US3912655A - Reagent impregnated paper strips for use in preparing TLC spray reagents - Google Patents
Reagent impregnated paper strips for use in preparing TLC spray reagents Download PDFInfo
- Publication number
- US3912655A US3912655A US428565A US42856573A US3912655A US 3912655 A US3912655 A US 3912655A US 428565 A US428565 A US 428565A US 42856573 A US42856573 A US 42856573A US 3912655 A US3912655 A US 3912655A
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- US
- United States
- Prior art keywords
- reagent
- visualization
- spray
- drug
- impregnated
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- 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
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/282—Porous sorbents
- B01J20/285—Porous sorbents based on polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2220/00—Aspects relating to sorbent materials
- B01J2220/40—Aspects relating to the composition of sorbent or filter aid materials
- B01J2220/49—Materials comprising an indicator, e.g. colour indicator, pH-indicator
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2220/00—Aspects relating to sorbent materials
- B01J2220/50—Aspects relating to the use of sorbent or filter aid materials
- B01J2220/54—Sorbents specially adapted for analytical or investigative chromatography
Definitions
- Thin-layer chromatography has gained wide acceptance in drug screening procedures, since the analysis can be performed rapidly, meets sensitivity and specificity requirements, and can be used to encompass a wide spectrum of drugs and their metabolites differing both in chemical structure and pharmacological action.
- a good summary of thin-layer chromatography can be found in U.S. Pat. No. 3,590,006.
- Thin-layer chromatography screening procedures for detecting drugs in urine may be found in, e.g., Davidow, B. et al., American Journal of Clinical Pathology, Vol. 50, No. 6 pp. 714-719 (1968); and Weissman, N. et al., Clinical Chemistry, Vol. 17, No. 9, pp. 875-881 (1971).
- TLC drug screening procedures if the components separated on the TLC plate then viewing it under ultraviolet light.
- Other known developing procedures may also be used such as heat or developing agents, e.g., iodine vapor, acids, alkalis, dyes, etc., the purpose being to render the separated components on the TLC plate distinctly visible.
- TLC chromatographic visualization reagents useful for preparing spray reagents are premeasured, absorbed and dried on suitable paper strips ready for instant use. Essentially all that need be done to prepare the spray reagent is to dip the dried paper strip in the appropriate solvent. In such a manner, under proper storage in foil or containers, the shelf life of spray reagents can be increased indefinitely. Fresh reagents can be prepared in a matter of minutes, e.g., 2 to 5 minutes. The weighing of small amounts of chemical reagents by the individual carrying out of the test is avoided.
- reagent impregnated strips of the present invention specific reagents and amounts are provided in a stable form on paper strips which can be easily eluted and used for chromatographic development in the determination of drug screening by thin-layer chromatography.
- the impregnated paper strips deliver premeasured reagents in dry condition for better stability. They reduce shipping weight and cost of the reagents to the purchaser thereof.
- the reagent impregnated strips of this invention are not indicator strips such as the conventional pI-I indicator and diabetes test strips. Instead, they dispense small amounts of preweighed visualization reagents in stable form. The reagent is evenly spread over theentire area of the strip on both sides and thus the reagent dissolves or is eluted in the appropriate solvent with ease.
- diagnostic paper test strips containing various reagents which change color on exposure to blood, urine, serum, etc., pH indicator strips and diabetes test strips are, of course, well known.
- U.S. Pat. No. 3,598,533 discloses a diagnostic paper strip applicable to whole blood samples of humans in testing for various disease states and is prepared by dipping a diagnostic paper strip into an organic solvent solution of cholesterol, resin and their esters and drying the strip. In use, the treated strip is dipped directly into blood and kept for a period of time long enough to permit color development on the strip. In contrast, the present invention is not concerned with color development on the strip.
- 3,418,079 discloses a diagnostic paper test strip for the determination of protein in biological fluids comprising an absorbent carrier impregnated with a dyestuff indicator, an acidic buffer and an inorganic sulfate.
- the principal involved is merely one of color change or development on a strip.
- U.S. Pat. No. 3,016,292 is concerned with a color test indicator for detecting glucose with comprises a closed purpose is for the measurement of a specific area of the paper through elution and colorimetric determination of the dye contained therein.
- the paper strips of the present invention contain a specific amount of chemical reagent capable, upon elution, of reacting with a drug (e.g., amphetamine) to produce a colored spot on a paper chromatogram containing the drug and thus providing qualitative evidence of the presence of the drug in the uring being analyzed.
- a drug e.g., amphetamine
- U.S. Pat. No. 2,992,971 biological stains are used to prepare microscope slides of cells but such does not perform a reaction with another compound (e.g., narcotic).
- the medium used to stabilize the stains is one of the celluloses and a solubilizing compound such as urea must also be present.
- U.S. Pat. No. 3,] 38,544 discloses antibiotics contained in compressed discs composed ofa mixture of cellulose materials. The discs are then placed on an inoculated agar plate to determine zones of inhibition.
- the reagents of the present invention are eluted from their carriers prior to being applied to the TLCplate or strip as a spray reagent to produce a chemical reaction resulting in color formation on the plate or strip which qualitatively identifies a particular drug.
- U.S. Pat. No. 3,666,421 discloses a single test slide or card which contains reagents for performing immunochemical reactions such as blood group typing, pregnancy tests, etc. In particular, the reagents are adsorbed antigens or antibodies.
- Pat. No. 3,678,151 discloses a method for staining a biological specimen on a slide for microscopic examination.
- a reagent strip called Gugol Blue Stain is marketed by Wampole Laboratories, Stamford, Connecticut. This strip is used for staining a blood smear slide for differential diagnosis of white blood cells. However, instead of soaking slides in a stain solution, a strip containing staining material is placed over a slide.
- U.S. Pat. No. 3,625,652 discloses an apparatus and TLC method for the detection of narcotic analgesic and amphetamines using Amberlite XAD-2' resin to adsorb the narcotic analgesics and amphetamines from human urine.
- the U.S. Pat. No. 3,625,652 discloses collecting the urine fractions adsorbed on Amberlite XAD-2 resin and analyzing the fractions by thin-layer chromatography. As disclosed in the U.S. Pat. No. 3,625,652, the thin-layer chromatography plates are sprayed with a reagent specific for visualizing the drugs, i.e., narcotics and amphetamines.
- the U.S. Pat. No. 3,625,652 notes that the iodoplaninate reagent used was prepared according to the directions of Cochin and Daly, Vol. 18 Experientia, p. 294 (1962), by adding 250 ml. of a 4% solution of potassium iodide to ml. of a 10% solution of platinum chloride and diluting with 500 ml. of water.
- the present invention one can use the already prepared dry potassium iodide and chloroplatinic reagent impregnated strips to prepare the iodoplatinate spray reagent used in the TLC procedure disclosed in the U.S. Pat. No. 3,625,652 by merely placing the strips in the appropriate solvent.
- the invention here resides in the concept of a dry chemical visualization reagent impregnated bibulous material, e.g., paper strip, ready for instant use in preparing a spray reagent solution for drug detection by thin-layer chromatography comprising a paper strip containing impregnated therein the dried residue of a liquid solvent solution containing a visualization reagent.
- a dry chemical visualization reagent impregnated bibulous material e.g., paper strip
- the invention also resides in a method of preparing a dry chemical visualization reagent impregnated paper strip for use in preparing a spray reagent solution for drug detection by thin-layer chromatography which comprises dipping a paper strip into a liquid solvent solution of a visualization reagent.
- the invention resides further in a method of preparing a chemical visualization spray reagent solution for drug detection by thin-layer chromatography which comprises dipping an impregnatedpaper strip into the appropriate spray reagent solvent solution.
- chromatography detection agents or visualization reagents which can be premeasured, absorbed and dried on paper strips as set forth herein.
- paper strips have been impregnated with the spray reagents ninhydrin; diphenylcarbazone, chloroplatinic acid and potassium iodide (the lattertwo used together to make the spray reagent, iodoplatinate).
- Ninhydrin is and has been used for the detection of free amino and carboxy groups (e.g., amphetamine and other primaryamine compounds), diphenylcarbazone in the identification of barbiturates, and chloroplatinic acid and potassium iodide (as iodoplatinate) in the identification of alkaloids.
- the liquid solvent solution in which the visualization reagent is placed and the unimpregnated strip is dipped can be water, methanol or other suitable solvents which do not adversely affect the visualization agent and those in which it is soluble.
- Suitable spray reagent solvent solutions in which the impregnated strip is placed for elution are acetone, butanol, water, methanol as set forth herein and as known in the art, e.g., Bastos, et al., Clinical Chemistry, Vol. 16, No. 11, pp. 931-940 (1970).
- the impregnated paper strips of the present invention contain a specific amount of chemical visualization reagent capable, when eluted in the appropriate solvent, and sprayed on the chromatographic sheet, of reacting with the adsorbed drugs or their metabolite to produce a characteristic color spot for that particular drug or metabolite and thus provide clear'visual evidence of the presence of the drug in the donors urine being analyzed.
- bibulous material e.g., paper
- suitable weight and absorptivity is
- the strips are then dipped in the desired concentration of visualization remade as described herein, is removed from its package, foil or container, and placed in the desired amount of appropriate spray reagent solvent.
- the impregnated reagent is eluted from the strip into the solvent to give a spray reagent solvent containing the amount of visualization reagent on the strip.
- the spray reagent solvent is ready to be sprayed on the chromatographic plate for visualization purposes in manners known in the art, e.g., Davidow, B. et al., Am. J. Clin. Pathol. 38: 714 (1968).
- Suitable spray equipment for maintaining and dispensing the spray reagent to the TLC plate such as chromatographic spray bottles, etc., are known and readily obtainable.
- chromatographic spray bottles, etc. Upon spraying the reagent on the TLC plate, the particular characteristic color for that reagent and its corresponding drug, drugs or metabolites will appear on the plate, if present.
- paper has been impregnated with visualization reagent using paper weighing 185 g./mm with medium flow rate, smooth surface and with a thickness of 0.33 mm.
- Whatman chromatography paper 3MM meets the above criteria and it is this paper which has been used.
- the initial size of the Whatman paper was 46 57 cm. Strips were cut 5 cm. X 57 cm. prior to impregnation with reagents.
- the visualization reagent to be impregnated was placed in the appropriate liquid solvent containing the desired concentration of reagent and the reagent solution kept in a shallow container, e.g., watch glass or petri dish. The 5 cm. X 57 cm.
- EXAMPLE 3 Use of Spray Reagent Solution Prepared from impregnated Paper Reagent Strips in TLC for Drug Detection A. Ninhydrin Spray Reagent Solution This example illustrates the use of a strip impregnated with ninhydrin for the detection of amphetamine in thinlayer chromatography.
- Urine Toxicology Control Drugs 1 (Analytical)? 7.
- Mercuric Sulfate Reagent (0.25% in 10% v/v One vial pooled human urine, freeze dried. containing 0.125 mg. of amobarbital, 0.125 mg. secobarbial and 0.125 mg. of phenobarbital, reconstituted with 25 ml. distilled water. Available commercially from Lederle Diagnostics, American Cyanamid Company, Pearl River, New York.
- Urine Toxicology Control Drugs l (Analytical) One vial. pooled normal human urine, freeze dried, containing 0.075 mg. codeine. 0.075 mg. methadone. 0.075 mg. meperidine and 0.075 mg. morphine, reconstituted with 25 ml. of distilled water. Available commercially from Lederle Diagnostics. American Cyanamid Company. Pearl River. New York.
- One plate was sprayed with iodoplatinate reagent (A) and another plate with (B).
- a dry visualization reagent impregnated paper strip for use in preparing a spray reagent solution standard for visualization of a drug in urine in thin-layer chromatography which comprises a paper strip containing impregnated therein the dried residue of a completely impregnated volume of liquid solvent solution containing a predetermined amount of visualization reagent capable of elution into a spray reagent solvent and of chemically reacting with a drug in said urine to produce a characteristic color for said drug on a chromatogram.
- a method for preparing a dry visualization reagent impregnated strip for use in preparing a spray reagent solution standard for visualization of a drug in urine in thin-layer chromatography which comprises dipping an unimpregnated paper strip into a completely impregnated volume of a liquid solvent solution of a predetermined amount of visualization reagent capable of elution into a spray reagent solvent and of chemically reacting with a drug in said urine to produce a characteristic colored spot for said drug on a chromatogram and drying said strip.
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Abstract
Description
Claims (6)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
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US428565A US3912655A (en) | 1973-12-26 | 1973-12-26 | Reagent impregnated paper strips for use in preparing TLC spray reagents |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US428565A US3912655A (en) | 1973-12-26 | 1973-12-26 | Reagent impregnated paper strips for use in preparing TLC spray reagents |
Publications (1)
Publication Number | Publication Date |
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US3912655A true US3912655A (en) | 1975-10-14 |
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US428565A Expired - Lifetime US3912655A (en) | 1973-12-26 | 1973-12-26 | Reagent impregnated paper strips for use in preparing TLC spray reagents |
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Cited By (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4110078A (en) * | 1976-11-26 | 1978-08-29 | Zelonis Paul Thomas | Binary reagent system for detecting drugs of abuse |
US4193766A (en) * | 1978-11-13 | 1980-03-18 | Miles Laboratories, Inc. | Device and method for preparation of a control solution for ketone determination |
US4274833A (en) * | 1979-10-18 | 1981-06-23 | Pickering Michael V | Ninhydrin reagent for use in amine and amino acid analyses |
US4320086A (en) * | 1981-01-05 | 1982-03-16 | Andre Reiss | Paper device for rapid detection of cocaine |
FR2537724A1 (en) * | 1982-12-08 | 1984-06-15 | Medical Diagnostics Inc | Drug abuse test indicator and device and process for its manufacture |
US4529704A (en) * | 1979-11-05 | 1985-07-16 | Miles Laboratories, Inc. | Device and method for preparation of a control solution for ketone determination. |
WO1986003009A1 (en) * | 1984-11-16 | 1986-05-22 | Med-Tech Products, Inc. | Process for the preparation of a bibulous reactive medium |
US4605629A (en) * | 1980-12-23 | 1986-08-12 | Boehringer Mannheim Gmbh | Method of eluting reagent from reagent strips for chemical analyses and reagent strip therefor |
WO1987003961A1 (en) * | 1985-12-18 | 1987-07-02 | Keystone Diagnostics, Inc. | Drug abuse test paper and methods |
US4844866A (en) * | 1984-11-13 | 1989-07-04 | Matrix Technologies, Inc. | Carrier for detecting drug abuse compounds |
EP0336827A1 (en) * | 1988-04-08 | 1989-10-11 | Marie France Le Fel | Method for the detection of narcotic traces and products for use in this method |
US4992296A (en) * | 1987-05-26 | 1991-02-12 | Gibson Jacob J | Abuse-type drug test papers and methods of making and using same |
US5606116A (en) * | 1994-10-29 | 1997-02-25 | Kabushiki Kaisha Nippon Gene | Chromatogenous testing system for urinalysis |
US5710372A (en) * | 1996-12-17 | 1998-01-20 | Cincinnati Milacron Inc. | Method of analysis for aqueous fluids |
US5896692A (en) * | 1996-02-29 | 1999-04-27 | Collora; Samuel C. | Freeze dried scent lures |
WO2013188421A1 (en) * | 2012-06-11 | 2013-12-19 | The Curators Of The University Of Missouri | Cyclic germanium compounds and applications thereof |
US10369582B2 (en) | 2015-04-30 | 2019-08-06 | Emissol Llc | System and method for spray visualization |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
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US2229155A (en) * | 1938-04-23 | 1941-01-21 | Wenker Henry | Indicator paper |
US3046779A (en) * | 1961-04-25 | 1962-07-31 | Charles H Coleman | Solvent-front indicator for paper chromatograms |
US3138544A (en) * | 1961-05-03 | 1964-06-23 | British Drug Houses Canada Ltd | Microbial sensitivity testing device |
US3464796A (en) * | 1966-10-31 | 1969-09-02 | Saul Friedlander | Method of determining area |
US3586483A (en) * | 1968-10-14 | 1971-06-22 | American Cyanamid Co | Method for detecting the presence of tetracycline antibiotics |
US3590006A (en) * | 1967-09-13 | 1971-06-29 | Allied Chem | Aerosol compositions useful in preparation of thin-layer chromatography plates |
US3625652A (en) * | 1970-03-17 | 1971-12-07 | Marquette School Of Medicine I | Analysis of narcotics and amphetamines |
US3761227A (en) * | 1972-04-24 | 1973-09-25 | Conrad Labor Inc | Method for detection of opium derivatives in urine |
US3832134A (en) * | 1971-10-04 | 1974-08-27 | D Sohn | Tlc method and device for detecting the presence of target substances in unknown solutions |
-
1973
- 1973-12-26 US US428565A patent/US3912655A/en not_active Expired - Lifetime
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US2229155A (en) * | 1938-04-23 | 1941-01-21 | Wenker Henry | Indicator paper |
US3046779A (en) * | 1961-04-25 | 1962-07-31 | Charles H Coleman | Solvent-front indicator for paper chromatograms |
US3138544A (en) * | 1961-05-03 | 1964-06-23 | British Drug Houses Canada Ltd | Microbial sensitivity testing device |
US3464796A (en) * | 1966-10-31 | 1969-09-02 | Saul Friedlander | Method of determining area |
US3590006A (en) * | 1967-09-13 | 1971-06-29 | Allied Chem | Aerosol compositions useful in preparation of thin-layer chromatography plates |
US3586483A (en) * | 1968-10-14 | 1971-06-22 | American Cyanamid Co | Method for detecting the presence of tetracycline antibiotics |
US3625652A (en) * | 1970-03-17 | 1971-12-07 | Marquette School Of Medicine I | Analysis of narcotics and amphetamines |
US3832134A (en) * | 1971-10-04 | 1974-08-27 | D Sohn | Tlc method and device for detecting the presence of target substances in unknown solutions |
US3761227A (en) * | 1972-04-24 | 1973-09-25 | Conrad Labor Inc | Method for detection of opium derivatives in urine |
Cited By (22)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4110078A (en) * | 1976-11-26 | 1978-08-29 | Zelonis Paul Thomas | Binary reagent system for detecting drugs of abuse |
US4193766A (en) * | 1978-11-13 | 1980-03-18 | Miles Laboratories, Inc. | Device and method for preparation of a control solution for ketone determination |
US4274833A (en) * | 1979-10-18 | 1981-06-23 | Pickering Michael V | Ninhydrin reagent for use in amine and amino acid analyses |
US4529704A (en) * | 1979-11-05 | 1985-07-16 | Miles Laboratories, Inc. | Device and method for preparation of a control solution for ketone determination. |
US4605629A (en) * | 1980-12-23 | 1986-08-12 | Boehringer Mannheim Gmbh | Method of eluting reagent from reagent strips for chemical analyses and reagent strip therefor |
US4320086A (en) * | 1981-01-05 | 1982-03-16 | Andre Reiss | Paper device for rapid detection of cocaine |
FR2537724A1 (en) * | 1982-12-08 | 1984-06-15 | Medical Diagnostics Inc | Drug abuse test indicator and device and process for its manufacture |
WO1984002397A1 (en) * | 1982-12-08 | 1984-06-21 | Medical Diagnostics Inc | Drug abuse test indicator device and method of making same |
US4844866A (en) * | 1984-11-13 | 1989-07-04 | Matrix Technologies, Inc. | Carrier for detecting drug abuse compounds |
WO1986003009A1 (en) * | 1984-11-16 | 1986-05-22 | Med-Tech Products, Inc. | Process for the preparation of a bibulous reactive medium |
US4752448A (en) * | 1985-12-18 | 1988-06-21 | Keystone Diagnostics, Inc. | Drug abuse test paper |
WO1987003961A1 (en) * | 1985-12-18 | 1987-07-02 | Keystone Diagnostics, Inc. | Drug abuse test paper and methods |
US4992296A (en) * | 1987-05-26 | 1991-02-12 | Gibson Jacob J | Abuse-type drug test papers and methods of making and using same |
EP0336827A1 (en) * | 1988-04-08 | 1989-10-11 | Marie France Le Fel | Method for the detection of narcotic traces and products for use in this method |
FR2629915A1 (en) * | 1988-04-08 | 1989-10-13 | Lefel Marie France | METHOD FOR DETECTING TRACES OF NARCOTICS AND PRODUCTS FOR CARRYING OUT SAID METHOD |
US5606116A (en) * | 1994-10-29 | 1997-02-25 | Kabushiki Kaisha Nippon Gene | Chromatogenous testing system for urinalysis |
US5896692A (en) * | 1996-02-29 | 1999-04-27 | Collora; Samuel C. | Freeze dried scent lures |
US5710372A (en) * | 1996-12-17 | 1998-01-20 | Cincinnati Milacron Inc. | Method of analysis for aqueous fluids |
WO2013188421A1 (en) * | 2012-06-11 | 2013-12-19 | The Curators Of The University Of Missouri | Cyclic germanium compounds and applications thereof |
US10294252B2 (en) | 2012-06-11 | 2019-05-21 | The Curators Of The University Of Missouri | Cyclic germanium compounds and applications thereof |
US11352379B2 (en) | 2012-06-11 | 2022-06-07 | The Curators Of The University Of Missouri | Cyclic germanium compounds and applications thereof |
US10369582B2 (en) | 2015-04-30 | 2019-08-06 | Emissol Llc | System and method for spray visualization |
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