WO2016159863A1 - Method and device for analyte sampling and analyte concentration determination from exhaled breath - Google Patents

Method and device for analyte sampling and analyte concentration determination from exhaled breath Download PDF

Info

Publication number
WO2016159863A1
WO2016159863A1 PCT/SE2016/050258 SE2016050258W WO2016159863A1 WO 2016159863 A1 WO2016159863 A1 WO 2016159863A1 SE 2016050258 W SE2016050258 W SE 2016050258W WO 2016159863 A1 WO2016159863 A1 WO 2016159863A1
Authority
WO
WIPO (PCT)
Prior art keywords
capillary
analyte
adsorbent surface
exhaled breath
sampling device
Prior art date
Application number
PCT/SE2016/050258
Other languages
French (fr)
Inventor
Mohamed Abdel-Rehim
Original Assignee
Mohamed Abdel-Rehim
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Mohamed Abdel-Rehim filed Critical Mohamed Abdel-Rehim
Publication of WO2016159863A1 publication Critical patent/WO2016159863A1/en

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/497—Physical analysis of biological material of gaseous biological material, e.g. breath
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/08—Detecting, measuring or recording devices for evaluating the respiratory organs
    • A61B5/082—Evaluation by breath analysis, e.g. determination of the chemical composition of exhaled breath
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/08—Detecting, measuring or recording devices for evaluating the respiratory organs
    • A61B5/097—Devices for facilitating collection of breath or for directing breath into or through measuring devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/22—Devices for withdrawing samples in the gaseous state
    • G01N2001/2244—Exhaled gas, e.g. alcohol detecting
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/22—Devices for withdrawing samples in the gaseous state
    • G01N1/2273—Atmospheric sampling
    • G01N2001/2276—Personal monitors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • G01N2001/2826—Collecting by adsorption or absorption
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/0004—Gaseous mixtures, e.g. polluted air
    • G01N33/0009—General constructional details of gas analysers, e.g. portable test equipment

Definitions

  • the present invention relates to the field of analyte sampling from exhaled breath
  • PCT/US2012/027778 discloses methods and devices for collecting and measuring volatile organic compounds from breath. Briefly, the apparatus contains a solid-phase
  • SPME microextraction
  • Trefz et al. (Anal Bioanal Chem 2013 vol 405, pages 3105-31 15) describe evaluation of needle trap microextraction (NTME) in breath analysis. While the results were
  • NTME neuropeptide
  • An object of the present invention is to provide improved and/or alternative methods and devices for sample collection and analysis from breath samples.
  • phenyl methyl silicone refers to those silicon compounds in which two organic groups (phenyl + methyl) and oxygen are combined with silicon [-0-SiR 2 R 2 -] n , wherein each R 2 is independently selected from methyl and phenyl.
  • An exemplary preferred phenyl methyl silicone (dimethyl diphenyl siloxane) is shown below as Formula (I)
  • polyethylene glycol refers to a chemical compound composed of repeating ethylene glycol units H-[0-C 2 H 2 -j n OH.
  • a sol-gel of silica refers to the condensation of tetraethyl orthosilicate (TEOS) after hydrolysis in water [Si(OC2H5) 4 + H 2 0 ⁇ HO-Si(OC2H5) 3 + C 2 H 5 -R-OH].
  • TEOS tetraethyl orthosilicate
  • Figure 1 depicts a schematic drawing of a sampling device of the invention.
  • Glass or metal capillary length e.g. 50-100 mm.
  • Polymeric coating (3) Inside diameter e.g. 3-5 mm.
  • Figure 2 depicts a schematic flowchart of a sampling method of the invention. (5) Flow (breath) inlet. (6) Flow outlet.
  • FIG. 3 Schematic representation of desorption of the volatile compounds by heated injector into GC-MS. (7) Coated capillary liner in the injector body. (8) Heated gas chromatography injector. (9) Gas chromatography oven. (10) Mass spectrometer.
  • Figure 4 depicts exemplary chromatograms (top) and mass-spectra (bottom) of volatile compounds captured from exhaled breath by the method and device of the invention.
  • Figure 5 depicts the structure of an exemplary sol-gel to be coated in capillary.
  • the present invention relates to a method and a device for sampling analytes, such as volatile compounds from exhaled air, in particular biomarkers or compounds of toxicological or forensic interest.
  • the analytes which are typically volatile compounds are initially captured on sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof. Subsequently, the analytes may be released from the surface for analysis with any known suitable method, such as GC-MS or the like.
  • the present invention relates to the following aspects.
  • a method for collecting and determining the amount of an analyte sample from exhaled breath comprising the steps of:
  • a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof;
  • the determination step comprises the step (c ' ) of desorbing the bound analyte from the adsorbent surface by means of heat and/or a solvent.
  • the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of heat, preferably at 200-300°C.
  • the determination step comprises the step (c') of desorbing the bound anaiyte from the adsorbent surface by means of heating from 50-250X.
  • the determination step comprises the step (c') of desorbing the bound anaiyte from the adsorbent surface by means of a solvent such as acetonitrile and/or methanol.
  • the method according to aspect 1 wherein the contacting step is conducted during a time span of no more than 1 minute, preferably during less than 50 seconds, 45 seconds, 30 seconds, 20 seconds or 10 seconds.
  • the adsorbent surface comprises two compounds selected from the groups consisting of a phenyl methyl silicone, polyethylene glycol and a sol-gel of silica, coated on a first and a second part of the adsorbent surface of the capillary .
  • the present invention further relates to the following items:
  • a method for collecting an anaiyte sample from exhaled breath comprising the steps of:
  • a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof;
  • anaiyte is a marker of toxicological interest.
  • the analyte is a marker for a substance of abuse, such as an opiate, a cannabinoid, an
  • amphetamine or ***e, or a marker of exposure to environmental or
  • the adsorbent surface comprises a compound selected from the groups consisting of a phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.
  • analyte to be sampled is selected from the group consisting of pentane, decane, acetaldehyde and octane.
  • the method is further for determining the amount of an analyte in a sample from exhaled breath, and comprises the step (c) of determining the amount of bound analyte.
  • the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface.
  • the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of heat, preferably at 200-300°C or a solvent such as acetonitrile or methanol.
  • the determination step comprises the step (c") of determining the analyte by means of GC, LC, MS or a combination of these methods
  • the determination step comprises the step (c") of determining the analyte by means of GC-MS or LC-MS.
  • sampling device is a device according to any one of items 15-31.
  • the contacting step involves transporting the exhaled breath through the capillary by the force of exhalation of the subject from which the breath is exhaled.
  • a sampling device for collecting an analyte sample from exhaled breath comprising a capillary with an adsorbent surface coated on the inner surface thereof. 16.
  • the adsorbent surface comprises an adsorbent selected from the group consisting of phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.
  • the adsorbent comprises phenyl methyl silicone.
  • the capillary wall comprises a glass, a metal or a plastic, preferably glass.
  • the adsorbent surface is 200-300 ⁇ in thickness.
  • the device comprises a mouthpiece and/or a handle, preferably a mouthpiece.
  • a method for diagnosis of a disease or condition such as cancer, in particular lung cancer comprising detecting a biomarker by a method involving a method according to any of items 1-13.
  • a method for toxicological screening comprising detecting an analyte being a marker for chemical exposure by a method involving a method according to any of items 1-13.
  • the present invention relates to a method for collecting an analyte sample from exhaled breath of a subject, comprising the steps of:
  • a sampling device comprising an adsorbent surface
  • the sampling device comprises a capillary with an adsorbent surface coated on the inner surface thereof, to which the analyte binds.
  • the capillary may have any of the properties such as dimensions or materials discussed below under "Properties of the capillary" of the device of the second aspect of the invention.
  • the adsorbent surface may comprise a compound selected from: a phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.
  • the adsorbent surface comprises phenyl methyl silicone or polyethylene glycol, as these adsorbents are capable of capturing both hydrophobic and hydrophilic analytes. Both are stable at high temperature (>250°C) and are capable of capturing analytes of interest with different polarity.
  • the analyte to be sampled may be any analyte present in exhaled breath, capable of binding to the particular adsorbent present in the sampling device.
  • the analyte to be sampled may be a volatile compound.
  • the analyte may be a biomarker, such as a biomarker for a disease or a condition.
  • the analyte is a biomarker for cancer, such as lung cancer, or a disease of the respiratory system, such as astma or chronic obstructive lung disease including prodromal stage thereof.
  • the analyte may be a compound of interest for toxicological analysis.
  • the analyte may be a marker for substance abuse or drug overdose, a marker for exposure to environmental or occupational hazardous substances, a marker for exposure to a chemical weapon, or the like.
  • the analyte is selected from pentane, decane, acetaldehyde and octane.
  • the method may further comprise the step (c) of determining the amount of bound analyte.
  • the determination step may comprise the step (c') of desorbing the bound anaiyte from the adsorbent surface.
  • the desorption step (c') may preferably be performed by means of heating the sampling device, preferably at 200-300°C.
  • the desorption step (c') may preferably be performed by means of heating the sampling device, at 50-250°C, such as at 50-200X, or at 100 -250°C.
  • the desorption step (c') is performed by means of heating the sampling device at 100 -250°C.
  • a distinct advantage of the capillary format (see further discussion below under the second aspect) of the sampling device used in the method is that desorption can be conveniently achieved in a standard GC instrument by inserting the capillary into the injector chamber.
  • a solvent such as acetonitrile or methanol can be used for desorption.
  • the solvent containing the desorbed analyte can then proceed to analysis.
  • the determination step comprises the step (c") of determining the analyte by means of gas chromatography (GC), liquid chromatography (LC), mass spectrometry (MS) or a combination of these methods.
  • GC-MS or LC-MS is preferred, GC-MS being the most preferred since the capillary can be inserted in the injector for convenient analyte desorption by heating, as discussed above.
  • the sampling device may be a sampling device according to the second aspect of the present invention described below.
  • the contacting step involves transporting the exhaled breath through the capillary solely, substantially solely, substantially or mainly by the force of the subject's exhalation, without the use of collection reservoirs such as bags or pumps of any kind.
  • the contacting step is performed for no longer than 2 minutes by the exhaling patient in a time span of no more than approximately 1-2 minutes, such as in a time span of between 1 second-30 seconds, 30 seconds- 1 minute, 1 minute -1.5 minutes, or 1.5 minutes - 2 minutes. I.e. the patient exhales for no longer than approximately 2 minutes, such as in a time span of between 1 second-30 seconds, such as for no longer than 10 seconds, 20 seconds, 30 seconds, 1 minute , 1.5 minutes, or 2 minutes.
  • the contacting step can take as little as 30 seconds by the exhaling patient in a time span of no more than approximately 10 - 30 seconds, such as in a time span of between 1-10 seconds, 10 seconds-20 seconds, 20 seconds- 30 seconds.
  • the patient exhales for less than approximately 10 seconds.
  • the present invention relates to a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof, characterized in that the adsorbent surface comprises an adsorbent selected from the group consisting of phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.
  • the adsorbent surface comprises phenyl methyl silicone or polyethylene glycol, as these adsorbents are capable of capturing both hydrophobic and hydrophilic anaiytes. Both are stable at high temperature (>250°C) and are capable of capturing anaiytes with different polarity.
  • the device may optionally comprise a mouthpiece and/or a holder to facilitate handling.
  • the present invention relates to a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof, characterised in that the adsorbent surface comprises a first polymer immobilized on the first part of the inner surface and a second polymer immobilized on the second part of the inner surface.
  • the capillary may have an inner cross-sectional dimension of 2-10 mm, preferably 2.5-8 mm, more preferably 2.5-6 mm, most preferably 3-5 mm.
  • the cross-section may have any shape compatible with the application, such as circular, oval, triangular, square, rectangular, pentagonal and octagonal or the like, in most applications however, a substantially circular cross-section is preferable from a practical point of view in particular in terms of handling and availability.
  • the same considerations apply to the outer dimensions of the capillary. It is preferred the outer cross-section is substantially circular to facilitate interfacing with common existing instruments such as GC. However, it is contemplated that in some instances the capillary could be integrated in a larger block or a module, which could be adapted for interfacing with a particular instrument. In such case, the outer dimensions could vary greatly.
  • the thickness of the capillary wall may be 0.001-2 mm, 0.003-0.5 mm, or 0.005-0.1 mm.
  • the capillary wall thickness is most relevant if easy interface with existing instruments is desired. It is however understood that in cases where the capillary is integrated in a block or a module, there is substantially more freedom in choosing the wall thickness.
  • the capillary, or the channel therein may be 4-10 cm in length, preferably 4-8 cm, more preferably 6-9 cm, most preferably 7-10 cm.
  • the preferred measures provide a
  • the capillary is preferably straight, as this provides advantages in terms of storage (In a box), general handling, sampling (lowest possible back pressure) and in particular interface with standard analytical instruments such a GC.
  • Most instruments adapted for samples in capillary are designed on the premise that capillaries are straight, having essentially circular cross-section and having a certain external diameter (about 0.5-1.5 mm) and having a certain length (about 5 cm). It is advantageous to adhere to designs adapted to suit the instruments intended to be used for downstream analysis.
  • the capillary may also be curved, having one or more bends, or be of a spiral shape. This applies in particular in cases discussed above where the capillary is integrated in a block or a module.
  • the device is arranged such that the exhaled breath can be transported through the device by the force of the subject's exhalation, eliminating the need for collection reservoirs such as bags or pumps of any kind.
  • the capillary may not be too narrow, too winded or too long, since otherwise the pressure needed to transport a sufficient volume of exhaled breath during the short time that the subject is able to exhale becomes too large.
  • too short or too wide capillaries do not provide unsufficient adsorption capacity.
  • the structural capillary wall may be made of heat-tolerant inert material such a glass, a ceramic or a metal, allowing for both heat-based and solvent-based desorption.
  • inert material such as glass, a ceramic or a metal
  • PEEK poly(ether etherketone)
  • PTFE poly(tetrafluoroethylene)
  • the adsorbent surface may be 100-4QGpm in thickness, more preferably 200-300 ⁇ .
  • the capillary to be coated may be pre-treated, for instance a glass capillary surface may be activated by acid/base treatment.
  • the polymer to be coated is dissolved in a volatile solvent, such as acetone or dichloromethane.
  • Capillary to be coated may then be filled with the polymer solution and left until the solvent has evaporated.
  • the capillary may then be subjected to a heat treatment.
  • the present invention provides the advantages of a) ease of use, b) low cost, c) fast (possible to get results at the same day); d) potential for being used as screening method, e.g. for early stage lung cancer.
  • Example 1 Manufacture of devices for analyte sample collection
  • capillary inside surface of a glass capillary was activated by acidic and basic solution before the immobilization of the liquid polymer (phenyl methyl silicone or poly ethylene glycol).
  • the capillary was filled with strong acid such as hydrochloric acid [0.1 M] and left for 60 minutes at room temperature. After 60 min the capillary was rinsed with Milli-Q water and was left to dry. After drying, the capillary was filled with sodium hydroxide [1 M] for 60 minutes to activate the siianol groups (hydroxy! groups, OH). After an additional rinse with Milli-Q water, the capillary was left to dry.
  • the polymer [phenyl methyl silicone (OV 17 from Sigma-Aldrich) or PEG 400 (Sigma- Aldrich)] to being used for coating was dissolved in acetone or dichloromethane to a concentration of about 2.0 mg/mL to facilitate the coating procedure.
  • the activated capillary was filled with the polymer solution and was left for some hours at room temperature for evaporating of the acetone or dichloromethane. The capillary was then left over night at 40 degrees. The next day the capillary was heated-treated by heating from 40 degrees up to 250 degrees and left for 2 hours at 250 degrees.
  • TEOS tetraethyl orthosilicate
  • APTES 3-aminopropyl)triethoxysilane
  • the capillary was activated as mentioned above and then was filled with sol-gel solution and left at room temperature for 24 hours. After that the capillary was incubated at 55°C for 36 hours.
  • Breath collection All patients and controls were instructed to not eat anything 4 hours before the breath test. Each study subject performed tidal breathing of unfiltered room air for some minutes. During this time, they inhaled through their nose and exhaled through their mouth into the coated capillary obtained according to Example 1. There were no flowers or plants in the rooms, so the VOCs (volatile compounds) in the breath were considered as originating from the subjects themselves. The breath samples were stored in room temperature.
  • Example 2 The capillary from Example 2 with collected volatile compound was inserted into a GC injector chamber.
  • the injector was programmed heated from 50°C to 250°C.
  • the adsorbed analytes were thermally desorbed into a GC column separation and then detected identified by mass spectrometry. Results for detection of certain volatile compounds is shown in Figure 4A-D.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Pathology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Pulmonology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Food Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Urology & Nephrology (AREA)
  • Physiology (AREA)
  • Medicinal Chemistry (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Medical Informatics (AREA)
  • Surgery (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

A method for collecting an analyte, such a volatile compound or biomarker sample from exhaled breath, comprising the steps of providing a sampling device comprising an adsorbent surface; and contacting exhaled breath from the subject to be sampled with the adsorbent surface whereby the analyte to be sampled binds to the adsorbent surface; characterized in that the sampling device comprises a capillary with the adsorbent surface coated on the inner surface thereof. A sampling device for collecting an analyte sample from exhaled breath comprising a capillary with an adsorbent surface coated on the inner surface thereof, characterized in that the adsorbent surface comprises an adsorbent selected from phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.

Description

METHOD AND DEVICE FOR ANALYTE SAMPLING AND ANALYTE CONCENTRATION DETERMINATION
FROM EXHALED BREATH
TECHNICAL FIELD
The present invention relates to the field of analyte sampling from exhaled breath,
BACKGROUND TO THE INVENTION
The early diagnosis of lung cancer is a critical issue to improve survival. Today different methods for diagnosis are used in clinical practice, including blood tests, X-ray, computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET). In the last two decades, a huge research interest has been focused on biomarker discovery for different cancer diseases. Exhaled breath is a potential source to find new biomarkers for lung cancer. In addition the non-invasive and safe nature of exhaled breath analysis allows for repeated samples to be taken within short time interval without discomfort for the patients. Thus there is an enormous interest to find new methods of breath analysis for detecting biomarkers. It is also of interest to capture samples for toxicological analysis from exhaled breath, in particular markers for exposure to substances of abuse, to environmental or occupational hazardous chemicals, and to chemical warfare agents.
It is known that a number of pathological conditions (such as asthma and lung cancer) can be diagnosed by analysing various analytes, such as volatile compounds present in exhaled breath from the patient to be diagnosed. In practice however, collecting such volatile biomarker samples from exhaled breath is not trivial. The analytesin breath are present at trace levels, which in combination with high humidity present makes the sampling, storage and transport of samples challenging, and as discussed in the recent review by Lourenco and Turner (Metabolites, 2014, 4, 465-498) major issues still remain (see Chapter 4 in Lourenco and Turner ).
PCT/US2012/027778 discloses methods and devices for collecting and measuring volatile organic compounds from breath. Briefly, the apparatus contains a solid-phase
microextraction (SPME) fibre in a housing through which the patient to be sampled exhales. However, the binding capacity of SPME fibres is limited and the binding is slow making it difficult to collect samples directly from exhaled breath without need for an intermediate store such as a bag or a bottle. Another major disadvantage is low sensitivity of the SP E method, again necessitating large volumes of sample. Additionally, in many cases the instability of the fibre is significant issue.
Trefz et al. (Anal Bioanal Chem 2013 vol 405, pages 3105-31 15) describe evaluation of needle trap microextraction (NTME) in breath analysis. While the results were
encouraging, it was concluded that several factors strongly affected the analytical results, and that all materials, as well as sampling and calibration procedures have to be defined before NTME can be applied in a practical setting. NTME requires fairly high pressure for sample application making it inpractical for collecting samples directly from exhaled breath.
Therefore, there is still need in the art for robust methods and device for sample collection and analysis from breath samples of analytes such as volatile compounds or biomarkers. An object of the present invention is to provide improved and/or alternative methods and devices for sample collection and analysis from breath samples. DEFINITIONS
The term phenyl methyl silicone refers to those silicon compounds in which two organic groups (phenyl + methyl) and oxygen are combined with silicon [-0-SiR2R2-]n, wherein each R2 is independently selected from methyl and phenyl. An exemplary preferred phenyl methyl silicone (dimethyl diphenyl siloxane) is shown below as Formula (I)
Figure imgf000003_0001
Formula (I)
The term polyethylene glycol refers to a chemical compound composed of repeating ethylene glycol units H-[0-C2H2-jnOH.
The term a sol-gel of silica refers to the condensation of tetraethyl orthosilicate (TEOS) after hydrolysis in water [Si(OC2H5)4 + H20→ HO-Si(OC2H5)3 + C2H5-R-OH].
in the present context the term contacting is used interchangeably with the term sampling.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 depicts a schematic drawing of a sampling device of the invention. (1) Glass or metal capillary (length e.g. 50-100 mm). (2) Polymeric coating (3) Inside diameter e.g. 3-5 mm. Figure 2 depicts a schematic flowchart of a sampling method of the invention. (5) Flow (breath) inlet. (6) Flow outlet.
Figure 3. Schematic representation of desorption of the volatile compounds by heated injector into GC-MS. (7) Coated capillary liner in the injector body. (8) Heated gas chromatography injector. (9) Gas chromatography oven. (10) Mass spectrometer.
Figure 4 depicts exemplary chromatograms (top) and mass-spectra (bottom) of volatile compounds captured from exhaled breath by the method and device of the invention. A) pentane; B) decane; C) acetaldehyde; and D) octane.
Figure 5 depicts the structure of an exemplary sol-gel to be coated in capillary. SUMMARY OF THE INVENTION
The present invention relates to a method and a device for sampling analytes, such as volatile compounds from exhaled air, in particular biomarkers or compounds of toxicological or forensic interest.
Briefly, the analytes, which are typically volatile compounds are initially captured on sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof. Subsequently, the analytes may be released from the surface for analysis with any known suitable method, such as GC-MS or the like.
The present invention relates to the following aspects.
1. A method for collecting and determining the amount of an analyte sample from exhaled breath comprising the steps of:
a. providing a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof; and
b. contacting the exhaled breath with the adsorbent surface whereby the analyte sample to be collected binds to the adsorbent surface, c. determining the amount of said analyte, wherein the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of heat and/or a solvent.
2. The method according to aspect 1 , wherein the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of heat, preferably at 200-300°C. The method according to aspect 1 , wherein the determination step comprises the step (c') of desorbing the bound anaiyte from the adsorbent surface by means of heating from 50-250X.
The method according to any of the preceding aspects, wherein the determination step comprises the step (c') of desorbing the bound anaiyte from the adsorbent surface by means of a solvent such as acetonitrile and/or methanol.
The method according to aspect 1 , wherein the contacting step is conducted during a time span of no more than 1 minute, preferably during less than 50 seconds, 45 seconds, 30 seconds, 20 seconds or 10 seconds.
6. The method according to any of the preceding aspects, wherein the adsorbent surface comprises two compounds selected from the groups consisting of a phenyl methyl silicone, polyethylene glycol and a sol-gel of silica, coated on a first and a second part of the adsorbent surface of the capillary .
The present invention further relates to the following items:
1. A method for collecting an anaiyte sample from exhaled breath, comprising the steps of:
a. providing a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof; and
b contacting the exhaled breath with the adsorbent surface whereby the anaiyte sample to be collected binds to the adsorbent surface.
2. The method according to any of the preceding items, wherein the anaiyte is a volatile compound.
3. The method according to any of the preceding items, wherein the anaiyte is a biomarker for a disease or a condition.
4. The method according to any of the preceding items, wherein the anaiyte is a marker of toxicological interest. The method according to any of the preceding items, wherein the analyte is a marker for a substance of abuse, such as an opiate, a cannabinoid, an
amphetamine, or ***e, or a marker of exposure to environmental or
occupational hazardous chemical, or a marker of exposure to a chemical weapon. The method according to any of the preceding items, wherein the adsorbent surface comprises a compound selected from the groups consisting of a phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.
The method according to any of the preceding items, wherein the analyte to be sampled is selected from the group consisting of pentane, decane, acetaldehyde and octane.
The method according to any of the preceding items, wherein the method is further for determining the amount of an analyte in a sample from exhaled breath, and comprises the step (c) of determining the amount of bound analyte.
The method according to item 8, wherein the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface.
The method according to item 8, wherein the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of heat, preferably at 200-300°C or a solvent such as acetonitrile or methanol.
The method according to any of items 8-10, wherein the determination step comprises the step (c") of determining the analyte by means of GC, LC, MS or a combination of these methods
The method according to item 1 1 , wherein the determination step comprises the step (c") of determining the analyte by means of GC-MS or LC-MS.
The method according to any of the preceding items, wherein the sampling device is a device according to any one of items 15-31.
The method according to any of the preceding items, wherein the contacting step involves transporting the exhaled breath through the capillary by the force of exhalation of the subject from which the breath is exhaled.
A sampling device for collecting an analyte sample from exhaled breath, comprising a capillary with an adsorbent surface coated on the inner surface thereof. 16. The device according to any of the preceding device items, characterized in that the adsorbent surface comprises an adsorbent selected from the group consisting of phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.
17. The device according to any of the preceding device items, wherein the adsorbent comprises phenyl methyl silicone.
18. The device according to any of the preceding device items, wherein the adsorbent comprises polyethylene glycol.
19. The device according to any of the preceding device items, wherein the capillary has an inner cross-sectional dimension of 2-10 mm.
20. The device according to any of the preceding device items, wherein the capillary has an inner cross-sectional dimension of 3-5 mm.
21. The device according to any of the preceding device items, wherein the capillary is 1-10 cm or 3-7 cm in length.
22. The device according to any of the preceding device items, wherein the capillary is 5-10 cm in length.
23. The device according to any of the preceding device items, wherein the capillary is substantially straight.
24. The device according to any of the preceding device items, wherein the capillary comprises one or more bends or is spiral-shaped.
25. The device according to any of the preceding device items, wherein the capillary has a circular or substantially circular inner cross-section.
26. The device according to any of the preceding device items, wherein the capillary has a circular or substantially circular outer cross-section.
27. The device according to any of the preceding device items, wherein the capillary wall has a thickness of 0.001-2 mm, 0.003-0.5 mm, or 0.005-0.1 mm.
28. The device according to any of the preceding device items, wherein the capillary wall comprises a glass, a metal or a plastic, preferably glass.
29. The device according to any of the preceding device items, wherein the adsorbent surface is 100-400 pm in thickness.
30. The device according to any of the preceding device items, wherein the adsorbent surface is 200-300 μητι in thickness. 31. The device according to any of the preceding device items, wherein the device comprises a mouthpiece and/or a handle, preferably a mouthpiece.
32. A method for diagnosis of a disease or condition such as cancer, in particular lung cancer, comprising detecting a biomarker by a method involving a method according to any of items 1-13.
33. A method for toxicological screening, comprising detecting an analyte being a marker for chemical exposure by a method involving a method according to any of items 1-13.
DETAILED DESCRIPTION
Method for collecting a sample
In a first aspect, the present invention relates to a method for collecting an analyte sample from exhaled breath of a subject, comprising the steps of:
a. providing a sampling device comprising an adsorbent surface; and b. contacting exhaled breath from the subject from which the analyte sample is being taken with the adsorbent surface, whereby a sample of the analyte to be collected binds to the adsorbent surface;
characterized in that the sampling device comprises a capillary with an adsorbent surface coated on the inner surface thereof, to which the analyte binds.
The capillary may have any of the properties such as dimensions or materials discussed below under "Properties of the capillary" of the device of the second aspect of the invention. The adsorbent surface may comprise a compound selected from: a phenyl methyl silicone, polyethylene glycol and a sol-gel of silica. Preferably, the adsorbent surface comprises phenyl methyl silicone or polyethylene glycol, as these adsorbents are capable of capturing both hydrophobic and hydrophilic analytes. Both are stable at high temperature (>250°C) and are capable of capturing analytes of interest with different polarity.
The analyte to be sampled may be any analyte present in exhaled breath, capable of binding to the particular adsorbent present in the sampling device. The analyte to be sampled may be a volatile compound. The analyte may be a biomarker, such as a biomarker for a disease or a condition. Preferably, the analyte is a biomarker for cancer, such as lung cancer, or a disease of the respiratory system, such as astma or chronic obstructive lung disease including prodromal stage thereof. The analyte may be a compound of interest for toxicological analysis. For instance, the analyte may be a marker for substance abuse or drug overdose, a marker for exposure to environmental or occupational hazardous substances, a marker for exposure to a chemical weapon, or the like. Preferably, the analyte is selected from pentane, decane, acetaldehyde and octane. The method may further comprise the step (c) of determining the amount of bound analyte. The determination step may comprise the step (c') of desorbing the bound anaiyte from the adsorbent surface. The desorption step (c') may preferably be performed by means of heating the sampling device, preferably at 200-300°C.
In another embodiment, the desorption step (c') may preferably be performed by means of heating the sampling device, at 50-250°C, such as at 50-200X, or at 100 -250°C.
In a presently preferred embodiment, the desorption step (c') is performed by means of heating the sampling device at 100 -250°C. A distinct advantage of the capillary format (see further discussion below under the second aspect) of the sampling device used in the method is that desorption can be conveniently achieved in a standard GC instrument by inserting the capillary into the injector chamber. Alternatively, a solvent such as acetonitrile or methanol can be used for desorption. The solvent containing the desorbed analyte can then proceed to analysis. Preferably, the determination step comprises the step (c") of determining the analyte by means of gas chromatography (GC), liquid chromatography (LC), mass spectrometry (MS) or a combination of these methods. Use of GC-MS or LC-MS is preferred, GC-MS being the most preferred since the capillary can be inserted in the injector for convenient analyte desorption by heating, as discussed above.
The sampling device may be a sampling device according to the second aspect of the present invention described below.
Preferably, the contacting step involves transporting the exhaled breath through the capillary solely, substantially solely, substantially or mainly by the force of the subject's exhalation, without the use of collection reservoirs such as bags or pumps of any kind. In one embodiment, the contacting step is performed for no longer than 2 minutes by the exhaling patient in a time span of no more than approximately 1-2 minutes, such as in a time span of between 1 second-30 seconds, 30 seconds- 1 minute, 1 minute -1.5 minutes, or 1.5 minutes - 2 minutes. I.e. the patient exhales for no longer than approximately 2 minutes, such as in a time span of between 1 second-30 seconds, such as for no longer than 10 seconds, 20 seconds, 30 seconds, 1 minute , 1.5 minutes, or 2 minutes.
In another embodiment, the contacting step can take as little as 30 seconds by the exhaling patient in a time span of no more than approximately 10 - 30 seconds, such as in a time span of between 1-10 seconds, 10 seconds-20 seconds, 20 seconds- 30 seconds.
In a presently preferred embodiment, the patient exhales for less than approximately 10 seconds.
Device for analyte sample collection
In a second aspect, the present invention relates to a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof, characterized in that the adsorbent surface comprises an adsorbent selected from the group consisting of phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.
Preferably, the adsorbent surface comprises phenyl methyl silicone or polyethylene glycol, as these adsorbents are capable of capturing both hydrophobic and hydrophilic anaiytes. Both are stable at high temperature (>250°C) and are capable of capturing anaiytes with different polarity.
The device may optionally comprise a mouthpiece and/or a holder to facilitate handling. In a further aspect, the present invention relates to a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof, characterised in that the adsorbent surface comprises a first polymer immobilized on the first part of the inner surface and a second polymer immobilized on the second part of the inner surface.
Properties of the capillary
The capillary may have an inner cross-sectional dimension of 2-10 mm, preferably 2.5-8 mm, more preferably 2.5-6 mm, most preferably 3-5 mm. The cross-section may have any shape compatible with the application, such as circular, oval, triangular, square, rectangular, pentagonal and octagonal or the like, in most applications however, a substantially circular cross-section is preferable from a practical point of view in particular in terms of handling and availability. The same considerations apply to the outer dimensions of the capillary. It is preferred the outer cross-section is substantially circular to facilitate interfacing with common existing instruments such as GC. However, it is contemplated that in some instances the capillary could be integrated in a larger block or a module, which could be adapted for interfacing with a particular instrument. In such case, the outer dimensions could vary greatly.
The thickness of the capillary wall may be 0.001-2 mm, 0.003-0.5 mm, or 0.005-0.1 mm. The capillary wall thickness is most relevant if easy interface with existing instruments is desired. It is however understood that in cases where the capillary is integrated in a block or a module, there is substantially more freedom in choosing the wall thickness.
The capillary, or the channel therein may be 4-10 cm in length, preferably 4-8 cm, more preferably 6-9 cm, most preferably 7-10 cm. The preferred measures provide a
reasonable compromise between capacity for capture of the adsorbent surface and pressure needed to move the sample through the device.
The capillary is preferably straight, as this provides advantages in terms of storage (In a box), general handling, sampling (lowest possible back pressure) and in particular interface with standard analytical instruments such a GC. Most instruments adapted for samples in capillary are designed on the premise that capillaries are straight, having essentially circular cross-section and having a certain external diameter (about 0.5-1.5 mm) and having a certain length (about 5 cm). It is advantageous to adhere to designs adapted to suit the instruments intended to be used for downstream analysis. However, it is contemplated that in some instances the capillary may also be curved, having one or more bends, or be of a spiral shape. This applies in particular in cases discussed above where the capillary is integrated in a block or a module.
Preferably, the device is arranged such that the exhaled breath can be transported through the device by the force of the subject's exhalation, eliminating the need for collection reservoirs such as bags or pumps of any kind. For the adsorption directly from the exhaled breath to be practicable, the capillary may not be too narrow, too winded or too long, since otherwise the pressure needed to transport a sufficient volume of exhaled breath during the short time that the subject is able to exhale becomes too large. On the other hand, too short or too wide capillaries do not provide unsufficient adsorption capacity.
The structural capillary wall may be made of heat-tolerant inert material such a glass, a ceramic or a metal, allowing for both heat-based and solvent-based desorption. For solvent-based dsorption a wider range of materials is acceptable, including inert plastics such as polyamides, poly(ether etherketone) (PEEK), poly(tetrafluoroethylene) (PTFE), polypropene or polyethylene.
The adsorbent surface may be 100-4QGpm in thickness, more preferably 200-300 μνα. Manufacture of device
The capillary to be coated may be pre-treated, for instance a glass capillary surface may be activated by acid/base treatment. The polymer to be coated is dissolved in a volatile solvent, such as acetone or dichloromethane. Capillary to be coated may then be filled with the polymer solution and left until the solvent has evaporated. The capillary may then be subjected to a heat treatment.
Thus, in summary the present invention provides the advantages of a) ease of use, b) low cost, c) fast (possible to get results at the same day); d) potential for being used as screening method, e.g. for early stage lung cancer.
The term comprising is to be interpreted as including, but not being limited to. All references are hereby incorporated by reference.
EXAMPLES
The following examples are not to be interpreted as limiting.
Example 1 : Manufacture of devices for analyte sample collection
Preparation of capillaries coated with phenyl methyl silicone or poly ethylene glycol The capillary inside surface of a glass capillary was activated by acidic and basic solution before the immobilization of the liquid polymer (phenyl methyl silicone or poly ethylene glycol). For the activation of capillary surface, i.e. to produce reactive silanol groups, the capillary was filled with strong acid such as hydrochloric acid [0.1 M] and left for 60 minutes at room temperature. After 60 min the capillary was rinsed with Milli-Q water and was left to dry. After drying, the capillary was filled with sodium hydroxide [1 M] for 60 minutes to activate the siianol groups (hydroxy! groups, OH). After an additional rinse with Milli-Q water, the capillary was left to dry.
The polymer [phenyl methyl silicone (OV 17 from Sigma-Aldrich) or PEG 400 (Sigma- Aldrich)] to being used for coating was dissolved in acetone or dichloromethane to a concentration of about 2.0 mg/mL to facilitate the coating procedure. The activated capillary was filled with the polymer solution and was left for some hours at room temperature for evaporating of the acetone or dichloromethane. The capillary was then left over night at 40 degrees. The next day the capillary was heated-treated by heating from 40 degrees up to 250 degrees and left for 2 hours at 250 degrees.
The preparation of sol gel-coated capillary
1 .0 mL of tetraethyl orthosilicate (TEOS) and 1.0 mL (3-aminopropyl)triethoxysilane (APTES) were mixed and then 200 mg of polyethylene glycol (PEG) was added and the solution was vortexed for a few seconds. The mixture was then sonicated for 30 minutes in ultrasound bath at room temperature. Subsequently, 100 uL of trifluoroacetic acid (TFA) and 0.8 mL acetonitriie were added and mixed. Finally, 50 uL pure water added to the mixture thus producing a final mixture being a sol-gel solution. All chemicals and reagents can be obtained from Sigma-Aldrich in Sweden.
The capillary was activated as mentioned above and then was filled with sol-gel solution and left at room temperature for 24 hours. After that the capillary was incubated at 55°C for 36 hours.
Finally, the capillary treated with a heat gradient: at 100°C for one hour, then at 50°C for one hour and lastly at 200°C for one hour. After the heat gradient the capillary is ready for use. Figure 5 exemplifies a sol-gel coated capillary. Example 2: Volatile compound collection from exhaled breath
Breath collection: All patients and controls were instructed to not eat anything 4 hours before the breath test. Each study subject performed tidal breathing of unfiltered room air for some minutes. During this time, they inhaled through their nose and exhaled through their mouth into the coated capillary obtained according to Example 1. There were no flowers or plants in the rooms, so the VOCs (volatile compounds) in the breath were considered as originating from the subjects themselves. The breath samples were stored in room temperature.
Example 3: Volatile compound determination from collected samples
The capillary from Example 2 with collected volatile compound was inserted into a GC injector chamber. The injector was programmed heated from 50°C to 250°C. The adsorbed analytes were thermally desorbed into a GC column separation and then detected identified by mass spectrometry. Results for detection of certain volatile compounds is shown in Figure 4A-D.

Claims

A method for collecting and determining the amount of an analyte sample from exhaled breath comprising the steps of: a. providing a sampling device comprising a capillary with an adsorbent surface coated on the inner surface thereof; and b. contacting the exhaled breath with the adsorbent surface whereby the analyte sample to be collected binds to the adsorbent surface, c. determining the amount of said analyte, wherein the determination step
comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of heat and/or a solvent.
The method according to claim 1 , wherein the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of heat, preferably at 200-300°C.
The method according to claim 1 , wherein the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of heating from 50-250°C.
The method according to any of the preceding claims, wherein the determination step comprises the step (c') of desorbing the bound analyte from the adsorbent surface by means of a solvent such as acetonitrile and/or methanol.
The method according to any of the preceding claims, wherein the contacting step involves transporting the exhaled breath through the capillary by the force of exhalation of the subject from which the breath is exhaled.
The method according to claim 5, wherein the contacting step is conducted during a time span of no more than 1 minute, preferably during less than 50 seconds, 45 seconds, 30 seconds, 20 seconds or 10 seconds.
The method according to any of the preceding claims, wherein the adsorbent surface comprises a compound selected from the groups consisting of a phenyl methyl silicone, polyethylene glycol and a sol-gel of silica.
The method according to any of the preceding claims, wherein the adsorbent surface comprises two compounds selected from the groups consisting of a phenyl methyl silicone, polyethylene glycol and a sol-gel of silica, coated on a first and a second part of the adsorbent surface of the capillary .
9. The method according to any of the preceding claims, wherein the analyte to be sampled is selected from the group consisting of pentane, decane, acetaldehyde and octane.
10. A sampling device for collecting an analyte sample from exhaled breath, comprising a capillary with an adsorbent surface coated on the inner surface thereof.
11. The sampling device according to any of the preceding claims, wherein the capillary is 1-10 cm or 3-7 cm in length.
12. The sampling device according to any of the preceding claims, wherein the capillary has an inner cross-sectional dimension of 2-10 mm or 3-5 mm.
13. The sampling device according to any of the preceding claims, wherein the capillary wall has a thickness of 0.001-2 mm, 0.003-0.5 mm, or 0.005-0.1 mm.
14. The sampling device according to any of the preceding claims, wherein the adsorbent surface is 100-400 μΜ or 200-300 μ in thickness.
15. A method for diagnosis of a disease or condition such as cancer comprising a method according to any of claims 1-9.
16. A method for toxicological analysis from exhaled breath according to any of claims 1-
9.
PCT/SE2016/050258 2015-03-30 2016-03-29 Method and device for analyte sampling and analyte concentration determination from exhaled breath WO2016159863A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE1550376-6 2015-03-30
SE1550376A SE1550376A1 (en) 2015-03-30 2015-03-30 Method and device for analyte sampling from exhaled breath

Publications (1)

Publication Number Publication Date
WO2016159863A1 true WO2016159863A1 (en) 2016-10-06

Family

ID=55949047

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE2016/050258 WO2016159863A1 (en) 2015-03-30 2016-03-29 Method and device for analyte sampling and analyte concentration determination from exhaled breath

Country Status (2)

Country Link
SE (1) SE1550376A1 (en)
WO (1) WO2016159863A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024073834A1 (en) * 2022-10-07 2024-04-11 Cannabix Technologies Inc. Apparatus and methods for capturing non-volatile and semi-volatile substances from breath

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150064796A1 (en) * 2013-08-28 2015-03-05 University Of Louisville Research Foundation, Inc. Noninvasive detection of lung cancer using exhaled breath

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150064796A1 (en) * 2013-08-28 2015-03-05 University Of Louisville Research Foundation, Inc. Noninvasive detection of lung cancer using exhaled breath

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
LOURENCO; TURNER, METABOLITES, vol. 4, 2014, pages 465 - 498
N. PELED ET AL: "Abstract A36: Breath Biomarkers in the Post NLST-era for the Discrimination between Malignant from Benign Pulmonary Nodules", CLINICAL CANCER RESEARCH, vol. 18, no. 3_Supplement, 1 February 2012 (2012-02-01), pages A36 - A36, XP055153214, ISSN: 1078-0432, DOI: 10.1158/1078-0432.12AACRIASLC-A36 *
TREFZ ET AL., ANAL BIOANAL CHEM, vol. 405, 2013, pages 3105 - 3115

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024073834A1 (en) * 2022-10-07 2024-04-11 Cannabix Technologies Inc. Apparatus and methods for capturing non-volatile and semi-volatile substances from breath

Also Published As

Publication number Publication date
SE1550376A1 (en) 2016-10-01

Similar Documents

Publication Publication Date Title
JP6332597B2 (en) Portable sampling device and method for sampling drug substance from exhaled breath
ES2627090T3 (en) System and method for elution and substance analysis of an exhaled aerosol sample
CN102498398B (en) System and method for drug detection in exhaled breath
US20210085213A1 (en) Cannabis drug detection device
Kataoka et al. Recent advances in SPME techniques in biomedical analysis
JP2013504074A5 (en)
US11617521B2 (en) Multiple breath sampling method
US9267866B2 (en) Capillary microextractor of volatiles (CMV)
Hu et al. In situ solid phase microextraction sampling of analytes from living human objects for mass spectrometry analysis
JP2016136152A5 (en) Method for analyzing biological chemicals via a sampling device attached to underwear and underwear
JP2016504591A (en) Cancer diagnosis method using respiratory gas
US20190307396A1 (en) Device and method for detection of cannabis and other controlled substances using faims
CN104939831B (en) The system and method and application thereof of sample are collected from the expiratory air of subject
CN106796217A (en) For the composition of directly breathing sampling
TW201445138A (en) Apparatus and method for analyzing breath gas mixture for halitosis detection
WO2016159863A1 (en) Method and device for analyte sampling and analyte concentration determination from exhaled breath
Kang et al. Gas concentrator with an activated carbon adsorbent for breath analysis
RU2363381C1 (en) Method of detecting oncological disease of lungs
EP4319629A1 (en) System and method for breath sampling and breath analysis
CN203249801U (en) Gas sampling enrichment device based on nano material
Azzouz et al. Recent Advances in Targeting Clinical Volatile Organic Compounds (VOC)
JP2024515320A (en) Gas or breath sampling device that captures both aerosol and vapor fractions
JP6256986B2 (en) Evaluation method of esophageal cancer
WO2024073834A1 (en) Apparatus and methods for capturing non-volatile and semi-volatile substances from breath
JP2018532997A (en) Multi-bed sorbent tube and its use

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16721259

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC , EPO FORM 1205A DATED 220118.

122 Ep: pct application non-entry in european phase

Ref document number: 16721259

Country of ref document: EP

Kind code of ref document: A1