CN105548407B - The detection method of modified nucleoside in urine - Google Patents
The detection method of modified nucleoside in urine Download PDFInfo
- Publication number
- CN105548407B CN105548407B CN201610038315.4A CN201610038315A CN105548407B CN 105548407 B CN105548407 B CN 105548407B CN 201610038315 A CN201610038315 A CN 201610038315A CN 105548407 B CN105548407 B CN 105548407B
- Authority
- CN
- China
- Prior art keywords
- urine
- modified nucleoside
- sample
- filter paper
- methanol
- 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.)
- Active
Links
Classifications
-
- 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
-
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Abstract
The invention provides a kind of detection method of modified nucleoside in urine, following steps are specifically included：Urine pretreatment after be stored on filter paper, after by the way that sample to be tested is obtained by extraction；The sample to be tested is detected by LC-MS, the species and content of modified nucleoside are obtained.In the detection method that the present invention is provided, by the way that urine is stored on filter paper again by extraction process, with LC-MS method, qualitative and quantitative detection quickly and efficiently can be carried out to the modified nucleoside in urine.Due to urine is stored on filter paper, urine sample is converted into solid form and preserved, stability is greatly improved, so as to avoid microorganism pollution from detecting sample.Moreover, the method that provides of the present invention is also more simpler than existing freezing, conveniently.
Description
Technical field
The present invention relates to liquid spectrum mass spectrometric hyphenated technique field, in particular to a kind of detection of modified nucleoside in urine
Method.
Background technology
Modified nucleoside is primarily present in RNA, be acted on after transcription by specific transmethylase and ligase it is many
Poly- nucleoside molecule and form.RNA metabolism is required for specific enzyme to be catalyzed, normal nucleotide enzymolysis after degradable or phosphatideization again
Utilize, and to be the glycosidic bond of normal nucleotide, base, sugared hydroxyl be modified or combine two-by-two the unique knot of generation to modified nucleoside
Structure, due to highly being modified, the enzymatic activity that degraded modified nucleoside needs is higher, high specificity, and therefore, modified nucleoside is from cell
Discharge, discharged eventually through urine.Therefore, the content of modified nucleoside can be used to evaluate the nucleoside metabolism shape of human body in urine
Condition.
At present, have been reported and modified nucleoside in urine is detected using capillary electrophoresis, but this method is detected
Modified nucleoside classification it is few.It is additionally, since in urine and contains multiple-microorganism.If preserved in liquid form for a long time, micro- life
Thing will pollute urine, and it is all that will be deposited after urine sampling to be detected to influence in the accuracy of testing result, existing detection method
Freezen protective in refrigerator is stored in, is thawed again during detection, refrigerating plant is not only needed, defrosting step also make it that detection time is elongated.
In view of this, it is special to propose the present invention.
The content of the invention
It is an object of the invention to provide a kind of detection method of modified nucleoside in urine.For detection sample easily by micro- life
Urine is stored on filter paper in the shortcoming of thing pollution, the detection method that the present invention is provided, not only stabilizing it property is carried significantly
It is high, it is to avoid the pollution of microorganism, but also with simple, convenient advantage.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
The detection method of modified nucleoside, comprises the following steps in urine：
(1) urine pretreatment after be stored on filter paper, after by the way that sample to be tested is obtained by extraction；
(2) sample to be tested is detected by LC-MS, obtains the species and content of modified nucleoside.
In the detection method that the present invention is provided, by the way that urine is stored on filter paper again by extraction process, with liquid
Matter method for combined use, quickly and efficiently can carry out qualitative and quantitative detection to the modified nucleoside in urine.
Due to urine is stored on filter paper, urine sample is converted into solid form and preserved, stability is significantly
Improve, so as to avoid microorganism pollution from detecting sample.Moreover, the method that the present invention is provided is also more simpler than existing freezing
Easily, conveniently.
Preferably, specifically included in step (1), the step of the extraction：Filter paper is put into methanol water mixed solution
Supernatant is taken as sample to be tested after ultrasound, centrifugation.
In the detection method that the present invention is provided, in order to which the urine specimen being stored on filter paper is fully reduced, by filter paper
Piece be put into methanol water mixed solution carry out ultrasound so that each component on filter paper is fully dissolved into solution, after pass through again
Centrifugation obtains supernatant, regard this supernatant as sample to be tested.
During actually detected, the filter paper for preserving urine can be punched, aperture is 3mm, obtains a diameter of 3mm's
Filter paper dick, takes appropriate filter paper dick to be placed in centrifuge tube, adds and ultrasound, centrifugally operated are carried out after methanol aqueous solution.
In order to which each component ensured in filter paper fully can be dissolved in solution, methanol in methanol aqueous solution used
Volume content be 5%-50%, ultrasonic time is 20-90min.
After ultrasound, modified nucleoside is not only contained in solution, also containing other solid impurities, the accurate of testing result is influenceed
Property, in order to which impurity is removed as much as possible, in the detection method that the present invention is provided, the rotating speed of centrifugation is controlled in 10000-
18000rpm, the time is 5-30min.
In order to ensure in the stability for detecting sample, step (1), the step on filter paper is stored in after the urine pretreatment
Suddenly specifically include：Add and dripped after the mixing of 8- bromines guanosines on filter paper into urine, and air-dry more than 3 hours.
In the above-mentioned methods, the 8- bromines guanosine of addition is as internal standard compound, to carry out quantitative analysis to modified nucleoside.
Preferably, step (2) is specifically included：
(21) standard items are weighed, are dissolved with methanol, the titer of various concentrations is obtained；
(22) weigh internal standard compound, be separately added into after being dissolved with methanol aqueous solution in the titer, after ultra-pure water constant volume from
The heart, takes supernatant as titer to be measured；
(23) sample to be tested and the titer to be measured are detected by LC-MS, obtains the species of modified nucleoside
And content.
In the detection method that the present invention is provided, the content of the modified nucleoside in urine is detected using internal standard method.
In detection process, weigh standard items is completely dissolved it with methanol, obtains the titer of various concentrations, into titer in addition
Mark thing after through processing obtain titer to be measured, to carry out LC-MS detection, respectively with the peak area of each modified nucleoside with it is interior
The concentration for marking each modified nucleoside in the comparison urine sample of thing peak area carries out linear regression, obtains the linear equation of each modified nucleoside,
For carrying out quantitative analysis to each modified nucleoside in urine.
In order to ensure the accuracy of LC-MS testing result, the present invention uses specific liquid-phase condition and Mass Spectrometry Conditions,
Specifically, in step (23), the liquid-phase condition of the LC-MS detection is：The flow velocity of mobile phase is 0.1-1.0mL/min；Enter
Sample amount 1-20 μ L；Bonded-phase chromatography post；20-40 DEG C of column temperature；A and B is methanol aqueous solution as mobile phase, wherein A, and B is formic acid
Methanol solution, gradient elution program is as shown in table 1.
The gradient elution program of table 1
Mass Spectrometry Conditions are：Using ESI ion guns, cation MRM scannings, atomization gas flow velocity 8-20L/min, gas curtain gas velocity
8-20L/min, collision gas flow velocity 5-15L/min, ion source voltage 2000-4000V, 200-400 DEG C of ion source temperature.
After above-mentioned liquid chromatogram and Mass Spectrometer Method, effectively the modified nucleoside in urine can be carried out qualitative and quantitative
Analysis.Also, analyze speed is fast, completing once analysis only needs 13 minutes.
Because modified nucleoside species is more, in order to guarantee as much as possible to separate the modified nucleoside in urine, further
Preferably, in the liquid-phase condition, mobile phase A is 0.1-10% methanol aqueous solutions, and B is 0.1-10% formic acid methanol solutions.
Based on same consideration, in the liquid-phase condition, the bonded-phase chromatography post is C18/C8 chromatographic columns, and specification is
(50-100) mm × (1.8-3) mm, (1.8-3) μm.
Preferably, in step (21), the standard items include pseudouridine, cytidine, M1A, urine
Pyrimidine nucleoside, adenosine, inosine, guanosine, 1-methyl hypoxanthine nucleosides, 5-methyl-uridin, Huang
Purine nucleosides, 3- methyluridines, M1G, 6- methyladenosines, M2G, N4- acetyl group cytidine and N2, N2- bis-
Methylguanosine.
In the detection method that provides of the present invention, standard items used include 16 kinds, and determining for 16 kinds of modified nucleosides can be achieved
Property and quantitative analysis so that detection range improve.
Preferably, in step (22), the internal standard compound is 8- bromine guanosines.
In the detection method that the present invention is provided, from 8- bromines guanosine as internal standard compound, good stability can be used in urine
Modified nucleoside in liquid carries out quantitative analysis.
Compared with prior art, beneficial effects of the present invention are：
(1) urine is stored on filter paper by the present invention, and stabilizing it property is greatly improved, it is to avoid microorganism is to detection sample
The pollution of product, solves the problem of urine can not for a long time be preserved, transported.
(2) present invention may be such that each component on filter paper is fully dissolved out by the optimization to extraction process, can be also
The authenticity of original detection sample so that the accuracy of testing result is greatly improved.
(3) present invention is realized to urine by the optimization to liquid-phase condition and Mass Spectrometry Conditions in LC-MS detection process
The qualitative and quantitative analysis of modified nucleoside in liquid, sensitivity is high, and accuracy is high, and analyze speed is fast.
(4) in the detection method that the present invention is provided, using 8- bromines guanosine as internal standard compound, using pseudouridine, born of the same parents
Pyrimidine nucleoside, M1A, uridine (U), adenosine, inosine, guanosine, 1- methyl
Xanthosine, 5-methyl-uridin, xanthosine, 3- methyluridines, M1G, 6- methyladenosines, M2G,
N4- acetyl group cytidine and N2, N2- dimethylguanosine this modified nucleoside, can be to 16 kinds of modifications in urine as standard items in 16
Nucleosides carries out qualitative and quantitative analysis simultaneously, meet it is actually detected the need for.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
The accompanying drawing to be used needed for having technology description is briefly described.
Fig. 1 is the detection spectrogram of the embodiment of the present invention.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment
Condition person, the condition advised according to normal condition or manufacturer is carried out.Agents useful for same or the unreceipted production firm person of instrument, are
The conventional products that can be obtained by commercially available purchase.
Embodiment
Instrument and material：
The tandem mass spectrometer of Agilent companies of the U.S. 6495, the liquid chromatographs of Angilent 1290, chromatographic column is Agilent
C18 posts, specification is 50mm × 3.0mm, 2.7 μm.
Medicine and reagent：
Standard items：Pseudouridine, cytidine, M1A, uridine, adenosine, secondary Huang
Purine nucleosides, guanosine, 1-methyl hypoxanthine nucleosides, 5-methyl-uridin, xanthosine, 3- methyluridines, 1- first
Base guanosine, 6- methyladenosines, M2G, N4- acetyl group cytidine, N2, N2- dimethylguanosines are bought from Sigma-
Aldrich.
Internal standard compound：8- bromines guanosine (8-Br-G), buys from Sigma-Aldrich companies.
Methanol：Chromatographically pure, Merck companies.
Formic acid：Chromatographically pure, Merck companies.
Ultra-pure water：Thermo companies.
Urine sample：Volunteer's urine sample.
Above-mentioned 16 kinds of standard items are accurately weighed, is dissolved respectively with methanol, 10mg/mL solution is configured to respectively, is remixed
10 μ g/mL hybrid standard product storing solution is into each standard concentration.
A certain amount of internal standard compound 8- bromine guanosines are accurately weighed, with 50% methanol water mixed solution (the i.e. volume of first alcohol and water
Than for 1:1) dissolve and the storing solution for 0.3mg/mL is diluted with water, add water the internal standard solution for being diluted to that content is 0.15mg/mL.
Take respectively the μ L of hybrid standard product storing solution 5,10 μ L, 20 μ L, 50 μ L, 100 μ L, 200 μ L, 400 μ L, 800 μ L in
In 1.5mL centrifuge tubes, and 20 μ L internal standard solutions are separately added into, 1mL is settled to respectively with ultra-pure water, be put into supercentrifuge and carry out
Centrifugally operated, rotating speed is set to 12000rpm, and set of time is 5min.Centrifugation takes supernatant as titer to be measured after terminating.
Take urine 100ul, and add 20ul internal standard solution, fully mix, after mixed liquor is dropped on filter paper, room temperature wind
It is dry more than 3 hours.
Punched on air-dried filter paper, aperture is 3mm, obtains a diameter of 3mm filter paper dick.Take 2 filter paper dicks
It is placed in 1.5mL centrifuge tubes, with 5% methanol water dissolves, (i.e. the volume ratio of first alcohol and water is 5:95), and ultrasound 20-30 minutes,
Place into afterwards and centrifugally operated is carried out in centrifuge, the rotating speed of centrifugation is controlled in 12000rpm, and the time is 20min.
Centrifugation takes supernatant as sample to be tested after terminating.
Obtained sample to be tested and titer sample introduction to be measured are detected into LC-MS instrument.
The liquid-phase condition of LC-MS detection is：Flow velocity is 0.1-1.0mL/min；Sample size 1-20 μ L；Column temperature 20-
40℃；A and B is as mobile phase, and wherein A is that (i.e. the volume ratio of first alcohol and water is 0.1 to 0.1% methanol aqueous solution:99.9), B is
(i.e. the volume ratio of formic acid and methanol is 0.1 to 0.1% formic acid methanol solution:99.9), gradient elution program is as shown in table 1.
Mass Spectrometry Conditions are：Using ESI ion guns, cation MRM scannings, atomization gas flow velocity 10L/min, gas curtain gas velocity
10L/min, collision gas flow velocity 8L/min, ion source voltage 2500V, 400 DEG C of ion source temperature.
Its testing result is as shown in Figure 1.In Fig. 1 each numeral represent the component that represents as：
1：Pseudouridine；2：Cytidine；3：M1A；4：Uridine；5：Adenosine；6：
Inosine；7：Guanosine；8：1-methyl hypoxanthine nucleosides；9：5-methyl-uridin；10：Xanthosine；11：
3- methyluridines；12：M1G；13：6- methyladenosines；14：M2G；15：N4- acetyl group cytidines；16：N2,
N2- dimethylguanosines；17：The bromo- guanosines of 8-.
From figure 1 it appears that each component peak sequence is：Pseudouridine, cytidine, M1A,
Uridine, adenosine, inosine, guanosine, 1-methyl hypoxanthine nucleosides, 5-methyl-uridin,
Xanthosine, 3- methyluridines, M1G, 6- methyladenosines, M2G, N4- acetyl group cytidine, N2, N2- bis-
Methylguanosine, 8- bromine guanosines.
By the peak area of each modified nucleoside detected in titer to be measured and the comparison work mark of the peak area of internal standard compound
The concentration of each modified nucleoside carries out linear regression in quasi- liquid, obtains the standard curve of each modified nucleoside.Its result is as shown in table 2.
The standard curve and the content in urine sample of 2 16 kinds of modified nucleosides of table
According to the ratio between the peak area of each modified nucleoside of urine to be measured and internal standard compound peak area, the standard substituted into respectively in table 1
Curve, obtains the content of each modified nucleoside in urine.
Detection method reappearance analyzes (rate of recovery test)
Take 200 μ L samples to be tested, be separately added into the blank water of 100 μ L hybrid standard product storing solutions and same volume in
In 1.5mL centrifuge tubes, then 20 μ L internal standard solutions are separately added into, are settled to 1mL respectively with ultra-pure water, are put into supercentrifuge
5min is centrifuged under 12000rpm rotating speed, it is sufficiently mixed.Centrifugation takes supernatant to carry out LC-MS detection after terminating.Liquid matter
It is combined the liquid-phase condition and Mass Spectrometry Conditions be the same as Example 1 of detection.
By the working curve in the ratio between the peak area of each modified nucleoside detected and internal standard compound peak area substitution table 2, obtain
To the content of each modified nucleoside.
Operation is repeated 3 times, as a result relative standard deviation 1.98%-3.67%, sample recovery rate is in 81%-98.7%, table
The favorable reproducibility of bright detection method.
Detection method precision accuracy analysis (day internal difference difference analysis in the daytime)
Under the conditions of the identical liquid chromatography mass of be the same as Example 1, the hybrid standard product storing solution of different volumes is taken to carry out
Detection, in 1 day in continuous sample introduction 3 times, 3 days daily sample introduction once, as a result in a few days CV is fluctuated in 1.98%-3.67%, in the daytime CV
In 3.51%-11.24% fluctuations, meet the requirements.
The detection method analyze speed that the present invention is provided it can be seen from the above is fast, and completing once analysis only needs 13
Minute.16 kinds of modified nucleosides in urine can be carried out qualitative and fixed by the detection method provided using the present invention faster and betterly
Amount analysis.
Although illustrate and describing the present invention with specific embodiment, but it will be appreciated that without departing substantially from the present invention's
Many other changes and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (1)
1. the detection method of modified nucleoside in urine, it is characterised in that comprise the following steps：
(1) urine pretreatment after be stored on filter paper, after by the way that sample to be tested is obtained by extraction；
(2) sample to be tested is detected by LC-MS, obtains the species and content of modified nucleoside；
Specifically included in step (1), the step of the extraction：Filter paper is put into methanol water mixed solution through ultrasound, centrifugation
After take supernatant as sample to be tested；
Step (2) is specifically included：
(21) standard items are weighed, are dissolved with methanol, the titer of various concentrations is obtained；
(22) internal standard compound is weighed, is separately added into after being dissolved with methanol aqueous solution in the titer, centrifuges, takes after ultra-pure water constant volume
Supernatant is used as titer to be measured；
(23) sample to be tested and the titer to be measured are detected by LC-MS, obtains the species of modified nucleoside and contain
Amount；
In step (23), the liquid-phase condition of the LC-MS detection is：Flow velocity is 0.1-1.0mL/min；Sample size 1-20 μ L；
Bonded-phase chromatography post；20-40 DEG C of column temperature；A and B is as mobile phase, and wherein A is methanol aqueous solution, and B is formic acid methanol solution, ladder
Degree elution program is 0-0.5min, A：97%th, B：3%；0.5-3min, A：97~5%, B：3~95%；3-3.01min, A：5-
97%th, B：95-3%；3.01-11min, A：97%th, B：3%；11.01min, A：97%th, B：3%；13min, A：97%th, B：
3%；
Mass Spectrometry Conditions are：Using ESI ion guns, cation MRM scannings, atomization gas flow velocity 8-20L/min, gas curtain gas velocity 8-
20L/min, collision gas flow velocity 5-15L/min, ion source voltage 2000-4000V, 200-400 DEG C of ion source temperature；
In step (21), the standard items include pseudouridine, cytidine, M1A, uridine, gland
Purine nucleosides, inosine, guanosine, 1-methyl hypoxanthine nucleosides, 5-methyl-uridin, xanthosine, 3-
Methyluridine, M1G, 6- methyladenosines, M2G, N4- acetyl group cytidine and N2, N2- dimethylguanosine；
In step (22), the internal standard compound is 8- bromine guanosines；
In the liquid-phase condition, mobile phase A is 0.1-10% methanol aqueous solutions, and B is 0.1-10% formic acid methanol solutions；
In the liquid-phase condition, the bonded-phase chromatography post is C18 or C8 chromatographic columns, and specification is (50-100) mm × (1.8-3)
Mm, (1.8-3) μm；
In step (1), the rotating speed of the centrifugation is 10000-18000rpm, and the time is 5-20min；
In step (1), the step being stored in after the urine pretreatment on filter paper is specifically included：8- bromine birds are added into urine
Drip on filter paper, and air-dry more than 3 hours after glycosides mixing；
The modified nucleoside include pseudouridine, cytidine, M1A, uridine, adenosine,
Inosine, guanosine, 1-methyl hypoxanthine nucleosides, 5-methyl-uridin, xanthosine, 3- methyluridines,
M1G, 6- methyladenosines, M2G, N4- acetyl group cytidine and N2, N2- dimethylguanosine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610038315.4A CN105548407B (en) | 2016-01-20 | 2016-01-20 | The detection method of modified nucleoside in urine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610038315.4A CN105548407B (en) | 2016-01-20 | 2016-01-20 | The detection method of modified nucleoside in urine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105548407A CN105548407A (en) | 2016-05-04 |
| CN105548407B true CN105548407B (en) | 2017-07-14 |
Family
ID=55827740
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610038315.4A Active CN105548407B (en) | 2016-01-20 | 2016-01-20 | The detection method of modified nucleoside in urine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105548407B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108037220A (en) * | 2018-01-26 | 2018-05-15 | 上海上药第生化药业有限公司 | A kind of separation method of nucleosides and alkali radical species and its application |
| CN108344815A (en) * | 2018-01-26 | 2018-07-31 | 上海上药第生化药业有限公司 | A kind of separation method of base substance and its application |
| CN112858499A (en) * | 2020-12-31 | 2021-05-28 | 郑州大学第一附属医院 | Method for simultaneously determining modified nucleoside and creatinine in urine |
| CN112834644A (en) * | 2020-12-31 | 2021-05-25 | 郑州大学第一附属医院 | Bladder cancer related combined marker and detection kit |
| CN115487542A (en) * | 2022-09-02 | 2022-12-20 | 浙江大学 | Solid-phase extraction method for modified nucleosides in urine and application of solid-phase extraction method |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6274314B1 (en) * | 1998-04-02 | 2001-08-14 | Nyxis Neurotherapies, Inc. | Diagnostic assay for the modified nucleosides pseudouridine, 7-methyladenosine, or 1-methyladenosine |
| KR100565939B1 (en) * | 2003-11-28 | 2006-03-30 | 한국과학기술연구원 | Method for determination of nucleosides in urine with liquid chromatography-mass spectrometry |
-
2016
- 2016-01-20 CN CN201610038315.4A patent/CN105548407B/en active Active
Non-Patent Citations (4)
| Title |
|---|
| An improved method for glycosaminoglycan analysis by LC–MS/MS of urine samples collected on filter paper;Christiane Auray-Blais et al.;《Clinica Chimica Acta》;20121231;第413卷;第771-778页 * |
| Development of a filter paper method potentially applicable to mass and high-risk urinary screenings for Fabry disease;C. Auray-Blais · D. Cyr · K. Mills · R. Gigu`ere · R. Drouin;《JIMD Short Report》;20061231;第32卷;第1-6页 * |
| HPLC／ESI·Q-TOF MS鉴定淋巴癌患者尿液中的修饰核苷;李华雨等;《高等学校化学学报》;20090430;第30卷(第4期);第661-669页 * |
| Rapid Screening of High-Risk Patients for Disorders of Purine and Pyrimidine Metabolism Using HPLC-Electrospray Tandem Mass Spectrometry of Liquid Urine or Urine-soaked Filter Paper Strips;Tetsuya Ito et al.;《Clinical chemistry》;20001231;第46卷(第4期);摘要、表1、第446页左栏第1段至第448页左栏 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105548407A (en) | 2016-05-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105548407B (en) | The detection method of modified nucleoside in urine | |
| Stokes et al. | Development of a liquid chromatography–mass spectrometry method for the high-accuracy determination of creatinine in serum | |
| CN106093261B (en) | A kind of method that starch syrup is mixed in discriminating honey | |
| CN105548412A (en) | Method for measuring residual quantities of five aminoglycoside drugs in food simultaneously | |
| CN105675788B (en) | The method of progesterone and testosterone in high performance liquid chromatography tandem mass spectrum technology for detection saliva | |
| CN105181839A (en) | Method for detecting residual quantity of ivermectin in sheep muscle tissues by using liquid chromatograph/mass spectrometer with doramectin as internal standard substance | |
| CN104391068A (en) | Method for determining residual quantity of conventional bactericides in tobacco | |
| CN105510480A (en) | Detection method and application of 2-methylimidazole and 4-methylimidazole in baked food | |
| CN108362795A (en) | Content of homocysteine rapid detection method in dried blood spot | |
| CN105891364A (en) | Method and kit for detecting melatonin in saliva with HPLC-MS/MS (high performance liquid chromatography-tandem mass spectrometry) technology | |
| CN114720704B (en) | Kit and method for measuring free testosterone in serum | |
| CN102735764A (en) | Method for determining content of ribavirin in blood plasma | |
| CN110988193A (en) | Method for detecting advanced glycosylation end products in aquatic products | |
| CN103123345A (en) | Method for rapidly detecting phenoxyacetic acid herbicide in soil | |
| CN111505179A (en) | Method for detecting biopterin in marine water body | |
| CN102565252B (en) | Method for detecting content of homocysteine in blood or urine | |
| Rama et al. | Flow-through sensor with Fourier transform Raman detection for determination of sulfonamides | |
| CN103149289A (en) | Method for determining residual amount of 2, 4-D in tobacco | |
| CN109324139A (en) | Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf | |
| CN113466356A (en) | Sample pretreatment and detection method for determining pesticide residue content in cow milk | |
| CN109324140A (en) | Ribosylzeatin Solid Phase Extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf | |
| CN111366656A (en) | High performance liquid chromatography-tandem mass spectrometry method for detecting asomate residue in fruits and vegetables | |
| CN103512991A (en) | Automatic on-line analysis method for acrylamide mercapt uric acid metabolites in urine | |
| CN110763800A (en) | Method for detecting oxcarbazepine and 10, 11-dihydro-10-hydroxycarbazepine in blood | |
| CN109632986B (en) | Post-column derivatization detection method for sugar and sugar alcohol compounds |
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 |