CN104198565B - A kind of quantitative detection of folic acid device and preparation method thereof and its application in quantitative detection of folic acid - Google Patents
A kind of quantitative detection of folic acid device and preparation method thereof and its application in quantitative detection of folic acid Download PDFInfo
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- CN104198565B CN104198565B CN201410335047.3A CN201410335047A CN104198565B CN 104198565 B CN104198565 B CN 104198565B CN 201410335047 A CN201410335047 A CN 201410335047A CN 104198565 B CN104198565 B CN 104198565B
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- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 title claims abstract description 515
- 235000019152 folic acid Nutrition 0.000 title claims abstract description 270
- 239000011724 folic acid Substances 0.000 title claims abstract description 270
- 229960000304 Folic Acid Drugs 0.000 title claims abstract description 267
- 238000001514 detection method Methods 0.000 title claims abstract description 95
- 238000002360 preparation method Methods 0.000 title claims abstract description 37
- 150000002484 inorganic compounds Chemical class 0.000 claims abstract description 37
- 229910010272 inorganic material Inorganic materials 0.000 claims abstract description 37
- 150000002894 organic compounds Chemical class 0.000 claims abstract description 37
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims abstract description 34
- 239000008346 aqueous phase Substances 0.000 claims abstract description 33
- 239000012071 phase Substances 0.000 claims abstract description 27
- 239000011259 mixed solution Substances 0.000 claims abstract description 26
- 229910052697 platinum Inorganic materials 0.000 claims abstract description 16
- 230000035945 sensitivity Effects 0.000 claims abstract description 15
- HKZLPVFGJNLROG-UHFFFAOYSA-M Silver chloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 claims abstract description 14
- 239000012488 sample solution Substances 0.000 claims description 50
- 239000012086 standard solution Substances 0.000 claims description 50
- FAPWRFPIFSIZLT-UHFFFAOYSA-M sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 35
- 239000007853 buffer solution Substances 0.000 claims description 31
- 239000000243 solution Substances 0.000 claims description 30
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 27
- 239000003115 supporting electrolyte Substances 0.000 claims description 26
- 239000007864 aqueous solution Substances 0.000 claims description 23
- 230000000875 corresponding Effects 0.000 claims description 20
- 239000011780 sodium chloride Substances 0.000 claims description 20
- NGVCICDLVFASRT-UHFFFAOYSA-N C1(=CC=CC=C1)P(C1=CC=CC=C1)C1=CC=CC=C1.[Cl-].[NH4+] Chemical compound C1(=CC=CC=C1)P(C1=CC=CC=C1)C1=CC=CC=C1.[Cl-].[NH4+] NGVCICDLVFASRT-UHFFFAOYSA-N 0.000 claims description 16
- VEXZGXHMUGYJMC-UHFFFAOYSA-M chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 16
- 238000003950 stripping voltammetry Methods 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 15
- OVISMSJCKCDOPU-UHFFFAOYSA-N 1,6-dichlorohexane Chemical compound ClCCCCCCCl OVISMSJCKCDOPU-UHFFFAOYSA-N 0.000 claims description 14
- 238000004062 sedimentation Methods 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- SCJNXSCHPUAAGK-UHFFFAOYSA-N B(O)(O)O.ClC1=CC=C(C=C1)[K] Chemical compound B(O)(O)O.ClC1=CC=C(C=C1)[K] SCJNXSCHPUAAGK-UHFFFAOYSA-N 0.000 claims description 10
- 239000012528 membrane Substances 0.000 claims description 10
- GNSKLFRGEWLPPA-UHFFFAOYSA-M Monopotassium phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 9
- 238000000227 grinding Methods 0.000 claims description 9
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 9
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 238000005070 sampling Methods 0.000 claims description 8
- KKEYFWRCBNTPAC-UHFFFAOYSA-L terephthalate(2-) Chemical compound [O-]C(=O)C1=CC=C(C([O-])=O)C=C1 KKEYFWRCBNTPAC-UHFFFAOYSA-L 0.000 claims description 8
- 238000011068 load Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 229960000935 Dehydrated Alcohol Drugs 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N HCl Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- 229910000041 hydrogen chloride Inorganic materials 0.000 claims description 6
- OZAIFHULBGXAKX-UHFFFAOYSA-N precursor Substances N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 claims description 6
- 239000000376 reactant Substances 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 235000012239 silicon dioxide Nutrition 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 5
- 238000004090 dissolution Methods 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 239000002244 precipitate Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 238000007792 addition Methods 0.000 claims description 4
- RMAQACBXLXPBSY-UHFFFAOYSA-N Silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 150000002148 esters Chemical class 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 229910052904 quartz Inorganic materials 0.000 claims description 3
- 239000010453 quartz Substances 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims description 3
- 229910002027 silica gel Inorganic materials 0.000 claims description 3
- 238000010521 absorption reaction Methods 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N Triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 claims 6
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims 3
- 235000019270 ammonium chloride Nutrition 0.000 claims 3
- QDHHCQZDFGDHMP-UHFFFAOYSA-N monochloramine Chemical compound ClN QDHHCQZDFGDHMP-UHFFFAOYSA-N 0.000 claims 3
- 230000004044 response Effects 0.000 abstract description 26
- -1 Folic Acid anion Chemical class 0.000 description 19
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- 229940079593 drugs Drugs 0.000 description 4
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- 206010034695 Pernicious anaemia Diseases 0.000 description 1
- HNXQXTQTPAJEJL-UHFFFAOYSA-N Pterin Chemical class C1=CN=C2NC(N)=NC(=O)C2=N1 HNXQXTQTPAJEJL-UHFFFAOYSA-N 0.000 description 1
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 description 1
- 210000003462 Veins Anatomy 0.000 description 1
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Abstract
The present invention discloses device and the preparation method and application of a kind of quantitative detection of folic acid, and the device of described quantitative detection of folic acid includes aqueous phase pond and oil phase pond；Described aqueous phase pond includes that the container of upper end open, aqueous phase platinum filament are to electrode II, aqueous phase reference electrode Ag/AgCl;Described oil phase pond includes that organogel electrochemical sensor, oil phase platinum filament are to electrode I, oil phase reference electrode Ag/AgCl；Described organogel electrochemical sensor is that bottom is stained with MCM 41 type organic/inorganic compound mesoporous film and bottom-up in the hollow pipe of upper opening, and hollow pipe is sequentially arranged with organogel, mixed solution.Preparation method includes 4 steps such as the device assembling of MCM 41 type organic/inorganic compound mesoporous film, organogel, the preparation of organogel electrochemical sensor and quantitative detection of folic acid.The method utilizing this device that folic acid carries out detection by quantitative has the highest sensitivity, precision, accuracy, the response rate and repeatability.
Description
Technical field
The present invention relates to a kind of quantitative detection of folic acid device and preparation method thereof and its application in quantitative detection of folic acid, belong to liquid/liquid interface electrochemical field and electrochemical membrane sensor device application.
Background technology
Folic acid (folic acid, FA) is pterin derivatives, belongs to water-soluble B vitamin.Also known as pteroylglutamic acid, is body cell growth and the required material of breeding, and its molecular formula is C19H19N7O6, molecular weight is 441.40.The Mental Health of human body is played an important role by folic acid, and as main one carbon unit donor, folic acid participates in nucleic acid, aminoacid, the biosynthesis of protein.Folic acid is mainly used in folic acid deficiency that a variety of causes causes and the treatment of the megaloblastic anemia that folic acid deficiency causes, the auxiliary treatment of pernicious anemia, and the prevention of trimester of pregnancy, women breast-feeding their children are administered.Owing to folic acid has these special physiological functions, particularly to trimester of pregnancy, women breast-feeding their children and fetus, folic acid is through being used in health care medicine frequently as nutrition enhancer, therefore, how to measure the folate content in these medicines, just become particularly important monitoring the quality of these medicines.
At present, the apparatus and method about folic acid detection are the most more complicated both at home and abroad, are broadly divided into electrochemical method and non-electrochemical method.Non-electrochemical method typically has microbial method, high performance liquid chromatography, euzymelinked immunosorbent assay (ELISA) method and fluorescence spectrophotometry.Microbial method complex operation, easily causes cross infection, and repeatability is bad.High performance liquid chromatography not only expensive equipment but also pipeline easily corrode, the long-term damage measuring the equipment that easily causes.Euzymelinked immunosorbent assay (ELISA) testing result is quite different with actual numerical value, and life-time service is relatively costly.The fluorescence spectrophotometry response rate is less than prescribed limit, it is impossible to for actually detected.Electrochemical method focuses primarily upon the solid liquid interface electrochemistry utilizing modified electrode, but these modified electrode manufacturing process often relate to some hypertoxicity material or precious metals, and modified electrode is used for multiple times and is easily lost, have and be unfavorable for health, relatively costly and the shortcoming such as can not reuse.Based on the problems referred to above, develop a kind of prepare simple, operate quantitative detection of folic acid device easy and with low cost, utilize this quantitative detection of folic acid device the most particularly important for quantitative detection of folic acid result favorable reproducibility, the folic acid detection method that highly sensitive, the response rate is high simultaneously.
Summary of the invention
An object of the present invention be in order to solve complex operation in the presence of prior art Folic Acid quantitative testing device, be easily damaged, the technical problem such as relatively costly and a kind of quantitative detection of folic acid device is provided, this device has that preparation is simple, operate the characteristics such as easy and with low cost.
The two of the purpose of the present invention are to provide the preparation method of above-mentioned a kind of quantitative detection of folic acid device.
The three of the purpose of the present invention are to provide a kind of method utilizing above-mentioned quantitative detection of folic acid device to carry out quantitative detection of folic acid, and this detection method has the highest sensitivity, precision, accuracy and the response rate and service life is long, favorable reproducibility.
The know-why of the present invention
The liquid/liquid interface electrochemistry that perforated membrane supports has mass transfer enhancement because of it, expands electrochemical signals and improves the advantages such as Electroanalytical Chemistry sensitivity, is widely used in biomacromolecule and drug molecule detection in recent years.On liquid/liquid interface, organogel combines the use of Stripping Voltammetry technology so that the detection limit of some biomacromolecules and drug molecule is significantly reduced to sub-micromolar rank.The liquid/liquid interface electrochemical method detection folic acid supported according to perforated membrane.For problems of the prior art, utilize and containing the MCM-41 type organic/inorganic compound mesoporous film of ionic surfactant (CTAB), anion is had ion exchanging function and combine the preenrichment function of organogel and substantially increase the detection limit of folic acid, it is achieved that the trace analysis of folic acid and detection.
Technical scheme
The device of a kind of quantitative detection of folic acid, including aqueous phase pond and oil phase pond；Oil phase pond is placed in the middle and upper part in aqueous phase pond, and aqueous phase pond can be moved up and down by lowering or hoisting gear；
Described aqueous phase pond includes that the container of upper end open, aqueous phase platinum filament are to electrode II, aqueous phase reference electrode Ag/AgCl;
Described oil phase pond includes that organogel electrochemical sensor, oil phase platinum filament are to electrode I, oil phase reference electrode Ag/AgCl；
Described organogel electrochemical sensor is that bottom is stained with MCM-41 type organic/inorganic compound mesoporous film and bottom-up in the hollow pipe of upper opening, and hollow pipe is sequentially arranged with organogel, mixed solution；
The material of the hollow pipe of described upper opening is quartz；
The volume of organogel accounts for the 10-20% of whole hollow pipe volume, and the volume of mixed solution accounts for the 30-80% of hollow pipe volume;
Described mixed solution be concentration be 10mol L-1Double (triphenylphosphine) ammonium chloride) aqueous solution and concentration is 0.1mol L-1NaCl aqueous solution calculate by volume, i.e. 10mol L-1Double (triphenylphosphine) ammonium chloride) aqueous solution: 0.1mol L-1 The ratio that NaCl aqueous solution is 5:6 mix；
Described organogel method as follows is prepared from:
By 1,6-dichloro hexane and organic facies supporting electrolyte heat 10-50min in 40-60 DEG C of oil bath, it is subsequently adding the polrvinyl chloride that mean molecule quantity is 22000, it is to slowly warm up to 70-90 DEG C, after the transparent state of solution, control temperature and be 65-85 DEG C, constant temperature stirring 10-50min, obtain organogel；
Wherein 1,6-dichloro hexane, organic facies supporting electrolyte and the consumption of polrvinyl chloride that mean molecule quantity is 22000, by 1,6-dichloro hexane: organic facies supporting electrolyte: mean molecule quantity is the ratio that polrvinyl chloride is 1ml:0.012g:0.02-0.5g of 22000；
Organic facies supporting electrolyte wherein used is prepared by a method comprising the following steps and forms:
Double (triphenylphosphine) ammonium chloride and four (4-chlorphenyl) potassium borate are dissolved in respectively the mixed solution formed for the first alcohol and water of 1:1 by volume, again by two kinds of solution mixing, until there being white precipitate to generate, i.e. prepare organic facies supporting electrolyte, the molar concentration of the most double (triphenylphosphine) ammonium chloride is 0.01mol/L, and the molar concentration of four (4-chlorphenyl) potassium borate is 0.01mol/L.
The preparation of the device of above-mentioned a kind of quantitative detection of folic acid, specifically includes following steps:
(1), the preparation of MCM-41 type organic/inorganic compound mesoporous film
By dehydrated alcohol, positive silicic acid four ester (TEOS) and 2.8mmol/L HCl/water solution in the ratio mix homogeneously of 1g:1-5g:0.1-1ml, put into there-necked flask, being placed in oil bath, temperature all controls at 50-80 DEG C, reacts 90min, stopping heating, the reactant liquor of gained is cooled to room temperature；
By cetyl trimethylammonium bromide (CTAB), dehydrated alcohol and 55mmol/L
HCl/water solution joins after mixing in the ratio of 1g:5-15g:1-5ml in the reactant liquor of above-mentioned gained and continues stirring 10-50min, obtain precursor liquid, then, under the conditions of room temperature 25 DEG C, blank for precursor liquid obtained by 2mL terephthalate fat (PET) film is filtered in the way of sucking filtration, keep sucking filtration state 2min, after the substantially dry of i.e. blank terephthalate fat (PET) film surface, obtain MCM-41 type organic/inorganic compound mesoporous film, be specifically shown in granted patent ZL 2,011 1
Shown in 0098700.5；
(2), the preparation of organogel
By 1,6-dichloro hexane and organic facies supporting electrolyte heat 10-50min in 40-60 DEG C of oil bath, it is subsequently adding the polrvinyl chloride that mean molecule quantity is 22000, it is to slowly warm up to 70-90 DEG C, after the transparent state of solution, control temperature and be 65-85 DEG C, constant temperature stirring 10-50min, obtain organogel；
Wherein 1,6-dichloro hexane, organic facies supporting electrolyte and the consumption of polrvinyl chloride that mean molecule quantity is 22000, by 1,6-dichloro hexane: organic facies supporting electrolyte: mean molecule quantity is the ratio that polrvinyl chloride is 5ml:0.059g:0.1-1.0g of 22000；
Organic facies supporting electrolyte wherein used is prepared by a method comprising the following steps and forms:
Double (triphenylphosphine) ammonium chloride and four (4-chlorphenyl) potassium borate are dissolved in respectively the mixed solution formed for the first alcohol and water of 1:1 by volume, again by two kinds of solution mixing, until there being white precipitate to generate, i.e. prepare organic facies supporting electrolyte, the molar concentration of the most double (triphenylphosphine) ammonium chloride is 0.01mol/L, and the molar concentration of four (4-chlorphenyl) potassium borate is 0.01mol/L；
(3), the preparation of organogel electrochemical sensor
With silica gel, the MCM-41 type organic/inorganic compound mesoporous film of step (1) gained is bonded at the opening of the channel bottom a hollow pipe, places at least 12h；
Being injected into bottom with glass dropper absorption organogel again to be stained with in the hollow pipe of MCM-41 type organic/inorganic compound mesoporous film, place at least 1h, in order to make organogel harden, the volume of organogel accounts for the 10-20% of whole hollow pipe volume;
After the organogel in hollow pipe hardens, being added thereto by mixed solution, its addition accounts for the 30-80% of hollow pipe volume, obtains organogel electrochemical sensor；
Described mixed solution is in 50ml volumetric flask, and adding 5ml concentration is 10mol L-1Double (triphenylphosphine) ammonium chloride) aqueous solution and 6ml concentration is 0.1mol L-1NaCl aqueous solution after be settled to 50ml with distilled water；
(4), the assembling of the device of quantitative detection of folic acid
Fixing device is utilized to be fixed on the container of a upper end open by the organogel electrochemical sensor of step (3) gained in shielded box, one end with MCM-41 type organic/inorganic compound mesoporous film is downward, and the position with MCM-41 type organic/inorganic compound mesoporous film is positioned at the middle and upper part of container, finally, oil phase platinum filament is inserted to electrode I in the mixed solution of the hollow pipe of organogel electrochemical sensor, oil phase reference electrode Ag/AgCl, aqueous phase platinum filament is inserted to electrode II in the container of upper end open, aqueous phase pond, aqueous phase reference electrode Ag/AgCl, i.e. complete the assembling of the device of quantitative detection of folic acid.
The device utilizing above-mentioned quantitative detection of folic acid carries out the detection of folic acid, specifically includes following steps:
(1), the preparation of folic acid standard solution
Compound concentration is respectively 1 μM/L, 3 μMs/L, 5 μMs/L, 9 μMs/L, 18 μMs/L, 24 μMs/L, 32 μMs/L, the folic acid standard solution of 40 μMs/L；
The solvent used by preparation of described folic acid standard solution is pH=7.2, containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and the aqueous solution of 0.1mol/L sodium hydroxide, its configuration process will be dissolved in above-mentioned solvent by folic acid, then constant volume is in 50ml volumetric flask, obtain the folic acid standard solution of variable concentrations, be put in darkroom and save backup；
(2) preparation of folic acid sample solution, to be measured
Testing sample is as a example by YESUAN PIAN, take 1 YESUAN PIAN and put into grinding alms bowl, grind with grinding rod, load 25ml small beaker, 15ml buffer solution 1 ultrasonic dissolution 10min is added in beaker, then 25ml centrifuge tube is proceeded to, closeing the lid, put into centrifuge, revolution is 10000-25000 × g, centrifugal 20-50min, described supernatant proceeds to 50ml volumetric flask buffer solution 2 and is settled to 50ml, then pipettes 5ml with pipet, proceeds to 50ml volumetric flask buffer solution 3 and is settled to 50ml, obtain folic acid sample solution to be measured, put into darkroom and save backup；
Described buffer solution 1, buffer solution 2, buffer solution 3 are pH=7.2, containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and the aqueous solution of 0.1mol/L sodium hydroxide；
(3), utilize the device of quantitative detection of folic acid, measure the peak point current corresponding to folic acid standard solution and folic acid sample solution to be measured by electrochemical workstation；
By in the container loading the upper end open of the device of quantitative detection of folic acid at folic acid sample solution to be measured or folic acid standard solution, by small lifting table device, the container of upper end open is little by little risen, until folic acid standard solution or folic acid sample solution to be measured by chance touch the MCM-41 type organic/inorganic compound mesoporous film bottom the hollow pipe of organogel electrochemical sensor, operating process can not make the MCM-41 type organic/inorganic compound mesoporous film bottom hollow pipe be immersed in folic acid standard solution or folic acid sample solution to be measured;
Utilize electrochemical workstation, with middle cyclic voltammetry (CV), at its each technical parameter such as high potential: 0.8-1.2V, electronegative potential: 0-0.2V, sweep speed: 0.005-0.1V/s, scan hop count: 2-10, sensitivity: 10-6-10-4Under conditions of and differential pulse voltammetry (DPV), at its each technical parameter such as initial potential: 0-0.2V, termination current potential: 0.8-1.2V, pulse width: 0.01-0.1V, sampling width: 0.01-0.1s, pulse period: 0.1-0.5s, sensitivity: 10-6-10-4Under conditions of observe folic acid solution Folic Acid anion electrochemical response on this organogel electrochemical sensor;
With the preenrichment function of Differential Pulse Stripping Voltammetry (DPSV), at each technical parameter such as amplitude: 0.01-0.1V, pulse width: 0.01-0.1V, sampling width: 0.01-0.1s, pulse period: 0.1-0.5s, sedimentation potential: 0.1-0.8V, sedimentation time: 5-100s, sensitivity: 10-6-10-4Under conditions of the peak current corresponding to folic acid sample solution respectively to be measured in read step (1) Folic Acid standard solution and step (2);
(4), the drafting of standard curve；
With peak current as vertical coordinate, folic acid concentration corresponding to folic acid standard solution is abscissa, draws the curve between the folic acid concentration corresponding to peak current and folic acid standard solution, and matching obtains the standard curve equation of folic acid concentration and peak current；
(5), folic acid sample Folic Acid concentration to be measured
The peak current of folic acid sample solution to be measured in step (3) is substituted in step (4) the gained folic acid concentration standard curve equation with peak current, and it is calculated the concentration of folic acid sample solution Folic Acid to be measured, then according to the extension rate of step (2) folic acid to be measured sample, thus the content of folic acid sample Folic Acid to be measured is obtained.
Beneficial effects of the present invention
A kind of device of the quantitative detection of folic acid of the present invention, folic acid detection device complex operation, the shortcomings such as repeatability is bad, and use cost is higher, and the response rate is low before overcoming.Wherein organogel electrochemical sensor has that preparation is simple, operate the characteristics such as easy and with low cost.The device agent of this quantitative detection of folic acid utilizes its method carrying out quantitative detection of folic acid to have commercialization promotional value well.
Further, utilizing the method that the device of quantitative detection of folic acid carries out detection by quantitative to folic acid, linear good in the range of folic acid concentration is 1～18 μM/L especially, average recovery rate is 90%～100%, and RSD% is less than 2%.Therefore the method has the highest sensitivity, precision, accuracy and the response rate, and it is short to detect the time, and service life is long, favorable reproducibility.
Accompanying drawing explanation
Fig. 1, the structural representation of device of a kind of quantitative detection of folic acid of embodiment 1, wherein 1 be aqueous phase platinum filament to electrode II, 2 is aqueous phase reference electrode Ag/AgCl;3 be oil phase platinum filament to electrode I, 4 is oil phase reference electrode Ag/AgCl；5 be mixed solution, 6 be organogel, 7 be MCM-41 type organic/inorganic compound mesoporous film, 8 be the container of upper end open, 9 for hollow pipe；
Fig. 2, the scanning electron microscope (SEM) photograph on MCM-41 type organic/inorganic compound mesoporous film surface of embodiment 1 gained；
Fig. 3, various variable concentrations folic acid standard solution Folic Acid anion be differential pulse Stripping Voltammetry response picture on the device of the quantitative detection of folic acid of embodiment 1；
Fig. 4, YESUAN PIAN sample solution Folic Acid anion be differential pulse Stripping Voltammetry response picture on the device of the quantitative detection of folic acid of embodiment 1；
Curve between Fig. 5, folic acid standard solution peak current folic acid concentration corresponding with standard solution；
Fig. 6, YESUAN PIAN sample solution peak current and the standard curve between folic acid standard solution peak current and the corresponding folic acid concentration of standard solution；
Fig. 7, concentration are folic acid solution Folic Acid anion cyclic voltammetric response diagram on the device of the quantitative detection of folic acid of embodiment 1 of 4 μMs/L；
Fig. 8, concentration are the folic acid solution Folic Acid anion of 4 μMs/L different cyclic voltammetric response diagrams sweeping speed on the device of the quantitative detection of folic acid of embodiment 1；
Fig. 9, concentration are the curve between the folic acid solution Folic Acid anion cyclic voltammetric peak current of 4 μMs/L and the square root sweeping speed；
Figure 10, concentration are folic acid solution Folic Acid anion differential pulse voltammetry response diagram on the device of the quantitative detection of folic acid of embodiment 1 of 4 μMs/L；
Figure 11, concentration are folic acid solution Folic Acid anion differential pulse Stripping Voltammetry response diagram on the device of the quantitative detection of folic acid of embodiment 1 of 4 μMs/L；
Figure 12, concentration are the folic acid solution Folic Acid anion differential pulse Stripping Voltammetry peak current of 4 μMs/L and the curve between the preenrichment time.
Detailed description of the invention
Below by embodiment and combine accompanying drawing the present invention is illustrated further, but it is not limiting as the present invention.
Chemical reagent used by the present invention is analytical pure.
Terephthalate fat (PET) film used, film thickness is about 5 μm, diameter 50mm, and average pore size is 100nm, porosity 1.0%-1.5%, Shanghai Hao Xia nucleopore membranes Science and Technology Ltd..
Measuring instrument used by the present invention is as follows:
Scanning electron microscope, S-3400 type SEM scanning electron microscope, HIT；
Embodiment
1
The device of a kind of quantitative detection of folic acid, including aqueous phase pond and oil phase pond；Oil phase pond is placed in the middle and upper part in aqueous phase pond, and aqueous phase pond can be moved up and down by lowering or hoisting gear；Its structural representation as it is shown in figure 1,
Described aqueous phase pond includes that the container 8 of upper end open, aqueous phase platinum filament are to electrode II 1, aqueous phase reference electrode Ag/AgCl2;
Described oil phase pond includes that organogel electrochemical sensor, oil phase platinum filament are to electrode I 3, oil phase reference electrode Ag/AgCl4；
Described organogel electrochemical sensor is that bottom is stained with MCM-41 type organic/inorganic compound mesoporous film 7 and the hollow pipe 9 of upper opening, bottom-up in hollow pipe 9 is sequentially arranged with organogel 6, mixed solution 5；
The material of described hollow pipe 9 is quartz；
The volume of organogel 6 accounts for the 10% of whole hollow pipe volume, and the volume of mixed solution 5 accounts for the 70% of hollow pipe volume;
Described mixed solution 5 is 10mol L for concentration-1Double (triphenylphosphine) ammonium chloride) aqueous solution and concentration is 0.1mol L-1 NaCl aqueous solution calculate by volume, i.e. 10mol L-1Double (triphenylphosphine) ammonium chloride) aqueous solution: 0.1mol L-1 The ratio that NaCl aqueous solution is 5:6 mix；
Described organogel 6 method as follows is prepared from:
Add the 1 of 5ml, 6-dichloro hexane and 0.059g organic facies supporting electrolyte are in small beaker, 40-60 DEG C of oil bath is heated 10-50min, it is subsequently adding the polrvinyl chloride that 0.5g mean molecule quantity is 22000, it is to slowly warm up to 70-90 DEG C, after the transparent state of solution, controls temperature and be 65-85 DEG C, constant temperature stirring 10-50min, obtains organogel；
Organic facies supporting electrolyte wherein used is prepared by a method comprising the following steps and forms:
Double (triphenylphosphine) ammonium chloride and four (4-chlorphenyl) potassium borate are dissolved in respectively the mixed solution formed for the first alcohol and water of 1:1 by volume, again by two kinds of solution mixing, until there being white precipitate to generate, i.e. prepare organic facies supporting electrolyte, the molar concentration of the most double (triphenylphosphine) ammonium chloride is 0.01mol/L, and the molar concentration of four (4-chlorphenyl) potassium borate is 0.01mol/L.
The preparation of the device of above-mentioned a kind of quantitative detection of folic acid, specifically includes following steps:
(1), the preparation of MCM-41 type organic/inorganic compound mesoporous film 7
nullBy dehydrated alcohol，Positive silicic acid four ester and 2.8mmol/L HCl/water solution are in the ratio mix homogeneously of 1g:1.5g:0.13ml，Put into there-necked flask，It is placed in oil bath,Temperature all controls at 50-80 DEG C，Reaction 90min，Stopping is heated to reflux，The reactant liquor of gained is cooled to room temperature，By cetyl trimethylammonium bromide、The HCl/water solution of dehydrated alcohol and 55mmol/L joins after mixing in the ratio of 1g:10g:2.67ml in the reactant liquor of above-mentioned gained and continues stirring 10-50min，Obtain precursor liquid，Then，Under the conditions of room temperature 25 DEG C，Blank for precursor liquid obtained by 2mL terephthalate adipose membrane is filtered in the way of sucking filtration，Keep sucking filtration state 2min，After the substantially dry of i.e. blank terephthalate adipose membrane surface，Obtain MCM-41 type organic/inorganic compound mesoporous film 7，It is specifically shown in granted patent ZL 2,011 1
Shown in 0098700.5；
(2), the preparation of organogel 6
Add the 1 of 5ml, 6-dichloro hexane and 0.059g organic facies supporting electrolyte are in small beaker, 40-60 DEG C of oil bath is heated 10-50min, being subsequently adding 0.5g mean molecule quantity is 22000 polrvinyl chloride, it is to slowly warm up to 70-90 DEG C, after the transparent state of solution, controls temperature and be 65-85 DEG C, constant temperature stirring 10-50min, obtains organogel 6；
Organic facies supporting electrolyte wherein used is prepared by a method comprising the following steps and forms:
Double (triphenylphosphine) ammonium chloride and four (4-chlorphenyl) potassium borate are dissolved in respectively the mixed solution formed for the first alcohol and water of 1:1 by volume, again by two kinds of solution mixing, until there being white precipitate to generate, i.e. prepare organic facies supporting electrolyte, the molar concentration of the most double (triphenylphosphine) ammonium chloride is 0.01mol/L, and the molar concentration of four (4-chlorphenyl) potassium borate is 0.01mol/L；
(3), the preparation of organogel electrochemical sensor
With silica gel, the MCM-41 type organic/inorganic compound mesoporous film 7 of step (1) gained is bonded at the opening of the channel of the bottom of hollow pipe 9, places at least 12h；
Being injected into bottom with the organogel 6 of glass dropper aspiration step (2) gained again to be stained with in the hollow pipe 9 of MCM-41 type organic/inorganic compound mesoporous film, its addition accounts for the 10% of hollow pipe 9 volume, places at least 1h, in order to make organogel 6 harden;
After the organogel 6 in hollow pipe 9 hardens, being added thereto by mixed solution 5, its addition accounts for the 70% of hollow pipe 9 volume, obtains organogel electrochemical sensor；
Described mixed solution 5 is in 50ml volumetric flask, and adding 5ml concentration is 10mol L-1Double (triphenylphosphine) ammonium chloride) aqueous solution and 6ml concentration is 0.1 mol L-1 NaCl aqueous solution, be settled to 50ml with distilled water after mixing；
(4), the assembling of the device of quantitative detection of folic acid
Fixing device is utilized to be fixed on the container 8 of a upper end open by the described organogel electrochemical sensor of step (3) gained in shielded box, one end with MCM-41 type organic/inorganic compound mesoporous film 7 is downward, and the position with MCM-41 type organic/inorganic compound mesoporous film 7 is positioned at the middle and upper part of container, finally, mixed solution 5 at the hollow pipe 9 of organogel electrochemical sensor inserts oil phase platinum filament to electrode I 3, oil phase reference electrode Ag/AgCl4, aqueous phase platinum filament is inserted to electrode II 1 in the container of upper end open, aqueous phase reference electrode Ag/AgCl2, i.e. complete the assembling of the device of quantitative detection of folic acid；
The set of cells of final composition is as follows:
Pt|Ag|AgCl|10mM BTPPACl+10mM NaCl|10mM
BTPPATPBCl + Gellified 1,6-DCH ‖ xμM FA- + Buffer solution|AgCl|Ag|Pt.
Application Example
1
The device utilizing the quantitative detection of folic acid of embodiment 1 gained carries out the detection of folic acid, specifically includes following steps:
(1), the preparation of folic acid standard solution；
Compound concentration is respectively 1 μM/L, 3 μMs/L, 5 μMs/L, 9 μMs/L, 18 μMs/L, 24 μMs/L, 32 μMs/L, the folic acid standard solution of 40 μMs/L；
The solvent used by preparation of described folic acid standard solution is pH=7.2, containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and the aqueous solution of 0.1mol/L sodium hydroxide, its configuration process will be dissolved in above-mentioned solvent by folic acid, then constant volume is in 50ml volumetric flask, obtain the folic acid standard solution of variable concentrations, be put in darkroom and save backup；
(2), the preparation of folic acid sample solution to be measured；
Testing sample is as a example by YESUAN PIAN, take 1 YESUAN PIAN and put into grinding alms bowl, grind with grinding rod, load 25ml small beaker, 15ml buffer solution 1 ultrasonic dissolution 10min is added in beaker, then 25ml centrifuge tube is proceeded to, closeing the lid, put into centrifuge, revolution is 10000-25000 × g, centrifugal 20-50min, described supernatant proceeds to 50ml volumetric flask buffer solution 2 and is settled to 50ml, then pipettes 5ml with pipet, proceeds to 50ml volumetric flask buffer solution 3 and is settled to 50ml, obtain testing sample solution, put into darkroom and save backup；
Described buffer solution 1, buffer solution 2, buffer solution 3 are pH=7.2, containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and the aqueous solution of 0.1mol/L sodium hydroxide；
(3), utilize the device of quantitative detection of folic acid, measure the peak point current corresponding to folic acid standard solution and folic acid sample solution to be measured by electrochemical workstation；
In the container 8 of the upper end open that folic acid standard solution and folic acid sample solution to be measured are loaded the device of quantitative detection of folic acid, by small lifting table device, the container 8 of upper end open is little by little risen, until folic acid standard solution or folic acid sample solution to be measured by chance touch the MCM-41 type organic/inorganic compound mesoporous film 7 bottom hollow pipe 9, operating process can not make the MCM-41 type organic/inorganic compound mesoporous film 7 bottom hollow pipe 9 be immersed in folic acid standard solution or folic acid sample solution to be measured;
Utilize electrochemical workstation, use the preenrichment function of Differential Pulse Stripping Voltammetry (DPSV), at each technical parameter such as amplitude: 0.01-0.1V, pulse width: 0.01-0.1V, sampling width: 0.01-0.1s, pulse period: 0.1-0.5s, sedimentation potential: 0.1-0.8V, sedimentation time: 60s, sensitivity: 10-6-10-4Under conditions of read peak current as corresponding to testing sample solution in step (1) Folic Acid standard solution and step (2) respectively;
(4), the drafting of standard curve；
With peak current as vertical coordinate, folic acid concentration corresponding to folic acid standard solution is abscissa, draws the curve between the folic acid concentration corresponding to peak current and folic acid standard solution, and matching obtains the standard curve equation of folic acid concentration and peak current；
(5), folic acid sample Folic Acid concentration to be measured
The peak current of folic acid sample to be measured in step (3) is substituted in step (4) the gained folic acid concentration standard curve equation with peak current, and it is calculated the concentration of folic acid sample Folic Acid to be measured, then according to the extension rate of step (2) folic acid to be measured sample, thus the content of folic acid sample Folic Acid to be measured is obtained.
Above-mentioned various variable concentrations folic acid standard solution and YESUAN PIAN sample solution Folic Acid anion displacement behavior on quantitative detection of folic acid device, its result is as shown in Figure 3 and Figure 4.
Differential Pulse Stripping Voltammetry, variable concentrations folic acid standard solution Folic Acid anion is utilized all to go out peak at 0.52V as can be seen from Figure 3, and along with the increase peak current of folic acid concentration increases successively.Differential Pulse Stripping Voltammetry, YESUAN PIAN sample solution Folic Acid anion is utilized to go out peak at 0.52V as can be seen from Figure 4.It is indicated above the organogel electrochemical sensor of the present invention to combine Differential Pulse Stripping Voltammetry, for the detection of folic acid, there is preferable repeatability.
Curve between folic acid concentration corresponding to the peak current drawn and 8 folic acid standard solution is as shown in Figure 5, wherein vertical coordinate is peak current, abscissa is the folic acid concentration corresponding to folic acid standard solution, matching obtains the curvilinear equation of folic acid concentration and peak current as shown in Figure 6, Y=0.02X+0.19 (coefficient R=0.996), the peak current data of folic acid sample solution to be measured are substituted in the curvilinear equation of above-mentioned matching gained, be calculated the folic acid concentration in folic acid sample solution to be measured.Then according to the extension rate of step (2) folic acid to be measured sample, thus the content obtaining YESUAN PIAN sample Folic Acid to be measured is 0.393mg, and its result and the folate content 0.4mg that manufacturer provides are consistent in standard relative deviation allowed band.Thus indicate therefore the method and there is good accuracy.
Application Example
2
The device utilizing the quantitative detection of folic acid of embodiment 1 gained observes folic acid anion displacement behavior on this organogel electrochemical sensor, specifically includes following steps:
(1), folic acid standard solution preparation
With pH=7.2, the folic acid standard solution that solution concentration is 4 μMs/L containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and 0.1mol/L sodium hydroxide, it is put in darkroom and saves backup；
(2) device of quantitative detection of folic acid, is utilized, utilize electrochemical workstation (Shanghai Chen Hua instrument company produces CHI660D type electrochemical workstation) to carry out Electrochemical Detection, utilize cyclic voltammetry (CV), differential pulse voltammetry (DPV) and Differential Pulse Stripping Voltammetry (DPSV) to observe folic acid anion electrochemical response on this organogel electrochemical sensor；
In the container 8 of the upper end open that folic acid standard solution is loaded the device of quantitative detection of folic acid, by small lifting table device, the container 8 of upper end open is little by little risen, until folic acid standard solution or folic acid sample solution to be measured by chance touch the MCM-41 type organic/inorganic compound mesoporous film 7 bottom hollow pipe 9, operating process can not make the MCM-41 type organic/inorganic compound mesoporous film 7 bottom hollow pipe 9 be immersed in folic acid standard solution;
The each technical parameter of CV is high potential: 0.8-1.2V respectively, and electronegative potential: 0-0.2V sweeps speed: 0.005-0.1V/s, scans hop count: 2-10, sensitivity: 10-6-10-4；
The each technical parameter of DPV is initial potential: 0-0.2V respectively, terminates current potential: 0.8-1.2V, pulse width: 0.01-0.1V, sampling width: 0.01-0.1s, pulse period: 0.1-0.5s, sensitivity: 10-6-10-4；
The each technical parameter of DPSV is amplitude: 0.01-0.1V respectively, pulse width: 0.01-0.1V, sampling width: 0.01-0.1s, pulse period: 0.1-0.5s, sedimentation potential: 0.1-0.8V, sedimentation time: 5-100s, sensitivity: 10-6-10-4。
Above-mentioned concentration be the folic acid standard solution Folic Acid anion of 4 μMs/L electrochemical response situation on this organogel electrochemical sensor as shown in Fig. 7, Fig. 8, Fig. 9, Figure 10 and Figure 11:
Utilizing cyclic voltammetry (CV) at blank window as can be seen from Figure 7, the most do not add on the window obtained by folic acid solution, there is orientation transfer in the folic acid in anionic state, goes out peak situation in the position of blank window.Blank window ranges is at 0-1.0V, and it is at 0.55V that folic acid anion goes out peak position；
Peak current increases along with the increase sweeping speed as can be seen from Figure 8；
From fig. 9, it can be seen that peak point current is directly proportional, according to slope of a curve in Fig. 9, by Randles-Sev ik formula: i to the square root sweeping speedp=269An3/2cD1/2v1/2,
Can calculate folic acid anion diffusion coefficient in organogel is 1.2 × 10-6cm2/ s, wherein, A is the effective hole area of composite membrane, and n is the electrically charged number of folic acid, and c is folic acid concentration in aqueous, and D is folic acid diffusion coefficient in organogel, and v is scanning speed.Thus illustrate that folic acid anion is transferred to aqueous phase solution from organogel in the way of linear diffusion controls.
Utilizing differential pulse voltammetry (DPV) folic acid anion to go out peak position as can be seen from Figure 10 is at 0.52V；
From Figure 11 and Figure 12 it can be seen that utilize Stripping Voltammetry difference arteries and veins can draw enrichment time reach 60 seconds after the increase of peak current can tend towards stability.I.e. folic acid anion optimal enrichment time on the organogel electrochemical sensor of the device of quantitative detection of folic acid is 60 seconds.May indicate that the device utilizing quantitative detection of folic acid carries out detection by quantitative and folic acid thereon displacement behavior be can be observed folic acid simultaneously.
Application Example
3
Utilize the device detection folic acid precision test of the quantitative detection of folic acid of embodiment 1 gained, operation repetitive 6 times, specifically include following steps:
(1), the preparation of folic acid sample solution to be measured
Testing sample is as a example by YESUAN PIAN, take 1 YESUAN PIAN and put into grinding alms bowl, grind with grinding rod, load 25ml small beaker, 15ml buffer solution 1 ultrasonic dissolution 10min is added in beaker, then 25ml centrifuge tube is proceeded to, closeing the lid, put into centrifuge, revolution is 10000-25000 × g, centrifugal 20-50min, described supernatant proceeds to 50ml volumetric flask buffer solution 2 and is settled to 50ml, then pipettes 5ml with pipet, proceeds to 50ml volumetric flask buffer solution 3 and is settled to 50ml, obtain testing sample solution, put into darkroom and save backup；
Described buffer solution 1, buffer solution 2, buffer solution 3 are pH=7.2, containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and the aqueous solution of 0.1mol/L sodium hydroxide.
(2), utilize the device of quantitative detection of folic acid, measure the peak point current corresponding to folic acid sample solution to be measured by electrochemical workstation
In the container 8 of the upper end open that folic acid sample solution to be measured is loaded the device of quantitative detection of folic acid, by small lifting table device, the container 8 of upper end open is little by little risen, until folic acid standard solution or folic acid sample solution to be measured by chance touch the MCM-41 type organic/inorganic compound mesoporous film 7 bottom hollow pipe 9, operating process can not make the MCM-41 type organic/inorganic compound mesoporous film 7 bottom hollow pipe 9 be immersed in folic acid sample solution to be measured;
Utilize electrochemical workstation, Differential Pulse Stripping Voltammetry (DPSV) is used to observe the peak point current corresponding to folic acid sample solution Folic Acid anion to be measured, utilize folic acid concentration and the standard curve equation of peak current of step (4) gained of Application Example 1, its result after peak point current is converted into folic acid concentration value will be recorded and see table:
Each technical parameter of described Differential Pulse Stripping Voltammetry (DPSV) is amplitude: 0.01-0.1V respectively, pulse width: 0.01-0.1V, sampling width: 0.01-0.1s, pulse period: 0.1-0.5s, sedimentation potential: 0.1-0.8V, sedimentation time: 60s, sensitivity: 10-6-10-4。
Show from the result of upper table, utilize the device of quantitative detection of folic acid of the present invention method to quantitative detection of folic acid, its detection relative standard deviation is less than 2%, illustrate that the device utilizing the quantitative detection of folic acid of the present invention has the preferable advantage of repeatability to the method for quantitative detection of folic acid for folic acid carries out detection by quantitative, it is possible to meet the requirement of experimental determination folate content.
Application Example
4
Utilize the test to the response rate of quantitative detection of folic acid of the device of quantitative detection of folic acid of embodiment 1 gained, specifically include following steps:
(1) preparation of folic acid sample solution, to be measured
Testing sample is as a example by YESUAN PIAN, take 1 YESUAN PIAN and put into grinding alms bowl, grind with salt frotton, load 25ml small beaker, 15ml buffer solution 1 ultrasonic dissolution 10min is added in beaker, then 25ml centrifuge tube is proceeded to, closeing the lid, put into centrifuge, revolution is 10000-25000 × g, centrifugal 20-50min, described supernatant proceeds to 50ml volumetric flask buffer solution 2 and is settled to 50ml, then pipettes 5ml with pipet, proceeds to 50ml volumetric flask buffer solution 3 and is settled to 50ml, obtain YESUAN PIAN sample solution to be measured, put into darkroom and save backup；
Described buffer solution 1, buffer solution 2, buffer solution 3 are pH=7.2, containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and the aqueous solution of 0.1mol/L sodium hydroxide.
(2), the preparation of response rate sample solution
Take the folic acid sample solution 2 groups to be measured of step (1) gained, its concentration is 1.79 μMs/L, the i.e. background values of response rate sample solution is 1.79 μMs/L, then be added to respectively concentration be 5 μMs/L, 18 μMs/L folic acid standard solution, obtain 2 groups of response rate sample solutions；
(3), utilize the device of quantitative detection of folic acid, measure the peak point current corresponding to 2 groups of response rate sample solutions by electrochemical workstation, often group do 6 parallel
In the container 8 of the upper end open that response rate sample solution is loaded the device of quantitative detection of folic acid, by small lifting table device, the container 8 of upper end open is little by little risen, until folic acid standard solution or folic acid sample solution to be measured by chance touch the MCM-41 type organic/inorganic compound mesoporous film 7 bottom hollow pipe 9, operating process can not make the MCM-41 type organic/inorganic compound mesoporous film 7 bottom hollow pipe 9 be immersed in response rate sample solution;
Utilize electrochemical workstation, Differential Pulse Stripping Voltammetry (DPSV) is used to observe the peak point current situation corresponding to various variable concentrations response rate sample solution Folic Acid aniones, utilize folic acid concentration and the standard curve equation of peak current of step (4) gained of Application Example 1, its result after peak point current is converted into folic acid concentration value will be recorded and see table:
Wherein each technical parameter of DPSV is amplitude: 0.01-0.1V respectively, pulse width: 0.01-0.1V, sampling width: 0.01-0.1s, pulse period: 0.1-0.5s, sedimentation potential: 0.1-0.8V, sedimentation time: 60s, sensitivity: 10-6-10-4。
Result from upper table shows: 2 groups of response rate sample solutions, response rate meansigma methods is all between 90%～100%, RSD be less than 2%, illustrate the device utilizing the quantitative detection of folic acid of the present invention method to quantitative detection of folic acid for folic acid is carried out detection by quantitative, its accuracy is good.
In sum, the device of the quantitative detection of folic acid of the present invention and utilize it to have the highest sensitivity, precision, accuracy, the response rate and repeatability to the method for quantitative detection of folic acid.Further, owing to organogel electrochemical sensor used in the device of quantitative detection of folic acid has, preparation is simple, operate the characteristics such as easy and with low cost.Therefore the device of quantitative detection of folic acid and utilize its method carrying out detection by quantitative medicine Folic Acid to have commercialization promotional value well.
Above said content is only the basic explanation under present inventive concept, and according to any equivalent transformation that technical scheme is done, all should belong to protection scope of the present invention.
Claims (6)
1. the device of a quantitative detection of folic acid, it is characterised in that the device of described quantitative detection of folic acid includes aqueous phase pond and oil phase pond；Oil phase pond is placed in the middle and upper part in aqueous phase pond, and aqueous phase pond can be moved up and down by lowering or hoisting gear；
Described aqueous phase pond includes that the container of upper end open, aqueous phase platinum filament are to electrode II, aqueous phase reference electrode Ag/AgCl;
Described oil phase pond includes that organogel electrochemical sensor, oil phase platinum filament are to electrode I, oil phase reference electrode Ag/AgCl；
Described organogel electrochemical sensor is that bottom is stained with MCM-41 type organic/inorganic compound mesoporous film and bottom-up in the hollow pipe of upper opening, and hollow pipe is sequentially arranged with organogel, mixed solution；
Wherein the volume of organogel accounts for the 10-20% of whole hollow pipe volume, and the volume of mixed solution accounts for the 30-80% of hollow pipe volume；
Described mixed solution be concentration be 10mol L-1Double (triphenylphosphine) aqueous ammonium chloride solutions and concentration be 0.1mol L-1's NaCl aqueous solution calculates by volume, i.e. 10mol L-1Double (triphenylphosphine) aqueous ammonium chloride solutions: 0.1mol L-1The ratio that NaCl aqueous solution is 5:6 mix；
Described organogel method as follows is prepared from:
By 1,6-dichloro hexane and organic facies supporting electrolyte, 10-50min is heated in 40-60 DEG C of oil bath, it is subsequently adding the polrvinyl chloride that mean molecule quantity is 22000, it is to slowly warm up to 70-90 DEG C, after the transparent state of solution, controls temperature and be 65-85 DEG C, constant temperature stirring 10-50min, obtains organogel；
Wherein 1,6-dichloro hexane, organic facies supporting electrolyte and the consumption of polrvinyl chloride that mean molecule quantity is 22000, by 1,6-dichloro hexane: organic facies supporting electrolyte: mean molecule quantity is the ratio that polrvinyl chloride is 5ml:0.059g:0.1-1.0g of 22000；
Wherein said organic facies supporting electrolyte is prepared by a method comprising the following steps and forms:
Double (triphenylphosphine) ammonium chloride and four (4-chlorphenyl) potassium borate are dissolved in respectively the mixed solution formed for the first alcohol and water of 1:1 by volume, again by two kinds of solution mixing, until there being white precipitate to generate, obtain organic facies supporting electrolyte, the molar concentration of the most double (triphenylphosphine) ammonium chloride is 0.01mol/L, and the molar concentration of four (4-chlorphenyl) potassium borate is 0.01mol/L.
The device of a kind of quantitative detection of folic acid the most as claimed in claim 1, it is characterised in that the volume of described organogel accounts for the 10% of whole hollow pipe volume, and the volume of mixed solution accounts for the 70% of hollow pipe volume；
The consumption of described 1,6-dichloro hexane, organic facies supporting electrolyte and polrvinyl chloride that mean molecule quantity is 22000, by 1,6-dichloro hexane: organic facies supporting electrolyte: mean molecule quantity is the ratio that polrvinyl chloride is 5ml:0.059g:0.5g of 22000.
The device of a kind of quantitative detection of folic acid the most as claimed in claim 1 or 2, it is characterised in that the material of the hollow pipe of described upper opening is quartz.
The preparation method of the device of a kind of quantitative detection of folic acid the most as claimed in claim 1, it is characterised in that specifically include following steps:
(1), the preparation of MCM-41 type organic/inorganic compound mesoporous film；
By dehydrated alcohol, the positive own ester of silicic acid four and 2.8mmol/L
HCl/water solution, in the ratio mix homogeneously of 1g:1-5g:0.1-1ml, puts into there-necked flask, is placed in oil bath, and temperature controls at 50-80 DEG C, reacts 90min, stops heating, and the reactant liquor of gained is cooled to room temperature；
Join after the HCl/water solution of cetyl trimethylammonium bromide, dehydrated alcohol and 55mmol/L is mixed in the ratio of 1g:5-15g:1-5ml and the reactant liquor of above-mentioned gained continues stirring 10-50min, obtain precursor liquid, then, under the conditions of room temperature 25 DEG C, blank for precursor liquid obtained by 2mL terephthalate adipose membrane is filtered in the way of sucking filtration, keep sucking filtration state 2min, after the substantially dry of i.e. blank terephthalate adipose membrane surface, obtain MCM-41 type organic/inorganic compound mesoporous film；
(2), the preparation of organogel
By 1,6-dichloro hexane and organic facies supporting electrolyte heat 10-50min in 40-60 DEG C of oil bath, it is subsequently adding the polrvinyl chloride that mean molecule quantity is 22000, it is to slowly warm up to 70-90 DEG C, after the transparent state of solution, control temperature and be 65-85 DEG C, constant temperature stirring 10-50min, obtain organogel；
Wherein 1,6-dichloro hexane, organic facies supporting electrolyte and the consumption of polrvinyl chloride that mean molecule quantity is 22000, by 1,6-dichloro hexane: organic facies supporting electrolyte: mean molecule quantity is the ratio that polrvinyl chloride is 1ml:0.012g:0.02-0.5g of 22000；
(3), the preparation of organogel electrochemical sensor
With silica gel, the MCM-41 type organic/inorganic compound mesoporous film of step (1) gained is bonded at the opening of the channel bottom a hollow pipe, places at least 12h；
Being injected into bottom with glass dropper absorption organogel again to be stained with in the hollow pipe of MCM-41 type organic/inorganic compound mesoporous film, place at least 1h, in order to make organogel harden, the volume of organogel accounts for the 10-20% of whole hollow pipe volume;
After the organogel in hollow pipe hardens, being added thereto by mixed solution, its addition accounts for the 30-80% of hollow pipe volume, obtains organogel electrochemical sensor；
Described mixed solution is in 50ml volumetric flask, and adding 5ml concentration is 10mol L-1Double (triphenylphosphine) aqueous ammonium chloride solutions and 6ml concentration be 0.1mol L-1NaCl aqueous solution after be settled to 50ml with distilled water；
(4), the assembling of the device of quantitative detection of folic acid
Fixing device is utilized to be fixed on the container of a upper end open by organogel electrochemical sensor in shielded box, one end with MCM-41 type organic/inorganic compound mesoporous film is downward, and the position with MCM-41 type organic/inorganic compound mesoporous film is positioned at the middle and upper part of container, finally, oil phase platinum filament is inserted to electrode I in the mixed solution of the hollow pipe of organogel electrochemical sensor, oil phase reference electrode Ag/AgCl, aqueous phase platinum filament is inserted to electrode II in the container of upper end open, aqueous phase pond, aqueous phase reference electrode Ag/AgCl, i.e. complete the assembling of the device of quantitative detection of folic acid.
5. the preparation method of the device of quantitative detection of folic acid as claimed in claim 4, it is characterised in that the blank terephthalate adipose membrane described in step (1), film thickness is 5 μm, diameter 50mm, and average pore size is 100nm, porosity 1.0%-1.5%.
6. the device utilizing quantitative detection of folic acid as claimed in claim 1 or 2 carries out quantitative detecting method to folic acid, it is characterised in that specifically include following steps:
(1), the preparation of folic acid standard solution
Compound concentration is respectively 1 μM/L, 3 μMs/L, 5 μMs/L, 9 μMs/L, 18 μMs/L, 24 μMs/L, 32 μMs/L, the folic acid standard solution of 40 μMs/L；
The solvent used by preparation of described folic acid standard solution is pH=7.2, containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and the aqueous solution of 0.1mol/L sodium hydroxide, its process for preparation will be dissolved in above-mentioned solvent by folic acid, then constant volume is in 50ml volumetric flask, obtain the folic acid standard solution of variable concentrations, be put in darkroom and save backup；
(2) preparation of folic acid sample solution, to be measured
Take 1 YESUAN PIAN and put into grinding alms bowl, grind with grinding rod, load 25ml small beaker, in beaker, add 15ml buffer solution 1 ultrasonic dissolution 10min, then proceed to 25ml centrifuge tube, close the lid, put into centrifuge, revolution is 10000-25000 × g, centrifugal 20-50min, and supernatant proceeds to 50ml volumetric flask buffer solution 2 and is settled to 50ml, 5ml is pipetted again with pipet, proceed to 50ml volumetric flask buffer solution 3 and be settled to 50ml, obtain folic acid sample solution to be measured, put into darkroom and save backup；
Described buffer solution 1, buffer solution 2, buffer solution 3 are pH=7.2, containing 0.1mol/L sodium chloride, 0.1mol/L potassium dihydrogen phosphate and the aqueous solution of 0.1mol/L sodium hydroxide；
(3), utilize the device of quantitative detection of folic acid, measure the peak point current corresponding to folic acid standard solution and folic acid sample solution to be measured by electrochemical workstation；
The container of the upper end open of the device of quantitative detection of folic acid will load folic acid sample solution to be measured or folic acid standard solution, by small lifting table device, the container of upper end open is little by little risen, until folic acid standard solution or folic acid sample solution to be measured by chance touch the MCM-41 type organic/inorganic compound mesoporous film bottom the hollow pipe of organogel electrochemical sensor;
With the preenrichment function of Differential Pulse Stripping Voltammetry, at each technical parameter, amplitude: 0.01-0.1V, pulse width: 0.01-0.1V, sampling width: 0.01-0.1s, pulse period: 0.1-0.5s, sedimentation potential: 0.1-0.8V, sedimentation time: 5-100s, sensitivity: 10-6-10-4Under conditions of the peak current corresponding to folic acid sample solution respectively to be measured in read step (1) Folic Acid standard solution and step (2);
(4), the drafting of standard curve
With peak current as vertical coordinate, folic acid concentration corresponding to folic acid standard solution is abscissa, draws the curve between the folic acid concentration corresponding to peak current and folic acid standard solution, and matching obtains the standard curve equation of folic acid concentration and peak current；
(5), folic acid sample Folic Acid concentration to be measured
The peak current of folic acid sample solution to be measured in step (3) is substituted in step (4) the gained folic acid concentration standard curve equation with peak current, and it is calculated the concentration of folic acid sample solution Folic Acid to be measured, then according to the extension rate of step (2) folic acid to be measured sample, thus the content of folic acid sample Folic Acid to be measured is obtained.
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| CN201410335047.3A CN104198565B (en) | 2014-07-15 | A kind of quantitative detection of folic acid device and preparation method thereof and its application in quantitative detection of folic acid |
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| CN201410335047.3A CN104198565B (en) | 2014-07-15 | A kind of quantitative detection of folic acid device and preparation method thereof and its application in quantitative detection of folic acid |
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