CN110333227A - A kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor and its application in measurement anarexol - Google Patents
A kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor and its application in measurement anarexol Download PDFInfo
- Publication number
- CN110333227A CN110333227A CN201910536350.2A CN201910536350A CN110333227A CN 110333227 A CN110333227 A CN 110333227A CN 201910536350 A CN201910536350 A CN 201910536350A CN 110333227 A CN110333227 A CN 110333227A
- Authority
- CN
- China
- Prior art keywords
- electrode
- anarexol
- graphene
- bipyridyl ruthenium
- naphthols
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Electrochemistry (AREA)
- Plasma & Fusion (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention relates to a kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor and its in the application of measurement anarexol, the present invention is fixed on graphene and bipyridyl ruthenium on gold electrode using Nafion membrane method, detects the electrochemical luminescence of anarexol；The result is that with the photomultiplier tube high pressure of 850V and the sweep speed of 0.1V/s；Use the borate buffer solution of pH=7.5；The concentration of anarexol standard items is 1 × 10‑7~6×10‑ 5Luminous intensity in the range of mol/L meets linear rule；The recovery of standard addition of anarexol is between 99.7% ~ 104.9%.So this method has good sensitivity and stability to inspection anarexol drug, effect is fine.Compared with bare electrode, this method can be improved bipyridyl ruthenium utilization efficiency, reduce environmental pollution, save testing cost, be of great significance to application of the electrogenerated chemiluminescence method in Pharmaceutical Analysis is expanded.
Description
Technical field
The present invention relates to a kind of preparation of Electrochemiluminescsensor sensor and its application, in particular to a kind of naphthols/graphite
Alkene/bipyridyl ruthenium Electrochemiluminescsensor sensor preparation method and its measurement anarexol application.
Background technique
Anarexol English name Cyproheptadine, HCL, two phenyl ring of different name, pyridine in heptan, Cyproheptadinum etc., molecular formula
For C21H22CIN, is white tablets, and auxiliary material is calcium sulfate, starch, dextrin, carboxyrnethyl starch sodium, magnesium stearate；Research has shown that hydrochloric acid
Cyproheptadine is H1Receptor antagonist is a kind of hexahydropyridine class antihistamine, and antihistamine effect is stronger than chlorpheniramine and fenazil,
The allergic reactions such as nettle rash, eczema, anaphylaxis and contact dermatitis, pruitus are currently used primarily in, are anticoagulated bloods, anti-
The essential drugs of allergy class.Root is the latest researches have found that cyproheptadine also treats cushing's disease, acromegalia, the popular parotid gland
A variety of new applications such as inflammation.
Electrochemical luminescence (Electrochemiluminescence, ECL), also referred to as electrogenerated chemiluminescence
(Electrogenerated Chemilumieslceence) be electrochemical reaction and chemiluminescence reaction are be combined with each other it is anti-
It answers, using the optical instruments such as photomultiplier tube measurement luminous intensity and spectrum in electrochemiluminescence analysis, may be implemented to trace
The analysis detection of quantity of material.Electrochemiluminescence analysis not only remains many advantages of chemiluminescence analysis, such as high sensitivity,
The range of linearity is wide, instrument is simple, easily operated and observation facilitates, and also has some traditional chemical luminesceence analyses incomparable
Advantage, such as controlling is strong, selectivity is good and can carry out luminesceence analysis in situ, thus cause the extensive concern of people.Currently,
Be broadly divided into following three kinds of electrogenerated chemiluminescence substances, i.e. bipyridyl ruthenium and its derivative by the type of illuminator, luminol and
Its derivative and quantum dot system.Its principle of Electrochemiluminescprocess process method is illuminator in the electricity for being increased by a certain amount of voltage
Pole surface is electrochemically reacted, and the raw substance of electricity is with each other or other substances of electric raw substance and this system carry out electrification
Reaction is learned, obtains excitation state substance, then return to ground state through excitation state substance and generate optical signal, and detected through photomultiplier tube,
It is recorded after converted electrical number by test macro, and then realizes effective measurement to measurement substance.Currently, electrogenerated chemiluminescence
Analytic approach is concerned in the fields such as immunoassay chemistry, environment life science, medicine, and with the exploitation of new function material
Using and sensor modification technique development, further expanded its application range, technical advantage is significant.On the one hand, sensor
Modification technique improves electrochemiluminescence analysis characteristic, is conducive to improve instrument because the micro-structure on sensor surface can be changed
Selectivity and sensitivity；On the other hand, using naphthols filming performance, will have high-specific surface area, high conductivity and electrocatalysis
Can it is graphene-supported arrive sensor surface, enhance the response effect on modified electrode surface, while by luminescence reagent bipyridyl
Ruthenium is immobilized to arrive sensor surface, is conducive to improve bipyridyl ruthenium utilization efficiency, reduces testing cost and reduces environmental pollution.
High performance liquid chromatography is usually used in the detection of anarexol at present, but this method is not only cumbersome, and
The deficiencies such as the generally existing measurement range of linearity is relatively narrow, detect limit for height, sensitivity is low, cannot achieve to the fast of trace hydrochloric acid cyproheptadine
Speed, accurate, efficiently measurement.Research and utilization anarexol establishes electrogenerated chemiluminescence measurement to bipyridyl ruthenium sensitization
The new method of anarexol is reported according to existing literature it is found that there are no using electrogenerated chemiluminescence method to anarexol
It is detected.
Summary of the invention
The technical problem to be solved by the present invention is providing a kind of naphthols/graphene/bipyridyl ruthenium electrogenerated chemiluminescence sensing
The preparation method of device and its measurement anarexol application, the present invention to examine anarexol drug have well spirit
Sensitivity and stability, effect are fine.
The technical solution for solving above-mentioned technical problem is: a kind of naphthols/graphene/bipyridyl ruthenium electrogenerated chemiluminescence sensing
The preparation method of device, comprising the following steps:
(1) electrode pre-processes: gold electrode is used A1 on deerskin2O3Powder is polished to mirror surface, after being rinsed with water, then it is clear with water ultrasound
It washes, room temperature is dried；Gold electrode, to the luminous pond of electrode and reference electrode threading MPI-E type Electrochemiluminescprocess process system
In, the 100 μ L sulfuric acid that concentration is 0.1mol/L, 200 μ L phosphoric acid buffers of pH=8.5 are added into luminous pond with micropipettor
Liquid is detected, until obtaining stable luminosity curve, pretreatment is finished, and is placed on spare in reagent rack；
(2) production of dressing agent: being (70~80) μ L: (1.4~1.6) by naphthols, isopropanol and secondary distilled water by volume
ML: (5~7) mL meter mixing, it is uniform to be put into supersonic wave cleaning machine ultrasonic mixing, obtains coating solution；Graphene and coating solution are pressed again
The ratio of middle naphthols is that 1.5mg: (70~80) μ L meter mixes graphene with coating solution, is put into supersonic wave cleaning machine ultrasonic mixing
Uniformly；Finally the bipyridyl ruthenium that 1 times of coating solution volume is measured is added thereto, it is equal to put standard ultrasonic cleaning machine ultrasonic mixing into
It is even, dressing agent can be obtained；
(3) modification of working electrode: taking 4~6 μ L dressing agents with micropipettor, drop coating in the gold electrode surfaces pre-processed,
Then it is placed on room temperature in reagent rack to dry, so that it may obtain naphthols/graphene/bipyridyl ruthenium modification gold electrode, i.e. naphthols/stone
Black alkene/bipyridyl ruthenium Electrochemiluminescsensor sensor.
It is of the invention another solution is that above-mentioned naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor exists
Measure the application of anarexol, comprising the following steps:
(1) it is tested using the three-electrode system of electrochemical workstation, Ag/AgCl electrode is as reference electrode, platinum electrode
For to electrode, prepared naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor is working electrode, photomultiplier tube
High pressure is set as 850V, and sweep speed is set as 0.1V/s；
(2) it in the borate buffer of 200 μ L, pH=7.5, is matched by the hydrochloric acid of electrogenerated chemiluminescence system detection various concentration
Pyridine in heptan standard solution records the electrochemiluminescence signal intensity of generation, draws working curve；
(3) anarexol standard solution is replaced to detect anarexol sample solution to be measured.
Nafion is a kind of perfluorosulfonic acid type polymer solution, is easily formed a film, and is used for fixed Ru (bpy) earliest3 2+Molecule structure
Build solid phase electrochemical luminescence sensor.Although Ru (bpy)3 2+Easily in conjunction with Nafion, the material of synthesis is still faced with stability
Low, the problems such as electron transfer rate is slow.So the present invention is fixed Ru (bpy) using graphene/Nafion3 2+To construct electrification
Learn luminescence sensor.Electronegative graphene had both increased the electron transfer rate of material, with Ru (bpy)3 2+Combination again may be used
To inhibit Ru (bpy)3 2+Migrated to the electrically inactive hydrophobic region of Nafion, further improve the stability of sensor with it is sensitive
Degree.
The present invention is fixed on graphene and bipyridyl ruthenium on gold electrode using Nafion membrane method, detects anarexol
Electrochemical luminescence；The result is that with the photomultiplier tube high pressure of 850V and the sweep speed of 0.1V/s；It is slow using the boric acid of pH=7.5
Rush solution；The concentration of anarexol standard items is 1 × 10-7~6×10-5Luminous intensity in the range of mol/L meets line
Property rule；Its equation of linear regression isI (counts) = 287.46×106 C+ 1125.4,R 2It is 0.974；Anarexol
Recovery of standard addition between 99.2% ~ 103.8%.So this method has good sensitivity to inspection anarexol drug
With stability, effect is fine.Compared with bare electrode, this method can be improved bipyridyl ruthenium utilization efficiency, reduce environmental pollution, save
Testing cost is of great significance to application of the electrogenerated chemiluminescence method in Pharmaceutical Analysis is expanded.
The present invention with " bipyridyl ruthenium/nitrogen-doped graphene/Nafion composite modified electrode electrochemical analysis hydrochloric acid is different
The research of promazine " technical solution disclosed in document compares, the difference is that: 1. document analytical technology is electrochemical process,
The present invention is electrogenerated chemiluminescence method, and analysis instrument, method are entirely different；2. the document is using nitrogen-doped graphene, just
It is that nitrogen source is added in graphene after treatment, to change graphene microstructure, is mixed for a kind of modified nitrogen containing nitrogen
Miscellaneous graphene；And graphene of the invention is not by processing；3. on method of modifying the document using first by nitrogen-doped graphene+
The mixed liquor of naphthols is applied to glassy carbon electrode surface, bipyridyl ruthenium is added dropwise again after dry, the effect of bipyridyl ruthenium is not intended to generate
Optical signal is only intended to electro-catalysis test sample chlorpromazine hydrochloride；And the present invention is first to mix graphene, naphthols, bipyridyl ruthenium
It closes uniformly, drop-coated will mixed in gold electrode surfaces, bipyridyl ruthenium is used as electric signal luminescence reagent, and the work of graphene
With for enhancing electrode surface electric conductivity and electro-catalysis bipyridyl ruthenium-anarexol system oxidation-reduction reaction；4. detection point
The sample of analysis is different, and the document is for detecting promethazine hydrochloride, and the present invention is for detecting anarexol.
Detailed description of the invention
Fig. 1 is the luminous intensity of different modifying agent modified electrode.
Fig. 2 is the luminous intensity of different drop coating amount modified electrodes.
Fig. 3 is the luminous intensity comparison diagram of bare electrode and modified electrode.
Fig. 4 is the intensity of light reflection comparison diagram of different pH.
Fig. 5 is the luminous intensity comparison diagram of different scanning rates.
Fig. 6 is the luminous intensity comparison diagram of different voltages.
Fig. 7 is the standard curve of standard anarexol.
Fig. 8 is the stability diagram of anarexol luminous intensity.
Specific embodiment
Embodiment 1: a kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor, including it is following
Step:
(1) electrode pre-processes: gold electrode is used A1 on deerskin2O3Powder is polished to mirror surface, after being rinsed with water, then it is clear with water ultrasound
It washes, room temperature is dried；Gold electrode, to the luminous pond of electrode and reference electrode threading MPI-E type Electrochemiluminescprocess process system
In, the 100 μ L sulfuric acid that concentration is 0.1mol/L, 200 μ L phosphoric acid buffers of pH=8.5 are added into luminous pond with micropipettor
Liquid is detected, until obtaining stable luminosity curve, pretreatment is finished, and is placed on spare in reagent rack；
(2) production of dressing agent: 75 μ L of naphthols (Np), isopropanol 1.5mL, secondary distilled water 6mL are mixed, it is clear to be put into ultrasonic wave
Washing machine ultrasound 3min, obtains coating solution；1.5mg graphene (GP), coating solution are mixed again, are put into supersonic wave cleaning machine ultrasound
3min；Bipyridyl ruthenium (the Ru (bpy) that finally 1 times of coating solution volume is measured 3 2+) be added thereto, obtain dressing agent；Preparing
Dressing agent put standard ultrasonic cleaning machine ultrasound 5min into, make dressing agent be uniformly mixed, dressing agent can be obtained；
(3) modification of working electrode: 5 μ L dressing agents are taken with micropipettor, drop coating is in the gold electrode surfaces pre-processed, then
It is placed on room temperature in reagent rack to dry, so that it may obtain naphthols/graphene/bipyridyl ruthenium modification gold electrode, i.e. GP/Np/Ru
(bpy) 3 2+Electrochemiluminescsensor sensor.
Embodiment 2: a kind of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor as described in Example 1 is being surveyed
Determine the application of anarexol, comprising the following steps:
(1) it is tested using the three-electrode system of electrochemical workstation, Ag/AgCl electrode is as reference electrode, platinum electrode
For to electrode, prepared naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor is working electrode, photomultiplier tube
High pressure is set as 850V, and sweep speed is set as 0.1V/s；
(2) it in the borate buffer of 200 μ L, pH=7.5, is matched by the hydrochloric acid of electrogenerated chemiluminescence system detection various concentration
Pyridine in heptan standard solution records the electrochemiluminescence signal intensity of generation, draws working curve；
(3) anarexol standard solution is replaced to detect anarexol sample solution to be measured.
R&D process of the invention is as follows:
Project one: the luminous intensity test of naked working electrode.
1. instrument parameter is arranged: photomultiplier tube high pressure being set as 850V, sweep speed is set as 0.1V/s, when scanning
Between be set as 17s, Scanning Section is set as 2, and sensitivity is set as 1.e-005 A/V；2. solution is added: buffer solution uses pH value
For 8.0 phosphate buffer solution；Bipyridyl ruthenium concentration is 1 × 10-4mol/L；By working electrode/reference electrode/to electrode assemble
Into in the luminous pond of MPI-E type Electrochemiluminescprocess process system, it is slow that 100 μ L phosphoric acid are added into luminous pond with micropipettor
Fliud flushing, 100 μ L bipyridyl ruthenium solution, 100 μ L sulfuric acid solutions；3. activation system is measured；Measure three groups, every group five times, often
Minor tick 5min and recording detection data.
Project two: the optimization of method of modifying.
1. instrument parameter is arranged: photomultiplier tube high pressure being set as 850V, sweep speed is set as 0.1V/s, when scanning
Between be set as 17s, Scanning Section is set as 2, and sensitivity is set as 1.e-005 A/V；2. adjusting coating solution and bipyridyl ruthenium solution
Proportion；Bipyridyl ruthenium is as luminous agent, and graphene can increase luminous intensity.The volume ratio of two kinds of solution can be lowered such as
It is whole: coating solution/bipyridyl ruthenium solution=1:1；Coating solution/bipyridyl ruthenium solution=1.5:1；Coating solution/bipyridyl ruthenium solution=2.5:
1；Coating solution/bipyridyl ruthenium solution=3:1；3. modifying working electrode, dry at room temperature；4. solution is added: buffer solution uses pH
The phosphate buffer solution that value is 8.0；The concentration of sample anarexol solution is 1 × 10-5mol/L；By working electrode/reference electricity
Pole/electrode is fitted into the luminous pond of MPI-E type Electrochemiluminescprocess process system, with micropipettor to luminous Chi Zhongjia
Enter 200 μ L phosphate buffers, 100 μ L anarexol solution；5. activation system is measured；Three groups of each ratio measuring, often
Group five times, every minor tick 5min simultaneously records experimental data.
Project three: the optimization of drop coating amount.
1. instrument parameter is arranged: photomultiplier tube high pressure being set as 850V, sweep speed is set as 0.1V/s, when scanning
Between be set as 17s, Scanning Section is set as 2, and sensitivity is set as 1.e-005 A/V；2. adjusting drop coating amount: pressing two gained film of project
3 μ L, 4 μ L, 5 μ L, 6 μ L drops are respectively adopted using different drop coating amount modified electrodes with bipyridyl ruthenium optimal proportion in solution
It applies；3. modifying working electrode, dry at room temperature；4. solution is added: buffer solution is molten using the phosphoric acid buffer that pH value is 8.0
Liquid；The concentration of sample anarexol solution is 1 × 10-5mol/L；By working electrode/reference electrode/fit into MPI- to electrode
In the luminous pond of E type Electrochemiluminescprocess process system, 200 μ L phosphate buffers are added into luminous pond with micropipettor,
100 μ L anarexol solution；5. activation system is measured；It is three groups fixed that each drop coating measures, every group five times, every minor tick
5min simultaneously records experimental data.
Project four: the optimization of pH value of buffer solution.
1. instrument parameter is arranged: photomultiplier tube high pressure being set as 850V, sweep speed is set as 0.1V/s, when scanning
Between be set as 17s, Scanning Section is set as 2, and sensitivity is set as 1.e-005 A/V；2. adjust buffer solution pH: by project two,
Three gained optimal conditions of project, adjusts the pH value of buffer solution, phosphate buffer solution pH value is 6.0,6.5,7.0,7.5,8.0,
8.5,9.0, borate buffer solution pH value is 6.0,6.5,7.0,7.5,8.0,8.5,9.0；3. modifying working electrode, at room temperature
It dries；4. solution is added: the concentration of sample anarexol solution is 1 × 10-5mol/L；By working electrode/reference electrode/right
Electrode fits into the luminous pond of MPI-E type Electrochemiluminescprocess process system, is added 200 into luminous pond with micropipettor
μ L buffer, 100 μ L anarexol solution；5. activation system is measured；Each pH value measures one group, every group four times, often
Minor tick 5min simultaneously records experimental data.
Project five: the optimization of sweep speed.
1. instrument parameter is arranged: photomultiplier tube high pressure being set as 850V, sweep time is set as 17s, and Scanning Section is set
It is set to 2, sensitivity is set as 1.e-005 A/V；2. adjusting instrument sweep speed 0.04V/s, 0.06V/s, 0.08V/s, 0.1V/
s,0.12V/s；3. modifying working electrode, coating solution/bipyridyl ruthenium solution=1:1, drop coating amount is 5 μ L, is dried at room temperature；④
Solution is added: the concentration of sample anarexol solution is 1 × 10-5mol/L；Buffer solution is that the borate buffer of pH=7.5 is molten
Liquid；By working electrode/reference electrode/fitted into the luminous pond of MPI-E type Electrochemiluminescprocess process system to electrode, use is micro-
200 μ L buffers, 100 μ L anarexol solution are added into luminous pond for amount pipettor；5. activation system is measured；Often
One group of a determination of scan rate, every group four times, every minor tick 5min simultaneously records experimental data
Project six: the optimization of photomultiplier tube high-voltage value.
1. instrument parameter is arranged: sweep speed is set as 0.1V/s, and sweep time is set as 17s, and Scanning Section is set as 2,
Sensitivity is set as 1.e-005 A/V；2. adjusting photomultiplier tube high-voltage value 650V, 700V, 750V, 800V, 850V, 900V；
3. modifying working electrode, coating solution/bipyridyl ruthenium solution=1:1, drop coating amount is 5 μ L, is dried at room temperature；4. solution is added: sample
The concentration of product anarexol solution is 1 × 10-5mol/L；Buffer solution is the borate buffer solution of pH=7.5；By work electricity
Pole/reference electrode/electrode is fitted into the luminous pond of MPI-E type Electrochemiluminescprocess process system, with micropipettor to
200 μ L buffers, 100 μ L anarexol solution are added in luminous pond；5. activation system is measured；Each photomultiplier transit
Pipe high-voltage value measure one group, every group four times, every minor tick 5min simultaneously records experimental data.
Project seven: the stability of modified electrode.
1. instrument parameter is arranged: photomultiplier tube high pressure being set as 900V, sweep speed is set as 0.1V/s, when scanning
Between be set as 105s, Scanning Section is set as 10, and sensitivity is set as 1.e-005 A/V；2. modifying working electrode, coating solution/connection
Pyridine ruthenium solution=1:1, drop coating amount are 5 μ L, are dried at room temperature；3. solution is added: the concentration of sample anarexol solution is
6×10-6mol/L；Buffer solution is the borate buffer solution of pH=7.5；By working electrode/reference electrode/fitted into electrode
In the luminous pond of MPI-E type Electrochemiluminescprocess process system, 200 μ L buffers are added into luminous pond with micropipettor,
100 μ L anarexol solution；4. activation system is measured；Every 21s measurement is primary, once suspends 5min, METHOD FOR CONTINUOUS DETERMINATION 5
It is secondary and record experimental data.
Project eight: the standard curve of measurement anarexol solution.
1. instrument parameter is arranged: photomultiplier tube high pressure being set as 850V, sweep speed is set as 0.1V/s, when scanning
Between be set as 17s, Scanning Section is set as 2, and sensitivity is set as 1.e-005 A/V；2. adjusting the concentration 1 of anarexol solution
×10-7 、2×10-7、4×10-7、6×10-7、8×10-7、1×10-6、2×10-6、4×10-6、6×10-6、8×10-6、1×
10-5、2×10-5、4×10-5、6×10-5mol/L；It is detected by low concentration to high concentration；3. modifying working electrode, film is molten
Liquid/bipyridyl ruthenium solution=1:1, drop coating amount are 5 μ L, are dried at room temperature；4. solution is added: buffer solution is the boron of pH=7.5
Acid buffering solution；Working electrode/reference electrode/fit into MPI-E type Electrochemiluminescprocess process system to electrode is shone
200 μ L buffers, 100 μ L anarexol solution are added with micropipettor into luminous pond by Chi Zhong；5. activation system, into
Row measurement；One group of each concentration mensuration, every group four times, every minor tick 5min simultaneously records experimental data.
Project nine: sample measurement and mark-on reclaims.
1. instrument parameter is arranged: photomultiplier tube high pressure being set as 850V, sweep speed is set as 0.1V/s, when scanning
Between be set as 17s, Scanning Section is set as 2, and sensitivity is set as 1.e-005 A/V；2. modifying working electrode, coating solution/connection pyrrole
Pyridine ruthenium solution=1:1, drop coating amount are 5 μ L, are dried at room temperature；4. solution is added: buffer solution is that the borate buffer of pH=7.5 is molten
Liquid；By working electrode/reference electrode/fitted into the luminous pond of MPI-E type Electrochemiluminescprocess process system to electrode, use is micro-
200 μ L buffers are added into luminous pond for amount pipettor, and (concentration is 1 × 10 to 50 μ L standard items anarexol solution-5mol/
L), (concentration is 1 × 10 to 50 μ L sample anarexol solution-5mol/L)；5. activation system is measured；Three groups are measured, often
Group five times, every minor tick 5min simultaneously records experimental data.6. according to sample test method, take respectively in the linear range 5 kinds not
Same anarexol spiked levels, and anarexol standard solution is added in cyproheptadin hydrochloride tablet measurement sample, it surveys
Content is calculated with calibration curve method after fixed.
Research and develop result and analysis:
1. the selection of dressing agent.
It compared the luminous intensity of anarexol under each dressing agent, No. 1 dressing agent: coating solution: bipyridyl ruthenium solution=1:
1；No. 2 dressing agents: coating solution: bipyridyl ruthenium solution=1.5:1；No. 3 dressing agents: coating solution: bipyridyl ruthenium solution=2.5:1；No. 4
Dressing agent: coating solution: bipyridyl ruthenium solution=3:1；Analyze result such as Fig. 1.
It is obtained by Fig. 1, anarexol is in No. 1 dressing agent modified electrode, luminous intensity highest, and with bipyridyl ruthenium
The reduction of concentration, luminous intensity are also gradually reducing, it is possible to find out, the concentration of bipyridyl ruthenium has centainly luminous intensity
Influence.Therefore, No. 1 dressing agent modified electrode of experimental selection.
2. the selection of drop coating amount.
It compared the luminous intensity of different drop coating amount modified electrodes；Drop coating amount is respectively 3 μ L, 4 μ L, 5 μ L, 6 μ L, analysis knot
Fruit such as Fig. 2.
It is obtained by Fig. 2, luminous intensity increases with the increase of drop coating amount, and when 5 μ L reaches highest, and luminous intensity is opened later
Begin to reduce, it is possible to find out, drop coating amount not be it is much more advantageous to shining, therefore, the drop coating amount of modified electrode selects 5 μ
L。
3. the comparison of bare electrode and modified electrode.
100 μ L of phosphate buffer of pH=8.0 is added in bare electrode, concentration is 1 × 10-5It is 100 μ L of mol/L anarexol, dense
Degree is 1 × 10-4100 μ L of mol/L bipyridyl ruthenium；200 μ L of phosphate buffer of pH=8.0 is added in modified electrode, concentration is 1 × 10-5100 μ L of mol/L anarexol analyzes result such as Fig. 3.
It can be obtained by Fig. 3, the luminous intensity of bare electrode repairs the luminous intensity of electrode significantly lower than dressing agent, thus demonstrates again that
With No. 1 dressing agent modified electrode to the luminous inspection better effect of anarexol.
4. the selection of buffer solution.
Result such as Fig. 4 is analyzed in the comparison of the luminous intensity of the phosphate buffer and borate buffer of different pH value.
Available from figure 4, luminous value becomes larger to gradually rise before this and gradually decreases again with pH's, and phosphate buffer is in pH value
Luminous value is maximum when 8, and borate buffer luminous value when pH value is 7.5 is maximum；And the Integral luminous value of borate buffer is higher than
The Integral luminous value of phosphate buffer, therefore the borate buffer of buffer solution selection pH=7.5.
5. the selection of instrument sweep speed.
Luminous value of No. 1 dressing agent modified electrode under different scanning rates is compared, result such as Fig. 5 is analyzed.
It can be obtained by Fig. 5, luminous value first increases with increasing for sweep speed, and when sweep speed is 0.1V/s, variation becomes
Gesture slows down, under the premise of guaranteeing good test effect, it is contemplated that and excessively high sweep speed can generate damage to test equipment, so
Sweep speed selects 0.1V/s in follow-up test experiment.
6. the selection of photomultiplier tube high-voltage value.
Luminous value of No. 1 dressing agent modified electrode under different photomultiplier tube high-voltage values is compared, result such as Fig. 6 is analyzed.
It can be obtained by Fig. 6, the luminous intensity of photomultiplier tube high pressure more high electrode is higher.But too high voltage can be to instrument
It causes to damage, so having selected the voltage value of 850V high as the photomultiplier tube of experiment in the case where considering signal-to-noise ratio
Pressure value.
7. sample analysis.
(1) standard curve is established.
Under conditions of impinging upon optimal dressing agent, optimal instrument parameter and optimal buffer solution, anarexol
The luminous intensity of each concentration of standard items on the electrode.Analyze result such as Fig. 7.
It can be obtained by Fig. 7, the concentration of anarexol standard items is 1 × 10-7~6×10-5Strong light in the range of mol/L
Degree meets linear rule；Linear equation isI (counts) = 287.46×106 C+ 1125.4,R 2It is 0.974.So again
Illustrate that the electrode of the dressing agent modification of this experimental study has good effect for measurement anarexol.
(2) the stability of dressing agent.
No. 1 dressing agent modified electrode is had studied to 6 × 10-6The stability of mol/L anarexol measurement, analysis result is such as
Fig. 8.
It can be obtained by Fig. 8, modified electrode is more stable to the strong light of anarexol, so the modified electrode detects salt
Sour cyproheptadine has feasibility.
(3) sample measures.
To anarexol sample 1.00 × 10-5Mol/L concentration is measured mark-on reclaims analysis.It the results are shown in Table 1.
The measurement result of 1 drug anarexol of table
The measurement result of 1 drug anarexol of table
It can be obtained by table 1, the recovery of standard addition of anarexol shows the method for this experiment to salt between 99.2% ~ 103.8%
Sour cyproheptadine drug has good sensitivity, can be good at measurement and analysis anarexol drug.
Claims (2)
1. a kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor, it is characterised in that: including with
Lower step:
(1) electrode pre-processes: gold electrode is used A1 on deerskin2O3Powder is polished to mirror surface, after being rinsed with water, then it is clear with water ultrasound
It washes, room temperature is dried；Gold electrode, to the luminous pond of electrode and reference electrode threading MPI-E type Electrochemiluminescprocess process system
In, the 100 μ L sulfuric acid that concentration is 0.1mol/L, 200 μ L phosphoric acid buffers of pH=8.5 are added into luminous pond with micropipettor
Liquid is detected, until obtaining stable luminosity curve, pretreatment is finished, and is placed on spare in reagent rack；
(2) production of dressing agent: being (70~80) μ L: (1.4~1.6) by naphthols, isopropanol and secondary distilled water by volume
ML: (5~7) mL meter mixing, it is uniform to be put into supersonic wave cleaning machine ultrasonic mixing, obtains coating solution；Graphene and coating solution are pressed again
The ratio of middle naphthols is that 1.5mg: (70~80) μ L meter mixes graphene with coating solution, is put into supersonic wave cleaning machine ultrasonic mixing
Uniformly；Finally the bipyridyl ruthenium that 1 times of coating solution volume is measured is added thereto, it is equal to put standard ultrasonic cleaning machine ultrasonic mixing into
It is even, dressing agent can be obtained；
(3) modification of working electrode: taking 4~6 μ L dressing agents with micropipettor, drop coating in the gold electrode surfaces pre-processed,
Then it is placed on room temperature in reagent rack to dry, so that it may obtain naphthols/graphene/bipyridyl ruthenium modification gold electrode, i.e. naphthols/stone
Black alkene/bipyridyl ruthenium Electrochemiluminescsensor sensor.
2. a kind of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor as described in claim 1 is in measurement hydrochloric acid match
The application of pyridine in heptan, it is characterised in that: the following steps are included:
(1) it is tested using the three-electrode system of electrochemical workstation, Ag/AgCl electrode is as reference electrode, platinum electrode
For to electrode, prepared naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor is working electrode, photomultiplier tube
High pressure is set as 850V, and sweep speed is set as 0.1V/s；
(2) it in the borate buffer of 200 μ L, pH=7.5, is matched by the hydrochloric acid of electrogenerated chemiluminescence system detection various concentration
Pyridine in heptan standard solution records the electrochemiluminescence signal intensity of generation, draws working curve；
(3) anarexol standard solution is replaced to detect anarexol sample solution to be measured.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910536350.2A CN110333227A (en) | 2019-06-20 | 2019-06-20 | A kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor and its application in measurement anarexol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910536350.2A CN110333227A (en) | 2019-06-20 | 2019-06-20 | A kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor and its application in measurement anarexol |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110333227A true CN110333227A (en) | 2019-10-15 |
Family
ID=68142149
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910536350.2A Pending CN110333227A (en) | 2019-06-20 | 2019-06-20 | A kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor and its application in measurement anarexol |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110333227A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111562256A (en) * | 2020-06-11 | 2020-08-21 | 广西科技大学 | Electrochemiluminescence detection method for cyproheptadine hydrochloride |
| CN112858424A (en) * | 2020-12-31 | 2021-05-28 | 广东省现代农业装备研究所 | Preparation method of compound modified electrode for simultaneously detecting lead ions and cadmium ions |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104297229A (en) * | 2014-09-06 | 2015-01-21 | 济南大学 | Making method and application of polythiophene derivative electrochemiluminescence sensor |
| CN105136879A (en) * | 2015-08-20 | 2015-12-09 | 济南大学 | Preparation method and application of electrogenerated chemiluminescence sensor based on graphene quantum dots |
| WO2016015701A1 (en) * | 2014-07-31 | 2016-02-04 | Schebo Biotech Ag | Bioanalysis device, the production thereof and method for detecting bioanalytes by means of the device |
| CN106198503A (en) * | 2016-07-05 | 2016-12-07 | 汕头大学 | A kind of electrochemiluminescence sandwich biosensor and preparation and application |
| CN109001289A (en) * | 2018-07-11 | 2018-12-14 | 广西科技大学鹿山学院 | The method of bipyridyl ruthenium system electrogenerated chemiluminescence method measurement nefopam hydrochloride |
-
2019
- 2019-06-20 CN CN201910536350.2A patent/CN110333227A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016015701A1 (en) * | 2014-07-31 | 2016-02-04 | Schebo Biotech Ag | Bioanalysis device, the production thereof and method for detecting bioanalytes by means of the device |
| CN104297229A (en) * | 2014-09-06 | 2015-01-21 | 济南大学 | Making method and application of polythiophene derivative electrochemiluminescence sensor |
| CN105136879A (en) * | 2015-08-20 | 2015-12-09 | 济南大学 | Preparation method and application of electrogenerated chemiluminescence sensor based on graphene quantum dots |
| CN106198503A (en) * | 2016-07-05 | 2016-12-07 | 汕头大学 | A kind of electrochemiluminescence sandwich biosensor and preparation and application |
| CN109001289A (en) * | 2018-07-11 | 2018-12-14 | 广西科技大学鹿山学院 | The method of bipyridyl ruthenium system electrogenerated chemiluminescence method measurement nefopam hydrochloride |
Non-Patent Citations (2)
| Title |
|---|
| 熊海涛等: "石墨烯修饰电极电化学发光分析法测定甲巯咪唑", 《分析测试学报》 * |
| 贾雪平: "氧化石墨烯的光电化学性质的研究及其应用", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111562256A (en) * | 2020-06-11 | 2020-08-21 | 广西科技大学 | Electrochemiluminescence detection method for cyproheptadine hydrochloride |
| CN111562256B (en) * | 2020-06-11 | 2023-12-26 | 西湖维泰(杭州)诊断技术有限公司 | Electrochemiluminescence detection method for cyproheptadine hydrochloride |
| CN112858424A (en) * | 2020-12-31 | 2021-05-28 | 广东省现代农业装备研究所 | Preparation method of compound modified electrode for simultaneously detecting lead ions and cadmium ions |
| CN112858424B (en) * | 2020-12-31 | 2023-05-23 | 广东省现代农业装备研究所 | Preparation method of compound modified electrode for simultaneously detecting lead and cadmium ions |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Justino et al. | Review of analytical figures of merit of sensors and biosensors in clinical applications | |
| Wang et al. | Sensitive immunoassay of a biomarker tumor necrosis factor-α based on poly (guanine)-functionalized silica nanoparticle label | |
| Wilson et al. | Multiplex measurement of seven tumor markers using an electrochemical protein chip | |
| CN102072931B (en) | Method for preparing biosensor based on silicon nanowires and application of biosensor in detecting DNA | |
| Chen et al. | Detection of total and A1c-glycosylated hemoglobin in human whole blood using sandwich immunoassays on polydimethylsiloxane-based antibody microarrays | |
| Wei et al. | Electrochemical assay of the alpha fetoprotein-L3 isoform ratio to improve the diagnostic accuracy of hepatocellular carcinoma | |
| CN1865959A (en) | Electrode array microchip sensor, preparation method and application therefor | |
| Zhang et al. | Rapid diagnosis of multidrug resistance in cancer by electrochemical sensor based on carbon nanotubes–drug supramolecular nanocomposites | |
| Yicong et al. | A novel microphysiometer based on MLAPS for drugs screening | |
| CN110333227A (en) | A kind of preparation method of naphthols/graphene/bipyridyl ruthenium Electrochemiluminescsensor sensor and its application in measurement anarexol | |
| Stefan et al. | Monocrystalline diamond paste-based electrodes and their applications for the determination of Fe (II) in vitamins | |
| Wang et al. | Immunophenotyping of acute leukemia using an integrated piezoelectric immunosensor array | |
| Shi et al. | A one-piece lateral flow impedimetric test strip for label-free clenbuterol detection | |
| CN102980935A (en) | Electrochemical method for detecting anthracene-phenanthrene resultant of polycyclic aromatic hydrocarbon | |
| Wang et al. | Electrochemiluminescence immunosensor based on the quenching effect of CuO@ GO on m-CNNS for cTnI detection | |
| Li et al. | Clinically applicable homogeneous assay for serological diagnosis of alpha-fetoprotein by impact electrochemistry | |
| Zhang et al. | A facile integrated microfluidic chip based on Chitosan-Gold Nanoparticles-Anchored Three-Dimensional graphene fiber film for monitoring prostate specific antigen | |
| CN103884748B (en) | A kind of preparation method of the molecular imprinting electrochemical sensor for detecting serotonin | |
| Chen et al. | Boronic acid-containing carbon dots array for sensitive identification of glycoproteins and cancer cells | |
| Chanarsa et al. | A sandwich-like configuration with a signal amplification strategy using a methylene blue/aptamer complex on a heterojunction 2D MoSe2/2D WSe2 electrode: Toward a portable and sensitive electrochemical alpha-fetoprotein immunoassay | |
| CN104614421B (en) | A kind of electrochemical method for detecting 2,4,6 trichlorophenol, 2,4,6,-Ts | |
| Li et al. | Disposable Sandwich‐type Electrochemical Sensor for Selective Detection of Glucose Based on Boronate Affinity | |
| Han et al. | Dynamic mapping of electrochemiluminescence reactivity in space: application to bead-based assays | |
| Liang et al. | Flow-injection immuno-bioassay for interleukin-6 in humans based on gold nanoparticles modified screen-printed graphite electrodes | |
| Lee et al. | Electrical detection-based analytic biodevice technology |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20191015 |
|
| WD01 | Invention patent application deemed withdrawn after publication |