CN107607641A - Air traffic control person teams and groups are slight and the detection method of moderate degree of fatigue - Google Patents
Air traffic control person teams and groups are slight and the detection method of moderate degree of fatigue Download PDFInfo
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Abstract
The present invention relates to a kind of air traffic control person teams and groups are slight and the detection method of moderate degree of fatigue, comprise the following steps：S1：The urine specimen of air traffic control person teams and groups is gathered, then urine specimen is pre-processed；Wherein, urine specimen includes the urine specimen before working work and the urine specimen after work；S2：Separation detection is carried out using liquid chromatogram time-of-flight mass spectrometry (TOFMS) respectively to pretreated sample；S3：The testing result of urine specimen after teams and groups are worked and the urine specimen before working work carries out pairing and compared, and filters out characteristic biological mark；S4：Compared with by the characteristic biological mark of the screening biomarker related with air traffic control person's degree of fatigue, the degree of fatigue of air traffic control person is judged.Air traffic control person teams and groups provided by the invention are slight and the detection method of moderate degree of fatigue, easy to operate, with a high credibility.
Description
Technical field
The present invention relates to technical field of biological, and in particular to a kind of air traffic control person teams and groups are slightly with
Spend the detection method of degree of fatigue.
Background technology
The history of human research's fatigue can trace back to the World War I before a century earliest, in Great War and postwar Britain
Industrial fatigue research committee (renames as The after The Industrial Fatigue Research Board
Industrial Health Research Board) carry out one on fatigue in industrial production particularly military material production
The extensive ergonomics research of problem.The research report has a far reaching influence, or even many results are into the common recognition of industrial quarters.For example,
It the report recommends and shorten long working time, improve alleviation employee's fatigue such as working environment, adjustment break tour system, improve work life
Yield.These suggest design and the break tour systems such as the illumination to our working time settings now, workshop and working environment
System optimization still has directive function.The index for being used to detect fatigue in this part of report is exactly labor productivity.
In World War II, aircraft industry particularly military aviation industry is quickly grown.It is found that pilot exists
During button aircraft execution task, it may appear that driving ability declines the situation of even disability suddenly, it has been proposed that to operate achievement
Effect declines to characterize fatigue.After the war, develop with military aviation and Civil Aviation Industry, long-distance flight and round the clock flight become
It is more and more common, it has been found that fatigue problem more complicates, and fatigue can add up, and forms accumulation fatigue.This period, still
Based on ergonomics technique study fatigue.
The continuous improvement paid close attention to people social safety, the research for fatigue problem are also expanded by ergonomics research
To subjects such as psychology, Neuscience, hypnosphy, Physiology and biochemistries.The method that each subject establishes corresponding detection fatigue, such as the heart
Stamford Sleepiness Scale of the Neo-Confucianism based on subjective feeling, Visual analogue scale etc., the also prestige based on cognitive ability detection fatigue
This Kang Xing card sorting test (WSCT), Tower of London test (TOL) etc..Subjective scale is vulnerable to mood and other factors interference,
Parameter index setting is relatively difficult in recognition tests, " ceiling " or " floor " effect often occurs, causes these scales or soft
The reliability and validity of part are not satisfactory enough；And for example nerve and sleep science are established based on electroencephalogram change detection fatigue method,
It is proposed occur fatigue when delta and theta ripples significantly increase, because Brain Diagnostic Apparatus is complicated, it is difficult to applied to Site Detection；For another example
Occur yawning during based on human-body fatigue, the dynamic change of eye such as blink, catacleisis physiological reaction, attempt to establish accordingly detection side
Method.So far, people are understood tired biochemical mechanism seldom, and the explaination of tired biochemical mechanism will be helpful to exploitation based on biochemistry
Index it is objective, stably, quick detection fatigue method.
It is a kind of method that people have attempted to using biochemical indicator detection fatigue, such as detects some hormones, peptide, studies
It was found that cytidine (Cytidine) and adrenal cortex class ester alcohol (Corticoid) concentration are relevant with operating pressure；Also examine
Survey the material relevant with sleep, biological rhythm such as glycopeptide (Glycopeptides)；But Borbely etc. point out these compounds with
Fatigue has the evidence of the data deficiency uniformity of correlation.In a word, although people are from operating pressure, sleep, biological rhythm etc.
Multi-angle has attempted multiple compounds metabolism and tired correlation research, but does not find any body metabolism compound yet
There is correlation with fatigue.
Metabolism group is the new branch of science of metabolic characteristics in a system research body, mainly using nuclear magnetic resonance, gas phase
Chromatography-mass spectroscopy, liquid chromatography-mass spectrography etc. test and analyze means combination Chemical Measurement, systematically explore, find and find human body
The characteristics index of its body fluid metabolic is biomarker when being influenceed by disease, medicine etc., in medical diagnosis on disease, medicament research and development etc.
Field obtains extensive use.For example, Mapstone of Georgetown University of the U.S. etc. is using LC Mass (liquid matter point
Analysis) metabolism group method, examination goes out 10 kinds of characteristic biological marks, can be in the disease for detecting alzheimer's disease
2-3 is diagnosed before clinical symptoms occurs in disease, and accuracy rate can reach more than 90%；And for example, Naviaux etc. is using liquid matter point
Technique study chronic fatigue syndrome metabolic characteristics is analysed, it is found that male patient has 8 kinds of metabolins, 13 kinds of metabolism of female patient
Thing occurs significantly to sexually revise, Subject characteristics' sex work curve (Receiver operator characteristic curve,
ROC) analysis result shows that accuracy rate of diagnosis reaches 94% (male) and 96% (women)；Professor Xu Guowang etc. is using the analysis of liquid matter
Metabolism group research is carried out to depression and fatigue rat model blood and urine, the results showed that two groups of rats are with compareing
Group rat is compared, and anomalous variation occurs in metabolism.Armstrong etc. samples two dimensional NMR energy disperse spectroscopy to chronic fatigue syndrome
Patients serum has carried out metabolism group research, the results showed that case group serum GLN and ornithine content are substantially less than pair
According to group.In above-mentioned document report, packet comparison is carried out using case group and control group for various diseases, but because fatigue is
A kind of physiological change that can recover, existing literature method are not suitable for the screening of degree of fatigue associated biomarkers.
Air traffic control person, the air traffic air route that takes charge are boarded a plane safe operation, are generally just referred to
Aircraft is waved, manages all airborne vehicles in whole spatial domain as a whole with reference to information such as radars.Controller often commands simultaneously at work
The operation of multi-aircraft, it is necessary to energy concentrate, the ability such as accurate judgement and emergency processing, therefore be very easy to fatigue, civil aviation authority
Relevant regulations clearly every controller is no more than 2 hours in seat commander's aircraft maximum duration.It is main that fatigue is produced in its work
It is caused by mental labour.Chen etc. utilizes liquid chromatography-mass spectrometry in the literature, to the urine of air traffic control person
3 kinds of tired associated biomarkers of screening sample, but the concept of degree of fatigue is not proposed, in addition do not disclose yet and to utilize fatigue
The method that associated biomarkers detect air traffic control person's degree of fatigue.The present inventor is established available for people early stage
The method of body fatigue associated biomarkers screening.On this basis, the present inventor has found by substantial amounts of experiment, degree of fatigue
It is related to air traffic control person's potential job safety executive capability, therefore degree of fatigue detection can give civil aviaton's air traffic pipe
Member's security risk management and control processed provides strong new technology and new tool.
The content of the invention
For in the prior art the defects of, the present invention establishes a kind of to be detected using sewage sludge method
Air traffic control person teams and groups are slight and the method for moderate degree of fatigue, available for pre-job check and evaluation whether can more than
Hilllock, detect in hilllock for finding tired risk and in time adjustment etc., for the detection of air traffic control person's degree of fatigue and prison
Survey, management and control fatigue risk, ensure that safety of civil aviation operation provides new technology and method.
To achieve the above object, technical scheme provided by the invention is：
The invention provides a kind of air traffic control person teams and groups slightly and moderate degree of fatigue detection method, wrap
Include following steps：S1：The urine specimen of air traffic control person is gathered, then urine specimen is pre-processed；Wherein,
Urine specimen includes the urine specimen before working work and the urine specimen after work；S2：Using liquid chromatogram-flight time matter
Spectrometry carries out separation detection respectively to pretreated sample；Wherein, liquid chromatogram can use polarity HILIC chromatographic columns, weak
Polarity C18One or more progress separation detections in chromatographic column and nonpolar PFPP chromatographic columns；S3：By the urine sample after work
The testing result of this testing result and the urine specimen before working work carries out pairing and compared, and screens characteristic biological mark
Thing；S4：By the characteristic biological mark of the screening biological marker related to air traffic control person teams and groups degree of fatigues
Thing is compared, and judges the degree of fatigue of air traffic control person teams and groups.
In step S3, the biomarker related to air traffic control person teams and groups degree of fatigues includes but unlimited
In：Urocanic acid (Urocanic acid, C6H6N2O2, HMDB 00301), acetylcytosine (N4-Acetylcytidine,
C11H15N3O6, HMDB 05923), 5-hydroxyryptophan (5-Hydroxy-L-tryptophan, C11H12N2O3, HMDB 00472),
Dimethylguanosine (N2, N2-Dimehtylguanosine, C12H17N5O5, HMDB 04284), antifebrin (N-
Acetylarylamine, C8H9NO, HMDB 01250) and Alpha-CEHC (C16H22O4, HMDB 01518).
Preferably, the testing result of the urine specimen after the work of air traffic control person teams and groups and civil aviaton's air traffic
The testing result paired comparisons of urine specimen before the working work of controller teams and groups, screen characteristic biological mark；With civil aviaton
In the related characteristic biological mark of air traffic controller teams and groups degree of fatigue：Search urocanic acid, acetylcytosine, 5- hydroxyls
Tryptophan, dimethylguanosine, antifebrin and Alpha-CEHC, the species occurred according to features described above mark and its change
Scope, it can determine whether that slight or moderate fatigue occur in air traffic control person teams and groups.
Preferably, preferably work by day the urine specimen after 2-3 hours after the work of air traffic control person teams and groups
The testing result paired comparisons of urine specimen before testing result and the working work of air traffic control person teams and groups, are filtered out
Markers characteristic in, if detection urocanic acid, and the up-regulation of its content meets or exceeds 1.4 times, you can judges that civil aviaton is empty
There is slight fatigue in middle traffic controller teams and groups.
Preferably, preferably work by day the urine specimen after 3-5 hours after the work of air traffic control person teams and groups
The testing result paired comparisons of urine specimen before testing result and the working work of air traffic control person teams and groups, are filtered out
Markers characteristic in, if detection urocanic acid, acetylcytosine, 5-hydroxyryptophan, and their contents decline reach respectively
Arrive and more than more than 1.4,1.3,1.4 times, you can judge that moderate fatigue occur in air traffic control person teams and groups.
Preferably, preferably work by day the urine specimen after 5-8 hours after the work of air traffic control person teams and groups
The testing result paired comparisons of urine specimen before testing result and the working work of air traffic control person teams and groups, are filtered out
Markers characteristic in, if detection urocanic acid, acetylcytosine, 5-hydroxyryptophan, dimethylguanosine, antifebrin and
Alpha-CEHC, and first five kind content decline respectively reaches and more than more than 1.4,1.3,1.4,1.4,1.4 times and Alpha-
The rise of CEHC contents reaches and more than more than 1.3 times, you can judges that moderate fatigue occur in air traffic control person teams and groups.
In step S1, urine specimen pretreatment includes：- 80 DEG C of urine specimens will be stored at 0-4 DEG C to melt, then 4
DEG C, 12000rpm centrifugation urine specimen 5min, supernatant is collected after centrifugation；100 μ L of supernatant liquid are taken, then with 100 μ L water
Dilution.
In step S2, sample room temperature is kept for 0-4 DEG C during separation detection；Liquid chromatogram uses polarity HILIC colors
When composing post separation detection polar compound, mass spectrography is using detection cation ionization mode；Liquid chromatogram uses low pole C18Color
When composing post separation detection low pole and non-polar component, mass spectrography uses two kinds of ionization modes of cation and anion；Liquid phase
When chromatogram uses nonpolar PFPP chromatographic columns separation detection low pole and non-polar component, mass spectrography uses cation and anion
Two kinds of ionization modes.
Preferably, in step S2, when liquid chromatogram uses polarity HILIC chromatographic column separation detection polar compounds, mass spectrography
Using detection cation ionization mode, testing conditions are：Mass range is arranged to 50-1200m/z full scan patterns；Electron spray
The optimal capillary voltage of ionizer is 3000V, and cone voltages are 30V；It is nitrogen to dry gas, and desolvation flow velocity is 800L/
H, cone gas velocity are 30L/h；Desolvation temperature is 400 DEG C, and ion source temperature is 100 DEG C；Concentration is used as 0.2ng/mL
Leucineenkephalin (leucine enkephalin) as mass number calibration internal standard, calibration mass of ion is 556.2771Da
([M+H]+)；Liquid-phase condition is：UPLC BEH Amide HILIC column(2.1mm×100mm,1.7μm)；Mobile phase bag
Include A phases and B phases, A phase compositions are the acetonitrile of volume fraction 95% and volume fraction is 5% aqueous solution containing 0.1% formic acid, B phases
Form as the aqueous solution containing 0.1% formic acid；Column temperature is 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.
Preferably, in step S2, liquid chromatogram uses low pole C18Chromatographic column separation detection low pole and non-polar component
When, mass spectrography uses two kinds of ionization modes of cation and anion, and testing conditions are：Mass range is arranged to 50-1200m/z
Full scan pattern；The optimal capillary voltage of electro-spray ionization device is 3000V (cation) or 2200V (anion), cone electricity
Press as 30V；It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are 30L/h；Desolvation temperature is 400
DEG C, ion source temperature is 100 DEG C；Use leucineenkephalin (leucine enkephalin) conduct of concentration for 0.2ng/mL
Mass number calibrates internal standard, and calibration mass of ion is 556.2771Da ([M+H]+) or 554.2615Da ([M-H]-)；Liquid-phase condition
For：UPLC CSH C18column(2.1mm×100mm,1.7μm)；Mobile phase A phase composition is to contain the water-soluble of 0.1% formic acid
Liquid, B phase compositions are acetonitrile；Column temperature is 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.
Preferably, in step S2, liquid chromatogram uses nonpolar PFPP chromatographic columns separation detection low pole and nonpolar group
Timesharing, mass spectrography use two kinds of ionization modes of cation and anion, and testing conditions are：Mass range is arranged to 50-
1200m/z full scan patterns；The optimal capillary voltage of electro-spray ionization device is 3000V (cation) or 2200V (anion),
Cone voltages are 30V；It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are 30L/h；Desolvation temperature
Spend for 400 DEG C, ion source temperature is 100 DEG C；Use leucineenkephalin (leucine brain coffee of the concentration for 0.2ng/mL
Peptide) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M+H]+) or 554.2615Da ([M-H]-)；Liquid
Phase condition is：UPLC HSS PFPP column(2.1mm×100mm,1.7μm)；Mobile phase is that A phase compositions are to contain 0.1%
The aqueous solution of formic acid, B phase compositions are methanol；Column temperature is 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.
Preferably, step S3 is specifically included：The testing number that will be obtained using the detection of sewage sludge method
According to using data processing software, it is preferred to use metabolism group data processing related software carries out statistical analysis processing, further excellent
Choosing carries out statistical analysis processing using metabolism group data processing arts software Pro genesis QI；Analysis and processing method includes
Alignment of data, peak extraction；Also include testing result and the people of the urine specimen after working air traffic control person teams and groups
The testing result of urine specimen before the working work of boat air traffic controller teams and groups carries out pairing packet, then using partially minimum
Square law OPLS-DA Treatment Analysis, with P≤0.05, CV≤30%, VIP>1.0th, maximum change multiple >=1.2 are as screening threshold
Value, filters out characteristic biological mark.
Technical scheme provided by the invention, there is following beneficial effect：(1) air traffic control person class of the present invention
Group is slight and the detection method of moderate degree of fatigue, and testing result is with a high credibility, and the present invention can use a variety of opposed polarities
Chromatographic column carries out repeated detection to same sample, on the one hand can avoid missing inspection, on the other hand can be same according to different chromatographic columns
When detect same composition result the detection method of chromatographic column is verified, avoid a certain chromatographic column from error occur, so as to carry
High accuracy；(2) air traffic control person, the operation of multi-aircraft often is commanded, it is necessary to energy collection simultaneously at work
In, the ability such as accurate judgement and emergency processing, therefore be very easy to fatigue；Present invention applicant tests discovery by substantial amounts of,
Degree of fatigue is related to air traffic control person's potential job safety executive capability；Slight and moderate fatigue journey provided by the invention
Detection method is spent, is used available for whether the pre-job check and evaluation of air traffic control person teams and groups can detect with duty, in hilllock
In the tired risk of discovery and adjustment etc. in time；It is that civil aviaton hands in the air for air traffic control person teams and groups workload assessment
The degree of fatigue detection of logical controller teams and groups and monitoring, management and control fatigue risk, ensure that safety of civil aviation operation provides new technology and Xin Fang
Method.
The additional aspect and advantage of the present invention will be set forth in part in the description, and will partly become from the following description
Obtain substantially, or recognized by the practice of the present invention.
Brief description of the drawings
Fig. 1 is the polarity HILIC post cations of air traffic control person's urine specimen in the embodiment of the present invention one
Mode detection typical case's total ion current figure；
Fig. 2 is the HILIC post positive ion detections of air traffic control person's urine specimen in the embodiment of the present invention one
Polar compound data principal component analysis shot chart；
Fig. 3 is the low pole C of air traffic control person's urine specimen in the embodiment of the present invention one18Post cation mould
Formula detects typical total ion current figure；
Fig. 4 is the C of air traffic control person's urine specimen in the embodiment of the present invention one18The weak pole of post positive ion detection
Property component data principal component analysis shot chart；
Fig. 5 is the low pole C of air traffic control person's urine specimen in the embodiment of the present invention one18Post anion mould
Formula detects typical total ion current figure；
Fig. 6 is the C of air traffic control person's urine specimen in the embodiment of the present invention one18The weak pole of post anionic textiles
Property component data principal component analysis shot chart；
Fig. 7 is the nonpolar PFPP posts cation of air traffic control person's urine specimen in the embodiment of the present invention one
Mode detection typical case's total ion current figure；
Fig. 8 is that the PFPP post positive ion detections of air traffic control person's urine specimen in the embodiment of the present invention one are non-
Polar compound data principal component analysis shot chart；
Fig. 9 is the nonpolar PFPP posts anion of air traffic control person's urine specimen in the embodiment of the present invention one
Mode detection typical case's total ion current figure；
Figure 10 is the PFPP post anionic textiles of air traffic control person's urine specimen in the embodiment of the present invention one
Non-polar component data principal component analysis shot chart.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation describes.The following examples are only intended to illustrate the technical solution of the present invention more clearly, therefore is intended only as example, without
It can be limited the scope of the invention with this.Experimental method in following embodiments, it is conventional side unless otherwise specified
Method.Test material used, is to be commercially available from regular shops unless otherwise specified in following embodiments.
Embodiment one
1st, volunteer recruiting and sample collection
Volunteer recruiting：The people of air traffic control person volunteer 20 is recruited in certain International airport at home, enters a group condition：
Health, male, no medication, age 20-35 year, sample collection were carried out winter in December, 2014.
The collection of urine specimen：Once urinated in the day shift work pre-job collection of controller volunteer teams and groups working processed in 8 hours
Liquid sample gathers a urine specimen as tired before leaving offices as non-tired sample after controller volunteer teams and groups day shift is laid off
Labor sample.Urine specimen is collected urine and is sub-packed in sterile tube and preserved using Sterilized urine cup.Urine specimen be sub-packed in it is sterile from
Heart pipe is stored in -80 DEG C.
2nd, the pretreatment of urine specimen
Urine specimen pre-treatment step is：- 80 DEG C of urine specimens will be stored in before analysis detection at 0-4 DEG C to melt, 4
DEG C, 12000rpm is centrifuged 5 minutes, then takes the μ L of supernatant 100 to add the dilution of 100 μ L water.
3rd, analysis detection is carried out to urine specimen using sewage sludge method
Polarity chromatographic column HILIC, low pole chromatographic column C is respectively adopted in the detection of urine specimen18With nonpolar chromatographic column
PFPP carries out separation detection respectively to the polarity in urine, low pole and non-polar component.
Use the liquid-phase conditions of polarity HILIC chromatographic columns for：UPLC BEH Amide HILIC column(2.1mm×
100mm, 1.7 μm), mobile phase is that A phase compositions are 95% acetonitrile and 5% aqueous solution containing 0.1% formic acid, B phase compositions be containing
There is the aqueous solution of 0.1% formic acid；Column temperature is 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：Just
Ion mode detection, mass range are arranged to 50-1200m/z full scan patterns.The optimal capillary of electro-spray ionization device
Tube voltage is 3000V, and cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are
30L/h.Desolvation temperature is 400 DEG C, and ion source temperature is 100 DEG C.Concentration is used as 0.2ng/mL's
For leucineenkephalin (leucine enkephalin) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M
+H]+)。
Using low pole C18The liquid-phase condition of chromatographic column is：UPLC CSH C18column(2.1mm×100mm,1.7μ
M), mobile phase A phase composition is the aqueous solution containing 0.1% formic acid, and B phase compositions are acetonitrile；Column temperature is 40 DEG C, the μ L of sample size 2.0,
Flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：The two kinds of ionization mode detections of cation and anion, mass range are set
For 50-1200m/z full scan patterns.The optimal capillary voltage of electro-spray ionization device is 3000V (cation) or 2200V is (negative
Ion), cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are 30L/h.Go molten
Agent temperature is 400 DEG C, and ion source temperature is 100 DEG C.Use leucineenkephalin (bright ammonia of the concentration for 0.2ng/mL
Sour enkephalins) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M+H]+) or 554.2615Da ([M-
H]-)。
Use the liquid-phase conditions of nonpolar PFPP chromatographic columns for：UPLC HSS PFPP column(2.1mm×100mm,
1.7 μm), mobile phase is that A phase compositions are the aqueous solution containing 0.1% formic acid, and B phase compositions are methanol；Column temperature is 40 DEG C, sample size
2.0 μ L, flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：The two kinds of ionization mode detections of cation and anion, quality model
Enclose and be arranged to 50-1200m/z full scan patterns.The optimal capillary voltage of electro-spray ionization device be 3000V (cation) or
2200V (anion), Cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are
30L/h.Desolvation temperature is 400 DEG C, and ion source temperature is 100 DEG C.Concentration is used as 0.2ng/mL's
For leucineenkephalin (leucine enkephalin) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M
+H]+) or 554.2615Da ([M-H]-)。
Each urine specimen detects polarity, low pole and non-polar component therein respectively using 3 kinds of chromatographic columns, wherein
Polar compound only detects positive ion mode, low pole and non-polar component detection cation and negative ion mode, i.e., each urine
Pattern detection 5 times, to detect metabolic compounds whole in urine as far as possible.Every 10 samples insert a sky in detection
It is white to prevent cross pollution, and insert a Quality Control sample and carry out quality control.Sample room temperature is kept for 4 DEG C in analyte detection process.
4th, data processing
The detection data of detection sample acquisition are analyzed using liquid chromatography/mass spectrometry combined instrument, metabolism group number is respectively adopted
Statistical analysis processing, including the extraction of alignment of data, peak are carried out according to processing professional software Progenesis QI.By the urine after work
The testing result of the testing result of liquid sample and the urine specimen before working work carries out pairing packet, using an orthogonal most young waiter in a wineshop or an inn partially
The detection data of multiplication (OPLS-DA) processing step 3, according to P≤0.05, CV≤30%, VIP>1.0th, maximum change multiple >=
1.2 as screening threshold value, and with reference to chromatogram retention characteristic and mass spectral characteristic, preliminary screening goes out characteristic biological mark.
In using polarity HILIC posts positive ion mode detection urine in the data of 9490 compounds of polar compound, point
Analysis filters out 5 kinds of compound being characterized property biomarkers；Utilizing low pole C18Weak pole in post positive ion mode detection urine
In the data of property 10809 compounds of component, Analysis and Screening goes out 12 kinds of compound being characterized property biomarkers；Utilize low pole
C18Post negative ion mode detection urine in 5247 compounds of weakly polar component data in, fail Analysis and Screening go out it is eligible
Characteristic biological mark；11414 changes of non-polar component in using nonpolar PFPP posts positive ion mode detection urine
In the data of compound, Analysis and Screening goes out 6 kinds of compound being characterized property biomarkers；Utilizing nonpolar PFPP posts anion mould
In formula detection urine in the data of 6176 compounds of non-polar component, Analysis and Screening goes out 2 kinds of being characterized property of compound biology marks
Will thing.Concrete outcome is as shown in table 1.
5th, markers characteristic compares, and judges teams and groups' degree of fatigue
The characteristic biological mark filtered out in inspection table 1, contain air traffic control person teams and groups degree of fatigue
Related biomarker：Urocanic acid (Urocanic acid), acetylcytosine (N4-Acetylcytidine), 5- hydroxyl color ammonia
Sour (5-Hydroxy-L-tryptophan), dimethylguanosine (N2, N2-Dimehtylguanosine), antifebrin (N-
) and Alpha-CEHC Acetylarylamine.Further check each biomarker change multiple, urocanic acid (Urocanic
Acid) content declines 1.41 times, the decline of acetylcytosine (N4-Acetylcytidine) content 1.39 times, 5-hydroxyryptophan (5-
Hydroxy-L-tryptophan) under 1.42 times of content decline, dimethylguanosine (N2, N2-Dimehtylguanosine) content
1.41 times of drop, antifebrin (N-Acetylarylamine) content decline 1.46 times and Alpha-CEHC contents rise 1.31 times；
Judge the controller teams and groups day shift to be laid off when coming off duty accordingly and be in moderate fatigue state.
The characteristic fatigue degree associated biomarkers filtered out in the air traffic control person's urine specimen of table 1
Embodiment two
1st, volunteer recruiting and sample collection
Volunteer recruiting：The people of air traffic control person volunteer 25 is recruited in certain International airport at home, enters a group condition：
Health, male, no medication, age 20-35 year；Sample collection was carried out autumn in October, 2016.
The collection of urine specimen：Once urinated in the day shift work pre-job collection of controller volunteer teams and groups working processed in 8 hours
Liquid sample gathers a urine specimen as tired before leaving offices as non-tired sample after controller volunteer teams and groups day shift is laid off
Labor sample.Collect urine using Sterilized urine cup and be sub-packed in sterile tube and preserve.Urine specimen is sub-packed in sterile centrifugation tube preservation
In -80 DEG C.
2nd, the pretreatment of urine specimen
Urine specimen pre-treatment step is：- 80 DEG C of urine specimens will be stored in before analysis detection at 0-4 DEG C to melt, 4
DEG C, 12000rpm is centrifuged 5 minutes, then takes the μ L of supernatant 100 to add the dilution of 100 μ L water.
3rd, analysis detection is carried out to urine specimen using sewage sludge method
Polarity chromatographic column HILIC, low pole chromatographic column C is respectively adopted in the detection of urine specimen18With nonpolar chromatographic column
PFPP carries out separation detection respectively to the polarity in urine, low pole and non-polar component.
Use the liquid-phase conditions of polarity HILIC chromatographic columns for：UPLC BEH Amide HILIC column(2.1mm×
100mm, 1.7 μm), mobile phase is that A phase compositions are 95% acetonitrile and 5% aqueous solution containing 0.1% formic acid, B phase compositions be containing
There is the aqueous solution of 0.1% formic acid；Column temperature is 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：Just
Ion mode detection, mass range are arranged to 50-1200m/z full scan patterns.The optimal capillary of electro-spray ionization device
Tube voltage is 3000V, and Cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are
30L/h.Desolvation temperature is 400 DEG C, and ion source temperature is 100 DEG C.Concentration is used as 0.2ng/mL's
For leucineenkephalin (leucine enkephalin) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M
+H]+)。
Using low pole C18The liquid-phase condition of chromatographic column is：UPLC CSH C18column(2.1mm×100mm,1.7μ
M), mobile phase A phase composition is the aqueous solution containing 0.1% formic acid, and B phase compositions are acetonitrile；Column temperature is 40 DEG C, the μ L of sample size 2.0,
Flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：The two kinds of ionization mode detections of cation and anion, mass range are set
For 50-1200m/z full scan patterns.The optimal capillary voltage of electro-spray ionization device is 3000V (cation) or 2200V is (negative
Ion), Cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are 30L/h.Go molten
Agent temperature is 400 DEG C, and ion source temperature is 100 DEG C.Use leucineenkephalin (bright ammonia of the concentration for 0.2ng/mL
Sour enkephalins) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M+H]+) or 554.2615Da ([M-
H]-)。
Use the liquid-phase conditions of nonpolar PFPP chromatographic columns for：UPLC HSS PFPP column(2.1mm×100mm,
1.7 μm), mobile phase is that A phase compositions are the aqueous solution containing 0.1% formic acid, and B phase compositions are methanol；Column temperature is 40 DEG C, sample size
2.0 μ L, flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：The two kinds of ionization mode detections of cation and anion, quality model
Enclose and be arranged to 50-1200m/z full scan patterns.The optimal capillary voltage of electro-spray ionization device be 3000V (cation) or
2200V (anion), Cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are
30L/h.Desolvation temperature is 400 DEG C, and ion source temperature is 100 DEG C.Concentration is used as 0.2ng/mL's
For leucineenkephalin (leucine enkephalin) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M
+H]+) or 554.2615Da ([M-H]-)。
Each urine specimen detects polarity, low pole and non-polar component therein respectively using 3 kinds of chromatographic columns, wherein
Polar compound only detects positive ion mode, low pole and non-polar component detection cation and negative ion mode, i.e., each urine
Pattern detection 5 times, to detect metabolic compounds whole in urine as far as possible.Every 10 samples insert a sky in detection
It is white to prevent cross pollution, and insert a Quality Control sample and carry out quality control.Sample room temperature is kept for 4 DEG C in analyte detection process.
4th, data processing
The detection data of detection sample acquisition are analyzed using liquid chromatography/mass spectrometry combined instrument, metabolism group number is respectively adopted
Statistical analysis processing, including the extraction of alignment of data, peak are carried out according to processing professional software Progenesis QI.By the urine after work
The testing result of the testing result of liquid sample and the urine specimen before working work carries out pairing packet, using an orthogonal most young waiter in a wineshop or an inn partially
The detection data of multiplication (OPLS-DA) processing step 3, according to P≤0.05, CV≤30%, VIP>1.0th, maximum change multiple >=
1.2 as screening threshold value, and with reference to chromatogram retention characteristic and mass spectral characteristic, preliminary screening goes out characteristic biological mark.
In using polarity HILIC posts positive ion mode detection urine in the data of 7903 compounds of polar compound, point
Analysis filters out 4 kinds of compound being characterized property biomarkers；Utilizing low pole C18Weak pole in post positive ion mode detection urine
In the data of property 10463 compounds of component, Analysis and Screening goes out 5 kinds of compound being characterized property biomarkers；Utilize low pole
C18In post negative ion mode detection urine in the data of 6014 compounds of weakly polar component, Analysis and Screening goes out a kind of compound and is
Characteristic biological mark；9739 compounds of non-polar component in using nonpolar PFPP posts positive ion mode detection urine
Data in, Analysis and Screening goes out 4 kinds of compound being characterized property biomarkers；Examined using nonpolar PFPP posts negative ion mode
In survey urine in the data of 5996 compounds of non-polar component, fail Analysis and Screening and go out qualified characteristic biological mark
Thing.Concrete outcome is as shown in table 2.
5th, markers characteristic compares, and judges teams and groups' degree of fatigue
The characteristic biological mark filtered out in inspection table 2, contain air traffic control person teams and groups degree of fatigue
Related biomarker：Urocanic acid (Urocanic acid), acetylcytosine (N4-Acetylcytidine), 5- hydroxyl color ammonia
Sour (5-Hydroxy-L-tryptophan), dimethylguanosine (N2, N2-Dimehtylguanosine), antifebrin (N-
) and Alpha-CEHC Acetylarylamine.Further check each biomarker change multiple, urocanic acid (Urocanic
Acid) content declines 1.44 times, the decline of acetylcytosine (N4-Acetylcytidine) content 1.36 times, 5-hydroxyryptophan (5-
Hydroxy-L-tryptophan) under 1.62 times of content decline, dimethylguanosine (N2, N2-Dimehtylguanosine) content
1.50 times of drop, antifebrin (N-Acetylarylamine) content decline 1.78 times and Alpha-CEHC contents rise 1.33 times；
Judge the controller teams and groups day shift to be laid off when coming off duty accordingly and be in moderate fatigue state.
The characteristic fatigue degree associated biomarkers filtered out in the air traffic control person's urine specimen of table 2
Comparative example
1st, volunteer recruiting and sample collection
Volunteer recruiting：The people of administration and logistics personnel volunteer 23 is recruited in certain International airport at home, enters a group condition：Body is good for
Health, male, no medication, age 20-35 year；Sample collection was carried out autumn in October, 2016.
The collection of urine specimen：Once urinated in the day shift work pre-job collection of administration and logistics personnel volunteer working in 8 hours
Liquid sample After Hours gathers a urine specimen as tired sample as non-tired sample in administrative rear service personnel volunteer's day shift
This.Collect urine using Sterilized urine cup and be sub-packed in sterile tube and preserve.Urine specimen is sub-packed in sterile centrifugation tube and is stored in -80
℃。
2nd, the pretreatment of urine specimen
Urine specimen pre-treatment step is：- 80 DEG C of urine specimens will be stored in before analysis detection at 0-4 DEG C to melt, 4
DEG C, 12000rpm is centrifuged 5 minutes, then takes the μ L of supernatant 100 to add the dilution of 100 μ L water.
3rd, analysis detection is carried out to urine specimen using sewage sludge method
Polarity chromatographic column HILIC, low pole chromatographic column C is respectively adopted in the detection of urine specimen18With nonpolar chromatographic column
PFPP carries out separation detection respectively to the polarity in urine, low pole and non-polar component.
Use the liquid-phase conditions of polarity HILIC chromatographic columns for：UPLC BEH Amide HILIC column(2.1mm×
100mm, 1.7 μm), mobile phase is that A phase compositions are 95% acetonitrile and 5% aqueous solution containing 0.1% formic acid, B phase compositions be containing
There is the aqueous solution of 0.1% formic acid；Column temperature is 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：Just
Ion mode detection, mass range are arranged to 50-1200m/z full scan patterns.The optimal capillary of electro-spray ionization device
Tube voltage is 3000V, and Cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are
30L/h.Desolvation temperature is 400 DEG C, and ion source temperature is 100 DEG C.Concentration is used as 0.2ng/mL's
For leucineenkephalin (leucine enkephalin) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M
+H]+)。
Using low pole C18The liquid-phase condition of chromatographic column is：UPLC CSH C18column(2.1mm×100mm,1.7μ
M), mobile phase A phase composition is the aqueous solution containing 0.1% formic acid, and B phase compositions are acetonitrile；Column temperature is 40 DEG C, the μ L of sample size 2.0,
Flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：The two kinds of ionization mode detections of cation and anion, mass range are set
For 50-1200m/z full scan patterns.The optimal capillary voltage of electro-spray ionization device is 3000V (cation) or 2200V is (negative
Ion), Cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are 30L/h.Go molten
Agent temperature is 400 DEG C, and ion source temperature is 100 DEG C.Use leucineenkephalin (bright ammonia of the concentration for 0.2ng/mL
Sour enkephalins) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M+H]+) or 554.2615Da ([M-
H]-)。
Use the liquid-phase conditions of nonpolar PFPP chromatographic columns for：UPLC HSS PFPP column(2.1mm×100mm,
1.7 μm), mobile phase is that A phase compositions are the aqueous solution containing 0.1% formic acid, and B phase compositions are methanol；Column temperature is 40 DEG C, sample size
2.0 μ L, flow rate of mobile phase 0.3mL/min.Mass Spectrometry Conditions are：The two kinds of ionization mode detections of cation and anion, quality model
Enclose and be arranged to 50-1200m/z full scan patterns.The optimal capillary voltage of electro-spray ionization device be 3000V (cation) or
2200V (anion), Cone voltages are 30V.It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are
30L/h.Desolvation temperature is 400 DEG C, and ion source temperature is 100 DEG C.Concentration is used as 0.2ng/mL's
For leucineenkephalin (leucine enkephalin) as mass number calibration internal standard, calibration mass of ion is 556.2771Da ([M
+H]+) or 554.2615Da ([M-H]-)。
Each urine specimen detects polarity, low pole and non-polar component therein respectively using 3 kinds of chromatographic columns, wherein
Polar compound only detects positive ion mode, low pole and non-polar component detection cation and negative ion mode, i.e., each urine
Pattern detection 5 times, to detect metabolic compounds whole in urine as far as possible.Every 10 samples insert a sky in detection
It is white to prevent cross pollution, and insert a Quality Control sample and carry out quality control.Sample room temperature is kept for 4 DEG C in analyte detection process.
4th, data processing
The detection data of detection sample acquisition are analyzed using liquid chromatography/mass spectrometry combined instrument, metabolism group number is respectively adopted
Statistical analysis processing, including the extraction of alignment of data, peak are carried out according to processing professional software Progenesis QI.By the urine after work
The testing result of the testing result of liquid sample and the urine specimen before working work carries out pairing packet, using an orthogonal most young waiter in a wineshop or an inn partially
The detection data of multiplication (OPLS-DA) processing step 3, according to P≤0.05, CV≤30%, VIP>1.0th, maximum change multiple >=
1.2 as screening threshold value, and with reference to chromatogram retention characteristic and mass spectral characteristic, preliminary screening goes out characteristic biological mark.
In using polarity HILIC posts positive ion mode detection urine in the data of 8315 compounds of polar compound, point
Analysis filters out 10 kinds of compound being characterized property biomarkers；Utilizing low pole C18Weak pole in post positive ion mode detection urine
In the data of property 10645 compounds of component, Analysis and Screening goes out 17 kinds of compound being characterized property biomarkers；Utilize low pole
C18In post negative ion mode detection urine in the data of 6966 compounds of weakly polar component, Analysis and Screening goes out 7 kinds of compounds and is
Characteristic biological mark；10318 chemical combination of non-polar component in using nonpolar PFPP posts positive ion mode detection urine
In the data of thing, Analysis and Screening goes out 10 kinds of compound being characterized property biomarkers；Utilizing nonpolar PFPP posts anion mould
In formula detection urine in the data of 6571 compounds of non-polar component, fail Analysis and Screening and go out qualified characteristic biological
Mark.Concrete outcome is as shown in table 3.
5th, markers characteristic compares, and judges administration and logistics personnel's degree of fatigue
The characteristic biological mark filtered out in inspection table 3, only containing air traffic control person teams and groups fatigue journey
Spend related biomarker：Urocanic acid (Urocanic acid), acetylcytosine (N4-Acetylcytidine) and 5- hydroxyls
Tryptophan (5-Hydroxy-L-tryptophan).Further check each biomarker change multiple, urocanic acid (Urocanic
Acid) content declines 1.59 times, the decline of acetylcytosine (N4-Acetylcytidine) content 1.37 times, 5-hydroxyryptophan (5-
Hydroxy-L-tryptophan) content declines 1.43 times；Judge that administration and logistics personnel do not occur moderate fatigue accordingly；And accordingly
Judge administration and logistics personnel day shift work 8 hours it is next when characteristic biological mark species and changes of contents and civil aviaton it is empty
The species of characteristic biological mark before leaving offices and contain after middle traffic controller (day shift works 8 hours and made) teams and groups' day shift is laid off
Very big difference be present in amount change.
The characteristic fatigue degree associated biomarkers filtered out in the control group administration and logistics personnel's urine specimen of table 3
It should be noted that unless otherwise indicated, technical term or scientific terminology used in this application should be this hair
The ordinary meaning that bright one of ordinary skill in the art are understood.Unless specifically stated otherwise, otherwise illustrate in these embodiments
Part and relative step, numerical expression and the numerical value of step are not limit the scope of the invention.It is illustrated and described herein
In all examples, unless otherwise prescribed, any occurrence should be construed as merely exemplary, not as limitation, because
This, other examples of exemplary embodiment can have different values.
Finally it should be noted that：Various embodiments above is merely illustrative of the technical solution of the present invention, rather than its limitations；To the greatest extent
The present invention is described in detail with reference to foregoing embodiments for pipe, it will be understood by those within the art that：Its according to
The technical scheme described in foregoing embodiments can so be modified, either to which part or all technical characteristic
Carry out equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention skill
The scope of art scheme, it all should cover among protection scope of the present invention.
Claims (10)
1. a kind of air traffic control person teams and groups are slight and the detection method of moderate degree of fatigue, its characteristic are, wrap
Include following steps：
S1：The urine specimen of air traffic control person teams and groups is gathered, then the urine specimen is pre-processed；Its
In, the urine specimen includes the urine specimen before working work and the urine specimen after work；
S2：Separation detection is carried out using sewage sludge method respectively to the pretreated sample；Wherein,
The liquid chromatogram can use polarity HILIC chromatographic columns, low pole C18One kind in chromatographic column and nonpolar PFPP chromatographic columns
Or a variety of carry out separation detections；
S3：The inspection of urine specimen before the testing result of urine specimen after the teams and groups are worked and teams and groups working work
Survey result and carry out pairing comparison, filter out characteristic biological mark；
S4：By the characteristic biological mark of the screening biological marker related to air traffic control person teams and groups degree of fatigues
Thing is compared, and judges the degree of fatigue of air traffic control person.
2. air traffic control person teams and groups according to claim 1 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
In step S4, the biomarker bag slight to air traffic control person teams and groups and related moderate degree of fatigue
Include but be not limited to：Urocanic acid (Urocanic acid, C6H6N2O2, HMDB 00301), acetylcytosine (N4-
Acetylcytidine, C11H15N3O6, HMDB 05923), 5-hydroxyryptophan (5-Hydroxy-L-tryptophan,
C11H12N2O3, HMDB 00472), dimethylguanosine (N2, N2-Dimehtylguanosine, C12H17N5O5, HMDB 04284),
Antifebrin (N-Acetylarylamine, C8H9NO, HMDB 01250) and Alpha-CEHC (C16H22O4, HMDB 01518).
3. air traffic control person teams and groups according to claim 2 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
In step S4, in the characteristic biological mark screened in step s3, urocanic acid, acetylcytosine, 5- hydroxyl colors are searched
Propylhomoserin, dimethylguanosine, antifebrin and Alpha-CEHC, the species occurred according to features described above mark and its change model
Enclose, can determine whether that slight or moderate fatigue occur in air traffic control person teams and groups.
4. air traffic control person teams and groups according to claim 3 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
In step S3, the testing result of the urine specimen after the 2-3 hours that preferably worked by day after teams and groups' work and the class
The testing result paired comparisons of urine specimen before group working work, in the markers characteristic filtered out, if detection urine periodical
Acid, and the up-regulation of its content meets or exceeds 1.4 times, you can judge that slight fatigue occur in air traffic control person teams and groups；
Preferably, the testing result for the urine specimen after 3-5 hours that preferably worked by day after teams and groups work and the teams and groups
The testing result paired comparisons for the urine specimen before work of going to work, in the markers characteristic filtered out, if detection urocanic acid,
Acetylcytosine, 5-hydroxyryptophan, and the decline of their contents respectively reaches and more than more than 1.4,1.3,1.4 times, you can sentence
There is moderate fatigue in disconnected air traffic control person teams and groups；
Preferably, the testing result for the urine specimen after 5-8 hours that preferably worked by day after teams and groups work and the teams and groups
The testing result paired comparisons for the urine specimen before work of going to work, in the markers characteristic filtered out, if detection urocanic acid,
Acetylcytosine, 5-hydroxyryptophan, dimethylguanosine, antifebrin and Alpha-CEHC, and first five kind content declines difference
Reach and more than more than 1.4,1.3,1.4,1.4,1.4 times and Alpha-CEHC contents rise reach and more than more than 1.3 times, i.e.,
It can determine whether that moderate fatigue occur in air traffic control person teams and groups.
5. air traffic control person teams and groups according to claim 1 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
In step S1, the urine specimen pretreatment includes：The urine specimen for being stored in -80 DEG C is melted at 0-4 DEG C, then
At 4 DEG C, 12000rpm centrifuges the urine specimen 5min, and supernatant is collected after centrifugation；Supernatant described in 100 μ L is taken, so
Diluted afterwards with 100 μ L water.
6. air traffic control person teams and groups according to claim 1 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
In step S2, sample room temperature preferably keeps 0-4 DEG C during the separation detection；The liquid chromatogram uses polarity
During HILIC chromatographic column separation detection polar compounds, mass spectrography is using detection cation ionization mode；The liquid chromatogram uses
Low pole C18When chromatographic column separation detection low pole and non-polar component, mass spectrography is using the two kinds of ionizations of cation and anion
Pattern；When the liquid chromatogram uses nonpolar PFPP chromatographic columns separation detection low pole and non-polar component, mass spectrography uses
Two kinds of ionization modes of cation and anion.
7. air traffic control person teams and groups according to claim 6 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
In step S2, when the liquid chromatogram uses polarity HILIC chromatographic column separation detection polar compounds, mass spectrography is using detection
Cation ionization mode, testing conditions are preferably：Mass range is arranged to 50-1200m/z full scan patterns；Electron spray ion
It is 3000V to change the optimal capillary voltage of device, and Cone voltages are 30V；It is nitrogen to dry gas, and desolvation flow velocity is 800L/h,
Cone gas velocities are 30L/h；Desolvation temperature is 400 DEG C, and ion source temperature is 100 DEG C；Concentration is used as 0.2ng/mL's
Leucine enkephalin calibrates internal standard as mass number, and calibration mass of ion is 556.2771Da ([M+H]+)；Liquid-phase condition is：
UPLC BEH Amide HILIC column(2.1mm×100mm,1.7μm)；Mobile phase includes A phases and B phases, the A phases group
Be 5% aqueous solution containing 0.1% formic acid as the acetonitrile of volume fraction 95% and volume fraction, the B phase compositions be containing
The aqueous solution of 0.1% formic acid；Column temperature is 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.
8. air traffic control person teams and groups according to claim 6 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
In step S2, the liquid chromatogram uses low pole C18When chromatographic column separation detection low pole and non-polar component, mass spectrum
Method uses two kinds of ionization modes of cation and anion, and testing conditions are preferably：It is complete that mass range is arranged to 50-1200m/z
Scan pattern；The optimal capillary voltage of electro-spray ionization device is 3000V (cation) or 2200V (anion), cone voltages
For 30V；It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are 30L/h；Desolvation temperature is 400
DEG C, ion source temperature is 100 DEG C；Concentration is used to calibrate internal standard, calibration as mass number for 0.2ng/mL leucine enkephalin
Mass of ion is 556.2771Da ([M+H]+) or 554.2615Da ([M-H]-)；Liquid-phase condition is：UPLC CSH C18column
(2.1mm×100mm,1.7μm)；Mobile phase A phase composition is the aqueous solution containing 0.1% formic acid, and B phase compositions are acetonitrile；Column temperature
For 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.
9. air traffic control person teams and groups according to claim 6 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
In step S2, when the liquid chromatogram uses nonpolar PFPP chromatographic columns separation detection low pole and non-polar component, matter
Spectrometry uses two kinds of ionization modes of cation and anion, and testing conditions are preferably：Mass range is arranged to 50-1200m/z
Full scan pattern；The optimal capillary voltage of electro-spray ionization device is 3000V (cation) or 2200V (anion), cone electricity
Press as 30V；It is nitrogen to dry gas, and desolvation flow velocity is 800L/h, and cone gas velocities are 30L/h；Desolvation temperature is 400
DEG C, ion source temperature is 100 DEG C；Concentration is used to calibrate internal standard, calibration as mass number for 0.2ng/mL leucine enkephalin
Mass of ion is 556.2771Da ([M+H]+) or 554.2615Da ([M-H]-)；Liquid-phase condition is：UPLC HSS PFPP
column(2.1mm×100mm,1.7μm)；Mobile phase is that A phase compositions are the aqueous solution containing 0.1% formic acid, and B phase compositions are first
Alcohol；Column temperature is 40 DEG C, sample size 2.0 μ L, flow rate of mobile phase 0.3mL/min.
10. air traffic control person teams and groups according to claim 6 are slight and the detection method of moderate degree of fatigue,
Its characteristic is：
Step S3 is specifically included：The detection data that will be obtained using the detection of sewage sludge method, at data
Manage software, it is preferred to use metabolism group data processing related software carries out statistical analysis processing, further preferably uses metabolism group
Learn data processing arts software Pro genesis QI and carry out statistical analysis processing；
The analysis and processing method includes alignment of data, peak extraction；Also include to air traffic control person teams and groups work
The inspection of urine specimen before the testing result of urine specimen after work and air traffic control person teams and groups working work
Survey result and carry out pairing packet, then using PLS OPLS-DA Treatment Analysis, with P≤0.05, CV≤30%, VIP
>1.0th, maximum change multiple >=1.2 filter out characteristic biological mark as screening threshold value.
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