EP2861973A1 - Procédé, dispositif et appareil de mesure portatif pour la détection de produits de dégradation de molécules biologiques dans des couches d'un système de couches - Google Patents

Procédé, dispositif et appareil de mesure portatif pour la détection de produits de dégradation de molécules biologiques dans des couches d'un système de couches

Info

Publication number
EP2861973A1
EP2861973A1 EP13740203.8A EP13740203A EP2861973A1 EP 2861973 A1 EP2861973 A1 EP 2861973A1 EP 13740203 A EP13740203 A EP 13740203A EP 2861973 A1 EP2861973 A1 EP 2861973A1
Authority
EP
European Patent Office
Prior art keywords
radiation
layer
degradation products
excitation
layer system
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP13740203.8A
Other languages
German (de)
English (en)
Inventor
Holger Lausch
Michael Arnold
Michael Brand
Joseph Maria Regina Delhaes
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.)
Fraunhofer Gesellschaft zur Foerderung der Angewandten Forschung eV
Original Assignee
Fraunhofer Gesellschaft zur Foerderung der Angewandten Forschung eV
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 Fraunhofer Gesellschaft zur Foerderung der Angewandten Forschung eV filed Critical Fraunhofer Gesellschaft zur Foerderung der Angewandten Forschung eV
Publication of EP2861973A1 publication Critical patent/EP2861973A1/fr
Withdrawn legal-status Critical Current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
    • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G01N33/521—Single-layer analytical elements
    • 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
    • G01N21/88—Investigating the presence of flaws or contamination
    • G01N21/94—Investigating contamination, e.g. dust
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/575—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57525—Immunoassay; Biospecific binding assay; Materials therefor for cancer of the liver or pancreas
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2201/00—Features of devices classified in G01N21/00
    • G01N2201/06—Illumination; Optics
    • G01N2201/061—Sources
    • G01N2201/06113—Coherent sources; lasers

Definitions

  • the invention relates to a method for detecting degradation products of biological molecules of certain properties by excitation of the biological molecules by means of radiation and the detection and evaluation of a remitted due to the excitation radiation of the molecules, as is known generically from the document DE 103 15 541 A1.
  • the invention also relates to devices for detecting biological molecules and devices for carrying out the method.
  • biodegradation products of biological molecules are intermediates or end-products that have been synthesized by organisms.
  • Degradation products are the result of metabolic processes or caused by organisms degradation of chemical compounds, especially biological molecules.
  • a disadvantage of the mentioned method is that the excitation of the biological molecules takes place on the surface of the food and for this purpose the food must be present unpacked during an examination.
  • the invention is therefore based on the object to propose a possibility for the detection of degradation products of biological molecules in layers of a layer sequence. It is also an object of the invention to provide a device for detecting degradation products of the biological molecules. Furthermore, it is an object to propose a transportable device for carrying out the method.
  • the object is achieved by a method for detecting degradation products of biological molecules in layers of a layer system in which no chemicals are introduced into the layer system to enable the detection of the degradation products and in which the degradation products of biological molecules are exposed to an excitation radiation and one by the excitation radiation detects conditioned radiation of the degradation products and determines a concentration of the degradation products based on a comparison of the measured values of the radiation with reference data, achieved by providing a layer system which comprises a transmission layer transparent to the excitation radiation and the radiation and at least one further degradation product containing Receiver layer comprises, wherein the excitation radiation is directed through the passage layer into the receiver layer.
  • Degradation products are molecules that can arise when biodegradation occurs.
  • the invention relates to such molecules that show autofluorescence.
  • Degradation products can also be biological molecules (biological active molecules) or biogenic compounds (molecules).
  • the terms "degradation product (s)" and “degradation products of biological molecules” are used synonymously.
  • a radiation caused by the excitation radiation (hereinafter also referred to as "radiation") of the degradation products is detected, whereby it is not excluded that a, mostly low, fraction of detected radiation results not from degradation products but from other sources such as impurities
  • the measured values of the radiation are compared with reference data Comparison of the measured values with the reference data determines an ("equivalent") concentration of the degradation products, which does not reflect the actual concentration, but just like an actual concentration gives an indication of the current state of the coating system
  • An indication that the state of the layer system is outside a permissible range, for example, is corrupted can advantageously be carried out without introducing chemical substances into the layer system by means of which a detection of the degradation products is supported or This makes detection possible without changing the structure of the layer system, the chemical composition of the layer system or the specific (micro) climatic conditions of the layer system It is very advantageous that the layer system does not have to have a precisely defined structure and no predetermined dimensioning.
  • the pieces of meat can have different thicknesses.
  • a fleece insert may optionally be present in addition to a film wrapping.
  • the individual layer systems can be examined by means of one and the same inventive method.
  • chemical substances are, in particular, molecules or elements which are suitable as markers, the addition of which enables the detection of molecules to be detected.
  • the method according to the invention can be carried out without the layer system having to be specifically assembled for detection, as is necessary, for example, when examining histological sections. Furthermore, the method according to the invention can be carried out without the layer system having to be changed, in particular completely or partially dismantled.
  • the example of the detection of degradation products on packaged foodstuffs means that a packaging film does not have to be removed to carry out the process.
  • the method is also advantageously suitable for successive detections of degradation products of biological molecules of one and the same layer system that are spaced apart from each other in time (for example several hours or days).
  • the layer system is vertically aligned. A first layer is therefore at the top, while the further layers of the layer system are arranged successively below the first layer. If the layer system is oriented differently in space, the description is to be understood accordingly.
  • the passage layer is preferably the uppermost layer of the layer system. It may, for example, be a plastic packaging film or a plastic lid, as commonly used in the packaging of foodstuffs such as meat and sausage products, fish, fruit or vegetables.
  • the term "transparent” should also include the term "translucent”.
  • a receiver layer is a layer system layer in which the presence of degradation products of biological molecules is to be investigated and a concentration of any biological molecules present should be determined.
  • a layer to be examined is therefore a receiver layer even if it contains no degradation products.
  • Crucial is only the procedural investigation of the relevant layer.
  • the acquired radiation is evaluated as a whole, without highlighting its origin.
  • a receiver layer containing the degradation products may be a first film layer which may be present, for example, as a thin layer (film) of a condensate immediately on the underside of the via layer.
  • a gas layer a second film layer on the surface of the food, the food itself and a bottom layer, for.
  • the bottom layer can again be surrounded with a film, for example with edge regions of a film forming the passage layer, downwards.
  • the method is applied to a layer system comprising a through layer and a first film layer immediately adjacent thereto.
  • the first film layer is preferably aqueous and may contain organic compounds, e.g. As biogenic amines.
  • concentration refers both to a determination of a specific concentration specification within the scope of fault tolerances or merely to a comparison with a previously determined limit value. In the second case, it is only checked whether the limit value has been exceeded with sufficient probability. Such an embodiment of the method according to the invention is sufficient for a binary decision such as the purchase or non-purchase of the food.
  • the determination of a specific concentration indication and the comparison with a limit value can be combined.
  • the excitation radiation is directed onto the receiver layer so that the radiation caused by the excitation radiation in the receiver layer can be detected.
  • a beam path along which the excitation radiation is directed into the receiver layer to be examined and a beam path along which the radiation is directed towards a means for detecting the radiation e.g. As a camera, a spectroscope or a photomultiplier, propagates, so be directed into the layer system that intersect the two beam paths in the receiver layer.
  • the excitation radiation is a light of known wavelength.
  • the measured values obtained by the radiation are evaluated colorimetrically (colorimetrically).
  • a second embodiment of the method according to the invention is designed such that the excitation radiation is suitable for exciting fluoresceable biological molecules and their fluoresceable degradation products, the excited molecules emitting a fluorescence radiation.
  • the fluorescence radiation of the excited molecules is the radiation to be detected.
  • the excitation radiation and the radiation emitted by the excited molecules also include radiation spectra.
  • the fluorescent degradation products of the biological molecules are amines.
  • these may be, for example, primary, secondary or tertiary amines.
  • the amines can certain Ptomaine, such.
  • cadaverine (1, 5-diaminopentane) or putrescine, which are caused by the biochemical degradation of proteins and amino acids.
  • amines and their concentration are useful as indicators of the biochemical state of a food.
  • the excitation radiation preferably has at least one wavelength from a wavelength range of 250 to 400 nm.
  • the fluorescence radiation in only one wavelength (measuring wavelength) provided for detection.
  • this measurement wavelength fluorescence radiation is detected only in a narrow wavelength range of, for example, ⁇ 5 nm. This will make the capture of fluorescence radiation that does not originate from molecules to be detected or from other fluorescence radiation sources.
  • Amines have a -C-NH 2 group as a functional group. Excitation of characteristic electronic quantum transitions in the chemical bond region of this functional group causes fluorescence (autofluorescence) at 485 nm. It is therefore advantageous for the reliability of the method according to the invention if the fluorescence radiation in a narrow wavelength range around 485 nm, z. In a range of 475 to 485 nm.
  • the radiation caused by the excitation radiation is excited in the first film layer.
  • enzymatic decarboxylation which may occur during, for example, degradation, i.a. of proteins and amino acids by microorganisms takes place, in addition to biogenic amines such.
  • Ptomainen carbon dioxide and water.
  • Organic substances which are degraded, which are also understood here as foods therefore always have an aqueous layer (eg a second film layer) on their surface.
  • a gas layer will be created over the surface.
  • the aqueous layer and the gas layer are layers of the layer system.
  • the aqueous layer may be a second (on the top of the organic substance) and a third (on the bottom of the organic substance) film layer, which may contain organic compounds.
  • a formation of biogenic amines is prevented or delayed not only by an oxygen-free or oxygen-poor packaging, for example, a food, but also by its cooling. Regardless of whether the cold chain has been ensured continuously, the dew point of the moisture within the more or less dense package due to the cooling is set back so far that the moisture usually on the surface of the food (second film layer) and at the bottom of the passage layer (first film layer) condensed and possibly also crystallized.
  • Both first and second film layers may include organic compounds such as biogenic amines. Due to the Gas layer as transitory transition layer is the distribution of biogenic amines possibly present in the first film layer as an aqueous condensate layer mostly homogeneous, as is the case in the second film layer. This also applies if no gas layer is present.
  • the method according to the invention may be characterized in that the concentration of the degradation products is determined in a layer system in which at least the second film layer, which contains aqueous organic compounds, is present, and by means of the excitation radiation a radiation of the degradation products of the second film layer is excited and the radiation of the degradation products of the second film layer is detected.
  • This radiation can be fluorescence radiation. It can also be a colorimetrically evaluable radiation, wherein such colorimetrically evaluable radiation may also contain fluorescence spectroscopically evaluable wavelengths.
  • the method can also be used to detect biological molecules and / or their degradation products in other layers of a layer system. If these layers are located directly underneath an outer layer of the layer system, a measurement in the near range (near range measurement) will be referred to below, while measurements on layers further away from an outer layer are to be referred to as far range measurements (measurement in the far range).
  • the excitation of the second film layer and the detection of the radiation of the biological molecules of the second film layer are shifted in time to excite the first film layer and to detect the radiation of the degradation products of the first film layer.
  • a determined concentration can be compared with a permissible limit value. If an excess of the limit value is detected with sufficient probability, a warning signal can be generated.
  • the indication signal may be, for example, an optical or acoustic signal or implemented as a combination of different signal types.
  • an impedance of at least one of the layers of the layer system can be measured by impedance spectroscopy and the phase angle of the impedance can be determined. The amounts of the measured impedances can also be determined.
  • impedances of different layers of the layer system can be measured and to determine a shift, i. a difference between the phase angles of the measured impedance.
  • a shift i. a difference between the phase angles of the measured impedance.
  • an impedance can be measured at the uppermost layer and at the lowest layer of the layer system and the phase angle shift as well as absolute value differences can be determined.
  • the impedance spectroscopically examined layers need not be transparent.
  • Electrodes necessary for carrying out the impedance spectroscopy can be brought into contact at the through-layer, at the lowest layer of the layer system or at both said layers in each case from the outside. In both cases, an impedance near the passage layer or the lowest layer is measured (short-range measurement). There can be several electrodes in each case be arranged at the same or different distances, which may also be selectively interconnected to allow a high flexibility of the measurements.
  • the concentration of the degradation products determined on the basis of the radiation and the phase angle shifts and differences in magnitude determined on the basis of the impedance spectroscopic measurements can be examined for congruence with one another. For example, a congruence exists if the condition of a food being studied is found to be sufficiently similar in the different layers. If, on the other hand, the findings of the different strata deviate more than permissible from each other, no congruence is established. If, for example, the impedance spectroscopic measurement at the lowest layer of the layer system points to a fresh food, while the fluorescence spectroscopic measurement of a first film layer indicates that food is already several days old, repacking or replacement of the nonwoven inlay (eg in the case of packaged meat ) be done.
  • the alert signal can then be generated if no congruence is detected.
  • an acknowledgment signal could also be generated if a congruence is detected.
  • a development of the method according to the invention provides that at least one data connection to a virtual memory and arithmetic unit is produced and steps of the method are carried out by means of the virtual memory and arithmetic unit.
  • the virtual memory and arithmetic unit is preferably a dynamically modifiable unit that can be adapted to a current storage and computational requirement, such as a so-called cloud (cloud) in a network such as the Internet.
  • cloud cloud
  • the method according to the invention can also be supplemented to the effect that process-related information is transmitted to a receiving device, for example a database, of a recipient.
  • process-related information can be, for example, information about the location and time of a measurement, information about the measured layer system and about the measured values and results of the evaluations obtained during the execution of the method.
  • procedural information may also include information about a user, such as the person performing the measurement.
  • An identification of a measuring device used to carry out the method can also be transmitted.
  • a recipient may be a third party, such as a supervisor responsible for monitoring compliance with food hygiene legislation.
  • a service provider for. As an Internet portal in the field of consumer protection, or the distributor or provider of the food studied be the recipient. Such an embodiment of the method makes it possible to exchange information with the receiver almost in real time.
  • z. B. allows a health check of packaged food at the point of sale.
  • a time-repeatable condition check of transparently packaged foods is possible.
  • the method can be integrated by appropriately designed devices in storage rooms or in refrigerators, whereby a temporary or permanent monitoring of the condition of the food is possible.
  • the test can on the one hand a qualitative detection of degradation products / biological molecules such as biogenic amines and on the other hand quantitatively determining an intensity of the change in state, eg. B. by the determined concentration include. While the top of a food packaging film is typically transparent, the bottom of the packaging may not be transparent.
  • the object is further achieved by a device for detecting degradation products of biological molecules in layers of a layer system.
  • the device has an excitation source for providing and Radiation of an excitation radiation and a detection unit for detecting the radiation that is caused by the excited Degradations occur (conditional).
  • beam paths of the excitation source and of the detection unit are directed into a common measuring space.
  • a computing unit for controlling the excitation source and for controlling the detection unit and for evaluating the detected radiation is present.
  • the device according to the invention is characterized in that the excitation source is arranged above a layer system located in the measuring space, the layer system having at least one through layer transparent to the excitation radiation and the radiation and subsequently to at least one further layer.
  • the beam path of the excitation source is directed onto the passage layer at a radiation angle in order to excite degradation products in at least one layer located below the passage layer.
  • the detection unit is arranged such that a radiation of the degradation products caused by the excitation radiation can be detected by the detection unit.
  • the detection unit is arranged vertically above the layer system, so that the beam path of the detection unit is directed perpendicular to the layer system. It is advantageous that only the radiation angle must be changed in order to detect radiation from different layers can. Advantageously, unwanted effects can be minimized or hidden by such an arrangement. Thus, excitations and consequent radiation in other than the layer to be examined in a simple manner to neglect, because the beam path of the detection unit is not directed in the region of the other layer or layers in which occur as a side effect suggestions.
  • the beam paths of the excitation unit and of the detection unit preferably intersect in that layer in which the excitation of the degradation products takes place.
  • the emission angles can also be the same, but then the excitation sources can be arranged differently far from the detection unit, respectively of the beam path, so that intersect the beam paths in the layer to be examined.
  • an excitation source which is formed from a plurality of controllable excitation units, each with its own beam paths.
  • the excitation units can each emit an excitation radiation with an individual wavelength and have different opening angles from one another.
  • electrodes for measuring impedances may be present. These can be arranged in pairs or as individual electrodes. The electrodes can be individually controlled and optionally interconnected.
  • the object is further achieved by a portable measuring device for mobile detection by means of the method according to the invention.
  • the measuring device has a receiving tray for receiving a telecommunications device.
  • a receiving tray in the sense of the description may be designed, for example, similar to a holder for mobile phones for use in motor vehicles (hands-free systems).
  • a telecommunications device may be, for example, a mobile phone, a computer, a suitable consumer electronics device or a measuring device. Future technical developments in the field of telecommunications equipment are hereby included in the description.
  • an adapter is preferably arranged, through which a power supply of the measuring device is made by the power source of the telecommunication device. Furthermore, a controllability of the measuring device is made possible by the fact that on the telecommunication device, a program is installed, which is operated by means of the telecommunication device on the adapter. This can be realized by appropriately arranged and designed contacts of the adapter. About the telecommunication device, the data connection of the portable measuring device to the virtual storage and processing unit can be produced.
  • the invention is particularly suitable for a simple, contact and non-destructive testing of food.
  • the inventive method is also useful when one of the film layers is frozen.
  • a typical packaged food is in practice not free of manipulative intervention options by which a formation of degradation products such as biogenic amines and / or an interruption of the statutory cold chain should be covered chemically, visually or by re-labeling and repackaging. While the food control authorities are allowed to unpack the food at random at any time for testing purposes, this is usually not possible for the customer at the point of sale.
  • the invention proposes a method and possibilities of using the method, which enable a customer to make purchase decisions on the basis of objective measurement data. Also, the customer can track the condition of purchased goods over a period of time to make a decision, e.g. B. on the date of consumption of the food or the goods to meet.
  • this is achieved via a mobilizable handheld device for non-destructive and non-contact qualitative detection and quantitative determination of the intensity of biogenic substance groups, by means of which a dual sensor-actuator measurement method is applicable.
  • the quantitative and qualitative determination at the point of sale can not be made alternately over time.
  • the customer has only one test option and the test must be carried out in different layer systems and with unknown through layers.
  • the invention enables a test (detection and concentration determination) independently of the properties of the passage layer.
  • a lid of the can representing the passageway layer may be known by reference measurements and taken into account in subsequent testing of the foodstuff.
  • the phase angle shift in the impedance measurement shows complementary effects in comparison with the change in the fluorescence intensity, such as the zero crossing and a re-increase in the phase angle shift in the emergence and increase of the bioamines (see lecture Sch-40gele http://download.messemuenchen.de / analytica /Presentation/BM_Presentation_020408/Schwaegele.pdf).
  • the inventive method, the device and the portable measuring device are also for a detection of biological molecules in other layer systems usable.
  • a detection and concentration determination of glucose or other hydrocarbons in the epidermis of humans and animals is possible.
  • FIG. 2 shows a highly layered first layer system and an allocation of different procedures for the detection of degradation products of biological molecules to the layers of the layer system
  • FIG. 3 shows a highly layered second layer system and an assignment of different procedures for the detection of degradation products of biological molecules to the layers of the layer system
  • FIG. 4 shows a plan view from below of a first exemplary embodiment of the device according to the invention with a first arrangement of excitation sources, electrodes and a detection unit;
  • FIG 5 is a side view of the first embodiment of a layer system.
  • FIG. 6 shows a plan view from below of a second exemplary embodiment of the device according to the invention of a second arrangement of excitation sources, electrodes and a detection unit;
  • FIG. 7 is a side view of the second embodiment over a layer system
  • FIG. 9 shows a first embodiment of a portable measuring device and its
  • FIG. 10 shows a second embodiment of a portable measuring device and its use
  • 1 is a schematic overview of communication paths between a portable measuring device according to the invention, a virtual memory and arithmetic unit and a receiving device as well as functions and components of the aforementioned components;
  • FIG. 1 shows a decrease in the amount of a phase angle ⁇ determined by means of impedance spectroscopy over the first eight days of storage of pork. This is due, among other things, to a decrease in NADH on the meat. Between the eighth and the tenth day the phase angle ⁇ is approximately constant, which can be explained by an equilibrium situation of the degradation of NADH of the meat and the increase of NADH by a microbial colonization of the meat. From about the eleventh day, the phase angle ⁇ increases significantly again due to an increase in the concentration of microorganisms and their metabolic activities.
  • Fig. 2 is a first layer system 1 with five layers, which are listed for graphical reasons from bottom to top.
  • the uppermost layer of the layer system 1 shown is a film as a through-layer 1 .1 transparent to an excitation radiation 3 used (see FIG. 5) as well as to a radiation 6 of the degradation products of biological molecules caused by the excitation radiation 3 (see also FIG. 5) is.
  • a first film layer 1 .2 which is formed by aqueous condensate on the underside of the passage layer 1 .1.
  • a gas layer 1 .6 a second film layer 1 .3 and a food 1 .5.
  • the measuring methods of impedance spectroscopy and fluorescence spectroscopy and their applicability for measurement in the various layers of the layer system 1 are shown schematically. Measurements on the through-layer 1 .1 and the first film layer 1 .1 are possible as short-range measurements, measurements on the second film layer 1 .3 and the food 1 .5 are carried out as long-range measurements.
  • the gas layer 1 .6 is not measured directly by any of the methods given, but forms the boundary between near and far range measurements. The Gas layer 1 .6 exerts an influence on the corresponding methods, which is symbolized by the dashed line.
  • a layer system 1 is shown, that below the foodstuff 1 .5 a third film layer 1 .4, a suction pad (eg fleece insert) as absorbent layer 1 .8 and a non-transparent shell made of plastic as bottom layer 1 having.
  • electrodes 8 are arranged on the bottom layer 1 .7, impedance spectroscopic measurements can be carried out on the bottom layer 1 .7, the absorbent layer 1 .8 and the third film layer 1 .4. In further embodiments, the absorbent layer 1 .8 may also be omitted.
  • a first arrangement of excitation sources 2, electrodes 8 and a detection unit 7 is shown in FIG.
  • the detection unit 7 is surrounded by a UV source as a first excitation source 2.1 annular.
  • Two further, individually arranged second excitation sources 2.2 are arranged on each side of the detection unit 7 and the first excitation source 2.1.
  • an electrode 8 is present in each case.
  • the first and the second excitation sources 2.1 and 2.2 each emit radiation having a wavelength of 365 nm.
  • the detection unit 7 is designed for detecting electromagnetic waves having a wavelength in a range of 485 + 5 nm.
  • the second excitation sources 2.2 are provided for excitation of biological molecules in layers of a layer system 1 (see FIG. 5), while the first excitation source 2.1 is intended to produce excitations in the near field.
  • Fig. 5 provides a side view of the arrangement of Fig. 4 and also shows in a measuring space 14, a layer system 1 comprising a passage layer 1 .1, a first film layer 1 .2, a gas layer 1 .6, a second film layer 1 .3 , a food 1 .5, a third film layer 1 .4, a suction layer 1 .8 and a bottom layer 1 .7 and two additional electrodes 8 on the passage layer 1 .1.
  • the first excitation source 2.1 is shown in section.
  • An intensity and distribution of the excitation radiation of the first excitation source 2.1 is selected such that an excitation area 4 is effected in the first film layer 1 .2 in which the Excitation radiation 3 of the first excitation source 2.1 is sufficient to excite in the first film layer 1 .2 existing biological molecules for emitting a fluorescence radiation as a radiation 6.
  • the detection unit 7 is arranged vertically above the layer system 1 and a beam path 7.1 of the detection unit 7 is directed perpendicular to the layer system 1.
  • the beam path 7.1 runs through the excitation area 4 and intersects there the beam paths 2.1 1 of the first excitation source 2.1.
  • the beam paths 2.21 of the second excitation sources 2.2 are directed onto the second film layer 1 .3 and intersect the beam path 7.1 of the detection unit 7 in the second film layer 1 .3.
  • a far-range measurement of the second film layer 1 .3 is possible.
  • On the outer sides of the passage layer 1 .1 and the bottom layer 1 .7 four electrodes 8 are arranged in each case. These electrodes on each of the layers are each selectively interconnectable to allow impedance spectroscopic measurements in a flexible manner.
  • narrower and further electrodes 8 are placed in pairs on the upper side of the layer system 1. These serve for short-range measurement of the layer system 1. By simply pressing on the passage layer 1 .1 until contact with the food 1 .5 or by shaking or turning over the packaged foodstuff 1 .5, the first film layer 1 .2 on the underside of the passage layer 1 .1 amplified or even generated become.
  • the control of the device is performed by a computing unit 9.
  • FIG. 5 shows the exemplary embodiment in which the second outer layer 1 .3 and the surface of the foodstuff 1 .5 are irradiated for a short time by the further outer excitation sources 2.2 with a wide opening angle in the form of LEDs with UV light in order to generate fluorescence there , which can be detected via the detection unit 7. Since the first film layer 1 .2 is not irradiated directly below the detection unit 7, so only the far-range measurement of fluorescence at the second film layer 1 .3 is detected. If only the further inner first excitation source 2.1, which is also realized as an LED UV source, is activated, only becomes the first film layer 1 .2 below the detection unit 7 and less irradiated the more distant layers.
  • the further inner first excitation source 2.1 which is also realized as an LED UV source
  • the near range measurement of fluorescence can be performed.
  • both near and far range measurements can be measured in parallel as being amplifying or superimposing, if all excitation sources 2 are activated. Also temporally successive measurements are possible.
  • Complement to fluorescence spectroscopy, impedance spectroscopy can be performed.
  • the first excitation source 2.1 is formed of a UV source, a source of red light, a source of blue light, and a source of green light.
  • excitation regions 4 are formed in the first film layer 1 .2 and the second film layer 1 .3.
  • a region is highlighted in which an impedance spectroscopic short-range measurement can be performed by the electrodes 8 on the bottom layer 1 .7.
  • the so-called triplemode allows a differential assessment of the homogeneity and intensity of fluorescent degradation products, providing information about the current state of the food 1 .5.
  • This also includes the complementary impedance spectrometric bottom side (measurement in the vicinity of the bottom layer) effect. If the near-field measurements at the through-layer 1 .1 and bottom side measurement phase shift and the near and far-range measurements of fluorescence do not form a typical congruence, there is an indication that the food 1 .5 was repackaged, cleaned or chemically treated or the labeling changed.
  • the imprinted shelf life should not exceed 16 days in the case of unfrozen foodstuffs 1 .5. By recalculation and measurement this date is verifiable.
  • the second excitation sources 2.2 can be switched off.
  • the fluorescence is detected and measured in the detector unit 7.
  • Optical light interfaces in the device thereby couple directly or via light guides and possibly via photomultipliers and notch filters to a photodetector in the form of a photodiode, CCDs, spectrometer or colorimeter.
  • UV emission is coupled to the resting of the electrodes 8 for the impedance spectroscopy at the on the layer system 1.
  • the energy of the light is used to perform certain movements of the respective substance-specific molecule (case 1) or to stimulate electronic energy levels of the bonds and the atoms of these molecules (case 2).
  • the energy is converted into kinetic energy.
  • electrons are raised to higher energy levels.
  • the radiated energy is again indicated as an electromagnetic wave.
  • the energy (frequency, wavelength or wavenumber) of the radiated electromagnetic wave is the same as the injected electromagnetic wave.
  • the molecule loses radiation without energy.
  • the re-emitted light now has less energy than the incident light.
  • Such an effect is z.
  • the frequency of the fluorescent light is assigned exactly to a defined quantum mechanical electronic energy level transition and thus substance-specific or to a first approximation specific for a group of substances.
  • the frequency of the excitation light must be higher than the frequency of the fluorescent light. If UV light is irradiated onto a chemical compound of the Ptomaine type, many Ptomaine show, to a first approximation, a characteristic fluorescence with a wavelength of 485 nm.
  • the visible light covers the wavelength range from 700 to 400 nm.
  • the wavelength of the irradiated UV light can be at 380, 365, 320 or 254 nm.
  • the substances will fluoresce in any case. However, there is an optimum in which all other effects induced by an electromagnetic wave are small.
  • Ptomaine are u. a amines such.
  • cadaverine (1, 5-diaminopentane), which arise through the biochemical degradation of proteins and amino acids, inter alia, by decarboxylation.
  • the functional amino group (... C-NH 2 ) is characteristic of this group of substances.
  • the fluorescence at 485 nm results from the excitation of characteristic electronic quantum transitions in the chemical bond region of this functional group.
  • UV light having a wavelength of 365 nm is generated by means of a plurality of UV diodes (UV LEDs).
  • the UV light is guided centrally on the passage layer 1 .1 and the first film layer 1 .2 and centered on the food 1 .5.
  • the centrically annular excitation radiation 3 passes through the passage layer 1 .1 and the first film layer 1 .2.
  • Ptomaine are excited to fluorescence in the condensate of the first film layer 1 .2 (near zone).
  • the fluorescent light is conducted and evaluated as radiation 6 by a coaxial optical waveguide (in the interior of the annularly guided first excitation source 2.1) onto the detection unit 7.
  • the input of the light guide "looks" in the direction of the centrically guided light, and the output ends in the detection unit 7.
  • the acentric light reaches the food 1 .5 or the second film layer 1 .3 and stimulates the existing there ptomaine for fluorescence.
  • the fluorescent light can also be "seen” by the abovementioned coaxial light guide as radiation 6.
  • the detection unit 7 evaluates only a certain very narrow wavelength range (485 +/- 5 nm) of the light. Excessive visible light (radiation 6) and the exciting UV light as excitation radiation 3 are hidden by the Cutt-Off filter.
  • Fig. 6 and 7 the excitation of fluorescence is as described above.
  • s red-green-blue semiconductor light-emitting
  • s red-green-blue semiconductor light-emitting
  • s red-green-blue semiconductor light-emitting
  • s red-green-blue semiconductor light-emitting
  • s red-green-blue semiconductor light-emitting
  • RGB LEDs provided as first excitation sources 2.1
  • a spectroscopic and / or colorimetric unit as a detection unit.
  • the peculiarity of colorimetry (colorimetry) as an alternative and / or supplement to classical fluorescence spectroscopy by means of spectrometers or wavelength-optimized photodiodes with notch filters is, in addition to the respective space, cost and information technology effort, the conversion of the spectral measured values of the colorimeter to the desired color coordinates.
  • a process-related high computational effort can be outsourced to external servers.
  • the standard spectral color values defined by the International Commission on Illumination (CIE) have prevailed. These basic numbers are tabulated at a distance of one nanometer.
  • CIE International Commission on Illumination
  • this "influenced" spectral radiance For the color stimulus that hits the eye, the color in the true sense of the word, this "influenced" spectral radiance must be used, either the spectral reflectance curve for surface colors or the spectral transmission curve for supervisory colors
  • the selected coefficients for remission and transmittance are determined and added here, and the radiation distribution of the light source can be summarized and measured in this spectral interval, and the color values are determined by measuring the color stimuli in these intervals.
  • the remission of the incident light is also changed by the surface of the layers. If fluorescence effects are added, the spectral components below the fluorescent color are absorbed.
  • bottom layer 1 .7 can vary from light to deep black
  • the arrangement shown in FIGS. 6 and 7 is advantageous for colorimetry.
  • FIG. 8 A schematic overview of the individual method steps and some device elements is given in FIG. 8.
  • a portable measuring device 1 1 for carrying out the method is shown in simplified form in FIG. 9.
  • the measuring device 1 1 contains an excitation source 2 and the detection unit 7.
  • a receiving tray 1 1 .1 is formed, which serves to accommodate a telecommunications device 12.
  • a connection to the power supply and data linkage of meter 1 1 and telecommunication device 12 is realized via an adapter 1 1 .2 of the measuring device 1 1.
  • the meter 1 1 via a program installed on the meter 1 1 program can be operated.
  • communication with a virtual memory and arithmetic unit 10 is possible via the telecommunication device 12, via which part of the computing effort and the memory capacity required for carrying out the method according to the invention is provided.
  • This communication can be done by the telecommunication device 12 via the usual telecommunications channels.
  • the meter 1 1 In the right half of the application of the meter 1 1 is shown connected to telecommunications device 12 by a user at a point of sale of packaged food.
  • the device connected to the telecommunication device 12 1 1 on a layer system to be examined 1, here a packaged in foil piece of meat, pressed and controlled by the telecommunication device 12, the meter 12.
  • the meter 12 Depending on the specific configuration of the measuring device 11, at least one fluorescence spectroscopic and / or colorimetric measurement of the first film layer 1 .2 of the layer system 1 is carried out.
  • the detected radiation 6 of the excited degradation products is evaluated, a concentration of the degradation products is determined and compared with a permissible limit value.
  • a display 13 is displayed, which signals a color code undershooting or exceeding the limit by green and red fields of the display 13. During the measurement or in case of ambiguous results of the measurement, a yellow field is displayed.
  • the measuring device 1 1 can emit a signal tone.
  • an impedance spectroscopic measurement of the bottom layer 1 .7 can be carried out by the user, after which the measurements of both methods are examined by the measuring device 1 1 for congruence with one another. In the case of an impermissible deviation, an indication is sent to the user. At the same time, information can be sent to the competent food control authority.
  • FIG. A device according to the invention can be integrated in a cooling device or be available as a household appliance.
  • the device according to the invention and the invention also for testing and control purposes of example, consumer protection organizations or with the Monitoring of food commissioned authorities and be provided, for example, with a corresponding inspection certificate.
  • Fig. 1 1 shows possible partial steps of the process and their networking possibilities with each other.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Biotechnology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'invention concerne un procédé de détection de produits de dégradation de molécules biologiques dans des couches d'un système de couches (1), selon lequel les produits de dégradation sont exposés à un rayonnement d'excitation (3), un rayonnement (6) des produits de dégradation résultant du rayonnement d'excitation (3) est détecté, et une concentration des produits de dégradation est déterminée sur la base d'une comparaison des valeurs mesurées du rayonnement (6) avec des données de référence, le procédé étant caractérisé en ce que l'on prépare un système de couches (1) qui comprend une couche de transmission (1.1) transparente pour le rayonnement d'excitation (3) et pour le rayonnement (6), ainsi qu'au moins une autre couche réceptrice qui contient les produits de dégradation et en ce que le rayonnement d'excitation (3) est introduit dans la couche réceptrice à travers la couche de transmission (1.1). L'invention concerne en outre un dispositif ainsi qu'un appareil de mesure portatif (11) pour la détection des produits de dégradation.
EP13740203.8A 2012-06-18 2013-06-18 Procédé, dispositif et appareil de mesure portatif pour la détection de produits de dégradation de molécules biologiques dans des couches d'un système de couches Withdrawn EP2861973A1 (fr)

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Application Number Priority Date Filing Date Title
DE102012105291A DE102012105291A1 (de) 2012-06-18 2012-06-18 Verfahren, Vorrichtung und tragbares Messgerät zur Detektion biologischer Moleküle in Schichten eines Schichtsystems
PCT/DE2013/100220 WO2013189488A1 (fr) 2012-06-18 2013-06-18 Procédé, dispositif et appareil de mesure portatif pour la détection de produits de dégradation de molécules biologiques dans des couches d'un système de couches

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WO2013189488A1 (fr) 2013-12-27
US20150192521A1 (en) 2015-07-09

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