CN104614291B - Assess mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity - Google Patents
Assess mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity Download PDFInfo
- Publication number
- CN104614291B CN104614291B CN201510016660.3A CN201510016660A CN104614291B CN 104614291 B CN104614291 B CN 104614291B CN 201510016660 A CN201510016660 A CN 201510016660A CN 104614291 B CN104614291 B CN 104614291B
- Authority
- CN
- China
- Prior art keywords
- reference levels
- mark
- particulate matter
- fine particulate
- linolenyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity are assessed the present invention relates to a kind of; the mark includes the one or more in dihydrosphingosine, phytosphingosine, DSPC, the phosphocholine of linolenyl glyceryl 3, the phosphocholine of linolenyl glyceryl 4, palmitamide, valine, cytimidine; the present invention can more comprehensively, synthetically embody the changing condition that airborne fine particulate matter exposure causes organism metabolite, and the assessment of toxic action is exposed suitable for airborne fine particulate matter.
Description
Technical field
The present invention relates to mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity is assessed, belong to air
The detection field of fine particle toxicity.
Background technology
In China, airborne fine particulate matter (PM2.5) pollution is the health risk factors of ranking the 4th, and first 3 are high blood respectively
Pressure, smoking and poor eating habits.Fine particle PM2.5 refers to suspension of the aerodynamic diameter less than 2.5 μm in air
Grain thing, compared with thicker Atmospheric particulates, PM2.5 particle diameters are small, and specific surface area is big, are adsorbed with more harmful substances, and big
Residence time length, fed distance are remote in gas, are more easy to enter alveolar and be deposited in deep lung to be not easy to be discharged by human body, thus to big
The influence of gas environmental quality and health is bigger.According to US Adm Nati in the whole world that in September, 2010 is announced
PM2.5 pollution concentration map denotations, China especially North China, east and central China area are the severely afflicated areas polluted.
PM2.5 influences the correlative study of breathing and cardiovascular system by wide coverage, the interference of its Endocrine system
It is current new study hotspot.Nearest epidemiological study shows that long-term PM2.5 exposures are that fat and diabetes are occurred frequently
The significant contribution factor.The harmful substances such as PM2.5 particle surfaces attachment heavy metal, polycyclic aromatic hydrocarbon, volatile organic matter, wherein a lot
Component all has E＆M disturbing effect.PM2.5 exposures cause human endocrine and metabolic disorder, this disorderly possible
Induce the diseases such as fat and diabetes.Bacterium, virus, heavy metal, acidic oxide and the organic pollution of PM2.5 attachments are to people
The degree of injury of body has very big difference, according to current passing method, can not accurate evaluation PM2.5 to the detrimental extent of human body.
A large amount of evidences demonstrate Atmospheric particulates really can Endocrine system produce interference effect, and this interference is imitated
Assessment should be carried out to have great importance.The disorder of internal system can cause metabolic disorder, so as to induce a variety of endocrine generations
Thanking property disease.There is complicated crossedpath in E＆M control process, this needs to develop more high flux, accurate and have
The means of effect evaluate the disturbing effect of pollutant.Moreover, PM2.5 fine graineds are except containing heavy metal, polycyclic aromatic hydrocarbon, performance property
Outside the compositions such as organic matter, also containing compositions such as organic carbon, elemental carbons, therefore PM2.5 disturbing effect is different from Single Pollution thing,
Influence to organism metabolic system is extremely complex, and existing biology theory and method expose larger in terms of analysis throughput
Limitation.
The change of the genome and apparent gene of organism, will necessarily influence protein caused by environmental contaminants exposure
Expression, finally further amplifies in the aspect of terminal metabolin.Metabolism group is emerging omics technology, by existing to organism
All metabolins carry out qualitative and quantitative determination simultaneously under specific poisonous substance stimulates, and pattern-recognition is carried out to overall metabolic profile, from
And poisonous effect evaluation is carried out, and disclose the target organ and tissue, toxicity process of toxic action.High-throughout metabolism group
Technology can find the changing rule of metabolin after PM2.5 exposures from integral level, so as to be ground to its disturbing effect
Study carefully, and characteristic metabolic thing can be filtered out and be used for health risk assessment as Exposure effect mark.With gene and protein
Compare, metabolin is simple and easy to get, is closely connected with the terminal of traditional toxicological effect, has as Exposure effect mark
Its only thick advantage.
The content of the invention
The technical problems to be solved by the invention, which are to provide, assesses what airborne fine particulate matter exposure was acted on organismal toxicity
Mark and method, the present invention can more comprehensively, synthetically embody airborne fine particulate matter exposure and cause organism metabolite
Changing condition, the assessment of toxic action degree is exposed suitable for airborne fine particulate matter.
The technical scheme that the present invention solves above-mentioned technical problem is as follows：Airborne fine particulate matter exposure is assessed to organismal toxicity
The mark of effect, it is sweet that the mark includes dihydrosphingosine, phytosphingosine, DSPC, linolenyl
Oil base -3- phosphocholines, linolenyl glyceryl -4- phosphocholines, palmitamide, valine, one kind in cytimidine or several
Kind.
The technical scheme that the present invention solves above-mentioned technical problem is as follows：Airborne fine particulate matter exposure is assessed to organismal toxicity
The method of effect, including：
200 μ L experimenter's serums are taken, the pure methanol of 3 times of volumes is added, acutely concussion 5 minutes, stands 5 minutes, centrifugation 15
Minute, clear liquid is transferred in centrifuge tube, it is concentrated into absolutely dry, the dissolving of 200 μ L pure methanol is added, passes through liquid chromatogram-matter
GC-MS is composed, detects total finger-print of experimenter's serum, calculates the normalizing of at least one of experimenter's serum mark
Change area, by result with the reference levels of corresponding mark compared with, difference instruction airborne fine particulate matter expose whether to by
Examination person produces toxic action,
Wherein, the mark includes dihydrosphingosine, phytosphingosine, DSPC (also known as haemolysis ovum
Phosphatide (18:0)), linolenyl glyceryl -3- phosphocholines (also known as lysolecithin (18:3)), linolenyl glyceryl -4-
Phosphocholine (also known as lysolecithin (18:4)), the one or more in palmitamide, valine, cytimidine,
The reference levels refer to the mark being not exposed in PM2.5 experimenter's serum in the total fingerprint chromatogram of serum
Normalized area, dihydrosphingosine reference levels are 0.5-1%, and phytosphingosine reference levels are 1-1.4%, stearoyl phosphatidyl
Phatidylcholine reference levels are 1-2%, and linolenyl glyceryl -3- phosphocholines reference levels are 0.2-0.3%, and linolenyl is sweet
Oil base -4- phosphocholines reference levels are 0.5-1%, and palmitamide reference levels are 0.8-1.1%, and valine reference levels are
0.5-0.9%, cytimidine reference levels are 0.3-0.5%.
On the basis of above-mentioned technical proposal, the present invention can also do following improvement.
Further, the subject is mammal.
Further, the centrifugal force is 12000g.
A kind of method of the mark acted on the invention also discloses screening assessment airborne fine particulate matter organismal toxicity,
Including：
(a) all inspections in airborne fine particulate matter exposed population group's blood serum sample are determined by LC-MS-MS
The relative concentration levels of the metabolin measured, as detection group；
All detections in crowd's serum of airborne fine particulate matter are not exposed to by LC-MS-MS measure
The relative concentration levels of the metabolin arrived, as a control group；
(b) the suitable mark for characterizing toxicity of chemometrics method screening is combined：Idiographic flow is using orthogonal inclined
Minimum variance techniques of discriminant analysis is directed to training set Sample Establishing model, using the VIP scores of variable as the potential mark of screening
Superset a key factor, VIP scores are selected, then, are examined using nonparametric in this step more than 1.5 variable
Test as final program, exclude the variable that p value is more than 0.05.
(c) by a pair of the relative concentration levels 1 of all metabolins detected in detection group and control group blood serum sample
It should be compared, show that the metabolin of difference is accredited as indicating the mark of airborne fine particulate matter exposure toxic action.
The invention discloses the method that assessment airborne fine particulate matter exposure is acted on organismal toxicity, it is related to blood serum sample
Analytical test.In certain aspects, the mark for assessing airborne fine particulate matter exposure toxic action is contained, includes use
In the method identified and detect mark.
The present invention is used as a kind of evaluation measures, realizes to the simple of airborne fine particulate matter exposure toxic action and accurately sentences
It is disconnected.Specifically have the beneficial effect that：
(1) sample needed for detection is serum, and it is convenient to obtain, and avoids harmful complicated mistake such as body injury, radial imaging
Journey；
(2) as a kind of new appraisal procedure, airborne fine particulate matter exposure actual poison to caused by body is accurately reflected
Property effect.
(3) present invention, which realizes, more comprehensively, synthetically embodies the change that airborne fine particulate matter exposure causes organism metabolite
Change situation, expose toxic action scale evaluation suitable for airborne fine particulate matter.
Embodiment
The principles and features of the present invention are described below, and the given examples are served only to explain the present invention, is not intended to limit
Determine the scope of the present invention.
Embodiment
Sample collection
The application have collected the serum sample of two class crowds, and first kind crowd (n=230) lives in pm2.5 and seriously polluted
Area (have 240 days pm2.5 concentration every year>150μg/m3), it is relatively low that the second class crowd (n=210) lives in pm2.5 pollutions
(there is 10 days pm2.5 concentration every year>25μg/m3) area.80% random sample (first kind crowd n=in each crowd
184；Second class crowd n=168) establish statistical models, remaining 20% sample (first kind crowd n=46；Second class crowd
N=42) as checking sample.
UPLC-oa-TOF-MS conditions
Blood serum metabolic collection of illustrative plates passes through Ultimate-3000 liquid chromatographs and MicrOTOF-Q II- level Four bar flight time
Mass spectrometry system obtains.Reverse phase separation uses the Phenomenex C18 colors that filler aperture is 2.1 × 100mm1.7 μm
Post is composed, column temperature maintains 35 DEG C, and sampling volume is 5 μ L.Mobile phase is (A) 0.1% aqueous formic acid and (B) 0.1% first
The mixed liquor of sour acetonitrile solution, gradient are as follows：Within 1-16 minutes, Mobile phase B volume fraction rises to 100% by 5%,
In 16-20 minutes, Mobile phase B is 100% and kept for 4 minutes, then 5% was dropped in 0.1 minute, final Mobile phase B volume
Fraction maintains 3 minutes 5%.Flow rate of mobile phase is 400 μ L/min.Mass spectrograph is run under cation, ESI scan patterns, is swept
Retouch scope and arrive 1000m/z for 50.Data are collected under centroid patterns.Capillary voltage and end plate drift potential are set respectively
For 4500V and -500V.Spray pressure is arranged to 0.6bar, dry gas flow 6L/min, and temperature is 200 DEG C.Utilize second order mses
The further potential mark of experiment.Collision gas is argon gas, and for each mark, impact energy is 30eV.
Data analysis
All chromatograms collected are carried out with peak alignment, background noise removal and retention time-molecular mass ion pair
Extraction.The sample of 20% variable missing is given up, and is then introduced into SIMCA-P v12 softwares by normalization and Pareto medellings
Afterwards, multivariate data analysis is carried out.Training set Sample Establishing model is directed to using orthogonal minimum variance techniques of discriminant analysis partially, to air
Fine particle exposure group and control group are distinguished and screen mark, and are further confirmed with non-parametric test method, and P values are less than
0.05 is thought with the significance of difference statistically.
Mark is identified
Using the possibility molecular formula of the software fitting token things of DataAnalysis 4.0, and structural information is submitted into Human
Urine Metabolome Database (HMDB) database retrieval, candidate markers are carried out with second mass analysis, it is final logical
Cross compared with the standard items of commercialization, with final acknowledgement indicator thing.
Statistical analysis
Statistical analysis is carried out using the softwares of SPSS Statistics 18 (SPSS Inc.).Using Kruskal-Wallis
Method of inspection deletes the potential mark for selecting metabolin to have significant change in three dosage groups.Using Wilcoxon signed rank test methods
Compare the marker levels between airborne fine particulate matter exposure group and control group, p value is less than the 0.05 instruction significance of difference.
The screening of mark
Metabolic profile difference between case group and control group is maximized using the OPLS-DA for having supervision.Selection is to two groups
Between the maximum variable of classification contribution as potential mark.First, using the VIP scores of variable as the potential mark of screening
One key factor of the superset of will thing.Variable of the VIP scores more than 1.5 is selected in this step.Then, using non-ginseng
Number, which is examined, is used as final program, excludes the variable that p value is more than 0.05.
Further using Receiver operating curve (receiver operating characteristic curve,
Abbreviation ROC curve) diagnosis performance of these marks clinically is assessed.TG-AUC (area
Under curve) it is important parameter in ROC models, the diagnosis performance of the index characterization mark, it is however generally that, AUC>
0.75 represents that the index has good diagnosis capability.As shown in table 1, the AUC of all marks is above 0.75, represents
They all have good evaluation performance to the health effect of Atmospheric particulates exposure.
The mark of table 1
aVIP comes from OPLS-DA models, threshold value 1.5.
bP values are calculated by non-parametric test.
cChange multiple and indicate that concentration of the mark in case group is of a relatively high more than 1, and the mark is indicated less than 1
Thing concentration is relatively low in control group.
dTG-AUC (area under curve).
The checking of model
We use checking sample (first kind crowd n=46；Second class crowd n=42) to the model and mark established
Thing is verified that correct differentiation can be obtained by finding more than 90% sample standard deviation.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent substitution and improvements made etc., it should be included in the scope of the protection.
Claims (3)
1. assess the method that airborne fine particulate matter exposure is acted on organismal toxicity, it is characterised in that including：
200 μ L experimenter's serums are taken, the pure methanol of 3 times of volumes is added, acutely concussion 5 minutes, stands 5 minutes, centrifuge 15 minutes,
Clear liquid is transferred in centrifuge tube, it is concentrated into absolutely dry, the dissolving of 200 μ L pure methanol is added, passes through liquid chromatograph mass spectrography
Technology, total finger-print of experimenter's serum is detected, calculates the normalized area of at least one of experimenter's serum mark,
By result compared with the reference levels of corresponding mark, whether difference instruction airborne fine particulate matter exposure produces to subject
Toxic action,
Wherein, the mark include dihydrosphingosine, phytosphingosine, DSPC, linolenyl glyceryl-
One or more in 3- phosphocholines, linolenyl glyceryl -4- phosphocholines, palmitamide, valine, cytimidine,
The reference levels refer to normalizing of the mark being not exposed in PM2.5 experimenter's serum in the total fingerprint chromatogram of serum
Change area, dihydrosphingosine reference levels are 0.5-1%, and phytosphingosine reference levels are 1-1.4%, stearyl phosphatidyl courage
Alkali reference levels are 1-2%, and linolenyl glyceryl -3- phosphocholines reference levels are 0.2-0.3%, linolenyl glycerine
Base -4- phosphocholines reference levels are 0.5-1%, and palmitamide reference levels are 0.8-1.1%, and valine reference levels are
0.5-0.9%, cytimidine reference levels are 0.3-0.5%.
2. according to the method for claim 1, it is characterised in that the subject is mammal.
3. according to the method for claim 1, it is characterised in that the centrifugal force is 12000g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510016660.3A CN104614291B (en) | 2015-01-13 | 2015-01-13 | Assess mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510016660.3A CN104614291B (en) | 2015-01-13 | 2015-01-13 | Assess mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104614291A CN104614291A (en) | 2015-05-13 |
| CN104614291B true CN104614291B (en) | 2018-01-16 |
Family
ID=53148837
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510016660.3A Expired - Fee Related CN104614291B (en) | 2015-01-13 | 2015-01-13 | Assess mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104614291B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108351342B (en) * | 2015-08-20 | 2021-04-02 | 深圳华大生命科学研究院 | Biomarkers for coronary heart disease |
| CN105628809B (en) * | 2015-12-25 | 2018-04-03 | 中国药科大学 | The metabolic markers of coronary atherosclerosis and coronary heart disease are distinguished for diagnosing |
| CN105486773A (en) * | 2015-12-25 | 2016-04-13 | 齐炼文 | Metabolic marker for diagnosing coronary heart disease |
| CN105445408B (en) * | 2016-01-25 | 2018-06-12 | 齐炼文 | The metabolic markers of coronary atherosclerosis and stable angina cordis are distinguished in diagnosis |
| CN107589200B (en) * | 2017-09-15 | 2020-06-16 | 山东省科学院生物研究所 | Method for evaluating early toxicity of trace exogenous chemicals by using diagnostic marker and application thereof |
| CN109839498A (en) * | 2017-11-27 | 2019-06-04 | 中国科学院大连化学物理研究所 | A kind of appraisal procedure that PM2.5 particle influences Radical Metabolism access in human pneumonocyte |
| CN115372521B (en) * | 2022-09-21 | 2024-07-26 | 重庆智合生物医药有限公司 | Method for separating and identifying phytosphingosine and/or N-acetylphytosphingosine |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5397894A (en) * | 1993-05-28 | 1995-03-14 | Varian Associates, Inc. | Method of high mass resolution scanning of an ion trap mass spectrometer |
| CN103814295A (en) * | 2011-09-14 | 2014-05-21 | 巴斯夫欧洲公司 | Means and methods for assessing kidney toxicity |
-
2015
- 2015-01-13 CN CN201510016660.3A patent/CN104614291B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5397894A (en) * | 1993-05-28 | 1995-03-14 | Varian Associates, Inc. | Method of high mass resolution scanning of an ion trap mass spectrometer |
| CN103814295A (en) * | 2011-09-14 | 2014-05-21 | 巴斯夫欧洲公司 | Means and methods for assessing kidney toxicity |
Non-Patent Citations (7)
| Title |
|---|
| AD患者血清中L-谷氨酰胺、溶血磷脂酰胆碱和棕榈酰胺HPLC-MS测定;解微 等;《中国公共卫生》;20140530;第30卷(第5期);第691-692页 * |
| Identification of Human Plasma Metabolites Exhibiting Time-of-Day Variation Using an Untargeted Liquid Chromatography-Mass Spectrometry Metabolomic Approach;Joo Ern Ang，et al.;《Chronobiology International》;20120723;第29卷(第7期);第872页表1 * |
| 基于LC-MS-代谢组学技术在慢性肾脏病的应用;冯亚龙 等;《药物分析杂志》;20131231;第33卷(第10期);第1812页第2.1节第2段 * |
| 基于代谢组学技术应激抑郁动物模型的评价;郑兴宇 等;《药物评价研究》;20100630;第33卷(第3期);摘要 * |
| 基于超高效液相色谱质谱的药物性肝损害患者血清代谢组学研究;李小芬;《中国微生态学杂志》;20130630;第25卷(第6期);摘要以及第2.3节 * |
| 自发性腹膜炎的代谢学研究;牛玲玲;《中国优秀硕士学位论文全文数据库 医药卫生科技辑（月刊）》;20120915(第09期);摘要第III页第2-5段 * |
| 酵母菌添加对饲喂不同精粗比饲粮肉牛瘤胃发酵、养分降解和血浆代谢组的影响;丁耿芝;《中国博士学位论文全文数据库 农业科技辑（月刊）》;20140815(第06期);摘要第II页第2段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104614291A (en) | 2015-05-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104614291B (en) | Assess mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity | |
| CN109884302A (en) | Lung cancer early diagnosis marker and its application based on metabolism group and artificial intelligence technology | |
| Bajoub et al. | Comparing two metabolic profiling approaches (liquid chromatography and gas chromatography coupled to mass spectrometry) for extra-virgin olive oil phenolic compounds analysis: A botanical classification perspective | |
| Møller et al. | Oxidative damage to DNA and lipids as biomarkers of exposure to air pollution | |
| CN106093227A (en) | The LC-MS method of 113 kinds of lipids in a kind of high flux detection organism blood sample | |
| CN110057955A (en) | The screening technique of hepatitis B specific serum marker | |
| Li et al. | Urinary metabolomics revealed arsenic exposure related to metabolic alterations in general Chinese pregnant women | |
| CN110082444A (en) | The construction method of the mouse model for screening particulate matter exposure early effect marker based on lipid composition analysis | |
| CN101772701A (en) | Carry out lipoprotein analysis by differential charged-particle mobility | |
| CN110178035B (en) | Type 2 diabetes marker and application thereof | |
| Zhang et al. | High-throughput and high-sensitivity quantitative analysis of serum unsaturated fatty acids by chip-based nanoelectrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry: early stage diagnostic biomarkers of pancreatic cancer | |
| CN105738526A (en) | Method for screening specific serum metabolism markers for triple-negative breast cancer | |
| CN103630678A (en) | Biological marker of male infertility and application thereof | |
| CN106526028A (en) | Applications of metabolic markers in diagnosing and identifying benign or malignant lesions of thyroid gland | |
| Zhang et al. | Effects of the ambient fine particulate matter (PM2. 5) exposure on urinary metabolic profiles in rats using UPLC-Q-TOF-MS | |
| CN110702821B (en) | Typing detection kit for chronic obstructive pulmonary disease | |
| Gong et al. | Applications of in vivo SPME based on mass spectrometry for environmental pollutants analysis and non-target metabolomics: A review | |
| CN110687231A (en) | Typing detection kit for chronic obstructive pulmonary disease | |
| CN109946411B (en) | Biomarker for diagnosis of ossification of yellow ligament of thoracic vertebra and screening method thereof | |
| CN104297442B (en) | The application in terms of quickly detection early stage cardiac toxicity of the endogenous small-molecule substance | |
| CN103776914A (en) | Biomarker of male infertility in unknown aetiology and application method of biomarker | |
| CN111289678A (en) | Rhizoma anemarrhenae quality detection method based on UPLC-QQQ-MS/MS method | |
| CN109946467B (en) | Biomarker for ossification diagnosis of thoracic vertebra ligamentum flavum | |
| CN110327131A (en) | Tissue metabolism's network research method of Atmospheric particulates poisonous effect | |
| WO2011003922A1 (en) | Method for the diagnosis of asthma by detecting volatile organic compounds in exhaled air |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180116 Termination date: 20220113 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |