CN116297795A - Analysis method for detecting concentration of acetone in exhaled end gas - Google Patents
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Abstract
The invention is an analysis method for detecting the concentration of acetone in the expired air, the invention collects the expired air of a patient into an air bag through noninvasive expired air analysis detection, and carries out sampling semiconductor refrigeration on-line dehumidification detection on the expired air of the patient, thus realizing high-selectivity sensitive detection, the response time is 80ms, the quantitative range is 30ppbv-325ppbv, the detection limit is 2.6ppbv, and compared with the conventional blood sampling detection for the blood sugar level of diabetes, the analysis method has the advantages of noninvasive property, high sensitivity, high detection speed, no need of any sample pretreatment and the like. The exhaled breath acetone is used as a specific marker and a characterization index of a diabetic patient, the exhaled breath acetone concentration of the diabetic patient is obviously higher than that of a normal person, and the analysis and detection method has important clinical scientific research reference value for early diagnosis of the diabetic patient and evaluation of blood sugar concentration.
Description
Technical Field
The invention belongs to the field of analytical chemistry instrument detection, and particularly relates to an analysis method for detecting the concentration of acetone in exhaled end gas.
Background
Diabetes is a chronic disease that is not easily diagnosed in the early stages of onset, but is not found until after complications are caused, and is difficult to treat in the later stages. Blood glucose is detected by blood glucose test paper, the blood is attached to the test paper by puncturing the finger, earlobe and other parts, and the color change of the test paper is observed and compared with the standard color to roughly estimate the blood glucose value.
There are thousands of components in exhaled breath, acetone as the primary volatile organic compound in exhaled breath, and it plays an extremely important role in the diagnosis of diabetes. Acetone as a biomarker, diabetes can be diagnosed by measuring the acetone content of exhaled breath, sweat or urine. Studies have shown that diabetics, due to their congenital defects or lack of insulin mechanism in the body, under conditions of insufficient glucose storage in the body, need the liver to metabolize fatty acids as additional energy supplements, during which process ketones such as acetone are produced, which is expelled from the body along with blood circulation via the lungs, urine or sweat. And the acetone content of the exhaled breath is positively correlated with the blood sugar content of diabetics, thus providing possibility for diagnosing diabetes by detecting the acetone content of the exhaled breath.
The existing detection method of the exhaled breath acetone comprises a gas chromatography, a photometry, a sensor, other methods and the like, wherein the chromatography and chromatography-mass spectrometry technology is most widely applied to acetone detection, but has the defects of higher cost, large volume, special technician operation and the like, the photometry has simple operation steps, but has poor stability and reproducibility, and the sensor method has the advantages of microminiaturization, short response time and the like, but has inaccurate quantification and the like.
Huang Honghu et al discloses a nano sensor for early diagnosis of diabetes and a preparation method thereof (patent number ZL 200310108212.3) which is prepared from Ag + The ZSM-5 sensitive film is combined with the piezoelectric resonant mass sensor QCM. The acetone gas response of the sensor to the characterization index of the diabetes is used for detecting the concentration of acetone molecules in the exhaled air of the human body and is used as an auxiliary means for diagnosing the diabetes and a reference of an early-stage condition self-diagnosis system. However, the sensor has the defects of long response time, low sensitivity, poor quantitative accuracy and the like.
Sun Yane A blood glucose meter based on acetone sensor and its detection method (patent No. CN 201810507071.9) comprise a sensor array detection unit for detecting acetone concentration of exhaled air in user's mouth, converting the detected acetone gas concentration data into electric signal, transmitting to a signal acquisition processing circuit, and arranging ZnFe in the sensor array detection unit 2 O 4 And MoS 2 The acetone sensor of the phase doped sensitive film, the signal acquisition processing circuit is used for filtering and amplifying the electric signal output by the sensing array detection unit, and the singlechip microprocessor unit is used for determining the blood sugar content of a user. The detector and the detection method mainly introduce a signal processing algorithm, do not have detection spectrograms and quantitative detection concentration of the exhaled breath acetone and the like, and do not have practical case detection results.
Sun Jingxi et al, which discloses a rapid detecting device for diabetes mellitus for endocrinology (patent number CN 201910233369. X), comprises a urine analyzer, an acetone detector and a blood sugar detector, and can detect the concentration and content of acetone in urine and exhaled air of a patient and the blood sugar of the patient, and the detected data can be seen through a display screen, but the specific analysis and detection method and diagnosis results of exhaled air acetone are not introduced, and the practical clinical application value is yet to be considered.
Aiming at various problems existing in the analysis and detection method, the invention adopts the direct photoionization ion mobility spectrometry to detect the acetone and applies the method to the detection of the acetone content in the exhaled air of a human body, compares the acetone concentration of the exhaled air of a normal person and a diabetic patient, and finds that the acetone concentration of the exhaled air of the diabetic patient is obviously higher than that of the exhaled air of the normal person.
Disclosure of Invention
The technical problems to be solved by the invention are as follows: the method solves the problems of monitoring the acetone concentration of the exhaled end gas and diagnosing the diabetes patients, solves the problems of high humidity (100%RH) in the exhaled gas and the influence of other components in the exhaled gas on the trace low-concentration ppbv acetone in the exhaled gas, realizes the collection of the acetone sample of the exhaled end gas and the online refrigeration and dehumidification detection of a semiconductor, has the analysis and detection time of only 80ms, has the detection limit of 2.6ppbv, can better reflect the acetone concentration of human alveolar blood exchange, and establishes an analysis method for early pre-bed diagnosis of the diabetes patients by detecting the acetone concentration of the exhaled end gas, has the advantages of noninvasive and noninvasive performance, can quickly obtain the detection result, avoids the difficulty of blood sampling and detection of the diabetes patients, and is easy to clinically popularize in a large scale.
The specific contents include:
an analytical method for detecting the concentration of acetone in exhaled breath, which is characterized in that: firstly, collecting an expired air, namely alveolar air (air expired by maximum force after air is inhaled and exhausted in a first effort mode) sample of a person in a Tesla air bag, and detecting the expired air sample in the collected sampling bag by a sampling pump and an on-line semiconductor refrigerating and dehumidifying process and then entering an instrument; the acetone exhaled by the ion mobility spectrometry is photoionization by a vacuum ultraviolet lamp, and acetone (Ac) is generated after photoionization 2 ) Product ion peak Ac of (2) 2 H + The product ions migrate to reach the Faraday disc in a positive high-pressure mode in the migration tube, and the signal intensity is obtained through detection; from this peak intensity, the concentration of exhaled end air acetone was calculated.
The method comprises the steps that an expired terminal gas sample is collected in a Tesla gas bag, an adopted device consists of a disposable air blowing nozzle, a glass breathing pipe, an expired gas indicator lamp, a gas flow sensor, a three-way two-position electromagnetic valve and a glass gas outlet pipe, the glass air blowing pipe is connected with the Tesla sampling gas bag in a sealing way, the expired gas indicator lamp is manually pressed slightly before expiration each time, the sampling device is started, and when the lamp flashes in red left and right, the air blowing can be indicated; then the sampled person uses disposable air blowing nozzle, when the expired air passes through the air flow sensor arranged on the breathing tube from the air blowing nozzle, the flow speed is tracked in real time, when the flow speed of the blown air is larger than the set threshold value, the signal of the sensor is transmitted to the electromagnetic valve, the electromagnetic valve switches the air path, the front end air expired by the sampled person and smaller than the set value of the flow speed is discharged, the air at the expired end is collected into the Teddar air bag, and the sampled air bag gradually becomes larger and is filled after 1 or more times of blowing.
The photoelectric ion mobility spectrometry works in a positive ion detection mode, the temperature of a mobility tube is set at 20-150 ℃, the voltage between rings is set at 200-350V, a sample of exhaled terminal gas in a Tesla sampling air bag is sampled by a sampling pump of the ion mobility spectrometry, a flowmeter controls the sampling flow rate, the flow rate is constant (20-200 ml/min), the gas enters the ion mobility tube through a rear end gas inlet of the mobility tube after being dehumidified by semiconductor refrigeration on line, the gas is distributed in the whole mobility region and the reaction region of the mobility tube, the gas is ionized into product ions of sample molecules in the ionization region by photoelectric ions, the product ions of exhaled gas acetone reach a detector under the action of an electric field in the mobility region, a current signal is converted into a voltage signal, the signal intensity of exhaled gas acetone is detected, and the signal intensity of exhaled gas acetone is substituted into a quantitative equation of exhaled gas acetone to obtain the concentration of exhaled gas acetone; the quantitative curve is a quantitative curve which is fit and drawn by adopting the signal intensity of acetone ion peaks detected by different known acetone concentration exhaled breath samples, and the obtained quantitative equation y=a×exp (-C/b) +y 0 Wherein a= -949, y 0 ＝656，b＝65。
The expired end gas samples were tested at room temperature (26 ℃) and at the semiconductor on-line cool dehumidified temperatures (-20 ℃ to-50 ℃), respectively.
The human normal healthy person and the diabetic patient.
The invention adopts the single photon ion mobility spectrometry on-line detection technology of direct photoionization, has very high sensitivity and specific signal response to acetone in the exhaled breath of a person, and can obtain the concentration of the exhaled breath acetone in real time. According to the invention, rapid clinical diagnosis is carried out through noninvasive exhaled breath analysis and detection and through specific marker acetone in exhaled breath of a diabetic patient, the exhaled breath of the patient is firstly collected into an air bag and sampled and detected, high-selectivity sensitivity detection is realized, the response time is 80ms, the quantitative range is 100ppbv-325ppbv, compared with the conventional blood sampling detection for detecting the blood sugar level of the diabetes, the analysis method has the advantages of noninvasive, high sensitivity, high detection speed, no need of any sample pretreatment and the like, has very strong specificity for the diagnosis of the diabetes patient, does not need to sample detection, and is very suitable for the pre-bed diagnosis of the diabetes. The exhaled breath acetone is used as a specific marker and a characterization index of a diabetic patient, the exhaled breath acetone concentration of the diabetic patient is obviously higher than that of a normal person, and the analysis and detection method has important clinical scientific research reference value for early diagnosis of the diabetic patient and evaluation of blood sugar concentration.
Drawings
Fig. 1 is a schematic structural diagram of an analysis method for detecting the concentration of acetone in exhaled end gas, wherein 1 is a disposable air blowing nozzle, 2 is an exhaled air flow rate indicator lamp, 3 is an exhaled air flow rate sensor, 4 is an exhaled air blowing pipe, 5 is a two-way three-position electromagnetic valve, 6 is a disposable tidela sampling air bag, 7 is a semiconductor refrigeration on-line dehumidification device, 8 is a photoionization positive ion mobility spectrometry, 9 is a mass flowmeter, and 10 is a gas sampling pump.
FIG. 2 is an ion mobility spectrum of exhaled end air acetone concentration at different mobility tube temperatures
FIG. 3 is a graph of the quantification of exhaled breath acetone concentration
FIG. 4 is a graph showing the ion mobility profile of normal person exhaling end gas acetone at room temperature and after semiconductor refrigeration and dehumidification
FIG. 5 is a graph showing ion mobility profile of exhaled end air acetone at moderate temperature and after semiconductor refrigeration and dehumidification for diabetics
Detailed Description
Example 1
An analysis method for detecting the concentration of acetone in the exhaled end gas comprises the steps of firstly, collecting an exhaled air sample of a person in a Tesla air bag, and enabling the collected exhaled air sample in the sampling bag to enter an instrument through a sampling pump for detection;
as shown in figure 1, the device adopted in the tedlar gas bag for collecting the exhaled end gas sample consists of a disposable mouthpiece, a glass breathing tube, an exhaled gas indicator lamp, a gas flow sensor, 1 two-way three-position electromagnetic valve and a glass gas outlet tube, wherein the glass gas outlet tube is hermetically connected with the tedlar sampling gas bag, the exhaled gas indicator lamp is manually pressed down before each exhalation, the sampling device is started, and when the lamp flashes in red left and right, the gas can be blown; then the sampler uses a disposable air blowing nozzle, when the exhaled air passes through a flow sensor arranged on a breathing tube from the air blowing nozzle, the flow velocity of the exhaled air is tracked in real time, when the flow velocity of the exhaled air is larger than a set threshold value, a signal of the sensor is transmitted to an electromagnetic valve, the electromagnetic valve switches an air path, the front-end air exhaled by a subject and smaller than the set flow velocity value is exhausted, the tail-end air in the stage of exhalation is collected into a Teddy air bag, and the sampled air bag gradually becomes larger and is filled after being inflated for a plurality of times. The acetone exhaled by the ion mobility spectrometry is photoionization by a vacuum ultraviolet lamp, and acetone (Ac) is generated after photoionization 2 ) Product ion peak (Ac) 2 H + ) The product ions migrate to a Faraday disc in a positive high-pressure mode in a migration tube, and the signal intensity is obtained through detection; from this intensity, the concentration of exhaled breath acetone was calculated.
Example 2
The apparatus and procedure used was the same as in example 1, except that: the method comprises the steps of directly detecting acetone at the tail end of an exhale through single photon photoionization, wherein the analysis response time is 80ms, the subject exhales through an exhaled tail end gas collecting device at a constant blowing flow rate of 5L/min, and samples of the tail end gas of the exhaled part are collected into a Tedella bag, and a semiconductor on-line refrigerating and dehumidifying detection is carried out after a subsequent instrument is used for sampling.
The photoelectric ion mobility spectrometry works in a positive ion detection mode, the temperature of a mobility tube is set at 50-130 ℃, the voltage between rings is set at 200-350V, an expired air sample in a Tesla sampling air bag is sampled by a sampling pump of the ion mobility spectrometry, a flowmeter controls the sampling flow rate (20-200 ml/min), the expired air sample enters the ion mobility tube from an air inlet at the rear end of the mobility tube at a constant flow rate, the expired air sample is distributed in the whole mobility region and the reaction region of the mobility tube, and is ionized into product ions of sample molecules by photoelectric ions in an ionization region, the product ions of expired air acetone reach a detector under the action of an electric field in the mobility region, the current signal is converted into a voltage signal, and the ion mobility spectrogram of expired air acetone is detected. As shown in fig. 2, the transition times and signal intensities of the product ion peaks of the exhaled end-gas acetone were examined at different transition tube temperatures of 50 ℃, 60 ℃, 70 ℃, 80 ℃, 90 ℃, 100 ℃, 110 ℃, 120 ℃ and 130 ℃, the product ion peaks of the exhaled end-gas acetone were gradually advanced and the ion loss decreased signal intensity was gradually increased as the transition tube temperature was increased.
Example 3
The apparatus and procedure used was the same as in example 1, except that: an analysis method for detecting the acetone concentration of the exhaled end gas for diagnosing diabetes comprises preparing the acetone mother gas concentration of the exhaled end gas to 6.8ppm, obtaining an acetone sample (100%RH) of 33-320ppbv by changing the flow rate of the dilution gas within the flow rate range of 1L-10L mL/min, detecting the acetone of the exhaled end gas with different concentrations by pumping and sampling at a certain flow rate at the tail end of an ion migration spectrum, obtaining the ion migration spectrum of the exhaled end gas acetone, obtaining the average value of signal intensity by five sample injection analysis, establishing a quantitative equation of the acetone signal intensity of the exhaled end gas and the acetone concentration of the exhaled end gas, the end acetone samples were expired at different concentrations of 33.8ppbv, 37.6ppbv, 42.2ppbv, 48.2ppbv, 51.9ppbv, 56.2ppbv, 61.3ppbv, 67.3ppbv, 74.7ppbv, 83.9ppbv, 95.8ppbv, 111.5ppbv, 133.3ppbv, 144.7ppbv, 165.9ppbv, 194.3ppbv, 219.4ppbv, 272ppbv, 323.8ppbv, and the signal intensities of the acetone ion peaks detected for the end gas samples were 88.5mV, 108.5mV, 165.7mV, 198.5mV, 215.5mV, 237.1mV, 267.3mV, 302.6mV, 360.4mV, 389.3mV, 429.8mV, 478.9, 529.3, respectively,543.9mV, 572.4mV, 592.9mV, 618.1mV, 638.7mV and 649.5mV, fitting the acetone concentration to the detected acetone flat intensity to obtain a quantitative equation of y=a.exp (-C/b) +y 0 Wherein a= -949, y 0 =656, b=65, the quantitative range is 30ppbv-325ppbv, and the limit of detection (LOD, S/n=3) is 2.6ppbv, as shown in fig. 3.
Example 4
The apparatus and procedure used was the same as in example 1, except that: an analytical method for detecting the concentration of acetone in the end gas of exhalation for diagnosing diabetes mellitus, the temperature of a migration tube is set to be 100 ℃, and the drift gas flow rate is 500ml/min. Collecting a normal person exhaled end gas sample in a Tesla sampling bag, after the exhaled end gas sample is subjected to semiconductor refrigeration and dehumidification, detecting the exhaled end gas acetone by pumping and sampling at a certain flow rate at the tail end of an ion mobility spectrometry and entering a mobility tube, and obtaining an ion mobility spectrometry chart of the normal person exhaled end gas acetone, wherein the ion mobility spectrometry chart is shown in fig. 4, and software automatically reads and obtains the signal intensity of the exhaled end gas acetone. Substituting the signal intensity of the acetone of the exhaled end gas into a quantitative equation of the acetone of the exhaled end gas to obtain the concentration of the acetone of the exhaled end gas, which is 56ppbv. The test result proves that the signal intensity and the sensitivity of the acetone detection of the exhaled end gas are obviously improved by more than 2 times after the semiconductor is subjected to on-line refrigeration and dehumidification.
Example 5
The apparatus and procedure used was the same as in example 1, except that: an analytical method for detecting the concentration of acetone in the end gas of exhalation for diagnosing diabetes mellitus, the temperature of a migration tube is set to be 100 ℃, and the drift gas flow rate is 500ml/min. Collecting a sample of the tail end of the exhaled breath of a diabetic patient in a Tesla sampling bag, performing detection on acetone of the tail end of the exhaled breath by pumping and sampling the sample of the tail end of the exhaled breath at a certain flow rate after the sample of the exhaled breath is not subjected to semiconductor refrigeration and dehumidification and is subjected to semiconductor refrigeration and dehumidification, and detecting acetone of the exhaled breath at a certain flow rate to obtain an ion migration spectrogram of acetone of the exhaled breath, wherein as shown in fig. 5, software automatically reads and obtains the acetone signal intensity of the exhaled breath. Substituting the signal intensity of the acetone of the exhaled end gas into a quantitative equation of the acetone of the exhaled end gas to obtain the concentration C of the acetone of the exhaled end gas to be 310ppbv. The test result proves that the signal intensity and sensitivity of the acetone detection of the exhaled end gas are obviously improved by 2-3 times after the semiconductor is subjected to on-line refrigeration and dehumidification. In addition, the test results show that the acetone concentration of the exhaled end gas of the diabetic patient detected after dehumidification and not dehumidification is higher than that of the exhaled end gas of a normal person, and the acetone concentration of the exhaled end gas can be used for diagnosis of the diabetic patient.
Claims (5)
1. An analytical method for detecting the concentration of acetone in exhaled breath, which is characterized in that: firstly, collecting an expired air, namely alveolar air (air expired by maximum force after air is inhaled and exhausted in a first effort mode) sample of a person in a Tesla air bag, and detecting the expired air sample in the collected sampling bag by a sampling pump and an on-line semiconductor refrigerating and dehumidifying process and then entering an instrument; the acetone exhaled by the ion mobility spectrometry is photoionization by a vacuum ultraviolet lamp, and acetone (Ac) is generated after photoionization 2 ) Product ion peak Ac of (2) 2 H + The product ions migrate to reach the Faraday disc in a positive high-pressure mode in the migration tube, and the signal intensity is obtained through detection; from this peak intensity, the concentration of exhaled end air acetone was calculated.
2. The method of analysis according to claim 1, wherein: the method comprises the steps that an expired terminal gas sample is collected in a Tesla gas bag, an adopted device consists of a disposable air blowing nozzle, a glass breathing pipe, an expired gas indicator lamp, a gas flow sensor, a three-way two-position electromagnetic valve and a glass gas outlet pipe, the glass air blowing pipe is connected with the Tesla sampling gas bag in a sealing way, the expired gas indicator lamp is manually pressed slightly before expiration each time, the sampling device is started, and when the lamp flashes in red left and right, the air blowing can be indicated; then the sampled person uses disposable air blowing nozzle, when the expired air passes through the air flow sensor arranged on the breathing tube from the air blowing nozzle, the flow speed is tracked in real time, when the flow speed of the blown air is larger than the set threshold value, the signal of the sensor is transmitted to the electromagnetic valve, the electromagnetic valve switches the air path, the front end air expired by the sampled person and smaller than the set value of the flow speed is discharged, the air at the expired end is collected into the Teddar air bag, and the sampled air bag gradually becomes larger and is filled after 1 or more times of blowing.
3. The method of analysis according to claim 1, wherein: the photoelectric ion mobility spectrometry works in a positive ion detection mode, the temperature of a mobility tube is set at 20-150 ℃, the voltage between rings is set at 200-350V, a sample of exhaled terminal gas in a Tesla sampling air bag is sampled by a sampling pump of the ion mobility spectrometry, a flowmeter controls the sampling flow rate, the flow rate is constant (20-200 ml/min), the gas enters the ion mobility tube through a rear end gas inlet of the mobility tube after being dehumidified by semiconductor refrigeration on line, the gas is distributed in the whole mobility region and the reaction region of the mobility tube, the gas is ionized into product ions of sample molecules in the ionization region by photoelectric ions, the product ions of exhaled gas acetone reach a detector under the action of an electric field in the mobility region, a current signal is converted into a voltage signal, the signal intensity of exhaled gas acetone is detected, and the signal intensity of exhaled gas acetone is substituted into a quantitative equation of exhaled gas acetone to obtain the concentration of exhaled gas acetone; the quantitative curve is a quantitative curve which is fit and drawn by adopting the signal intensity of acetone ion peaks detected by different known acetone concentration exhaled breath samples, and the obtained quantitative equation y=a×exp (-C/b) +y 0 Wherein a= -949, y 0 ＝656，b＝65。
4. The method of analysis according to claim 1, wherein: the expired end gas samples were tested at room temperature (26 ℃) and at the semiconductor on-line cool dehumidified temperatures (-20 ℃ to-50 ℃), respectively.
5. The method of analysis according to claim 1, wherein: the human normal healthy person and the diabetic patient.
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