The method for detecting acrylamide concentration in solution
Technical field
The invention belongs to technical field of chemical detection, more particularly to a kind of method for detecting acrylamide concentration in solution.
Background technology
Acrylamide is presented with mutagenesis with vitro test in vivo, can cause mammalian somatic cell and reproduction
The gene mutation of cell and chromosome abnormality, as micronucleus formation, Sister chromatid exohange, polyploid, aneuploid and other
Mitotic abnomality etc., dominant lethal test are positive.And prove that the metabolite glycidamide of acrylamide is that it is mainly caused
Mutagenic activity material.Animal experiment research finds that acrylamide can cause a variety of organ tumors of rat, including mammary gland, thyroid gland, testis
Ball, adrenal gland, nervous centralis, oral cavity, uterus, pituitary gland etc..International cancer research institution (IARC) 1994 is carcinogenic to its
Property is evaluated, and acrylamide is classified as into 2 class carcinogenic substances (2A) the i.e. mankind is potentially carcinogenic thing, and its Main Basis is acrylamide
It is metabolizable in animal and human body to be converted into its carcinogenic activity metabolite glycidamide.Therefore, the analysis of acrylamide
Detection is particularly important.So far, the detection method of acrylamide mainly has high performance liquid chromatography, liquid chromatogram coupling
Mass spectrography, fluorescent spectrometry etc..But these methods have, and pretreatment process is cumbersome, analysis time is long, instrument and cost of drugs height etc.
Deficiency.Therefore, establish simple, quick and high sensitivity acrylamide detection method and be increasingly becoming research emphasis.
After acrylamide enters in vivo, adduct can be combined to form with the guanine on DNA in vivo, cause gene mutation
Deng damage of genetic materials.Therefore acrylamide can be detected using electrochemical techniques using feature single stranded DNA as sensing platform,
So far, the relevant report that feature single stranded DNA modified glassy carbon electrode is used for acrylamide detection has not yet to see.
For problem above, we have studied a kind of glass carbon electricity modified based on carboxylated graphene and feature single stranded DNA
The new method of acrylamide is detected in pole, and this method is simple to operate, detection is quick and high sensitivity, can carry out the Gao Ling of acrylamide
Quick identification.
The content of the invention
As the result of various extensive and careful research and experiment, it has been found by the inventor that using carboxyl
Graphite alkene and the glass-carbon electrode of feature single stranded DNA modification, detect acrylamide using Differential Pulse Voltammetry, are favorably improved
The sensitivity of acrylamide detection.Based on this discovery, the present invention is completed.
It is an object of the invention to the method for the detection that have studied acrylamide concentration.
A further object of the invention is by using Differential Pulse Voltammetry, is used modified electrode as working electrode
Three-electrode system detects the concentration of acrylamide, establishes a kind of simple, quick and high sensitivity new side of acrylamide detection
Method.
A further object of the invention is that use is modified through carboxylated graphene and feature single stranded DNA in three-electrode system
Electrode be working electrode, the high sensitivity and the high-affinity of feature single stranded DNA that electrochemical signals conduct are combined by it.This
The three-electrode system of invention prepare it is easy, easily modification, it is sensitive it is high, testing expense is low, suitable for onlineization, stability it is good and with reference to mesh
Mark the advantages that thing scope is wide.
Technical scheme provided by the invention is:
A kind of method of acrylamide concentration in detection solution, including:
The acrylamide in liquid to be detected is detected by Differential Pulse Voltammetry using three-electrode system, according to third
The linear equation of acrylamide obtains the concentration of acrylamide in liquid to be detected, wherein, the working electrode in three-electrode system is carboxylic
Base graphite alkene and the glass-carbon electrode of feature single stranded DNA modification, the sequence of feature single stranded DNA is 5 '-AAA AAA AAA GGA
AAA AAA AA-(CH2)6-SH-3’。
Preferably, comprise the following steps in described detection solution in the method for acrylamide concentration:
Step 1: preparing the working electrode, the concentration for preparing acrylamide is 0-10nmol/L more parts of standard liquids;
Step 2: using the working electrode, it is molten to the every part of standard prepared in step 1 using Differential Pulse Voltammetry
Liquid is detected, and is obtained the differential pulse voltammetry volt-ampere curve of every part of standard liquid, is recorded every part of standard liquid respectively in the process
Differential pulse voltammetry volt-ampere curve in current strength peak value;
Step 3: with standard liquid corresponding to the differential pulse voltammetry volt-ampere curve of the every a standard liquid obtained in step 2
Standard liquid when being 0nmol/L of peak value and the acrylamide concentration of current strength current strength peak value difference divided by
The peak value of the current strength of this part of standard liquid is ordinate, and it is bent to draw standard using the concentration of every a standard liquid as abscissa
Line simultaneously calculates linear equation;
Step 4: using the working electrode, using Differential Pulse Voltammetry to the to be checked of the acrylamide of unknown concentration
Survey liquid to be detected, obtain the differential pulse voltammetry volt-ampere curve of liquid to be detected, the differential pulse voltammetry volt-ampere curve of liquid to be detected is corresponding
Liquid to be detected current strength peak value and acrylamide concentration be 0nmol/L standard liquid current strength peak value
The peak value of difference divided by the current strength of liquid to be detected is substituted into step 3 in obtained linear equation, and liquid to be detected is calculated
The concentration of middle acrylamide.
Preferably, in described detection solution in the method for acrylamide concentration, the step 1 specifically includes following
Step:
Step 1.1, the carboxylated graphene that 5 μ L are added dropwise on the glass-carbon electrode polished, dry under infrared lamp, wait to do
After dry, the glass-carbon electrode of carboxylated graphene modified is obtained, it is standby.
Step 1.2, the carboxylated graphene modified obtained in step 1.1 glass-carbon electrode on be added dropwise 5 μ L concentration for 1 ×
10-5The aqueous solution of mol/L feature single stranded DNA, and dried in 5 DEG C of refrigerator, after drying, obtain the carboxylated graphene
And the glass-carbon electrode of feature single stranded DNA modification;
Step 1.3, the acrylamide standard that the concentration of addition different volumes is 2 μm of ol/L into phosphate buffer solution are molten
Liquid, respectively obtain the solution a of the final concentration of 0nmol/L containing acrylamide, the final concentration of 1nmol/L containing acrylamide
Solution b, the solution c of final concentration of 2nmol/L containing acrylamide, final concentration of 5nmol/L containing acrylamide
Solution d, the solution a, solution b, solution c and solution d are four kinds of standard liquids;The pH of wherein described phosphate buffer solution is
7.0, concentration 0.2mol/L.
Preferably, the work is prepared in the step 1 in the method for acrylamide concentration in described detection solution
Before making electrode, in addition to the step of pre-processed to glass-carbon electrode, it is concretely comprised the following steps:
Glass-carbon electrode is placed sequentially in containing granularity is respectively 1 μm, 0.3 μm, on the polishing cloth of 0.05 μm of polishing powder
Polishing, the glass-carbon electrode is then placed sequentially in acetone, concentration is each super in 0.5mol/L sulfuric acid solution and ultra-pure water
Sound 1 minute, and ultrapure water 0.5min is used after each ultrasound terminates.
Preferably, in described detection solution in the method for acrylamide concentration, the step 2 specifically includes following
Step:
Step 2.1, three-electrode system is respectively implanted in four kinds of standard liquids, 3min is stirred at room temperature, it is static
3min;
Step 2.2, scanning differential pulse voltammetry collection of illustrative plates, it is 0V to set scanning initial potential, and termination current potential is 1.0V, current potential increment
For 0.004V, pulse width 0.05s, amplitude 0.05V, pulse period 0.5s, stand-by period 2s;
The peak value of step 2.3, the current strength for measuring respectively and recording four kinds of standard liquids, establish four kinds of marks
The differential pulse voltammetry volt-ampere curve of quasi- solution.
Preferably, in described detection solution in the method for acrylamide concentration, the reference in the three-electrode system
Electrode is Ag/AgCl electrodes, and auxiliary electrode is platinum electrode.
The present invention comprises at least following beneficial effect:
The present invention is using the glass that can be modified with the carboxylated graphene and feature single stranded DNA that acrylamide specificity is combined
Carbon electrode is working electrode, and the sensing interface for building correlation is used to detect as three-electrode system, improves detection acrylamide
Sensitivity, the minimum acrylamide that can detect 1nmol/L;
A kind of method of the present invention as the Electrochemical Detection acrylamide of feature based single stranded DNA, greatly reduces inspection
Cost is surveyed, it is simple to operate;
A step quick detection acrylamide of the invention, after the completion of prepared by three-electrode system, it is only necessary to which several minutes of cans are realized
One step detects acrylamide.
Further advantage, target and the feature of the present invention embodies part by following explanation, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
The term definition that the present invention relates to
Unless otherwise defined, otherwise all technologies used herein and scientific terminology all have with it is of the art
Those of ordinary skill generally understands identical implication.Although the usable and described herein in the practice or test of the present invention
Similar or equivalent any method, apparatus and material, but method for optimizing, device and material will now be described.
It is a kind of long-chain polymer that term " DNA ", which is, and composition unit is four kinds of deoxynucleotides, i.e. adenine deoxidation core
Thuja acid, thymidylic acid, deoxycytidylic acid, guanine deoxyribonucleoside acid.And deoxyribose and phosphoric acid point
Son is connected by ester bond, forms its long chain backbone, is arranged in outside, and four kinds of bases are arranged in inner side.Each glycan molecule and four
One of which in kind base is connected, and the sequence that these bases arrange along DNA long-chains, can form genetic code, refer to
Lead the synthesis of protein.The process for reading password is referred to as transcribing, and is to transcribe out one section so that one in DNA double chain is single-stranded for template
Referred to as mRNA nucleic acid molecules.
Term " single stranded DNA " refers to that double-stranded DNA is changed into single-stranded structure after heat or alkali process from double-spiral structure.It is single-stranded
DNA is different with double-stranded DNA in molecular fluid mechanical property, absorption spectrum, base reaction property etc..
" working electrode " is also known as Electrode, refers to that studied reaction occurs on this electrode.In general, to work
The basic demand of electrode is:Working electrode can be solid or liquid, and the conductive solid material of miscellaneous energy is equal
It can serve as electrode.The electrochemical reaction studied will not be affected because of the reaction that electrode itself is occurred, and can be
It is measured in larger potential areas;Electrode must not react with solvent or electrolyte component;Electrode area should not be too
Greatly, electrode surface preferably should be homogeneous smooth, and can carry out surface cleaning etc. by simple method.Using solid electrode
When, in order to ensure the reappearance of experiment, it has to be noted that suitable electrode pre-treatment step is established, to ensure redox, surface
Pattern and the reproducible state in the absence of adsorbing contaminant.In liquid electrode, mercury and amalgam are the most frequently used working electrodes, they
All it is liquid, there is reproducible equal phase surface, prepares and keep cleaning to be all easier to, while hydrogen high on electrode precipitation is super electric
The operation window note that gesture is improved under negative potential is widely used in electrochemical analysis.
Brief description of the drawings
Fig. 1 is the differential pulse voltammetry volt-ampere curve of acrylamide standard liquid of the present invention;
Fig. 2 is the standard curve of acrylamide of the present invention.
Embodiment
With reference to embodiment, the present invention is described in further detail, to make those skilled in the art with reference to specification
Word can be implemented according to this.
Embodiment 1
The present invention provides a kind of method for detecting acrylamide concentration in solution, including:
The acrylamide in liquid to be detected is detected by Differential Pulse Voltammetry using three-electrode system, according to third
The linear equation of acrylamide obtains the concentration of acrylamide in liquid to be detected, wherein, the working electrode in three-electrode system is carboxylic
Base graphite alkene and the glass-carbon electrode of feature single stranded DNA modification, the sequence of feature single stranded DNA is 5 '-AAA AAA AAA GGA
AAA AAA AA-(CH2)6-SH-3’.Three-electrode system is made up of working electrode, reference electrode and auxiliary electrode.
The present invention is that the acrylamide in liquid to be detected is detected using Differential Pulse Voltammetry, differential pulse voltammetry volt-ampere
Method is superimposed the continuous impulse that an amplitude constant and pulsewidth are fixed on linear sweep waveform, and in scanning process, base current potential is from initial
Electric potential scanning is carrying out current sample, by the two sample rate currents before potential pulse starts to current potential is terminated with the end of
Difference is mapped to current potential, as differential pulse voltammetry volt-ampere curve, is mainly used in electrochemical analysis, is reduced because of oxidation of impurities reduction reaction
Caused background current, there is more preferable detection sensitivity and lower detectable limit, relative to other method, differential pulse voltammetry volt
Peace method cost is relatively low, simple to operate.
Present invention employs following mechanism:Graphene can be combined with glass-carbon electrode by π-pi-conjugated principle, guanine energy
Acted on acrylamide, both combinations can change guanine oxidizing intensity, and guanine is tested using Differential Pulse Voltammetry
The change of oxidation signal can be used for detecting the concentration of detection acrylamide.Contain the spy of sulfydryl in one section of 3 ' end of present invention design
Levy single stranded DNA, with graphene and DNA electrostatic interaction, using method of modifying is added dropwise, make feature single stranded DNA successfully modify in
On glass-carbon electrode, because containing guanine in feature single stranded DNA, carrying out blank solution, (i.e. acrylamide concentration is 0nmol/L's
Solution) test when, there are strong electrochemical signals, but when acrylamide solution is added after, acrylamide and guanine effect, make
Obtain electrochemical signals to change, the change by signal is the concentration of measurable acrylamide.
In described detection solution in the method for acrylamide concentration, comprise the following steps:
Step 1: preparing the working electrode, the concentration for preparing acrylamide is 0-10nmol/L more parts of standard liquids;
Step 2: using the working electrode, it is molten to the every part of standard prepared in step 1 using Differential Pulse Voltammetry
Liquid is detected, and is obtained the differential pulse voltammetry volt-ampere curve of every part of standard liquid, is recorded every part of standard liquid respectively in the process
Differential pulse voltammetry volt-ampere curve in current strength peak value;
Step 3: with standard liquid corresponding to the differential pulse voltammetry volt-ampere curve of the every a standard liquid obtained in step 2
Standard liquid when being 0nmol/L of peak value and the acrylamide concentration of current strength current strength peak value difference divided by
The peak value of the current strength of this part of standard liquid is ordinate, and it is bent to draw standard using the concentration of every a standard liquid as abscissa
Line simultaneously calculates linear equation;
Step 4: using the working electrode, using Differential Pulse Voltammetry to the to be checked of the acrylamide of unknown concentration
Survey liquid to be detected, obtain the differential pulse voltammetry volt-ampere curve of liquid to be detected, the differential pulse voltammetry volt-ampere curve of liquid to be detected is corresponding
Liquid to be detected current strength peak value and acrylamide concentration be 0nmol/L standard liquid current strength peak value
The peak value of difference divided by the current strength of liquid to be detected is substituted into step 3 in obtained linear equation, and liquid to be detected is calculated
The concentration of middle acrylamide.
In described detection solution in the method for acrylamide concentration, the step 1 specifically includes following steps:
Step 1.1, the carboxylated graphene that 5 μ L are added dropwise on the glass-carbon electrode polished, dry under infrared lamp, wait to do
After dry, the glass-carbon electrode of carboxylated graphene modified is obtained, it is standby.
Step 1.2, the carboxylated graphene modified obtained in step 1.1 glass-carbon electrode on be added dropwise 5 μ L concentration for 1 ×
10-5The aqueous solution of mol/L feature single stranded DNA, and dried in 5 DEG C of refrigerator, after drying, obtain the carboxylated graphene
And the glass-carbon electrode of feature single stranded DNA modification;
Step 1.3, the acrylamide standard that the concentration of addition different volumes is 2 μm of ol/L into phosphate buffer solution are molten
Liquid, respectively obtain the solution a of the final concentration of 0nmol/L containing acrylamide, the final concentration of 1nmol/L containing acrylamide
Solution b, the solution c of final concentration of 2nmol/L containing acrylamide, final concentration of 5nmol/L containing acrylamide
Solution d, the solution a, solution b, solution c and solution d are four kinds of standard liquids;The pH of wherein described phosphate buffer solution is
7.0, concentration 0.2mol/L.
In described detection solution in the method for acrylamide concentration, prepared in the step 1 working electrode it
Before, in addition to the step of pre-processed to glass-carbon electrode, it is concretely comprised the following steps:
Glass-carbon electrode is placed sequentially in containing granularity is respectively 1 μm, 0.3 μm, on the polishing cloth of 0.05 μm of polishing powder
Polishing, the glass-carbon electrode is then placed sequentially in acetone, concentration is each super in 0.5mol/L sulfuric acid solution and ultra-pure water
Sound 1 minute, and ultrapure water 0.5min is used after each ultrasound terminates.
In described detection solution in the method for acrylamide concentration, the step 2 specifically includes following steps:
Step 2.1, three-electrode system is respectively implanted in four kinds of standard liquids, 3min is stirred at room temperature, it is static
3min;
Step 2.2, scanning differential pulse voltammetry collection of illustrative plates, it is 0V to set scanning initial potential, and termination current potential is 1.0V, current potential increment
For 0.004V, pulse width 0.05s, amplitude 0.05V, pulse period 0.5s, stand-by period 2s;
The peak value of step 2.3, the current strength for measuring respectively and recording four kinds of standard liquids, establish four kinds of marks
The differential pulse voltammetry volt-ampere curve of quasi- solution.The differential pulse voltammetry volt-ampere curve of four kinds of standard liquids is as shown in Figure 1.Wherein curve a, b,
C, d is respectively solution a, solution b, the differential pulse voltammetry volt-ampere curve of tetra- kinds of standard liquids of solution c and solution d;
Respectively with the peak value and acrylamide concentration of current strength corresponding to solution a, b, c, d differential pulse voltammetry volt-ampere curve
For 0nmol/L when standard liquid current strength the difference of peak value divided by the peak value of current strength of this part of standard liquid be
Ordinate, standard curve is drawn as abscissa using the concentration of acrylamide in solution a, b, c, d and calculates linear equation;For example,
Standard during using the peak value of current strength corresponding to solution a differential pulse voltammetry volt-ampere curve and acrylamide concentration as 0nmol/L
The peak value of the difference of the peak value of the current strength of solution divided by solution a current strength is ordinate, with acrylamide in solution a
Concentration be abscissa, it may be determined that a point on standard curve, by the above method, change solution a into solution b, c, d,
The other three point on standard curve is determined by solution b, c, d, draws standard curve as shown in Figure 2.
Linear equation is obtained by standard curve:Y=0.066+0.38X, Y is current value (I-I in formula0)/I, I are every part of mark
The peak value of current strength corresponding to the differential pulse voltammetry volt-ampere curve of quasi- solution;I0Mark when for acrylamide concentration being 0nmol/L
The peak value of the current strength of quasi- solution, unit are μ A;X be standard liquid in acrylamide concentration, unit nmol/L, phase relation
Number R2For 0.992.Detection of the modified electrode to acrylamide is limited to 1nmol/L.
Calculate the concentration of acrylamide in liquid to be detected:Will be to be detected corresponding to the differential pulse voltammetry volt-ampere curve of liquid to be detected
The difference of the peak value of the current strength of liquid and the peak value of the current strength of standard liquid that acrylamide concentration is 0nmol/L divided by
The peak value of the current strength of liquid to be detected is substituted into linear equation as Y value, and the dense of acrylamide in liquid to be detected is calculated
Degree.
In described detection solution in the method for acrylamide concentration, the reference electrode in the three-electrode system is Ag/
AgCl electrodes, auxiliary electrode are platinum electrode.
Embodiment 2
Electrochemical workstation:CHI760E;
Three-electrode system:Working electrode is the glass-carbon electrode modified through carboxylated graphene and feature single stranded DNA, feature list
The sequence of chain DNA is 5 '-AAA AAA AAA GGA AAA AAA AA- (CH2)6- SH-3 ', its 3 ' end section modification sulfydryl;
Auxiliary electrode is platinum electrode;Reference electrode is Ag/AgCl electrodes;
Cushioning liquid:PH value is 7.0, and concentration is 0.2mol/L phosphate buffer solution;
Standard Stock solutions:Concentration is 2 μm of ol/L acrylamide standard liquid;
Processing of the glass-carbon electrode before modification:Glass-carbon electrode is placed sequentially in containing granularity is respectively 1 μm, 0.3 μm,
Polished on the polishing cloth of 0.05 μm of polishing powder, the glass-carbon electrode is then placed sequentially in acetone, concentration 0.5mol/L
Sulfuric acid solution and ultra-pure water in each ultrasonic 1 minute, and ultrapure water 0.5min is used after each ultrasound terminates.
The modification of glass-carbon electrode:5 μ L carboxylated graphene is added dropwise on the glass-carbon electrode polished, is done under infrared lamp
It is dry, after to be dried, the glass-carbon electrode of carboxylated graphene modified is obtained, is added dropwise on the glass-carbon electrode of carboxylated graphene modified
5 μ L concentration are 1 × 10-5The aqueous solution of mol/L feature single stranded DNA, and dried in 5 DEG C of refrigerator, after drying, obtain described
Carboxylated graphene and the glass-carbon electrode of feature single stranded DNA modification.
Assay method:The acrylamide standard that the concentration that different volumes are added into phosphate buffer solution is 2 μm of ol/L is molten
Liquid, respectively obtain the solution a of the final concentration of 0nmol/L containing acrylamide, the final concentration of 1nmol/L containing acrylamide
Solution b, the solution c of final concentration of 2nmol/L containing acrylamide, final concentration of 5nmol/L containing acrylamide
Solution d, the solution a, solution b, solution c and solution d are four kinds of standard liquids.
The three-electrode system is respectively implanted in four kinds of titers, 3min, static 3min is stirred at room temperature;Sweep
Retouch differential pulse voltammetry collection of illustrative plates, it is 0V to set scanning initial potential, and termination current potential is 0.7V, and current potential increment is 0.004V, differential pulse voltammetry
Frequency is 50Hz, amplitude 0.05V, stand-by period 2s;Measure respectively and record the current strength of four kinds of standard liquids
Peak value, establish the differential pulse voltammetry volt-ampere curves of four kinds of standard liquids.The differential pulse voltammetry volt-ampere curve of four kinds of standard liquids
As shown in figure 1, wherein curve a, b, c, d is respectively solution a, solution b, the differential pulse voltammetry of tetra- kinds of standard liquids of solution c and solution d
Volt-ampere curve.
Respectively with the peak value and acrylamide concentration of current strength corresponding to solution a, b, c, d differential pulse voltammetry volt-ampere curve
For 0nmol/L when standard liquid current strength the difference of peak value divided by the peak value of current strength of this part of standard liquid be
Ordinate, standard curve is drawn as abscissa using the concentration of acrylamide in solution a, b, c, d and calculates linear equation;Example
Mark when such as, using the peak value of current strength corresponding to solution a differential pulse voltammetry volt-ampere curve and acrylamide concentration as 0nmol/L
The peak value of the difference of the peak value of the current strength of quasi- solution divided by solution a current strength is ordinate, with acryloyl in solution a
The concentration of amine is abscissa, it may be determined that a point on standard curve, similarly, by the above method, changes solution a into solution
B, c, d, the other three point on standard curve is determined by solution b, c, d.Draw standard curve as shown in Figure 2.
Linear equation is obtained by standard curve:Y=0.066+0.38X, Y is current value (I-I in formula0)/I, I are every part of mark
The peak value of current strength corresponding to the differential pulse voltammetry volt-ampere curve of quasi- solution;I0Standard when acrylamide concentration is 0nmol/L
The peak value of the current strength of solution, unit are μ A;X be standard liquid in acrylamide concentration, unit nmol/L, coefficient correlation
R2For 0.992.Detection of the modified electrode to acrylamide is limited to 1nmol/L.
The measure of liquid to be detected:The three-electrode system is immersed in the 10mL phosphate buffer solution, described
1mL is added in phosphate buffer and contains the acrylamide solution of unknown concentration, stirs 3min, static 3min, as liquid to be detected,
The differential pulse voltammetry collection of illustrative plates of liquid to be detected is scanned, it is 0V to set scanning initial potential, and termination current potential is 1.0V, and current potential increment is
0.004V, pulse width 0.05s, amplitude 0.05V, pulse period 0.5s, stand-by period 2s, sensitivity 10-6A/V, obtain
To the differential pulse voltammetry volt-ampere curve of liquid to be detected, measure and record the peak value of the current strength of liquid to be detected.
By the peak value and acrylamide of the current strength of liquid to be detected corresponding to the differential pulse voltammetry volt-ampere curve of liquid to be detected
Concentration be 0nmol/L standard liquid current strength peak value difference divided by liquid to be detected current strength peak value conduct
Y value is substituted into linear equation, and the concentration of acrylamide in liquid to be detected is calculated.
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited
In specific details and shown here as the embodiment with description.