WO2024074271A1 - Procédé de concentration d'au moins une substance cible anthropogène dans un liquide échantillon - Google Patents

Procédé de concentration d'au moins une substance cible anthropogène dans un liquide échantillon Download PDF

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Publication number
WO2024074271A1
WO2024074271A1 PCT/EP2023/075071 EP2023075071W WO2024074271A1 WO 2024074271 A1 WO2024074271 A1 WO 2024074271A1 EP 2023075071 W EP2023075071 W EP 2023075071W WO 2024074271 A1 WO2024074271 A1 WO 2024074271A1
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WO
WIPO (PCT)
Prior art keywords
sample
liquid
superabsorbent
target substance
anthropogenic
Prior art date
Application number
PCT/EP2023/075071
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German (de)
English (en)
Inventor
Timo Hillebrand
Elmara Graser
Wiebke JACOBI
Original Assignee
Ist Innuscreen Gmbh
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
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Application filed by Ist Innuscreen Gmbh filed Critical Ist Innuscreen Gmbh
Publication of WO2024074271A1 publication Critical patent/WO2024074271A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/405—Concentrating samples by adsorption or absorption
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/06—Investigating concentration of particle suspensions
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/06—Investigating concentration of particle suspensions
    • G01N15/075—Investigating concentration of particle suspensions by optical means
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
    • G01N2001/4088—Concentrating samples by other techniques involving separation of suspended solids filtration
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N2015/0042—Investigating dispersion of solids
    • G01N2015/0053—Investigating dispersion of solids in liquids, e.g. trouble

Definitions

  • the invention relates to a method for concentrating at least one anthropogenic target substance in a sample liquid, which anthropogenic target substance consists of particles and/or particles with an average particle size in the nanometer range.
  • Nanoparticles and nano particles are playing an increasingly important role in our lives. This applies, for example, to the use and, in this context, the production of these particles and particles for a variety of applications in the pharmaceutical industry (for example in connection with encapsulated mRNA active ingredients), food technology or the electrical engineering or electronics industry (for example in connection with quantum dots).
  • Nanoparticles can be used to transport drugs to desired organs, as they can penetrate the blood-brain barrier (e.g. suitable for use in the pharmaceutical industry) and the skin (e.g. suitable for use in the cosmetics industry). Nanoparticles are also used for diagnostic applications, for example for color detection in rapid tests based on colloidal gold. They also stabilize food and thus ensure longer shelf lives.
  • blood-brain barrier e.g. suitable for use in the pharmaceutical industry
  • the skin e.g. suitable for use in the cosmetics industry.
  • diagnostic applications for example for color detection in rapid tests based on colloidal gold. They also stabilize food and thus ensure longer shelf lives.
  • Nanoparticles are almost exclusively anthropogenic substances. This means that the nanoparticles have been produced by humans through industrial, commercial and municipal processes and not, for example, by biological organisms. Another aspect of nanoparticles is the pollution of our planet, as nanoparticles are often difficult to biodegrade. Nanoparticles are created, for example, from waste from the plastics industry or from the partial decomposition of plastic waste in nature (so-called secondary nanoplastics). The worldwide pollution of the environment with waste from the plastics industry is a global problem. Nanoplastic waste with particle sizes of less than 1 pm enters the ecosystem unhindered and is often not stopped by sewage treatment plants. The damage caused by microplastics and nanoplastics includes the accumulation of particles in the tissue of aquatic creatures.
  • the plastic components ⁇ 0.33 mm i.e. parts of the nanoparticles and nanoparticles in the micrometer range and all nanoparticles and nanoparticles in the nanometer range
  • the plastic components ⁇ 0.33 mm i.e. parts of the nanoparticles and nanoparticles in the micrometer range and all nanoparticles and nanoparticles in the nanometer range
  • Micro- and nanoplastic waste is detected using spectroscopic methods (e.g. Raman or infrared spectroscopy), thermoanalytical methods (e.g. dynamic differential scanning calorimetry (DSC)), gas chromatograph mass spectrometer (GC/MS), microscopy (optical, fluorescence-based) or fluorescence-based flow cytometry.
  • spectroscopic methods e.g. Raman or infrared spectroscopy
  • thermoanalytical methods e.g. dynamic differential scanning calorimetry (DSC)
  • GC/MS gas chromatograph mass spectrometer
  • microscopy optical, fluorescence-based
  • the object of the invention is to provide a simple, rapid and universally applicable method for concentrating at least one anthropogenic target substance in a sample liquid.
  • the solution is based on the use of so-called superabsorbents, with which any aqueous sample liquid can be processed in order to concentrate the nanoparticles contained in the sample liquid.
  • Superabsorbent polymers are plastics that are able to absorb many times their own weight in polar liquids. These are mainly water or aqueous solutions. When the liquid is absorbed, the superabsorbent swells and forms a hydrogel. Hydrogels can form all cross-linked polymers that are polar (e.g. polyacrylamide, polyvinylpyrrolidone, amylopectin, gelatin, cellulose). However, a copolymer of acrylic acid is usually used.
  • the product is traditionally used, for example, as white granules with particle sizes of 100 to 1000 pm. It is mainly used in baby diapers, sanitary napkins, incontinence care, in dressing material and, in small quantities, in cable sheathing for deep-sea cables.
  • Other areas of application include so-called gel beds, gel-forming extinguishing agents in fire fighting, as a mechanical stabilizer for cut flowers in a vase or as an additive for plant soil to permanently store water.
  • acrylic acid neutralized with potassium hydroxide is used because of its better environmental compatibility.
  • the use of superabsorbents is known as toys under names such as "water beads", “aqua beads” or “water beads”. These are superabsorbents which are commercially available in the form of balls of variable size (submillimeters to centimeters).
  • the invention was based on the following unexpected observation: A water sample was mixed with fluorescent nanoparticles with an average particle size of 30 nm. After adding commercially available water pearl beads and an incubation period in which the beads swelled to several times their original volume, it was found that the nanoparticles were not absorbed by the superabsorbents, but were concentrated in the remaining liquid.
  • the method according to the invention for concentrating at least one anthropogenic target substance in a sample liquid comprises:
  • “Anthropogenic substances” are substances that are not created by nature but by humans, for example through industrial, commercial or municipal processes. They include, for example, plastics, but also pesticides, pharmaceuticals, personal care products and industrial chemicals as well as their degradation products and metabolites. Biomolecules that are created by microorganisms (e.g. enzymes, DNA/RNA fragments, etc.) and have similar sizes in the nanometer range do not fall under the particles or particles of anthropogenic substances mentioned in this application.
  • microorganisms e.g. enzymes, DNA/RNA fragments, etc.
  • the first sample taken from the liquid portion of the mixture of liquid and superabsorbent present after incubation can be the entire remaining liquid portion.
  • the first sample can be used immediately for subsequent analysis, e.g. using a spectroscopic method. It can also be further concentrated in a cascading process in one or more additional stages, e.g. by adding a superabsorbent again or adding it to a superabsorbent again and incubating again, or by using a conventional method for concentrating target substances, e.g. using one of the methods given in the introduction. If only part of the liquid portion is taken as the first sample, the target substance in the liquid portion of the mixture remaining after sampling can be further concentrated by incubating again for a second period. Both variants of the process can be repeated several times so that a higher concentration of the target substance is obtained at each stage of the cascaded concentration.
  • the method may comprise a further concentration of the target substance in the first sample taken after the first sample has been taken.
  • Further concentration of the target substance in the first sample taken can be carried out by means of a filtration, ultrafiltration or precipitation reaction technique.
  • the further concentration of the target substance in the first sample taken can also be carried out again - and optionally repeated in cascading fashion once or several times - by carrying out the following process steps:
  • the concentrated first sample taken from the liquid portion remaining after incubation can comprise the entire volume of the liquid portion.
  • the concentrated first sample can be a partial volume of the remaining liquid portion.
  • the target substance can be further concentrated in the concentrated first sample or in a further concentrated first sample obtained by further concentration, in particular by means of a superabsorbent, by means of a filtration, ultrafiltration or precipitation reaction. This is advantageous if the volume of the concentrated first sample corresponds to only a few milliliters, e.g. 1 to 10 ml.
  • the method according to the invention for concentrating the target substance in the liquid portion of the mixture remaining after taking the first sample can comprise the following further steps: renewed - and optionally cascading one or more times repeated - concentration of the target substance in the liquid portion of the mixture remaining after taking the first sample by - re-incubating the mixture of the remaining liquid portion and the superabsorbent remaining after the first sample has been taken for a third period; and
  • the concentration of the target substance is determined, among other things, by the predeterminable duration of the third period and the conditions prevailing during incubation.
  • the initially used liquid volume can contain a polar liquid, in particular as the main component.
  • the liquid volume can contain a polar solvent, in particular as the main component.
  • the liquid volume can consist of a polar solvent to a mass fraction of at least 50%.
  • the polar liquid or the polar solvent can be water, for example.
  • the target substance is a nanoparticle or nano particles.
  • a substance consists of an anthropogenic substance, in particular of at least one natural polymer, of at least one biocompatible synthetic polymer, of an inorganic material or of an organic material.
  • the superabsorbent can be a plastic or comprise a plastic that absorbs a portion of the liquid volume, e.g. a polar solvent contained in the liquid volume such as water, to form a gel or hydrogel.
  • the plastic is advantageously selected so that it absorbs essentially no nanoparticles. This is the case, for example, with the mentioned superabsorbents made from the mentioned polymer or copolymer materials, e.g. with the commercially available water pearls, water beads, etc.
  • the superabsorbent can be used in the form of particles, e.g. as a powder, as granules or in the form of geometric bodies, in particular spheres (spherical particles). It can thus be added to the liquid volume or the first sample in the form of such particles or the liquid volume or the first sample can be added to the superabsorbent in this form.
  • the particles or spheres can have a diameter of between 100 and 5000 pm.
  • the superabsorbent is commercially available superabsorbent beads, for example superabsorbent beads available under the names “Aquabeads”, “Water Beads”, “Water Pearls”, “Aqua Beads”, “Hydro Beads”, “Gel Beads”.
  • the volume of the liquid portion remaining after the incubation step, and thus the concentration of the target substance in the remaining liquid portion can be controlled by the length of the incubation period or periods, by the size and number of superabsorbent particles or superabsorbent spheres added to the initially used liquid volume of the sample liquid or the first sample, and/or by the temperature prevailing during incubation.
  • the sample liquid may be a filtrate or a centrifugation supernatant. This means that one or more separation steps are carried out before concentration in order to remove further components from the sample.
  • the sample liquid is an environmental sample.
  • An environmental sample is taken from the environment, for example from bodies of water such as lakes or rivers. But sediment or gas samples, for example, which are then mixed with a liquid, are also referred to as environmental samples.
  • the invention further comprises a method for detecting an anthropogenic target substance in a sample liquid, comprising:
  • the physical detection method is used to determine whether certain nanoparticles or nano particles are present in the concentrated sample and how high their concentration is. In the initial liquid volume, these would not be detectable using the physical detection method because their concentration is too low.
  • a microscope is used for qualitative and/or quantitative determination.
  • the nanoparticles or nano particles are counted in a section of a defined size.
  • a fluorescence measuring method or a spectroscopic measuring method is used. For this, the nanoparticles or nano particles must be
  • Nanoparticles contain a fluorescent dye, which is added in particular before concentration.
  • the invention also includes the use of a superabsorbent for the one-time or multiple cascading concentration of the anthropogenic target substance in a liquid sample.
  • the liquid sample can contain a polar liquid, in particular water, wherein the superabsorbent is designed to absorb the polar liquid, e.g. water, or at least a portion of the polar liquid, to form a hydrogel.
  • the superabsorbent can be formed from the materials described above and in the previously described Embodiments as granules or, particularly preferably, in the form of beads, in particular commercially available water beads, water beads or aqua beads, can be used.
  • the cascaded concentration can comprise several stages. After each stage, a sample can be taken for analysis and the concentration process continued, or the sample or part of the sample concentrated in at least one stage after incubation of the sample with the superabsorbent can subsequently be further concentrated using other known methods.
  • the invention also includes a kit for carrying out the method described above.
  • the kit can, for example, include one or more containers with superabsorbent material, into which a user can then add an initial volume of liquid for concentration or a sample that has already been concentrated in a first stage.
  • the liquid concentrated according to the described method or according to the described use i.e. the liquid portion present after incubation, or a sample taken from the concentrated liquid can be fed manually or automatically to a laboratory device for further treatment or analysis.
  • a liquid or sample of the liquid can be fed into a cartridge, in particular a microfluidic cartridge, of an automatic analysis device for the automated implementation of a detection of the target substance by means of a physical detection method for the qualitative and/or quantitative determination of the anthropogenic target substance. This can be done manually or automatically.
  • Fig. 1 is a schematic representation of the cascading concentration of a target substance in a liquid: a) liquid before adding a superabsorbent; b) liquid after adding a superabsorbent and incubating the mixture; c) first sample taken from the mixture after adding another superabsorbent and incubating the mixture; d) remaining mixture of liquid and superabsorbent, if applicable after taking the first sample and after incubating again;
  • Fig. 2 is a representation of samples and blank samples, some of which were obtained by means of the concentration according to the invention and exposed to UV light; and Fig. 3 Evaluation of the measurement data: a) a graphical representation of the mean values of the measured values in a bar chart; b) a correlation between the degree of concentration and the increase in fluorescence of the samples.
  • Further processing can, for example, be a quantitative and/or qualitative detection of the target substance in the liquid.
  • the detection is carried out using a spectroscopic or microscopic method, for example.
  • the degree of concentration and the speed of this process can be controlled very precisely by the type of superabsorbent used, by the amount used, or by the incubation time and/or the incubation temperature.
  • the method can therefore easily solve the problem of processing low-concentration samples for further processing of the target substances of interest, in particular nanoparticles, or their detection.
  • the method shown in Fig. 1 a and b does not require equipment such as ultracentrifuges, expensive ultrafiltration membranes, complex processes such as PEG precipitation or general precipitation reactions for concentrating nucleic acids, etc.
  • the method can be used universally with regard to the type of nanoparticles.
  • a further advantage is that the superabsorbents are non-toxic and harmless and often biodegradable. The method according to the invention can therefore greatly simplify the examination of low-concentration nanoparticles.
  • a cascading concentration of the target substance is an option.
  • the described method with the above-mentioned steps 1-3 can be used to concentrate the target substance.
  • the volume of the first sample 4 can be reduced to reconcentrate the target substance. This can be done either by means of a conventional filtration or precipitation process or other conventional methods. Alternatively, the reconcentration of the target substance in the first sample 4 can also be done, as shown in Fig.
  • the latex nanoparticles were provided by the Fraunhofer Institute for Applied Polymer Research.
  • the concentration of the latex particles in the stock solution was 2.02 M%.
  • the particles were added to a 500 ml water sample, thereby producing a 1:10,000-fold dilution.
  • a superabsorbent in the form of commercially available “water beads” was added to the water sample.
  • samples PO, P1, P2 were taken at different times, each of which corresponded to a different concentration.
  • the samples are divided into blank samples L0, L1, L2 for control (without latex nanoparticles) and samples PO, P1, P2, which contain the latex nanoparticles.
  • the samples are as follows: Blank sample L0: ultrapure water without latex particles (500 ml) Blank sample L1: concentration of 500 ml ultrapure water to 40 ml ultrapure water Blank sample L2: further concentration of L 1 to 1 ml ultrapure water Sample PO: 1:10,000 dilution of the latex particle stock solution in 500 ml ultrapure water Sample P1: concentration of 500 ml ultrapure water to 40 ml ultrapure water Sample P2: further concentration of P 1 to 1 ml ultrapure water The particles in the respective samples were detected by measuring the fluorescence of the dye contained in the latex nanoparticles.
  • Fig. 1 shows a qualitative detection of the latex nanoparticles.
  • the respective samples PO, P1, P2 and the blank samples LO, L1, L2 were transferred in triplicate to a UV-transparent measuring plate. The measuring plate with the samples was then exposed to UV light. Exposure to UV light stimulates fluorescence in the samples that contain latex particles. No fluorescence can be seen in the blank samples LO, L1, L2 and an increasing fluorescence with increasing concentration in the samples PO, P1, P2, which contain the latex nanoparticles.
  • Table 1 shows a quantitative measurement of the respective samples using a fluorescence measuring device. The measurements are carried out by exciting the samples PO, P1, P2 and the blank samples LO, L1, L2 (each in triplicate) at an emission of 559 nm. While the values of the blank samples LO, L1, L2 remain stable and low with concentration, the values of the samples PO, P1, P2 that contain the nanoparticles increase proportionally to the degree of concentration.
  • Fig. 2 the measured values listed in Table 1 are graphically displayed or evaluated.
  • Fig. 2a the mean values of the respective measured values of a sample are shown as a bar chart.
  • Fig. 2b a correlation is shown between the degree of concentration and the increase in fluorescence of the samples PO ("1"), P1 ("2") and P2 ("3"), which contained nanoparticles.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

L'invention concerne une concentration d'au moins une substance cible anthropogène dans un liquide échantillon, laquelle substance cible anthropogène est constituée de particules et/ou de matières particulaires ayant une taille moyenne de particule ou de matière particulaire dans la plage nanométrique, comprenant : - l'ajout d'un superabsorbeur (2) à un volume initial de liquide (1) du liquide échantillon ou l'ajout du volume de liquide (1) au superabsorbeur (2), - l'incubation, pendant une première période (t1), du mélange obtenu par mélange du superabsorbeur (2) et du volume de liquide (1), et - l'élimination d'un premier échantillon (4) de la partie liquide (3) du mélange présent après incubation.
PCT/EP2023/075071 2022-10-06 2023-09-13 Procédé de concentration d'au moins une substance cible anthropogène dans un liquide échantillon WO2024074271A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102022125783.5 2022-10-06
DE102022125783.5A DE102022125783A1 (de) 2022-10-06 2022-10-06 Verfahren zur Aufkonzentrierung mindestens einer anthropogenen Zielsubstanz in einer Probenflüssigkeit

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WO2024074271A1 true WO2024074271A1 (fr) 2024-04-11

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Citations (4)

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US20070111194A1 (en) * 2003-12-15 2007-05-17 Preentec Ag Method for the concentration and purification of biological compounds
US20130231460A1 (en) * 2010-09-08 2013-09-05 Qiagen Gmbh Method and device for concentrating target compounds
EP2283026B1 (fr) 2008-05-08 2017-11-01 AJ Innuscreen GmbH Procédé permettant d enrichir et d isoler des biomolécules ou des virus
WO2021065300A1 (fr) * 2019-09-30 2021-04-08 富士フイルム株式会社 Méthode d'essai immunologique et gabarit de condensation

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DE102017220514A1 (de) 2017-11-16 2019-05-16 Unisensor Sensorsysteme Gmbh Verfahren und Vorrichtung zur Detektion von Fremdstoffen in einem flüssigen Medium
EP3759472B1 (fr) 2018-02-26 2023-01-18 Basf Se Procédé de mesure de la perméabilité d'un superabsorbant
CN114174797A (zh) 2019-07-04 2022-03-11 巴斯夫欧洲公司 测定超吸收剂特性的方法
WO2023089078A1 (fr) 2021-11-22 2023-05-25 Ist Innuscreen Gmbh Procédé de concentration en cascade d'au moins une substance cible dans un échantillon liquide
WO2023088991A1 (fr) 2021-11-22 2023-05-25 Ist Innuscreen Gmbh Procédé de concentration d'au moins une substance cible dans un échantillon liquide
DE102021134613A1 (de) 2021-12-23 2023-06-29 Ist Innuscreen Gmbh Verfahren zum Aufkonzentrieren mindestens einer biologischen Zielsubstanz in einer Probeflüssigkeit und Verfahren zur automatisierten Online-Detektion mindestens einer biologischen Zielsubstanz in einer Probeflüssigkeit

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US20070111194A1 (en) * 2003-12-15 2007-05-17 Preentec Ag Method for the concentration and purification of biological compounds
EP2283026B1 (fr) 2008-05-08 2017-11-01 AJ Innuscreen GmbH Procédé permettant d enrichir et d isoler des biomolécules ou des virus
US20130231460A1 (en) * 2010-09-08 2013-09-05 Qiagen Gmbh Method and device for concentrating target compounds
WO2021065300A1 (fr) * 2019-09-30 2021-04-08 富士フイルム株式会社 Méthode d'essai immunologique et gabarit de condensation

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