WO2007041999A1 - Dispositif et procede pour realiser la determination qualitative et/ou quantitative de substances a activite ir contenues dans des liquides - Google Patents

Dispositif et procede pour realiser la determination qualitative et/ou quantitative de substances a activite ir contenues dans des liquides Download PDF

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Publication number
WO2007041999A1
WO2007041999A1 PCT/DE2006/001771 DE2006001771W WO2007041999A1 WO 2007041999 A1 WO2007041999 A1 WO 2007041999A1 DE 2006001771 W DE2006001771 W DE 2006001771W WO 2007041999 A1 WO2007041999 A1 WO 2007041999A1
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WO
WIPO (PCT)
Prior art keywords
liquid
measuring
sample
atr body
measured
Prior art date
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PCT/DE2006/001771
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German (de)
English (en)
Inventor
Werner MÄNTELE
Oliver Klein
Gamze Hosafci
Gerhard Oremek
Hermann Von Lilienfeld-Toal
Original Assignee
Johann Wolfgang Goethe-Universität Frankfurt am Main
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Application filed by Johann Wolfgang Goethe-Universität Frankfurt am Main filed Critical Johann Wolfgang Goethe-Universität Frankfurt am Main
Publication of WO2007041999A1 publication Critical patent/WO2007041999A1/fr

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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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/251—Colorimeters; Construction thereof
    • G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5088—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above confining liquids at a location by surface tension, e.g. virtual wells on plates, wires
    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
    • G01N21/3577—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing liquids, e.g. polluted water
    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/55—Specular reflectivity
    • G01N21/552—Attenuated total reflection
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0803—Disc shape
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0848—Specific forms of parts of containers
    • B01L2300/0858—Side walls
    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
    • G01N2021/3595—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using FTIR
    • 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
    • G01N2201/12—Circuits of general importance; Signal processing
    • G01N2201/129—Using chemometrical methods

Definitions

  • the present invention relates to a device for the qualitative and / or quantitative determination of IR-active ingredients in liquids as well as a method for the qualitative and / or quantitative determination of IR-active ingredients in liquids.
  • DE 103 28 998 A1 discloses an IR-ATR-based device for analyzing very small amounts of sample, in which a metering device for quantities in the nl range and an ATR device in a single instrument are combined.
  • components of the blood such as glucose, cholesterol, triglycerides, albumin, total protein and urea can only be determined with this device if the sample is dried on the measurement surface before measuring the ATR-IR spectrum. The drying of the sample to be measured is regularly accompanied by a considerable non-linearity in the determination of the concentration.
  • ATR immersion probes for measuring whole blood are described in US 5,170,056. Measurement solutions can be measured remotely from the actual spectrometer via a light guide.
  • An optical mass flow rate analysis device is disclosed in US 6,469,311. A large number of liquid samples are determined, for example, in a microtiter plate in a contactless manner by means of optical analysis methods comprising the measurement of the absorption, the photoluminescence and the chemiluminescence.
  • WO 96/04544 A1 also relates to an automated method for the determination of blood constituents.
  • the liquid sample is introduced into a recording device in order to detect scattered light as well as fluorescence signals. In this way, information about the red and white blood cells in the blood is available.
  • the device presented in WO 98/04544 A1 essentially comprises a combination of a conventional hematology analyzer and a fluorescence cytometric analyzer.
  • WO 00/66269 A1 has an integrated sample processing system for preparing samples and making them available for analysis.
  • the device proposed in WO 00/66269 A1 has to include a liquid source, a pump connected thereto, an output unit and a line from the pump to the dispensing unit, the line remaining open between successive contactless liquid dispensers having a volume of less than 5 ⁇ l.
  • recourse is preferably made to an output tip of a hydrophobic material. Further details, for example in the direction of specific analysis methods, WO 00/66269 Al can not be removed.
  • the devices used to measure a large number of samples at a high throughput are still very expensive in terms of apparatus and are associated in their results with relatively large error tolerances, especially if only very small amounts of sample are available.
  • the present invention therefore an object of the invention to provide a device that is no longer subject to the disadvantages of the prior art and in particular, even at high sample throughput, from very small liquid sample volumes, the simultaneous determination of a variety of parameters or ingredients It is also easy to operate and to load and easy to clean, without the need for special methods or materials.
  • an apparatus for the quantitative and / or qualitative determination of IR-active ingredients of aqueous or non-aqueous liquids
  • a first ATR body in the form of a light guide comprising a measuring surface in contact with the liquid to be measured which is transparent or partially transparent to the measuring radiation and which has a refractive index which is greater than that of the adjacent to the measuring surface during the measurement liquid, at least one second ATR body, the at least one flat boundary surface, in particular at least two planar, in comprising substantially parallel boundary surfaces, one of which is the measuring surface in contact with the liquid to be measured, and which is transparent or partially transparent to the measuring radiation and which has a refractive index which is greater than that of the liquid adjacent to the measuring surface during the measurement at least one sample holder, comprising at least one receiving device for the liquid to be measured with a bearing surface for this liquid and an edge which is dimensioned to make contact with the measuring surface of the liquid first or second ATR body with the surface of the liquid to be measured present in the recording device,
  • the measuring surface of the light guide and the preferably flat measuring surface of the second ATR body have a continuous, uniform measuring path, within which a measuring beam preferably at least twice, preferably at least four times, attenuated total reflectable, i. can interact with an adjacent, in particular the measuring surface only continuously wetted, liquid medium. In general, 2 to 20 total reflections along a measurement path are sufficient to collect enough data for the evaluation.
  • the refractive index of the first and second ATR bodies is greater than or equal to 1.5.
  • the ATR bodies used in the analyzer according to the invention are also referred to in the art as ATR crystals, although these systems are not necessarily in crystal form. Accordingly, e.g. Also, sintered silver chloride is a functional ATR body.
  • the material used for the ATR body used is any material that is transparent to the radiation used, in particular for electromagnetic radiation in the mid-infrared range, and which, moreover, is highly refractive or is high refractive index and has a refractive index greater than that of air and / or as that of a medium to be analyzed or analyzed with the infrared measuring device according to the invention.
  • Suitable materials for the ATR body include diamond, sapphire, cadmium telluride, thallium bromide iodide, silicon, germanium, zinc selenide, zinc sulphide, magnesium difluoride, cesium iodide, silver chloride, calcium difluoride, potassium bromide or sodium chloride. It will be apparent to those skilled in the art that certain of the foregoing materials, unless accompanied by a suitable
  • an ATR body of sodium chloride is made of a transparent material suitable for infrared radiation.
  • pension material in particular a polymer material, with a refractive index preferably> 1.5, in particular made of polyethylene, resorted.
  • this ATR body may otherwise be in any desired geometry, as long as it allows an incident beam to be adjusted in such a way that this has been totally reflected at least twice attenuated before its exit from the ATR body.
  • with six or seven such total reflections already optimal analysis results in terms of sensitivity, accuracy and speed are also achieved for aqueous multicomponent systems.
  • a measuring path is understood as meaning that section of the ATR body which comes into contact with the medium to be analyzed and which is available overall for attenuated total reflections.
  • a sample holder with a receiving device is preferably used, which is designed to receive at least one, in particular exactly one, liquid drop.
  • this Aufhahmevorraum is designed to receive a quantity of liquid having a volume in the range of 0.1 to 400 .mu.l, in particular from 1 to 50 ul.
  • Amounts of liquid in the nl and ⁇ l range can be readily applied to the receiving devices via suitable pipetting devices.
  • the expert also has automatic pipetting robots available, with which a high mass flow rate is readily achievable. Nor is it of any particular difficulty, a precisely defined
  • Quantity or a predetermined number of drops for example 1, 2, 3, 4, 5, 6 or 7 drops to be applied in the receiving device.
  • a holding device is preferably provided in the sample holder, in which the side of the liquid to be measured lying opposite the support surface lies at least in regions above the edge of the receiving device in a substantially vertical orientation of the sample holder.
  • This constellation can be adjusted by the skilled person readily depending on, for example, the capacity and the diameter of the Aufhahmevorplatz and the polarity of liquid and inner wall material of the Aufhahmevorraum.
  • the surface tension of the quantity of liquid present in the receptacle causes liquid also to protrude beyond the edge of the receptacle and thus become accessible even to such a measuring surface of an ATR body which is larger in size than the circumference of the receptacle.
  • the embodiment described above finds particular application in very small sized Aufhahmevoriquesen, for example, with a volume in the range of 1 to 100 ul.
  • the amount of liquid present in the Aufhahmevorraum be sized so that it does not protrude beyond the edge of this Aufhahmevorraum out.
  • the ATR body is brought to the surface of the liquid present in the Aufhahmevorraum so long, or vice versa, the Aufhahmevortechnisch to the ATR body until the measuring surface of the ATR body is wetted by the liquid.
  • the edge of the receiving device is thus dimensioned in height and / or extent or extent that liquid protrudes beyond the edge due to surface tension phenomena and is available for contact with the measuring surface of an ATR body or that the ATR body has a smaller size Expansion has as the edge and thus can be introduced into the Aufhahmevortechnisch.
  • the device according to the invention accordingly allows the use of a smallest possible dimensioned ATR body with which an IR measurement is still possible.
  • the material costs can be reduced considerably and, on the other hand, very small-dimensioned systems can be designed, which can be used without difficulty for, for example, point of c ⁇ re measurements.
  • the device according to the invention only very small amounts of liquid conditions are required, for example, in the blood test, the required amount of blood can be drastically reduced. So far, for usual blood tests about 5 ml of blood required, it now comes out for the same investigation with only about 5 ul.
  • the amount of waste products that regularly accumulates when larger amounts of blood are withdrawn can be significantly reduced. For example, it is not uncommon for conventional clinics to take about 1,200 samples per day.
  • measuring surface at least partially a, in particular hydrophobic, coating which is transparent to the measuring radiation.
  • the coating has a thickness which is less than the wavelength of the measuring radiation used, it is possible to fall back on any coating material transparent to the measuring radiation.
  • the thickness or thickness of the coating is not critical if the coating material is one for an ATR body, that is to say has approximately the same refractive index as that of the particular ATR body used.
  • an ATR material such as zinc sulfide or zinc selenide, which is particularly preferably used for the ATR bodies used according to the invention, can be provided with a layer of diamond. Obtained in this way a provided with a highly resistant and inert coating ATR body.
  • the diamond layer is particularly preferably applied according to a method described by HJ Neubert in Optics, February 2002, page 11.
  • a temperature in the range of 15,000 to 20,000 ° C is generated in the vicinity of a surface by means of a carbon dioxide laser with a power of about 6 to 7 kW.
  • a plasma is generated.
  • gaseous hydrocarbons for example methane
  • free carbon atoms are formed which deposit on a substrate, in this case, for example, on a flat boundary surface of the second ATR body, forming a diamond layer of very low thickness to let.
  • the coating described above especially for materials for ATR body, the toxic, soluble, z. B. in the sample medium, and / or sensitive to mechanical stress.
  • Particularly suitable for the measuring device according to the invention are ATR bodies of zinc sulfide or zinc selenide, which are provided with a coating, in particular a diamond coating.
  • the coating of the ATR body of the device according to the invention has a thickness which is less than the, preferably half, wavelength of the used mfrarot measuring radiation, in particular over a thickness in the range of about 2 nm to about 250 nm Thickness of the coating is particularly preferably in a range of about one quarter of the measuring wavelength. Furthermore, it is recommended to use a coating which is homogeneous in terms of thickness and composition and has the smoothest possible surface. It is advantageous if the unevenness in the coating does not exceed the extent of about one quarter of the measuring wavelength on average. Suitably, recourse is made to those coatings which do not reflect the measurement radiation, even partially.
  • the coating performs a dual function by protecting the test medium from contamination with, possibly toxic, ATR body material as well as the ATR body against mechanical damage on the one hand. Coatings that do not allow diffusion of water molecules through this layer to the ATR body have proven particularly useful in the study of aqueous systems.
  • a layer of a transparent or translucent plastic in particular polyethylene.
  • the ATR body materials which can be provided with a coating are selected from sapphire, cadmium telluride, thallium bromide iodide, silicon, germanium, zinc sulphide, zinc selenide, magnesium difluoride, cesium iodide, Silver chloride, calcium difluoride, potassium bromide and sodium chloride, wherein zinc selenide and
  • Zinc sulfide are preferred.
  • the Aufhahmevoriquesen are in the form of wells and are preferably combined in a field or array of Aufhahmevortechniken.
  • the suction device can also be part of a device for taking blood, containing at least one capillary gap, or be such.
  • blood sampling devices are known for example as so-called test strips for diabetics.
  • sample holder a support, for example in the form of a plate, e.g. similar to a microtiter plate, comprising at least two, preferably at least eight, and in particular a plurality of Aumahmevorrichronne.
  • the rows of Aufhahmevoriquesen succeed each other and form, for example, an array of receiving wells, as it is known for example from microtiter plates.
  • the rows of receptacles may also be arranged in a star-shaped, circular, spiral or linear manner on the sample plate.
  • the corresponding Advicehahmevoriquesen are located approximately equidistant from the center and are accordingly on the same circumference. In this way, by rotation of the sample holder, the ATR body can be positioned quickly, unproblematically and reliably over a new suction device.
  • the sample holder preferably has a planar geometry but may be e.g. also be constructed stepwise.
  • the receiving devices may be mounted on the sample holder or may be integral therewith.
  • the sample holder has openings or inlets, in each of which a receiving device, in particular reversibly, can be inserted or inserted.
  • a receiving device in particular reversibly, can be inserted or inserted.
  • Such Advicehahmevoriquesen be included, which are a device for collecting blood, containing at least one capillary gap, or are part of such a device.
  • the positioning device is designed to move the measuring surface of the ATR body in the direction of an amount of liquid present in a receiving device of the sample holder or away from and / or around the sample holder in the direction of the measuring surface or to move this away, in particular to establish or cancel a contact.
  • the infrared light source comprises one or more quantum cascade lasers.
  • Quantum cascade lasers which are suitable for the device according to the invention are known, for example, from EP 0 676 839 A and from US Pat. No. 5,509,025, in which the basic mode of operation and the structure thereof are described.
  • quantum cascade lasers which emit electromagnetic radiation in the mid-infrared range.
  • such quantum cascade lasers come into consideration, which emit only a defined frequency, in particular from the mid-infrared range, as well as those which emit two, three, four, five or more frequencies. frequencies, in particular from the mid-infrared range, can radiate.
  • these Infrarotmeßvorraumen not only with one, but with two or
  • a quantum cascade laser which is capable of emitting electromagnetic radiation of at least two different frequencies, in particular from the mid-infrared range, or if several quantum cascade lasers are used side by side in such a measuring device, the electromagnetic radiation, in particular if it has different frequencies, emitted at the same time or almost simultaneously or in chronological order.
  • the electromagnetic radiation in particular if it has different frequencies, emitted at the same time or almost simultaneously or in chronological order.
  • it is possible to have multiple ingredients present in a sample in a very short time, i. simultaneously or almost simultaneously. Almost simultaneously or simultaneously within the meaning of the present invention means that signals are radiated so slightly offset in time, that no significant differences from the absorption signals detected at the same time emitted radiation can be seen from the respective detected absorption signals.
  • the quantum cascade laser used emits electromagnetic radiation in the form of pulses having a defined duration and / or intensity.
  • This pulse duration and / or intensity can be freely selected within wide ranges and can be used to generate optimized spectrometric examination conditions for each ingredient to be examined.
  • pulse durations of different lengths and / or intensities of different intensities can be selected depending on the frequency of the emitted electromagnetic radiation. For example, ingredients with weakly absorbing chromophores can be given pulses of longer pulse duration, while strong ones Absorbing substances are very short pulse durations sufficient to detect a satisfactory signal can.
  • pulse patterns and / or intensity patterns can be used which are tailored to the respective analysis problem.
  • the frequencies, pulse durations and / or intensities can be predetermined in such a way that the type and extent of the detected signals can be determined with the aid of an evaluation unit, in particular a computer-aided evaluation unit. can easily determine which ingredients are present in which concentrations, in the examined sample.
  • These patterns of pulse sequence, pulse length, and / or pulse intensity of two or more frequencies may in turn be used to generate particular patterns of response signals characteristic of particular compositions.
  • the multiplexer used according to the invention additionally controls or regulates the intensity of the respective measuring radiation, in particular as a function of the respective measuring problem.
  • optical multiplexers thus coordinate the radiation source and the detector and tune them to each other.
  • multiplexers switch on and off measuring radiation according to a pulse sequence tailored to the measurement problem, in particular with wavelength-dependent modulation of the intensity, and thus control the light source.
  • the detected signals are preferably evaluated using known methods such as factor analysis, multiple least square algorithms or neural network analysis.
  • computer-based evaluation units are used regularly.
  • a characteristic feature associated with the device according to the invention is that in multicomponent mixtures, several ingredients can be determined simultaneously and qualitatively next to each other.
  • infrared light sources which emit a continuous spectrum.
  • Such light sources are known to those skilled in the art, for example as Nernst pins, which consist essentially of zirconium oxide and rare earth additives, and as so-called Globare, consisting essentially of silicon carbide.
  • the light source is an electrically conductive ceramic in question.
  • light sources can be used which emit over the entire spectral infrared range or only over certain ranges of this spectrum.
  • the positioning device is designed to provide a distance between the measurement surface and the edge of the capture device that is less than 1 mm, preferably less than 15 ⁇ m, in particular without the measurement surface touching the edge.
  • the ATR body and fluid sample in the receiving device are only moved toward one another relative to one another until wetting of the measuring surface of the ATR body with the liquid takes place by capillary action.
  • the distance between the measuring surface and the edge can generally be varied within wide limits.
  • the device according to the invention is regularly equipped with an infrared measuring device comprising the ATR body, at least one infrared light source, at least one detector and at least one evaluation unit. Integrated into the evaluation unit or separately therefor may be a display unit. As detectors for the registration of the measuring radiation can be used on all current systems used in Infrarotmeßvorraumen systems.
  • An evaluation unit in the sense of the present invention may also comprise a data memory and / or a display unit, e.g. a pen or a screen. Of course, these elements can also be present separately.
  • An embodiment of the invention provides that the evaluation unit is suitable for evaluating the signals recorded by the detector by means of Fourier transformation.
  • the interferogram recorded in the detector of the FT-IR spectrometry device which records a superposition of all wavelengths occurring in the spectrum, is computer-assisted in the evaluation unit by Fourier transformation into the frequencies of the individual oscillations. Details of the Fourier transform are e.g. at N.B. Colthup, L.H. DaIy, S.E. Wiberley, Introduction to Infrared and Raman Spectroscopy, Academic Press, San Diego, 1990, which is hereby incorporated by reference.
  • the FT-IR measuring device several ingredients can be determined at substantially the same time with high sensitivity, speed and wavenumber precision.
  • the device according to the invention may comprise at least one filling device for filling and / or rinsing at least one, in particular all Aufhahmevor- directions, and / or at least one drying device for drying at least one emptied and / or rinsed Aufhahmevorraum.
  • the positioning device in such a way that a large number of picking devices present on the sample holder or the liquid to be measured therein, in particular automatically, approach the measuring surface of the ATR body and, after the measurement has been taken, move away again to lead.
  • the inventive device is suitable for, in particular simultaneous, qualitative and / or quantitative determination of IR-active ingredients in aqueous or non-aqueous liquids.
  • aqueous liquids to be measured e.g. in question body fluids such as blood, serum, blood plasma, saliva, sweat, semen, urine, lymph, spinal fluid and interstitial body fluid and beverages such as beer, wine, milk, milk products, fruit juice, spirits or soft drinks.
  • question body fluids such as blood, serum, blood plasma, saliva, sweat, semen, urine, lymph, spinal fluid and interstitial body fluid and beverages
  • beverages such as beer, wine, milk, milk products, fruit juice, spirits or soft drinks.
  • the device according to the invention can be used e.g. medically relevant constituents such as glucose, albumin, total protein, urea, creatinine, hemoglobin, hematocrit, uric acid, phosphate, acetoacetate, acetone, hydroxybutyric acid, sulfur, in particular in the form of sulfate, cholesterol and / or tricglycerides and their concentrations simultaneously side by side be determined with high accuracy and high throughput.
  • medically relevant constituents such as glucose, albumin, total protein, urea, creatinine, hemoglobin, hematocrit, uric acid, phosphate, acetoacetate, acetone, hydroxybutyric acid, sulfur, in particular in the form of sulfate, cholesterol and / or tricglycerides and their concentrations simultaneously side by side be determined with high accuracy and high throughput.
  • creatinine in urine hemoglobin in blood, hematocrit in blood, uric acid, phosphate, acetoacetate, acetone, hydroxybutyric acid, sulfur, especially in the form of sulfate, can be detected quantitatively in urine.
  • non-aqueous liquids for example systems comprising at least one IR-active substance dissolved in an organic solvent
  • the device according to the invention can be used, for example, in the investigation of dye or lacquer patterns.
  • the object underlying the invention is further achieved by a method for the quantitative and / or qualitative determination of IR-active ingredients in aqueous or non-aqueous liquids using a device according to the invention, comprising the steps a) introducing a liquid sample, in particular at least one drop, of the liquid sample to be measured into a first receiving device of the sample holder; b) contacting the measuring surface of the first or second ATR body with the surface of the liquid sample to be measured in a first receiving device so that covering an area as large as or greater than the optically active ATR area; c) measuring the liquid sample containing at least one IR-active ingredient using an infrared measuring device, comprising passing infrared radiation emanating from an infrared light source through the ATR body and detecting the signals emanating from the ATR body in at least one detecting unit, and d) removing the measuring surface of the ATR body from the surface of the liquid, after step c) and / or d) the detected signals in an evaluation unit for the purpose of
  • a plurality of IR-active ingredients present in the liquid sample are preferably measured simultaneously and determined substantially qualitatively and / or quantitatively at the same time.
  • five, ten or even 20 ingredients can be readily determined qualitatively and / or quantitatively substantially simultaneously.
  • steps a) to d) can be switched or repeated as often as desired, with each new step a) preferably giving up a certain amount of liquid of a new sample to be examined.
  • a further development is characterized by the following additional steps: e) cleaning the measuring surface of the ATR body, f) contacting the cleaned measuring surface of the ATR body with the surface of a liquid present in a second recording device, g) measuring the liquid sample, containing at least one IR-active ingredient using an infrared measuring device, comprising passing IR radiation emanating from an infrared light source through the ATR body and detecting the signals emanating from the ATR body in at least one of them
  • Detection unit and h) removing the measuring surface of the ATR body from the surface of the liquid, wherein after step g) and / or h) the detected signals in an evaluation unit for the purpose of qualitative and / or quantitative determination of at least one ingredient of the liquid sample are evaluated ,
  • the steps e) to h) are repeated at least twice and in the case of high sample throughput series, wherein a new liquid sample to be measured is present in the receiving device in each case.
  • the cleaning step e) can take place, for example, in such a way that in a first step e ⁇ a Aufhahmevortechnisch on a sample holder with a rinsing or cleaning liquid is filled, which then in a second step e 2 ) with the measuring surface of the ATR body reversibly brought into contact.
  • additional method steps are provided which can be followed, for example, by steps d), e) and / or h): i) introducing a reference liquid, in particular at least one drop of the reference liquid, into a third receiving device of the sample holder, j) contacting the measuring surface of the first or second ATR body with the surface of the reference liquid to be measured in the third sampling device, so that preferably one K) measuring the reference liquid containing at least one IR-active reference ingredient using an infrared measuring device, comprising passing IR radiation emanating from an infrared light source the ATR body and detecting the signals emanating from the ATR body in at least one detection unit, and
  • step k) 1) removing the measuring surface of the ATR body from the surface of the reference liquid, wherein after step k) and / or 1) the detected signals in an evaluation unit for the purpose of qualitative and / or quantitative determination of at least one reference ingredient of the liquid sample are evaluated.
  • the detection process in the sense of the present invention generally also includes the storage of the recorded signals or the recorded raw data set in a suitable data memory, so that these data are henceforth available for further evaluation.
  • Suitable detection units and devices for storing larger amounts of data are well known to those skilled in the art.
  • the measuring surface of the ATR body can be successively brought into contact with the surface of the liquid samples to be measured.
  • the measuring surface of the ATR body is first cleaned after each contact with a liquid to be measured, preferably by bringing it into contact with the cleaning or rinsing liquid present in a receiving device.
  • the measuring surface of the ATR body is brought into contact with a reference liquid at least once before recording the measurement of liquid samples present in the recording devices and / or during the measurement of a multiplicity of such liquid samples. In this way, a calibration or recalibration can be made.
  • the reference liquid also serves as a rinsing liquid, whereby both the material as well as the process cost can be further reduced.
  • the Aufhahmevoriquesen present on a sample holder are first filled with the liquid samples to be measured and optionally the rinsing and / or reference liquid, i. the steps a) and optionally e ⁇ ) and / or i) are preceded. Subsequently, the sequences of steps b) -c) -d) and optionally f) -g) -h) and / or j) -k) -l) are then used. These sequences can be repeated as desired or combined with each other.
  • recording devices can be filled alternately according to the following pattern: measuring liquid, rinsing liquid, ..., measuring liquid, rinsing liquid, etc.
  • the sequence can provide: reference liquid, rinsing liquid, measuring liquid, rinsing liquid,..., Reference liquid, measuring liquid, Rinsing fluid, etc.
  • the respective liquids can be combined in any desired sequence.
  • the measurement result can be further optimized by the fact that the measuring surface of the ATR body after contacting with the present in a Trohahmevortechnik to be measured liquid with a cleaning liquid, which is another, in particular adjacent, Aufhahmevortechnisch on the sample holder is brought into contact.
  • the measuring liquid is generally measured only for a period of 1 to 60 seconds. Measuring times of 2 to 30 seconds are usually sufficient for a sample, even measuring times in the range of 1 to 3 seconds are readily available. Due to the small sample volumes and the very short measuring time per sample, sample holders can easily be used which have more than 100, for example 500 to 1000 or 1000 to 2000, recording devices.
  • Particularly reliable measurement results occur when a multiplicity of liquid drops present on a sample holder in holding devices are measured successively, in particular automatically. It is also of particular advantage that at least one liquid to be measured represents a reference sample. Moreover, it can be provided that the measuring surface of the ATR body is subjected to a drying step after a measuring step and / or a cleaning step.
  • the present invention was based on the surprising finding that a very high mass throughput can be achieved even with a non-contact optical measuring method for the determination of ingredients in liquids, even when using sample volumes in ⁇ l range, without the accuracy suffers ,
  • a further advantage is that a sample preparation or evaporation or drying of the sample is eliminated and it is also not dependent on the use of analysis reagents.
  • the method according to the invention allows a linear calibration over a very wide measuring range. Also eliminates the generally necessary dilution series.
  • the miniaturized design of the ATR unit and the concomitant low measurement volumes allow the use of the method according to the invention in neonatal and paediatrics, where so far mainly test strip methods are used due to the small amount of blood available.
  • these methods are much too inaccurate and also not very reliable.
  • relatively large amounts are required, for example, in the measurement of blood by conventional methods, amounts in the lower ⁇ l range are already sufficient for reliable measurements by the method according to the invention.
  • Another advantage here is that only very small volumes of reference solution is required, which compared to conventional methods brings a significant cost savings. In any case, it can be ensured with the method according to the invention that the number of reference measurements is no longer reduced beyond the required level to the detriment of accuracy.
  • a further advantage is that with the device according to the invention, the liquid samples to be measured can be measured very promptly for sampling. This is significantly noticeable, for example, in the determination of the glucose content in the blood. If, for example, a measurement is delayed, a too low glucose value is determined due to the glucose breakdown continuing in the blood sample. As a rule, it is sufficient to prepare the blood samples to be measured, to provide them with an anticoagulant. Optionally, agents that inhibit glucose degradation, such as sodium fluoride, may also be added. Further embodiments of the invention will be described in detail with reference to the following figures, without the invention being restricted to these particular embodiments. Show it
  • Figure 1 is a cross-sectional view of a sample holder and an ATR body of the device according to the invention
  • Figure 2 is a cross-sectional view of a sample holder and an ATR body of an alternative embodiment of the device according to the invention
  • Figure 3 is a plan view of a sample holder of an embodiment of the device according to the invention.
  • Figure 4 is a plan view of an alternative sample holder for an inventive
  • Figure 5 is a plan view of an alternative sample holder for an inventive
  • FIG. 1 shows a sample holder 2 of a device 1 according to the invention and an ATR body 4 in cross-section.
  • the sample holder 2 has essentially a flat base plate 6 mounted on the trough-shaped Aufhahmevorraumen 8 or integrated.
  • the Aufhahmevortechniken 8 stand out with its edge 10 from the plane spanned by the base plate level and preferably form the highest vertical point in the Aufhahrnevorraum.
  • In the trough-shaped Aufhahmevorraum 8 is to be measured liquid drop 12.
  • An ATR body 4 is positioned in such a way above the Aufhahmevortechnik 8 that its measuring surface 14 can come into contact with the liquid droplet 12.
  • the drop of liquid 12 rests on the support surface 16 of the Aufhahmevorraum 8 and is limited by the peripheral edge 10. Due to surface tension phenomena, the liquid drop 12 protrudes in vertical alignment beyond the edge 10 and is thus available for contact with the measuring surface 14 of the ATR body 4.
  • the sample holder 2 is preferably moved in the direction of the arrow to a fixed ATR body 4 by means of a positioning device 18 and then moved away again.
  • the positioning device 18 is capable both of aligning the sample holder in the vertical direction, as well as after completion of the measurement of the Drop 12, ie after the sample holder 2 has been lowered, continue to move it horizontally until the next, preferably adjacent, suction device containing a drop of liquid 20 comes to lie directly below the measuring surface 14 of the ATR body 4. Then, the positioning device 18 can approach the sample holder again to the measuring surface, but without it comes to a contact of the measuring surface 14 with the edge 10 of the Aurhahmevorraum 8.
  • the embodiment of the inventive analysis device 1 according to FIG. 2 differs from that according to FIG. 1 solely in that not an ATR body 4 but a multiplicity of ATR bodies 4, 4 ", 4" 'are present. In this way, the mass flow rate can be increased considerably again.
  • FIG. 3 shows a plan view of a sample holder 2 of a device 1 according to the invention.
  • This sample holder 2 has a rectangular base area with a total of 96 Aufhahmevortechniken 8, which are present in an 8 x 12 field (indicated).
  • the field which is moved up first to the measuring surface of the ATR body may contain a drop of a reference liquid KA, with which the device is first calibrated.
  • the following in the series Aumahmevorraum then contains a cleaning liquid R 5 to which in the following Aufhahmevoroplasty a first liquid to be measured M follows.
  • Figure 4 shows an alternative sample holder 2, on which the Aufhahmevortechniken 8 are arranged on a circumference.
  • the Mean Time to Browse Ratio For the movement of Aufhahmevortechniken in the desired position is sufficient for an angle motor.
  • it is of particular advantage that it can easily be used fully or semi-automatically for a continuous operation mode.
  • already measured measuring liquid or once used cleaning or reference liquid from their receiving devices disposed of and optionally cleaned and then again with liquid to be measured or cleaning or Reference liquid are filled.
  • FIG. 5 shows an alternative embodiment of a sample holder 2.
  • rows of receiving devices 8 are arranged substantially in a star shape.
  • a positioning can be effected by a rotation of the sample holder under an ATR body.
  • sample holders in any geometry and arrangement of the individual receiving devices for the device according to the invention can be used.

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Abstract

La présente invention concerne un dispositif pour réaliser la détermination qualitative et/ou quantitative de substances à activité IR contenues dans des liquides aqueux ou non aqueux, le dispositif comprenant: a) au moins un premier corps ATR qui se présente sous la forme d'une guide d'onde optique comprenant une surface de mesure qui vient en contact avec le liquide à mesurer, qui est transparent ou semi-transparent au rayonnement de mesure, et qui présente un indice de réfraction qui est supérieur à celui du liquide adjacent à la surface de mesure lors de la mesure, au moins un second corps ATR qui comprend au moins une surface limite plane comprenant une surface de mesure qui vient en contact avec le liquide à mesurer, qui est transparent ou semi-transparent au rayonnement de mesure, et qui a un indice de réfraction qui est supérieur à celui du liquide adjacent à la surface de mesure lors de la mesure; b) au moins un porte-échantillon qui contient au moins un dispositif de réception destiné au liquide à mesurer, qui présente une surface d'appui destinée au liquide, et un bord, et qui a des dimensions qui permettent de garantir un contact entre la surface de mesure du premier ou du second corps ATR avec une surface du liquide à mesurer, présent dans le dispositif de réception; et c) au moins un dispositif de positionnement qui sert à mettre en contact réversible la surface du liquide et la surface de mesure du premier ou du second corps ATR. L'invention a également pour objet un procédé pour réaliser une détermination quantitative et/ou qualitative de substances à activité IR présentes dans des liquides aqueux ou non aqueux, en se servant du dispositif de l'invention, le procédé comprenant les phases suivantes: a) introduction d'un échantillon de liquide, en particulier d'au moins une goutte de l'échantillon de liquide à mesurer, dans un premier dispositif de réception du porte-échantillon; b) mise en contact de la surface de mesure du premier ou du second corps ATR avec la surface du liquide à mesurer présent dans le premier dispositif de réception de sorte qu'une surface de taille supérieure ou égale à celle de la surface ATR optiquement active, se trouve recouverte; c) mesure de l'échantillon de liquide contenant au moins une substance à activité IR, par utilisation d'un dispositif de mesure infrarouge, comprenant le passage de rayonnement IR sortant d'une source de lumière infrarouge, par le corps ATR, et la détection des signaux émis par le corps ATR dans au moins une unité de détection; et d) retrait de la surface de mesure du corps ATR, de la surface du liquide, les phases c) et/ou d) étant suivies de l'évaluation des signaux détectés dans une unité d'évaluation afin de réaliser la détection qualitative et/ou quantitative d'au moins une substance contenue dans l'échantillon de liquide.
PCT/DE2006/001771 2005-10-10 2006-10-10 Dispositif et procede pour realiser la determination qualitative et/ou quantitative de substances a activite ir contenues dans des liquides WO2007041999A1 (fr)

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DE102005048807A DE102005048807B3 (de) 2005-10-10 2005-10-10 Vorrichtung für die qualitative und/oder quantitative Bestimmung von IR-aktiven Inhaltsstoffen in Flüssigkeiten sowie ein Verfahren zur qualitativen und/oder quantitativen Bestimmung von IR-aktiven Inhaltsstoffen in Flüssigkeiten
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DE102005048807B3 (de) * 2005-10-10 2006-11-16 Johann Wolfgang Goethe-Universität Vorrichtung für die qualitative und/oder quantitative Bestimmung von IR-aktiven Inhaltsstoffen in Flüssigkeiten sowie ein Verfahren zur qualitativen und/oder quantitativen Bestimmung von IR-aktiven Inhaltsstoffen in Flüssigkeiten
GB0706398D0 (en) * 2007-04-02 2007-05-09 Univ Hospital Of North Staford Improvements in and relating to copd determination
AT512291B1 (de) * 2012-02-20 2013-07-15 Anton Paar Gmbh Verfahren und vorrichtung zur bestimmung des co2-gehalts in einer flüssigkeit
DE102014108424B3 (de) 2014-06-16 2015-06-11 Johann Wolfgang Goethe-Universität Nicht-invasive Stoffanalyse
US10876965B2 (en) 2015-12-09 2020-12-29 Diamontech Ag Apparatus and method for analyzing a material
CN108369183B (zh) * 2015-12-09 2022-08-19 迪亚蒙泰克股份有限公司 用于分析材料的装置和方法

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EP0516481A2 (fr) * 1991-05-31 1992-12-02 Donald W. Sting Elément pour la spectroscopie à réflexion interne avec une surface très petite pour contacter l'échantillon
DE4333560A1 (de) * 1993-10-01 1995-04-06 Bayer Ag Vorrichtung zur kontinuierlichen spektroskopischen Analyse nach dem Prinzip der abgeschwächten Totalreflexion
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