CN106568865B - Urea [13C] abundance detection method - Google Patents
Urea [13C] abundance detection method Download PDFInfo
- Publication number
- CN106568865B CN106568865B CN201610984856.6A CN201610984856A CN106568865B CN 106568865 B CN106568865 B CN 106568865B CN 201610984856 A CN201610984856 A CN 201610984856A CN 106568865 B CN106568865 B CN 106568865B
- Authority
- CN
- China
- Prior art keywords
- urea
- acetonitrile
- abundance
- take
- test solution
- 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
Abstract
The invention discloses a kind of urea [13C] abundance detection method, comprising the following steps: S1. take urea [13C] particle, finely ground, precision weighs 0.5g, sets in 20ml measuring bottle, adds acetonitrile appropriate, and it is 9-15 minutes ultrasonic, it lets cool to room temperature, adds dilution in acetonitrile to scale, shake up, take solution, with 10000r/min centrifugation 9-15 minutes, take supernatant as test solution.S2. 1 μ l of test solution is taken to inject liquid chromatograph-mass spectrometer, record the chromatogram and quasi-molecular ion (12C16ON2H5+, 13C16ON2H5+, 13C18ON2H5+ generated, mass number is respectively 61.0402,62.0435,64.0478, be respectively designated as A, B, C) ion signal response intensity, calculate to get；Urea [13C] abundance calculation formula are as follows: X=(B+C/A+B+C) × 100%, present invention employs exclusive chromatographic condition and Mass Spectrometry Conditions to reduce the human error factor in detection process so that detection is more easy, the reproducibility of testing result is more preferable, and testing result is more accurate.
Description
Technical field
The present invention relates to medical sciences, more particularly to a kind of method of accurate detection urea [13C] abundance.
Background technique
Urea [13C] breath test is used to check the infection of helicobacter pylori, and principle is will to pass through stable nuclide
The substrate of [13C] label introduces body (major way is oral), utilizes the final generation of isotope-ratio mass spectrometer detection substrate
The variation for thanking to product 13CO2 comes metabolic response and physiology course in research aircraft body, to requirement >=99% of 13C urea abundance, because
This need to detect the 13C urea abundance of urea [13C] test agents, existing liquid-quality detection method, the reproducibility of testing result
Bad, testing result error is big.
Summary of the invention
The present invention is to solve the above problem of existing detection technique, will test condition optimizing, this detection method includes following
Step:
S1. urea [13C] particle is taken, finely ground, precision weighs 0.5g, sets in 20ml measuring bottle, adds acetonitrile appropriate, ultrasonic 9-
It 15 minutes, lets cool to room temperature, adds dilution in acetonitrile to scale, shake up, take solution, with 10000r/min centrifugation 9-15 minutes, take
Supernatant is as test solution.
S2. take 1 μ l of test solution inject liquid chromatograph-mass spectrometer, record generation chromatogram and quasi-molecule from
Son (12C16ON2H5+, 13C16ON2H5+, 13C18ON2H5+, mass number are respectively 61.0402,62.0435,64.0478, point
Be not denoted as A, B, C) ion signal response intensity, calculate to get；The calculation formula of 13C abundance are as follows: X=(B+C)/(A+B+
C) ×100%。
Preferably, selecting chromatographic condition: using HILIC chromatographic column (2.1*100mm, 1.7 μm), mobile phase A is acetonitrile, stream
The ammonium acetate solution that dynamic phase B is 1mmol/L, gradient elution (0-4min, 95%A → 50%A;4-4.5min,50%A→95%A;
4.5-8min, 95%A), flow velocity 0.2-0.4ml/min, 33-36 DEG C of column temperature；Mass Spectrometry Conditions: electron spray positively ionized (ESI
+) detection, scanning range m/z60~121, capillary voltage 3.0KV, orifice potential 8V, 96-101 DEG C of source temperature, desolventizing gas
440-455 DEG C of temperature, Desolvention gas velocity 740-805L/h.
This detection method is more easier than existing isotope mass spectrometer detection method, reduces artificial in detection process
Error component；The reproducibility of testing result is more preferable, and testing result is more accurate.
Specific embodiment
Embodiment one
Detection method includes the following steps for urea [13C] abundance:
S1. urea [13C] particle is taken, finely ground, precision weighs 0.5g, sets in 20ml measuring bottle, adds acetonitrile appropriate, ultrasonic 9-
It 15 minutes, lets cool to room temperature, adds dilution in acetonitrile to scale, shake up, take solution, with 10000r/min centrifugation 9-15 minutes, take
Supernatant is as test solution.
S2. it takes 1 μ l of test solution to inject liquid chromatogram-level four bars flight time mass spectrum combined instrument, records the color of generation
Spectrogram and quasi-molecular ion (12C16ON2H5+, 13C16ON2H5+, 13C18ON2H5+, mass number is respectively 61.0402,
62.0435,64.0478, A, B, C are respectively designated as) ion signal response intensity, calculate to get urea [13C] abundance
Calculation formula are as follows: X=(B+C)/(A+ B+C) × 100%.
More specifically, it selects chromatographic condition: using HILIC chromatographic column (2.1*100mm, 1.7 μm), mobile phase A is second
Nitrile, Mobile phase B are the ammonium acetate solution of 1mmol/L, gradient elution (0-4min, 95%A → 50%A;4-4.5min,50%A→95%
A;4.5-8min, 95%A), flow velocity 0.2-0.4ml/min, 33-36 DEG C of column temperature；Mass Spectrometry Conditions: electron spray positively ionized
(ESI+) it detects, scanning range m/z60~121, capillary voltage 3.0KV, orifice potential 8V, 96-101 DEG C of source temperature, precipitation
440-455 DEG C of degree of agent temperature, Desolvention gas velocity 740-805L/h.
The above is only the ranges that preferred embodiments of the present invention, the present invention that therefore, it cannot be limited according to are implemented, i.e., according to the present invention
Equivalence changes made by description and decoration, all should belong to the present invention covering in the range of.
Claims (1)
1. a kind of urea [13C] abundance detection method, it is characterised in that: the following steps are included:
S1. urea [13C] particle is taken, finely ground, precision weighs 0.5g, sets in 20ml measuring bottle, and add acetonitrile appropriate, ultrasonic 9-15 points
Clock is let cool to room temperature, is added dilution in acetonitrile to scale, is shaken up, take solution, with 10000r/min centrifugation 9-15 minutes, take supernatant
As test solution；
S2. it takes 1 μ l of test solution to inject liquid chromatograph-mass spectrometer, records the chromatogram and quasi-molecular ion of generation
12C16ON2H5+, 13C16ON2H5+, 13C18ON2H5+, mass number are respectively 61.0402,62.0435,64.0478, respectively
Be denoted as the ion signal response intensity of A, B, C, calculate to get；The calculation formula of urea [13C] abundance are as follows: X=(B+C/A+
B +C) ×100%；
Further include selecting chromatographic condition: using HILIC chromatographic column 2.1*100mm, 1.7 μm, mobile phase A is acetonitrile, and Mobile phase B is
The ammonium acetate solution of 1mmol/L, gradient elution 0-4min, 95%A → 50%A;4-4.5min,50%A→95%A;4.5-8min,
95%A, flow velocity 0.2-0.4ml/min, 33-36 DEG C of column temperature；Mass Spectrometry Conditions: electron spray positively ionized ESI+ detection scans model
Enclose m/z60~121, capillary voltage 3.0KV, orifice potential 8V, 96-101 DEG C of source temperature, desolvation temperature 440-455
DEG C, Desolvention gas velocity 740- 805L/h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610984856.6A CN106568865B (en) | 2016-11-09 | 2016-11-09 | Urea [13C] abundance detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610984856.6A CN106568865B (en) | 2016-11-09 | 2016-11-09 | Urea [13C] abundance detection method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106568865A CN106568865A (en) | 2017-04-19 |
| CN106568865B true CN106568865B (en) | 2019-07-09 |
Family
ID=58540634
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610984856.6A Active CN106568865B (en) | 2016-11-09 | 2016-11-09 | Urea [13C] abundance detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106568865B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101811992A (en) * | 2010-04-02 | 2010-08-25 | 上海化工研究院 | Preparation method for semicarbazide hydrochloride labeled by stable isotope 13C and 15N |
| CN102053127A (en) * | 2010-11-11 | 2011-05-11 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Method for analyzing small molecules in biological sample quantitatively by adopting LC-MS (Liquid Chromatogram-Mass Spectrum) technology |
| CN103913525A (en) * | 2014-04-01 | 2014-07-09 | 山东省分析测试中心 | Method for detecting 17 plasticizers in white spirit by high performance liquid chromatography-tandem mass spectrometry |
| CN104122339A (en) * | 2013-04-25 | 2014-10-29 | 上海化工研究院 | Isotopic abundance detection method for D, 13C or 15N labeled organic compounds |
| CN105372340A (en) * | 2014-08-29 | 2016-03-02 | 重庆华邦制药有限公司 | Method of determining low-content paricalcitol through high performance liquid chromatography-tandem mass spectrometry method and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SE0400197D0 (en) * | 2004-01-29 | 2004-01-29 | Amersham Biosciences Ab | Method and system for reducing total sample complexity |
-
2016
- 2016-11-09 CN CN201610984856.6A patent/CN106568865B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101811992A (en) * | 2010-04-02 | 2010-08-25 | 上海化工研究院 | Preparation method for semicarbazide hydrochloride labeled by stable isotope 13C and 15N |
| CN102053127A (en) * | 2010-11-11 | 2011-05-11 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Method for analyzing small molecules in biological sample quantitatively by adopting LC-MS (Liquid Chromatogram-Mass Spectrum) technology |
| CN104122339A (en) * | 2013-04-25 | 2014-10-29 | 上海化工研究院 | Isotopic abundance detection method for D, 13C or 15N labeled organic compounds |
| CN103913525A (en) * | 2014-04-01 | 2014-07-09 | 山东省分析测试中心 | Method for detecting 17 plasticizers in white spirit by high performance liquid chromatography-tandem mass spectrometry |
| CN105372340A (en) * | 2014-08-29 | 2016-03-02 | 重庆华邦制药有限公司 | Method of determining low-content paricalcitol through high performance liquid chromatography-tandem mass spectrometry method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106568865A (en) | 2017-04-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103563043B (en) | System and corresponding method thereof for the chemico-analytic particularly medical domain with the calibration to instrument response of sample amounts | |
| CN105223264B (en) | A kind of simulation internal standard method, apparatus of mass spectrum quantitative analysis and application | |
| Zhang et al. | Integration of data-dependent acquisition (DDA) and data-independent high-definition MSE (HDMSE) for the comprehensive profiling and characterization of multicomponents from Panax japonicus by UHPLC/IM-QTOF-MS | |
| de la Mora | Ionization of vapor molecules by an electrospray cloud | |
| CN106290681A (en) | The method for quick of Multi-pesticide residues and pre-treating method thereof in tomato sauce | |
| Berns et al. | The CAPTAIN detector and physics program | |
| CN107526096B (en) | A method of amendment temperature is to SiPM gain effects | |
| Ryde et al. | Gamma-ray burst spectral correlations: photospheric and injection effects | |
| Ko et al. | Abundance variation at the vicinity of an active region and the coronal origin of the slow solar wind | |
| CN106568865B (en) | Urea [13C] abundance detection method | |
| Kakkad et al. | BASS XXXI: Outflow scaling relations in low redshift X-ray AGN host galaxies with MUSE | |
| CN106990174A (en) | SVOC quick determination methods in a kind of cloth velvet sample based on full volatilization head space | |
| CN109470764A (en) | Isotope ratio mass spectral analysis | |
| CN108387655A (en) | A method of for detecting persticide residue in Radix Ophiopogonis | |
| Chen et al. | Process development for the decoloration of Panax notoginseng extracts: a design space approach | |
| BESIII collaboration | Observation of a Neutral Structure near the D (D) over-bar* Mass Threshold in e (+) e (-)->(D (D) over-bar*)(0) pi (0) at root s= 4.226 and 4.257 GeV | |
| Winkel et al. | The Close AGN Reference Survey (CARS)-Tracing the circumnuclear star formation in the super-Eddington NLS1 Mrk 1044 | |
| Xiang et al. | Simultaneous detection of eight active components in Radix Tinosporae by ultra high performance liquid chromatography coupled with electrospray tandem mass spectrometry | |
| Sherma | High-performance liquid chromatography/mass spectrometry analysis of botanical medicines and dietary supplements: a review | |
| Erdogdu et al. | Simulation of Cluster Dynamics of Proton-Bound Water Clusters in a High Kinetic Energy Ion-Mobility Spectrometer | |
| Nikola et al. | Star formation in m51 triggered by galaxy interaction | |
| Fausch et al. | Monitoring space weather with a sensitive 1 u cubesat mass spectrometer | |
| Scheffler et al. | PNJL model analysis of the Roberge-Weiss transition endpoint at imaginary chemical potential | |
| CN107543746A (en) | The pre-treating method and quantitative detecting method of the endogenous plant hormone of three class difference chemical property in a kind of plant sample | |
| CN107589181A (en) | A kind of detection method of antelope's horn |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20191111 Address after: 225300 No. 801 Health Avenue, Taizhou City, Jiangsu Province, 36 (Pharmaceutical City) Patentee after: JIANGSU HUAGEN TAILAI BIOTECHNOLOGY CO., LTD. Address before: 102200, room 2102, block B, No. 9, front road, Changping District science and Technology Park, Beijing Co-patentee before: JIANGSU HUAGEN TAILAI BIOTECHNOLOGY CO., LTD. Patentee before: Boran Pharmaceutical Co., Ltd., Beijing |