KR102086414B1 - Method for manufacturing nanopipette having an membrane comprising saturated ionophore - Google Patents

Abstract

The present invention discloses a nanopipette having a membrane comprising a saturated ion sensing material, a method of manufacturing the same, and an ion measuring apparatus including the same.

Description

METHODS FOR MANUFACTURING NANOPIPETTE HAVING AN MEMBRANE COMPRISING SATURATED IONOPHORE}

The present invention relates to a method for manufacturing a nanopipette having a membrane comprising a saturated ion sensing material, and more particularly includes an ion selective membrane inside the nanopipette, and the ion selective membrane includes an ion sensing material saturated therein. It relates to a method for producing nanopipettes showing selectivity.

Ion Selective Electrode (ISE) is a kind of membrane electrode having a membrane that selectively responds to a specific ion in a solution. The ion concentration (active amount) can be known by measuring the potential occurring at the interface between the membrane and the solution. . The ion selective electrode is also referred to as a potentiometric ion sensor. The ion selective electrode includes an ion selective membrane, which is the most important part of the ion selective electrode as a membrane that generates voltage by sensing the specific ion in direct contact with the analyte. .

Ion-selective electrodes can measure the concentration of specific ions present in the sample solution, which can be used in clinical studies such as food chemistry, fermentation processes, environmental analysis, hemodialysis, continuous hematoma electrolyte measurement, and extracorporeal blood. It is used in many fields, such as chemical chemistry. On the other hand, the local ion concentration can be measured using a scanning ion-conductance microscope (SICM) with a nanopipette, which can be controlled even at nanoscale units, but the SICM can be measured using the overall ion current ( It is not easy to identify a specific ion distribution because it detects current, or total ion current.

Conventional ISE methods can measure the ions to be detected, but the diameter of the pipette tip is very large in millimeters (mm), and the concentration range of the ions can be measured. there was. However, in order to solve the size problem and enable ion detection even at low concentrations, an ion filter electrode has recently been developed.

The ion filter electrode is a method of making plasticized PVC in the form of a membrane inside the pipette, and can reduce the diameter of the pipette tip to several hundred nanometers. However, the ion filter electrode has a problem that it is not possible to reuse the ion sensing material present in the PVC membrane after use, and to measure one type of ions desired in an aqueous solution containing three or more ions.

In order to solve the problem of the conventional ion filter electrode, the present inventors have applied an ion selective membrane having a saturated ion sensing material inside a nanopipette, so that one or more desired ions can be selectively detected.

According to the first aspect,

An ion-selective nanopipette is disclosed that comprises an ion selective membrane containing an ionophore, a matrix and a plasticizer, the ion sensing material being saturated.

The ion-selective nanopipette according to the first embodiment of the present invention may include an ion selective membrane (middle membrane collector) located centrally in the longitudinal direction of the nanopipette.

The ion-selective nanopipette according to the second embodiment of the present invention may include an ion selective membrane (conical shank membrane collector) located on the conical shank below the nanopipette.

According to the second aspect,

(A) preparing a film forming solution containing an ionophore, a polymer matrix and a plasticizer;

(B) injecting the membrane forming solution into the nanopipette to form an ion selective membrane; And

(C) A method for producing an ion-selective nanopipette comprising the step of saturating an ion sensing material in a membrane.

Method for preparing an ion-selective nanopipette according to the first embodiment of the present invention is:

(a) preparing a film forming solution containing an ion sensing material, a polymer support and a plasticizer;

(b) injecting deionized water into the interior of the nanopipette at the top of the nanopipette;

(c) injecting the membrane formation solution onto the deionized water to form an ion selective membrane; And

(d) injecting an electrolyte onto the ion selective membrane.

The method of manufacturing an ion-selective nanopipette according to the embodiment may further include injecting a solution having the same concentration as the electrolyte injected in step (d) to the lower tip end of the nanopipette.

The ion-selective nanopipette prepared according to the embodiment may comprise an ion selective membrane (middle membrane collector) located centrally in the longitudinal direction of the nanopipette.

Method for producing an ion-selective nanopipette according to a second embodiment of the present invention is:

(a) preparing a film forming solution containing an ion sensing material, a polymer support and a plasticizer;

(b) injecting the membrane forming solution to the tip end of the nanopipette to form an ion selective membrane; And

(c) injecting an electrolyte into the nanopipette from the top of the nanopipette.

The method of manufacturing an ion-selective nanopipette according to the embodiment may further comprise the step of placing the lower tip end of the nanopipette in a solution of the same concentration as the electrolyte injected in step (c).

Ion-selective nanopipettes prepared according to this embodiment may comprise an ion selective membrane (conical shank membrane collector) located on the conical shank below the nanopipette.

According to the third aspect,

Nanopipet comprising a membrane containing an ionophore, a matrix and a plasticizer, an internal electrode in contact with an internal solution, a counter electrode in contact with an external solution, and the internal electrode Disclosed is an ion measuring device including a measuring circuit connecting a counter electrode to a counter electrode, wherein the ion sensing material is saturated. The measurement circuit may include an IV converter (current-volage converter), an amplifier for transferring the voltage difference of the internal electrode relative to the counter electrode, and a detector for detecting the signal.

Nanopipettes comprising membranes containing saturated ion sensing materials according to the present invention can selectively detect desired ions. The nanopipette including the membrane containing the saturated ion sensing material according to the present invention can selectively measure target ions even in a sample solution in which a plurality of ions are mixed. In addition, the nanopipette including the membrane containing the saturated ion sensing material according to the present invention can be reused.

1 shows an example of a cation-ion sensing complex according to the present invention.
Figure 2 illustrates the ion exchange principle of a centrally located ion selective membrane (middle membrane collector) according to the present invention.
3 illustrates the ion exchange principle of a conical shank membrane collector located in a conical shank according to the present invention.
Figure 4 shows a method for producing an ion-selective nanopipette comprising a middle membrane in accordance with the present invention.
FIG. 5 shows a method of making an ion-selective nanopipette comprising a conical shank membrane located in a conical shank according to the present invention.
6 shows an example of an ion selective electrode according to the present invention.
Figure 7 shows the SEM measurement results of the nanopipette prepared according to the present invention.
8 shows the result of measuring ion selectivity of the ion membrane located in the middle according to Experimental Example 1 of the present invention.
Figure 9 shows the result of measuring the ion selectivity of the ion membrane located in the cone shank according to Experimental Example 1 of the present invention.
Figure 10 shows the reproducibility confirmation results of the potassium selective membrane located in the cone-shaped shank according to Experimental Example 2 of the present invention.
Figure 11 shows the reproducibility confirmation results of the sodium selective membrane located in the cone-shaped shank located in Experimental Example 2 of the present invention.

Unless defined otherwise herein, all technical and scientific terms used have the same meaning as commonly understood by one of ordinary skill in the art. Various scientific dictionaries that include the terms included herein are well known and available in the art. Although any methods and materials similar or equivalent to those described herein are found to be used in the practice or testing herein, some methods and materials have been described. Depending on the context used by those of ordinary skill in the art, they may be used in a variety of ways, and therefore should not be construed as limiting the invention to particular methodologies, protocols and reagents.

As used herein, the singular encompasses the plural objects unless the context clearly dictates otherwise. As used herein, unless stated otherwise "or" means "and / or". Moreover, the terms "comprising" as well as other forms, such as "having", "consisting of" and "consisting of" are not limiting.

The numerical range includes the numerical values defined in the range. All maximum numerical limits given throughout this specification include all lower numerical limits as if the lower numerical limits were clearly written. All minimum numerical limitations given throughout this specification include all higher numerical limitations as if the higher numerical limitation were clearly written. All numerical limitations given throughout this specification will include all better numerical ranges within the broader numerical range, as the narrower numerical limitations are clearly written.

The headings provided herein are not to be construed as limiting the following embodiments, as a reference to the specification in various aspects or as a whole.

According to a first aspect, the present invention comprises an ion selective membrane containing an ionophore, a matrix, and a plasticizer, wherein the ion sensing material is saturated. To provide an ion-selective nanopipette.

As used herein, the term "ion" refers to a particular state of an atom or molecule and refers to an atom or molecule that loses or gains electrons. Positively charged ions are called cations and negatively charged ions are called anions. The ions are potassium ions (K ⁺ ), sodium ions (Na ⁺ ), calcium ions (Ca ^{2 +} ), manganese ions (Mn ^{2 +} ), copper ions (Cu ²⁺ ), cerium ions (Ce ^{2 +} ) and hydrogen One or more may be selected from the group consisting of ions (H ⁺ ), but is not limited thereto.

As used herein, the term "nanopipet" refers to a hollow, self-supporting nano-conical tip opening, for example having a tip opening of 0.05 nm to 500 nm, 50 nm to 200 nm, or 100 nm to 150 nm. , Inert, non-biological structure. The hollow structure can be glass or quartz, for example, and can retain fluid therein through the tip opening. The interior of the nanopipette can be selected or modified to minimize nonspecific binding of the analyte. The interior of the nanopipette is generally in the form of an elongated cone having a uniform wall thickness of a single layer of quartz or other biologically inert material, and may be sized to allow insertion of an electrode in contact with the solution in the nanopipette. As used herein, nanopipettes generally have one hole, but nanopipettes with multiple concentric holes can be prepared by pulling the double hole capillary. The outer diameter may generally be less than about 1 μm in the tip region.

As used interchangeably herein, the terms "membrane" or "ion-selective membrane" refer to a membrane that is included in an ion-selective electrode (ISE) to generate a current signal by ion exchange by sensing specific ions directly in contact with an analyte. Meaning, the membrane may generally be composed of an ionophore, a polymer matrix, and a plasticizer.

As used herein, the term "ion sensing material" means a material capable of causing a covalent bond, a coordination reaction or an ion exchange reaction with a specific ion. The ion sensing material is a quaternary ammonium salt (valternary ammonium salt), ballinomycin (valinomycin), ballinomycin derivatives, monensin (monensin), nonactin (nonactin), nonactin derivatives, tertiary amine, metal porphyrin (metal porphyrin), metal phthalocyanine, trifluoroacetophenone, trifluoroacetophenone derivatives, crown ether, dibenzo-18-crown-6 6), one or more may be selected from the group consisting of organophosphorus ion sensing material, organotin ion sensing material, ETH1778, ETH1062, ETH1001, ETH129, ETH149, ETH1644, ETH1117, ETH5214, ETH227 and ETH157, but is not limited thereto. It is not.

An example of a saturated ion-sensing material, ie, a cation-ion sensing complex, according to the present invention is shown in FIG. 1, and the principle of ion exchange of an ion-selective membrane comprising a cation-ion sensing complex according to the present invention is shown in FIG. 2. Indicated.

The term "saturated ion sensing material" as used herein means that the ion sensing material in the membrane combines with a cation to form a "cationic-ion sensing complex".

As shown in FIG. 2, the ion-sensing material which is already saturated by reacting with the ions sensed by the ion-sensing material transmits the corresponding ions in the sample through ion exchange to generate current, thereby enabling selective measurement of ions. That is, in the case of ions such as ions saturated in the ion sensing material in the membrane to form a complex, they pass through the membrane through ion exchange, but otherwise the ion exchange does not occur and reflects. As a result, the same ions as the ions that saturate the ion sensing material pass through the membrane due to ion exchange activity, and signal is not detected when other ions are added.

Accordingly, in the case of a nanopipette having a membrane including a saturated ion sensing material according to the present invention, the type of ions used to saturate the ion sensing material, that is, the type of ions sensed by the ion sensing material are selectively transmitted. The type of ions can be controlled.

As used herein, the term "polymeric support" refers to a structure of a membrane that is properly dispersed and maintained to maintain the properties of the other components of the mixed solution constituting the membrane. The polymer support may be a silicone rubber, a copolymer of poly (bisphenol-A carbonate) and poly (dimethylsiloxane), poly (methylmethacrylate) (PMMA), polyurethane (PUR or PU), polyetherimide ( PEI) and poly (vinyl chloride) (PVC) may be selected from one or more, but is not limited thereto.

As used herein, the term "plasticizer" refers to a nonvolatile organic solvent that is used in high proportions to reduce the rigidity of the polymeric support. The plasticizer is o-nitrophenyl octyl ether, bis (2-ethylhexyl) sebacate, dioctyl phthalate, bis (1-butylpentyl) adipate, dioctyl phenylphosphate, tris (2-ethylhexyl) ester, o- One or more may be selected from the group consisting of nitrophenyl phenylphosphate and dibutyl phthalate, but is not limited thereto.

According to the present invention, the ion selective membrane may further include an anion repeller.

The term "anion repeller" as used herein serves to reflect anions, reflecting the anions to allow the cations to be more easily accessible around the membrane. The anion repeller is triphenylboron, trispentafluoropheneyl boron, tris (3,5-di (trifluoromethyl) phenyl) boron (tris (3,5-di) (trifluoromethyl) phenyl) boron), tetraphenylborate salt, and tetrakis (pentafluorophenyl) borate salt, but may be selected from one or more, but not limited thereto. It doesn't happen.

According to the first embodiment of the present invention, the ion-selective nanopipette may include an ion selective membrane (middle membrane collector) located centrally in the longitudinal direction of the nanopipette.

According to a second embodiment of the present invention, the ion-selective nanopipette may comprise an ion selective membrane (conical shank membrane collector) located on the conical shank below the nanopipette. The ion exchange principle of the ion selective membrane located on the conical shank is shown in FIG. 3.

Specifically, ion exchange of an ion selective membrane located in the lower conical shank of a nanopipette is the same principle as ion exchange of an ion selective membrane centrally located in the longitudinal direction of the nanopipette. However, since the ion-selective membrane located on the conical shank is formed at the tip of the nanopipette and ion exchange occurs at the same time as the sample solution is added, ion exchange activity may occur faster than the ion exchange of the membrane located centrally in the longitudinal direction of the nanopipette. As a result, noise is reduced and a more stable signal can be obtained.

According to a second aspect, the present invention provides a method of making an ion-selective nanopipette. The method comprises the steps of: (A) preparing a film forming solution containing an ionophore, a polymer matrix and a plasticizer; (B) injecting the membrane forming solution into the nanopipette to form an ion selective membrane; And (C) saturating the ion sensing material in the ion selective membrane.

The film forming solution according to the present invention contains an ion sensing material, a polymer support and a plasticizer, and may further contain an anion repeller. Specifically, the film forming solution according to the present invention is bis (benzo-15-crown-5) and / or bis (12-crown-4) as an ion sensing material, PCV as a polymer support, o-nitrophenyl octyl ether as a plasticizer (o-nitrophenyloctylether) and tetraphenylborate as an anionic repeller may be prepared by mixing in a ratio of 5: 32: 62: 1. The film forming solution may be dissolved in tetrahydrofuran. Chemical structures of the ion sensing materials bis (benzo-15-crown-5) and bis (12-crown-4) are shown in Chemical Structure 1 (a) and Chemical Structure 2 (b), respectively.

[Chemical Structure 1]

Referring to the chemical structure, the ring portion of bis (benzo-15-crown-5) and bis (12-crown-4) is negatively charged by the oxygen bond. Thus, potassium ions and sodium ions can bind to the negatively charged portion. Since the size of the potassium ion is about 152 pm and the size of the sodium ion is about 116 pm, it can bind to bis (benzo-15-crown-5) and bis (12-crown-4), respectively.

The ion-selective nanopipette manufacturing method according to the first embodiment of the present invention comprises the steps of (a) preparing a film forming solution containing an ion sensing material, a polymer support and a plasticizer; (b) injecting deionized water into the interior of the nanopipette at the top of the nanopipette; (c) injecting the membrane formation solution onto the deionized water to form an ion selective membrane; And (d) injecting an electrolyte onto the ion selective membrane. The method of manufacturing an ion-selective nanopipette according to the embodiment may further include injecting a solution having the same concentration as the electrolyte injected in step (d) to the lower tip end of the nanopipette. The ion-selective nanopipettes prepared according to the embodiments may comprise a middle membrane located centrally in the longitudinal direction of the nanopipette. 4 shows a method of preparing an ion-selective nanopipette according to the embodiment.

Referring to FIG. 4, first, deionized water is injected from the upper portion of the nanopipette to the lower tip portion of the nanopipette. A membrane forming solution is injected onto the injected deionized water and dried to form an ion selective membrane. After injecting electrolyte into the nanopipette to saturate the ion sensing material in the ion selective membrane, and injecting a solution of the same concentration as the electrolyte at the lower tip of the nanopipette, the ion selective membrane located between the electrolytes Can be saturated. The ions are saturated because the ions (eg, K + and / or Na +) present in the electrolyte combine with the ion sensing material in the ion selective membrane to form a complex. .

According to a second aspect of the present invention, there is provided a method of preparing an ion-selective nanopipette, comprising: (a) preparing a film forming solution containing an ion sensing material, a polymer support, and a plasticizer; (b) injecting the membrane forming solution to the tip end of the nanopipette to form an ion selective membrane; And (c) injecting an electrolyte into the nanopipette from the top of the nanopipette. The method of manufacturing an ion-selective nanopipette according to the embodiment may further comprise the step of placing the lower tip end of the nanopipette in a solution of the same concentration as the electrolyte injected in step (c). Ion-selective nanopipettes prepared according to this embodiment may comprise a membrane (conical shank membrane) located on the conical shank below the nanopipette. 5 shows a method of preparing the ion-selective nanopipette according to the embodiment.

Referring to FIG. 5, first, a membrane-forming solution is injected into a nanopipette and dried to form an ion-selective membrane. After injecting an electrolyte from the top of the nanopipette to the inside of the nanopipette to saturate the ion sensing material in the ion selective membrane, the lower tip end of the nanopipette is placed in a solution of the same concentration as the electrolyte, and both electrolytes Ion-selective membranes located in between can be saturated. The ion selective membrane can be saturated since ions (eg, K + and / or Na +) present in the electrolyte combine with the ion sensing material in the ion selective membrane to form a complex.

According to a third aspect, the present invention provides a nanopipette comprising a membrane containing an ionophore, a polymer matrix, and a plasticizer, an internal electrode in contact with an internal solution, and a counter electrode in contact with an external solution. It includes a (counter electrode) and a measuring circuit for connecting the internal electrode and the counter electrode, the ion sensing material is provided with an ion measuring device characterized in that the saturation. The measurement circuit may include an IV converter (current-volage converter), an amplifier for transferring the voltage difference of the internal electrode relative to the counter electrode, and a detector for detecting the signal. An example of the ion measuring device according to the present invention is shown in FIG. 6.

As used herein, the term "inner solution" means a solution comprising an electrolyte for obtaining an ion current signal. The electrolyte is a material containing free ions, and may be selected from one or more selected from the group consisting of sodium, potassium, calcium, magnesium, chloride, phosphate, and bicarbonate, but is not limited thereto. One or more may be selected, but is not limited thereto.

The term "external solution" as used herein refers to a sample solution containing the ions to be measured, and may include living cells, plasma, and other body fluids.

As used herein, the term “measurement circuit” measures a particular ion signal with alternating current (AC), where the output current follows a change in the input voltage and even a small change in the current can be detected.

The nanopipette including the membrane containing the saturated ion sensing material according to the present invention can selectively detect desired ions according to the kind of ion sensing material and the kind of ions that saturate the ion sensing material. The nanopipette including the membrane containing the saturated ion sensing material according to the present invention can selectively measure target ions in a sample solution having a plurality of ions. In addition, a nanopipette comprising a membrane containing a saturated ion sensing material according to the present invention can reuse the membrane inside the pipette.

Hereinafter, various examples are presented to help understand the invention. The following examples are merely provided to more easily understand the invention, but the protection scope of the invention is not limited to the following examples.

Example

Example  1. Preparation of Nanopipettes

The nano pipette having a diameter of 100 nm was prepared using a puller (Model P-2000, Sutter Instrument) based on a CO ₂ laser using a borosilicate glass capillary tube (GD-1, Narishige) equipped with a filament. In this case, the puller setting used is:

Heat = 350, Filament = 3, Velocity = 30, Delay = 190, Pull = 0; And

Heat = 330, Filament = 2, Velocity = 27, Delay = 180, Pull = 250.

The prepared nanopipette was observed through a scanning electron microscope (SEM), and the results are shown in FIG. 7.

Example  2. Preparation of film forming solution

Ion sensing material [bis (12-crown-4) (Dojindo CAS 80403-59-4), PVC (Aldrich, CAS 9002-86-2), plasticizer [o-nitrophenyloctylether (Aldrich, CAS 3244-41-5)] And an anionic repeller (tetraphenylborate) to prepare a film forming solution comprising a weight ratio of 5: 32: 62: 1. Also, except that bis (benzo-15-crown-5) (Dojindo CAS 69271-98-3)] was used as the ion sensing material, a film forming solution was prepared in the same manner as described above.

Example  3. Preparation of ion collector electrode

(1) Preparation of Middlely Located Ion Selective Membrane

Two nanopipettes prepared in Example 1 were prepared, and DI water was injected from the top of the nanopipette to the lower tip end of the nanopipette using a thin needle. Then, each of the membrane-forming solutions prepared in Example 2 was placed on deionized water injected into the nanopipette to form a membrane by cooling in a dry box for 2 hours. 4 × 10 ⁻³ M of potassium chloride (KCl) and sodium chloride (NaCl) solutions were respectively injected onto the membrane, and the same solution was injected into the lower tip end of the nanopipette. It was then stored in a drybox for 2 days to complete a potassium selective membrane saturated with potassium ions and a sodium selective membrane saturated with sodium ions.

(2) Preparation of Conical Shank Membrane Collector Located on Conical Shank

Two nanopipettes prepared in Example 1 were prepared, and each of the membrane-forming solutions prepared in Example 2 was injected to the ends of the nanopipettes, and then cooled in a dry box for 2 hours to form a membrane. At this time, the nanopipette tip portion was immersed in deionized water to prevent the film forming solution from hardening. Then, a 4 × 10 ⁻³ M potassium chloride (KCl) or sodium chloride (NaCl) solution was injected into the nanopipette from the top of the formed nanopipette, and the tip of the nanopipette was also submerged in the same concentration solution. It was then stored in a drybox for 2 days to complete a potassium selective membrane saturated with potassium ions and a sodium selective membrane saturated with sodium ions.

Experimental Example  1. According to the present invention Saturated  Ion Selectivity Measurement Results of Nanopipettes with Membranes Containing Ion Sensing Materials

(1) Measurement of ion selectivity of the ion membrane located in the middle

A potassium selective membrane saturated with potassium ions prepared in Example 3 (1) by injecting an external solution using the low current ion signal detection system shown in FIG. 6 (a) and by the method shown in FIG. 6 (b). And the signals of potassium and sodium ions on sodium selective membranes saturated with sodium ions. The results are shown in FIG. 8 (a) and 8 (b) are graphs measuring potassium ions and sodium ions, respectively.

As can be seen from FIG. 8 (a), when 0.1M KCl was added to the potassium selective electrode as an external solution, the signal increased, but there was little change in the height of the signal when 0.1M NaCl was added. As can be seen from Figure 8 (b), when the 0.1M NaCl was added to the sodium selective electrode as an external solution, the signal increased, but when the 0.1M KCl was added almost no change in the signal. The ratio of signal to noise (S / N) of the potassium selective membrane and the sodium selective membrane was 1.75 and 1.06, respectively.

(2) Ion Selectivity Measurement of Membranes Located on Conical Shanks

Using the same method as in Experimental Example 1 (1), the signals of potassium and sodium ions were measured for the potassium selective membrane saturated with potassium ions and the sodium selective membrane saturated with sodium ions prepared in Example 3 (2) The results are shown in FIG. 9. 9 (a) and 9 (b) are graphs measuring potassium ions and sodium ions, respectively.

As can be seen from Figure 9 (a), the signal increased when 0.1M KCl was added as an external solution in the potassium selective membrane, but the signal was hardly changed when 0.1M NaCl was added. As can be seen from Figure 9 (b), the signal was increased when 0.1M NaCl was added as an external solution in the sodium selective membrane, but the signal was little changed when 0.1M KCl was added. The ratio of signal to noise (S / N) of the potassium selective membrane and the sodium selective membrane was 21.68 and 16.57, respectively.

From the above, it can be seen that the S / N ratio of the ion selective membrane located in the conical shank is increased by about 15 times or more compared to the ion selective membrane located in the middle. In the middle ion-selective membrane, the electrolyte is present at the tip of the nanopipette. Therefore, it was confirmed that noise was generated by the electrolyte present in the tip portion of the nanopipette before the ions in the external solution reached the membrane.

Experimental Example  2. Conical Shank  Check reproducibility of membrane

Using the same method as Experimental Example 1, the saturation process of the ion-sensitive material in the potassium selective membrane and the ion selective membrane prepared in Example 3 (2) was measured for 2 days, 7 days, and 10 days, respectively, And FIG. 11.

As can be seen from FIG. 10 and FIG. 11, it was confirmed that the target ion can be selectively detected even after 2, 7 and 10 days in the case of the conical shank membrane.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the appended claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

Claims (14)

(A) preparing a film forming solution containing an ionophore, a polymer matrix, a plasticizer and an anion repeller;
(B) injecting the membrane forming solution into a nanopipette to form an ion selective membrane; And
(C) A method for producing an ion-selective nanopipette comprising the step of saturating the ion sensing material in the ion selective membrane,
The ion-selective nanopipette enables selective ion measurement by generating a current signal through ion exchange with the same ions that saturate the ion sensing material in the sample,
The ion is one or more selected from the group consisting of potassium ion (K +), sodium ion (Na +), calcium ion (Ca2 +), manganese ion (Mn2 +), copper ion (Cu2 +), cerium ion (Ce2 +) and hydrogen ion (H +) More than that,
The ion sensing material may be quaternary ammonium salt, valineomycin, monensin, nonactin, tertiary amine, metal porphyrin, metal phthalocyanine (metal) phthalocyanine, trifluoroacetophenone, crown ether, dibenzo-18-crow-6-6, organophosphorus ion sensor and organotin ion sensor One or more selected from the group,
The polymer support may be a silicone rubber, a copolymer of poly (bisphenol-A carbonate) and poly (dimethylsiloxane), poly (methylmethacrylate) (PMMA), polyurethane (PUR or PU), polyetherimide ( One or more selected from the group consisting of PEI) and poly (vinyl chloride) (PVC),
The plasticizer may be selected from the group consisting of bis (2-ethylhexyl) sebacate, dioctyl phthalate, bis (1-butylpentyl) adipate, dioctyl phenylphosphate, tris (2-ethylhexyl) ester, and dibutyl phthalate. Or more than one,
The anion repellers include triphenylboron, trispentafluoropheneylboron, and tris (3,5-di (trifluoromethyl) phenyl) boron (tris (3,5-di). (trifluoromethyl) phenyl) boron) is selected from the group consisting of one or more, the method of producing an ion-selective nanopipette.
The method of claim 1,
In the step (B), the ion-selective nano, comprising injecting deionized water into the inside of the nanopipette from the top of the nanopipette, and injecting the film forming solution prepared in step (A) above the deionized water Method of making pipettes.
The method of claim 2,
Wherein (C), in the step (B) comprises the step of injecting an electrolyte into the interior of the nanopipette on the nanopipette on which the ion-selective membrane is formed, ion-selective nanopipette manufacturing method.
The method of claim 3,
After the step (C), further comprising the step of injecting a solution of the same concentration as the injected electrolyte to the lower tip end of the nanopipette, ion-selective nanopipette manufacturing method.
The method of claim 1,
The step (B) is a method of preparing an ion-selective nanopipette, comprising injecting the film forming solution prepared in step (A) from the top of the nanopipette to the tip end of the nanopipette.
The method of claim 5,
Wherein (C), in the step (B) comprises the step of injecting an electrolyte into the interior of the nanopipette on the nanopipette on which the ion-selective membrane is formed, ion-selective nanopipette manufacturing method.
The method of claim 6,
After the step (C), further comprising the step of placing the lower tip end of the nanopipette in a solution of the same concentration as the injected electrolyte, ion-selective nanopipette manufacturing method.
delete delete delete delete The method according to any one of claims 1 to 7,
The ion sensing material, the polymer support, the plasticizer and the anion repeller is characterized in that the mixture in a weight ratio of 5: 32: 62: 1, the method of producing an ion-selective nanopipette.
delete delete

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