CN104215685B - Enhanced reagent kit for qualitative and quantitative analysis detection and high-throughput screening of small molecules of pollutants and preparation method of enhanced reagent kit for qualitative and quantitative analysis detection and high-throughput screening of small molecules of pollutants - Google Patents
Enhanced reagent kit for qualitative and quantitative analysis detection and high-throughput screening of small molecules of pollutants and preparation method of enhanced reagent kit for qualitative and quantitative analysis detection and high-throughput screening of small molecules of pollutants Download PDFInfo
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- CN104215685B CN104215685B CN201410455796.XA CN201410455796A CN104215685B CN 104215685 B CN104215685 B CN 104215685B CN 201410455796 A CN201410455796 A CN 201410455796A CN 104215685 B CN104215685 B CN 104215685B
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Abstract
The invention relates to a preparation method of an enhanced reagent kit for the qualitative and quantitative analysis detection and the high-throughput screening of small molecules of pollutants. The enhanced reagent kit is used for detecting polarity pollutants in drinking water and during sewage treatment. The operation is easy and the efficiency is high. The preparation method of the enhanced reagent kit comprises the following steps: putting a sample (a single centrifugal pipe, a porous plate with 96 pores and a porous plate with 384 holes) under the room temperature for 10min, and adequately thawing the sample; adding a reagent QLW (comprising the following material compositions by weight percent: 50% to 100% of methanol and 1% to 5% of trifluoroacetic acid) with the identical volume at the temperature of minus 20 DEG C so as to obtain a mixture, acting for 15min, and acting for 5min in a water bath being 50 DEG C; centrifuging the mixture for more than 10min at the rotation speed of 3000rmp, and collecting liquid supernatant; adding a preparation reagent QLB1 with the same volume into the collected liquid supernatant; adequately mixing the preparation reagent QLB1, putting a solution, dispensing the solution on a target plate, standing for 10min and detecting the target plate, wherein the and volumetric ratio of the QLB1 comprises the following material compositions by weight percent: 50 to 70 percent of methanol, 30 to 50 percent of ionized water and 1 to 7 percent of trifluoroacetic acid. The reagent kit is simple in preparation method and relatively low in cost.
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 substrate
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, substrate energy absorption from laser
Biomolecule is passed to, and proton is obtained by proton translocation to biomolecule or from biomolecule in ionization process, and is made biology
The process of molecular ionization.Therefore it is a kind of Soft ionization techniques, it is adaptable to the measure of mixture and biomacromolecule.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 was directly proportional to the flight time of ion, detected ion.MALDI-TOF-MS have sensitivity it is high,
The features such as accuracy height and high resolution, a kind of strong analysis means of testing is provided for fields such as life sciences.
However, all matter parsed based on surface laser 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 is formed crystal.When
When irradiating crystal with laser, substrate energy absorption from laser, sample desorption, electric charge transfer occurs between substrate-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 reaching the flight of detector
Between it is different and be detected, i.e., be directly proportional to analyze ion by the ratio (M/Z) of the quality electric charge of ion to the flight time of ion,
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 parsed based on surface laser 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 analyses for being parsed based on surface laser in other words cannot
Put to good use and detect above in small molecule analysis. and molecular weight is most important to the mankind most to apply less than the small molecule of 1000da
One class chemical productss of value, such as more than 95% Medicine small molecule, all of aminoacid, 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 that often photometry sensitivity is relatively low, 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 detection and the high flux screening in small molecule.But need to consume substantial amounts of manpower and molten
Agent consumptive material, and a sample is from mass spectral analyses are prepared into again to the considerably long process of result 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 is required.
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
Prop up and the time.
Technical scheme：Pole in a kind of MALDI-TOF-MS detection drinking water and sewage disposal
The high flux screening test kit of contact scar thing small molecule, QLW material compositions and volume ratio are：50-100% methanol, 1-5% trifluoros
Acetic acid；QLB1 material compositions and volume ratio are：50-70% methanol, 30-50% ionized waters, 1-7% trifluoroacetic acids.
Polar contaminants in a kind of MALDI-TOF-MS detection drinking water and sewage disposal
The high flux screening test kit of small molecule, comprises the following steps that：Sample is placed into room temperature, is fully thawed, add -20 to spend peer
Long-pending reagent QLW, reacts 5 minutes in 50 degree of water-baths；After centrifugation, supernatant is collected；After taking supernatant, the system of equal volume is added
Standby reagent QLB1；After being sufficiently mixed, solution point is taken on target plate, sample detection after standing；QLW material compositions and volume ratio are：
50-100% methanol, 1-5% trifluoroacetic acids；QLB1 material compositions and volume ratio are：50-70% methanol, 30-50% ionized waters,
1-7% trifluoroacetic acids.
Preferred version is：QLW material compositions and volume ratio are：100% methanol, 1% trifluoroacetic acid；QLB1 material compositions and
Volume ratio is：50% methanol, 50% ionized water, 3% trifluoroacetic acid.
Beneficial effect：1st, preparation method is simple；2nd, cost is relatively low.
Specific embodiment
Apply the detection of the polar contaminants in drinking water and sewage disposal.It is simple to operate, efficiently.By sample (it is single from
Heart pipe, 96 hole porous plates, 384 hole porous plates) place room temperature 10 minutes, fully thaw.Add -20 reagents for spending equal volume
QLW (50-100% methanol, 1-5% trifluoroacetic acids), acts on 15 minutes, and acts on 5 minutes in 50 degree of water-baths.3000rmp from
After the heart 10 minutes, supernatant is collected.After taking supernatant, reagent preparation QLB1 of equal volume is added；After being sufficiently mixed, solution point is taken
On target plate, sample detection after standing 10 minutes；QLW material compositions and volume ratio are：50-100% methanol, 1-5% trifluoro second
Acid；QLB1 material compositions and volume ratio are：50-70% methanol, 30-50% ionized waters, 1-7% trifluoroacetic acids.
Claims (2)
1. during a kind of MALDI-TOF-MS detects drinking water and sewage disposal, polar contaminants are little
The high flux screening test kit of molecule, it is characterised in that by with reagent preparation QLB1 of QLW lysed sample and equal volume
Composition, QLW material compositions and volume ratio are：50-100% methanol, 1-5% trifluoroacetic acids；QLB1 material compositions and volume ratio are：
50-70% methanol, 30-50% ionized waters, 1-7% trifluoroacetic acids.
2. a kind of MALDI-TOF-MS detection drinking water described in claim 1 and sewage disposal
The high flux screening test kit of middle polar contaminants small molecule, it is characterised in that comprise the following steps that：Sample is placed into room temperature,
Fully thaw, add -20 reagent QLW for spending equal volume, react 5 minutes in 50 degree of water-baths；After centrifugation, supernatant is collected；
After taking supernatant, reagent preparation QLB1 of equal volume is added；After being sufficiently mixed, solution point is taken on target plate, after standing, sample introduction is examined
Survey；QLW material compositions and volume ratio are：50-100% methanol, 1-5% trifluoroacetic acids；QLB1 material compositions and volume ratio are：
50-70% methanol, 30-50% ionized waters, 1-7% trifluoroacetic acids.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410455796.XA CN104215685B (en) | 2014-09-09 | 2014-09-09 | Enhanced reagent kit for qualitative and quantitative analysis detection and high-throughput screening of small molecules of pollutants and preparation method of enhanced reagent kit for qualitative and quantitative analysis detection and high-throughput screening of small molecules of pollutants |
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| CN201410455796.XA CN104215685B (en) | 2014-09-09 | 2014-09-09 | Enhanced reagent kit for qualitative and quantitative analysis detection and high-throughput screening of small molecules of pollutants and preparation method of enhanced reagent kit for qualitative and quantitative analysis detection and high-throughput screening of small molecules of pollutants |
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| Publication Number | Publication Date |
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| CN104215685A CN104215685A (en) | 2014-12-17 |
| CN104215685B true CN104215685B (en) | 2017-04-19 |
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2014
- 2014-09-09 CN CN201410455796.XA patent/CN104215685B/en active Active
Non-Patent Citations (6)
| Title |
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| 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 * |
| Proton Sponge: A Novel and Versatile MALDI Matrix for the Analysis of Metabolites Using Mass Spectrometry;Rohit Shroff et.al;《Analytical Chemistry》;20091231;第81卷(第19期);第7954-7959页 * |
| 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;宋玉玲等;《中国科学：化学》;20111231;第41卷(第1期);第74-81页 * |
| 适用小分子化合物MALDI分析的基质研究;张森等;《化学进展》;20131225;第26卷(第1期);第158-166页 * |
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