CN104215682A - Kit applied to amino acid molecule qualitative and quantitative analysis and detection and high throughput screening and preparation of kit - Google Patents
Kit applied to amino acid molecule qualitative and quantitative analysis and detection and high throughput screening and preparation of kit Download PDFInfo
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- CN104215682A CN104215682A CN201410452868.5A CN201410452868A CN104215682A CN 104215682 A CN104215682 A CN 104215682A CN 201410452868 A CN201410452868 A CN 201410452868A CN 104215682 A CN104215682 A CN 104215682A
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Abstract
The invention discloses a method for preparing a kit applied to amino acid molecule qualitative and quantitative analysis and detection and high throughput screening. The method comprises the following specific steps: performing lysis to prepare a sample by using any lysis solution, taking the supernatant, and adding a preparation agent QLA of the same volume; fully mixing; adding the solution onto a target plate, standing at a ventilating part, and detecting, wherein the QLA comprises the following substances in percentage by volume: 30 percent of ionized water, 30-50 percent of methanol, 20-40 percent of normal propyl alcohol and 1-5 percent of formic acid. The kit is simple in preparation method and low in cost.
Description
Technical field
This technology belongs to analytical chemistry applications field, is specifically related to surface laser and resolves field of mass spectrometry.
Background technology
It is a kind of novel soft ionization biological mass spectrometry that development in recent years is got up that surface laser resolves mass spectrum, wherein with Matrix Assisted Laser Desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) for most important example, the present invention obtained Nobel chemistry Prize.Form primarily of two parts: substance assistant laser desorpted ionized ion gun (MALDI) and time of flight mass analyzer (TOF).The principle of MALDI irradiates sample and substrate formed cocrystallization film with laser, matrix absorbs energy transferring to biomolecule from laser, in ionization process, proton translocation is obtained proton to biomolecule or from biomolecule, and the process that biomolecule is ionized.Therefore it is a kind of Soft ionization techniques, is applicable to the mensuration of potpourri and biomacromolecule.The principle of TOF is that ion accelerates to fly over dirft tube under electric field action, and according to the flight time arriving detecting device is different, the detected mass-to-charge ratio (M/Z) namely measuring ion was directly proportional to the flight time of ion, detects ion.MALDI-TOF-MS have highly sensitive, accuracy is high and resolution high, for the fields such as life science provide a kind of strong analytical test means.
But all mass spectrums of resolving based on surface laser headed by Matrix Assisted Laser Desorption ionization time of flight mass spectrometry all need by sample preparation on special target plate, its operation is that sample dispersion is formed crystal in substrate molecule.When irradiating crystal with laser, matrix absorbs energy from laser, sample desorption, there is Charger transfer between matrix-sample and make ionized sample molecule, the sample of ionization flies over the tof tube of vacuum under electric field action, different and be detected according to the flight time arriving detecting device, to be namely directly proportional to the flight time of ion by the ratio (M/Z) of the quality electric charge of ion and to analyze ion, and record the molecular weight of sample molecule.Focal issue is just that the molecular weight of general substrate molecule is at 1000-3000da, causes excessive background mass spectra peak.This just determines all mass spectrophotometry that this type of is resolved based on surface laser can only be applicable to large Molecular Detection, such as protein, long-chain polypeptide, nucleic acid, macromolecular material etc., and Small molecular detection (molecular weight is less than 1000da) can not be applied to, the advantage that the high sensitivity/high flux etc. of the mass spectrophotometry of resolving based on surface laser is in other words unique cannot be put to good use on small molecule analysis detects. and the Small molecular that molecular weight is less than 1000da is the class chemical products to the most important most using value of the mankind, the Medicine small molecule of such as more than 95%, all amino acid, vitamin etc. relevant chemical substance healthy with human lives.
Currently available technology detects for Small molecular and screening also rests on spectroscopic methodology detection, such as fluorescence radiation detects or derives can the substrate of fluoroscopic examination, but this type of testing requirement target Small molecular molecular structure has photolytic activity group, another drawback is exactly that often photometry sensitivity is lower, can't detect the Small molecular concentration of trace.The starting of liquid chromatography mass spectrometric (LC-MS) and gaseous mass spectrum (GC-MS) is applied in micromolecular detection and high flux screening.But need to consume a large amount of manpowers and solvent consumptive material, and a sample needs quite to grow a process to going out result from being prepared into mass spectrophotometry again, different as requested, analyze a sample and need 30-60 minute from sample preparation to taking result, the high flux screening requirement of modern biotechnology medicine development can not be met far away.
Summary of the invention
Goal of the invention: find a kind of preparation method simple, lower-cost enhancing ionization buffer liquid and preparation method thereof, reduce expenses and the time.
Technical scheme: one is applied to amino acid molecular qualitative and quantitative analysis and detects and high flux screening kit, form by by the reagent QLA for preparing of lysed sample and equal volume, QLA material composition and volume ratio are: 30% ionized water, 30-50% methyl alcohol, 20-40% n-propanol 1-5% formic acid.
Be applied to a preparation for the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit, concrete steps are as follows: arbitrary lysate carries out cracking for sample, and after getting supernatant, what add equal volume prepares reagent QLA; Abundant mixing; Get solution point on target plate, sample detection after leaving standstill under ventilation; QLA material composition and volume ratio are: 30% ionized water, 30-50% methyl alcohol, 20-40% n-propanol, 1-5% formic acid.
Preferred version is: QLA material composition and volume ratio are: 30% ionized water, 30% methyl alcohol, 20% n-propanol 1% formic acid.
Beneficial effect: 1, preparation method is simple; 2, cost is lower.
Accompanying drawing explanation
The amino acid molecular mass spectrogram that Fig. 1 utilizes the present invention to detect
Embodiment
This technical method is applicable to amino acid molecular, because with multiple amino on amino acid molecular structure, natural in target plate, adds peptide molecule of the present invention once dense ionization damping fluid, makes most amino acid molecular fly to mass detector close to 100% ionization and gasification.
First prepare sample by certain cracking mode, after getting supernatant, what add equal volume prepares reagent QLP (70-90% ionized water, 10-30% methyl alcohol, 1-5% formic acid).After abundant mixing, get 0.5 Al of Solution point on target plate, under ventilation, leave standstill sample introduction after 10 minutes.
The result detected is as shown in Figure 1.
Claims (3)
1. one kind is applied to the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit, it is characterized in that, form by by the reagent QLA for preparing of lysed sample and equal volume, QLA material composition and volume ratio are: 30% ionized water, 30-50% methyl alcohol, 20-40% n-propanol 1-5% formic acid.
2. be applied to a preparation for the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit, it is characterized in that, concrete steps are as follows: arbitrary lysate carries out cracking for sample, and after getting supernatant, what add equal volume prepares reagent QLA; Abundant mixing; Get solution point on target plate, sample detection after leaving standstill under ventilation; QLA material composition and volume ratio are: 30% ionized water, 30-50% methyl alcohol, 20-40% n-propanol, 1-5% formic acid.
3. be according to claim 1ly applied to a kind of amino acid molecular qualitative and quantitative analysis that is applied to and detect and the preparation of high flux screening kit, it is characterized in that, QLA material composition and volume ratio are: 30% ionized water, 30% methyl alcohol, 20% n-propanol 1% formic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410452868.5A CN104215682B (en) | 2014-09-09 | 2014-09-09 | One kind is applied to the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit and its preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410452868.5A CN104215682B (en) | 2014-09-09 | 2014-09-09 | One kind is applied to the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit and its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104215682A true CN104215682A (en) | 2014-12-17 |
| CN104215682B CN104215682B (en) | 2017-06-16 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201410452868.5A Active CN104215682B (en) | 2014-09-09 | 2014-09-09 | One kind is applied to the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit and its preparation |
Country Status (1)
| Country | Link |
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| CN (1) | CN104215682B (en) |
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2014
- 2014-09-09 CN CN201410452868.5A patent/CN104215682B/en active Active
Non-Patent Citations (5)
| Title |
|---|
| REINHARD PREDEL ET.AL: "Peptidomics of CNS-Associated Neurohemal Systems of Adult Drosophila melanogaster: A Mass Spectrometric Survey of Peptides from Individual Flies", 《THE JOURNAL OF COMPARATIVE NEUROLOGY》 * |
| ROHIT SHROFF ET.AL: "Acid– base-driven matrix-assisted mass spectrometry for targeted metabolomics", 《PNAS》 * |
| RUI CHEN ET.AL: "N-(1-Naphthyl) Ethylenediamine Dinitrate: A New Matrix for Negative Ion MALDI-TOF MS Analysis of Small Molecules", 《AMERICAN SOCIETY FOR MASS SPECTROMETRY》 * |
| 宋玉玲等: "基质辅助激光解析电离-飞行时间质谱分析寡核苷酸G-四链体", 《中国科学：化学》 * |
| 张森等: "适用小分子化合物MALDI分析的基质研究", 《化学进展》 * |
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| CN104215682B (en) | 2017-06-16 |
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Effective date of registration: 20211122 Address after: 210000 east of floor 5, building 3, intelligent manufacturing Industrial Park (Zhicheng Park), No. 6, Zhida Road, Jiangbei new area, Nanjing, Jiangsu Province Patentee after: Nanjing Pinsheng Medical Technology Co.,Ltd. Patentee after: Fuzhou Pinsheng Medical Technology Co., Ltd Address before: 430040 Room 303, building 1, phase II, Strait venture City, Qixiong Road, Dongxihu District, Wuhan City, Hubei Province Patentee before: WUHAN QUALITY LIFE TECHNOLOGY Co.,Ltd. |