CN110487929A - A method of utilizing trimethylamine oxide content in reaction assay human serum in ml headspace bottle - Google Patents
A method of utilizing trimethylamine oxide content in reaction assay human serum in ml headspace bottle Download PDFInfo
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- CN110487929A CN110487929A CN201910757106.9A CN201910757106A CN110487929A CN 110487929 A CN110487929 A CN 110487929A CN 201910757106 A CN201910757106 A CN 201910757106A CN 110487929 A CN110487929 A CN 110487929A
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- China
- Prior art keywords
- headspace bottle
- human serum
- trimethylamine oxide
- trimethylamine
- oxide content
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/16—Injection
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/065—Preparation using different phases to separate parts of sample
Abstract
The present invention relates to it is a kind of using in ml headspace bottle in reaction assay human serum trimethylamine oxide content method, it is that human serum is injected into ml headspace bottle, it sequentially adds reducing agent and vaporization agent and trimethylamine oxide in serum is reduced to trimethylamine in a heated condition and is vaporized, using gas Chromatographic Determination trimethylamine, the content of trimethylamine oxide thus can be obtained indirectly.The present invention low, easy to operate, high sensitivity, high repeatability and other advantages with cost of determination.It can effectively solve the problems, such as directly utilize trimethylamine oxide content in gas Chromatographic Determination serum.
Description
Technical field
The invention belongs to technical field of medical detection, are related to a kind of utilize in ml headspace bottle and aoxidize in reaction assay human serum
The method of trimethylamine content.
Background technique
Choline can play as mankind's essential nutrients and adjust lipid metaboli, the prevention and treatment diseases such as fatty liver and cardiovascular disease
The effects of with neurodevelopment is promoted.But choline levels are increased with the occurrence risk of metabolic syndrome, cardiovascular disease etc. in just
Correlation, this relies on the lower trimethylamine oxide close relation generated of intestinal flora effect with choline.Internal high level of oxidation three
Methylamine can influence cholesterol metabolic, atherosclerotic plaque is promoted to form and aggravate disorders of lipid metabolism.Therefore, vivo oxidation three is measured
Methylamine content has to the relationship that metabolic condition and choline of the choline in enteron aisle rely on intestinal flora metabolism object and chronic disease is inquired into
Important meaning.
The chemical structure of trimethylamine oxide is similar to methyl donor such as choline, glycine betaine and S-adenosylmethionine, with hat
The close correlation of the diseases such as heart trouble, cardiovascular disease, atrial fibrillation and atheroma.Trimethylamine oxide in normal human blood
0.73~126 μm of ol/l of content, the method for measuring trimethylamine oxide is related to using large-scale precision and expensive instrument, such as mass spectrum
Method, High Performance Liquid Chromatography/Mass Spectrometry on-line method etc..These methods are complicated for operation, and higher cost cannot be widely popularized.Existing measurement
In the method for trimethylamine oxide content, the Sensitivity of Absorption Photometry is low, low efficiency, poor repeatability；Gas chromatography high sensitivity,
It is fast to analyze speed, is suitable for trace analysis, but trimethylamine oxide is not volatile；Ion chromatography analysis trimethylamine oxide speed
Fastly, but Common Cations interference is larger；Liquid chromatography needs first by its derivatization treatment；Capillary electrophoresis detection is limited to
Low, high sensitivity, but since capillary diameter is small, optical path is short, poor repeatability.
Summary of the invention
It is an object of the invention in view of the above shortcomings of the prior art, provide a kind of utilize to react generation in ml headspace bottle
The method of trimethylamine oxide content in trimethylamine indirect determination human serum, gas Chromatographic Determination blood can not directly be utilized by solving
In clear the problem of trimethylamine oxide content.
The purpose of the present invention is what is be achieved through the following technical solutions:
A method of using trimethylamine oxide content in reaction assay human serum in ml headspace bottle, using anti-in ml headspace bottle
Trimethylamine should be generated, gas Chromatographic Determination is carried out and obtains the content of trimethylamine oxide, comprising the following steps:
A, human serum is injected into ml headspace bottle, reducing agent and vaporization agent is added and heated, will aoxidized in human serum
Trimethylamine is reduced to trimethylamine and vaporizes；
B, the gas for being taken certain volume from ml headspace bottle using head-space sampler is surveyed through gas-chromatography separation, fid detector
Determine trimethylamine, calculates the content of trimethylamine oxide.
Further, step A, the amount that human serum is added are the 1/8-1/4 of ml headspace bottle volume.
Further, step A, the reducing agent are titanium trichloride, and vaporization agent is saturation potassium hydroxide solution, and ml headspace bottle is held
Product: reducing agent volume: vaporization agent volume is 4:0.05~0.12:0.1~0.5.
Further, step A, the ml headspace bottle reaction temperature are 60~95 DEG C, and the reaction time is greater than 2min.
Further, step B can use quantified by external standard method measurement trimethylamine oxide, also can use internal standard method (internal standard
Compound is isopropylamine) quantitative determination trimethylamine oxide.
Compared with prior art, the present invention utilizes the side of trimethylamine oxide content in reaction assay human serum in ml headspace bottle
Method, low, easy to operate, high sensitivity, high repeatability and other advantages with cost of determination；For assessment coronary heart disease, cardiovascular and cerebrovascular disease,
The risk of the diseases such as atrial fibrillation and atheroma provides the measuring method of trimethylamine oxide content, realizes the heart
Risk assessment, screening and the prevention of vascular diseases.
Detailed description of the invention
Fig. 1 is the chromatogram of gas chromatograph for determination after trimethylamine oxide restores in ml headspace bottle.
Fig. 2 is the concentration relationship of chromatography peak height and trimethylamine oxide.
Specific embodiment
Embodiment 1
It takes certain volume (0.5~1ml) human serum sample in 4ml ml headspace bottle, 0.1ml titanium trichloride solution is added
(20%), after sealing water-bath 5 minutes in 70 DEG C of water, it is then injected into 0.5ml saturation potassium hydroxide solution, is continued in 70 DEG C of water
Water-bath 5 minutes, headspace gas 1ml is taken to carry out chromatographic isolation measurement from ml headspace bottle using head-space sampler.Chromatographic condition is to carry
Gas: it is pressed before high pure nitrogen (99.999%), column: 0.05MPa, gasification temperature: 180 DEG C, column temperature: 120 DEG C, detector: FID, inspection
180 DEG C of testing temperature, chromatographic column: PEG-20M capillary column.
The trimethylamine oxide chromatographic peak obtained under above-mentioned chromatographic condition, retention time 2.365min, blood serum sample color
Spectrogram is as shown in Figure 1.
Using the above method, will be respectively 0 with the concentration of human serum sample's same volume, 10,50,100,150,200,
250 μm of ol/l trimethylamine oxide solution carry out same head space reaction respectively and chromatographic isolation measures to obtain chromatography high RST and oxidation
The concentration relationship curve (standard curve) of trimethylamine is as shown in Figure 2.Oxygen in human serum can be obtained using standard curve calculating
Change trimethylamine content.
Embodiment 2
In 4ml ml headspace bottle, 5.83 μm of ol/L isopropylamine 0.5ml, 100 μm of ol/L trimethylamine oxide 0.5ml, 20% 3 are added
Titanium chloride solution 0.1ml is then injected into 0.5ml saturation potassium hydroxide solution, continues after sealing water-bath 2 minutes in 80 DEG C of water
The water-bath 2 minutes in 80 DEG C of water, using head-space sampler from taken in ml headspace bottle headspace gas 1ml under 1 chromatographic condition of embodiment
Chromatographic isolation measurement is carried out, obtaining isopropylamine peak response value is 135244, and trimethylamine oxide peak response value is 4616, is calculated
Relative correction factor f=1.71.
Human serum sample containing internal standard compound (isopropylamine) carries out reaction, same color in ml headspace bottle under the above conditions
It is A that measurement, which obtains trimethylamine oxide peak response value, under spectral conditioni, internal standard compound (isopropylamine) peak response value be As, utilize following public affairs
Formula calculates human serum sample's trimethylamine oxide content mi:
Wherein, relative correction factor f=1.71, msFor the content (unit: μm ol/L) of internal standard compound (isopropylamine).
Claims (6)
1. it is a kind of using in ml headspace bottle in reaction assay human serum trimethylamine oxide content method, which is characterized in that utilize
Reaction generates trimethylamine in ml headspace bottle, carries out gas Chromatographic Determination and obtains the content of trimethylamine oxide, comprising the following steps:
A, human serum is injected into ml headspace bottle, reducing agent and vaporization agent is added and heated, front three will be aoxidized in human serum
Amine is reduced to trimethylamine and vaporizes；
B, the gas for taking certain volume from ml headspace bottle using head-space sampler, through gas-chromatography separation, fid detector measurement three
The content of trimethylamine oxide is calculated in methylamine.
2. it is according to claim 1 it is a kind of using in ml headspace bottle in reaction assay human serum trimethylamine oxide content side
Method, it is characterised in that: step A, the amount that human serum is added are the 1/8-1/4 of ml headspace bottle volume.
3. it is according to claim 1 it is a kind of using in ml headspace bottle in reaction assay human serum trimethylamine oxide content side
Method, it is characterised in that: step A, the reducing agent are titanium trichloride, and vaporization agent is to be saturated potassium hydroxide solution, ml headspace bottle volume:
Reducing agent volume: vaporization agent volume is 4:0.05~0.12:0.1~0.5.
4. it is according to claim 1 it is a kind of using in ml headspace bottle in reaction assay human serum trimethylamine oxide content side
Method, it is characterised in that: step A, the ml headspace bottle reaction temperature are 60~95 DEG C, and the reaction time is greater than 2min.
5. it is according to claim 1 it is a kind of using in ml headspace bottle in reaction assay human serum trimethylamine oxide content side
Method, it is characterised in that: step B measures trimethylamine oxide using quantified by external standard method.
6. it is according to claim 1 it is a kind of using in ml headspace bottle in reaction assay human serum trimethylamine oxide content side
Method, it is characterised in that: step B measures trimethylamine oxide using inner mark method ration, wherein internal standard compound is isopropylamine.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110894257A (en) * | 2019-12-06 | 2020-03-20 | 吉林大学 | Preparation method of trimethylamine oxide molecularly imprinted polymer |
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| CN103163084A (en) * | 2013-03-14 | 2013-06-19 | 广西工学院 | Characterization method for detecting freshness of shrimp product |
| CN105738539A (en) * | 2016-04-14 | 2016-07-06 | 中国农业科学院饲料研究所 | Method for determining content of trimethylamine in egg yolk through headspace gas chromatography |
| CN107340341A (en) * | 2017-06-27 | 2017-11-10 | 长沙都正生物科技有限责任公司 | A kind of TMAO immue quantitative detection reagent box and method |
| CN109870526A (en) * | 2017-12-01 | 2019-06-11 | 青岛谱尼测试有限公司 | A method of trimethylamine content in air is measured by Headspace-Gas Chromatography Analysis |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103163084A (en) * | 2013-03-14 | 2013-06-19 | 广西工学院 | Characterization method for detecting freshness of shrimp product |
| CN105738539A (en) * | 2016-04-14 | 2016-07-06 | 中国农业科学院饲料研究所 | Method for determining content of trimethylamine in egg yolk through headspace gas chromatography |
| CN107340341A (en) * | 2017-06-27 | 2017-11-10 | 长沙都正生物科技有限责任公司 | A kind of TMAO immue quantitative detection reagent box and method |
| CN109870526A (en) * | 2017-12-01 | 2019-06-11 | 青岛谱尼测试有限公司 | A method of trimethylamine content in air is measured by Headspace-Gas Chromatography Analysis |
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| A. Q. ZHANG ET AL.: "Determination of trimethylamine and related aliphatic amines in human urine by head-space gas chromatography", 《JOURNAL OF CHROMATOGRAPHY》 * |
| 冯忆 等: "环境中三甲胺的气相色谱测定", 《环境监测管理与技术》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110894257A (en) * | 2019-12-06 | 2020-03-20 | 吉林大学 | Preparation method of trimethylamine oxide molecularly imprinted polymer |
| CN110894257B (en) * | 2019-12-06 | 2023-06-02 | 吉林大学 | Preparation method of trimethylamine oxide molecularly imprinted polymer |
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Application publication date: 20191122 |