particle number concentration C2, the particle number flow F2, the particle number N2, the particle mass flow M2 or the particle mass G2, for example, with a Kondensationskernzähler or a scattered light counter or an impactor or a TEOM method or a quartz balance, a particle size measuring device, preferably an optical particle counter, a runtime spectrometer (aerodynamic particle classifier), an electric mobility spectrometer, a

Cascade impactor (mass determination via detection of electric current or quartz crystal balance) or a combination of these devices,

optionally in combination with a person skilled in the art

Analytical methods for the determination of specific components of the particles, take place.

According to a further preferred embodiment, the measurement or

Determination of the above-mentioned physical quantities also by a

Condensation core counter, which detects the particles via a scattered light counter, in combination with another scattered light counter, the first

Scattered light counter is mounted in a region in which no growth in size of the particles has yet taken place by condensation of a vapor (e.g.

immediately before the condensation nucleus counter or at the very beginning of the

Kondensationskernzählers) and the second the Kondensationskernzähler associated scattered light counter is mounted as usual at the end or after the condensation zone. For example, if the scattered light meters have a lower detection limit of 0.5 μιη, the first scattered light counter of the total number of exhaled particles only detects those with a diameter of at least 0.5 μιη, during the second scattered light counter and the fine particles below 0.5 μηι detected because at the end or after the condensation zone and the very small particles have grown to a diameter of more than 0.5 μιη and thus are detectable in the scattered light counter.

According to a further preferred embodiment, the measurement or

Determination of the above physical quantities also by a

Combined two scattered light counter with different lower and / or upper detection limit.

As already noted above, the determination of the detection range of a scattered light counter can be effected via the intensity of the light beam of the transmitting optics and the selection of the detector or its detection threshold. According to further preferred embodiments, the measurement or

Determining the abovementioned physical quantities also by any other combination of two identical or different devices, the devices already mentioned above for the measurement or determination of the first particle size fraction PI.

The characteristic value can be determined by establishing a mathematical relationship comprising at least one of the physical quantities C1, F1, N1, G1 or M1 and at least one of the physical quantities C2, F2, N2, G2 or M2. The values determined or measured for Cl, Fl, Nl, Gl or Ml and the values determined or measured for C2, F2, N2, G2 or M2 are then inserted into this mathematical relationship and a characteristic value is obtained.

For example, this mathematical relationship may be a

Quotienten from Cl and C2 or Fl and F2 or Nl and N2 or Ml and M2 or Gl and G2 act. Other mathematical relations for the determination of the

Characteristic value are also possible in the context of the present invention (Addition, subtraction or multiplication of the physical quantities or corresponding combinations thereof).

Such representations can be found in FIGS. 4a-4d and 5.

FIGS. 4a-4d show the ascertained characteristic values as a function of the already exhaled volume in the exhalation process. The characteristic values were determined from the ratio of the number flow of the particles with a minimum diameter dmin (P1) of 0.5 μm to the total number flow of all exhaled particles. Furthermore, the figures still show the measured total number of currents.

FIG. 4a (top left) shows the progression of the characteristic values as a function of the exhaled volume for a healthy non-smoker, FIG. 4b (top right) for a healthy smoker, FIG. 4c (bottom left) for a subject with mild COPD (stage I), FIG 4d (bottom right) for a subject with severe COPD (stage III).

FIG. 5 shows the characteristic values, which were determined as the quotient of the total number of particles to the number of particles> 0.5 .mu.m.sup.-1 the entire exhalation process for different subjects, as a function of the vital capacity of the subjects.

It can be seen that the course of the particle size ratio as a quotient of the number of particles> 0.5 μιη to the total number of particles in the time interval considered as a function of the already exhaled volume in

Exhalation process for healthy smokers and people with increasing

Severity of COPD changed significantly, especially towards the end of

Exhalation. A simple example of the mathematical evaluation is the formation of the quotient of the total number of particles to the number of particles> 0.5 μιη over the entire Exhalationsvorgang. But that would be just as possible Forming a difference in particle size ratio at a given time or expired volume at the beginning of the exhalation process and at the end of the exhalation process. In Figure 5, the characteristic values show differences for healthy nonsmokers and patients with chronic obstructive pulmonary disease and can therefore be used in principle as a biomarker for changes in the lung.

Figure 6 shows the exhaled particle concentration and simultaneously the

Particle size ratio in the considered time interval as a function of the already exhaled volume in the exhalation process for a subject with moderate COPD.

In artificial respiration, the properties of the endogenously generated aerosols in exhalation of the patient or changes in the same statements about the ventilation state of the lungs and the occurrence of recurrent

Airway closures, and thus be used to optimize the ventilation strategy to avoid ventilator-induced lung injury. In the context of the process according to the invention, the exhalate can be replaced by a

Breath or alternatively be provided by several breaths. If the exhalation is provided by several breaths, it may be preferred that at least two different breathing patterns (different breathing depths or respiratory flow rates) are used.

The method can be carried out for one or more breathing patterns with one or more breaths, optionally with simultaneous measurement or determination of at least one respiratory physiological variable and / or at least one lung function parameter. The measurement or determination of the abovementioned physical variables during a breath is preferably carried out several times at defined time intervals.

Optionally, the method according to the invention can also be the measurement or determination of at least one lung function parameter and / or at least one

physiological size, and the determination and / or graphical representation of the characteristic value or the characteristic values as a function of

respiratory physiological size or size or the pulmonary function parameter (s).

The measurement or determination of at least one pulmonary function parameter or a respiratory physiological parameter may e.g. by spirometry and / or

Bodyplethysmography done. Preferably, a pneumotachograph or acoustic volume flow meter, e.g. an ultrasound spirometer, used. These measuring methods are known in principle to the person skilled in the art. For example, the lung capacity TLC available for respiration may be used as appropriate lung function parameters or respiratory physiological parameters

Lung volume VC, the non-ventilatable residual volume RV, the

Tidal tidal volume VT, lung volume after respiratory distress after normal expiration FRC, lung volume at the end of expiration EELV, exhaled volume or respiratory flow rate.

In FIGS. 4 and 6, the representation of the characteristic values was carried out by way of example as a function of the exhaled volume and in FIG. 5 as a function of the vital capacity.

In a preferred embodiment, the method according to the invention can also be used for the analysis of exhalate aerosol particles, preferably non-volatile substances, such as biological molecules (eg proteins, DNA / RNA, viruses, bacteria) or non-biological substances (eg metals, pharmaceutical agents). Find application. For conclusions on the concentrations or Determination of the concentration of these non-volatile substances in the fluid lining the respiratory tract and the comparison of results

different people is the knowledge of the number flow and the

Particle size distribution or the emitted aerosol mass flow required. This differs considerably from person to person. It has been found that for a sufficiently precise concentration determination of the substances in the aerosol particles or the normalization of the

Analytical results is sufficient to determine the above physical size such as the particle number flow Fl only for a suitably defined particle size fraction PI. Minimum diameter d _m i _n (Pl) and maximum diameter dmax (Pl) of this particle size fraction are thereby preferably defined so that the fraction of the larger particles is detected, which contains the majority of the emitted aerosol mass flow. For example, d _m i _n (Pl) can be set to a value in the range of 0.2 μιη to 0.6 μιη and the maximum

The particle diameter d _max (Pl) can be chosen, for example, to be d _max (exhalate), ie d _max (Pl) = d _max (exhalate).

The results of such an application of the method according to the invention are shown in FIGS. 7a and 7b.

In FIG. 7a, the deposition of the fraction of particles> 0.2 μm from the exhaled aerosol for different collection periods (1 to 5 minutes) was carried out for two subjects by means of a slot impactor on a membrane under simultaneous conditions

Determination of the number of particles (top row: test person 1, middle row: test person 1 repetition; lowest row: test person 2). As the number of deposited particles increases, a clear increase in the detection intensity can be seen, and a good reproducibility of the results for subjects 1 in the case of

Repeat the measurement. FIG. 7b shows the optical density, determined by optical evaluation, of the individual samples as a function of the associated deposited number of particles. There is a clear increase in the optical density as the number of particles increases.

Preference is given to measuring or measuring in the method according to the invention

certain values of the particle number concentration Cl, the particle number flow Fl, the particle mass flow Ml, the particle mass Gl or the particle number Nl and optionally the particle number concentration C2, the particle number flow F2, the particle mass flow M2, the particle mass or the particle number N2 stored in a data memory and / or a computing unit for the determination of

Characteristic values supplied.

The process according to the invention is preferably a

Computer-aided method using a computing unit, which is program-technically arranged so that on the basis of the measured or

determined values of the particle number concentration Cl, the particle number flow Fl, the particle mass flow Ml, the particle mass Gl and / or the particle number Nl and optionally the particle number concentration C2, the particle number flow F2, the particle mass flow M2, the particle mass G2 and / or the particle number N2 the determination of at least a characteristic value can be made.

According to a further aspect, the present invention preferably provides a computer program for carrying out the method described above and a data carrier with this computer program.

Claims

claims

A method of analyzing an exhalate, wherein the exhalate has particles with a minimum diameter d _m i _n (exhalate) and a maximum

Diameter d _max (exhalate) comprising the measurement or

Determining at least one physical quantity of a defined first particle size fraction PI of the exhalate, the physical quantity being selected from the particle number concentration Cl, the

Particle number flow Fl, the particle number Nl, the particle mass flow Ml, the particle mass G1 or combinations thereof, wherein the first

Particle size fractions PI by a minimum diameter d _m i _n (Pl) and a maximum diameter d _max (Pl) is defined so that d _m in (Pl)> d _min (Exhalat) and / or d _max (Pl) <d _max (exhalation).

The method of claim 1, wherein d _m i _n (Pl)> d _m i _n (exhalate) and d _max (P i) <d _max (exhalate); or where dmi "(Pl)> d _min (exhalate) and d _max (Pl) <d _max (exhalate).

The method of claim 1 or 2, wherein d _m i _n (Pl) and d _max (Pl) are selected so that the defined first particle size fraction PI comprises a maximum of 95% of the total number of particles in Exhalat.

The method according to any one of the preceding claims, further comprising the measurement or determination of a physical quantity for all particles of the exhalate or for at least one defined second

Particle size fraction P2, which is defined by a minimum diameter d _m i _n (P2) and a maximum diameter d _max (P2), wherein

and / or _dmax (P2) 7i: _dmax (Pl), wherein the physical quantity is selected from the particle number concentration C2, the

Particle number flow F2, the particle number N2, the particle mass flow M2, the particle mass G2 or combinations thereof; and the determination of at least one characteristic value from a mathematical relationship comprising at least one of the physical quantities C1, F1, N1, G1 or M1 and at least one of the physical quantities C2, F2, N2, G2 or M2.

The method according to any of the preceding claims, wherein the measurement or determination of the physical quantity by a

Condensation core counter, a scattered light counter, an impactor, a TEOM method, a quartz balance, a measuring device for

Particle size determination, preferably an optical particle counter, a runtime spectrometer (aerodynamic particle classifier), a

electric mobility spectrometer, a cascade impactor, or a combination of these devices.

The method of any one of the preceding claims, wherein the exhalate is provided by a breath; or alternatively provided by several breaths, preferably at least two

different breathing patterns are used.

The method according to one of the preceding claims, wherein the measurement or determination of the particle number concentration Cl, of the

Particle number flow Fl, the particle number Nl, the particle mass flow Ml or the particle mass G1 and optionally the measurement or determination of the particle number concentration C2, the particle number flow F2, the particle number N2, particle mass flow M2 or the particle mass G2 during a breath several times at defined time intervals.

The method of any one of the preceding claims, further comprising measuring or determining at least one lung function parameter and / or a respiratory physiological size; and optionally the determination and / or graphical representation of the characteristic value or the

Characteristic values as a function of the respiratory physiological size (s) or the lung function parameter or the lung function parameters.

9. The method according to claim 4, wherein the method is carried out using a computing unit that is set up in the program in such a way that on the basis of the measured or determined values of the particle number concentration Cl, the particle number flow F 1, of the

Particle mass flow Ml, the particle mass Gl and / or the particle number Nl and the measured or determined values of

Particle number concentration C2, the particle number flow F2, the

Particle mass flow M2, the particle mass G2 and / or the particle number N2, the determination of the at least one characteristic value can be carried out.

10. A computer program for carrying out the method according to claim 9.