CN104215682B - One kind is applied to the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit and its preparation - Google Patents
One kind is applied to the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit and its preparation Download PDFInfo
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- CN104215682B CN104215682B CN201410452868.5A CN201410452868A CN104215682B CN 104215682 B CN104215682 B CN 104215682B CN 201410452868 A CN201410452868 A CN 201410452868A CN 104215682 B CN104215682 B CN 104215682B
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Abstract
It is a kind of to be applied to the detection of amino acid molecular qualitative and quantitative analysis and the preparation of high flux screening agent box, comprise the following steps that：Any lysate carries out cracking and prepares sample, after taking supernatant, adds the reagent preparation QLA of equal volume；It is sufficiently mixed；Solution point is taken on target plate, sample detection after being stood under ventilation；QLA material compositions and volume ratio are：30% ionized water, 30 50% methyl alcohol, 20 40% normal propyl alcohols, 1 5% formic acid；This reagent box preparation method is simple, cost is relatively low.
Description
Technical field
This technology belongs to analytical chemistry applications field, and in particular to urface laser and resolves field of mass spectrometry.
Background technology
Novel soft ionization biological mass spectrometry is a kind of new surface laser resolves mass spectrum developed in recent years, wherein with matrix
Assisted Laser Desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) is most important example, and the present invention obtained Nobel
Chemistry prize.Mainly it is made up of two parts：MALDI (MALDI) and TOF
(TOF).The principle of MALDI is with laser irradiating sample and substrate formed cocrystallization film, matrix energy absorption from laser
Biomolecule is passed to, proton is obtained by proton translocation to biomolecule or from biomolecule in ionization process, and make biology
The process of molecular ionization.Therefore it is a kind of Soft ionization techniques, it is adaptable to the measure of mixture and large biological molecule.The original of TOF
Reason is that ion accelerates to fly over dirft tube under electric field action, is detected and surveys according to the flight time difference for reaching detector
The mass-to-charge ratio (M/Z) for determining ion is directly proportional to the flight time of ion, detects ion.MALDI-TOF-MS have sensitivity it is high,
The degree of accuracy is high and the features such as high resolution, for the fields such as life science provide a kind of strong analysis means of testing.
However, all matter based on surface laser parsing headed by MALDI-TOF-MS
Spectrum is required for by sample preparation on special target plate, and its operation is that in substrate molecule and sample dispersion formed into crystal.When
When irradiating crystal with laser, matrix energy absorption from laser, sample desorption, electric charge transfer occurs between matrix-sample to be made
Ionized sample molecule is obtained, the sample of ionization flies over the tof tube of vacuum under electric field action, during according to the flight for reaching detector
Between it is different and be detected, i.e., be directly proportional to analyze ion to the flight time of ion by the ratio between the quality electric charge of ion (M/Z),
And measure the molecular weight of sample molecule.Focal issue is that the molecular weight of general substrate molecule, in 1000-3000da, is caused
Excessive background mass spectra peak.This just determines that all such mass spectral analyses based on surface laser parsing may be only available for big point
Son detection, such as protein, long-chain polypeptide, nucleic acid, macromolecular material etc., and may not apply to small molecule detection (molecular weight
Less than 1000da), unique advantage such as high sensitivity/high flux of mass spectral analysis based on surface laser parsing cannot in other words
Put in small molecule analysis detection that the small molecule of and molecular weight less than 1000da is most important to the mankind most to apply above to good use
One class chemical products of value, such as more than 95% Medicine small molecule, all of amino acid, vitamin etc. and human lives
Healthy related chemical substance.
Currently available technology is detected for small molecule and screening also rests on spectroscopic methodology detection, for example fluorescence radiation detection or
Person derives can be with the substrate of fluoroscopic examination, but such detection requires that target small molecule molecular structure has activity group, separately
One drawback is exactly relatively low often photometry sensitivity, can't detect micro little molecular concentration.Liquid chromatography mass spectrometric (LC-MS) and gas phase
The having begun to of mass spectrum (GC-MS) applies the detection in small molecule and high flux screening.But need to consume substantial amounts of manpower and molten
Agent consumptive material, and a sample is from mass spectral analysis is prepared into again to the result considerably long process of needs is gone out, it is different as requested,
One sample of analysis needs 30-60 minutes from sample preparation to result is taken, and far can not meet modern biotechnology medicine development
High flux screening requirement.
The content of the invention
Goal of the invention：Find that a kind of preparation method is simple, lower-cost nhancing ionization buffer liquid preparation method, saving is opened
Branch and time.
Technical scheme：One kind is applied to the detection of amino acid molecular qualitative and quantitative analysis and high flux screening kit, by inciting somebody to action
The reagent preparation QLA of lysed sample and equal volume is constituted, and QLA material compositions and volume ratio are：Ionized water:Methyl alcohol:Positive third
Alcohol:Formic acid=30:30-50:20-40:1-5.
It is a kind of to be applied to the detection of amino acid molecular qualitative and quantitative analysis and the preparation of high flux screening kit, specific steps
It is as follows：Any lysate carries out cracking and prepares sample, after taking supernatant, adds the reagent preparation QLA of equal volume；It is sufficiently mixed；
Solution point is taken on target plate, sample detection after being stood under ventilation；QLA material compositions and volume ratio are：Ionized water:Methyl alcohol:Just
Propyl alcohol:Formic acid=30:30-50:20-40:1-5.
Preferred scheme is：QLA material compositions and volume ratio are：Ionized water:Methyl alcohol:Normal propyl alcohol:Formic acid=30:30:20:1.
Beneficial effect：1st, preparation method is simple；2nd, cost is relatively low.
Brief description of the drawings
Fig. 1 utilize the invention detects that amino acid molecular mass spectrogram
Specific embodiment
This technology method is adapted to amino acid molecular, because with multiple amino on amino acid molecular structure, naturally with reference to target
Plate, adds peptide molecule of the invention once dense ionization buffer solution so that most amino acid moleculars fly close to 100% ionization and gasification
To mass detector.
Sample is prepared by certain cracking mode first, after taking supernatant, the reagent preparation QLP (ions of equal volume is added
Water:Methyl alcohol:Normal propyl alcohol:Formic acid=30: 30-50: 20-40: 1-5).After being sufficiently mixed, 0.5 microlitre of solution point is taken on target plate,
Sample introduction after standing 10 minutes under ventilation.
The result of detection is as shown in Figure 1.
Claims (2)
1. the high flux screening that a kind of MALDI-TOF-MS for being applied to amino acid molecular is detected
Kit, it is characterised in that constituted by by the reagent preparation QLA of lysed sample and equal volume, QLA material compositions and volume
Than for：30% ionized water, 30-50% methyl alcohol, 20-40% normal propyl alcohol 1-5% formic acid.
2. the high flux screening that a kind of MALDI-TOF-MS for being applied to amino acid molecular is detected
The preparation of kit, it is characterised in that comprise the following steps that：Any lysate carries out cracking and prepares sample, after taking supernatant, adds
The reagent preparation QLA of equal volume；It is sufficiently mixed；Solution point is taken on target plate, sample detection after being stood under ventilation；QLA things
Matter is constituted and volume ratio is：30% ionized water, 30-50% methyl alcohol, 20-40% normal propyl alcohols, 1-5% 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 CN104215682A (en) | 2014-12-17 |
| CN104215682B true CN104215682B (en) | 2017-06-16 |
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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)
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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 |
|---|
| Acid– base-driven matrix-assisted mass spectrometry for targeted metabolomics;Rohit Shroff et.al;《PNAS》;20090623;第106卷(第25期);第10092-10096页 * |
| N-(1-Naphthyl) Ethylenediamine Dinitrate: A New Matrix for Negative Ion MALDI-TOF MS Analysis of Small Molecules;Rui Chen et.al;《American Society for Mass Spectrometry》;20120619;第23卷;第1454-1460页 * |
| Peptidomics of CNS-Associated Neurohemal Systems of Adult Drosophila melanogaster: A Mass Spectrometric Survey of Peptides from Individual Flies;REINHARD PREDEL et.al;《THE JOURNAL OF COMPARATIVE NEUROLOGY》;20041231;第474卷;第379-392页 * |
| 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》.2011,第41卷(第1期),第74-81页. * |
| 张森等.适用小分子化合物MALDI分析的基质研究.《化学进展》.2013,第26卷(第1期),第158-166页. * |
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| CN104215682A (en) | 2014-12-17 |
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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. |