CN103698451A - Method for detecting amino acid in fermenting liquid by using gas chromatography-mass spectrometry - Google Patents
Method for detecting amino acid in fermenting liquid by using gas chromatography-mass spectrometry Download PDFInfo
- Publication number
- CN103698451A CN103698451A CN201310648730.8A CN201310648730A CN103698451A CN 103698451 A CN103698451 A CN 103698451A CN 201310648730 A CN201310648730 A CN 201310648730A CN 103698451 A CN103698451 A CN 103698451A
- Authority
- CN
- China
- Prior art keywords
- amino acid
- sample
- gas chromatography
- mass spectrum
- supernatant
- 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.)
- Pending
Links
Abstract
The invention relates to a method of detecting amino acid in a fermenting liquid by using gas chromatography-mass spectrometry, belonging to the technical field of biochemistry test measurement. The detection method comprises the following steps: (1) taking the fermenting liquid, centrifuging at a high speed, and respectively collecting cells and extracellular fluid; (2) preparing a fermenting liquid sample: adding acetonitrile into the taken supernatant liquor obtained in the step (1) to remove protein, performing vortex oscillation, centrifuging to collect a supernatant liquor III, adding the supernatant liquor III into an internal standard substance ribitol solution, and performing vacuum drying at room temperature to obtain an extracellular fluid sample II; (3) deriving the sample, adding a sugar derivation agent and an amino acid derivation agent; (4) performing gas chromatography-mass spectrometry analysis and data collection. The method has the advantages of simple sample treatment, simple operation, short amino acid detection time, high flexibility, high detection result repeatability, and realizability of preparation of multiple samples.
Description
Technical field
The invention belongs to biochemical test determination techniques field, particularly relate to a kind of gas chromatography-mass spectrum detect in fermentation liquor in amino acid whose method.
Background technology
Utilizing gas chromatography-mass spectrum to detect aspect amino acid metabolite, at present existing correlation technique report, as patent 201210046953.2, announced a kind of method that gas chromatography-mass spectrum detects urine amino acid, comprise the steps: that (1) completes urine enzyme to urine specimen to be checked and processes; (2) add interior mark product, adopt frozen ethanol to carry out the processing of protein precipitation, sample is dried up; (3) above-mentioned sample is carried out to methyl-monosilane derivation process; (4) adopt the step process standard amino acid of above-mentioned 1-3, obtain standard amino acid sample; (5) adopt gas chromatograph-mass spectrometer (GCMS) to detect standard amino acid sample and urine specimen to be checked; (6) using the testing result of standard amino acid sample as reference curve, with conventional algorithm, the amino acid of urine specimen to be checked is carried out quantitatively.This patented invention main application fields is urine examination, and this detection method is only confined to the detection of amino acid metabolite.
Detection research for metabolin in extracellular microbial, outside born of the same parents, can disclose microorganism metabolic rule under fermentation conditions, especially fermentation condition for the rule that affects of microorganism target product.The output of grasping the promotion target product that the metabolic rule of microorganism can be not only favourable promotes, and can determine that endobacillary metabolic flux distributes, and then utilizing genetic engineering means to realize the optimization and reconstruction of bacterium passway of metabolism, realize target product high-performance bio is synthetic.
For the metabolite analysis detection method limitation in fermentation liquor, be used for specific certain class material at present, can not realize multiclass metabolin and detect simultaneously.Because each component content complexity in fermentation liquor is various, disposal route for sample is very crucial, at present also in relevant fermentation liquor, the system of extracellular metabolin is processed detection method, make to develop a kind of more sensitive, wide spectrum, general fermentating metabolism object detecting method, identify the compound structure of various spectrums peak mapping, and with the integration of other dummy model, become the focus of microbial metabolism group research.
Summary of the invention
The object of this invention is to provide a kind of gas chromatography-mass spectrum and detect amino acid whose method in fermentation liquor, make up the blank to metabolin system detecting method in microorganism fermentation at present, overcome the shortcomings such as existing detection technique disturbing factor is a lot, complex operation, testing cost is high, sensitivity is low, sensing range is narrow.
The solution of the present invention is by such realization: a kind of gas chromatography-mass spectrum detects amino acid whose method in fermentation liquor, and detection method step comprises:
(1) get fermentation liquor, high speed centrifugation, collects respectively thalline and supernatant I;
(2) fermentation broth sample preparation: get the supernatant I that 60 ~ 350ul step 1) obtains, in clear liquid I, add acetonitrile to remove deproteinized, supernatant I and acetonitrile volume ratio are 1:0.5 ~ 1.5, vortex vibration, centrifugal collection supernatant III, adds internal standard compound ribitol solution again, and supernatant III volume and internal standard compound ribitol part by weight are 50 ~ 300ul:20 μ g, room temperature vacuum drying, obtains extracellular fluid sample II;
(3) in sample II analyte derivative: in step 2) obtaining, add respectively 50 ~ 150ul sugar derivating agent, constant temperature is placed 1.5 ~ 4h, add respectively more amino acid derived dose of 50 ~ 150ul, constant temperature is placed and is spent the night, derivative complete, sample II after centrifugal deriving, collects supernatant and obtains sample II in born of the same parents to be measured;
(4) utilize sample II in the born of the same parents to be measured that gas chromatography-mass spectrum obtains step 3), analyze and data acquisition.
As further restriction of the present invention, its sampling amount of described thalline reaches 0.2 ~ 1.0g/ml for dissolve artifact amount dry weight with cold methanol; Described cold methanol is for being chilled in advance-40 ℃ through cryostat groove; Described low-temperature extraction extracts 2 ~ 7h at-40 ℃ to-50 ℃.
As a further improvement on the present invention, described sugared derivating agent is that 0.1 ~ 0.3mg methoxamine hydrochloric acid is dissolved in 10ml pyridine solution preparation and obtains; Described amino acid derived dose is that 50 ~ 200ul trimethyl chlorosilane is dissolved in the trifluoroacetamide of 10ml.
As a further improvement on the present invention, its instrumental analysis condition of described gas chromatography-mass spectrum is: gas chromatography: chromatographic column is HP-5MS or the DB-5MS capillary column of 30 m * 0.25 mm, 300 ℃ of injector temperatures, 250 ℃ of detector temperatures, column temperature rise program equilibration time 3 min → 80 ℃ maintain 1min → 2 ℃/min and are warming up to 100 ℃ → 15 ℃/min and are warming up to 220 ℃ → 30 ℃/min and are warming up to 300 ℃ → 300 ℃ and maintain 3 min, sampling volume 1 μ L; Mass spectrum: ion gun EI, detecting device is level Four bar, ionization energy 70 eV, 280 ℃ of ion gun surface temperatures.
As a further improvement on the present invention, the method is applied to detect the amino acid in the fermentation liquor in the biological fermentation process of saccharomycete, lactic acid bacteria, clostridium, mould.
As a further improvement on the present invention, it is characterized in that, described amino acid is glutamic acid, glycocoll, halfcystine, serine, threonine, leucine, isoleucine, phenylalanine, valine, lysine, alanine, histidine, methionine, arginine, glutamine, asparatate, asparagine, tyrosine, proline.
Substantive distinguishing features of the present invention and marked improvement are: (1) the method can detect the detection of amino acids material in fermentation liquor, do not need fermentation liquor to carry out repeatedly complex process, save time and simplify the operation step, obtain in time microbial metabolism rule in sweat, analyze born of the same parents' intracellular metabolite flow.(2) the method adopts metabolin method in cold methanol extractive fermentation liquid, and its effect of extracting is good, obtains more metabolites kinds in fermentation liquor, is conducive to the analysis of metabolic rule.(3) the chromatographic peak analytical effect that sample analysis obtains is good, can the amino acid such as glutamic acid, glycocoll, halfcystine, serine, threonine, leucine, isoleucine, phenylalanine, valine, lysine, alanine, histidine, methionine, arginine, glutamine, asparatate, asparagine, tyrosine, proline be realized and being detected.
Embodiment
To the inventive method, further describe by embodiment below, these descriptions are not that content of the present invention is further limited.
In following examples, centrifugal condition is: centrifugal 10 min of 10000 rpm at-4 ℃.
In following examples, its instrumental analysis condition of gas chromatography-mass spectrum is: gas chromatography: chromatographic column is HP-5MS or the DB-5MS capillary column of 30 m * 0.25 mm, 300 ℃ of injector temperatures, 250 ℃ of detector temperatures, column temperature rise program equilibration time 3 min → 80 ℃ maintain 1min → 2 ℃/min and are warming up to 100 ℃ → 15 ℃/min and are warming up to 220 ℃ → 30 ℃/min and are warming up to 300 ℃ → 300 ℃ and maintain 3 min, sampling volume 1 μ L; Mass spectrum: ion gun EI, detecting device is level Four bar, ionization energy 70 eV, 280 ℃ of ion gun surface temperatures.
Following examples all can realize the amino acid such as glutamic acid, glycocoll, halfcystine, serine, threonine, leucine, isoleucine, phenylalanine, valine, lysine, alanine, histidine, methionine, arginine, glutamine, asparatate, asparagine, tyrosine, proline are detected.
Embodiment 1
Get saccharomycetes to make fermentation liquid.
(1) endocellular metabolism matter sample preparation: get 5ml fermentation liquor, the centrifugal thalline that obtains, with physiological saline, clean thalline 2 times,-40 ℃ of cold methanols dissolve artifact amount dry weight and reach 0.2g/ml, ultrasonication is to lysis, obtain lysate, low-temperature extraction lysate, cryogenic conditions is: at-40 ℃ to-50 ℃, extract 4h, cracking supernatant I after centrifugal collection 150ul extraction, adds internal standard compound ribitol solution, and cracking supernatant I volume and internal standard compound ribitol part by weight are 150ul:20 μ g, room temperature vacuum drying, obtains endocellular metabolism matter sample I.
(2) extracellular fluid sample preparation: get fermentation liquor, the centrifugal supernatant II that obtains, get 300ul supernatant II, in clear liquid II, add acetonitrile to remove deproteinized, supernatant II and acetonitrile volume ratio are 1:1.2, vortex vibration, the centrifugal supernatant III of collecting to obtain, adds internal standard compound ribitol solution again, and supernatant III volume and internal standard compound ribitol part by weight are 100ul:20 μ g, room temperature vacuum drying, obtains extracellular fluid sample II.
(3) in the sample I and sample II analyte derivative: in step 1) and step 2) obtaining, add respectively 100ul sugar derivating agent (sugared derivating agent be 0.1mg methoxamine hydrochloric acid be dissolved in preparation in 10ml pyridine solution obtain), constant temperature is placed 1.5h, add respectively more amino acid derived dose of 100ul (prepare and obtain for 100ul trimethyl chlorosilane is dissolved in the trifluoroacetamide of 10ml for amino acid derived dose), constant temperature is placed and is spent the night, derivative complete, distinguish sample I and sample II after centrifugal deriving, correspondence is collected supernatant separately and is obtained sample I and the outer sample II of born of the same parents to be measured in born of the same parents to be measured.
(4), according to its instrumental analysis condition of gas chromatography-mass spectrum, sample I in born of the same parents to be measured and the outer sample II of born of the same parents to be measured are analyzed and data acquisition.
Embodiment 2
Get mold fermentation liquid.
(1) endocellular metabolism matter sample preparation: get 5ml fermentation liquor, the centrifugal thalline that obtains, with physiological saline, clean thalline 3 times,-40 ℃ of cold methanols dissolve artifact amount dry weight and reach 1.0g/ml, ultrasonication is to lysis, obtain lysate, low-temperature extraction lysate, cryogenic conditions is: at-40 ℃ to-50 ℃, extract 5h, cracking supernatant I after centrifugal collection 100ul extraction, adds internal standard compound ribitol solution, and cracking supernatant I volume and internal standard compound ribitol part by weight are 100ul:20 μ g, room temperature vacuum drying, obtains endocellular metabolism matter sample I.
(2) extracellular fluid sample preparation: get fermentation liquor, the centrifugal supernatant II that obtains, get 350ul supernatant II, in clear liquid II, add acetonitrile to remove deproteinized, supernatant II and acetonitrile volume ratio are 1:0.7, vortex vibration, the centrifugal supernatant III of collecting to obtain, adds internal standard compound ribitol solution again, and supernatant III volume and internal standard compound ribitol part by weight are 200ul:20 μ g, room temperature vacuum drying, obtains extracellular fluid sample II.
(3) in the sample I and sample II analyte derivative: in step 1) and step 2) obtaining, add respectively 50ul sugar derivating agent (sugared derivating agent be 0.3mg methoxamine hydrochloric acid be dissolved in preparation in 10ml pyridine solution obtain), constant temperature is placed 2.0h, add respectively more amino acid derived dose of 150ul (prepare and obtain for 150ul trimethyl chlorosilane is dissolved in the trifluoroacetamide of 10ml for amino acid derived dose), constant temperature is placed and is spent the night, derivative complete, distinguish sample I and sample II after centrifugal deriving, correspondence is collected supernatant separately and is obtained sample I and the outer sample II of born of the same parents to be measured in born of the same parents to be measured.
(4), according to its instrumental analysis condition of gas chromatography-mass spectrum, sample I in born of the same parents to be measured and the outer sample II of born of the same parents to be measured are analyzed and data acquisition.
Embodiment 3
Get saccharomycetes to make fermentation liquid.
(1) endocellular metabolism matter sample preparation: get 5ml fermentation liquor, the centrifugal thalline that obtains, with physiological saline, clean thalline 3 times,-40 ℃ of cold methanols dissolve artifact amount dry weight and reach 0.6g/ml, ultrasonication is to lysis, obtain lysate, low-temperature extraction lysate, cryogenic conditions is: at-40 ℃ to-50 ℃, extract 3h, cracking supernatant I after centrifugal collection 200ul extraction, adds internal standard compound ribitol solution, and cracking supernatant I volume and internal standard compound ribitol part by weight are 200ul:20 μ g, room temperature vacuum drying, obtains endocellular metabolism matter sample I.
(2) extracellular fluid sample preparation: get fermentation liquor, the centrifugal supernatant II that obtains, get 60ul supernatant II, in clear liquid II, add acetonitrile to remove deproteinized, supernatant II and acetonitrile volume ratio are 1:1.5, vortex vibration, the centrifugal supernatant III of collecting to obtain, adds internal standard compound ribitol solution again, and supernatant III volume and internal standard compound ribitol part by weight are 50ul:20 μ g, room temperature vacuum drying, obtains extracellular fluid sample II.
(3) in the sample I and sample II analyte derivative: in step 1) and step 2) obtaining, add respectively 150ul sugar derivating agent (sugared derivating agent be 0.15mg methoxamine hydrochloric acid be dissolved in preparation in 10ml pyridine solution obtain), constant temperature is placed 2.5h, add respectively more amino acid derived dose of 50ul (prepare and obtain for 200ul trimethyl chlorosilane is dissolved in the trifluoroacetamide of 10ml for amino acid derived dose), constant temperature is placed and is spent the night, derivative complete, distinguish sample I and sample II after centrifugal deriving, correspondence is collected supernatant separately and is obtained sample I and the outer sample II of born of the same parents to be measured in born of the same parents to be measured.
(4), according to its instrumental analysis condition of gas chromatography-mass spectrum, sample I in born of the same parents to be measured and the outer sample II of born of the same parents to be measured are analyzed and data acquisition.
Embodiment 4
Extracting lactic acid fermented liquid.
(1) endocellular metabolism matter sample preparation: get 5ml fermentation liquor, the centrifugal thalline that obtains, with physiological saline, clean thalline 2 times,-40 ℃ of cold methanols dissolve artifact amount dry weight and reach 0.8g/ml, ultrasonication is to lysis, obtain lysate, low-temperature extraction lysate, cryogenic conditions is: at-40 ℃ to-50 ℃, extract 2h, cracking supernatant I after centrifugal collection 50ul extraction, adds internal standard compound ribitol solution, and cracking supernatant I volume and internal standard compound ribitol part by weight are 50ul:20 μ g, room temperature vacuum drying, obtains endocellular metabolism matter sample I.
(2) extracellular fluid sample preparation: get fermentation liquor, the centrifugal supernatant II that obtains, get 200ul supernatant II, in clear liquid II, add acetonitrile to remove deproteinized, supernatant II and acetonitrile volume ratio are 1:1.0, vortex vibration, the centrifugal supernatant III of collecting to obtain, adds internal standard compound ribitol solution again, and supernatant III volume and internal standard compound ribitol part by weight are 150ul:20 μ g, room temperature vacuum drying, obtains extracellular fluid sample II.
(3) in the sample I and sample II analyte derivative: in step 1) and step 2) obtaining, add respectively 120ul sugar derivating agent (sugared derivating agent be 0.2mg methoxamine hydrochloric acid be dissolved in preparation in 10ml pyridine solution obtain), constant temperature is placed 3.0h, add respectively more amino acid derived dose of 80ul (prepare and obtain for 50ul trimethyl chlorosilane is dissolved in the trifluoroacetamide of 10ml for amino acid derived dose), constant temperature is placed and is spent the night, derivative complete, distinguish sample I and sample II after centrifugal deriving, correspondence is collected supernatant separately and is obtained sample I and the outer sample II of born of the same parents to be measured in born of the same parents to be measured.
(4), according to its instrumental analysis condition of gas chromatography-mass spectrum, sample I in born of the same parents to be measured and the outer sample II of born of the same parents to be measured are analyzed and data acquisition.
Embodiment 5
Get saccharomycetes to make fermentation liquid.
(1) endocellular metabolism matter sample preparation: get 5ml fermentation liquor, the centrifugal thalline that obtains, with physiological saline, clean thalline 3 times,-40 ℃ of cold methanols dissolve artifact amount dry weight and reach 0.5g/ml, ultrasonication is to lysis, obtain lysate, low-temperature extraction lysate, cryogenic conditions is: at-40 ℃ to-50 ℃, extract 3h, cracking supernatant I after centrifugal collection 200ul extraction, adds internal standard compound ribitol solution, and cracking supernatant I volume and internal standard compound ribitol part by weight are 150ul:20 μ g, room temperature vacuum drying, obtains endocellular metabolism matter sample I.
(2) extracellular fluid sample preparation: get fermentation liquor, the centrifugal supernatant II that obtains, get 300ul supernatant II, in clear liquid II, add acetonitrile to remove deproteinized, supernatant II and acetonitrile volume ratio are 1:0.5, vortex vibration, the centrifugal supernatant III of collecting to obtain, adds internal standard compound ribitol solution again, and supernatant III volume and internal standard compound ribitol part by weight are 300ul:20 μ g, room temperature vacuum drying, obtains extracellular fluid sample II.
(3) in the sample I and sample II analyte derivative: in step 1) and step 2) obtaining, add respectively 100ul sugar derivating agent (sugared derivating agent be 0.2mg methoxamine hydrochloric acid be dissolved in preparation in 10ml pyridine solution obtain), constant temperature is placed 4.0h, add respectively more amino acid derived dose of 120ul (prepare and obtain for 120ul trimethyl chlorosilane is dissolved in the trifluoroacetamide of 10ml for amino acid derived dose), constant temperature is placed and is spent the night, derivative complete, distinguish sample I and sample II after centrifugal deriving, correspondence is collected supernatant separately and is obtained sample I and the outer sample II of born of the same parents to be measured in born of the same parents to be measured.
(4), according to its instrumental analysis condition of gas chromatography-mass spectrum, sample I in born of the same parents to be measured and the outer sample II of born of the same parents to be measured are analyzed and data acquisition.
Claims (7)
1. gas chromatography-mass spectrum detects an amino acid whose method in fermentation liquor, it is characterized in that, detection method step comprises:
(1) get fermentation liquor, high speed centrifugation, collects respectively thalline and supernatant I;
(2) fermentation broth sample preparation: get the supernatant I that 60 ~ 350ul step 1) obtains, in clear liquid I, add acetonitrile to remove deproteinized, supernatant I and acetonitrile volume ratio are 1:0.5 ~ 1.5, vortex vibration, centrifugal collection supernatant III, adds internal standard compound ribitol solution again, and supernatant III volume and internal standard compound ribitol part by weight are 50 ~ 300ul:20 μ g, room temperature vacuum drying, obtains extracellular fluid sample II;
(3) in sample II analyte derivative: in step 2) obtaining, add respectively 50 ~ 150ul sugar derivating agent, constant temperature is placed 1.5 ~ 4h, add respectively more amino acid derived dose of 50 ~ 150ul, constant temperature is placed and is spent the night, derivative complete, sample II after centrifugal deriving, collects supernatant and obtains sample II in born of the same parents to be measured;
(4) utilize sample II in the born of the same parents to be measured that gas chromatography-mass spectrum obtains step 3), analyze and data acquisition.
2. a kind of gas chromatography-mass spectrum according to claim 1 detects amino acid whose method in fermentation liquor, it is characterized in that, its sampling amount of described thalline reaches 0.2 ~ 1.0g/ml for dissolve artifact amount dry weight with cold methanol; Described cold methanol is for being chilled in advance-40 ℃ through cryostat groove; Described low-temperature extraction extracts 2 ~ 7h at-40 ℃ to-50 ℃.
3. a kind of gas chromatography-mass spectrum according to claim 1 and 2 detects amino acid whose method in fermentation liquor, it is characterized in that, described sugared derivating agent is that 0.1 ~ 0.3mg methoxamine hydrochloric acid is dissolved in preparation in 10ml pyridine solution and obtains; Described amino acid derived dose is that 50 ~ 200ul trimethyl chlorosilane is dissolved in the trifluoroacetamide of 10ml.
4. a kind of gas chromatography-mass spectrum according to claim 3 detects amino acid whose method in fermentation liquor, it is characterized in that, its instrumental analysis condition of described gas chromatography-mass spectrum is: gas chromatography: chromatographic column is HP-5MS or the DB-5MS capillary column of 30 m * 0.25 mm, 300 ℃ of injector temperatures, 250 ℃ of detector temperatures, column temperature rise program equilibration time 3 min → 80 ℃ maintain 1min → 2 ℃/min and are warming up to 100 ℃ → 15 ℃/min and are warming up to 220 ℃ → 30 ℃/min and are warming up to 300 ℃ → 300 ℃ and maintain 3 min, sampling volume 1 μ L; Mass spectrum: ion gun EI, detecting device is level Four bar, ionization energy 70 eV, 280 ℃ of ion gun surface temperatures.
5. a kind of gas chromatography-mass spectrum detects amino acid whose method in fermentation liquor according to claim 4, it is characterized in that, the method is applied to detect the amino acid in the fermentation liquor in the biological fermentation process of saccharomycete, lactic acid bacteria, clostridium, mould.
6. according to a kind of gas chromatography-mass spectrum described in claim 1 ~ 2 or 4 ~ 5 any one, detect amino acid whose method in fermentation liquor, it is characterized in that, described amino acid is glutamic acid, glycocoll, halfcystine, serine, threonine, leucine, isoleucine, phenylalanine, valine, lysine, alanine, histidine, methionine, arginine, glutamine, asparatate, asparagine, tyrosine, proline.
7. a kind of gas chromatography-mass spectrum according to claim 3 detects amino acid whose method in fermentation liquor, it is characterized in that, described amino acid is glutamic acid, glycocoll, halfcystine, serine, threonine, leucine, isoleucine, phenylalanine, valine, lysine, alanine, histidine, methionine, arginine, glutamine, asparatate, asparagine, tyrosine, proline.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310648730.8A CN103698451A (en) | 2013-12-05 | 2013-12-05 | Method for detecting amino acid in fermenting liquid by using gas chromatography-mass spectrometry |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310648730.8A CN103698451A (en) | 2013-12-05 | 2013-12-05 | Method for detecting amino acid in fermenting liquid by using gas chromatography-mass spectrometry |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103698451A true CN103698451A (en) | 2014-04-02 |
Family
ID=50360050
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310648730.8A Pending CN103698451A (en) | 2013-12-05 | 2013-12-05 | Method for detecting amino acid in fermenting liquid by using gas chromatography-mass spectrometry |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103698451A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104407093A (en) * | 2014-11-24 | 2015-03-11 | 河南巨龙生物工程股份有限公司 | Method for quickly detecting content of L-glutamine in fermentation liquor |
| CN109298100A (en) * | 2018-11-28 | 2019-02-01 | 长春博瑞农牧集团股份有限公司 | A kind of metabonomic analysis methods of Cultures of S. cerevisiae |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0452554A (en) * | 1990-06-20 | 1992-02-20 | Ichiro Yoshihara | Easy and quick gas chromatographic analyzing method of cyclic amino acid amine, serotonin and cholesterol in serum |
| KR20020084610A (en) * | 2001-05-03 | 2002-11-09 | 재단법인 서울의과학연구소 | Simultaneous screening for multiple organic acids, amino acids and glycines using GC/MS |
| CN102621248A (en) * | 2012-04-18 | 2012-08-01 | 长沙高新开发区鷁巢生物科技有限公司 | Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with one-step derivation method |
| CN102621247A (en) * | 2012-04-18 | 2012-08-01 | 王益超 | Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with two-step derivation method |
-
2013
- 2013-12-05 CN CN201310648730.8A patent/CN103698451A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0452554A (en) * | 1990-06-20 | 1992-02-20 | Ichiro Yoshihara | Easy and quick gas chromatographic analyzing method of cyclic amino acid amine, serotonin and cholesterol in serum |
| KR20020084610A (en) * | 2001-05-03 | 2002-11-09 | 재단법인 서울의과학연구소 | Simultaneous screening for multiple organic acids, amino acids and glycines using GC/MS |
| CN102621248A (en) * | 2012-04-18 | 2012-08-01 | 长沙高新开发区鷁巢生物科技有限公司 | Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with one-step derivation method |
| CN102621247A (en) * | 2012-04-18 | 2012-08-01 | 王益超 | Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with two-step derivation method |
Non-Patent Citations (4)
| Title |
|---|
| INES STARKE ET AL: "Separation, identification, and quantification of amino acids in L-lysine fermentation potato juices by gas chromatography–mass spectrometry", 《FRESENIUS J ANAL CHEM》 * |
| 周慧 等: "柱前衍生化ＨＰＬＣ法测定发酵液中Ｌ-瓜氨酸和Ｌ-鸟氨酸含量", 《南京工业大学学报（自然科学版）》 * |
| 孙茂成: "保加利亚乳杆菌代谢组学样品的前处理研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
| 徐正华 等: "气相色谱-质谱法同时测定乳酸和7种糖的研究", 《分析科学学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104407093A (en) * | 2014-11-24 | 2015-03-11 | 河南巨龙生物工程股份有限公司 | Method for quickly detecting content of L-glutamine in fermentation liquor |
| CN104407093B (en) * | 2014-11-24 | 2016-01-20 | 河南巨龙生物工程股份有限公司 | A kind of method of Glu content in quick detection fermentation liquor |
| CN109298100A (en) * | 2018-11-28 | 2019-02-01 | 长春博瑞农牧集团股份有限公司 | A kind of metabonomic analysis methods of Cultures of S. cerevisiae |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109298115B (en) | Quantitative detection method for multiple metabolites in biological sample and metabolic chip | |
| CN101430307B (en) | Method for simultaneously analyzing amino acid and organic acid metabolite spectrum | |
| US20160002696A1 (en) | Method to identify bacterial species by means of gas chromatography/mass spectrometry in biological samples | |
| CN102091118B (en) | Quality detection method for rhodiola crenulata | |
| CN104391060B (en) | The sample pre-treatments of GC-MS research the pathogen of Botrytis cinerea metabolism group and detection method | |
| CN107085066B (en) | A kind of pre-treating method for LC-MS detection Aspergillus flavus metabolism group | |
| CN103293245A (en) | GC-MS (Gas Chromatography-Mass Spectrometer)/MS (Mass Spectrometer) method for simultaneously analyzing four tobacco-specific nitrosamines (TSNAs) in main stream smoke of cigarette | |
| CN103257194A (en) | GC-MS/MS (Gas chromatography - tandem mass spectrometry) method for simultaneously analyzing three polycyclic aromatic hydrocarbons and four tobacco-specific nitrosamines in main stream smoke of cigarettes | |
| CN102944636B (en) | High-efficiency liquid chromatography to mass spectrum detection method for ethyl carbamate in distilled liquor | |
| CN103675128A (en) | Method for detecting amino acid in intracellular and extracellular fluids by gas chromatography-mass spectrum | |
| CN103792312A (en) | Method for detecting amino acids and saccharides in fermentation liquor through gas chromatography-mass spectrometry | |
| CN103364510A (en) | Method for evaluating cytotoxicity of environmental pollutants by adopting HPLC-MS/MS (high performance liquid chromatography and mass spectrometry or mass spectrometry) and metabolic flux analysis system | |
| CN103698451A (en) | Method for detecting amino acid in fermenting liquid by using gas chromatography-mass spectrometry | |
| CN106153785A (en) | A kind of online sample introduction of aflatoxin analyzes method | |
| CN104849363A (en) | Cordate houttuynia wall-breaking decoction pieces fingerprinting construction and quality detection method thereof | |
| CN103808841B (en) | Gas chromatography-mass spectrum detects the method for organic acid in fermentation liquor, amino acid, sugar | |
| CN101315352B (en) | Method for measuring myriocin content in Chinese Caterpillar Fungus | |
| Deng et al. | Investigation of tomato plant defence response to tobacco mosaic virus by determination of methyl salicylate with SPME-capillary GC-MS | |
| CN103675132B (en) | Gas chromatography-mass spectrum detects middle amino acid, sugar, organic acid method in cell | |
| CN103675130A (en) | Gas chromatography-mass spectrometry detection method for amino acids and carbohydrates in extracellular fluid and intracellular fluid | |
| CN103675178A (en) | Gas chromatography-mass spectrometry detection method for organic acids and sugar in fermentation liquid | |
| CN103675133A (en) | Method for detecting amino acids and organic acids in cells through gas chromatography-mass spectrometry | |
| CN103698450A (en) | Method for detecting intracellular organic acid through gas chromatography-mass spectrometry | |
| CN103675180A (en) | Gas chromatography-mass spectrometry detection method for intracellular carbohydrates and amino acids | |
| CN103713074A (en) | Detection method for sugar in intracellular and extracellular fluids through gas chromatography-mass spectrometry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C05 | Deemed withdrawal (patent law before 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140402 |