DE2910102A1 - Enhanced sensitivity of thin layer chromatography fluorimetric measure - using mirror glass sepn. layer substrate - Google Patents

Enhanced sensitivity of thin layer chromatography fluorimetric measure - using mirror glass sepn. layer substrate

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Publication number
DE2910102A1
DE2910102A1 DE19792910102 DE2910102A DE2910102A1 DE 2910102 A1 DE2910102 A1 DE 2910102A1 DE 19792910102 DE19792910102 DE 19792910102 DE 2910102 A DE2910102 A DE 2910102A DE 2910102 A1 DE2910102 A1 DE 2910102A1
Authority
DE
Germany
Prior art keywords
thin layer
sepn
mirror glass
fluorimetric
sensitivity
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.)
Withdrawn
Application number
DE19792910102
Other languages
German (de)
Inventor
Theodor Prof Dr Eckert
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to DE19792910102 priority Critical patent/DE2910102A1/en
Publication of DE2910102A1 publication Critical patent/DE2910102A1/en
Withdrawn legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
    • G01N30/92—Construction of the plate
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2201/00—Features of devices classified in G01N21/00
    • G01N2201/06—Illumination; Optics
    • G01N2201/061—Sources

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  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

Sensitivity of fluorimetric measurements of substances following thin layer chromatographic sepn., esp. High Performance Thin Layer Chromatography, involves the use of mirror glass instead of the conventional ordinary glass with mirrored surfaced as a substrate for the sepg. thin layer material. The thin sepn. layer is attached to the mirror glass. The sensitivity increase is partly dependent on reflection back of fluorescent light not radiated downwards tending to increase capture efficiency. Excitation light failing to produce fluorescence is reflected and passed through the thin layer plate again. In this way the amt. of fluorescence stimulated is additively increased.

Description

Verfahren zur Erhöhung der Empfindlichkeit bei fluorimetrischenMethod of increasing the sensitivity in fluorimetric

Messungen von Substanzen nach dünnschichtchromatographischer Trennung Bei dünnschichtchromatographischer Trennung von Substanzgemischen,insbesondere bei der sogenannten High Performance Thin Layer Chromatography (HPTLC),kann eine direkte quantitative Auswertung der getrennten Substanzen auf fluorimetrischer Basis auf der Dünnschichtplatte erfolgen.Dabei wird die Intensität des zurückgestrahlten Fluoreszenzlichtes ermittelt.Measurements of substances after separation by thin layer chromatography With thin-layer chromatographic separation of substance mixtures, especially with the so-called High Performance Thin Layer Chromatography (HPTLC), a direct quantitative evaluation of the separated substances on a fluorometric basis The intensity of the reflected fluorescent light determined.

Als Trennmedium für solche Analysen werden im allgemeinen Platten aus normalem Glas verwendet'die in geeigneter Weise mit Trennmedien beschichtet sind.Im Anschluß daran erfolgt die Auftrennung der Gemische durch flüssige Laufmittel,wobei die verschiedensten Verfahren üblich sind.Die getrennten Stoffe sind dann an verschiedenen Punkten der beschichteten Platte lokalisiert und können direkt durch Messung der Intensität ihres Fluoreszenzlichtes bestimmt werden.Dle Empfindlichkeit dieser Methodik ist jedoch in mannigfaltiger Weise begrenzt.Plates are generally used as the separation medium for such analyzes made of normal glass used'that suitably coated with separating media The mixtures are then separated by means of liquid mobile solvents, whereby The most diverse procedures are common. The separated substances are then at different Points of the coated plate are localized and can be viewed directly by measuring the The intensity of their fluorescent light can be determined. The sensitivity of this methodology however, it is limited in a number of ways.

Das im folgenden beschriebene Verfahren ermöglicht es,die Erfassungsempfindlichkeit der auf die beschriebene Weise getrennten Substanzen,soweit sie fluorimetrisch erfaßt werden können, zu erhöhen.Das Verfahren besteht erfindungsgemäß darin,anstelle des sonst für die trennende Dünnschicht verwendeten normalen Glases verspiegelte Flächen,z.B. Spiegelglas,-zu verwenden,auf das die trennende Dünnschicht fest aufgezogen wurde.Die empfindlichkeitssteigernde Wirkung der Erfindung ist in mehreren Faktoren begründet.Unter anderem wird Fluoreszenzlichtldas nach unten abstrahltlan der Spiegelfläche rückreflektiert und mit erfaßt. Anregungs licht, das die Dünnschichtplatte durchdrungen hat,ohne eine Fluoreszenz anzuregen,wird reflektiert und durchdringt erneut die fluoreszierende Schicht,wobei es zu einer additiven Verstärkung des zu messenden Fluoreszenzlichtes kommt.The procedure described below enables the detection sensitivity the substances separated in the manner described, insofar as they are determined fluorimetrically The method according to the invention consists in instead of of Normal glass mirrored surfaces otherwise used for the separating thin layer, e.g. Mirror glass to be used, onto which the separating thin layer has been firmly attached The sensitivity-increasing effect of the invention is based on several factors among other things, fluorescent light that is radiated downwards is reflected back on the mirror surface and captured with. Excitation light that has penetrated the thin-layer plate without stimulating a fluorescence is reflected and penetrates the fluorescent one again Layer, whereby there is an additive amplification of the fluorescent light to be measured comes.

Claims (1)

PATENTANSPRUCH Erhöhung der Empfindlichkeit zur Erfassung fluoreszierender Substanzen auf mit verschiedenen Trennmitteln beschichteten Dünnschichtplatten dadurch gekennzeichnet,daß als Unterlage für die trennende Dünnschicht eine verspiegelte Fläche verwendet wird (z.B. Spiegelglas).PATENT CLAIM Increasing the sensitivity for the detection of fluorescent substances on thin-layer plates coated with various separating agents, characterized in that a mirrored surface is used as a base for the separating thin layer (eg mirror glass).
DE19792910102 1979-03-15 1979-03-15 Enhanced sensitivity of thin layer chromatography fluorimetric measure - using mirror glass sepn. layer substrate Withdrawn DE2910102A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
DE19792910102 DE2910102A1 (en) 1979-03-15 1979-03-15 Enhanced sensitivity of thin layer chromatography fluorimetric measure - using mirror glass sepn. layer substrate

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
DE19792910102 DE2910102A1 (en) 1979-03-15 1979-03-15 Enhanced sensitivity of thin layer chromatography fluorimetric measure - using mirror glass sepn. layer substrate

Publications (1)

Publication Number Publication Date
DE2910102A1 true DE2910102A1 (en) 1980-09-25

Family

ID=6065413

Family Applications (1)

Application Number Title Priority Date Filing Date
DE19792910102 Withdrawn DE2910102A1 (en) 1979-03-15 1979-03-15 Enhanced sensitivity of thin layer chromatography fluorimetric measure - using mirror glass sepn. layer substrate

Country Status (1)

Country Link
DE (1) DE2910102A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2502051A1 (en) * 2009-11-20 2012-09-26 GE Healthcare Bio-Sciences AB System and method for increased fluorescence detection

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2502051A1 (en) * 2009-11-20 2012-09-26 GE Healthcare Bio-Sciences AB System and method for increased fluorescence detection
EP2502051A4 (en) * 2009-11-20 2013-07-03 Ge Healthcare Bio Sciences Ab System and method for increased fluorescence detection

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