DE3507171A1 - Compact fluorimeter with optimum excitation geometry - Google Patents

Compact fluorimeter with optimum excitation geometry

Info

Publication number
DE3507171A1
DE3507171A1 DE19853507171 DE3507171A DE3507171A1 DE 3507171 A1 DE3507171 A1 DE 3507171A1 DE 19853507171 DE19853507171 DE 19853507171 DE 3507171 A DE3507171 A DE 3507171A DE 3507171 A1 DE3507171 A1 DE 3507171A1
Authority
DE
Germany
Prior art keywords
light source
excitation
focal axis
compact
fluorimeter
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.)
Ceased
Application number
DE19853507171
Other languages
German (de)
Inventor
Reinhard Dr. 5840 Schwerte Nießner
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 DE19853507171 priority Critical patent/DE3507171A1/en
Publication of DE3507171A1 publication Critical patent/DE3507171A1/en
Ceased legal-status Critical Current

Links

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
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • 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
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N2021/6463—Optics
    • G01N2021/6469—Cavity, e.g. ellipsoid
    • 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
    • G01N21/84—Systems specially adapted for particular applications
    • G01N21/85—Investigating moving fluids or granular solids
    • 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
    • G01N2201/069—Supply of sources
    • G01N2201/0696—Pulsed
    • 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
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/74—Optical detectors

Abstract

In the conventional measuring configuration of a fluorimetric analysis, only a small part of the radiation exciting the fluorescence is transmitted to the volume element containing the fluorescing substance. In the measuring configuration according to the invention, the volume element to be examined is placed in a focal axis of an elliptical bore. The rod-shaped excitation light source is located in the second focal axis and, by means of reflection, transmits a large proportion of the light to the volume element in the other focal axis. The measuring configuration is particularly suitable for use as a flow-through detector in combination with chromatographic separation processes (e.g. HPLC).

Description

Kompakt - Fluorimeter mit optimaler AnregungsReometrieCompact fluorimeter with optimal excitation geometry

Beschreibung : Die Erfindung betrifft eine kompakte Fluorimetrieanregungs- und meßanordnung mit optimaler Anregungsgeometrie, insbesonders in Kombination mit chemisch - analytischen Chromatographiesystemen.Description: The invention relates to a compact fluorimetry excitation and measuring arrangement with optimal excitation geometry, especially in combination with chemical - analytical chromatography systems.

Die vorgeschlagene Anregungsgeometrie soll die von der Anregungslichtquelle gelieferte Lichtintensität möglichst vollständig in das die zu analysierende Flüssigkeit enthaltende Volumenelement übertragen. Dadurch wird die für eine gegebene Lichtquelle höchstmögliche Nachweisstärke bei minimalster Wärmeverlustleistung der Anregungslichtquelle erreicht.The proposed excitation geometry should be that of the excitation light source delivered light intensity as completely as possible into the liquid to be analyzed Transfer containing volume element. This will be the most suitable for a given light source highest possible detection strength with minimal heat dissipation of the excitation light source achieved.

Seit vielen Jahren bedient sich die analytische Chemie der fluorimetrischen Analyse als nachweisstarkem Verfahren. Der Hauptvorteil der Fluorimetrie liegt dabei im linearen Zusammenhang zwischen der Anregungsintensität und der beobachteten Fluoreszenzemissionsstrahlung QF = Quantenausbeute der fluoreszierenden Substanz 10 = Intensität der Erregerstrahl-F = 2.3. E I0.c s ung = = molarer Extinktionskoeffizient s = Schichtdicke c = Konzentration der fluoreszierenden Substanz Für eine Erhöhung des Nachweisvermögens bei einem fluorimetrischen Verfahren bietet sich daher einmal die Erhöhung der Anregungsintensität an.For many years analytical chemistry has made use of fluorimetric Analysis as a method with strong evidence. The main advantage of fluorimetry lies in this in the linear relationship between the excitation intensity and the observed fluorescence emission radiation QF = quantum yield of the fluorescent substance 10 = intensity of the excitation beam F = 2.3. E I0.c s ung = = molar extinction coefficient s = layer thickness c = concentration of the fluorescent substance For an increase in the detection capability of a fluorimetric methods therefore offer an increase in the excitation intensity at.

Dies kann vorteilhaft z.B. durch eine Blitzlampe erreicht werden.This can advantageously be achieved, for example, with a flash lamp.

Wesentlicher ist jedoch die zum Gegenstand der vorliegenden Erfindung gemachte Anordnung, bei der das Anregungslicht möglichst vollständig auf den Gesamtinhalt einer Quarzküvette fokussiert wird. In üblichen Anordnungen mit geringem Öffnungswinkel der Lampe geht ein hoher Anteil an Anregungsintensität verloren oder wird nicht zur Anregung des gesamten Küvetteninhaltes benutzt. In der letzten Zeit werden parabolisch geformte Lampengehäuse zur Erzielung einer hohen Lichtintensität im Brennpunkt vorgeschlagen ( Lit. : M. Zander; " Fluorimetrie ", Springer - Verlag, Berlin, 1981, Seite 51 ). Für eine fluorimetrische Meßanordnung ist es jedoch vorteilhaft, wenn das zu analysierende Meßvolumen möglichst gleichmäßig von der anregenden Lichstrahlung durchsetzt wird.However, the subject matter of the present invention is more essential made arrangement in which the excitation light as completely as possible on the entire content a quartz cuvette is focused. In common arrangements with a small opening angle the lamp loses a high proportion of the excitation intensity or does not used to excite the entire contents of the cuvette. Lately they have become parabolic Shaped lamp housing proposed to achieve a high light intensity in the focal point (Lit.: M. Zander; "Fluorimetrie", Springer - Verlag, Berlin, 1981, page 51 ). For a fluorimetric measuring arrangement, however, it is advantageous if that is the case analyzing measuring volume as evenly as possible from the stimulating light radiation is enforced.

Dies ist deshalb von Bedeutung, da der Erhöhung der Anregungsintensität pro Volumenelement natürliche Grenzen gesetzt sind. Es ist daher günstiger das gesamte Küvettenvolumen gleichmäßig zu einer hohen Fluoreszenzdichte zu bringen, als z.B. durch Verwendung eines Lasers nur einen geringen Volumenelementanteil anzuregen und dafür Zersetzungs-, Quench-und Eigenfiltrationseffekte in Kauf nehmen zu müssen.This is important because of the increase in the excitation intensity there are natural limits per volume element. It is therefore cheaper the whole To bring cuvette volume uniformly to a high fluorescence density, as e.g. stimulate only a small proportion of the volume element by using a laser and having to accept decomposition, quenching and intrinsic filtration effects in return.

Optimal ist dagegen zum Gegenstand dieser Erfindung gemachte Meßanordnung ( siehe Abb. 1 ). In einer Brennachse 1 einer ellipsenförmigen Bohrung 2 befindet sich eine stabförmige Lichtquelle 3 ( z.B. der Firma Hamamatsu Typ L 937 - 004). Durch die allseitige Reflexion der Anregungsstrahlung an den Wänden der ellipsenförmigen Bohrung 2 wird das Abbild der anregenden Lichtquelle vollständig in die im zweiten Brennpunkt 4 der ellipsenförmigen Bohrung 2 befindlichen Quarzküvette 5 projeziert.On the other hand, the measurement arrangement which is the subject of this invention is optimal (see Fig. 1). Located in a focal axis 1 of an elliptical bore 2 a rod-shaped light source 3 (e.g. from Hamamatsu type L 937-004). Due to the all-round reflection of the excitation radiation on the walls of the elliptical Hole 2 is the image of the exciting light source completely in the second Focal point 4 of the elliptical bore 2 located quartz cuvette 5 is projected.

Als reflexionsfähiges Wandmaterial wird hochreines Aluminium verwendet.High-purity aluminum is used as the reflective wall material.

Die Fluoreszenzstrahlung wird senkrecht am Küvettenboden z.B. mittels einer Photodiode mit integriertem Operationsverstärker ( z. B. OSI 5 ) 6 registriert. Besonders geeignet für die Beobachtung der Fluoreszenzstrahlung von polyzyklischen Kohlenwasserstoffen ( PAH's ) ist eine Anregungswellenlänge von 254 nm. Das die Fluoreszenzemission registrierende Photoelement ist durch geeignete Maßnahmen gegenüber der Anregungswellenlänge insensitiv gemacht.The fluorescence radiation is perpendicular to the bottom of the cuvette, e.g. by means of a photodiode with an integrated operational amplifier (e.g. OSI 5) 6. Particularly suitable for observing the fluorescence radiation from polycyclic Hydrocarbons (PAH's) has an excitation wavelength of 254 nm. The die Fluorescence emission registering photo element is opposed by suitable measures made insensitive to the excitation wavelength.

Durch geeignete dünne stabförmige Anregungslichtquellen wird besonders vorteilhaft das Problem der optimalen Anregung bei der Detektion von aus Chromatographiesäulen fließenden, kleinen Volumina gelöst. Durch Anwendung einer kapillarartigen Durchflußküvette lassen sich auch ijl -Volumina ( z.B. aus HPLC - Systemen ) hochempfindlich ausmessen.Suitable thin rod-shaped excitation light sources are particularly advantageously the problem of optimal excitation when detecting from chromatography columns flowing, small volumes solved. By using a capillary-type flow cell ijl volumes (e.g. from HPLC systems) can also be measured with high sensitivity.

Ein weiterer wesentlicher Vorteil der zum Gegenstand der Erfindung gemachten Anordnung ist die im Gegensatz zu den bisherigen Fluoreszenzmeßgeräten wenig aufwendige Stromversorgung bei mindestens gleicher Anregungsintensität pro Volumenelement. Dies gilt besonders im Hinblick auf Hochdruck - Lampen. Bei Verwendung einer stabförmigen Blitzlampe läßt sich eine weitere Erhöhung der Nachweisstärke erzielen.Another major advantage of the subject of the invention The arrangement made is in contrast to the previous fluorescence measuring devices low-cost power supply with at least the same excitation intensity per Volume element. This is especially true with regard to high pressure lamps. Using a rod-shaped flash lamp can be used to further increase the detection strength achieve.

Durch die vorzugsweise angewandte Kopplung mit einem Chromatographie -System wird die höchste Selektivität und Nachweisstärke erreicht. Selbstverständlich kann anstatt des Vorganges der Fluoreszenzanregung auch der Vorgang der Fluoreszenzlöschung als Analysensignal beobachtet werden.Through the preferably used coupling with a chromatography System, the highest selectivity and detection strength is achieved. Of course Instead of the process of fluorescence excitation, the process of fluorescence quenching can also be used can be observed as an analysis signal.

Ohne Chromatographie - Trennschritt ist das Gerät zur schnellen Übersichtsprüfung auf das Vorhandensein fluoreszierender Stoffe im Feld geeignet. Für die Prüfung auf polyzyklische Aromaten wird z.B. Erde trocken gepresst und mit trockenem Kaliumchlorid zur Wasseraufnahme verrieben. Danach wird das Gemisch mit n - Heptan extrahiert und das Lösungsmittel von der Suspension mittels einer Membranfilter - Spritze abgetrennt und die im Eluat befindlichen PAH's zur Fluoreszenz bei 254 nm angeregt.Without a chromatography separation step, the device can be used for a quick overview check for the presence of fluorescent substances in the field suitable. For testing for polycyclic aromatics, e.g. soil is pressed dry and with triturated with dry potassium chloride to absorb water. Then the mixture is with n - Heptane is extracted and the solvent is extracted from the suspension using a membrane filter - Syringe separated and the PAHs in the eluate to fluorescence at 254 nm excited.

Claims (4)

Kompakt - Fluorimeter mit optimaler Anregungsgeometrie Patentansprüche 1. Fluorimetriemeßgerät, dadurch gekennzeichnet, daß sich die anregende, stabförmige Lichtquelle in einer Brennachse einer ellipsenförmigen Reflexionsanordnung befindet und die Küvette mit der zu vermessenden Flüssigkeit sich in der anderen Brennachse befindet und die Fluoreszenzstrahlung senkrecht zur Anregungsrichtung registriert wird.Compact fluorimeter with optimal excitation geometry Patent claims 1. Fluorimetriemeßgerät, characterized in that the stimulating, rod-shaped Light source is located in a focal axis of an elliptical reflection arrangement and the cuvette with the liquid to be measured is in the other focal axis and the fluorescence radiation is registered perpendicular to the direction of excitation will. 2. Fluorimetriemeßgerät nach Anspruch 1, dadurch gekennzeichnet, daß die zu vermessende Flüßigkeit kontinuierlich durch die Küvette gepumpt und die fluoreszierende Strahlung ebenfalls kontinuierlich registriert wird.2. Fluorimetry measuring device according to claim 1, characterized in that the liquid to be measured is continuously pumped through the cuvette and the fluorescent one Radiation is also registered continuously. 3. Fluorimetriemeßgerät nach Anspruch 1 und 2, dadurch gekennzeichnet, daß es als kontinuierlich registrierendes System mit einem Chromatographie - Trennsystem zur qualitativen und quantitativen Bestimmung fluoreszierender Substanzen kombiniert wird.3. Fluorimetry measuring device according to claim 1 and 2, characterized in that that it is a continuously registering system with a chromatography separation system combined for the qualitative and quantitative determination of fluorescent substances will. 4. Fluorimetriemeßgerät nach einem der Ansprüche 1 - 3, dadurch gekennzeichnet, daß als anregende Lichtquelle eine gepulste Lichtquelle verwendet wird.4. Fluorimetry measuring device according to one of claims 1 - 3, characterized in that that a pulsed light source is used as a stimulating light source.
DE19853507171 1985-03-01 1985-03-01 Compact fluorimeter with optimum excitation geometry Ceased DE3507171A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
DE19853507171 DE3507171A1 (en) 1985-03-01 1985-03-01 Compact fluorimeter with optimum excitation geometry

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
DE19853507171 DE3507171A1 (en) 1985-03-01 1985-03-01 Compact fluorimeter with optimum excitation geometry

Publications (1)

Publication Number Publication Date
DE3507171A1 true DE3507171A1 (en) 1986-09-04

Family

ID=6263845

Family Applications (1)

Application Number Title Priority Date Filing Date
DE19853507171 Ceased DE3507171A1 (en) 1985-03-01 1985-03-01 Compact fluorimeter with optimum excitation geometry

Country Status (1)

Country Link
DE (1) DE3507171A1 (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4861163A (en) * 1988-01-06 1989-08-29 Amoco Corporation Ellipsoidal cylinder fluorescence analyzer
DE9004657U1 (en) * 1990-04-25 1990-07-05 Hewlett-Packard Gmbh, 7030 Boeblingen, De

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1797547A1 (en) * 1966-08-02 1971-11-25 Instrumentation Specialties Co Device for measuring light absorption, in particular extinction

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1797547A1 (en) * 1966-08-02 1971-11-25 Instrumentation Specialties Co Device for measuring light absorption, in particular extinction

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
Analytical Chemistry, Vol. 43, No. 4, 1971, S. 32A - 38A *
Chemie-Ing.-Techn., Jg. 39, 1967, S. 1008-1016 *
Journal of Physics E: Scientific Instrum., Vol. 7, 1974, S. 556-560 *
Prospekt der Fa. Camag, 4132 Muttenz/Schweiz, Homburgerstr. 24: Turner Fluorometer, 8 Seiten, eingeg. am 15.12.78 *
W.Kleen u. R.Müller (Herausg.): Laser, Springer-Verlag Berlin- Heidelberg-New York 1969, S.119-133 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4861163A (en) * 1988-01-06 1989-08-29 Amoco Corporation Ellipsoidal cylinder fluorescence analyzer
DE9004657U1 (en) * 1990-04-25 1990-07-05 Hewlett-Packard Gmbh, 7030 Boeblingen, De

Similar Documents

Publication Publication Date Title
EP0148497B1 (en) Device for guiding and collecting light in photometry or the like
Parker et al. Some experiments with spectrofluorimeters and filter fluorimeters
Turner An Absolute Spectrofluorometer: A new instrument has been developed for simple determinations of absolute fluorescence spectra.
Parker et al. Correction of fluorescence spectra and measurement of fluorescence quantum efficiency
US3849654A (en) Fluorescence cuvette
Fery-Forgues et al. Are fluorescence quantum yields so tricky to measure? A demonstration using familiar stationery products
Herman Resonance energy transfer microscopy
Winefordner et al. Determination of Zinc, Cadmium, and Mercury by Atomic Fluorescence Flame Spectormetry.
CN1043685C (en) Methods and apparatus for isotopic analysis
US5926271A (en) Laser-induced fluorescence detector having a capillary detection cell and method for identifying trace compounds implemented by the same device
DE4021955A1 (en) LASER OPERATED DETECTING DEVICE
Perry et al. Detection of ultratrace levels of uranium in aqueous samples by laser-induced fluorescence spectrometry
JPS62129741A (en) Photoacoustic analysis method and apparatus
Wehry Molecular fluorescence, phosphorescence, and chemiluminescence spectrometry
Winefordner et al. Phosphorimetry as a Means of Chemical Analysis. The Analysis of Aspirin in Blood Serum and Plasma.
Wehry Molecular fluorescence and phosphorescence spectrometry
Wirth et al. Very high detectability in two-photon spectroscopy
DE3507171A1 (en) Compact fluorimeter with optimum excitation geometry
DE3200391A1 (en) OPTICAL DEVICE FOR A FLUORESCENT POLARIZATION DEVICE
McGlynn et al. Total luminescence of organic molecules of petrochemical interest: Naphthalene, phenanthrene and 1, 2, 4, 5-tetramethylbenzene
US4128336A (en) Spectroscopic apparatus and method
Scholler et al. A study of estrogen fluorescence using Ittrich's procedure
DE2246365A1 (en) Nitric oxide determn. in air or exhaust gases - by spectroscopic analysis of visible radiation transmitted through sample
US4402606A (en) Optogalvanic intracavity quantitative detector and method for its use
Lamola et al. Recommended standards for reporting photochemical data (Recommendations 1983)

Legal Events

Date Code Title Description
OP8 Request for examination as to paragraph 44 patent law
8131 Rejection