CN110146452A - A method of mercury contamination of food is quickly detected based on ionic liquid - Google Patents
A method of mercury contamination of food is quickly detected based on ionic liquid Download PDFInfo
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Abstract
The present invention relates to a kind of methods of quickly detection Mercury In Food ion.This method includes three steps: (1) resolution of food samples to be measured；(2) mercury ion is extracted from food nitre solution liquid with ionic liquid；(3) ionic liquid is mixed with the deoxyribozyme reaction system with hydrogen peroxide enzymatic activity, detects the absorption value of ultraviolet-visible of the water phase at 418nm, the concentration of mercury ion is determined according to working curve.This method efficiently combines ionic liquid with highly selective ion enrichment ability and deoxyribozyme catalytic capability, effective fast enriching to mercury ion in the food digestion solution with superpower acidity and oxidisability is realized, the ionic liquid after mercury ion enrichment is directly used in the colorimetric detection of convenience, the mercury ion that sensitive and specific deoxyribozyme is catalyzed.This method quickly, it is easy, be readily applicable to the detection of mercury ion in the complex dielectrics such as soil, haze particle or be generalized to the detection of other heavy metal ion.
Description
Technical field
It is efficiently catalyzed with highly selective ion enrichment ability and deoxyribozyme the present invention relates to a kind of using ionic liquid
Ability, the method for quickly detecting Mercury In Food ion, belongs to technical field of analytical chemistry.
Background technique
Common heavy metal detection method has atomic emission spectrometry, atomic absorption spectrography (AAS), atomic fluorescence method, mass spectrum
Method etc..These are traditional high based on the requirement professional to operator of the detection method of large-scale instrument, and equipment is big and expensive, preceding
Processing is complicated, is unsuitable for field quick detection.The biosensor technology based on functional nucleic acid to grow up in recent years, tool
Have response quickly, it is easy to operate, be not necessarily to large scale equipment the advantages that, be concerned because it has a good application prospect.
Functional nucleic acid is that a kind of by what engineer or in-vitro screening obtained there is certain catalysis or specificity to know
The oligonucleotides of the short chain of other function.Functional nucleic acid mainly includes engineer's sequence, aptamer, ribozyme and deoxidation core
Several major class of enzyme.The most of biosensor for mercury ion detecting based on functional nucleic acid utilizes the richness of engineer
Oligonucleotide sequence containing thymidine.The specificity coordination of thymidine and mercury ion is that this kind of biology passes in oligonucleotides
The common detection mechanism of sensor (Journal of the American Chemical Society, 2006,128 (7): 2172-
2173), in conjunction with various signal transduction mechanism, realize to the fluorescence of mercury ion specificity (Analytical chemistry,
2012,84 (18): 8060-8066.), colorimetric (Analytical chemistry, 2008,80 (17): 6805-6808) and
Electrochemistry (Analytical chemistry, 2011,83 (17): 6896-6901) detection.Deoxidation is also used in minority report
Ribozyme is used for the detection of mercury ion.For example, the taking off with RNA shear active special to uranyl ion after Engineering Design
Oxygen ribozyme also reports fluorescence detection (Angewandte Chemie, 2007,119 (40): 7731- for mercury ion
7734.).The deoxyribozyme with catalase activity is subjected to Engineering Design for another example, if introducing in nucleic acid sequence
Dry thymine alkali bases.Mercury ion can be coordinated with these thymine alkali bases, to pass through the activity two for destroying deoxyribozyme
Level structure and make its inactivation, thereby realize the detection pattern (Analytical subtracted to the signal of mercury ion
chemistry,2009,81(6):2144-2149.).Although these methods all have very high sensitivity and selectivity,
Poor anti jamming capability is simply possible to use in the detection of mercury ion in these simple media of such as drinking water and ambient water, cannot achieve pair
The detection of mercury ion in this kind of complex dielectrics of food.Because the detection of Mercury In Food ion needs to carry out the resolution processing of sample,
The highly acid of nitre solution liquid and the testing conditions of oxidisability and sensor neutrality are incompatible, and other ingredients in digestion solution make
The appearance false positive of sensor or the response of false negative.
Ionic liquid is room temperature or is bordering on the substance being made of ion being in a liquid state at room temperature, also known as room temperature molten salt.From
Sub- liquid with the advantages that its good electric conductivity, wide electrochemical window, negligible vapour pressure and special dissolubility at
For the ideal substitute of conventional organic solvents.Ionic liquid is mainly characterized by: non-volatile, low melting point, wide liquid journey, strong
Electrostatic field, good electric conductivity, thermal conductivity and translucency, high refractive index, high heat capacity ratio, high thermal stability.Ionic liquid is answered
With extensive, wherein a big purposes is it to the unique selective extraction capacity of metal ion.Ionic liquid can be used for selective extraction
It takes Ag (I), Hg (II), Cd (II), Co (II), Ni (II), Fe (III), UO2 -, Co (II), Cd (II), Cu (II), Zn
A variety of common metal ions such as (II), Pb (II), Mn (II) (Separation Science and Technology,
2012,47(2):189-203；Journal of Separation Science,2018；41:373–384.).Mercury is as a kind of
Common heavy metal contaminants are had been reported that more using the research that ionic liquid carries out the selective extraction of mercury ion.Sieve in 2007
Gus seminar introduces thioether and thiourea functional group on hexafluorophosphoric acid alkyl imidazolium cation, enhance ionic liquid to mercury
The extracting power (Journal of Physical Chemistry C, 2007,111 (4): 1675-1682.) of ion.2008
Nian Puluo is that time project is combined into and has studied a series of ionic liquids to the extracting power of various metals, and discovery contains two sulphur
Two kinds of ionic liquids [MPYRROS2] of ehter bond and [MPIPS2] have very strong extracting power to mercury and copper ion, to other sun from
Sub- extraction efficiency very low (Industrial&Engineering Chemistry Research, 2007,47 (15): 5080-
5086.).The enrichment of ionic liquid is also used for the pre-treatment of the sample of the method for instrument detection.Ma Tingsi seminar in 2009
The mercury chelate in various water samples is extracted with butyl -3- methylimidazole hexafluorophosphate [BMIM] [PF6], later with cold original
Sub- absorption spectromtry concentration (Journal of Hazardous Materials, 2009,167:475-481).But it has no
The report of mercury ion in ionic liquid enriched food, any report associated with more not by it with sensor.
Summary of the invention
It is an object of the present invention to provide a kind of new methods that can quickly detect Mercury In Food ion.
A kind of method quickly detecting mercury contamination of food based on ionic liquid provided by the invention, including three steps:
Step 1: the resolution of food samples to be measured；Step 2: mercury ion is extracted from food nitre solution liquid with ionic liquid；Step
Rapid three, ionic liquid is mixed with the deoxyribozyme reaction system with hydrogen peroxide enzymatic activity, is qualitatively judged according to color
The concentration of mercury ion, or the absorption value of ultraviolet-visible of the detection water phase at 418nm, according to working curve determine mercury from
The concentration (content in food) of son.
Steps are as follows for specific experiment of the invention:
(1) drafting of working curve:
1) mercury ion of various concentration is added in ionic liquid, incubation with heat, ice bath is sudden cold, is centrifugated ionic liquid
Body；
2) buffer solution of certain density deoxyribozyme oligonucleotides, heating, slow cooling to room temperature are configured；Chlorination is added
Ferroheme is sufficiently mixed with deoxyribozyme oligonucleotides and is incubated for half an hour；
3) mixed solution of ionic liquid and mercury ion is added to the mixing of hemin and deoxyribozyme oligonucleotides
Solution is incubated for；
4) it is centrifuged, takes upper layer aqueous solution, catalysis reaction substrate and other reactive components is added；
5) with the UV absorption at ultraviolet specrophotometer measurement Product characteristics wavelength, working curve is drawn.
(2) selective measurement:
Solution containing mercury ion is replaced with to other metal ions of isoconcentration respectively, repeats the experiment of (1).
(3) ionic liquid is enriched with the extraction efficiency of various concentration mercury ion from buffer solution and 20% nitric acid respectively
Measurement:
The mercury ion of various concentration in 20% nitric acid is added in ionic liquid, incubation with heat, ice bath is sudden cold, and centrifugation is received
Collect supernatant.The concentration of mercury ion in supernatant is tested with atomic fluorescence spectrophotometer.Extraction efficiency (1- is calculated
(CInitially*VInitially-CAfter extraction*VAfter extraction)/CInitially*VInitially) * 100%.
(4) micro-wave digestion of sample: weighing sample and be placed in micro-wave diminishing pot, and digestion solution is added, and carries out micro-wave digestion,
Reagent blank is done simultaneously.Counteracting tank is uncapped after resolution end is cooling to be placed in graphite resolution instrument and is heated.It is with water that nitre solution liquid is dilute
It releases, for use.
(5) in sample mercury content measurement: a certain amount of ionic liquid is added in the digestion solution after above-mentioned dilution, is repeated
(1) other all operations in, finally measure ultraviolet absorption value of the sample at Product characteristics wavelength.According to (1) work obtained
The content of mercury ion in composition line computation sample.
This method efficiently mutually ties ionic liquid with highly selective ion enrichment ability with deoxyribozyme catalytic capability
It closes, realizes effective fast enriching to mercury ion in the food digestion solution with superpower acidity and oxidisability, mercury ion enrichment
Ionic liquid afterwards is directly used in the colorimetric detection of convenience, the mercury ion that sensitive and specific deoxyribozyme is catalyzed.Deoxidation core
Enzyme greatly improves the hydrogen peroxide enzymatic activity of hemin in conjunction with hemin.Quick catalysis hydrogen peroxide
It aoxidizes 2,2- connection (3- ethyl-benzothiazole -6- sulfonic acid) di-ammonium salts (ABTS) of nitrogen-two and generates the ABTS cation of green freely
Base (ABTS+·).The radical cation has a characteristic absorption peak at 418nm, and the catalytic activity of intensity and picodna is at just
Than.When not having mercury ion in food samples, deoxyribozyme in ion liquid abstraction deoxyribozyme reaction system makes deoxidation
Ribozyme inactivation；When there is mercury ion in food samples, the active site of ionic liquid is occupied by mercury ion, extracts deoxyribozyme
Reduced capability, deoxyribozyme activity keep, keep degree it is directly proportional to ion concentration of mercury in sample, thus realize pair
The quantitative detection of mercury ion.
This method has the advantage that 1) this method can be used in the quick detection of mercury contamination of food；2) biology is utilized
Sensor realizes the quantitative detection of mercury ion, does not need large-scale detecting instrument, easy to operate；3) it is carried out using ionic liquid
The selective enrichment of mercury ion in sample, selectivity is high, and ionic liquid requirement is few and can recycle；4) this method can be with
The detection of mercury ion suitable for soil and haze etc.；5) this method can be generalized to pair by using different ionic liquids
The detection of other metal ions；6) this method is easy to implement automatic operation.
Detailed description of the invention
Fig. 1 is the schematic diagram of the method for the present invention.
Fig. 2A and Fig. 2 B is the working curve diagram and gradient colorimetric photo of the method for the present invention, and wherein Fig. 2A is present invention side
The working curve of method, Fig. 2 B are gradient colorimetric photos, start to carry out within the 8th minute the measurement of absorbance value and take pictures in reaction.
Fig. 3 is the ion selectivity figure of the method for the present invention, wherein with Hg2+Difference with the absorbance of blank assay is hundred
/ hundred, the value added that absorbance after remaining metal ion is added is converted as percentage respectively.
Fig. 4 is N- octylpyridinium tetrafluoroborate to various concentration mercury ion in buffer solution and in 20% nitric acid
Extraction efficiency figure.
Fig. 5 is the test result figure that the method for the present invention uses different ionic liquid.
Fig. 6 is the test result figure that the method for the present invention uses different deoxyribozyme.
Specific embodiment
A kind of method quickly detecting mercury contamination of food based on ionic liquid of the invention, including three steps: step
One, the resolution of food samples to be measured；Step 2: mercury ion is extracted from food nitre solution liquid with ionic liquid；Step
Three, ionic liquid is mixed with the deoxyribozyme reaction system with hydrogen peroxide enzymatic activity, detects water phase at 418nm
Ultraviolet-visible absorption value, the concentration (content in food) of mercury ion is determined according to working curve.
As shown, Fig. 1 is the schematic diagram of the method for the present invention.Fig. 2 is the working curve diagram and gradient ratio of the method for the present invention
Color photo, wherein Fig. 2A is the working curve of the method for the present invention, and Fig. 2 B is gradient colorimetric photo, is started the 8th minute in reaction
It carries out the measurement of absorbance value and takes pictures.Fig. 3 is the ion selectivity figure of the method for the present invention, wherein with Hg2+With blank assay
The difference of absorbance is absolutely, respectively to convert the value added that absorbance after remaining metal ion is added for percentage.
Fig. 4 is extraction efficiency of the N- octylpyridinium tetrafluoroborate to various concentration mercury ion in buffer solution and in 20% nitric acid
Figure.Fig. 5 is the test result figure that the method for the present invention uses different ionic liquid.Fig. 6 is that the method for the present invention uses different deoxidation cores
The test result figure of enzyme.
As shown in Figure 1, when there is no mercury ion in food digestion solution sample, ion liquid abstraction deoxyribozyme reactant
Deoxyribozyme in system, inactivates deoxyribozyme, and reaction system does not develop the color；When having mercury ion in food digestion solution sample, from
The active site of sub- liquid is occupied by mercury ion, extracts the reduced capability of deoxyribozyme, and deoxyribozyme activity is kept, reactant
The aobvious green of system, the depth of color is directly proportional to ion concentration of mercury in sample, to realize the quantitative detection to mercury ion.
The drafting of 1. working curve of embodiment.
Mercury solution, the deoxyribozyme oligonucleotides EAD2 (5 '-arrived used in following all examples
CTGGGAGGGAGGGAGGGA-3 ') solution, hemin solution, 2,2- connection (the 3- ethyl-benzothiazole -6- sulphur of nitrogen-two
Acid) di-ammonium salts (ABTS) and hydrogenperoxide steam generator, with 4- hydroxyethyl piperazineethanesulfonic acid buffer solution A (25mM HEPES,
20mM KNO3,200 mM NaNO3,150mM NH4Cl, 0.025% (w/v) Triton X-100, and 1% (v/v)
It DMSO)) is retarder thinner.
As shown in Figure 1,10 μ L ionic liquids (N- octylpyridinium tetrafluoro boric acid) is taken to be added to 1mL various concentration mercury ion
In solution (0,0.5,1,2,5,7,10,50,100nM).10min, then the sudden cold 10min of ice bath are heated at 80 DEG C.8000r/min
The ionic liquid for being centrifuged bottom of the tube is taken out after being centrifuged 10min separation.Using buffer solution A as retarder thinner, 60 μ L, 0.5 μ is configured
M deoxyribozyme oligonucleotides EAD2 solution, 95 DEG C heated at constant temperature 6 minutes, then slow cooling is to 37 DEG C.As shown in Figure 1, above-mentioned
It is added in EAD2 solution hemin (15 μ L, 20 μM), 25 DEG C of incubations 90min, formation deoxyribozyme compound.2 μ L centrifugation
Resulting ionic liquid and 40 μ L deoxyribozyme compounds, 25 DEG C of incubation 30min.8000r/min centrifugation takes 35 μ L of upper layer de-
Oxygen ribozyme compound adds buffer solution A to be diluted to 100 μ L.Then 10 5 μ L of μ L ABTS and 15mM of 3.75mM is added simultaneously
Hydrogen peroxide.The UV absorption of 400-500nm wave band is scanned, with ultraviolet specrophotometer with the absorption at the 8th minute 418nm
Value draws working curve.
As a result as shown in Fig. 2, absorbance signal is positively correlated with ion concentration of mercury.It is linearly related preferably in low concentration section,
Color change when with the naked eye can recognize containing 0.5nM mercury ion.
The measurement of 2. selectivity of embodiment:
10 μ L N- octylpyridinium tetrafluoro boric acids are taken to be added to 1mL100nM different metal ions solution (Hg2+、Cr3+、Ni2 +、Ca2+、Pb2+、Cu2+、Fe3+、Fe2+、Cd2+、Zn2+、Mg2+) in.10min, the sudden cold 10min of ice bath are heated at 80 DEG C.8000r/
Min is centrifuged the N- octylpyridinium tetrafluoro boric acid that centrifugation bottom of the tube is taken out in 10min separation.Configure 60 μ L, 0.5 μM of deoxyribozyme widow
Nucleotide EAD2 solution, 95 DEG C heated at constant temperature 6 minutes, then slow cooling is to 37 DEG C.It is blood red that chlorination is added in above-mentioned EAD2 solution
Element (15 μ L, 20 μM), 25 DEG C of incubations 90min, formation deoxyribozyme compound.2 μ L are centrifuged resulting ionic liquid and 40 μ L are de-
Oxygen ribozyme compound, 25 DEG C of incubation 30min.8000r/min centrifugation, takes 35 μ L deoxyribozyme compound of upper layer, adds buffering molten
Liquid A is diluted to 100 μ L.Then the hydrogen peroxide of the 5 μ L of ABTS and 15mM of 10 μ L of 3.75mM is added simultaneously.Use ultraviolet spectrometry
Photometer tests the absorption value at the 8th minute 418nm.
As a result as shown in figure 3, this sensor has very strong specificity to mercury ion.Drawn in the presence of the mercury ion of 100nM
The increase of the absorbance value risen is 5-100 times of same concentrations other common heavy metals.Even in the presence of the mercury ion of 10nM
Signal intensity of the increase of caused absorbance value also than other common heavy metals of 100nM is big.
3. ionic liquid of embodiment from buffer solution and 20% nitric acid in be enriched with various concentration mercury ion extraction efficiency
Measurement:
It takes 10 μ L N- octylpyridinium tetrafluoro boric acids to be added to 1mL to contain in the water of various concentration mercury ion or 20% nitre
Acid solution (0,0.5,1,2,5,7,10,50,100nM).10min, the sudden cold 10min of ice bath are heated at 80 DEG C.8000r/min from
The N- octylpyridinium tetrafluoro boric acid of centrifugation bottom of the tube is taken out in heart 10min separation.Supernatant is collected, with atomic fluorescence spectrophotometric
The concentration of mercury ion in measurement examination supernatant.Instrument condition 270V, 25mA.15s is measured, 0.5s is postponed.Current-carrying is 5%HCl,
Reducing agent is 2%NaBH4, extraction efficiency (1- (C is calculated in 0.5%NaOHInitially*VInitially-CAfter extraction*VAfter extraction)/CInitially*VInitially)*
100%.
As a result as shown in figure 4, N- octylpyridinium tetrafluoro boric acid no matter to the mercury in buffer solution or in 20% nitric acid from
Son obtains good effect of extracting.Can reach 50% or more extraction yield in the low concentration range, and with mercury from
Sub- concentration increases extraction efficiency and gradually increases.The equal Gao Qiang of extraction efficiency in all tests, in the buffer solution of subacidity
Acid condition, the especially extraction to rather high concentration mercury ion.In the buffer solution of subacidity, the mercury ion of high concentration
The extraction efficiency of (50nM, 100nM) reaches 94% or more；Under strong acidic environment, extraction efficiency is significantly reduced, extraction efficiency
About 75%.In the buffer solution of subacidity and under strong acidic condition, to the extraction efficiency of the mercury ion (1-10nM) of low concentration
Relatively, in the range of 50%-73%.
The testing result of mercury ion content in the food samples of 4. the method according to the invention of embodiment measurement
It weighs about 0.5g flesh of fish sample to be placed in polytetrafluoroethylene (PTFE) (PTFE) micro-wave diminishing pot, 5mL concentrated nitric acid, 2mL is added
Hydrogen peroxide, by 120 DEG C of 1min, 160 DEG C of 10min, 180 DEG C of 10min carry out micro-wave digestion, while doing reagent blank.Resolution knot
Counteracting tank is uncapped after beam is cooling and is placed in 120 DEG C of heating 60min in graphite resolution instrument, removes part spent acid and nitrogen oxides.
With 5 times of volume of water dilute samples.In sample nitre solution liquid after taking 10 μ L N- octylpyridinium tetrafluoro boric acids to be added to 1mL dilution.
10min, then the sudden cold 10min of ice bath are heated at 80 DEG C.8000 r/min centrifugation 10min separation takes the N- octyl pyrrole of centrifugation bottom of the tube
Pyridine tetrafluoro boric acid.The ultrapure water of 5 times of volumes is added, is beaten with rifle, ultrasonic 5min, sudden cold 10min centrifugation removes supernatant, weight
Again twice.Configure 60 μ L, 0.5 μM of deoxyribozyme oligonucleotides EAD2 solution, 95 DEG C heated at constant temperature 6 minutes, slow cooling is to 37 DEG C.
It is added in above-mentioned EAD2 solution hemin (15 μ L, 20 μM), 25 DEG C of incubations 90min, formation deoxyribozyme compound.2
μ L is centrifuged resulting ionic liquid and 40 μ L deoxyribozyme compounds, 25 DEG C of incubation 30min.8000r/min centrifugation, takes upper layer
35 μ L deoxyribozyme compounds, add buffer solution A to be diluted to 100 μ L.Then simultaneously be added 10 μ L of 3.75mM ABTS and
The hydrogen peroxide of 5 μ L of 15mM.The absorption value at the 8th minute 418nm is tested with ultraviolet specrophotometer, it is bent further according to work
Line determines the concentration (content in food) of mercury ion.
As a result show such as table 1, this sensor can preferably suitable for various food samples various concentration range mercury ion
The testing result deviation of quickly detection, sensor detection results and national standard method is generally more slightly higher than actual concentration within 10%
A bit.But entire effect deviation is little, can satisfy the needs of field screening substantially.
Table 1: the method according to the invention measurement food samples in mercury ion content testing result and with country mark
The comparison of quasi- method testing result
Error is obtained by 3 repetition experimental calculations.
5. the method for the present invention of embodiment uses the test result of different ionic liquid:
All experiment reagents and method employed in the present embodiment are identical as described in embodiment 1, only by ionic liquid
Body is changed to 1- octyl -3- methylimidazole hexafluorophosphate and pyrrolidines dithiocarbonic acid by N- octylpyridinium tetrafluoro boric acid respectively
Ammonium (APDC) or 1- butyl -3- methylimidazole hexafluorophosphate and APDC.
As shown in figure 5, deoxyribozyme catalytic activity with higher, the product of generation exists when being free of ionic liquid
Absorption at 418nm is relatively strong (uppermost ultraviolet-visible absorption spectroscopy line).When addition is free of the above two ion of mercury ion
When any one of liquid, the catalytic activity of deoxyribozyme significantly reduces (nethermost two ultraviolet-visible absorption spectroscopies
Line).When any one of the above two ionic liquid containing mercury ion is added, the catalytic activity of deoxyribozyme is between most
High between minimum catalytic activity (intermediate two ultraviolet-visible absorption spectroscopy lines).As a result illustrate that method of the invention has
Universality also can achieve similar invention effect using other ionic liquids (including the combination with ligand), may be implemented pair
The detection of mercury ion.
6. the method for the present invention of embodiment uses the test result of different deoxyribozyme:
All experiment reagents and method employed in the present embodiment are identical as described in embodiment 1, only by deoxidation core
Enzyme oligonucleotides EAD2 (5 '-CTGGGAGGGAGGGAGGGA-3 ') is changed to AT11A (5 '-respectively
AGGGACGGGAT11AGGGACGGGA-3').As shown in fig. 6, deoxyribozyme is with higher to urge when being free of ionic liquid
Change activity, absorption of the product of generation at 418nm is relatively strong (No. 3, color is bottle green).It is upper without mercury ion when being added
When stating any one of ionic liquid, the catalytic activity of deoxyribozyme significantly reduces (No. 1, colourless).Contain mercury ion when being added
Ionic liquid when, the catalytic activity of deoxyribozyme (No. 2, green) between highest and minimum catalytic activity.As a result it says
Bright method of the invention has universality, also can achieve similar invention effect using other deoxyribozyme, may be implemented pair
The detection of mercury ion.When AT11A is changed to EADT11 (5 '-CTGGGAGGGTTTTTTTTTTTAGGGAGGGA-3 '), phenomenon
It is identical.
Claims (5)
1. a kind of method for quickly detecting mercury contamination of food based on ionic liquid, including three steps: Step 1: food to be measured
The resolution of sample；Step 2: mercury ion is extracted from food nitre solution liquid with ionic liquid；Step 3: ionic liquid with
Deoxyribozyme reaction system mixing with hydrogen peroxide enzymatic activity, the concentration of mercury ion is qualitatively judged according to color, or
Person detects the absorption value of ultraviolet-visible of the water phase at 418nm, and the concentration of mercury ion is determined according to working curve.
2. the method for detection mercury contamination of food according to claim 1, which is characterized in that step 1 is the microwave of sample
Resolution: weighing sample and be placed in micro-wave diminishing pot, and digestion solution is added, and carries out micro-wave digestion, while doing reagent blank, resolution terminates
Counteracting tank is uncapped to be placed in graphite resolution instrument after cooling and is heated, is diluted nitre solution liquid with water, for use.
3. the method for detection mercury contamination of food according to claim 1 or 2, which is characterized in that in step 2, take 10 μ L
N- octylpyridinium tetrafluoro boric acid is added in the sample nitre solution liquid after 1mL dilution, heats 10min at 80 DEG C, then ice bath is sudden cold
10min, 8000r/min centrifugation 10min separation take the N- octylpyridinium tetrafluoro boric acid of centrifugation bottom of the tube, and the super of 5 times of volumes is added
Pure water is beaten with rifle, ultrasonic 5min, and sudden cold 10min centrifugation removes supernatant, is repeated twice.
4. the method for detection mercury contamination of food according to claim 1 or 2, which is characterized in that in step 3, configuration 60
0.5 μM of deoxyribozyme oligonucleotides EAD2 solution of μ L, 95 DEG C heated at constant temperature 6 minutes, slow cooling is to 37 DEG C, in above-mentioned EAD2 solution
Middle addition hemin, 25 DEG C of incubation 90min form deoxyribozyme compound, and 2 μ L are centrifuged resulting ionic liquid and 40 μ L
Deoxyribozyme compound, 25 DEG C of incubation 30min, 8000r/min centrifugations, takes 35 μ L deoxyribozyme compound of upper layer, adds buffering molten
Liquid A is diluted to 100 μ L, and the hydrogen peroxide of the 5 μ L of ABTS and 15mM of 10 μ L of 3.75mM is then added simultaneously.
5. the method for detection mercury contamination of food according to claim 1 or 2, which is characterized in that the working curve
Drafting include the following steps: 1) in ionic liquid be added various concentration mercury ion, incubation with heat, ice bath is sudden cold, centrifugation
Separating ionic liquid；
2) buffer solution of certain density deoxyribozyme oligonucleotides, heating, slow cooling to room temperature are configured；It is blood red that chlorination is added
Element is sufficiently mixed with deoxyribozyme oligonucleotides and is incubated for half an hour；
3) mixed solution of ionic liquid and mercury ion is added to the mixed solution of hemin and deoxyribozyme oligonucleotides,
It is incubated for；
4) it is centrifuged, takes upper layer aqueous solution, catalysis reaction substrate and other reactive components is added；
5) with the UV absorption at ultraviolet specrophotometer measurement Product characteristics wavelength, working curve is drawn.
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| CN201810138308.0A CN110146452B (en) | 2018-02-10 | 2018-02-10 | Method for rapidly detecting mercury content in food based on ionic liquid |
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| CN110106176A (en) * | 2019-05-14 | 2019-08-09 | 贵州理工学院 | A kind of tetra- serobilas of the G--hemin DNA enzymatic and its measuring method of colorimetric method for determining silver ion and mercury ion |
| CN110514609A (en) * | 2019-09-26 | 2019-11-29 | 武汉海关技术中心 | The analyzing detecting method of lead metal element in solid food sample |
| CN110567775A (en) * | 2019-09-26 | 2019-12-13 | 武汉海关技术中心 | Solid food sample pretreatment method based on ionic liquid |
| CN112710652A (en) * | 2019-10-24 | 2021-04-27 | 中国科学院理化技术研究所 | Ultra-sensitive mercury pollution colorimetric detection method based on copper sulfide nano material |
| CN113267604A (en) * | 2021-05-17 | 2021-08-17 | 山东省海洋化工科学研究院 | Method for detecting metallic mercury in food |
| CN113788788A (en) * | 2021-07-23 | 2021-12-14 | 浙江工业大学 | Fluorescent ionic liquid and synthesis method and application thereof |
| CN114199977A (en) * | 2021-11-23 | 2022-03-18 | 江苏大学 | Electrochemical heavy metal rapid detection method based on electric drive pre-enrichment |
| CN114456438A (en) * | 2022-01-24 | 2022-05-10 | 四川大学 | Composite material for enriching viruses in droplets and application thereof |
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| CN110106176A (en) * | 2019-05-14 | 2019-08-09 | 贵州理工学院 | A kind of tetra- serobilas of the G--hemin DNA enzymatic and its measuring method of colorimetric method for determining silver ion and mercury ion |
| CN110514609A (en) * | 2019-09-26 | 2019-11-29 | 武汉海关技术中心 | The analyzing detecting method of lead metal element in solid food sample |
| CN110567775A (en) * | 2019-09-26 | 2019-12-13 | 武汉海关技术中心 | Solid food sample pretreatment method based on ionic liquid |
| CN110514609B (en) * | 2019-09-26 | 2022-08-02 | 武汉海关技术中心 | Analysis and detection method for lead metal element in solid food sample |
| CN110567775B (en) * | 2019-09-26 | 2022-08-12 | 武汉海关技术中心 | Solid food sample pretreatment method based on ionic liquid |
| CN112710652A (en) * | 2019-10-24 | 2021-04-27 | 中国科学院理化技术研究所 | Ultra-sensitive mercury pollution colorimetric detection method based on copper sulfide nano material |
| CN113267604A (en) * | 2021-05-17 | 2021-08-17 | 山东省海洋化工科学研究院 | Method for detecting metallic mercury in food |
| CN113788788A (en) * | 2021-07-23 | 2021-12-14 | 浙江工业大学 | Fluorescent ionic liquid and synthesis method and application thereof |
| CN113788788B (en) * | 2021-07-23 | 2023-12-05 | 浙江工业大学 | Fluorescent ionic liquid and synthesis method and application thereof |
| CN114199977A (en) * | 2021-11-23 | 2022-03-18 | 江苏大学 | Electrochemical heavy metal rapid detection method based on electric drive pre-enrichment |
| CN114199977B (en) * | 2021-11-23 | 2023-08-22 | 江苏大学 | Electrochemical heavy metal rapid detection method based on electric drive pre-enrichment |
| CN114456438A (en) * | 2022-01-24 | 2022-05-10 | 四川大学 | Composite material for enriching viruses in droplets and application thereof |
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