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US3746869A - Method of photometrically plotting light scattering objects - Google Patents

Method of photometrically plotting light scattering objects Download PDF

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Publication number
US3746869A
US3746869A US00184416A US3746869DA US3746869A US 3746869 A US3746869 A US 3746869A US 00184416 A US00184416 A US 00184416A US 3746869D A US3746869D A US 3746869DA US 3746869 A US3746869 A US 3746869A
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United States
Prior art keywords
light
support
photometrically
detector
plotting
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Expired - Lifetime
Application number
US00184416A
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English (en)
Inventor
S Lindstedt
L Treiber
P Stollnberger
W Tausch
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Carl Zeiss AG
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Carl Zeiss AG
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47Scattering, i.e. diffuse reflection
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/255Details, e.g. use of specially adapted sources, lighting or optical systems
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47Scattering, i.e. diffuse reflection
    • G01N21/4738Diffuse reflection, e.g. also for testing fluids, fibrous materials
    • G01N21/474Details of optical heads therefor, e.g. using optical fibres
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/59Transmissivity
    • G01N21/5907Densitometers
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/90Plate chromatography, e.g. thin layer or paper chromatography
    • G01N30/95Detectors specially adapted therefor; Signal analysis

Definitions

  • Thin layer-chromatography is a modern analytical method of separating mixtures of organic and inorganic substances.
  • the solution of the substance mixture which is to separate is applied as a spot or as a band at a specific position on the separating layer.
  • the plate is placed in a chamber containing a suitable fluid medium. in the ensuing development, the fluid medium travels across the separating layer, as by capillary. forces, thereby separating the substance mixture.
  • the different substances of the mixture are found at different positions of the separating layer, at which point it is necessary to evaluate the developed thin layer-chromatogram qualitatively and quantitatively.
  • the light-absorption measurement technique is generally applicable in the case of objects which cause no, or only a small amount of, light scattering, in which case a transmission measurement is made.
  • a transmission measurement is made.
  • it is necessary prior to a transmission measurement to make the object transparent by means of a liquid of a suitable index refraction. Due to the resulting elution effects, such a method of plotting thin layer-chromatograms is hardly recommendable.
  • the chromatogram is scanned by a monochromatic light beam in the direction of separation, and the intensity of the diffusely reflected measuring radiation is measured with a photoelectric detector and recorder, for instance as a function of position, by a suitable recorder. lf lightabsorbing substance spots are found on the separating layer, the reflected radiation will be attenuated in those wavelength ranges in which absorption bands would occur for transmission measurements of corresponding solutions. The intensity of the diffusely reflected measuring radiation is therefore indicative of the absorption caused by the substance.
  • the absorption-position curve obtained by the scanning can be evaluated quantitatively by means of the Kubelka-Munk-function.
  • the effect being measured is always degraded by a superimposed lightscattering function, which varies as a function of position on the separating layer; in the specific case of small substance volumes, this superimposed function becomes very detrimentally noticeable and deteriorates the limit of detection.
  • This position-dependent function is unspecific relative to the light-scattering substance and may result in a highly irregular base line of the curve being evaluated, making it difficult if not impossible in some cases to decide definitely whether one is observing an effect to be measured or an interference effect.
  • the method according to the invention is characterized by making simultaneous measurements of the diffusely reflected and of the transmitted radiation, for each plotted position of the object, the two signals thus obtained being means of a recorder.
  • the novel method has the advantage that the making of the measurement inherently and automatically eliminates scattering effects related to position of the separating layer and unspecific to the substance of the light scattering, while, at the same time the actual measuring signal specific to the substance is amplified.
  • the device for carrying out the novel method is advantageously designed so that the measuring radiation impinges vertically on the surface of the object being plotted, that a first photoelectric detector is arranged above this surface and a second one below the object, and that both detectors are connected with an arrangement for adding the signals.
  • This arrangement is preferably in connection with a recorder recording the resultant absorption-position curve.
  • FIGS. 1 to 6 of the accompanying drawings in which:
  • FIG. 1 is an absorption-position curve of a thin layerchromatogram, recorded by measuring the reflected radiation as well as the associated base line;
  • FIG. 2 is the absorption position curve of the same thin layer-chromatogram, recorded by measuring the transmitted radiation as well as the associated base line; of the invention, together with the associated base line; for carrying out the method of the invention;
  • FIG. is a schematic diagram showing one embodiment of a first device, in which photometer balls are provided for the integral detection of the radiation;
  • FIGS. 6 and 7 are schematic perspective diagrams showing further detection devices, in which fiberoptical equipment is provided for the supply of the measuring light as well as for the detection of the reflected and the transmitted radiation.
  • reference numeral 1 designates an absorption-position curve obtained by recording reflected radiation, in the plotting of a thin layer-chromatogram. As can be seen, the effect being measured has superimposed thereon a light-scattering function depending upon local position on the separating layer.
  • the base line associated with curve 1 is referenced 2, and the desired data is represented by the difference between photometric curves, suggested by the crosshatched area.
  • the base line is seen to be irregular, and the sufficiently accurate determination of the hatched area is very difficult when plotting.
  • a device as schematically illustrated in FIG. 4 may be used to determine the absorption-position curve.
  • Light from a source 3 is monochromatized in the monochromator 4 and, via a reflecting mirror 5, impinges upon a thin layer-chromatogram plate referenced 6. Measuring light impinges upon the separating layer of the chromatogram 6 at an angle of 0 to the surface normal.
  • Diffusely reflected radiation is measured by means of a photoelectric detector 7 while a detector 8 simultaneously measures transmitted radiation.
  • the signal generated by the detector 7 is amplified by means 9 and is then supplied to an adding stage 1 1. In like manner, the latter also receives the signal output of detector 8 after amplification at means 10.
  • the two amplifiers 9 10 are preferably independently variable, as suggested by manually adjustable elements 9' 10', so that the amplitudes of signals unspecific to the substance can be caused to assume the same level.
  • the additive signal provided by means 11 is supplied to a suitable recorder l2.
  • the arrow symbol 13 will be understood to suggest means for traversing the chromatogram plate 6 and the described optical system with respect to each other, and the synchronizing connection 13' to the recorder 12 assures proper correlation of the summation signal with traverse position.
  • the absorption-position curve recorded by the recorder 12 is illustrated at 14 in FIG. 3.
  • the base line associated with this curve is also illustrated in FIG. 3 and is referenced 14'.
  • the desired data is the difference between curves 14 14', but since the base line curve 14' is so nearly flat, the curve 14 alone can be relied upon to yield the desired data.
  • the base line 14' has now been flattened to such extent that variations due to irregularities of the separating layer are only in the order of magnitude of electronic instability of the whole system (i.e. system noise).
  • evaluation of curve I4 is relatively simple and can be carried out with great accuracy.
  • the method of the invention will be seen to extend the limit of detection, inasmuch as deflections of the curve 14 (insofar as they are taken into consideration for an evaluation) are attributable only to effects specific to the substance.
  • a photometer ball 15 is arranged above the chromatogram plate 6; photometer ball 15 has an opening 16 for passage of the measuring radiation 17 therethrough. Radiation diffusely reflected by the separating layer of the plate 6 is detected by photoelectric means 18, in accordance with the various reflections within the photometer ball 15, as suggested by,dashed lines. Below plate 6, another photometer ball 19 including a photoelectric detector 20 is arranged to respond to transmitted light, the radiation passing through the separating layer of the chromatogram plate 6 reaching the detector 20 after several reflections within the photometer ball 19, again as suggested by dashed lines. In the embodiment illustrated in FlG. 5, a substantially integral detection of both the reflected and the transmitted radiation is achieved, thereby further enhancing the 'accuracy of measurement, as compared with the photocell arrangement in FIG. 4.
  • reference numeral 6 again designates a chromatogram plate.
  • the measuring radiation 17 is directed via a bundle of glass fibres 21 to the local area or position of the plate 6 being plotted.
  • the bundle 21 is divided at this position, and the partial bundle 21 directes the diffusely reflected light to a photocell 22.
  • another bundle of glass fibers 23 is arranged to receive the transmitted radiation, dirdcting the same to a second photoeldctric detector 24.
  • the method of photometrically evaluating lightabsorbing zones resulting upon the separation of mixtures of substances in thin layers of light-dispersing material which comprises irradiating light upon one localized area of the material, performing a traverse of said area over one such zone, simultaneously photometrically and additively measuring diffusely reflected light and transmitted light at said area in the course of the traverse, and recording the additive value as a function of traverse position.
  • the method of photometrically plotting light scattered by a light-scattering object which comprises irradiating light upon one surface of the object, simultaneously photometrically measuring diffusely reflected light and transmitted light for a given location on the object, and adding the simultaneously measured values; the given location being one of a succession involved in a transverse of the object, the added values being plotted as a function of traverse position, and the baseline amplitudes of the two measuring signals being caused to assume the same level, prior to adding the simultaneously measured values.
  • Apparatus for photometrically plotting light scattered by a light scattering object comprising a transpardnt support for the object, means for lightirradiating a limited surface area of the object normal to the support; photoelectric-detector means comprising a first detector on one side of the support for responding to diffusely reflected light from said area of the object, and a second detector on the other side of the support for responding simultaneously to directly transmitted light at said local area; means adding the outputs of said detectors, means for transversing said detector means and said support with respect to each other, and recorder means synchronized with traverse displacement and responsive to the added outputs of said detectors.
  • detectors are photometer balls disposed above and below said support, in vertically aligned relation, each photometer ball including a photoelectric cell.
  • said detector means includes fiber-optical means disposed on said one side of said support for conducting reflected light to said first detector.
  • said detector means includes fiber-optical means disposed on said other side of said support for conducting transmitted light to said second detector.
  • Apparatus for photometrically plotting light scattered by a lightscattering object comprising a transparent support for the object, means for lightirradiating the object normal to the support, and detection means comprising fiber-optical means having a first fiber-optical bundle with ends adjacent one side of the support for responding to diffusely reflected light from a local area of the object, said fiber-optical means having a second fiber-optical bundle with ends adjacent the other side. of the support for responding to directly transmitted light at said local area; and means including photoelectric means responsive to the sum of the light components transmitted by said first and second bundles.
  • Apparatus according to claim 14 in which said light-irradiating means includes a further fiber-optical bundle with ends on said one side of said support and terminating normal thereto at said local area.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (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)
  • Spectroscopy & Molecular Physics (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
US00184416A 1970-09-30 1971-09-28 Method of photometrically plotting light scattering objects Expired - Lifetime US3746869A (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
DE2047952A DE2047952C3 (de) 1970-09-30 1970-09-30 Verfahren zur photometrischen Auswertung der sich bei der Auftrennung von Substanz gemischen in dünnen Schichten aus licht streuendem Material ergebenden Zonen

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GB (1) GB1328734A (de)

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3994587A (en) * 1974-08-21 1976-11-30 Shimadzu Seisakusho Ltd. Densitometer
US4012144A (en) * 1975-08-07 1977-03-15 Varian Associates Spectrosorptance measuring system and method
FR2360072A1 (fr) * 1976-07-30 1978-02-24 Industrial Nucleonics Corp Procede et appareil de mesure d'une propriete d'un materiau, notamment sa teneur en humidite
US4120582A (en) * 1976-10-27 1978-10-17 Donnelly Mirrors, Inc. Light reflectivity and transmission testing apparatus and method
FR2409507A1 (fr) * 1977-11-17 1979-06-15 Lommel Hermann Procede de determination quantitative d'elements troubles provenant en particulier de reactions d'immunite
US4171909A (en) * 1977-03-25 1979-10-23 Miles Laboratories, Inc. Apparatus for measuring light intensities
FR2524162A1 (fr) * 1982-03-23 1983-09-30 Canon Kk Appareil et procede de controle de negatifs et dispositif de cadrage de masques
JPS59206747A (ja) * 1983-03-21 1984-11-22 ドナルド・ダブリユ−・バイヤ−ズ 被覆検知と表面評価のための方法及び装置
US4492462A (en) * 1980-02-16 1985-01-08 Wilhelm Pross Photometric method for determining courses of reactions
US4752696A (en) * 1986-05-19 1988-06-21 Fuji Photo Film Co., Ltd. Method of inspecting floppy disk casing
US4810872A (en) * 1986-07-26 1989-03-07 Hitachi, Ltd. Optical property measuring device
US4937637A (en) * 1989-02-10 1990-06-26 Kollmorgen Corporation Dual reading head transmission/reflection densitometer
WO1992000514A1 (en) * 1990-06-29 1992-01-09 Tma Technologies, Inc. Modular scatterometer with interchangeable scanning heads
US5717216A (en) * 1996-10-16 1998-02-10 Reynolds Metals Company Thickness gauging using ultraviolet light absorption
US6485687B1 (en) 1998-03-14 2002-11-26 Bernd Spangenberg Thin-layer chromatography apparatus
US8970830B2 (en) 2011-06-09 2015-03-03 Carl Zeiss Microscopy Gmbh Measuring method and device for determining transmission and/or reflection properties
US20160377535A1 (en) * 2015-06-23 2016-12-29 Empire Technology Development Llc Real-time monitoring of material composition for quality control

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2757196C3 (de) * 1977-12-22 1981-11-26 Vladimir Dr.-Ing. 5100 Aachen Blazek Photometrische Anordnung
US4199260A (en) * 1978-08-21 1980-04-22 Technicon Instruments Corporation Apparatus and method for determining the concentration in a sample
DE3213533A1 (de) * 1982-04-10 1983-10-20 Bruker Analytische Meßtechnik GmbH, 7512 Rheinstetten Infrarot-spektrometer

Citations (10)

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US2632855A (en) * 1947-10-07 1953-03-24 Westinghouse Electric Corp Photoelectric speed control
US2718597A (en) * 1951-11-01 1955-09-20 Exxon Research Engineering Co Infrared analysis apparatus
US2834247A (en) * 1955-01-26 1958-05-13 Beckman Instruments Inc Optical density analyzing apparatus
US2930898A (en) * 1956-09-11 1960-03-29 W D Engineering Co Ltd Cathode ray tube apparatus for the inspection of articles
US3320428A (en) * 1963-05-06 1967-05-16 British Petroleum Co Photosensitive apparatus for detecting solid particles suspended in liquid
US3504978A (en) * 1965-02-04 1970-04-07 Shimadzu Corp Plural beam spectrophotometer with a diffusion plate between each cell and detector
US3504983A (en) * 1966-05-31 1970-04-07 Nasa Ellipsoidal mirror reflectometer including means for averaging the radiation reflected from the sample
US3566083A (en) * 1967-10-16 1971-02-23 Measurement Research Center In Sensor for punches and marks
US3567328A (en) * 1966-06-02 1971-03-02 Vernon T Riley Specimen transport mount for spectrophotometer
US3618061A (en) * 1969-04-30 1971-11-02 Eaton Yale & Towne Monitoring apparatus for monitoring the density of a material carried by a fluid and the flow of the fluid

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2632855A (en) * 1947-10-07 1953-03-24 Westinghouse Electric Corp Photoelectric speed control
US2718597A (en) * 1951-11-01 1955-09-20 Exxon Research Engineering Co Infrared analysis apparatus
US2834247A (en) * 1955-01-26 1958-05-13 Beckman Instruments Inc Optical density analyzing apparatus
US2930898A (en) * 1956-09-11 1960-03-29 W D Engineering Co Ltd Cathode ray tube apparatus for the inspection of articles
US3320428A (en) * 1963-05-06 1967-05-16 British Petroleum Co Photosensitive apparatus for detecting solid particles suspended in liquid
US3504978A (en) * 1965-02-04 1970-04-07 Shimadzu Corp Plural beam spectrophotometer with a diffusion plate between each cell and detector
US3504983A (en) * 1966-05-31 1970-04-07 Nasa Ellipsoidal mirror reflectometer including means for averaging the radiation reflected from the sample
US3567328A (en) * 1966-06-02 1971-03-02 Vernon T Riley Specimen transport mount for spectrophotometer
US3566083A (en) * 1967-10-16 1971-02-23 Measurement Research Center In Sensor for punches and marks
US3618061A (en) * 1969-04-30 1971-11-02 Eaton Yale & Towne Monitoring apparatus for monitoring the density of a material carried by a fluid and the flow of the fluid

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3994587A (en) * 1974-08-21 1976-11-30 Shimadzu Seisakusho Ltd. Densitometer
US4012144A (en) * 1975-08-07 1977-03-15 Varian Associates Spectrosorptance measuring system and method
FR2360072A1 (fr) * 1976-07-30 1978-02-24 Industrial Nucleonics Corp Procede et appareil de mesure d'une propriete d'un materiau, notamment sa teneur en humidite
US4120582A (en) * 1976-10-27 1978-10-17 Donnelly Mirrors, Inc. Light reflectivity and transmission testing apparatus and method
US4171909A (en) * 1977-03-25 1979-10-23 Miles Laboratories, Inc. Apparatus for measuring light intensities
FR2409507A1 (fr) * 1977-11-17 1979-06-15 Lommel Hermann Procede de determination quantitative d'elements troubles provenant en particulier de reactions d'immunite
US4492462A (en) * 1980-02-16 1985-01-08 Wilhelm Pross Photometric method for determining courses of reactions
US4669885A (en) * 1982-03-23 1987-06-02 Canon Kabushiki Kaisha Apparatus for inspecting negatives
FR2524162A1 (fr) * 1982-03-23 1983-09-30 Canon Kk Appareil et procede de controle de negatifs et dispositif de cadrage de masques
JPS59206747A (ja) * 1983-03-21 1984-11-22 ドナルド・ダブリユ−・バイヤ−ズ 被覆検知と表面評価のための方法及び装置
EP0119630A3 (de) * 1983-03-21 1985-11-27 Donald W. Byers Verfahren und Einrichtung zur Feststellung einer Deckschicht und Abschätzung einer Oberfläche
US4752696A (en) * 1986-05-19 1988-06-21 Fuji Photo Film Co., Ltd. Method of inspecting floppy disk casing
US4810872A (en) * 1986-07-26 1989-03-07 Hitachi, Ltd. Optical property measuring device
US4937637A (en) * 1989-02-10 1990-06-26 Kollmorgen Corporation Dual reading head transmission/reflection densitometer
WO1992000514A1 (en) * 1990-06-29 1992-01-09 Tma Technologies, Inc. Modular scatterometer with interchangeable scanning heads
US5196906A (en) * 1990-06-29 1993-03-23 Tma Technologies, Inc. Modular scatterometer with interchangeable scanning heads
US5717216A (en) * 1996-10-16 1998-02-10 Reynolds Metals Company Thickness gauging using ultraviolet light absorption
US6485687B1 (en) 1998-03-14 2002-11-26 Bernd Spangenberg Thin-layer chromatography apparatus
US8970830B2 (en) 2011-06-09 2015-03-03 Carl Zeiss Microscopy Gmbh Measuring method and device for determining transmission and/or reflection properties
US20160377535A1 (en) * 2015-06-23 2016-12-29 Empire Technology Development Llc Real-time monitoring of material composition for quality control
US10197496B2 (en) * 2015-06-23 2019-02-05 Empire Technology Development Llc Real-time monitoring of material composition for quality control

Also Published As

Publication number Publication date
DE2047952B2 (de) 1973-03-22
DE2047952C3 (de) 1973-10-18
DE2047952A1 (de) 1972-04-06
GB1328734A (en) 1973-08-30

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