KR20190088662A - Standard Sample and Method for Analysis of Exhaled Breath Gas - Google Patents

Abstract

The present invention relates to sampling effective for the exhalation gas analysis using gas chromatography, a standard sample, and an analysis method of exhalation gas using the same and, more specifically, to an analysis method of exhalation gas using the same, which comprises the steps of: (A) collecting an exhalation sample in a static dilution container; (B) heat-treating the container to vaporize the sample; and (C) taking the vaporized sample and analyzing the same by gas chromatography (GC).

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0002]

TECHNICAL FIELD The present invention relates to a standard sample which is effective in analyzing the pungent gas using gas chromatography and a method of analyzing pungent gas using the same.

Recently, much attention has been paid to the technique of acquiring biometric information by measuring various gases contained in breathing gas. During the respiratory action of the human body, the exhaled gas generated is discharged to the outside of the body through the nasal cavity as well as the oral cavity. When the breathing gas is discharged through the mouth, the breathing gas includes gas generated in the oral cavity of the human body. In the case of tooth diseases, gum disease, and tongue impaction, oral gas is predominantly composed of volatile sulfur compounds such as hydrogen sulfide, methyl mercaptan, and dimethyl sulfide, Methylmercaptan is a physiological phenomenon caused by the back of the stomach, which is a pathological phenomenon caused by gum disease. It is known that the exhalation gas contains several hundred kinds of gas components related to the metabolism and respiration of the human body, and specific gases can be used as a biomarker including health information of living organisms. For example, acetone is associated with diabetes and ammonia is associated with kidney disease. In addition, the measurement of carbon monoxide concentration in exhaled gas is recognized as a method of objectively evaluating the smoking status. Furthermore, efforts to diagnose the disease early by analyzing the exhalation of patients with pneumonia, asthma, lung cancer, or obstructive pulmonary disease, which are considered to be particularly related to exhalation, have recently been attempted.

As described above, by measuring the respective components and their concentrations of specific gases from oral gas or respiratory gas contained in the exhaled breath gas, it is possible to roughly estimate the oral diseases and medical diseases of the human body. Analysis of exhaled gas requires continuous monitoring of respiratory gases due to reasons such as chronic respiratory diseases (chronic obstructive pulmonary disease, asthma, pneumonia, etc.) or prognosis of lung cancer. Unlike computed tomography or x-ray photography, And non-invasive methods have the advantage of not imposing psychological and physical burden on patients. In addition, these diagnostic methods can be used to diagnose primary pathologies in infants and children who have difficulty communicating with patients.

On the other hand, in order to analyze the exhaled gas, a very precise high-sensitivity measuring device is required. Whereas conventional gas sensors sense a specific single gas, electronic nose technology for detecting and quantifying a plurality of gases has been developed by constructing sensors for various gases in an array form, The number and kinds of gases that can be used are limited and the cost is high. Gas chromatography is the most common and precise instrument that can be used for expiratory gas analysis. It can be analyzed for trace amounts of sample. Depending on the analysis conditions, the desired components can be separated and analyzed qualitatively and quantitatively.

In order to analyze the expired gas by gas chromatography, the expired sample should be sampled and processed for analysis. Conventional prior arts mainly focus on expiration analysis using a composite gas sensor such as an electronic nose. Unlike an expiratory flow analyzer using a complex gas sensor capable of directly supplying an expiratory sample to an analyzer as a portable device, In the case of chromatography, since the equipment itself is not portable, it is necessary to present a method for sampling and storing samples and for use in analysis. In addition, several tens to several hundreds of trace gases are mixed in the exhalation gas. Therefore, in order to analyze them, not only the respective components must be efficiently separated but also the baseline is required to be stable.

Patent No. 10-1817752 Published Japanese Patent Application No. 10-2015-0005361 Registration No. 10-0983827 Published Japanese Patent Application No. 10-2016-0047565

An object of the present invention is to provide a standard sample which is effective for the analysis of an expired gas using gas chromatography and an analysis method using the same.

According to an aspect of the present invention, there is provided a method of manufacturing a sample container, comprising the steps of: (A) (B) heat treating the vessel to vaporize the sample; And (C) analyzing the vaporized sample by gas chromatography (GC).

At this time, if the sample of the expiration is collected in the static dilution container under the negative pressure state in the step (A), it is possible to collect the component of the expired gas more efficiently.

Since gas chromatography is a device with low mobility, there is a delay between sample collection and analysis time, rather than analysis of components on the spot along with sampling of expired gas from the sample like electronic nose. Therefore, partial liquefaction can be performed depending on the component of the exhaled gas. Therefore, in the step (B) before the analysis by the gas chromatography, the container containing the exhaled gas is heat-treated so that the sample is completely vaporized. According to the above method, it is possible to analyze the exhaled gas without distortion of the component by a simple process. In the following examples, a 2 L volumetric static dilution vessel was used for the preparation of the standard sample. However, in case of taking the sample from the actual sample, a smaller volume vessel can be used for the sample amount and ease of movement / storage. Of course.

As described above, according to the pungent gas analyzing method of the present invention, sampling is simple and accurate analysis can be performed, and thus it can be usefully used for analysis of pungent gas.

In addition, the standard sample used in the method for analyzing the exhaled gas of the present invention is effective for analyzing the exhaled gas, and can be effectively used for quantifying the exhaled gas.

FIG. 1 is a graph showing oven temperature changes of a GC column over time for the analysis of exhaled gas. FIG.
FIG. 2 is a gas chromatogram for an exhaled gas standard sample. FIG.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these embodiments are merely examples for explaining the content and scope of the technical idea of the present invention, and thus the technical scope of the present invention is not limited or changed. It will be apparent to those skilled in the art that various changes and modifications can be made within the scope of the technical idea of the present invention based on these examples.

[Example]

Example 1: Preparation of standard samples

The 2L static dilution container (Sigma) was washed with ethanol and then dried in an oven at 60 ° C for 1 hour. The dried static dilution bottle was attached with a stopper and a valve for injecting the sample.

10 μl each of Hexanal, 2-Butanone, Heptanal, Decane, Pentanal, Acetone, Undecane, Pentane, Benzaldehyde and 1-Propanol were mixed as standard samples. 10 μl of the mixture was injected into the static dilution bottle through a valve Respectively. After the sample was injected, the static dilution bottle was placed in an oven at 60 ° C to completely vaporize the injected sample.

Example 2: Analysis of pungent gas using a standard sample

Analysis of the pungent gas was carried out using the standard sample prepared in Example 1. The analysis was carried out with gas chromatography-mass spectrometer (GCMS 2010 / GCMS QP2010_Shmadzu) and SH-Rtx ^TM -1 (30m) was used as the GC column.

10 μl of the reference sample vaporized in Example 1 was taken and injected into the inlet of the GC-MS instrument. For the GC-MS, the temperature of the sample inlet was maintained at 250 ° C, the split ratio was 20: 1, and the flow rate of the mobile phase gas helium was maintained at 1.02 mL per minute. The GC column oven temperature was maintained at 30 캜 for 5 minutes, increased to 140 캜 at a rate of 5 캜 per minute, and then maintained at 140 캜 for 5 minutes. 1 is a graph showing the change in oven temperature of a GC column with time.

Mass-to-charge ratio (m / z) was measured from 34 to 300 for mass spectrometry. The chromatograms were obtained for 30 minutes. Peaks were identified by searching the database using the library (WILEY7, NIST21, NIST7) for the peaks observed in the chromatogram. FIG. 2 is a gas chromatogram for a standard sample. Table 1 below shows the results of identification and retention time for each peak.

Claims (5)

(A) collecting an expiration sample in a static dilution vessel;
(B) heat treating the vessel to vaporize the sample; And
(C) taking a vaporized sample and analyzing it by gas chromatography (GC);
And analyzing the atmospheric gas.
The method according to claim 1,
The method of analyzing an insoluble gas according to claim 1, wherein in the step (A), a sample of an expiration gas is collected in a static dilution container under a negative pressure state.
The method according to claim 1,
Wherein the heat treatment temperature in step (B) is 50 to 70 占 폚.
3. The method according to claim 1 or 2,
Wherein at least one sample selected from the group consisting of Hexanal, 2-Butanone, Heptanal, Decane, Pentanal, Acetone, Undecane, Pentane, Benzaldehyde and 1-Propanol is used as a standard sample for analyzing the exhaled gas. Analysis method.
3. The method according to claim 1 or 2,
Wherein the step (C) is performed by GC / MS.

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