CN106404734B - The method that silver nanoclusters detect potassium concentration infering diing time in vitreous humor - Google Patents
The method that silver nanoclusters detect potassium concentration infering diing time in vitreous humor Download PDFInfo
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Abstract
The invention belongs to medical jurisprudence technical fields, and in particular to a kind of method that silver nanoclusters detect potassium concentration infering diing time in vitreous humor.The method of the present invention is to synthesize the DNA-AgNCs, system specific recognition K with photoluminescent property using the Novel DNA chain containing potassium ion aptamers as template+Make silver nanoclusters fluorescent quenching, standard curve is drawn according to the variation of fluorescence, vitreous humor to be measured is added in DNA-AgNCs solution, potassium ion in vitreous humor causes silver nanoclusters fluorescence to change, compared with standard curve, potassium concentration in tested vitreous humor is obtained, by K in vitreum sample+Concentration and known death time draw out standard curve, as a result have good uniformity with the death time of report and vitreous humor potassium ion relationship.The present invention can be used for legal medical expert's corrupt corpse, have high degree of specificity and sensitivity, easy to operate, quick, detection that is low in cost and can be applied to potassium content in other body fluid and water environment.
Description
Technical field
The invention belongs to medical jurisprudence technical fields, and in particular to potassium concentration in a kind of silver nanoclusters detection vitreous humor
And then the method for infering diing time.
Background technique
Potassium ion (K+) it is one of the most abundant cation of people's in-vivo content, there is important physiological function to organism.
For the normal acid-base balance and osmotic pressure for keeping body, participates in various protein and glycometabolism institute is required；It is to maintain cell newly old
Metabolism adjusts body fluid pervious pressure, maintains one of acid-base balance and the important electrolyte of holding cellular stress function.Its content is machine
The very important index of body physiological activity, while the contents level of potassium ion can be used for Diagnosing Cardiac, kidney etc. in blood, urine
The disease of aspect and the prognosis for judging disease.
In addition, in forensic identification, since malpractice etc. happens occasionally；Especially electrolyte disturbance, cardiac muscle cell's function
Have with the concentration of cations some inside and outside cell membrane than relatively straightforward relationship, wherein most with the influence of calcium ion and potassium concentration
Obviously.Wherein especially with K in vitreous humor+Content is medical jurisprudence research hotspot；Human body after death, K in vitreous humor+Content be in
Regularity rises, extremely close with death time relationship, passes through K in measurement vitreous humor+Content come when inferring human body death
Between；K in vitreous humor is had proven to simultaneously+Content is by outside contamination, the putrefaction of dead body and such environmental effects very little.Therefore, quasi-
Really measurement K+Content has extremely important meaning to deduction Postmortem Interval.
K is measured in the prior art+The method of content mainly has: chemical assay, isotopic mass spectrometry, flame spectrometry,
Atom spectrophotometry, enzyme process and ion selective electrode method etc..Wherein common detection methods are ion selective electrode method and fire
Flame photometry.
(1) flame spectrometry: this method returned using excited atom in flame when being down to ground state the spectral intensity that emits into
Row content analysis is a kind of emission spectrometry method, can detect vitreous humor, urine, serum, the K in cerebrospinal fluid+Content.Often
The quantitative approach seen includes internal standard method and external standard method, and wherein external standard method general operation error is larger, and is of little use；Internal standard method will be marked
The internal standard that same concentrations are added in quasi- liquid and sample is measured, and is usually added into lithium internal standard, then measures lithium/potassium current ratio
Value, rather than the electric current of independent measurement potassium ion, can reduce error in this way, improve the accuracy of measurement.Its shortcoming is that fire
In the detection process, sensitivity is influenced flame photometry by gaseous-pressure, the pressure of combustion-supporting gas, sample sample introduction speed, and sample is added
It to be mixed well after distilled water, the sample and actual value otherwise analyzed have certain deviation.In addition, the maximum deficiency of flame spectrometry exists
In it is used be the combustion gas such as propane, bring security risk to laboratory.
(2) chemical assay: ring crown compound mainly is answered using ionophore and is measured, due to having size inside ring
Different cavity structure, internal oxygen atom electronics is to can be with metal ion-chelant.Different size of cavity alternative combines
The metal ion of different-diameter, to measure ion concentration.But anion, solvent, crown compound concentration, thickness of liquid film and
The factors such as temperature are affected to migration rate of the metal ion in compound, are easy to cause measurement result inaccurate.
(3) urine, blood, K+ determination in water ion selective electrode method: are carried out on special instrument.Its principle are as follows:
Ion selective electrode is a kind of chemical sensor, in a certain range, fixed ionic activity given in current potential and solution
Logarithm is in linear relation, measures liquor sample using the linear relationship, the color and turbidity of solution generally will not be to measurements
As a result have an impact.This method is divided into directly measurement and Indirect Determination again, measures at present frequently with the latter.Due to Indirect Determination
It is measured after determinand is diluted, gained ionic activity is closer to ion concentration.The electrode mainly used has: solid phase electrode, liquid
Membrane electrode, glass-membrane electrode etc..But these electrodes can be with using the time and aging, price are more expensive.
Summary of the invention
In view of the problems of the existing technology, the present invention provides potassium concentration in a kind of silver nanoclusters detection vitreous humor
The method of infering diing time, it is therefore an objective to potassium concentration is measured by the DNA-AgNCs fluorescence probe synthesized based on DNA chain,
So that K+ determination it is easier, more rapidly, it is more acurrate.
The method of potassium concentration infering diing time in silver nanoclusters provided by the invention detection vitreous humor, according to
Lower step carries out:
(1) it prepares fluorescent DNA-AgNCs: designing containing potassium ion aptamers and can load the DNA chain of AgNCs first,
AgNO is added in DNA chain3NaHB is added after oscillation in solution4Solution is put into refrigerator 3 hours after vibrating again, obtains brown color liquid,
That is fluorescent DNA-AgNCs；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared: preparing various concentration potassium salt soln, is added into step
Suddenly in the fluorescent DNA-AgNCs solution of (1), its fluorescence intensity at 630nm is measured after reacting a period of time, obtains potassium ion
The standard curve of concentration and fluorescence intensity；
(3) actual glass body fluid sample potassium concentration detect: by the vitreous humor to be measured of known death time be added to
In the fluorescent DNA-AgNCs solution of step (1), its fluorescence intensity at 630nm is measured after reacting a period of time, according to preparation
The potassium concentration and Standardization curve for fluorescence intensity of step (2) obtain K in institute's test sample sheet+Concentration；
(4) potassium concentration and death time standard curve are prepared: according to the known death time measured in step (3)
Vitreous humor K+The death time of concentration and known sample, drafting linear relationship chart, obtained potassium concentration and death time
Standard curve；
(5) corrupt corpse: the vitreous humor K to be measured of unknown death time is measured+Concentration, according to the potassium of step (4)
Ion concentration and death time standard curve, to be inferred to the death time of institute's test sample sheet.
Wherein, described containing potassium ion aptamers and the DNA chain of AgNCs be loaded to be capable of forming the G- tetramer
DNA chain is selected from nucleotide sequence table id NO.1, nucleotide sequence table id NO.2, nucleotide sequence table id NO.3, nucleotide
Sequence table ID NO.4, nucleotide sequence table id NO.5, nucleotide sequence table id NO.6, nucleotide sequence table id NO.7, core
Nucleotide sequence table id NO.8, nucleotide sequence table id NO.9, nucleotide sequence table id NO.10, nucleotide sequence table id
NO.11, nucleotide sequence table id NO.12, DNA sequence dna shown in nucleotide sequence table id NO.13.
The DNA chain and AgNO3、NaHB4Molar concentration rate is 1:2~10:2~10, preferably 1:6:6.
The pH value of the fluorescence AgNCs solution is 6-7.5.
The sylvite includes potassium chloride, potassium nitrate and potassium bromide and other potassium-containing compounds.
The solution to be measured is vitreous humor, blood, urine or other liquid of human or animal.
The reaction time of the step (2) and step (3) is 0.1~4 hour.
It is described to be used to the accurate supposition death time in forensic identification using potassium concentration in silver nanoclusters detection liquid.
The method is for detecting other compositions in vitreous humor, including ATP, hypoxanthine, magnesium ion.
Compared with prior art, the features of the present invention and beneficial effect are:
The basic principle of the method for potassium concentration infering diing time in silver nanoclusters detection vitreous humor of the invention
It is that potassium concentration is measured based on DNA chain synthetic DNA-AgNCs fluorescence probe, i.e., is formed using the N atom in DNA monomer with Ag
Stronger coordinate bond issues hyperfluorescence after connecting DNA-AgNCs by amino specificity, is by AgNO in the present invention3、NaHB4It presses
Certain proportion is coupled on target dna by chemical bond, and fluorescent DNA-AgNCs is obtained after Physical temperature-lowering, the subsequent fluorescence nano
Race specific chelating K again+Make its fluorescent quenching, K can be measured according to the variation of fluorescence+Concentration.Potassium ion solution to be measured is dripped
In above-mentioned fluorescent DNA-AgNCs solution, detect its change in fluorescence, by compared with obtained standard curve, obtain by
Survey the Potassium Level of body fluid.
The invention has the advantages that
(1) present invention using DNA chain specific recognition and combines K+, the G- tetramer conformation of regulation is formed, it can not be by it
The influence of his ion has high specificity to potassium ion.
(2) present invention uses DNA-AgNCs as fluorescence probe, to K+The G- tetramer conformation of regulation is very sensitive, tool
There is higher sensitivity.
(3) the DNA-AgNCs fluorescence probe based on DNA chain that the present invention designs, only needs Fluorescence Spectrometer, and cost is relatively low,
It can be widely popularized.
(4) of the invention to be used for K+The condition that the DNA-AgNCs fluorescence probe preparation process based on DNA chain of detection requires
Easy to control, operation is succinct, low energy consumption, is process for cleanly preparing, can be produced in batches.
(5) of the invention to be used for K+The DNA-AgNCs fluorescence probe based on DNA chain of detection, agents useful for same is common
Chemical reagent it is safe and non-toxic, it is environment friendly and pollution-free, can be widely applied.
(6) agents useful for same type of the present invention is less, simple operations, quick and low in cost；Material stability is good, when can be long
Between store, it is ensured that measurement effect.
(7) K+ determination method provided by the present invention can not only measure human body, blood, urine, glass in animal body
K in the body fluid such as glass body fluid+Concentration can also be measured the potassium concentration in other samples such as water quality, be widely used.
(8) it is provided by the present invention measurement vitreous humor in potassium concentration method, can quickly and accurately measure potassium from
Sub- concentration, applies to forensic identification for fluorescent technique for the first time, accurately infers the dead death time provider for legal medical expert worker
Method.
Detailed description of the invention
Fig. 1 is fluorescence intensity schematic diagram of the measurement containing potassium solution at 630nm when the present invention prepares standard curve；
Fig. 2 is the canonical plotting obtained according to Fig. 1；
Fig. 3 is the canonical plotting in the embodiment of the present invention 1；
Fig. 4 is the canonical plotting in the embodiment of the present invention 2；
Fig. 5 is the canonical plotting in the embodiment of the present invention 3.
Specific embodiment
Below by way of specific implementation case, the present invention will be described, but scope of protection of the present invention is not limited thereto.
Embodiment 1:
The method of potassium concentration infering diing time is according to following in the silver nanoclusters detection vitreous humor of the present embodiment
Step carries out:
(1) it prepares fluorescent DNA-AgNCs: the DNA solution of 50 μM of 90 μ L being taken, with 12000rpm, to take 50 μ L under the conditions of 4 DEG C
The AgNO of 500 μM of 54 μ L is added to centrifuge tube in bottom liquid in centrifuge tube3Solution and 102 μ L secondary waters, shake at room temperature
It swings 30 minutes；Then, 54 μ L, 500 μM of NaHB prepared with ice water are wherein being added4Solution, concussion are put into 4 DEG C of ice after five minutes
Case stores 3 hours, obtains brown color liquid, i.e. fluorescent DNA-AgNCs solution, and the pH of solution is 7.4, is sub-packed in centrifuge tube, often
30 μ LDNA-AgNCs solution are added dropwise in part, spare；
DNA chain nucleotides sequence used is classified as DNA sequence dna shown in nucleotide sequence table id NO.6 in this example；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared: compound concentration 0.1,0.4,0.8,1.2,1.8,
3.0, the potassium nitrate solution of 5.0,7.5,10.0,20.0mM is added into the fluorescent DNA-AgNCs solution of step (1), to
After solution reaction one minute, its fluorescence intensity at 630nm is measured, draws solution K+The linear pass of concentration and fluorescence intensity
Standard curve is made in system；
(3) actual glass body fluid sample potassium concentration detected: by 20 μ L vitreous humors to be measured (known dead 13.2 hours
Obtaining afterwards) sample solution is added into the fluorescent DNA-AgNCs solution of step (1), and the dilution of 100 μ L secondary waters is added, and 1 minute
Its fluorescence intensity at 630nm is measured afterwards.
(4) potassium concentration and death time standard curve are prepared: according to the known death time measured in step (3)
Vitreous humor K+The death time of concentration and known sample measures the vitreous humor K of different death times a series of+Concentration is drawn
The standard curve of potassium concentration and death time is made in linear relationship chart；
(5) K in institute's test sample sheet corrupt corpse: is obtained according to the standard curve of preparation step (2)+Concentration is 8.6mM；
The potassium concentration and death time standard curve of step (4), to infer that the death time for obtaining detected sample is 13.6
Hour, it is consistent substantially with the actual sample death time situation of the present embodiment.The vitreous humor sample solution standard is taken again
EDTA titration is measured, and obtains K+Concentration is 8.71mM；Measuring result error is less than 2%.Measurement result has good
Consistency, illustrate the method for the present invention can quantitative measurment various concentration K+, concentration ranges are 0.1~20.0mM.
Embodiment 2:
The method of potassium concentration infering diing time is according to following in the silver nanoclusters detection vitreous humor of the present embodiment
Step carries out:
(1) it prepares fluorescent DNA-AgNCs: the DNA solution of 50 μM of 100 μ L being taken, with 12000rpm, to take 60 μ under the conditions of 4 DEG C
The AgNO of 500 μM of 108 μ L is added to centrifuge tube in L bottom liquid in centrifuge tube3Solution and 204 μ L secondary waters, at room temperature
Concussion 30 minutes；Then, 108 μ L, 500 μM of NaHB prepared with ice water are wherein being added4Solution, concussion are put into 4 DEG C after five minutes
Refrigerator storage 3 hours, brown color liquid, i.e. fluorescent DNA-AgNCs solution are obtained, the pH of solution is 7.4, it is sub-packed in centrifuge tube,
Every part of 50 μ L DNA-AgNCs solution of dropwise addition, it is spare；
DNA chain nucleotides sequence used is classified as DNA sequence dna shown in nucleotide sequence table id NO.6 in this example；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared: compound concentration 0.1,0.8,2.4,5.0,10.0,
18.0,20 μ L potassium nitrate solutions are added to the fluorescent DNA-of step (1) potassium nitrate solution of 25.0,30.0,40.0mM
In AgNCs solution and the dilution of 100 μ L secondary waters is added, after solution reaction one minute, it is strong to measure its fluorescence at 630nm
Degree draws solution K+Standard curve is made in the linear relationship of concentration and fluorescence intensity；
(3) actual glass body fluid sample potassium concentration detected: by 20 μ L vitreous humors to be measured (known dead 19.5 hours
Obtaining afterwards) sample is added into the fluorescent DNA-AgNCs solution of step (1), and the dilution of 100 μ L secondary waters is added, it is surveyed after 1 minute
Measure its fluorescence intensity at 630nm.
(4) potassium concentration and death time standard curve are prepared: according to the known death time measured in step (3)
Vitreous humor K+The death time of concentration and known sample measures the vitreous humor K of different death times a series of+Concentration is drawn
The standard curve of potassium concentration and death time is made in linear relationship chart；
(5) K in institute's test sample sheet corrupt corpse: is obtained according to the standard curve of preparation step (2)+Concentration is
11.8mM；The potassium concentration and death time standard curve of step (4) obtain the death time of detected sample to infer
It is 18.9 hours, is consistent with the actual sample death time situation of the present embodiment.The vitreous humor sample standard is taken again
EDTA titration is measured, and obtains K+Concentration is 11.67mM；Measuring result error is less than 1%.Measurement result has good
Consistency, illustrate the method for the present invention can quantitative measurment various concentration K+, concentration ranges are 0.1~40.0mM.
Embodiment 3:
The method of potassium concentration infering diing time is according to following in the silver nanoclusters detection vitreous humor of the present embodiment
Step carries out:
(1) it prepares fluorescent DNA-AgNCs: the DNA solution of 50 μM of 100 μ L being taken, with 12000rpm, to take 60 μ under the conditions of 4 DEG C
The AgNO of 500 μM of 108 μ L is added to centrifuge tube in L bottom liquid in centrifuge tube3Solution and 204 μ L secondary waters, at room temperature
Concussion 30 minutes；Then, 108 μ L, 500 μM of NaHB prepared with ice water are wherein being added4Solution, concussion are put into 4 DEG C after five minutes
Refrigerator storage 3 hours, brown color liquid, i.e. fluorescent DNA-AgNCs solution are obtained, the pH of solution is 7.4, it is sub-packed in centrifuge tube,
Every part of 50 μ L DNA-AgNCs solution of dropwise addition, it is spare；
DNA chain nucleotides sequence used is classified as DNA sequence dna shown in nucleotide sequence table id NO.6 in this example；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared: compound concentration 0.1,0.8,2.4,5.0,10.0,
18.0,20 μ L potassium nitrate solutions are added to the fluorescent DNA-of step (1) potassium nitrate solution of 25.0,30.0,40.0mM
In AgNCs solution and the dilution of 100 μ L secondary waters is added, after solution reaction one minute, it is strong to measure its fluorescence at 630nm
Degree draws solution K+Standard curve is made in the linear relationship of concentration and fluorescence intensity；
(3) actual glass body fluid sample potassium concentration detects: by 20 μ L vitreous humors to be measured (after known dead 6 hours
Obtaining) sample is added into the fluorescent DNA-AgNCs solution of step (1), and the dilution of 100 μ L secondary waters is added, it is measured after 1 minute
Its fluorescence intensity at 630nm.
(4) potassium concentration and death time standard curve are prepared: according to the known death time measured in step (3)
Vitreous humor K+The death time of concentration and known sample measures the vitreous humor K of different death times a series of+Concentration is drawn
The standard curve of potassium concentration and death time is made in linear relationship chart；
(5) K in institute's test sample sheet corrupt corpse: is obtained according to the standard curve of preparation step (2)+Concentration is 4.2mM；
The potassium concentration and death time standard curve of step (4), to infer that the death time for obtaining detected sample is 6.3 small
When, it is consistent with the actual sample death time situation of the present embodiment.The EDTA titration of the vitreous humor sample standard is taken again
It is measured, obtains K+Concentration is 4.1mM；Measuring result error is less than 1%.Measurement result has good consistency, explanation
The method of the present invention can quantitative measurment various concentration K+, concentration ranges are 0.1~40.0mM.
Embodiment 4:
The detection method of the present embodiment is same as Example 1, only changes the DNA chain in embodiment into nucleotides sequence and is classified as core
DNA sequence dna shown in nucleotide sequence table id NO.1, final measurement equally may be implemented to K+The quantitative determination of concentration, measurement
The vitreous humor K of difference death times a series of+Concentration, drafting linear relationship chart, obtained potassium concentration and death time
Standard curve measures the vitreous humor K to be measured of unknown death time+Concentration, it is bent according to potassium concentration and death time standard
Line, to be inferred to the death time of institute's test sample sheet.
Embodiment 5:
The detection method of the present embodiment is same as Example 2, only changes the DNA chain in embodiment into nucleotides sequence and is classified as core
DNA sequence dna shown in nucleotide sequence table id NO.2, final measurement equally may be implemented to K+The quantitative determination of concentration, measurement
The vitreous humor K of difference death times a series of+Concentration, drafting linear relationship chart, obtained potassium concentration and death time
Standard curve measures the vitreous humor K to be measured of unknown death time+Concentration, it is bent according to potassium concentration and death time standard
Line, to be inferred to the death time of institute's test sample sheet.
Embodiment 6:
The detection method of the present embodiment is same as Example 3, only changes the DNA chain in embodiment into nucleotides sequence and is classified as core
The DNA chain of DNA sequence dna shown in nucleotide sequence table id NO.8, final measurement equally may be implemented to K+The quantitative survey of concentration
It is fixed, measure the vitreous humor K of different death times a series of+Concentration draws linear relationship chart, and potassium concentration and death is made
The standard curve of time measures the vitreous humor K to be measured of unknown death time+Concentration, according to potassium concentration and death time
Standard curve, to be inferred to the death time of institute's test sample sheet.
Embodiment 7:
The detection method of the present embodiment is same as Example 1, only changes the DNA chain in embodiment into nucleotides sequence and is classified as core
The DNA chain of DNA sequence dna shown in nucleotide sequence table id NO.9, final measurement equally may be implemented to K+The quantitative survey of concentration
It is fixed, measure the vitreous humor K of different death times a series of+Concentration draws linear relationship chart, and potassium concentration and death is made
The standard curve of time measures the vitreous humor K to be measured of unknown death time+Concentration, according to potassium concentration and death time
Standard curve, to be inferred to the death time of institute's test sample sheet.
Embodiment 8:
The detection method of the present embodiment is same as Example 2, only changes the DNA chain in embodiment into nucleotides sequence and is classified as core
DNA sequence dna shown in nucleotide sequence table id NO.5, final measurement equally may be implemented to K+The quantitative determination of concentration is surveyed
The vitreous humor K of fixed different death times a series of+Concentration draws linear relationship chart, and potassium concentration and death time is made
Standard curve, measure the vitreous humor K to be measured of unknown death time+Concentration, according to potassium concentration and death time standard
Curve, to be inferred to the death time of institute's test sample sheet.
Embodiment 9:
The detection method of the present embodiment is same as Example 3, only changes the DNA chain in embodiment into nucleotides sequence and is classified as core
DNA sequence dna shown in nucleotide sequence table id NO.13, final measurement equally may be implemented to K+The quantitative determination of concentration, it surveys
The vitreous humor K of fixed different death times a series of+Concentration draws linear relationship chart, and potassium concentration and death time is made
Standard curve, measure the vitreous humor K to be measured of unknown death time+Concentration, according to potassium concentration and death time standard
Curve, to be inferred to the death time of institute's test sample sheet.
SEQUENCE LISTING
<110>Central South University
Gorgeous monarch, fourth
River is insulted
Apricot plum, Lee
After peak, Cai
Yadong, Guo
Baiyi is drawn, Zha Laga
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Claims (6)
1. a kind of method of potassium concentration infering diing time in silver nanoclusters detection vitreous humor, which is characterized in that according to
Following steps carry out:
(1) it prepares fluorescent DNA-AgNCs: the DNA chain of AgNCs is designed containing potassium ion aptamers and can load first, in DNA chain
Middle addition AgNO3NaHB is added after oscillation in solution4Solution is put into refrigerator 3 hours after vibrating again, obtains brown color liquid, i.e., glimmering
Light DNA-AgNCs；It is described containing potassium ion aptamers and can to load the DNA chain of AgNCs be the DNA for being capable of forming the G- tetramer
Chain is selected from nucleotide sequence table id NO.1, nucleotide sequence table id NO.2, nucleotide sequence table id NO.3, nucleotide sequence
Table id NO.4, nucleotide sequence table id NO.5, nucleotide sequence table id NO.6, nucleotide sequence table id NO.7, nucleotide
Sequence table ID NO.8, nucleotide sequence table id NO.9, nucleotide sequence table id NO.10, nucleotide sequence table id NO.11,
DNA sequence dna shown in nucleotide sequence table id NO.12, nucleotide sequence table id NO.13；The DNA chain and AgNO3、NaHB4
Molar concentration rate is 1:2~10:2~10, preferably 1:6:6；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared: preparing various concentration potassium salt soln, is added into step
(1) in fluorescent DNA-AgNCs solution, its fluorescence intensity at 630nm is measured after reacting a period of time, it is dense to obtain potassium ion
The standard curve of degree and fluorescence intensity；
(3) actual glass body fluid sample potassium concentration detects: the vitreous humor to be measured of known death time is added to step
(1) in fluorescent DNA-AgNCs solution, its fluorescence intensity at 630nm is measured after reacting a period of time, according to preparation step
(2) potassium concentration and Standardization curve for fluorescence intensity obtains K in institute's test sample sheet+Concentration；
(4) potassium concentration and death time standard curve are prepared: according to the glass of known death time measured in step (3)
Body fluid K+The death time of concentration and known sample draws linear relationship chart, and the standard of potassium concentration and death time is made
Curve；
(5) corrupt corpse: the vitreous humor K to be measured of unknown death time is measured+Concentration, the potassium ion according to step (4) are dense
Degree and death time standard curve, to be inferred to the death time of institute's test sample sheet.
2. the side of potassium concentration infering diing time in a kind of silver nanoclusters detection vitreous humor according to claim 1
Method, which is characterized in that the pH value of the fluorescent DNA-AgNCs solution is 6-7.5.
3. the side of potassium concentration infering diing time in a kind of silver nanoclusters detection vitreous humor according to claim 1
Method, which is characterized in that the sylvite includes potassium chloride, potassium nitrate, potassium bromide and other potassium-containing compounds.
4. the side of potassium concentration infering diing time in a kind of silver nanoclusters detection vitreous humor according to claim 1
Method, which is characterized in that the solution to be measured is vitreous humor, blood, urine or other liquid of human or animal.
5. the side of potassium concentration infering diing time in a kind of silver nanoclusters detection vitreous humor according to claim 1
Method, which is characterized in that the reaction time of the step (2) and step (3) is 0.1~4 hour.
6. the side of potassium concentration infering diing time in a kind of silver nanoclusters detection vitreous humor according to claim 1
Method, which is characterized in that described dead for accurate supposition in legal medical expert's detection using potassium concentration in silver nanoclusters detection liquid
Time.
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| CN102866185A (en) * | 2012-09-06 | 2013-01-09 | 北京航空航天大学 | Method for detecting potassium ion by preparing biosensor based on G-quadruplex and gold nanoparticle |
| CN103913443A (en) * | 2014-04-23 | 2014-07-09 | 安徽师范大学 | Aptamer sensor based on DNA-Ag NCs (deoxyribonucleic acid-silver nanoclusters) as well as preparation method, application and detection method thereof |
| CN106018366A (en) * | 2016-05-09 | 2016-10-12 | 福建中医药大学 | Fluorescent DNA-silver nanocluster, and preparation method and application thereof |
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| CN102866185A (en) * | 2012-09-06 | 2013-01-09 | 北京航空航天大学 | Method for detecting potassium ion by preparing biosensor based on G-quadruplex and gold nanoparticle |
| CN103913443A (en) * | 2014-04-23 | 2014-07-09 | 安徽师范大学 | Aptamer sensor based on DNA-Ag NCs (deoxyribonucleic acid-silver nanoclusters) as well as preparation method, application and detection method thereof |
| CN106018366A (en) * | 2016-05-09 | 2016-10-12 | 福建中医药大学 | Fluorescent DNA-silver nanocluster, and preparation method and application thereof |
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| DNA-templated silver nanoclusters as laber-free sensitive detection probes for potassium ions and nitric oxide;Jihyun Lee等;《Journal of Materials Chemistry B》;20141231;第2卷;第2616-2621页 |
| 人体死后不同时间玻璃体液化学成分的变化趋势;陶涛等;《四川大学学报》;20061231;第37卷(第6期);第898-900页 |
| 人体玻璃体微量取样检测离子浓度推测死亡时间;贵州省遵义市公安局课题组;《侦查与技术》;20151231(第1期);第55-62页 |
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