DE2638993A1 - ELECTROMOLECULAR MOTION IN VARIOUS SEMI-CONDUCTING MEDIA - Google Patents

Description

Dr.-lng. E. BERKENFELD · Dipl.-Ing. H. BERKENFELD, patent attorneys, Cologne

Απ.ο _ββ file number H 12 ^ / 1

for entering the _{Nome d} . _Note

NORMAN HABER,

12 Churchill Drive,

Old Tappan, New Jersey,

UNITED STATES.

Electromolecular motion in various semiconducting

media

The invention relates to a method for stimulating a chemical species to achieve motility for Orientation, storage and transport, as well as the separation of species, achieved by treating in the appropriate conductivity range of the media and especially within the semiconducting area, induced with the help of intense electrical Fields at or near minimum and optimal current values. Such systems are extremely rapid Molecular movement or transport, subsequently electromolecular Movement (EMB), as well as through great differentiation or dissolution of the molecular types or species.

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.'JoIcIk; Resolution makes it possible to achieve very refined analytical separations.

Compared with traditional techniques, birslancj can be unattainable or unique mobility and versatility of the system can be achieved. The invention creates a Method of inducing mobility in molecules that had previously been considered immobile due to their non-polar nature. In the case of polar molecules, e.g. certain metal derivatives, a greater resolution is obtained than with conventional conductive or aqueous electrolytes .

One aspect of the dBr invention relates to the manufacture of suitable ones Media and systems within which the semiconducting molecule transport can be achieved reliably.

The conductivity of the entire system or process is shifted into the semiconducting range by increasing the level of conductivity the media representing the mobile phase is discontinued. Very high voltages can occur with low currents be maintained so that the thermoelectric heat dissipation

standing (I RT) nevertheless the use of readily available materials and techniques for work systems allowed. In contrast According to the invention, very low currents are actually required for conventional electrochemical transport methods, which correspond to the semiconducting nature of the process. This often excludes the need to use facilities for external heat convection and enables small work arrangements and low requirements on the dimensions the power supply. Another advantage of the method is that with the low heat values according to the invention Diffusion influence is brought to a minimum. The very low current values that are sufficient according to the invention are nearly optimal for molecular movement as they move through

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the attractive-repulsive interaction is induced within the electric field, and among such dedicators a very intense migration effect can be induced which is proportional to the applied voltage potential. This migration effect is characteristic of the molecular nature of the material and can range from such similar or related, though not identical _v structured molecules are sharply differentiated. The characteristic mobility or mobility of a substance in cm / sec can be used to classify or identify substances. The high degree of molecular resolution or differentiation can be achieved over a distance of a few inches in seconds or minutes, requiring relatively less time over small distances - or by using higher voltages. It has been found that certain low current values are almost optimal for the EMB process, and they are defined here as a threshold value function as a function of the molecular type of the materials involved. The threshold value relates to the excited states in a solvation adsorption system. The usual observed areas

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before are 2 10 to 1.6 χ 10 A / cm for a cellulosic substrate. Such threshold values relate to the minimum power requirement to start the EMB process and are usually located close to the optimal power requirement for a given system. The semiconducting domain relates to methods of achieving a suitable conductivity at high voltage at the threshold range.

The method according to the invention can be carried out as a semiconductor transport in the liquid state or in the gas state.

It is a semiconductor fluid process because of its ability to perform molecular shifts and the use of a mobile phase. This distinguishes it from the stationary solid and amorphous semiconductor systems. Because of

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its influence on the electromolecular nature of materials via induction by and reaction with suitable electrical Fields, this method applies to the major classes of known molecular substances, including inorganic ions, organic molecules, colloids and crystalloids. Therefore, this procedure is applicable to inorganic materials, such as those from iron, copper, nickel, cobalt, rare earths, heavy metals, zirconium and the separation of ionic solvates from metal derivatives. It is also applicable to other materials such as proteins, antibiotics, vitamins, antihistamines, amino acids, Dyes and blood components.

In preparative chemistry, under suitable EMB conditions chemical reactions carried out to shift reaction equilibria in favor of certain yields can be applied. There is a possibility of selective consumption of the equilibrium product or of impurities or by-products in the reaction zone. Acts in the extraction the EMB procedure as a minimal time consuming one Procedure.

In applied chemistry, it is applicable where processing or processing requires very rapid and / or selective penetration is desirable, e.g. when dyeing or bleaching fabrics. The dyes or other detectable molecules in a mixture can be customized in a preselected or ordered pattern by controlling its EMB behavior individually to be deposited.

Another Advantage.der invention is that they Separation, identification or study of molecular species due to the different threshold values. It enables control at different values among different ones Conditions of pH, temperature, various media, or other internal or external factors. An

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Application for this would be a procedure that is first carried out by operation of the system with a lower threshold value for the first separation and then through transition to subsequent words is controllable to complete the dissolution.

Main features for operation in the practice of the invention are:

1. Set the working or operating phase to the semiconducting range in order to operate at or near the molecular thresholds and maximum or reasonable stress levels maintained by the system can, enable.

2. Establishing the optimal current value at or near the molecular threshold value at the given voltage for effective molecular dissolution.

3. Applying those components within the system and adjusting the system characteristics so that overall Stability, reproducibility and safety can be achieved.

A useful analogy of this phenomenon and its relationship to electrochemistry is to compare the semiconductor physics of the solid state with the earlier thermionic electrical engineering. Some similarities are the following characteristics of the EMB process refer to:

1. The requirements for consumption (and dimensions) of the power supply are reduced to a minimum.

2. Minimal electrothermal losses allow low Working dimensions and increased field intensities; this contributes to fast resolution times with low distortion values at.

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3. The deteriorating influence on the system as a consequence the need for high currents and the associated thermal effects is eliminated. For example At higher current densities than those used according to the invention, the mobility and resolution of the Molecular type can be changed.

4. The extent and manner of electrical use is not limited to the more conventional conductivity modes such as ion transport in the liquid phase in aqueous electrolyte. Hence, numerous different types of materials can be studied or, used in the EMB process. In addition

• listen to materials and systems, their electrical or ionic contribution would initially be considered poor in terms of their molecular structures. aside from that a large number of non-aqueous, hydrophobic and otherwise non-polar substances as well as ionic, polar, covalent, aprotic or other conductive substances.

5. Operation at or near the lower thresholds can be achieved with an overall high electrical movement output. These threshold values are characteristic of a material and are generally included very low current values.

The method is able to work at current values that are currently sufficient to cause the movement of the molecular species / whereby the electrothermal losses reach negligible values.

Counteracting fractions include evaporative cooling, the. Heat capacity of the container, the thermal convection and the electroendosmotic flow. The controlled loading

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When the threshold values rise, the various types of molecules, in turn, become active in the case of the appropriate and characteristic values are stimulated. This leads to an additional high resolution technique that leads to a differential Molecular discrimination is capable. This method of differentiation is further determined by the speed of movement improved, which is also characteristic of the type of molecule involved. This speed can be changed by modifying the Media can be varied considerably.

Regarding the mechanism of the motion threshold, it is found that this behavior determines the point at which the entire energy supplied counteracts the molecular attraction or adhesion to the .Substrat (surface). It consists of the electrical energy supplied from the outside plus that which is otherwise based on distribution based on additional distribution functions. The molecules can then move freely or be moved by electrical attraction or other convective factors. The electrical characteristic of the system shows a non-linearity as the current gradually increases after the initial application of a given voltage. The preferred systems stabilize rapidly and remain electrically balanced during the separation process, although the process can be carried out when there are gradual changes in electrical properties. In cases where a lack of stability leads to difficulties, but the medium is otherwise considered useful, "the rate of change in the resistance of the system can be reduced by adding a sufficiently large external resistance, for example about the same or greater than that Internal resistance of the system. Alternatively, an active electrical element can be used that can sense the current, voltage or temperature values within a system and is used to regulate these factors or changes with the help of a control of the power source. This practice is also valuable as a safety measure.

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Investigations of components for the composition of the media have shown that certain combinations of compounds for Use with EMB are unsuitable if stable current values are desired. If a constant voltage is applied, show these combinations constantly have a different resistance, as if a capacitor were being charged. This effect can can be illustrated in the form of a current / time diagram at constant voltage, where t. and t ~ on the order of several seconds or minutes:

current

t _x t ₂ time

A material or mixture with electrical properties of the Type A (steadily increasing current) can possibly experience arcing. A material or mixture of type C (ascending Current, then decreasing current after a point) evaporative heating and burns, charred or dries out. A material or mixture of type B is under a preferred medium from the point of view of electrical control, since it enables reproducibility in operation and the Restoring the electrical properties is not a problem. Materials with an electrical behavior of type A. can be freely selected as components for the medium in order to compensate for properties of a medium that would otherwise affect its behavior of type C and vice versa. The electrical properties of a given connection as a function from the other substances with which it is mixed. Examples of connections that affect the behavior of the Illustrating Type A in mixtures and Type B in other mixtures are given below. Generally will to choose a component for a medium · which behaves in type B in conjunction with the other components

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the system shows.

Connections behavior type

N, N-dimethylacetamide in water A

N, N-dimethylacetamide in formamide A

N, N-dimethylformamide in water B

cycl. 1,2-propanediol carbonate in water A

Ethylene carbonate in water B

3-methylsulfolane in water A.

2-pyrrolidinone in water A

N-methylformamide in water A.

N-methylacetamide in water B

Tetrahydrofurfuryl alcohol in water B

Tetrahydrothiophene dioxide in formamide A

Diacetone alcohol in formamide A

Diacetone alcohol in thiodiethylene glycol B

Cellosolve in formamide A

Cellosolve in thiodiethylene glycol B

The EMB procedure differs from the known procedures electrophoresis and dielectrophoresis in various ways.

The EMB response does not seem to be due to the viscosity to be very adversely affected; it is improved by increasing the dielectric constant in the solution, while the electrophoresis mobility is inversely proportional to the dielectric constant, the EMB be carried out at dielectric values that go far beyond the values that are practicable for electrophoresis are. The EMB was carried out in media with a dielectric constant of up to 190, e.g. in N-methylacetamide .

The migration speed in cm / sec of transported by EMB chemical particles is well above the speeds achieved with dielectrophoresis and electrophoresis. The-

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This difference is illustrated by the following diagram:.

log the migration speed
age

\ Electrop

horese

Dielectrophoresis

semiconducting area conductivity

Conduction of the electrolyte

The EMB process is presumably based on proton donor / acceptor interactions and the formation of electron charge transfer (charge transfer) complexes for transport. See. R. Foster, Organic Charge-Transfer Complexes (1969).

In practical terms, a major aspect of this method involves the use of a relatively non-conductive medium. A wide variety of media and techniques can be used to meet the requirements of the semiconducting domains employed. The conduction can be carried out in solids, in semi-solid substances such as gels, as well as in the gas phase, in aerosols, foams and liquids. Combinations of these are also practical, such as melting, high-temperature melting, pseudo-crystals (paracrystals and mesomorphic substances), ice, sludge, smear or suspensions, glasses, plastics, fibers, threads, porous materials and powders. Ion exchange materials, selectively permeable and membrane-like barriers, dialysis membranes, molecular sieves and special ion source materials are suitable as carriers or barriers. The process can be carried out continuously or batchwise. .

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Many substances are relatively dielectric; Among these, the non-polar organic substances represent a large group. Some of these show intermediate ranges of conductivity or are accessible to a suitable adjustment of their conductive nature by the addition of relatively small amounts of additives. This may be similar to the method of doping or of introducing or inoculating as is used in solid-phase arrangements. A relatively polar material can be used as the medium, such as aqueous solutions, by limiting the ion content of the system in order to achieve the desired conductivity value. Also, suppressive substances can be added to the conductive system, among which desirable materials are those which have a suppressive effect in addition to the pure dilution effect that their presence contributes to the system. Furthermore, the suppressive effect of non-polar materials used in admixture with otherwise conductive systems offers a very general and useful solution for controlling conductivity. It is important to note that in relation to all of these techniques, other factors may favor certain additional properties and characteristics of the materials used, suitable for the nature of the application, such as miscibility, compatibility, toxicity, boiling point, melting point, reactivity, cost, Removability, dialysability and osmolality. Often, a high dielectric constant material is preferred because of its ability to hold charges (involving solvation or interaction ) created in the system or charges otherwise acquired or induced on chemical particles.

Mixtures of solvents can be used with the intermediary of a coupling agent, usually of a semi-polar cosolvent nature will. The term semipolar is used for a material that shows some conductivity that after dilution with Water (or other similarly polar material) increases

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and which increases after the addition of a soluble ionic salt. Thus, within the scope of the invention, the solvation of a strongly ionic material in a non-polar material leads through means of a semi-polar material generally leads to only a slight increase in conductivity, while the solution of the ionic material in the semipolar solvent alone can be moderately conductive. Indeed, it can be non-polar Material can be viewed as suppressive of the skills for moderate guidance towards the formation of a three-component system. The three component system therefore comprises an inert base, a conductivity agent and a semi-polar material such as e.g. Xylene, ammonium bromide or dimethylformamide. Furthermore, a significant increase in the amount of the semi-polar solvent can be used improve the conductivity only minimally. The addition of a proportionate small volume of a second type of semi-polar solvent (a four-component system) can then be a cause a considerable increase in conductivity of the entire system. No cosolvent alone with the non-polar material, with or without the solvated ionic material, will achieve the conductivity value thus achieved. This technique for Increasing the conductivity of essentially non-polar materials forms a convenient working basis for using Substances such as xylene, p-cymene, mineral oil and chlorinated solvents. A four component system is for illustration e.g. the system xylene, ammonium bromide, dimethylacetamide and dimethylformamide.

The above effects can also be applied to systems that are not easy Are ionizable, and the components applied by factors such as dielectric constant and proton donor capacity the solvating molecules can be determined. While medium donor capacities give rise to solvated molecules can, can have a high donor capacity in a high dielectric System to easily retain the ionic charges thus generated. Media with a

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Dielectric constants above 10, which tend to increase charges formed by proton donor-acceptor exchange keep.

The media used according to the invention are characterized in that they are liquid at or near room temperature and have a sufficiently high boiling point to withstand the process heat. The boiling points are generally above 140 ° C., preferably above 165 ^° C. The media do not have to be able to dissolve the chemical substance to which the mobility is to be imparted, but the solubility is preferable for the separation of different types of molecules.

The invention is further illustrated by the following examples, which are aimed at the separation of chemical substances in the specified media. The device consisted of a separating cell made of polyethylene divided into two 15 ml chambers high density.

The cell was built to withstand the high voltage fields and against these and against a conductive leak or leakage of media under such fields and kept the many strong and corrosive solvents that are used here safe from leaking. One The platinum electrode in each chamber was connected to a direct current source which generally operated at 1.25 mA. the The power source could be used in certain work areas at different voltage values in the ranges from 0 to 100 μΑ, 0 to 1 mA and 0 to 10 mA for threshold tests and the procedures described here are carried out. A wick made of filter paper in each compartment was with the opposite ends of the filter paper substrate protruding from the top of the cell. The filter paper had a width of 5 cm and a length of 10 cm and, unless otherwise stated, was Vlhatman No. 3. Usually occurs the voltage drop in this system is essentially across

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the impregnated carrier, e.g., from 70 to 90% or more. The cell was through a transparent cover locked in.

A suitable solvent can be selected from the class of low molecular weight Glycols with a small amount of a conductivity-increasing additive can be selected. The following solvent systems are for relatively non-polar dyes as well as for other soluble organic materials useful. The solvents listed were used to separate chemical mixtures, such as dyes, mercurochrome and sodium riboflavin phosphate at the stated values the voltage and current used. The term "stabilized" is used to mean that the electrical Parameters reached the specified values and for the remained constant for a few minutes (generally 2 to 10 minutes), in which the separation process was completed.

Examples of compositions of the solvent electrical game · parameter

. - (stabilized)

1 0.3 ml water, 0.2 ml Sörensen buffer (pH 11 kV / mA 7, O), 24.5 ml of eropylene glycol. The amount of water in these type of systems should preferably not to exceed about 2%.

2 2.0 ml dimethylacetamide, 1.0 ml phenol, 8 kV / mA. 25.0 ml propylene glycol. .

3 2.5 ml formamide, 22.5 ml propylene glycol. 5.5 kV / mA.

Example 3 is excellently suited for the dye dissolution of the following mixture: saframine 0, toluene red (neutral red) and sodium riboflavin phosphate. This medium is also useful for separating members of the rhodamine dye family.

This is different from conventional ion transport processes

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Mobilization of metal derivatives is not easily achieved even if the metal derivatives are soluble in the medium. However, by adjusting the medium and electrical characteristics in accordance with the invention, very fine resolution is achieved in a system such as the following which illustrates a new mode of operation as described herein. Examples of suitable metal ions are Co
eg the chlorides and nitrates.

suitable metal ions are Co, Cu, Ni from salts, such as

Example composition of the solvent electrical

Parameter (stabilized)

10 ml dimethylformamide, 15 ml prop- 8 kV / mA ylene glycol and (if necessary) 1 ml Triethanolamine.

A non-aqueous ester-based system is illustrated below and also leads to satisfactory results. Instead of the cellosolve in the medium, other, related Compounds are used, such as hexyl-cellosolve, Methyl carbitol, cellosolve acetate and carbitol acetate.

Example composition of the solvent electrical

Parameter . \ (stabilized)

5 4 ml formamide, 14 ml cellosolve, 15 kV / mA 34 ml dimethyl phthalate.

The following solvent systems are useful for the separation of metal ions and complexes; of these metal complexes , dithizones, nitroso-B-napthol, pyrocatechol-violet, rhodamine B, 8-hydroxyquinoline and dibenzoylmethane derivatives were used.

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Example composition of the solvent

30 ml methoxy-ethoxy-ethanol + 30 ml cyclic 1,2-propanediol carbonate + 3 drops of nitric acid (1:30 in

H ₂ O)

electrical

Parameter

(stabilized)

6.4 kV / 1.25 mA

The medium of example 6 gave a multi-zone resolution (5 min) with 8-hydroxyquinolinates of rare earths such as Sc and Eu, as well as other metals such as Ni. The heavy metal derivatives of dibenzoylmethane and rhodamine showed good to excellent Movement, while movement was very sparse with heavy metal nitrates and not at all with hafnium (as chloride)

entered. Satisfactory mobility was also obtained for Co Ni ⁺² (as chlorides).

+2

Cu ⁺² and

In the previous example, the nickel chloride resulted in 3 zones with staining of blue and purple. Such reproducible effects show the very high resolution of the technology. This also suggests towards the formation of a number of metal complexes, such as through proton donor-acceptor exchange, and the ability the technique of differentiating and resolving these. This unusual ability occurs in another situation Days where not only several zones occur, but these occur both as (+ ■) - and (-) - mobile units. Movements of +2 cm / min were achieved with the following system:

Example composition of the solvent

electrical

Parameter

(stabilized)

15 ml. Methoxyethoxy-ethanol + 15 ml cycl. 1,2-propanediol carbonate + 13 ml Ethylene carbonate + 3 drops of nitric acid (1:30).

6 kV / mA Co ^z (chloride) 2-3 zones (+ ^{and -) Ni 2} (chloride) 2-3 zones (+ and -) Cu ² (chloride) 4-5 zones (+ and -) 6 min run time

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The medium of Example 7 also provided excellent mobility for salts of europium, lutetium, thallium and ytterbium. The location, mobility and character of the preserved zones are essential for the Material characteristic within the system under given conditions. So in the following system gave nickel and cobalt (as chlorides) 1 or 2 zones, while the mixture 3 zones accordingly the individual metal components. In addition, the zones had three colors with significantly different pink and Blue.

Example composition of the solvent electrical

Parameter (stabilized)

8 21_ml cycl. 1,2-propanediol carbonate 7.8 - 7.6 kV /

+ 9 ml methoxy-ethoxy-ethanol + 8 ml 1.25 mA

γ-butyrolactone + 3 drops of nitric acid, ^{6 min running} time (1:30).

The groups of rare earths as well as hafnium and zircon are the elements that are most difficult to dissolve. just as hafnium and zirconium form a particularly close pair, three major sub-groups are known within the rare earths. The following systems are for the transition heavy metals, Rare earth salts and zirconium / hafnium elements, e.g. those with an atomic number of 21 and above, including useful.

Example composition

of the solvent electrical

Parameter (stabilized)

15 ml cycl. 1,2-propanediol carbonate + 15 ml methoxy-ethoxy-ethanol ■ + - 13 ml ethylene carbonate + 3 drops nitric acid (1:30)

9.6 - 7.2 kV /

1.25 mA

3 min running time

In a system with cyclic! Propanediol carbonate, nitric acid, Methoxy-methoxy-ethanol and tetrahydrofurfuryl alcohol in

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In proportions similar to the above, the dye saframine moved 0 rapidly at 500 V and 100 uA, and an orange impurity remained immobile. This is an example of separating components by reaching the threshold for a Compound in the mixture.

Acidification with an inorganic acid is not essential, as the following example shows:

Example composition of the solvent electrical

Parameter - - - (stabilized)

10 12 ml 1,2-propylene glycol + 3 ml Di- 14 kV / 1.5 mA chloroacetic acid + 16 ml ethoxy-ethoxy- ■ ethanol.

Also bases such as triethanolamine or γ-picoline instead of Acid-containing media have the ability to separate metals.

The applicability of the invention to organic compounds will be through the following systems for separating sulfa drugs, Sulfamerizine, sulfaquanidine, and sulfamethazine, further illustrated.

Example composition of the solvent electrical

Characteristic '. - - - -. ^: (stabilized)

11 20 ml methoxy-ethoxy-ethanol + 12 ml 5.8-5.0 kV / l-methyl-2-pyrrolidinone + 0.8 ml di-1.25 mA chloroacetic acid. 4 min running time

The latter system, though found slow, did was to provide different zones with the dye family of rhodamine 5G, 6 G and B as well as a mixture.

The following media gave high resolutions of the above

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y? -

Dyes in 20 to 25 seconds and mobilities over 12 cm / min.

Example compositions of the solvent electrical

Parameter _s (stabilized)

24 ml cycl. 1,2 propanediol carbonate + 12 ml of ethylene diacetate + 6 ml of salicylaldehyde + 3 drops of nitric acid (1:30)

13.2 kV / 0.8 mA

24 ml cycl. 1,2-propanediol carbonate + 12 ml of ethylene diacetate + 6 ml of salicylaldehyde + 2 ml of ammonium bromide (saturated solution of methoxyethoxyethanol) + 2 ml of tributylphesphate + 4 drops of tetramethylammonium hydroxide (about 25 % in methyl alcohol)

13-12.6 kV / mA

It should be noted that urea or propylene glycol in such systems in concentrations of up to several moles do not change the conductivity, although the mobility of protein molecules does too is promoted. These substances act as thinning or suppressive agents and are in aqueous solutions for biochemical separations of substances such as proteins and enzymes are useful. The mobility of albumin in such systems can exceed that of colourants soluble in glycol, as shown by the following information on the migration from the starting point shown:

Example composition of the solvent

16 ml tris chloride (0.03 m) + 40 ml propylene glycol + 50 ml glycerine

electrical parameter (stabilized)

6 kV / 2 mA Whatman No. 1 albumin 1-1 / 4-1-1 / 2 "

soluble dye 3/4 "6 min running time

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As discussed below, the foregoing systems may be utilized in terms of speed and degree of resolution of starters, suppressive and / or stabilizing Funds are improved.

The inventive method was also carried out so that a sample was added to a bed of gel made from agar, silica and gelatin. This procedure was used to separate dyes, proteins and other types of organic compounds. The media and electrical Parameters were similar to those described in the previous examples. Separations in bulk were also made in a column carried out with powdered minerals or cellulose carriers. -

A very useful system for non-polar substances that dissolved isomers of methylnaphthalene and good dissolution for Rhodamine B and 6 G as well as food colors supplied is:

21 ml cycl. 'Propylene carbonate 9 ml methoxy-ethoxy-ethanol 12 ml tetrahydrofurfuryl alcohol 3 drops nitric acid (1:30)

The above formulation has used various types of compounds to serve various important functions Made use of. For purposes of illustration, a number of them are selected to be grouped into various ones Categories according to some of their common functions in recipes. However, these categories are not rigidly defined limits on the use of the compounds, and some fall simultaneously into several categories across their borders. So dimethyl phthalate is an example of a good one suppressive agent, although it also acts as an inert base when used as a base medium. It can go further

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insolubilizing or restricting mobility or influencing other factors in order to increase the dissolution. Water is a fairly active solvent with moderate proton donor capacity and a high dielectric constant useful. However, water is generally the main component at higher voltages in systems without external cooling less useful because of its low boiling point.

Table I Basis for the inert medium

Characteristic:

Minimum conductivity solvent, inert carrier, solution limiting

p-cymene

Mineral oil n-decanol 1-octanethiol Xylene

Inhibitors (suppressive agents)

Characteristic:

negative influence on conductivity.

Tributyl phosphate dimethyl phthalate triacetin 2-ethyl-hexyl chloride

Basis for the neutral medium

Characteristic:

low or sparse conductivity with a tendency to actively change conductivity with dilution with solvent, highly effective means of promoting solubility, coupler.

-Butyrolactone

cycl. 1,2 propanediol carbonate Propylene glycol 2-phenoxy-ethanol 2-ethyl-1,3-hexanediol Tetrahydrothiophene-1,1-dioxide methoxy-ethoxy-ethanol

Conductivity agent:

Perchloric acid dichloroacetic acid Formamide

Ammonium bromide pyridazine iodide nitric acid Mercaptoacetic acid

Basis for the active medium

Characteristic:

low conductivity with a tendency to be the conductivity of the base for the neutral medium to increase.

Strong solubilizer, medium

2 — Chloracetamide Dimethylfο rmami d N, N-dimethylacetamide 1-methyl-2-pyrrolidone

Solution

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Dimethyl sulfoxide cyclic ethylene carbonate 2,5-hexanedione

Modifying Agents

Isophorone

Nitrobenzene

Salicylaldehyde 4-hydroxy-4-methyl-2-pentanone Ethylene diacetate γ-picoline

o-dichlorobenzene

Very active media

Characteristic:

strong influence on conductivity, Proton donor solvent action and acidity-alkalinity

Diethyl ethyl phosphonate N-cyclohexyl-2-pyrrolidone Bis- (2-methoxyethy1) -ether, hexamethylene-phosphoric acid trxaemide Aminoethyl-piperazine Imino-bis-propylamine 2,2'-imino-diethanol 2-aminoethanol triethylenetetramine triethanolamine Mercaptopropionic acid mercaptoacetic acid

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A starting point for developing and choosing a solvent medium for particular chemical substances is the Determination of those media which stabilize or are compatible with the substances and which have a good to excellent result Partition coefficients in a standard chromatographic technique for the substance on which at different pH values have substrate to be used. Then the conductivity value is added for use in this procedure of the solvent as the main component set to a compatible basic system of the medium, the one in suitable Wise adjusted conductivity has, or the conductivity of the solvent can be measured so that by use of the types of agents described in Table I, a basic system for the medium is established. Mobility is normal reached at about 1.25 mA, a value which generally exceeds most threshold current values. Another Adjustment may be necessary to get the mobility or mobility of the particles of chemical substances going set or refine by adjusting the composition of the system as indicated above. For example adjustment can be made by using complexing agents, modifying agents, similar solvents, as determined by chromatographic separation, by pH adjustment and less active substrates (such as polytetrafluoroethylene).

The following list of substances can be seen in three main categories as indicated below. Other Factors to consider are a larger range of the liquid and the dielectric constant, low viscosity, Compatibility and miscibility with water and strong donor-acceptor influence or neutrality:

1. The main group has boiling points at or above 160 ^° C. and is liquid at or near room temperature. In general

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it has a good solvent effect.

2. A number of useful compounds have boiling points of less than 160 C or melting points slightly above room temperature. They are often used in smaller percentages used in modified systems. Often they can also be liquefied with a smaller amount of cosolvent or used in melting systems.

3. The remaining compounds are modifying agents, the melting points of which can be considerably higher and which are in Solution can be used with other media or in melting systems.

Based on physical characteristics, chromatographic and solvent separation experiments and the media adjusting techniques described herein can be a long series suitable compounds are developed from known commercially available materials or materials available by synthesis to those skilled in the art will. Therefore, only a brief exemplary compilation of such compounds is given below for the purpose of illustration specified:

Aldehydes, ketones, nitriles

Cinnamaldehyde hexachloroacetone

1,2-cyclohexanedione iminodiacetonitrile

4-anisaldehyde 2,4-imidoxolidinedione

p-Chlorophthetol 2-Hydroxy-acetophenone

Butyrophenone 'p-methoxyphenyl acetonitrile 4-chloro-3-hydroxybutyroni - O-methyl-anisole

^tr "il 2-methylpiperazine-N, N'-di-

carboxaldehyde

Ethyl cyanoacetate

Glutaronitrile N-morpholino-carboxaldehyde

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Nicotine Nitrile 5-Nitrosalicylaldehyde Nicotinaldehyde picolin nitrile 2-piperidone pimelonitri1 Piperonal

1,4-piperazine dicarboxyaldehyde

Pyrrole-2-carbaldehyde Pyridine-3-carbaldehyde Tetrahydroionone Veratraldehyde Alcohols, Glycols, Phenols _r Polyglycols

2-amino-2-methyl-1-propanol Cedrol

3-chloro-1,2-propanediol Cyanoethy1-sucrose Dehydro i s ophyto 1 Dehydrolinalool dichlorotriethylene glycol 2,6-dinitrothymol Furfuryl alcohol 3-hydroxycamphor hydroxyethylpiperazine

5-hydroxy-2- (hydroxymethyl) -4H-pyran-4-one

5-indanol dl-menthol 2-mercaptoethanol N-MethyIo1-2-pyrrο1i don 2,2 ', 2 "-nitrilo-triethanol 2-nitro-1-propanol

Polyethoxylated lanolin (5+) alcohols 4-pyridine-propanol 2,5-tetrahydrofuran-dimethanol

1,1, l-trichloro-2-methyl-2-propanol (and hydrates)

Thiodiethanol

Darken amides and related compounds

urea

N-ethyl-p-toluene-sulfonamide N-2-hydroxyethyl acetamide methane sulfonamide N-Athy1fοrmami d

Pieramid

Thioacetamide

2-furamide

Formamidine Acetate 4-Acetamido-butyric acid

2-acetoacetamido-4-methylthiazole

Anthranilamide

Chloral formamide

Diacetamide

N, N'-diallyl-tartaric acid diamide n-butyramide

Cinnamic acid

N-methyl-nicotinamide, N-isopropyl-salicylamide 5-hydroxy-valeramide N-methyl-propionamide

Iso-nipecotamide,

2-hydroxy-ethoxy-acetamide N-hydroxy-acetamide 3, 5-dinitrobenzamide N, N-dimethylacetoacetamide

N, N-diethyl-1-piperazine carboxamide

70 * 0847059 $

- 2 <f-

N-ethylacetamide hexamethyl-phosphoric acid

triamid
Hexamethyl phosphorous acid

triamid

N-formyl-hexamethyleneimine 2,2,2-trichloroacetamide P-nitrobenzamide sulfamide

N-FuIfonyl-stearamide Thionicotinamide pyrazinamide 2-hydroxy-ethycarbonate l-naphthalene acetamide tert-butyl carbazate lactamide

N-methyldiacetamide P-acetamidobenzaldehyde

It-ter

Butyl chloroacetate bis (2-chloroethyl) carbonate Butyl stearate 2-chloroethyl chloroacetate ethyl methyl carbamate Glycol dimercaptoacetate, hydroxyethyl acetate Iso-pentyl nitrite phenyl carbonate Methyl oleate

Polyethylene glycol (200) dibenzoate

Tetrahydrofurfuryl oleate Tributyl borate tributyl citrate

2,2,4-trimethyl-1,3-pentanediol diisobutyrate

ether

Dibenzyl ether

Dimethoxy-tetraethylene glycol Dimethy1-polysiloxane Diethoxy-ethylphthalate N-Butylphenylather Crown ethers (Crown ethers) o-Ethoxyphenol
1-ethoxy-naphthalene glycerol dimethyl ether p-hexyloxyphenol methoxyethyl ricinoleate 2-vinyl oxyethyl ether

Different connections

Lewis acids, bases and salts Tetranitromethane N-Acetyl-morpholine ■ Acriflavine

amino acids

Benzothiazole

o-aminophthalic acid hydrazide butyl sulfone

Bis (2-ethylhexyl) hydrogen phosphate

acidic dimethyl pyrophosphate guar gum

1-nitrosopiperidine indole

Pyridazine

Trimethylsulfoxonium iodide

7098847 0593

- vT-

inorganic salts, acids

Cedren

Trimethylene sulfide

Chloropicrine

Lithium stearate

inorganic salts

Calcium boron gluconate

Aminophosphoric acids

Diphenyl selenide

Bismuth ethyl campherate

Sulfur iodide

Trimethyl sulfoxonium iodide

Trichloromethyl phosphonic acid

Picric acid

Aminoethane thiolsulfonic acid

As part of the method that can be used to classify the materials as listed here these can be titrated with distilled water and their conductivities preserved. The dilution / conductivity curve obtained in this way gives the rate of change of conductivity with dilution and the point of decrease or Plateau values of conductivity as achieved in reasonable diluents, which helps improve the materials in terms of the various categories discussed here, such as being very effective solvents, effective solvents, etc. The following information illustrates the application of this technique (the resistors are given in MQ). The very low resistance plateau at the specified dilution values shows that dichloroacetic acid-r acid and mercaptoacetic acid belong to the category of very effective media. The slightly higher plateau of ethylene carbonate and 2,5-hexanedione brings this into the active media category. Such a method allows a reasonable assessment

709884/0593

ting scale for evaluating the various media be .. This technique helps. Media to a desired conductivity value by determining the resistance values to bring at different dilution values.

Sample initial resistance with resistance

stood 0.3 ml 0.05 ml water with 1 ml

addition of water

Dichloroacetic acid 100 0.042 001 Mercaptoacetic acid 1 0.050 0.004 Ethylene carbonate 0.2 0.20 0.065 ' 2,5-hexanedione 7th 2.2 0.060

Often even partial miscibility with water is sufficient, the indicate the expected area of activity or the properties. Further these examinations are countered by titration other substances than extended against water. For example, dichloroacetic acid, formamide and thiodiethylene glycol have been used. These then embody a different solvent miscibility ability and profile. Among these agents, the formamide has a very high dielectric constant and a larger one Conductivity as water, while the conductivity of thiodiethylene glycol was in the range of the water used and also reached the conductivity value of the water when a drop of water was added; i.e. after only minor Dilution with water. The changes in conductivity caused by dilution with non-aqueous substances were further characterized by adding a very small amount of secondary solvents to the water evoked changes in the plateau values were observed. This helps establish a relationship between the Influence of secondary solvents, such as the active or very active type (or the inert type for the suppressive Activity) and the conductivity profile. Such effects are secondary to the dilute means to which the

70988470593

Diluent is added, variable or characteristic. You can also use the conductivity titration curves a particular diluent of value for conductivity must be studied, which is an already ionized. system or a system with higher conductivity. E.g. the dilution of hydroxy compounds and ethers with moderate conductive aqueous ammonium nitrate solution and acetic acid. The comparison was made where both of the latter systems had equivalent conductivities. The dilution of the aforementioned aqueous conductive solutions by the compounds 1,3-butanediol, 2,2-methoxy-ethoxy-ethanol, 2-oxydiethanol. Bis (2-methoxyethyl) ether, sorbitol (40% by volume solution) and sorbitol (57% by volume aqueous solution) generally show a similar drop in conductivity over the titration range, although certain clearly defined curve shapes have been derived. So were the relative activity and the suppressive profile of the various diluents can be seen. Using this technique is the main difference easy to recognize with bis (2-methoxyethyl) ether. Differences were also found in the effects of aqueous sorbitol at different concentrations compared to non-aqueous materials to the ionized ammonium nitrate solution

which were otherwise less pronounced than on dilute acetic acid solution. The various Systems are examined in terms of how they provide the data for equilibrium characteristics, ionization and / or formation for the substances of interest and at different pH values. A large anthology or library of data can be created for these various possibilities in order to provide a scaled-down empirical basis for that To achieve conditions of system selection for use. The result of the invention is an extensive one that has already been worked out Table for basic solvent systems available from the future classification to the development of Media for use with particular species or substances

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2638393

can be done.

As by the examples above and the listing of the chemical Illustrating substances, part of the process according to the invention involves separating or making chemical substances mobile Particles or matter, conveniently on a support such as filter paper, in a medium of less Conductivity to which a high voltage is applied. The base or foundation of the medium comprises one or more compounds, e.g. inorganic or organic compounds such as glycols, ethers, esters, diones, lactones, amides, nitriles, Alcohols and water. An agent for adjusting its conductivity can be added to the medium, which is selected from the group Water, acids, bases and salts can be chosen. The voltage used in the process is in the range of about 50 up to 25,000 V / cm. At very high voltages, and especially in the case of volatile or gaseous substances, cooling may be necessary be. The preferred range is about 200 to 3000 V / cm, and in this range the process can be performed without external cooling be performed. The conductivity of the medium is preferably adjusted so that a current density in the range

2
of about 0.2 to 100 uA / cm, based on the area of the substrate, such as filter paper. The preferred area

2
is 1.4 to 54 uA / cm. For work in bulk or in substance

and with external cooling, current densities above 100 uA / cm can be used will. After its conductivity has been suitably adjusted, the transport medium has a sufficiently high voltage at a low current value (around the threshold value) exposed to the separation of the chemical substances or particles in it a speed of about 1 cm / sec to about 0.25 cm / min to induce. In the above examples, no external cooling was required under the specified conditions.

Refinement of the media compilation techniques can be lead to an improvement in the resolution in the separation of given components by EMB. E.g. * enables the medium to be selected

70388470593

5 ml of cyclic propanediol carbonate, 5 ml of propylene glycol, 2 ml of N-methylacetamide and 0.4 ml of tetrahydrofurfuryl alcohol the dissolution of Rhodamine B and 6G of Example 12 in considerably less than that required in Example 12 3.6 cm. The use of improved resolution to shorten the separation distances makes it possible to reduce the diffusion effects minimize.

It is possible to generalize the resolution according to the following To improve the procedure. A suitable solvent is found for the chemical substance to be transported. The nature the chemical substance to be transported is then analyzed in terms of its proton donor-acceptor properties. The donor-acceptor properties of a number of chemicals are listed in the literature in the catalog. See e.g. V. Gutmann, Coordination Chemistry in Non-Aqueous Solutions (1968). Then a component should be added to the medium, which is involved in a proton donor-acceptor interaction involved with the chemical substance. In many cases the proton donor-acceptor properties are chemical Substance not yet cataloged or they are complex. In such cases, the type of media components that the Improve resolving power can be determined by testing the system by simply using a very strong proton donor to one sample and then a very strong proton acceptor to another. If the strong donor has mobility (Speed of movement) of the connection increases, donors of different strengths are then tested, to determine which of them will provide the greatest improvement in agility and resolution. The procedure is analogous, when the strong proton acceptor increases the mobility of the chemical. The addition of a component that works with to relate to the chemical substance or particle to be transported through proton donor-acceptor interaction seems to facilitate the initial mobility of the chemical species. The dielectric constant of the

709884/0593

Medium is then set to a moderately high value if necessary set. It may also be necessary in terms of electrical Instability of the medium due to the addition of a compensating component, as indicated in detail above, to correct.

As a further aspect of the invention it has been found that an improved composition of the semiconducting media is there for certain chemical species also enables EMI responsiveness to develop that with the naked eye at relatively low voltages and low current values, compared with the high-voltage processes described above is. Voltages below 50 V / cm and even below 20 V / cm on conventional ones Support media were used. For example, EMB transport can be achieved with current values down to about 3 χ 10 ~ W up to 1.7 χ 10 ~ W with voltages from 2 to 4 V at 1.5 to 4.2 μΑ Achieve over several inches of Whatman # 1 filter paper. This means EMB operation at potentials of several

2
mV / cm at tenths of μνί / cm. The limit in low voltage EMB is where electrical diffusivity comes into play. It has also been found that certain agents lower the threshold current value of a chemical species (chemical substance or chemical particles). Low voltages of 0.05 to 50 V / cm applied to the medium to generate

2 supply a current density of 0.001 to 0.2 or up to 4 uA / cm can be applied.

A media system is modified to be responsive to low voltage EMI and a reduction in the threshold current value in the same way that it is modified to improve resolution. In particular starters (initiators) and mobilizing agents are added. Starters are compounds that work in the direction that they lower the threshold current value of a given chemical species, and are agile agents

70988470593

26383 ^ 3

Connections that increase mobility (transport speed) of a given chemical species. There is some overlap between the classes of compounds that are useful as starters or initiators, or as mobilizing agents, i.e., some compounds act as both Starters as well as mobilizing agents.

In general, materials that are compatible with the chemical species to be transported are at a proton donor-acceptor level act, and high dielectric constant materials useful as starters and mobilizing agents. Examples of compounds that are often useful as both initiators and mobilizing agents are N-methylacetamide and_ salicylaldehyde.

With the use of starters, currents can threshold up to

2 can be set down to 0.2-0.002 or 0.001 uA / cm, and the EMI can be set at lower but still lower currents at these currents effective speeds of movement with tensions going down at 0.05 to 10 V / cm. The tension value of 0.05 V / cm set in practice practical minimum, since stress effects of this magnitude, which are inherent in the system, occurred. All in all taking into account both the high-voltage and the low-voltage EMB, the EMB voltage range can be 0.05 to 25,000 V / cm, with the performance values coming down to

-9 -5 2

1.2 10 to 5 χ 10 W / cm.

The following examples illustrate how the media are used for EMB transport according to the criteria of semiconductivity and compatibility "with chemical species, as described in connection with high voltage EMI, with starters can be modified to lower the threshold current.

709884/0593

Example compositions of the solvent

16 7 ml propylene glycol, 3 ml diacetone alcohol (mobilizing agent, also increases that

Dissolution), 2.2 ml N-methylacetamide (high Dielectric constant, acts as a starter and agitating agent), 1.3 ml Formamide (also).

17 21 ml cycl. Propanediol carbonate, 9 ml methoxyethyl-ethanol, 12 ml tetrahydrofurfuryl alcohol, 3 drops HNO ^ diluted 1: 3 with Water.

The medium of Example 16 was used to separate rhodamine dyes and also to separate vitamin B ₁₂ and sodium riboflavin phosphate mixtures.

The medium of Example 17 was used to separate Rhodamine B and 6 G over less than 0.5 cm. The tetrahydrofurfuryl alcohol increased the mobility of the connections and thus increased the molecular differences. Without this component - the two fabrics showed almost equal movement over several centimeters.

The medium of example 17 can be changed for further examples conductivity by dropwise addition of conductivity initiators or mobilizing agents (A) to obtain the electrical values (B) and the power values (C) in the table below.

709884/05 ^ 3

middle

electrical values total EMB-Lei for the EMB-running value in (on 4xlcm filter paper)

(B) (C)

Nitric acid (1:30
in water) 4th V, 4.2 μΑ 17th Ammonium bromide (ge
saturates in glycol) 10 V, 1.1 μΑ 11 Formamide 10 V, 1.2 μΑ 12th N-methyl-acetamide 2O V, 2 μΑ AO N-Methy1-formmami d 10 V, 4.1 μΑ 40 Hexamethyl phosphorus 10 V, 3.5 UA 35

acid- tr iami_d

The EMB can be used to make biochemical substances with a high threshold current mobile, such as proteins, Polypeptides, nucleic acids, steroids, lipids, lipoproteins and fatty acids. Proteins and other biochemical compounds are subject to it slight thermal and chemical changes and are usually handled in aqueous solution, often in ice-cold, buffered electrolyte solution. Water as the main component in EMB media has the disadvantage of electrolyte solutions to form and quite evaporate. The adaptation of systems with a high water content to EMB use became special Attention also paid to the application of EMB to proteins and related substances in non-aqueous media Systems. Since the activity of biochemical compounds is tied to their structural integrity and sensitivity, was a Another aim is to put together a versatile set of media that will sustain this activity.

The general technique for formulating an aqueous EMI medium for protex transport includes degradation the conductivity of water by adding a suppressive

709884/0693

V *

- 56 -

acting agent, adjusting the dielectric constant by adding a material with a high dielectric constant, if necessary, and the addition of strong and / or mobilizing agents to encourage movement of the proteins.

As stated above, it has been found that a number of compounds the conductivity of water to varying degrees suppress and thereby alleviate the problem of high conductivity in aqueous media. These connections work too as miscible protein solvents. The results of the conductivity suppression are in the form of the following example reproduced.

Example 18 -

Numerous protein-compatible solvents were combined with water (16/9 volume ratio). Pure solvents were made used when possible as the trace impurities in commercially available materials suppress the Can affect conductivity (this is illustrated by the values given for compounds (7) and (18), which are the same substance but come from two different sources). Relative values of the suppression of the Conductivity compared to the conductivity of water were:

(1) thiodiethylene glycol 2.2

(2) 2,6-dimethylmorpholine 2,6

(3) methoxyethoxyethanol. 2.6

(4) 2-pyrrolidone 2.7

Compounds that suppress conductivity are:

(5) γ-butyrolactone 3.3

(6) Sorbitol 3.8 (70 1,3-butanediol 3.6

(8) Pronylene Glycol 3.6

(9) dimethylformamide 3.6

709884/0593

2638 9 ^ 3

A group of higher strength suppressants are:

(1O) dimethylacetamide 4.8

(11) Tetrahydrofurfuryl alcohol 4.6

(12) Butoxy-ethoxy-propanol 5.0

(13) 6-hexanolactone 5.0

(14) Oxydiethanol 5.4

(15) Diacetin 5.6

The really powerful class of water suppressants can be represented by:

(16) 2- / 2- (ethoxy-ethoxy) -ethoxy-7-ethanol 8.3

(17) 1- / 72- (2-methoxy-1-methyl-ethoxy) /

-l-methyl-ethoxy / -2-propanol 10

(18) 1,3-butylene glycol 12

The choice of an appropriate suppressant solvent should reflect the effect of the suppressant on protein migration consider. Thus, the above compounds (1O), (11) and (13) can promote protein mobility, while (3), (4), (9), (14) and (18) can be less strong in this regard.

Other solvents for biochemical compounds include alcohols such as methyl carbitol, phosphonates such as diethyl ethyl phosphonate. Lactones such as 6-hexanolactone and sugars.

The compatibility of the solvent with the substrate is another consideration. If the solvent is not selected appropriately this can attack the substrate, resulting in a changed porosity, structural breakdown or the like Effects leads. A suitable choice of the solvent in the formulations of the media enables the use of e.g. ion exchange of "thin-layer" plates and films of cellulose derivatives, such as nitrate or acetate, or of agarose,

709884/0593

-Ä «

263899

Acrylamide or silica gels impregnated with EMB media are. '

Excessive use of strong suppressants can result in a system with so high an internal resistance that there is considerable resistance heating, especially where operation at a high threshold current is indicated. So is choosing a less powerful suppressant is often satisfactory. The Tc requirements of proteins and

applied substances are often in the range of 4.6 mA / 50 cm or more on a cellulose substrate, compared to 1.2 mA /

2
50 cm or less for most other connections.

Lowering the water content of EMB media, as described above, can change the dielectric constant of the medium. This change can be made by adding components in general very high dielectric constants are compensated, whereby the high dielectric constant that is desirable for EMB is restored. Examples of suitable materials to set this dielectric constant are about hydroxyethyl formamide, N-methyl formamide, formamide, N-methylacetamide and related compounds. In general are N-alkyl and N-aryl amides can be used. Frequently these compounds tend, especially if they are just of commercial purity are, to increase the conductivity of the medium, what the suppress counteracts iven mechanism. Such a contribution to the conductivity can be used to compensate for an excessively high internal resistance, caused by a strong suppressant.

High threshold values of biochemical species or substances can be achieved in an advantageous manner through the use of starters and mobilizing agents are reduced and the mobility of the EMB increased. Many proteins were found to be for the influence of proton acceptor substances in increasing

7098 * 4/05 * 3

¥ 0

mobility particularly susceptible, but were proportionate indifferent to mobilization by proton donor molecules. Carry starter or initiator substances, even if they are present in a relatively low concentration, contribute considerably to lowering the threshold current, and if they also act as mobilizing agents, then also to increase the mobility of the species. For proteins can Initiators are used to set the thresholds of

2 2?

4.6 mA / 50 cm to 3.4 to 1.0 mA / 50 cm (20 μΑ / cm) at voltages in the range of 50 to 25,000 V / cm. Typical initiator substances, mobilizing agents, and useful ones Solvents are nitrobutanol, S-acetyl-S-chloropropyl acetate, Salicylaldehyde, N-methyl-acetamide, boric acid, phenols, guaiacol, fumaric and barbituric acid, piperazine, furfural, tributoxy- ■ aethylphosphate, β, ß, ß-trichloro-t-butyl alcohol, dimethyl-1,3-dioxolane-4-methanol, 2-ethyl-sulfonyl-ethanol, tetrahydrofurfuryl alcohol, N-substituted pyrrolidones, dimethyl sulfoxide, 2,2-oxydiethanol, cyclic ethylene carbonate, tetramethylurea, Thiodiethylene glycol, 1-ethynylcyclohexanol, tetrahydro- 3-furanol, "2, 6-dimethyl-m-dioxan-4-ol-ace did and 2,5-bis (hydroxymethyl) -tetrahydrofuran, other amides, especially N-alkyl and dialkyl and hydroxy amides, other proton acceptors and buffer systems. Substances with high dielectric constants very often act as mobilizing agents or initiators.

Electrophoresis of proteins is often done in a high pH (alkali) buffer because of the dependence on isoelectric points in electrophoresis. The EMB transport of proteins, on the other hand, can be carried out in acidic media. Buffers can be made from the list of biocompatible agents given below, or they can be of the commonly used types such as Tris, Veronal, or Sorensen. In addition, more common organic acids and bases can be used. Examples of acidic buffers used for EMB transport of proteins and related substances

701884/0593

are the following:

Tetramethyl-ammonium hydroxide / acetic acid, triethylenetetramine / 2,2-oxydiacetic acid, dimethylamine / picric acid
Diethanolamine / dichloroacetic acid
Triethanolamine / dichloroacetic acid
Piperazine / dichloroacetic acid

A compilation of means for using buffer formulations with minimal impairment of sensitive biological systems is given, as is a short list other representative means which satisfactorily must be accepted in order to be generally useful for biological work. Additional criteria for the latter Group are low melting point, a good range of liquid state, water solubility, other solubility, Solvent activity, inert behavior or functionality, etc.

Table IV

Biologically compatible buffering agents and zwitterionic buffers

Cyclohexylaminoethane sulfonic acid, N'-2-hydroxyethyl piperazine

N'-2-ethanesulfonic acid

Tris (hydroxymethyl) methylaminopropane sulfonic acid

Table V

Other representative materials suitable for use in EMB media in biological systems

Acetamidocaproic Acetanilide

L- oC -acetamido-ß-mercaptopro- allantoin

pionic acid

Acetamidophenol Alantolactone

708884/0593

i-Allyl-2,5-dimetholy ~ 3,4-methylene-dioxybenzene

η-Amy1-butyrate

Anhydromethylene citric acid

B-L arabinose

Arabitol

Arachidonic acid

Benzyl acetate

1,3-bis (hydroxymethyl) urea

Bis (2-ethylhexyl) -2-ethylhexylphosphonate

Ethoxy (10-20) glucose ethyl linoleate

Ethyl laevulinate

3-ethyl-1-hexanol 2-ethyl-2-methyl-succinimide 2-ethyl-sulfonylethanol Ethylene glycol diacetate 1-ethynyl-cyclohexanol Eugenol

Farnesol

Fructose

D-gluconic acid δ-lactone glutathione

Glycerophosphoric acid

2,6,10,15,19,23-hexamethyltetracosane

3-hydroxy-2-butanone 2-hydroxybenzyl-phosphinic acid N- (2-hydroxyethyl) palmitaraid

5-hydroxy-2-hexenecarboxylic acid lactone

2,6,10,15,19,23-hexamethyl-2,6,10,14,18,22-tetracosahexene 3-hydroxy-3,7,11-trimethyldodecanecarboxylic acid, Ichthymal
Isoascorbic acid isoeugenol inositol-hexaphosphoric acid isopropy1-myristate isovaleric acid isovaleramide kojic acid
Lactobionic acid Linoleic acid Lipoic acid Methylalacetamide Methyl nicotinate γ -methyl- oc, ß-crotonolactone N-methyl-pyrrolidinone l-methoxy-4-propenyl-benzene p-methoxy-benzaldehyde p-methoxy-benzyl alcohol myristyl alcohol 3,4- (methylenedioxy) benzaldehyde Nicotinamide ascorbate Nicotinic acid monoethanolamine

2-nitro-2-propyl-1,3-propanediol

Octane carbon (caprylic) acid oleic acid oils, natural Orotic acid

N- (pantothenyl) -ß-aminoethanethiol pantothenic acid cocoa butter

15-hydroxy-pentadecanecarboxylic acid ε-caprolactam re-ε-lactone

5-hydroxy-2 (hydroxymethyl) -4-pyrone

Choline salts

2- (chlorophenyl) -3-methyl-2,3-butanediol

709884/0593

Cineol (eucalyptol) 2-pyrrolidinone

Citric acid. . Polyethylene glycol-p-isooctyl-

cC, ß-Cyclopentamethylene tetrazole ^{phenyl ether} diethyl ethyl phosphonate
N, N-diethyl isovaleramide
N, N-diethyl-m-toluamide

Steroids, natural and derived, e.g., ex-lanolin

2,2-dimethyl-1,3-dioxolane-4-methanol

2,6-dimethyl-m-dioxan-4-ol acetate

Dimethylpolysiloxane 2,3-epoxy-2-ethylhexanamide Eicosamethylnonasiloxane Ethyl-phenyl-ether O-Ethoxybenz-ainid Propoxy (10-20) glucose pentaerythritol chloral Pentaerythritol tetraacetate 3-pentanone

3-phenoxyl-1,2-propanediol Phenoxyacetic acid

Polyethylene (20) sorbitan monooleate

oC -phenylbutyramide

2-phenyl-2-hydroxy-propionamide

2-phenyl-6-chlorophenol piperidinium salts

Poly (ethylene glycol) pnonylphenyl ether

Polyoxyethylene stearate, polyvinyl alcohol, polyvinyl pyrrolidone

N-polyoxyethylene fatty acid amides ■. ■

Salicylamide Sorbic Acid Tannins

cis-terpin hydrate tetrahydro-3-furanol

Tetrahydrofurfuryl alcohol poly-ethylene glycol

3,7,11,15-tetramethyl-2-hexadien-1-ol

2,6,10,14-tetramethylpentadecane

Tetraethylene glycol dimethyl ether

Thiamine and derivatives theophylline and salts O-thiocresol Thujic acid tiglinamide tocopherols, tocols 2,2,2-trichloroethanol Triethylene glycol

3,5,5-trimethyl-2,4-oxazolidinedione

Undecylenic Acid Veratrol

Viologens

Vital stains Valerolactone Vitamins K, A and derivatives Wetting agents

6-propylpiperonyl butyl diethylene glycol ether

3-pyridine-ethanol

709884/0693

As. Media produced above in combination with water, one or more suppressing means, one or more means for increasing the dielectric constant, and one or more initiators (and / or mobilizing agents) enable the separation of proteins within about 1 min over a distance of a few centimeters on Whatman No. 1 filter paper while doing the same separation on the same substrate up to 16 h over up to 15 cm substrate when using electrophoresis.

A number of proteins and related substances are insoluble in water. For example, some are derived or conjugated Proteins as well as some polypeptides, keratins and prolamines are insoluble in water. If this problem occurs in some Cases can be overcome by using modified aqueous media, the use of non-aqueous media offers Media additional flexibility.

An alternative to aqueous EMB systems for proteins and other biochemical compounds is the use of other solvents that give the water in the proton donor number (DZ = 18.0 for water) and in the dielectric constant (DK = 81.0 for Water) are analogous. Particularly useful are cyclic ethylene carbonate (DZ = 16.4, DK = 89.1, boiling point 245 ⁰ C) and 1,2-propanediol carbonate (DZ = 15.1, DK = 69.0, boiling point 240 C). These solvents impart superior thermal stability to the composition of the medium and allow working with larger resistance and higher voltage gradients without the need for external cooling.

Certain solvents show a more intense solvent action than water for some proteins. Keratins, which are insoluble in water, can be dissolved in other solvents such as dimethyl sulfoxide. Prolamines can be dissolved in glycols, glycol ethers and certain alcohols .

709884/0593

Solvents with moderate to strong proton acceptor properties are suitable for dissolving compounds and
can even form the basis of the medium. solvent
of these types include iodine monochloride, sulfur dioxide and hydrogen fluoride. For example, anhydrous hydrogen fluoride is a good solvent for fibrous proteins that are normally insoluble in water. Collagen substances as well
Elastins and reticulins are particularly resistant to dissolving in aqueous media, while they are soluble in non-aqueous media.

The media for the EMB transport of proteins can be different
Protein solvents that are selected for specific properties, such as glycols, amides,
Ethers, pyrrolidones, lactones, sulfoxides, phenols, alcohols and phosphonates.

Suitable aqueous systems for the transport of proteins are in accordance with the table of suppressants above using a solvent / water volume ratio from 16/9 following:

16 ml of thiodiethylene glycol
9 ml of water

2 drops of ethanolamine

(Separation of protein mixtures including
Cytochrome C and myoglobin)

electrical parameter 1.8 kV / 1.2 mA

16 ml of 6-hexanolactone
9 ml of water

3 drops of ethanolamine

(revealed protein movement and dissolution)
electrical parameter 1 kV / 1.2 mA

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16 ml dimethylacetamide 9 ml H ₂ O

4 drops of ethanolamine (separation of protein) electrical parameter 1.6 kV / 1.2 itiA

The following three examples illustrate non-aqueous media representative of those used for EMB transport of human and bovine albumin, hemoglobin, cytochrome C (an enzyme), myoglobin (muscle protein), and Pancreatin have been used. In addition, proteins have been separated from blood in experiments in which the cell fragments were left in the initial state. With these last ones In separations, phenol was a useful component of the medium.

Example 19

12 ml of cyclic ethylene carbonate 6 ml of ethoxyethoxyethanol 6 ml of thiodiethylene glycol

6 drops of tris-dichloroacetic acid buffer

Example 2O

7 ml of cyclic ethylene carbonate 7 ml of ethoxy-ethoxy-ethanol 9 ml of oxydiacetic acid 1.5 ml formamide

6 drops of tris-dichloroacetic acid buffer

Example 21

10 ml cyclic ethylene carbonate 4 ml N-methyl-pyrrolidone 3 ml furfuryl alcohol 2.5 g boric acid

708884/0593

Example 21 (continued)

4 ml of 1,3-butylene glycol

16 drops of piperazine dichloroacetic acid buffer (pH 3.7) Acridine orange (fluorescent indicator)

The following example illustrates how to separate albumins and in particular EMB media applied to globulins and electrical conditions.

Example composition of the solvent electrical

Parameters (stabilized)

22 10 ml cycl. Ethylene carbonate 5.2 kV / 2.0-

4 ml butylene glycol, 4 ml methyl 3.6 mA pyrrolidinone, 2 ml formamide (Ini- Whatman No. 1 tiator), 2.5 g boric acid, 3 ml fur (10 cm) fural (pH buffer and mobilizing agent Medium), 16 drops of piperazine dichloroacetic acid pepper, pH 3.7 (pH buffer and mobilizing agent), acridine yellow (fluorescent indicator)

The same medium was used to separate cytochrome C, hemoglobin, Myoglobin, albumin, yohimbine and atropine are used.

The use of dyes which act as tracers or tracers can in some cases be desirable in order to visually follow the separation of colorless biochemical substances, see Examples 21 and 22. However, the particular dye must not interfere with the separation process itself. Bromophenol blue has commonly been used with serum proteins, but can migrate separately from the protein in the EMB. Methylene blue is often suitable for redox-sensitive materials, and glutathione, either in oxidized or reduced form, can be used to buffer against redox reactions. Saframine-type dyes bind to proteolytic enzymes and alter their solubility properties and therefore may be useful in separating them from other materials. A few

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Milligrams of an easy-to-connect fluorescent tracer, such as acridine orange, allow visual observation of many Substances, including proteins, under UV light without changing their migration properties. Other tracers, like e.g. brighteners, fluorescent couplers, and even fluorescent antibody material, can be used to track protein transport be useful. Other tracers for biochemical and other work are natural dyes, such as the flavins and Primulin. Nile blue can be particularly useful alone or in combination with other dyes under UV and daylight. Neutral red with esculin remains sensitive when diluted about 1,000 times with daylight alone. The UV dyes are too attached to locate weak positive or negative charges in biological structures. Antibodies or others Coupling tracer substances and radioactive derivatives can also be useful, e.g. Rhodamine B isothiocyanate, fluorescein isothiocyanate, p-isothiocyanato-acridine, 4-chloromethyl-lacridine, 1-ethyl-2 - / "- 3- (1-ethylnaphthol / Ϊ, 2 ^ / - thiazolin-2-ylidin) -2-methylpropenyl / -naphth / l, ^ thiazolium bromide. Other possible tracers are phenazine methosulfate, Remazol brilliant blue R, thiazolyte blue, protoporphyrin IX, citrazinic acid, quinine, lisamine, rhedamine, clevesic acid and umbelliferone.

A further aspect of the invention is the application of the formulation technology of EMB media for the production of gaseous semiconducting media, which allows controlled conduction without the need for evacuation, very high temperatures or very high voltages. The application of the formulation techniques for liquid EMB media to the formulation of gaseous media led to the achievement of high conductivity values without the need for high potential. The purpose of building up a gaseous EMI medium is to increase the conductivity value of the gas to the value of the semiconductivity or another value suitable for the desired application.

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In industry, gases are broadly based as insulators been used. The currently practiced conduction of gases mostly aims at the high dielectric properties of gases generally. The conduit usually takes place within a jacket or other controlled environment a relative vacuum using an energy source (such as a thermoelectric filament) to control the Management instead. In such devices there is the presence of Materials with lower dielectric properties are harmful or even destructive. The conductivity of gases is currently also of importance in the field of ionization or plasma state. Attempts have been made to generate electricity by moving conductive gases relative to a magnetic field to generate J Magnetogasdynamik), but it has been found to be necessary proved to use temperatures so high that corrosion of the containers occurred. Now it has become possible to be conductive Preserve gases at or near room temperature by applying the EMB, and this makes it possible to have a practical one To supply means of generating electricity.

The formulation or composition of gaseous EMI media leads to a useful scientific technique for studying the molecular properties of substances. In addition, the invention can be used for the construction of devices for controlled conduction in gases, which are used for wireless transmission (e.g. in transformer cores without winding), for investigations of light emission, for charge transport and molecule transport in gases, electrically triggered gas diffusion and for devices with low-voltage spark spacing be used. EMI media formulation can be used to modify fuel combustion systems and the fuel itself in internal combustion engines so that the ignition distance is increased (e.g. to allow the spark plug electrodes to be moved apart to a greater distance so as to reduce reliance on explosive propagation ).

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⁴ ^ "2638S ¹ Ki

Principles similar to those applied to the production of liquid semiconducting EMI media can also be found on the production of gaseous semiconducting EMB media application. Media that contain a number of components, such as 3- or 4-component systems, are necessary to achieve a considerable To change the conductivity of the gas in order to bring it into the semiconducting area. Means, their interaction enables a proton donor acceptor interaction, increased conductivity and a higher dielectric constant or relieved, are indicated.

For example, water acts as both a donor and an acceptor molecule through hydrogen bonding in the vapor phase in interaction with proton donors, which range from strong acids to alkanols (e.g. 1,1,1,3,3,3-hexafluoropropan-2-ol), and with acceptors such as amines (including pyridines), ethers, alcohols and ketones. in the In general, solvents have proven to be particularly suitable for conduction in gases, the more conductive or active they were as EMB solvents.

Mixing materials together also promotes improved Conductivity.

For example, the introduction of a few drops of triethylenetetramine continued the resistance between the electrodes with an electrode spacing of 0.5 cm and

g drops 2.5 cm above the bottom of the tube from greater than 10 Ω in air to 10 Ω. The addition of a small iodine crystal reduced the resistance to less than 8 χ 10 Ω. Addition of formamide instead of iodine gave 1.5 10 Ω, and both together lowered the resistance to less than 5 χ 10 Ω · With some media, resistances were hundreds of Ω at low voltages (5-lOV) around room temperature and atmospheric Received pressure.

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Medium cell resistance order of addition

total amount added

Amount in the mixture

Tetramethyl urine (> 100 meg Ω) 5 parts 5 parts material

+ N-methylacetamide 1.70 π ^ Ω 1.5 parts 6.5 parts

+ I ₂ 75 KΩ 1 part 7.5 parts

+ Diethylamine 18 ΚΩ 1.5 parts 9 parts

Agents useful in formulating or composing gaseous EMI media include iodine, other halogens, Amines, volatile salts, amides, nitro derivatives including nitrosyl chloride, acid chlorides, hydrazine, oxyhalides, sulfur dioxide, Hydrogen fluoride, ammonia, or other strong proton donor or acceptor molecules, their combinations and substances that release them. According to the principles of the formulation of liquid media, as modified above, from such Offer components assembled semiconducting media a controllably conductive gaseous environment even at atmospheric pressure in air. The use of high-boiling chemicals as classified for the use of liquid EMB media requires elevated temperature to produce the gas EMI effect. Use of lower boiling solvents is therefore preferable for the production of gaseous EMB media.

Voltages of, for example, 0 / l to 30,000 V / cm can be applied to gaseous EMI media, which leads to continuous conduction (instead of an arc flashover). Modifying agents can be contained in the media to stimulate arcing in the range of 110 to 30,000 V / cm, in order to make them useful for fuel systems, in order to increase the propagation properties of the ignition sparks and / or the electrothermal evaporation before ignition -

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differentiate or expand.

Another aspect of the invention is the practical application of the EMB in a gel. The gel consistency can be between are liquid and firm. Gels are generally amenable to resistance adjustment by the addition of a small amount Amount of a conductive agent with a high dielectric constant material such as formamide or another Amide or alkyl amide derivatives and, if necessary, a coupler solvent to improve miscibility. The EMB medium can be washed into the gel, or the gel can be made with the medium. One with the use of The difficulty associated with gels as an EMB medium is that by-products still remain from the gel production process must be removed if they affect the setting of the EMB conductivity and the transport.

Agar gel, polyvinyl alcohol, silica gel, starch gel, carboxypolymethylene (Carbopol) and "Crash Safe Aviation Jet Fuel" (with additives modified kerosene) are examples of gels that act as EMB media if they are used according to the principles described above with the appropriate conductivity-modifying Funds are endowed. Acrylamides could also be used. An example of one made with an EMB solvent Gels are polyvinyl alcohol made into a gel with tetrahydrofurfuryl alcohol. Tetraethyl orthosilicate, which gives a clear, glassy gel with numerous organic gels, enables compatibility with numerous organic gels EMB media. Gelatine also provides a clear gel base. Examples of chemical substances transported in such gels are about dye molecules, and even particulate Substances can be moved at high speeds in a "fluid" gel, such as the "crash safe aviation fuel" will.

As with EMB, voltage and current values are recorded on cellulose

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carriers set. A slightly higher current than 1.2 mA /

2
50 cm can also be used for thin gel plates (up to 3.175 mm (1/8 ")). Otherwise, gels of 9.53 to 12.7 mm (3/8 to 1/2") or more require careful choice of current to avoid excessive heat build-up.

In EMB separation processes, gels are capable of increased dissolution or separation due to their fine pore structure. EMI-induced movement of dye molecules in a gel can be used as an analytical technique to study the structure and properties of the gel itself.

The device used for GeI-EMB was different from that used for liquid EMB by having the filter paper substrate was replaced by the gel.

EMB in a gel is illustrated by the following examples.

Example composition of the solvent

23 glycol, ammonium bromide and formamide

24 5 w / v -% ceresin or micro wax

in 20 % or 30 % xylene plus EMB medium components suitable for use with xylene.

The components of the EMB medium mentioned in Example 24 you can use the 4-component system described on page 12 or ammonium bromide in methoxy-ethoxy-ethanol, 2 (2-ethoxy-ethoxy) -ethanol, Dimethylformamide, dimethylacetamide, dimethyl sulfoxide, be n-butanol or N-methyl-pyrrolidinone.

Electromagnetic fields can also be used for various purposes can be applied to the driving voltage in connection with the EMB. A second set of electrodes at an angle to that

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Set, which applies the driving voltage, can be used to remove the transported chemical particles from a straight direction of movement to throw. Similarly, one or more electrodes can be angled to the set that is driving Applied voltage, used to detect any slight lateral deviation or spread of a migrating on a substrate Compensate species and counteract the diffusion effect. In addition, a balanced pair of electrodes can be placed perpendicular to the path of the chemical species being transported and for detecting the passage of different zones of chemical species based on the change in electrical ones Forces can be used between the second set of electrodes.

Pulsating direct current fields can be used instead of a constant direct current driving force to reduce the heating of the medium can be used. As an additional modification, an alternating current field can be superimposed on the direct current driving force, in order to average the dielectric and semiconducting properties of the medium as well as in order to obtain from the Debye-Falkenhagen (solvent) -Effect to take advantage.

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Claims (4)

PATENT CLAIMS Method for moving chemical particles, characterized in that a semiconducting transport medium is connected to a to generate a current density in the range from about 0.001 to 2
400 µA / cm, equal to or above the threshold current value for the particles in the medium below which the particles remain essentially stationary, sufficiently high voltage of about 0.05 to 25,000 V / cm is applied to induce a high speed of movement of the particles . 2. The method according to claim 1, characterized in that a working at high voltage and low current density permitting semiconducting transport medium one for generating a current density in the range of about 0.2 to 2
400 µA / cm, equal to or above the threshold current value for the particles in the medium below which the particles remain essentially stationary, sufficient voltage of about 50 to 25,000 V / cm is applied to induce high speed of movement of the particles. 3. The method according to claim 1, characterized in that a semiconductor fluid transport medium which is a component of the group of mobilizing agents and initiators, one for generating a current density in the area from about 0.001 to 0.2 μΑ / cm, equal to or above the threshold current value sufficiently high voltage for the particles in the medium under which the particles remain practically stationary from about 0.05 to 50 V / cm for inducing a high speed of movement the particle is applied. 4. The method according to any one of claims 1 to 3, characterized in that the semiconducting transport medium is a Conducting at an applied voltage of about 0.05 to about 30,000 V / cm is obtained. 709 88 47 0 5 93