WO1994016319A1 - Analysis method - Google Patents
Analysis method Download PDFInfo
- Publication number
- WO1994016319A1 WO1994016319A1 PCT/SE1994/000025 SE9400025W WO9416319A1 WO 1994016319 A1 WO1994016319 A1 WO 1994016319A1 SE 9400025 W SE9400025 W SE 9400025W WO 9416319 A1 WO9416319 A1 WO 9416319A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- template
- capillary
- polymer
- product
- analysis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44747—Composition of gel or of carrier mixture
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2600/00—Assays involving molecular imprinted polymers/polymers created around a molecular template
Definitions
- the present invention describes a new product and its use in the analysis of components in samples.
- the product is characterized by that it contains at least one capillary
- electrophoresis system and one template- or a molecular imprinting material/polymer.
- Capillary electrophoresis (even called CE below) (Reference 1 : Wallingford and Ewing, 1989, Capillary Electrophoresis, in Advances in Chromqatography, 29, 1-76, Marcel Dekker, New York) is a relatively new technique which application area is rapidly increasing.
- the method is a miniaturization of the conventional gel electrophoresis method and includes the use of a thin fused silica capillary (10-100 urn in diameter) in which the electrolyte and the analyte are distributed between anode and cathode in a field of high voltage strength.
- the present invention describes a product and a method to minimize these problems.
- the present invention describes a product which contains at least one template polymer and a capillary electrophoresis system (See reference 1 for a general description of earlier CE-systems) to analyse an analyte in a sample and also a method for analysis in which the product according to the invention is used for the analysis.
- a capillary electrophoresis system See reference 1 for a general description of earlier CE-systems
- the template molecule (is often the same as the target molecule for the separation - the analyte) is mixed with functional monomeres (which can make ionic and or hydrogen bonds with the template molecule or organize itself with the template molecule in some other way) in a solution.
- the solution with the template- functional monomers is polymerized ( in the capillary, or outside the capillary that is used for CE) with or without cross-linker and after the polymerisation the template molecule is washed away from the resulting polymer.
- the resulting polymer can then be used in the product and in the method according to the invention.
- template polymer is polymers made from acrylate monomers with or without cross-linkers, where the template molecule (the analyte) is a chiral substance, an amino acid, a peptide, a
- the template polymer can be a polymer which have been made in a template polymerization with for example a carbohydrate or a nucleotide or a nucleotide sequence as template molecule and with monomeres containing boronate groups or containing nucleotide groups.
- the resulting template polymers can be used in a product according to the invention and be selective towards for example carbohydrates, oligonucieotide sequences or for example other analogous substances.
- the template polymer and the product, according to the invention can also be used for separation and analysis of other substances than the analyte.
- template polymers which have been prepared with a complexing agent as template molecule, for example cyclodext lines and other molecules which can be used as complexing agents in CE.
- the resulting template polymer will then be used in the product, according to the invention, for example showing selectivity towards the complexing agent which will be retarded and therefore the migration of the analyte in the system will also be influenced.
- the template polymers can be prepared with all, now and in future, available methods for preparation of this type of polymers (see for example ref. 1-5) and can be used in for instance the present and future CE-systems (see for example reference 1 above and the references 8-10 below) and this does not limit the scope of the invention.
- the polymers can be produced in non-water based systems or in systems containing more or less water. Below some examples of how the invention can be used are, but which in no way are meant to limit the scope of the invention.
- EDMA ethyleneglycol dimethacrylate
- MAA methacrylic acid
- electrophoresis electrolyte consisting of 7/3 (V/V) of acetonitrile/potassium
- Template polymer and capillary which have been made according to some of the standard principles mentioned in example 1 or 2 , but in which procedure another amino acid- or peptide derivative, a therapeutic, a monosaccharide, an oligosaccharide, a nucleoside, a nucleotide, an oligonucieotide or a derivative of one of these, or any in the description mentioned substances, have been used instead of PAM, BAM and LFA in the polymerisation.
- a template polymer can either be made in situ in the capillary or can be packed in the form of particles in the capillary.
- the capillary is thereafter used in the product, according to the invention, for separation.
- the initiation of the polymerisation can be made either with thermic or photolytic initiation.
- template molecules are amino acids, peptides, nucleotide s,
- oligonucleotides purine or pyrimidine derivative, basic compounds, amidine or guanidine containing compounds, amino alcohols, carbohydrates,
- the paniculate material according to example 5 can be any of the earlier
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention describes a new product and its use in the analysis of components in samples. The product is characterised in that it contains at least one capillary electrophoresis system and one template- or imprinted material/polymer.
Description
ANALYSIS METHOD
The present invention describes a new product and its use in the analysis of components in samples. The product is characterized by that it contains at least one capillary
electrophoresis system and one template- or a molecular imprinting material/polymer.
Capillary electrophoresis (even called CE below) (Reference 1 : Wallingford and Ewing, 1989, Capillary Electrophoresis, in Advances in Chromqatography, 29, 1-76, Marcel Dekker, New York) is a relatively new technique which application area is rapidly increasing. The method is a miniaturization of the conventional gel electrophoresis method and includes the use of a thin fused silica capillary (10-100 urn in diameter) in which the electrolyte and the analyte are distributed between anode and cathode in a field of high voltage strength.
Different types of detectors can be used, most common is UV/VIS detector but even mass spectrophotometers are used and this does not limit the scope of the invention. Compared with conventional fluid chromatography the method has many advantages:
- Smaller samples (picograms or lower) can be analysed and in small volumes of electrolyte (1-10 nl).
- Many band broading mechanisms which are present in LC are avoided and one gets extremely high resolution and efficiency.
- The procedure is suitable for automatization and
- The technique is suitable for analysis of more or less pure samples, "crude samples".
The separation is based on size and charge, but by using complexing agents, for example cyclodextrins or boronates, one can separate neutral compounds. The high plate number together with the small test volumes and the possibility to analyse crude samples make the method very attractive for analysis of complex molecules and for process control. Specific analysis of different substanses is however still a problem and the present invention describes a product and a method of analysis to address this problem.
Within high pressure chromatography, materials based on polymers prepared with the help of molecular imprinting (MA)1 -5, also called template-polymers or imprinted polymers, has
shown to have a high selectivity and affinity towards its substrate (the print molecule, also called the template molecule) in the separation of samples containing the template molecule. Packed in a chromatography column, template polymeres have been used for separation of the template molecule from molecules of similar structure (enantiomers, diastereoisomers, positional and structural isomers). The method has however
disadvantages such as the need for a larger amount of the print molecule, water soluble substances have resulted in poor imprinting effects and furthermore a relatively high plate number with broad bands and bad resolution have often been obtained. The present invention describes a product and a method to minimize these problems.
The present invention describes a product which contains at least one template polymer and a capillary electrophoresis system (See reference 1 for a general description of earlier CE-systems) to analyse an analyte in a sample and also a method for analysis in which the product according to the invention is used for the analysis. In a typical non-limiting
procedure for preparation of the template polymer, the template molecule (is often the same as the target molecule for the separation - the analyte) is mixed with functional monomeres (which can make ionic and or hydrogen bonds with the template molecule or organize itself with the template molecule in some other way) in a solution. The solution with the template- functional monomers is polymerized ( in the capillary, or outside the capillary that is used for CE) with or without cross-linker and after the polymerisation the template molecule is washed away from the resulting polymer. The resulting polymer can then be used in the product and in the method according to the invention.
Both cross-linked template polymer particles, solid polymer as well as soluble template polymers can be used according to the invention. A non-limiting example of a template polymer is polymers made from acrylate monomers with or without cross-linkers, where the template molecule (the analyte) is a chiral substance, an amino acid, a peptide, a
carbohydrate, an oligonucieotide, or a derivative of some of these (derivative is for example a chemically or enzymatically modified substance) a medicament, a β-blocker, etc.
As another non-limiting example one can mention that the template polymer, according to the invention, can be a polymer which have been made in a template polymerization with for example a carbohydrate or a nucleotide or a nucleotide sequence as template molecule and with monomeres containing boronate groups or containing nucleotide groups. The resulting template polymers can be used in a product according to the invention and be selective towards for example carbohydrates, oligonucieotide sequences or for example other analogous substances.
The interaction between the analyte and the template polymer in the product, according to the invention, in the CE-system influences the migration of the analyte in the capillary and the product and the method according to the invention can be used for analysis of a chiral substance, an amino acid, a peptide, a carbohydrate, an oligonucieotide, or a derivative of some of these, a medical substance, a β-blocker, etc. The template polymer and the product, according to the invention, can also be used for separation and analysis of other substances than the analyte.
Other non-limiting examples are template polymers which have been prepared with a complexing agent as template molecule, for example cyclodext lines and other molecules which can be used as complexing agents in CE. The resulting template polymer will then be used in the product, according to the invention, for example showing selectivity towards the complexing agent which will be retarded and therefore the migration of the analyte in the system will also be influenced.
The template polymers can be prepared with all, now and in future, available methods for preparation of this type of polymers (see for example ref. 1-5) and can be used in for instance the present and future CE-systems (see for example reference 1 above and the references 8-10 below) and this does not limit the scope of the invention. The polymers can be produced in non-water based systems or in systems containing more or less water.
Below some examples of how the invention can be used are, but which in no way are meant to limit the scope of the invention.
EXAMPLE 1.
To a mixture of the anti-bacterial therapeutic Pentamidine (PAM) (0.125 mmol)
in ethyleneglycol dimethacrylate (EDMA) (12mmol) methacrylic acid (MAA)
(0.5 mmol), i-propanole (2.8 ml) and water (1.3 ml) the initiator azobisiso- butyronitrile (AIBN; 12 mg) was added. Nitrogene was bubbled through and
the mixture was sucked into a fused silica capillary (ca 40 cm long). The
capillary ends were placed in the monomer solution and the polymerisation
was carried out thermically with the capillary left at 60 degrees Ceicius over
night. Thereafter the capillary was connected to a HPLC-pump with a tight
capillary ferrule and acetonitrile/water 7/3 (V/V) was passed at a pressure of
ca 1500 PSI during ca 30 minutes. Thereafter the electrophoresis electrolyte
was pumped through the capillary in the same way as mentioned above.
Using a standard equipment Beckman GoldR CE-equipment and with a
electrophoresis electrolyte consisting of 7/3 (V/V) of acetonitrile/potassium
phosphate buffer (0.05 M, pH 2), a current of ca 4.8 microampere (separation
voltage ca 5 kV) and an injected concentration of 1 mM (6 kV - 1 s) of PAM and
benzamidine (BAM) gave separation of PAM and BAM (retention times in this
example were 5.6 and 5.2 minutes). With a capillary made in the same way
but with BAM as print molecule, the retention times 5.5 resp. 6.1 for BAM
respective PAM were obtained. At pH 3 with the PAM-capiliary the retention
times 6.5 and 25 minutes for BAM respective PAM were obtained whereas the
BAM-capillary gave the retention times 6.0 respective 6.8 minutes, that is a
clear affinity for PAM was obtained with the PAM-capillary which can be
controlled with for example pH.
EXAMPLE 2.
To a homogenous mixture of the chiral imprinting molecule L-phenylalanine
anilide (LFA; 0.025 mmol) in EDMA (2 mmol) MMA (0.4 mmol) and cyclohexanol/dodecanol (4:1 ; V/V; 3 ml) AIBN was added, nitrogene was bubbled through and the mixture was sucked into a fused silica capillary (ca 40 cm long) intended for capillary electrophoresis and polymerisation was carried out according to Example 1. Thereafter the capillary was connected to an HPLC-pump with a tight capillary ferrule and ethanol was passed through at a pressure of ca 1500 PSI during ca 30 minutes. After this, electrophoresis electrolyte was pumped through the capillary and D- and LFA were injected into a Beckman CE-system.
EXAMPLE 3.
Template polymer and capillary which have been made according to some of the standard principles mentioned in example 1 or 2 , but in which procedure another amino acid- or peptide derivative, a therapeutic, a monosaccharide, an oligosaccharide, a nucleoside, a nucleotide, an oligonucieotide or a derivative of one of these, or any in the description mentioned substances, have been used instead of PAM, BAM and LFA in the polymerisation.
EXAMPLE 4.
A template polymer in which a chiral compound or a diastereoisomer or one of the substances described above or mentioned in the examples above have been used as print molecule and in which the template polymer was prepared in the capillary or packed in the capillary followed by electrophoretic
separation of the print molecule or of other compounds.
EXAMPLE 5.
A template polymer can either be made in situ in the capillary or can be
packed in the form of particles in the capillary. The capillary is thereafter used in the product, according to the invention, for separation. The initiation of the polymerisation can be made either with thermic or photolytic initiation. As
monomer one can preferably use one or some of styrenβ methacrylate,
acrylate, acryiamide, boronate or a nucleotide derivative of any of these.
Examples of template molecules are amino acids, peptides, nucleotide s,
oligonucleotides, purine or pyrimidine derivative, basic compounds, amidine or guanidine containing compounds, amino alcohols, carbohydrates,
derivatives of any of the complex forming substances that are or have been used in CE, for example boronates or cyclodextrins, derivatives of any of
these, chiral compounds, proteins.
EXAMPLE 6.
The paniculate material according to example 5 can be any of the earlier
mentioned template polymers or of the same type as these.
1 (a) Wulff. G. In Polymeric Reagents and Catalysts: Ford. W.T.. Ed.: ACS Symposium Series 308: American Chemical Society: Washington. DC. 1986. (b) Wulff. G.: Vesper. W.: Grobe-Einsler. R.: Sarhan. A. Makromol. Chem. 1977. 178. 2799. (c) Shea. KJ.: Dougherty. T. K. J. Am. Chan. Soc. 1986. 108. 1091. (d) Wulff. G.: Heide. B.: Helfmeier. G. J. Am. Chan. Soc. 1986. 108. 1089. (e) Andeisson. L.: Seliergren. B.: Mosbach. K. Tetrahedron Lett. 1984.25. 5211.
2 (a) Seliergren. B.: Lepisto. M.: Mosbach. K. J. Am. Chem. Soc. 1988. 110. 5853. (b) Lepisto. M.: Sellergren. B. J. Org. Chem. 1989. 54. 6010. (c). Sellergren. B. Chiraliry. 1989. 1. 63. (d) Andersson. L.L: Mosbach. K. J. Chromatogr. 1990. 516. 313. (d) Fischer. L.: Müller. R.: Ekberg. B.: Mosbach. K. J. Am. Chum.Soc. 1991. 113. 9358- 9360.
3 Shea. K. J.: Spivak. D. A.: Sellergren. B. J. Am. Chem. Soc. 1993. 115. 3368.
4 Vlatakis. G.: Andersson. L. I.: Mü ller. R.: Mosbach. K. Nature 1993. 361. 645.
5 Immunoassays for Trace Chemical Analysis: Monitoring Toxic Chemicals in Humans. Food. and the Environment Vanderlaan. M.: Stanker. L. I I.: Walkins. B. E.: Roberts. D. W. Eds.. American Chemical Society. Washington D.C. 1991.
6 Sellergren. B.: Shea. K. J. J. Chromatogr. 1993. 635. 31-49.
7 Wallingford. R.A.: Ewing. A.G. Adv. Chromatogr.. 1989. 29. 1.
8 Baba. Y.: Tsuhako. M. Trends in Analytical Chemistry. 1992. 11. 280.
9 (a) Chu. Y-H: Wh itesides. G. M. J. Org. Chem. 1992. 57. 3524. (b) Jelinek. I.: Snopek. J.: Smolkova- Keulemansova. E. J. Chromatogr. 1988. 439. 386.
Claims
1. Product for analysis containing at least one template polymer and one capillary electrophoresis system.
2. Method for analysis in which the product according to claim 1 is used.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE9300132A SE9300132D0 (en) | 1993-01-14 | 1993-01-14 | POLYMER FOR CAPILLARY ELECTROPHORESIS |
| SE9300132-9 | 1993-01-14 | ||
| SE9303173-0 | 1993-09-28 | ||
| SE9303173A SE9303173D0 (en) | 1993-01-14 | 1993-09-28 | Analytical method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO1994016319A1 true WO1994016319A1 (en) | 1994-07-21 |
Family
ID=26661627
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/SE1994/000025 Ceased WO1994016319A1 (en) | 1993-01-14 | 1994-01-13 | Analysis method |
Country Status (2)
| Country | Link |
|---|---|
| SE (1) | SE9303173D0 (en) |
| WO (1) | WO1994016319A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000007702A3 (en) * | 1998-08-03 | 2000-04-20 | Poly An Gmbh | Template-textured materials, methods for the production and use thereof |
| WO2001039881A1 (en) * | 1999-12-03 | 2001-06-07 | Elipsa Gmbh | Method for producing template-textured materials with high binding specificity and selectivity and utilization of said materials |
| US7397553B1 (en) | 2005-10-24 | 2008-07-08 | Kla-Tencor Technologies Corporation | Surface scanning |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0324539A2 (en) * | 1988-01-12 | 1989-07-19 | Northeastern University | Improved capillary gel electrophoresis columns |
| US5047438A (en) * | 1988-09-26 | 1991-09-10 | Supelco, Inc. | Porous rigid resins and process of preparation |
-
1993
- 1993-09-28 SE SE9303173A patent/SE9303173D0/en unknown
-
1994
- 1994-01-13 WO PCT/SE1994/000025 patent/WO1994016319A1/en not_active Ceased
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0324539A2 (en) * | 1988-01-12 | 1989-07-19 | Northeastern University | Improved capillary gel electrophoresis columns |
| US5047438A (en) * | 1988-09-26 | 1991-09-10 | Supelco, Inc. | Porous rigid resins and process of preparation |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000007702A3 (en) * | 1998-08-03 | 2000-04-20 | Poly An Gmbh | Template-textured materials, methods for the production and use thereof |
| US6670427B1 (en) | 1998-08-03 | 2003-12-30 | Poly-Am Gmbh | Template-textured materials, methods for the production and use thereof |
| WO2001039881A1 (en) * | 1999-12-03 | 2001-06-07 | Elipsa Gmbh | Method for producing template-textured materials with high binding specificity and selectivity and utilization of said materials |
| US6753396B2 (en) * | 1999-12-03 | 2004-06-22 | Elipsa Gmbh | Method for producing template-textured materials with high binding specificity and selectivity and utilization of said materials |
| US7397553B1 (en) | 2005-10-24 | 2008-07-08 | Kla-Tencor Technologies Corporation | Surface scanning |
Also Published As
| Publication number | Publication date |
|---|---|
| SE9303173D0 (en) | 1993-09-28 |
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