CN107607641B - Method for detecting mild and moderate fatigue degrees of civil aviation air traffic controller team - Google Patents
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Abstract
The invention relates to a method for detecting the mild and moderate fatigue degrees of a team of civil aviation air traffic controllers, which comprises the following steps: s1: collecting urine samples of a team of civil aviation air traffic controllers, and then preprocessing the urine samples; wherein the urine samples comprise urine samples before work and urine samples after work; s2: respectively carrying out separation detection on the preprocessed samples by adopting a liquid chromatogram-time-of-flight mass spectrometry; s3: matching and comparing the detection results of the urine samples after the work of the team with the detection results of the urine samples before the work, and screening out the characteristic biomarker; s4: and comparing the screened characteristic biomarkers with the biomarkers related to the fatigue degree of the civil aviation air traffic controller, and judging the fatigue degree of the civil aviation air traffic controller. The method for detecting the mild and moderate fatigue degrees of the civil aviation air traffic controller teams and groups is simple and convenient to operate and high in reliability.
Description
Technical Field
The invention relates to the technical field of biological detection, in particular to a method for detecting the mild and moderate fatigue degrees of a team of civil aviation air traffic controllers.
Background
The history of human Research Fatigue dates back to The first world war centuries and a large-scale ergonomic study on Fatigue in Industrial production, particularly in military supplies, was carried out by The british Industrial Fatigue Research Board (after The Industrial Fatigue Research Board, The Industrial Health Research Board). The research report is profoundly affected and even many results have become common knowledge in the industry. For example, reports suggest reducing lengthy labor hours, improving work environments, adjusting shift systems, etc., relieving employee fatigue, and increasing labor productivity. These recommendations still have guiding effects on our present design of labor time settings, lighting in production plants and office environments, etc., and shift system optimization. The indicator used in this report to detect fatigue is labor productivity.
In world war ii, the aerospace industry, particularly the military aviation industry, is rapidly growing. It has been found that pilots experience sudden and even disabling piloting during the piloting of an aircraft to perform a mission, and it has been proposed to characterize fatigue in terms of reduced operating performance. After war, along with military aviation and civil aviation industry development, long distance flight and flight round the clock become more and more common, and people find that fatigue problem is more complicated, and fatigue can accumulate, forms accumulative fatigue. During this period, fatigue was still studied mainly by ergonomic means.
Along with the increasing attention of people to social safety, the research on the fatigue problem is also expanded from the work efficiency research to the subjects of psychology, neuroscience, sleeping, physiology and biochemistry and the like. The disciplines establish corresponding methods for detecting fatigue, such as the Stanford somnolence scale based on subjective feeling, the visual analogy scale and the like of psychology, and the Wisconsin card classification test (WSCT), the London Tower Test (TOL) and the like based on the cognitive ability for detecting fatigue. Subjective scales are easy to be interfered by emotions and other factors, parameter indexes are difficult to set in cognitive tests, and the ceiling or floor effect is often generated, so that the reliability and the effectiveness of the scales or software are not satisfactory; also, for example, a fatigue detection method based on electroencephalogram change is established in the science of neurology and sleep, and fatigue occurs when delta and theta waves are remarkably increased, and the method is difficult to apply to field detection due to the complexity of electroencephalograph instruments; and trying to establish a corresponding detection method based on physiological reactions such as yawning and eye movement changes such as winking, eyelid closure and the like when the human body is tired. So far, people have little understanding on the biochemical mechanism of fatigue, and the explanation of the biochemical mechanism of fatigue can help develop an objective, stable and rapid method for detecting fatigue based on biochemical indexes.
The method for detecting fatigue by adopting biochemical indexes is a method which is tried by people all the time, such as detecting some hormones, peptides and the like, and researches show that the concentrations of cytosine nucleoside (Cytidine) and adrenocortical esterol (Corticoid) are related to the working pressure; substances related to sleep and biological rhythm, such as Glycopeptides (Glycopeptides), are also detected; but Borbrey et al indicated evidence of the lack of consistency of the data with which these compounds have a correlation with fatigue. In conclusion, although research on the correlation between metabolism of various compounds and fatigue has been attempted from various angles, such as working pressure, sleep, biorhythm, etc., no correlation between any metabolic compound in the human body and fatigue has been found yet.
Metabonomics is a new subject for systematically researching in-vivo metabolic characteristics, and is mainly used for systematically exploring, searching and discovering characteristic indexes, namely biomarkers, of body fluid metabolism of a human body affected by diseases, medicines and the like by detecting and analyzing means such as nuclear magnetic resonance, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry and the like in combination with chemometrics, so that the metabonomics is widely applied to the fields of disease diagnosis, medicine research and development and the like. For example, Mapstone and the like at university of george dun in the united states screen 10 characteristic biomarkers by a metabonomics method of liquid chromatography mass spectrometry (liquid mass analysis) for detecting alzheimer disease, diagnosis can be performed 2-3 years before clinical symptoms of the disease occur, and the accuracy can reach more than 90%; for another example, Naviaux and the like adopt a liquid mass analysis method to research the long-term fatigue syndrome metabolic characteristics, find that male patients have 8 metabolites, female patients have 13 metabolites which are significantly changed, and the analysis result of characteristic working curves (ROC) of the subjects shows that the diagnosis accuracy reaches 94 percent (male) and 96 percent (female); the professor schwann and the like carry out metabonomics research on blood and urine of rats of depression and over-fatigue models by adopting liquid quality analysis, and the result shows that the metabolism of the two groups of rats is abnormally changed compared with that of the control group of rats. The two-dimensional nuclear magnetic resonance spectrometer sampled by Armstrong and the like carries out metabonomic research on the serum of a patient with chronic fatigue syndrome, and the result shows that the content of glutamine and ornithine in the serum of a case group is obviously lower than that of a control group. In the above literature reports, case groups and control groups are used for group comparison for various diseases, but the existing literature methods are not suitable for screening biomarkers related to fatigue degree because fatigue is a recoverable physiological change.
The civil aviation air traffic controller is responsible for commanding safe operation of the aircraft on the air traffic route, and colloquially speaking, the civil aviation air traffic controller is used for commanding the aircraft and comprehensively managing all aircrafts in the whole airspace by referring to information such as radar and the like. During work, controllers often command the operation of a plurality of airplanes simultaneously, and the abilities of concentration, accurate judgment, emergency treatment and the like are needed, so that fatigue is very easy, and the maximum time of each controller for commanding the airplanes in the seats is clear and not more than 2 hours by relevant regulations of the civil aviation bureau. The fatigue produced during the work is mainly caused by mental labor. Chen et al in the literature utilized liquid chromatography-mass spectrometry to screen 3 fatigue-related biomarkers from urine samples of civil aviation air traffic controllers, but did not propose the concept of fatigue degree, nor did they disclose a method for detecting fatigue degree of civil aviation air traffic controllers using fatigue-related biomarkers. The inventor establishes a method for screening human fatigue related biomarkers in the early period. On the basis, the inventor discovers through a large number of experiments that the fatigue degree is related to the post safety execution capacity of the civil aviation air traffic controller, so that the fatigue degree detection can provide powerful new technology and new means for the safety risk control of the civil aviation air traffic controller.
Disclosure of Invention
Aiming at the defects in the prior art, the invention establishes a method for detecting the mild and moderate fatigue degrees of the civil aviation air traffic controller team by using a liquid chromatography-flight time mass spectrometry method, which can be used for detecting and evaluating whether the team can be used on duty or not before the team is started, detecting in the team for finding out the fatigue risk and adjusting in time and the like, and provides a new technology and a new method for detecting and monitoring the fatigue degree of the civil aviation air traffic controller, controlling the fatigue risk and guaranteeing the safe operation of the civil aviation.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
the invention provides a method for detecting the mild and moderate fatigue degrees of a team of civil aviation air traffic controllers, which comprises the following steps: s1: collecting a urine sample of a civil aviation air traffic controller, and then preprocessing the urine sample; wherein the urine samples comprise urine samples before work and urine samples after work; s2: respectively carrying out separation detection on the preprocessed samples by adopting a liquid chromatogram-time-of-flight mass spectrometry; wherein the liquid chromatography can adopt polar HILIC chromatographic column and low-polar C18Performing separation detection on one or more of a chromatographic column and a nonpolar PFPP chromatographic column; s3: matching and comparing the detection result of the urine sample after work with the detection result of the urine sample before work, and screening a characteristic biomarker; s4: and comparing the screened characteristic biomarkers with the biomarkers related to the fatigue degree of the civil aviation air traffic controller team, and judging the fatigue degree of the civil aviation air traffic controller team.
In step S3, the biomarkers related to fatigue of the civil aviation air traffic controller team include, but are not limited to: urocanic acid (C)6H6N2O2HMDB00301), acetylcytosine (N4-Acetylcytidine, C11H15N3O6HMDB 05923), 5-hydroxytryptophan (5-Hydroxy-L-tryptophan, C11H12N2O3HMDB 00472), Dimethylguanosine (N2, N2-Dimehtylguanosine, C12H17N5O5HMDB 04284), acetanilide (N-Acetylarylamine, C)8H9NO, HMDB 01250) and Alpha-CEHC (C)16H22O4，HMDB 01518)。
Preferably, the detection result of the urine sample of the civil aviation air traffic controller team after work is matched and compared with the detection result of the urine sample of the civil aviation air traffic controller team before work, and the characteristic biomarker is screened; among the characteristic biomarkers related to the fatigue degree of the civil aviation air traffic controller team: and (3) looking up urocanic acid, acetylcytosine, 5-hydroxytryptophan, dimethylguanosine, acetanilide and Alpha-CEHC, and judging that the class of the civil aviation air traffic controller has light or moderate fatigue according to the type and the variation range of the characteristic markers.
Preferably, the detection result of the urine sample of the civil aviation air traffic controller team after working for 2-3 hours in the preferred daytime after working is matched and compared with the detection result of the urine sample of the civil aviation air traffic controller team before working, and if urocanic acid is detected and the content of the urocanic acid is up-regulated to or more than 1.4 times, the civil aviation air traffic controller team can be judged to have slight fatigue.
Preferably, the detection result of the urine sample of the civil aviation air traffic controller team after working for 3-5 hours in the preferred daytime after working is compared with the detection result of the urine sample of the civil aviation air traffic controller team before working in the duty in a matching way, and if urocanic acid, acetylcytosine and 5-hydroxytryptophan are detected in the screened characteristic markers, and the content of the urocanic acid, the acetylcytosine and the 5-hydroxytryptophan is respectively reduced to be more than 1.4 times, 1.3 times and more than 1.4 times, the civil aviation air traffic controller team can be judged to have moderate fatigue.
Preferably, the detection result of the urine sample after the civil aviation air traffic controller team works for 5-8 hours preferably in the daytime is matched and compared with the detection result of the urine sample before the civil aviation air traffic controller team works, and if urocanic acid, acetylcytosine, 5-hydroxytryptophan, dimethylguanosine, acetanilide and Alpha-CEHC are detected in the screened characteristic markers, the content of the first five types of urine samples respectively decreases by more than 1.4 times, more than 1.3 times, more than 1.4 times and more than 1.4 times, and the content of the Alpha-CEHC increases by more than 1.3 times, the civil aviation air traffic controller team can be judged to have moderate fatigue.
In step S1, the urine sample pretreatment includes: melting the urine sample stored at-80 ℃ at 0-4 ℃, centrifuging the urine sample at 4 ℃ and 12000rpm for 5min, and collecting the supernatant after centrifugation; 100 μ L of the supernatant was taken and then diluted with 100 μ L of water.
In step S2, divide intoKeeping the temperature of the sample chamber at 0-4 ℃ in the detection process; when the liquid chromatogram adopts a polar HILIC chromatographic column to separate and detect polar components, the mass spectrometry adopts a positive ion ionization detection mode; liquid chromatography using weakly polar C18When the chromatographic column is used for separating and detecting weak polar and nonpolar components, the mass spectrometry adopts two ionization modes of positive ions and negative ions; when the liquid chromatogram adopts a nonpolar PFPP chromatographic column to separate and detect weak polar and nonpolar components, the mass spectrometry adopts two ionization modes of positive ions and negative ions.
Preferably, in step S2, when the polar HILIC chromatographic column is used for separating and detecting polar components in the liquid chromatography, the mass spectrometry adopts a positive ion detection ionization mode, and the detection conditions are as follows: the mass range is set to 50-1200m/z full scan mode; the optimal capillary voltage of the electrospray ionizer is 3000V, and the cone voltage is 30V; the drying gas is nitrogen, the desolvation flow rate is 800L/h, and the cone gas flow rate is 30L/h; the desolvation temperature is 400 ℃, and the ion source temperature is 100 ℃; leuconeeenkephalin (leucine enkephalin) with the concentration of 0.2ng/mL is used as a mass number calibration internal standard, and the mass of a calibration ion is 556.2771Da ([ M + H ]]+) The liquid phase conditions are UPLC BEHAmide HILIC column (2.1mm × 100mm,1.7 μm), the mobile phase comprises phase A and phase B, the phase A comprises acetonitrile with volume fraction of 95% and aqueous solution with volume fraction of 5% containing 0.1% formic acid, the phase B comprises aqueous solution containing 0.1% formic acid, the column temperature is 40 ℃, the sample injection amount is 2.0 μ L, and the flow rate of the mobile phase is 0.3 mL/min.
Preferably, in step S2, the liquid chromatography employs weakly polar C18When the chromatographic column is used for separating and detecting weak polar and nonpolar components, the mass spectrometry adopts two ionization modes of positive ions and negative ions, and the detection conditions are as follows: the mass range is set to 50-1200m/z full scan mode; the optimal capillary voltage of the electrospray ionizer is 3000V (positive ions) or 2200V (negative ions), and the cone voltage is 30V; the drying gas is nitrogen, the desolvation flow rate is 800L/h, and the cone gas flow rate is 30L/h; the desolvation temperature is 400 ℃, and the ion source temperature is 100 ℃; leuconeeenkephalin (leucine enkephalin) with the concentration of 0.2ng/mL is used as a mass number calibration internal standard, and the mass of a calibration ion is 556.2771Da ([ M + H ]]+) Or 554.2615Da ([ M-H)]-) (ii) a The liquid phase conditions were: UPLC CSH C18column (2.1mm × 100mm,1.7 μm), mobile phase A as aqueous solution containing 0.1% formic acid, phase B as acetonitrile, column temperature 40 deg.C, sample volume 2.0 μ L, and mobile phase flow rate 0.3 mL/min.
Preferably, in step S2, when the liquid chromatography uses a non-polar PFPP chromatographic column to separate and detect weak polar and non-polar components, the mass spectrometry uses two ionization modes, namely positive ions and negative ions, and the detection conditions are as follows: the mass range is set to 50-1200m/z full scan mode; the optimal capillary voltage of the electrospray ionizer is 3000V (positive ions) or 2200V (negative ions), and the cone voltage is 30V; the drying gas is nitrogen, the desolvation flow rate is 800L/h, and the cone gas flow rate is 30L/h; the desolvation temperature is 400 ℃, and the ion source temperature is 100 ℃; leuconeeenkephalin (leucine enkephalin) with the concentration of 0.2ng/mL is used as a mass number calibration internal standard, and the mass of a calibration ion is 556.2771Da ([ M + H ]]+) Or 554.2615Da ([ M-H)]-) The liquid phase conditions are UPLC HSS PFPP column (2.1mm × 100mm,1.7 μm), the mobile phase is aqueous solution containing 0.1% formic acid as phase A, methanol as phase B, the column temperature is 40 deg.C, the sample injection amount is 2.0 μ L, and the flow rate of the mobile phase is 0.3 mL/min.
Preferably, step S3 specifically includes: performing statistical analysis processing on detection data obtained by adopting liquid chromatography-time-of-flight mass spectrometry by adopting data processing software, preferably metabonomics data processing related software, and further preferably performing statistical analysis processing by adopting metabonomics data processing professional software Progenetics QI; the analysis processing method comprises data alignment and peak extraction; the method further comprises the steps of carrying out pairing grouping on the detection result of the urine sample of the civil aviation air traffic controller team after work and the detection result of the urine sample of the civil aviation air traffic controller team before work, then carrying out processing analysis by adopting a partial least square method OPLS-DA, and screening out the characteristic biomarker by taking P less than or equal to 0.05, CV less than or equal to 30%, VIP greater than 1.0 and maximum change multiple more than or equal to 1.2 as screening thresholds.
The technical scheme provided by the invention has the following beneficial effects: (1) the method for detecting the mild and moderate fatigue degrees of the civil aviation air traffic controller team has high reliability of the detection result, can use chromatographic columns with various polarities to detect the same sample for multiple times, can avoid omission on one hand, and can verify the detection method of the chromatographic columns according to the results of simultaneously detecting the same components by different chromatographic columns on the other hand, thereby avoiding the error of a certain chromatographic column and improving the accuracy; (2) civil aviation air traffic controllers often command the operation of multiple airplanes simultaneously during working, and need the capabilities of concentrated attention, accurate judgment, emergency treatment and the like, so that fatigue is very easy to occur; the applicant of the invention discovers through a large amount of experiments that the fatigue degree is related to the post safety execution capacity of a civil aviation air traffic controller; the method for detecting the mild and moderate fatigue degrees can be used for detecting and evaluating whether the civil aviation air traffic controller team can go on duty or not before the team goes on duty, detecting in duty and finding out fatigue risks and adjusting in time and the like; the method is used for evaluating the workload of the civil aviation air traffic controller team, provides a new technology and a new method for detecting and monitoring the fatigue degree of the civil aviation air traffic controller team, controlling the fatigue risk and guaranteeing the safe operation of the civil aviation.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
FIG. 1 is a typical total ion flow diagram of a polar HILIC column positive ion mode detection of a urine sample of a civil aviation air traffic controller in an embodiment of the present invention;
FIG. 2 is a main component analysis score chart of a HILIC column positive ion detection polar component data of a urine sample of a civil aviation air traffic controller in the first embodiment of the invention;
FIG. 3 is a low polarity C of a urine sample of a civil aviation air traffic controller in accordance with an embodiment of the present invention18Detecting a typical total ion flow diagram in a positive ion mode of the column;
FIG. 4 is a C sample of urine from a civil aviation air traffic controller in accordance with one embodiment of the present invention18Detecting weak polar component data principal component analysis score chart by column positive ions;
FIG. 5 shows a schematic diagram of a first embodiment of the present inventionLow polarity C of urine sample of civil aviation air traffic controller18Detecting a typical total ion flow diagram in a column negative ion mode;
FIG. 6 shows a urine sample C of a civil aviation air traffic controller in accordance with an embodiment of the present invention18Detecting weak polarity component data principal component analysis score chart by column negative ions;
FIG. 7 is a typical total ion flow diagram of a nonpolar PFPP column positive ion mode detection of a urine sample of a civil aviation air traffic controller in the first embodiment of the present invention;
FIG. 8 is a plot of the principal component analysis score of the PFPP column positive ion detection non-polar component data of the urine sample of the civil aviation air traffic controller in the first embodiment of the present invention;
FIG. 9 is a typical total ion flow diagram of the nonpolar PFPP column anion mode detection of the urine sample of the civil aviation air traffic controller in the first embodiment of the invention;
fig. 10 is a PFPP column negative ion detection non-polar component data principal component analysis score chart of a urine sample of a civil aviation air traffic controller in the first embodiment of the present invention.
Detailed Description
The technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. The following examples are only for illustrating the technical solutions of the present invention more clearly, and therefore are only examples, and the protection scope of the present invention is not limited thereby. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional store unless otherwise specified.
Example one
1. Volunteer recruitment and sample collection
Volunteer recruitment: 20 volunteers of civil aviation air traffic controllers are recruited in an airport in a country, and the group entering conditions are as follows: healthy, male, no medication, age 20-35 years, with sample collection performed 12 months winter 2014.
Collecting a urine sample: the urine sample is collected once before the volunteer team of the control member goes on duty in 8 hours of work in white duty and is used as a non-fatigue sample, and the urine sample is collected once before the volunteer team of the control member goes off duty in white duty and is used as a fatigue sample. The urine sample is collected by using a sterile urine cup and is subpackaged in a sterile tube for storage. The urine sample is subpackaged in a sterile centrifuge tube and stored at-80 ℃.
2. Pretreatment of urine samples
The urine sample pretreatment step comprises: prior to assay, urine samples stored at-80 ℃ were thawed at 0-4 ℃, centrifuged at 12000rpm for 5 minutes at 4 ℃, and 100. mu.L of the supernatant was diluted with 100. mu.L of water.
3. Analyzing and detecting urine sample by adopting liquid chromatogram-time-of-flight mass spectrometry
The urine sample is detected by using polar chromatographic column HILIC and weak polar chromatographic column C18And the non-polar chromatographic column PFPP is used for separating and detecting polar, weak-polar and non-polar components in the urine respectively.
The liquid phase conditions of a polar HILIC chromatographic column are UPLC BEH Amide HILIC column (2.1mm × 100mm,1.7 mu M), the mobile phase is a water solution with the composition of phase A being 95% acetonitrile and 5% formic acid, the phase B being a water solution with the composition of 0.1% formic acid, the column temperature is 40 ℃, the sample injection amount is 2.0 mu L, the flow rate of the mobile phase is 0.3mL/min, the mass spectrum conditions are that the detection is in a positive ion ionization mode, the mass range is 50-1200M/z full scanning mode, the optimal capillary voltage of an electrospray ionization device is 3000V, the cone voltage is 30V, the dry gas is nitrogen, the desolvation flow rate is 800L/H, the cone gas flow rate is 30L/H, the desolvation temperature is 400 ℃, the ion source temperature is 100 ℃, leucinenkephalin leukaphan (leucine encephalopeptide) with the concentration of 0.2ng/mL is used as the mass internal standard, and the ion mass is 556.2771Da M + M]+)。
Using weak polarity C18The liquid phase conditions of the chromatographic column are as follows: UPLC CSH C18column (2.1mm × 100mm,1.7 μm), mobile phase A phase composition is water solution containing 0.1% formic acid, phase B composition is acetonitrile, column temperature is 40 deg.C, sample amount is 2.0 μ L, mobile phase flow rate is 0.3mL/min, mass spectrum conditions are that two ionization modes of positive ion and negative ion are detected, mass range is set to 50-1200m/z full scanning modeFormula (II) is shown. The optimum capillary voltage for electrospray ionizer was 3000V (positive ions) or 2200V (negative ions), and cone voltage was 30V. The drying gas was nitrogen, the desolvation flow rate was 800L/h, and the cone gas flow rate was 30L/h. The desolvation temperature was 400 ℃ and the ion source temperature was 100 ℃. Leuconeeenkephalin (leucine enkephalin) with the concentration of 0.2ng/mL is used as a mass number calibration internal standard, and the mass of a calibration ion is 556.2771Da ([ M + H ]]+) Or 554.2615Da ([ M-H)]-)。
The liquid phase conditions of a nonpolar PFPP chromatographic column are UPLC HSS PFPP column (2.1mm × 100mm,1.7 mu M), the mobile phase is phase A which is an aqueous solution containing 0.1% formic acid, phase B which is methanol, the column temperature is 40 ℃, the sample volume is 2.0 mu L, the mobile phase flow rate is 0.3mL/min, the mass spectrum conditions are that two ionization modes of positive ions and negative ions are detected, the mass range is set to be 50-1200M/z full scanning mode, the optimal capillary voltage of an electrospray ionizer is 3000V (positive ions) or 2200V (negative ions), the Cone voltage is 30V, the drying gas is nitrogen, the desolvation flow rate is 800L/H, the Cone gas flow rate is 30L/H, the desolvation temperature is 400 ℃, the ion source concentration is 0.2ng/mL, leucineenkephalin (leucine enkephalin) is used as a mass number calibration internal standard, and the calibration ion mass is 556.2771 ([ M + H ] + Da H [ + H [ ([ M + ]]+) Or 554.2615Da ([ M-H)]-)。
Each urine sample is respectively detected by 3 chromatographic columns for polar, weakly polar and nonpolar components, wherein the polar component is only detected in a positive ion mode, and the weakly polar and nonpolar components are detected in a positive ion mode and a negative ion mode, namely each urine sample is detected for 5 times, so that all metabolic compounds in the urine can be detected as far as possible. A blank is inserted into every 10 samples in the detection to prevent cross contamination, and a quality control sample is inserted for quality control. The temperature of the sample chamber was maintained at 4 ℃ during the analytical test.
4. Data processing
And analyzing detection data obtained by detecting the sample by adopting a liquid chromatography/mass spectrometer, and respectively performing statistical analysis processing including data alignment and peak extraction by adopting a metabonomics data processing professional software Progenetics QI. And (3) pairing and grouping the detection result of the urine sample after work and the detection result of the urine sample before work, processing the detection data in the step (3) by adopting an orthogonal partial least squares method (OPLS-DA), and primarily screening the characteristic biomarker by referring to the retention characteristics of the chromatogram and the mass spectrum characteristics according to the condition that P is less than or equal to 0.05, CV is less than or equal to 30%, VIP is greater than 1.0 and the maximum change multiple is greater than or equal to 1.2 as a screening threshold.
Analyzing and screening 5 compounds as characteristic biomarkers in data of 9490 compounds of polar components in urine by using a polar HILIC column positive ion mode; in the presence of weak polarity C18In the data of 10809 compounds as weak polar components in the urine detected by the positive ion mode of the column, 12 compounds are analyzed and screened as characteristic biomarkers; using weak polarity C18In the data of 5247 compounds of weak polar components in the urine detected by a column negative ion mode, characteristic biomarkers meeting conditions cannot be analyzed and screened; analyzing and screening 6 compounds as characteristic biomarkers in the data of 11414 compounds of the nonpolar component in the urine by using a nonpolar PFPP column positive ion mode; in the data of 6176 compounds of nonpolar components in the urine detected by a nonpolar PFPP column negative ion mode, 2 compounds are analyzed and screened as characteristic biomarkers. Specific results are shown in table 1.
5. Comparing the characteristic markers and judging the fatigue degree of the team
Examining the characteristic biomarkers screened in table 1, the biomarkers related to the fatigue degree of the team of civil aviation air traffic controllers were included: urocanic acid (Urocanic acid), acetylcytosine (N4-acylcytidine), 5-hydroxytryptophan (5-Hydroxy-L-tryptophan), dimethylguanosine (N2, N2-Dimehtylguanosine), acetanilide (N-Acetylarylamine), and Alpha-CEHC. Further checking the change multiple of each biomarker, the content of urocanic acid (Urocacic) is reduced by 1.41 times, the content of acetylcytosine (N4-Acetylcytidine) is reduced by 1.39 times, the content of 5-hydroxytryptophan (5-Hydroxy-L-tryptophan) is reduced by 1.42 times, the content of dimethylguanosine (N2, N2-Dimehtylguanosine) is reduced by 1.41 times, the content of acetanilide (N-acetylarylamide) is reduced by 1.46 times, and the content of Alpha-CEHC is increased by 1.31 times; accordingly, the controller is judged to be in a moderate fatigue state when the controller goes off duty from a white duty.
TABLE 1 biomarkers related to characteristic fatigue degree screened from urine samples of civil aviation air traffic controllers
Example two
1. Volunteer recruitment and sample collection
Volunteer recruitment: 25 volunteers of civil aviation air traffic controllers are recruited in an airport in a country, and the group entering conditions are as follows: healthy body, male, no medicine, age 20-35 years old; sample collection was performed in 2016 in autumn of 10 months.
Collecting a urine sample: the urine sample is collected once before the volunteer team of the control member goes on duty in 8 hours of work in white duty and is used as a non-fatigue sample, and the urine sample is collected once before the volunteer team of the control member goes off duty in white duty and is used as a fatigue sample. And collecting urine by using a sterile urine cup, and subpackaging the urine in a sterile tube for storage. The urine sample is subpackaged in a sterile centrifuge tube and stored at-80 ℃.
2. Pretreatment of urine samples
The urine sample pretreatment step comprises: prior to assay, urine samples stored at-80 ℃ were thawed at 0-4 ℃, centrifuged at 12000rpm for 5 minutes at 4 ℃, and 100. mu.L of the supernatant was diluted with 100. mu.L of water.
3. Analyzing and detecting urine sample by adopting liquid chromatogram-time-of-flight mass spectrometry
The urine sample is detected by using polar chromatographic column HILIC and weak polar chromatographic column C18And the non-polar chromatographic column PFPP is used for separating and detecting polar, weak-polar and non-polar components in the urine respectively.
The liquid phase condition of polar HILIC chromatographic column is UPLC BEH Amide HILIC column (2.1mm × 100mm,1.7 μm), and the mobile phase is A phase composed of 95% acetonitrile and 5% formic acid containing 0.1%The phase B composition is an aqueous solution containing 0.1 percent of formic acid; the column temperature was 40 ℃, the sample size was 2.0. mu.L, and the mobile phase flow rate was 0.3 mL/min. The mass spectrum conditions are as follows: positive ion ionization mode detection, mass range was set to 50-1200m/z full scan mode. The optimum capillary voltage for the electrospray ionizer was 3000V, and Cone voltage was 30V. The drying gas was nitrogen, the desolvation flow rate was 800L/h, and the cone gas flow rate was 30L/h. The desolvation temperature was 400 ℃ and the ion source temperature was 100 ℃. Leuconeeenkephalin (leucine enkephalin) with the concentration of 0.2ng/mL is used as a mass number calibration internal standard, and the mass of a calibration ion is 556.2771Da ([ M + H ]]+)。
Using weak polarity C18The liquid phase conditions of the chromatographic column are as follows: UPLC CSH C18column (2.1mm × 100mm,1.7 μ M), mobile phase A phase composition is aqueous solution containing 0.1% formic acid, phase B composition is acetonitrile, column temperature is 40 ℃, sample introduction amount is 2.0 μ L, mobile phase flow rate is 0.3mL/min, mass spectrum conditions are that two ionization modes of positive ions and negative ions are detected, mass range is set to be 50-1200M/z full scan mode, optimum capillary voltage of an electrospray ionization device is 3000V (positive ions) or 2200V (negative ions), Cone voltage is 30V, dry gas is nitrogen, desolvation flow rate is 800L/H, Cone air flow rate is 30L/H, desolvation temperature is 400 ℃, ion source temperature is 100 ℃, leucinenphaken (leucine enkephalin) with concentration of 0.2ng/mL is used as internal calibration standard for mass number calibration, and calibration ion mass is 556.2771Da ([ M + H ] (M + H + E]+) Or 554.2615Da ([ M-H)]-)。
The liquid phase conditions of the adopted nonpolar PFPP chromatographic column are that UPLC HSS PFPP column (2.1mm × 100mm,1.7 mu m), the mobile phase A phase composition is an aqueous solution containing 0.1% formic acid, the phase B composition is methanol, the column temperature is 40 ℃, the sample injection amount is 2.0 mu L, the flow rate of the mobile phase is 0.3mL/min, the mass spectrum conditions are that two ionization modes of positive ions and negative ions are detected, the mass range is set to be 50-1200m/z full scanning mode, the optimal capillary tube voltage of the electrospray ionizer is 3000V (positive ions) or 2200V (negative ions), the Cone voltage is 30V, the dry gas is nitrogen, the desolvation flow rate is 800L/h, the Cone gas flow rate is 30L/h, the desolvation temperature is 400 ℃, the ion source temperature is 100 ℃, and leucoxene with the ion source concentration of 0.2ng/mL is adoptedkephalin (leucine enkephalin) as internal standard for mass number calibration, and mass of calibration ion is 556.2771Da ([ M + H ]]+) Or 554.2615Da ([ M-H)]-)。
Each urine sample is respectively detected by 3 chromatographic columns for polar, weakly polar and nonpolar components, wherein the polar component is only detected in a positive ion mode, and the weakly polar and nonpolar components are detected in a positive ion mode and a negative ion mode, namely each urine sample is detected for 5 times, so that all metabolic compounds in the urine can be detected as far as possible. A blank is inserted into every 10 samples in the detection to prevent cross contamination, and a quality control sample is inserted for quality control. The temperature of the sample chamber was maintained at 4 ℃ during the analytical test.
4. Data processing
And analyzing detection data obtained by detecting the sample by adopting a liquid chromatography/mass spectrometer, and respectively performing statistical analysis processing including data alignment and peak extraction by adopting a metabonomics data processing professional software Progenetics QI. And (3) pairing and grouping the detection result of the urine sample after work and the detection result of the urine sample before work, processing the detection data in the step (3) by adopting an orthogonal partial least squares method (OPLS-DA), and primarily screening the characteristic biomarker by referring to the retention characteristics of the chromatogram and the mass spectrum characteristics according to the condition that P is less than or equal to 0.05, CV is less than or equal to 30%, VIP is greater than 1.0 and the maximum change multiple is greater than or equal to 1.2 as a screening threshold.
Analyzing and screening 4 compounds as characteristic biomarkers in data of 7903 compounds of polar components in urine by using a polar HILIC column positive ion mode; in the presence of weak polarity C18In the data of 10463 compounds of weak polar components in the urine detected by the positive ion mode of the column, 5 compounds are analyzed and screened as characteristic biomarkers; using weak polarity C18In the data of 6014 compounds of weakly polar components in urine detected in a column negative ion mode, 1 compound is analyzed and screened as a characteristic biomarker; analyzing and screening 4 compounds as characteristic biomarkers in the data of 9739 compounds for detecting the nonpolar component in the urine by using the nonpolar PFPP column positive ion mode; 5996 compounds for detecting nonpolar components in urine by utilizing nonpolar PFPP column negative ion modeThe data in (2) were not analyzed to screen for a qualified characteristic biomarker. Specific results are shown in table 2.
5. Comparing the characteristic markers and judging the fatigue degree of the team
Examining the characteristic biomarkers screened in table 2, the biomarkers related to the fatigue degree of the team of civil aviation air traffic controllers were included: urocanic acid (Urocanic acid), acetylcytosine (N4-acylcytidine), 5-hydroxytryptophan (5-Hydroxy-L-tryptophan), dimethylguanosine (N2, N2-Dimehtylguanosine), acetanilide (N-Acetylarylamine), and Alpha-CEHC. Further checking the change multiple of each biomarker, the content of urocanic acid (Urocacic) is reduced by 1.44 times, the content of acetylcytosine (N4-Acetylcytidine) is reduced by 1.36 times, the content of 5-hydroxytryptophan (5-Hydroxy-L-tryptophan) is reduced by 1.62 times, the content of dimethylguanosine (N2, N2-Dimehtylguanosine) is reduced by 1.50 times, the content of acetanilide (N-acetylarylamide) is reduced by 1.78 times and the content of Alpha-CEHC is increased by 1.33 times; accordingly, the controller is judged to be in a moderate fatigue state when the controller goes off duty from a white duty.
TABLE 2 biomarkers related to characteristic fatigue degree screened from urine samples of civil aviation air traffic controllers
Comparative example
1. Volunteer recruitment and sample collection
Volunteer recruitment: 23 volunteers of administrative staff were recruited at an airport in a country, and the group entry conditions were as follows: healthy body, male, no medicine, age 20-35 years old; sample collection was performed in 2016 in autumn of 10 months.
Collecting a urine sample: the urine sample is collected once before 8 hours of work of the executive logistics worker on duty as a non-fatigue sample, and the urine sample is collected once after the executive logistics worker is off duty. And collecting urine by using a sterile urine cup, and subpackaging the urine in a sterile tube for storage. The urine sample is subpackaged in a sterile centrifuge tube and stored at-80 ℃.
2. Pretreatment of urine samples
The urine sample pretreatment step comprises: prior to assay, urine samples stored at-80 ℃ were thawed at 0-4 ℃, centrifuged at 12000rpm for 5 minutes at 4 ℃, and 100. mu.L of the supernatant was diluted with 100. mu.L of water.
3. Analyzing and detecting urine sample by adopting liquid chromatogram-time-of-flight mass spectrometry
The urine sample is detected by using polar chromatographic column HILIC and weak polar chromatographic column C18And the non-polar chromatographic column PFPP is used for separating and detecting polar, weak-polar and non-polar components in the urine respectively.
The liquid phase conditions of a polar HILIC chromatographic column are UPLC BEH Amide HILIC column (2.1mm × 100mm,1.7 mu M), the mobile phase is a water solution with the composition of phase A being 95% acetonitrile and 5% formic acid, the phase B being a water solution with the composition of 0.1% formic acid, the column temperature is 40 ℃, the sample injection amount is 2.0 mu L, the flow rate of the mobile phase is 0.3mL/min, the mass spectrum conditions are that the detection is in a positive ion ionization mode, the mass range is 50-1200M/z full scan mode, the optimal capillary voltage of an electrospray ionization device is 3000V, the Cone voltage is 30V, the dry gas is nitrogen, the desolvation flow rate is 800L/H, the Cone gas flow rate is 30L/H, the desolvation temperature is 400 ℃, the ion source temperature is 100 ℃, leucinenkephalin a leucinenkephalin (leucine enkephalin) with the concentration of 0.2ng/mL is used as the mass internal standard, and the ion mass is 556.2771Da M + M]+)。
Using weak polarity C18The liquid phase conditions of the chromatographic column are as follows: UPLC CSH C18column (2.1mm × 100mm,1.7 μm), mobile phase A phase composition is water solution containing 0.1% formic acid, phase B composition is acetonitrile, column temperature is 40 deg.C, sample volume is 2.0 μ L, mobile phase flow rate is 0.3mL/min, mass spectrum conditions are that two ionization modes of positive ion and negative ion are detected, mass range is set to 50-1200m/z full scan mode, optimum capillary voltage of electrospray ionization device is 3000V (positive ion) or 2200V (negative ion), Cone voltage is 30V, dry gas is nitrogen, desolvation flow rate is 800L/h, Cone air flow rate is 30L/h, desolvation temperature is 400 deg.C, ion source temperature is 100 deg.C, leucinenphalin (bright ammonia) with concentration of 0.2ng/mL is usedEnkephalin acid) as internal standard for mass number calibration, and the mass of calibration ion is 556.2771Da ([ M + H ]]+) Or 554.2615Da ([ M-H)]-)。
The liquid phase conditions of a nonpolar PFPP chromatographic column are UPLC HSS PFPP column (2.1mm × 100mm,1.7 mu M), the mobile phase is phase A which is an aqueous solution containing 0.1% formic acid, phase B which is methanol, the column temperature is 40 ℃, the sample volume is 2.0 mu L, the mobile phase flow rate is 0.3mL/min, the mass spectrum conditions are that two ionization modes of positive ions and negative ions are detected, the mass range is set to be 50-1200M/z full scanning mode, the optimal capillary voltage of an electrospray ionizer is 3000V (positive ions) or 2200V (negative ions), the Cone voltage is 30V, the drying gas is nitrogen, the desolvation flow rate is 800L/H, the Cone gas flow rate is 30L/H, the desolvation temperature is 400 ℃, the ion source concentration is 0.2ng/mL, leucineenkephalin (leucine enkephalin) is used as a mass number calibration internal standard, and the calibration ion mass is 556.2771 ([ M + H ] + Da H [ + H [ ([ M + ]]+) Or 554.2615Da ([ M-H)]-)。
Each urine sample is respectively detected by 3 chromatographic columns for polar, weakly polar and nonpolar components, wherein the polar component is only detected in a positive ion mode, and the weakly polar and nonpolar components are detected in a positive ion mode and a negative ion mode, namely each urine sample is detected for 5 times, so that all metabolic compounds in the urine can be detected as far as possible. A blank is inserted into every 10 samples in the detection to prevent cross contamination, and a quality control sample is inserted for quality control. The temperature of the sample chamber was maintained at 4 ℃ during the analytical test.
4. Data processing
And analyzing detection data obtained by detecting the sample by adopting a liquid chromatography/mass spectrometer, and respectively performing statistical analysis processing including data alignment and peak extraction by adopting a metabonomics data processing professional software Progenetics QI. And (3) pairing and grouping the detection result of the urine sample after work and the detection result of the urine sample before work, processing the detection data in the step (3) by adopting an orthogonal partial least squares method (OPLS-DA), and primarily screening the characteristic biomarker by referring to the retention characteristics of the chromatogram and the mass spectrum characteristics according to the condition that P is less than or equal to 0.05, CV is less than or equal to 30%, VIP is greater than 1.0 and the maximum change multiple is greater than or equal to 1.2 as a screening threshold.
Analyzing and screening 10 compounds as characteristic biomarkers from 8315 compounds of polar components in urine by using a polar HILIC column positive ion mode; in the presence of weak polarity C18In the data of 10645 compounds of weak polar components in the urine detected by the positive ion mode of the column, 17 compounds are analyzed and screened as characteristic biomarkers; using weak polarity C18In the data of 6966 compounds of weak polar components in the urine detected by the column negative ion mode, 7 compounds are analyzed and screened as characteristic biomarkers; analyzing and screening 10 compounds as characteristic biomarkers in the data of 10318 compounds of nonpolar components in urine by using a nonpolar PFPP column positive ion mode; in the data of 6571 compounds of nonpolar components in urine detected by using a nonpolar PFPP column negative ion mode, a characteristic biomarker meeting the condition cannot be analyzed and screened. Specific results are shown in table 3.
5. Comparing the characteristic markers and judging the fatigue degree of administrative and logistic personnel
Examining the characteristic biomarkers screened in table 3, only the biomarkers related to the fatigue degree of the civil aviation air traffic controller team were included: urocanic acid (Urocanic acid), acetylcytosine (N4-Acetylcytidine), and 5-hydroxytryptophan (5-Hydroxy-L-tryptophan). Further checking the change multiple of each biomarker, the content of urocanic acid (Urocacic) is reduced by 1.59 times, the content of acetylcytosine (N4-Acetylcytidine) is reduced by 1.37 times, and the content of 5-hydroxytryptophan (5-Hydroxy-L-tryptophan) is reduced by 1.43 times; judging that the administrative and logistics personnel are not moderately tired according to the judgment result; and accordingly, the type and content change of the characteristic biomarker when administrative logistics staff leave work for 8 hours from work is judged to be greatly different from the type and content change of the characteristic biomarker before leaving work after leaving work of 8 hours from work of a white duty of a civil aviation air traffic controller (8-hour from work of the white duty).
TABLE 3 biomarker related to characteristic fatigue degree screened from urine samples of administrative staff of control group
It is to be noted that, unless otherwise specified, technical or scientific terms used herein shall have the ordinary meaning as understood by those skilled in the art to which the invention pertains. Unless specifically stated otherwise, the relative steps, numerical expressions, and values of the components and steps set forth in these embodiments do not limit the scope of the present invention. In all examples shown and described herein, unless otherwise specified, any particular value should be construed as merely illustrative, and not restrictive, and thus other examples of example embodiments may have different values.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention, and all of the technical solutions are covered in the protective scope of the present invention.
Claims (7)
1. A method for detecting the mild and moderate fatigue degrees of a civil aviation air traffic controller team is characterized by comprising the following steps:
s1: collecting urine samples of a team of civil aviation air traffic controllers, and then preprocessing the urine samples; wherein the urine samples comprise urine samples before work and urine samples after work;
s2: by liquid chromatography-while-flyingPerforming separation detection on the pretreated samples by using a matrix spectroscopy; wherein the liquid chromatography adopts polar HILIC chromatographic column and low-polarity C18Separating and detecting the chromatographic column and the nonpolar PFPP chromatographic column;
s3: matching and comparing the detection result of the urine sample after the team works with the detection result of the urine sample before the team works, and screening out a characteristic biomarker;
s4: comparing the screened characteristic biomarkers with biomarkers related to fatigue degree of a team of civil aviation air traffic controllers, and judging the fatigue degree of the civil aviation air traffic controllers; biomarkers associated with the degree of mild and moderate fatigue of a civil aviation air traffic controller team include, but are not limited to: urocanic acid (C)6H6N2O2HMDB00301), acetylcytosine (N4-Acetylcytidine, C11H15N3O6HMDB 05923), 5-hydroxytryptophan (5-Hydroxy-L-tryptophan, C11H12N2O3HMDB 00472), Dimethylguanosine (N2, N2-Dimehtylguanosine, C12H17N5O5HMDB 04284), acetanilide (N-Acetylarylamine, C)8H9NO, HMDB 01250) and Alpha-CEHC (C)16H22O4，HMDB 01518)；
In the characteristic biomarkers screened in the step S3, urocanic acid, acetylcytosine, 5-hydroxytryptophan, dimethylguanosine, acetanilide and Alpha-CEHC are searched, and according to the types and the variation ranges of the characteristic markers, the condition that the civil aviation air traffic controller team has light or moderate fatigue can be judged;
after the team works, the detection result of the urine sample which works for 2-3 hours in the day is selected and compared with the detection result of the urine sample before the team works in a matching way, and if urocanic acid is detected and the content of the urocanic acid is up to or more than 1.4 times, the mild fatigue of the team of the civil aviation air traffic controller can be judged;
after the team works, the detection result of the urine sample which works for 3-5 hours in the day is selected and compared with the detection result of the urine sample before the team works in a matching way, and if urocanic acid, acetylcytosine and 5-hydroxytryptophan are detected in the screened characteristic markers, and the content of the urocanic acid, the acetylcytosine and the 5-hydroxytryptophan is respectively reduced to be more than 1.4 times, 1.3 times and more than 1.4 times, the moderate fatigue of the team of the civil aviation air traffic controller can be judged;
and (3) selecting a detection result of the urine sample which works for 5-8 hours in the daytime after the team works, matching and comparing the detection result with a detection result of the urine sample before the team works, and judging that the team of the civil aviation air traffic controller has moderate fatigue if urocanic acid, acetylcytosine, 5-hydroxytryptophan, dimethylguanosine, acetanilide and Alpha-CEHC are detected in the screened characteristic markers, and the content of the first five markers is respectively reduced to be more than 1.4, 1.3, 1.4 and 1.4 times and the content of the Alpha-CEHC is increased to be more than 1.3 times.
2. The method for detecting the mild and moderate fatigue degrees of the civil aviation air traffic controller team according to claim 1, characterized in that:
in step S1, the urine sample pretreatment includes: melting the urine sample stored at-80 ℃ at 0-4 ℃, centrifuging the urine sample at 4 ℃ and 12000rpm for 5min, and collecting the supernatant after centrifugation; 100 μ L of the supernatant was taken and then diluted with 100 μ L of water.
3. The method for detecting the mild and moderate fatigue degrees of the civil aviation air traffic controller team according to claim 1, characterized in that:
in the step S2, the temperature of the sample chamber is kept between 0 and 4 ℃ in the separation detection process; when the liquid chromatography adopts a polar HILIC chromatographic column to separate and detect polar components, the mass spectrometry adopts a positive ion ionization detection mode; the liquid chromatography adopts weak polarity C18When the chromatographic column is used for separating and detecting weak polar and nonpolar components, the mass spectrometry adopts two ionization modes of positive ions and negative ions; the liquid chromatography adopts nonpolar PFPP chromatographic column to separateWhen weak polar and non-polar components are detected, the mass spectrometry adopts two ionization modes of positive ions and negative ions.
4. The method for detecting the mild and moderate fatigue of the civil aviation air traffic controller team according to claim 3, characterized in that:
in step S2, when the liquid chromatography adopts a polar HILIC chromatographic column to separate and detect polar components, the mass spectrometry adopts a positive ion detection ionization mode, and the detection conditions are as follows: the mass range is set to 50-1200m/z full scan mode; the voltage of a capillary tube of the electrospray ionizer is 3000V, and the Cone voltage is 30V; the drying gas is nitrogen, the desolvation flow rate is 800L/h, and the cone gas flow rate is 30L/h; the desolvation temperature is 400 ℃, and the ion source temperature is 100 ℃; leucine enkephalin with the concentration of 0.2ng/mL is used as a mass number calibration internal standard, and the mass of a calibration ion is 556.2771Da [ M + H ]]+The liquid phase conditions are that UPLC BEH AmideHILIC column 2.1mm × 100mm,1.7 mu m, and the mobile phase comprises phase A and phase B, wherein the phase A comprises acetonitrile with volume fraction of 95% and aqueous solution with volume fraction of 5% containing 0.1% formic acid, the phase B comprises aqueous solution containing 0.1% formic acid, the column temperature is 40 ℃, the sample injection amount is 2.0 mu L, and the flow rate of the mobile phase is 0.3 mL/min.
5. The method for detecting the mild and moderate fatigue of the civil aviation air traffic controller team according to claim 3, characterized in that:
in step S2, the liquid chromatography is performed by using weak polarity C18When the chromatographic column is used for separating and detecting weak polar and nonpolar components, the mass spectrometry adopts two ionization modes of positive ions and negative ions, and the detection conditions are as follows: the mass range is set to 50-1200m/z full scan mode; the voltage of a capillary tube of the electrospray ionizer is 3000V positive ions or 2200V negative ions, and the cone voltage is 30V; the drying gas is nitrogen, the desolvation flow rate is 800L/h, and the cone gas flow rate is 30L/h; the desolvation temperature is 400 ℃, and the ion source temperature is 100 ℃; leucine enkephalin with the concentration of 0.2ng/mL is used as a mass number calibration internal standard, and the mass of a calibration ion is 556.2771Da [ M + H ]]+Or 554.2615Da [ M-H [ ]]-(ii) a The liquid phase conditions were: UPLC CSH C18column 2.1mm × 100mm,1.7 μm, mobile phase A as aqueous solution containing 0.1% formic acid, phase B as acetonitrile, column temperature 40 deg.C, sample size 2.0 μ L, and mobile phase flow rate 0.3 mL/min.
6. The method for detecting the mild and moderate fatigue of the civil aviation air traffic controller team according to claim 3, characterized in that:
in step S2, when the liquid chromatography employs a nonpolar PFPP chromatographic column to separate and detect weak polar and nonpolar components, the mass spectrometry employs two ionization modes, namely positive ions and negative ions, and the detection conditions are as follows: the mass range is set to 50-1200m/z full scan mode; the voltage of a capillary tube of the electrospray ionizer is 3000V positive ions or 2200V negative ions, and the cone voltage is 30V; the drying gas is nitrogen, the desolvation flow rate is 800L/h, and the cone gas flow rate is 30L/h; the desolvation temperature is 400 ℃, and the ion source temperature is 100 ℃; leucine enkephalin with the concentration of 0.2ng/mL is used as a mass number calibration internal standard, and the mass of a calibration ion is 556.2771Da [ M + H ]]+Or 554.2615Da [ M-H [ ]]-The liquid phase conditions are UPLC HSS PFPP column 2.1mm × 100mm,1.7 μm, mobile phase A is aqueous solution containing 0.1% formic acid, phase B is methanol, column temperature is 40 deg.C, sample injection amount is 2.0 μ L, and mobile phase flow rate is 0.3 mL/min.
7. The method for detecting the mild and moderate fatigue of the civil aviation air traffic controller team according to claim 3, characterized in that:
step S3 includes: carrying out statistical analysis processing on detection data obtained by adopting liquid chromatography-time-of-flight mass spectrometry by adopting metabonomics data processing software Progenetics QI;
the analysis processing method comprises data alignment and peak extraction; the method also comprises the steps of carrying out pairing grouping on the detection result of the urine sample of the civil aviation air traffic controller team after work and the detection result of the urine sample of the civil aviation air traffic controller team before work, then carrying out processing analysis by adopting a partial least square method OPLS-DA, and screening out the characteristic biomarker by taking P less than or equal to 0.05, CV less than or equal to 30%, VIP greater than 1.0 and maximum change multiple more than or equal to 1.2 as screening thresholds.
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