WO2015066459A1 - Dispositif de détection d'analyte et procédés d'utilisation de celui-ci - Google Patents

Dispositif de détection d'analyte et procédés d'utilisation de celui-ci Download PDF

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Publication number
WO2015066459A1
WO2015066459A1 PCT/US2014/063398 US2014063398W WO2015066459A1 WO 2015066459 A1 WO2015066459 A1 WO 2015066459A1 US 2014063398 W US2014063398 W US 2014063398W WO 2015066459 A1 WO2015066459 A1 WO 2015066459A1
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solution
subject
color
polymer
skin
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PCT/US2014/063398
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Richard KNOP
Tiffany GRIFFIN
Victor PEREZ-LUNA
William B. LESLIE
Edward Johnson
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Medtech Bioscience, Llc
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Publication of WO2015066459A1 publication Critical patent/WO2015066459A1/fr

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
    • A61B10/0045Devices for taking samples of body liquids
    • A61B10/0064Devices for taking samples of body liquids for taking sweat or sebum samples
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
    • A61B2010/0003Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements including means for analysis by an unskilled person
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
    • A61B2010/0009Testing for drug or alcohol abuse

Definitions

  • the present invention generally relates to the field of analyte detection.
  • this invention is directed to devices and related methods for indicating the presence of a target analyte in a bodily secretion located on the skin of a subject.
  • the target analyte is preferably an illicit drug or metabolite thereof.
  • Analyte detection is commonly used in the medical field to diagnose disease or detect drugs in toxicology. There are many modalities that can be used and a wide variety of biological specimens that can be tested.
  • Passive testing usually involves collecting a sample to send out to a laboratory for a more in-depth analysis.
  • Active testing usually involves immediate analyses and has been called "quick testing.”
  • Sweat is a unique medium for biological analyte testing. Moisture is lost through the skin via two pathways. The first pathway is insensible sweat and the diffusion of moisture through the dermal and epithelial layers. The second pathway is called sensible sweat. This sweat comes from apocrine, eccrine and sebaceous glands. There have been several variations of sweat collection patches developed over the years to collect this fluid. Details of the history of these patches and more specific information on sweat testing can be found in Forensic Science and Medicine: Drug Testing in Alternate Biological Specimens Edited by: A. J. Jenkins (Humana Press; 1st edition (2008 ⁇ .
  • Sweat testing allows for the detection of the compound of interest in addition to metabolites. It also can avoid issues with urine drug testing such as dilution, specimen adulteration and substitution. Sweat collection patches can be used to collect a sample and then send it out to a laboratory for further analysis. The benefits and details of the development of sweat collection patches can be found extensively described in, e.g., United States Patent 6,443,892 by Kidwell.
  • U.S. Patent 6,443,892 is exemplary of sweat collection patches that are passive sensors. It is worn for approximately a week and then sent to a laboratory for further analysis. The process of collection and analysis can take several weeks. This can be advantageous over urine and blood drug testing, as some analytes will only be detectable for small windows of 24-48 hours within that one-week period. To gain comparable results of a patch, one would need to conduct daily urine testing. The long time frame is problematic in rehabilitation, criminal, medical and employment settings. A patient in a rehabilitation program or an employee operating dangerous equipment on an illicit substance needs to be identified immediately.
  • Sweat patches such as the device described in U.S. Patent 6,443,892 may include a security feature.
  • This feature is normally a specific identification number printed on the test patch. They may also contain an adhesive with a security feature to show tampering.
  • the tape may change colors if it is exposed to air or not re-adhered to the skin. This does not prevent a patient from easily disposing of a test patch if they are concerned with an adverse result.
  • a sweat patch can collect both the analyte of interest in addition to metabolites. This can be important since certain compounds will break down to the same metabolite. For example, codeine and heroin produce the same metabolites in the urine. To properly identify which substance a patient ingested, a sweat patch may be preferred over a urine test.
  • Prior sweat patches typically have three main components: 1 ⁇ a polyurethane/adhesive layer such as a transparent wound dressing that acts as both an adhesive and membrane 2] a release liner that is a medical grade tissue paper that the test pad rests on to prevent the test pad from touching the adhesive membrane and 3 ⁇ a collection pad that can be made of medical grade cellulose or equivalent material.
  • the standard method to use a sweat patch is to first swab an area of the arm with an alcohol wipe or equivalent cleaning wipe. To allow any volatile cleaning fluid to evaporate and to place the sweat collection patch firmly on the test site. The sweat collection patch is then usually left attached for a minimum of 24 hours or as long as a week. The patch is then removed. The pad is transferred using sterile technique to a transport container. The container is then mailed to a diagnostic laboratory for analysis by, e.g., gas chromatography/mass spectrometry (GC/MS] or liquid chromatography/mass spectrometry (LC/MS] testing. Gas chromatography-mass spectrometry (GC/MS] is a common method of drug testing used to confirm a positive drug test.
  • GC/MS gas chromatography/mass spectrometry
  • LC/MS liquid chromatography/mass spectrometry
  • GC/MS is used for the quantification and identification of analytes in complex mixtures.
  • Liquid chromatography-mass spectrometry is a complementary technique to GC/MS. Compounds that are difficult to analyze by GC/MS are contenders for LC/MS.
  • Active analyte testing can give immediate results.
  • the most common active analyte tests typically involve urine testing quick test strips, which use variations of the Enzyme-linked immunosorbent assay or ELISA test.
  • Quick analyte testing can be colorimetric, antibody based, use polymers, microfluidic chips or more novel nanotechnology based methodologies.
  • a common feature of many of these quick tests is that they produce a color change within the human visual spectrum and give an immediate result when placed into contact with a biological specimen.
  • a preliminary positive result with these tests is typically confirmed by sending the sample out to a laboratory for further analysis (U.S. Patent 8,071,394 is exemplary of such methodology].
  • Colorimetric assays can be used to display a large variety of information via visual indicators that result from chemical reactions. They can make use of chromophores or fluorophores. Colorimetric assays commonly utilize the absorbance of light. The wavelength of light the solute absorbs can be directly comparable to the concentration of the solute. Colorimetric assays are a simple and widely available method of establishing a variety of information. Colorimetric assays can help in differentiating one analyte from other analytes. They have the ability to quantify catalytic activity of substances. Colorimetric assays have the capacity to produce toxicological summaries; these assays can also be used in determining concentrations of substances.
  • Colorimetric assays have recently been used in the detection of GHB and other illicit drugs as shown in, e.g., U.S. Patent 6,713,306, U.S. Patent 7,238,533, U.S. published patent application 2008/0102482 and U.S. Patent 6,153,147, where test strips, coasters and even fingernail polish/paints have been the device of choice to host the assay.
  • Colorimetric testing for drugs of abuse also exists for urine sampling (e.g., see Forensic Sci Int. 2002 Apr 18; 126(2 ⁇ :114-7 ⁇ . Rapid colorimetric screening test for gamma-hydroxybutyric acid (liquid X] in human urine (Alston WC 2nd, Ng K. or the BUHLMANN enzymatic GHB assay ⁇ .
  • Enzyme-linked immunosorbent assay is a rapid, highly sensitive test that utilizes an enzyme as well as components of the immune system, an antibody or antigen, to detect various analytes.
  • ELISA testing methods differ depending on the immune system component being detected. Although testing methods differ, the last step in ELISA is the production or nonproduction of a colored product for positive or negative results, respectively.
  • ELISA has been used as a detection method for illicit drugs in many active quick test formats.
  • One of the most common forms of ELISA testing is a sandwich ELISA test.
  • wipe U.S. Patent 6,514,773 ⁇ that uses a modified immunoassay to gain an immediate test result to see if there is drug residue on a surface.
  • Such a wipe can be used to test the biological fluids of sweat or saliva. These tests may not act continuously but may be multi-chambered to test multiple analytes (see U.S. Patents: 6,881,378, 7,189,522, 7,267,992, 7,354,776, 7,682,801, and 7,879,597 ⁇ .
  • microfluidic chips One emerging class of sensors is microfluidic chips.
  • U.S. patent 7,931,592 shows an example of a microfluidic chip used to sample transdermal analytes and remotely monitor a patient. Different testing methods can be used within the chip. Continuous or non-continuous testing may be used. If non-continuous testing is used, a chip may control the timing of the fluidic intake into non-continuous test channels to allow for a quasi-continuous monitoring at regularly scheduled intervals.
  • microfluidic devices that use micro-needle sampling of bodily fluids at regular intervals as in U.S. Patent 7,344,499. Many of these microfluidic chips can also use colorimetric assays.
  • An integrated digital microfluidic lab-on-a-chip for clinical diagnostics on human physiological fluids is described at Srinivasan V, Pamula VK, Fair RB. Lab Chip. 2004 Aug; 4(4 ⁇ :310-5. Epub 2004 May 26.
  • the present invention provides a device for indicating the presence of a target analyte that is contained in a bodily secretion present on a subject's skin, comprising: (a] a testing pad to be positioned in contact with a subject's skin; (b] an external layer to be positioned opposite subject's skin; (c] a sensor means configured to provide a colorimetric indication of the presence of a target analyte in the subject's bodily secretion while the device is securely-fastened to the subject's skin; and (d] a securing means in which the testing pad, external layer, and sensor means are contained, said securing means configured to be securely-fastened to the subject's skin.
  • the bodily secretion is preferably the subject's sweat.
  • the target analyte is preferably an illicit drug or metabolite thereof, including, but not limited to, a narcotic, stimulant, depressant, hallucinogen, or cannabis.
  • the device is configured to securely- fasten around the wrist of a human subject.
  • the senor means comprises a molecularly- imprinted polymer material that facilitates the colorimetric indication of the presence of the target analyte in the subject's bodily secretion.
  • the senor means may comprise a nanocomposite material that facilitates the colorimetric indication of the presence of the target analyte in the subject's bodily secretion.
  • the senor means provides a colorimetric indication of the target analyte in the subject's bodily secretion that is in a portion of the electromagnetic spectrum visible to the human eye.
  • the device further includes a secretion impermeable layer positioned between the external layer and the test pad.
  • the device may, alternatively, further include a removable liner which is positioned in contact with the test pad, said removable liner configured for removal before the device is fastened to the subject's skin.
  • the device may further include a secretion wicking layer which is positioned in contact with the sensor means, said secretion wicking layer absorbent to the bodily secretion and in contact with the subject's skin when the device is fastened to the subject's skin.
  • the invention encompasses a method for directly- indicating the presence of a target analyte contained in a bodily secretion present on a subject's skin.
  • a method for directly- indicating the presence of a target analyte contained in a bodily secretion present on a subject's skin includes steps of: (a] securely fastening a device as described and claimed herein on a subject's skin; and (b] examining said device while it is securely-fastened to the subject's skin; wherein said sensor means provides a colorimetric indication of the presence of a target analyte in a bodily secretion present on the skin of the subject.
  • the method is carried out as part of an illicit drug rehabilitation program or illicit drug monitoring program.
  • the method is preferably practiced on a human subject, and the target analyte is an illicit drug or metabolite thereof, including, but not limited to, a narcotic, stimulant, depressant, hallucinogen, cannabis or a metabolite thereof.
  • the device used in the method may comprise a molecularly-imprinted polymer material that facilitates the colorimetric indication of the presence of the target analyte in the subject's bodily secretion.
  • the device may comprise a nanocomposite material that facilitates the colorimetric indication of the presence of the target analyte in the subject's bodily secretion. It is preferred that the device provides a colorimetric indication of the target analyte in the subject's bodily secretion that is in a portion of the electromagnetic spectrum visible to the human eye.
  • the invention is directed to a device as described and claimed herein for use in directly-indicating the presence of a target analyte contained in a bodily secretion present on a subject's skin.
  • FIG. 1 is a cross section of the sweat patch area of the device. This cross section is of an embodiment with only an external membrane (10], a testing pad (20 ⁇ and an active sensor (30 ⁇ ;
  • FIG. 2 is a cross section of the sweat patch area of the device. This cross section is of an embodiment with only an external membrane (10 ⁇ , a testing pad (20 ⁇ , an active sensor (30 ⁇ and a liner (40 ⁇ ;
  • FIG. 3 is a cross section of the sweat patch area of the device. This cross section is of an embodiment with only an external membrane (10 ⁇ , a testing pad (20 ⁇ , an active sensor (30 ⁇ , a liner (40 ⁇ and a second membrane that funnels sweat predominantly towards the active sensor (50 ⁇ ;
  • FIG. 4 is an external view of the sweat patch area of the device. This is an embodiment with only an external membrane (10 ⁇ , a testing pad (20 ⁇ , an active sensor (30 ⁇ and a liner (40 ⁇ ;
  • FIG. 5 is a view of an embodiment of a locking wrist band (100 ⁇ ;
  • FIG. 6 is view of the complete device on a wrist both the dorsal (back of the hand ⁇ and ventral (palm of the hand ⁇ views.
  • An embodiment with room on the wristband for a medical identification nametag is shown (60 ⁇ as a dorsal view.
  • An embodiment with a locking mechanism 70 ⁇ , wristband (80 ⁇ and entire sweat patch sensing area (90 ⁇ is shown as a ventral view.
  • FIG. 7 is a schematic diagram depicting general formulation for imprinted polymers. Polymer was created with the analyte templates, and the templates were later extracted, thus producing binding sites specific to the analytes.
  • FIG. 8 is a diagram showing the isomerization between spiropyran (SP ⁇ and merocyanine (MC ⁇ .
  • SP spiropyran
  • MC merocyanine
  • FIG. 8 shows the isomerization between spiropyran (SP ⁇ and merocyanine (MC ⁇ .
  • SP is colorless and MC is colored.
  • an SP solution shows a yellow color. It is likely that the yellow color is due to the acrylic acid or the existence of some MC isomers.
  • FIG. 9 is an illustration demonstrating a non-imprinted polymer (NIP ⁇ and its non complimentary binding ability for rapid tests and results.
  • FIGS. 10A-10B are photographs showing the set of compared experiments between molecularly imprinted polymer (MIP] and non-imprinted polymer (NIP] systems in the presence of the imprinted molecule of disodium succinate.
  • FIG. 10A shows the experiments of MIP (left] and NIP (center and right] systems in the absence of UV polymerization.
  • FIG. 10B shows the experiments of MIP (left] and NIP (center and right] systems after UV polymerization.
  • FIGS. 11A-11D are photographs showing the color evolution after disodium succinate solution (0.25 M in phosphate buffer] was added into one of the NIP systems (NIP2, right]. The same amount of phosphate buffer was added into the MIP and the other NIP (NIPl, center] the systems. After 50 minutes, the same color as that in the MIP system was developed in the NIP2 system.
  • FIG. 12 is a picture showing cut pieces of solid phase polymers or hydrogels developed from the corresponding solutions shown in FIG. 10A.
  • the MIP polymer or hydrogel (Left] was imprinted with disodium succinate, developed from the corresponding MIP solution (left, FIG. 10A], and incubated in buffer.
  • the NIP polymer or hydrogel (center] was incubated in buffer, developed from the corresponding NIPl solution (center, FIG. 10A].
  • the NIP polymer or hydrogel (right] was incubated in 0.25 M disodium succinate solution, developed from the corresponding NIP2 solution (right, FIG. 10A].
  • FIG. 13 is a graph showing optical absorption spectra of spiropyran solutions with increasing concentrations of disodium succinate molecules.
  • FIG. 14 is a picture showing a comparison of sample color scales of the polymer or hydrogel solutions as the concentration of disodium succinate increases.
  • FIG. 15 is a picture showing the non-imprinted polymer or hydrogel solid forms prepared following UV polymerization of the corresponding solutions. Exposure to UV light does not affect SP molecular photochromic properties, since the samples retain the same color after UV irradiation.
  • FIGS. 16A-16D are a set of pictures showing the color evolution of the solid phase of non-imprinted polymers or hydrogels in the presence of disodium succinate solutions.
  • FIG. 17 is a picture showing the solid phase of non-imprinted polymers or hydrogels incubated in disodium succinate solutions after 24 hours.
  • FIGS. 18A-18B are a set of pictures showing a comparison between the solid phase of non-imprinted polymers or hydrogels after being incubated in disodium succinate for 24 hours and a controlled sample.
  • FIG. 18A is a top view.
  • FIG. 18B is a side view.
  • FIG. 19 is a picture showing the samples of polymer or hydrogel solutions including analyte of disodium succinate (left] and a controlled experiment in the absence of analyte of disodium succinate (right] before UV polymerization.
  • FIG. 20 is a graph illustrating the absorbance spectra of a controlled experiment (green line] and the polymer or hydrogel solutions containing 4 mg of disodium succinate (blue line].
  • the light blue line shows the absorbance spectra of the polymer or hydrogel solution containing 4 mg of disodium succinate after an additional 6 mg disodium succinate were added.
  • FIG. 21 is a picture showing the color changes of the polymer or hydrogel solutions in the absence of disodium succinate by adding different concentrations of acrylic acid.
  • FIG. 22 is a picture showing the color changes of the polymer or hydrogel solutions, acrylic acid concentrations consistent with the solutions shown in FIG. 21, in the presence of disodium succinate (2mg].
  • FIGS. 23A-23B are a set of pictures showing the effect of gold (Au] nanoparticles.
  • FIG. 23A shows polymer or hydrogel solutions of an MIP system (left] and NIP systems (center and right] with gold (Au] nanoparticles.
  • FIG. 23B shows polymer or hydrogel of an MIP system (left] and NIP systems (center and right] containing both SP molecules and gold (Au] nanoparticles initially incubated in buffer (MIP and NIP-center], and disodium succinate solution (NIP-right].
  • FIG. 24 is a picture showing the color change over time of the polymer or hydrogel solutions of MIP system (left] and NIP systems (center and right] after disodium succinate solution was added into one of the NIP systems (NIP2, right].
  • FIGS. 25A-25B are pictures showing a Puritan® flocked swab (FIG. 25A] and a Puritan®'s sponge swab (FIG. 25B].
  • FIGS. 26A-26D are a set of pictures showing a transport system of a
  • FIG. 26A shows a Puritan® sponge swab before colorimetric detection.
  • FIG. 26B shows the Puritan® sponge swab exhibiting a purple color after exposing to disodium succinate and SP solutions successively.
  • FIG. 26C shows the Puritan® sponge swab exhibiting a yellow color after exposing to SP solutions for overnight.
  • FIG. 26D shows the Puritan® sponge swab exhibiting a yellow color after exposing to SP solution for overnight and the remaining SP solution showing no colors.
  • FIGS. 27A-27D are a picture showing that neutralizing buffer is not a suitable transport media for colorimetric detection.
  • FIG. 27A is a neutralizing buffer and SP solution.
  • FIG. 27B is a neutralizing buffer, SP and disodium succinate solution.
  • FIG. 27C is a water, SP and disodium succinate solution.
  • FIG. 27D is an SP and disodium succinate solution.
  • FIGS. 28A-28E are a picture showing that peptone water is a suitable transport media for colorimetric detection.
  • FIG. 28A is a peptone water and SP solution.
  • FIG. 28B is a peptone water, SP and disodium succinate solution.
  • FIG. 28C is a water, SP and disodium succinate solution.
  • FIG. 28D is a SP and disodium succinate solution.
  • FIG. 28E is a peptone water and solid-phase SP.
  • FIGS. 29A-29E are a picture showing that Ames solution is a suitable transport media for colorimetric detection.
  • FIG. 29A is an Ames solution and SP solution.
  • FIG. 29B is an Ames solution, SP and disodium succinate solution.
  • FIG. 29C is a water, SP and disodium succinate solution.
  • FIG. 29D is an SP and disodium succinate solution.
  • FIG. 29E is an Ames solution and solid-phase SP.
  • FIGS. 30A-30E are a picture showing that Butterfield's solution is a suitable transport media for colorimetric detection.
  • FIG. 30A is a Butterfield's solution and SP solution.
  • FIG. 30B is a Butterfield's solution, SP and disodium succinate solution.
  • FIG. 30C is a water, SP and disodium succinate solution.
  • FIG. 30D is an SP and disodium succinate solution.
  • FIG. 30E is a Butterfield's solution and solid-phase SP.
  • FIGS. 31A-31E are a picture showing that Letheen broth is a suitable transport media for colorimetric detection.
  • FIG. 31A is a Letheen broth and SP solution.
  • FIG. 31B is a Letheen broth, SP and disodium succinate solution.
  • FIG. 31C is a water, SP and disodium succinate solution.
  • FIG. 31D is an SP and disodium succinate solution.
  • FIG. 31E is a Letheen broth and solid-phase SP.
  • FIGS. 32A-32E are a picture showing that UTM-RT is a suitable transport media for colorimetric detection.
  • FIG. 32A is a UTM-RT and SP solution.
  • FIG. 32B is a UTM-RT, SP and disodium succinate solution.
  • FIG. 32 C is a water, SP and disodium succinate solution.
  • FIG. 32D is a SP and disodium succinate solution.
  • FIG. 32E is a UTM- RT and solid-phase
  • FIGS. 33A-33I are a set of pictures showing that the present invention of colorimetric detection may be combined with elastic polymers and elastic hydrogels.
  • FIGS. 33A-33C show an elastic hydrogel in the absence of colorimetric detection system showing colorless.
  • FIGS. 33D-33F show an elastic hydrogel having SP showing a yellow color.
  • FIGS. 33G-33I show an elastic hydrogel having SP in the present of disodium succinate showing a pink color.
  • FIG. 34 is a picture showing adenosine triphosphate (ATP] detection in a
  • SP solution Color change was observed upon the addition of an ATP solution to the SP solution.
  • the primary control sample (control] was purple
  • the secondary control sample (w/H20] was dark pink
  • the test sample (w/ATP] shows yellow.
  • the secondary control sample was used to illustrate that the yellow color change observed by ATP interactions was not due to the solvent.
  • the solvent was water (or a PBS buffer] in all the cases.
  • SP solutions were produced by mixing suitable amount of SP and hydroxyethyl methacrylate (HE MA].
  • FIG. 35 is a graph showing UV-Vis spectra for ATP detection according to experiments shown in FIG. 28.
  • the yellow line represents the primary control sample of HEMA+SP at lmg/mL in the absence of water.
  • the pink line represents the secondary control sample of HEMA+SP/DI Water in a 1:1 ratio.
  • the Green line represents the test sample with HEMA+SP/ATP solution in a 1 to 1 ratio. This data corresponds with the proposed MC isomers associated with the "chromic" properties of the SP molecule.
  • FIGS. 36A-36B are a set of pictures and graphs showing the color changes of SP solutions during ATP detection along with peak shifts and peak intensity changes in UV-Vis spectra of the corresponding SP solutions.
  • FIG. 36A shows the color changes of the polymer solutions before (left] and after the addition of an ATP solution (right].
  • FIG. 36B shows the peak shifts and peak intensity changes in UV-Vis spectra of the ATP detection solution (yellow] as compared with those in the control experiment (pink].
  • FIGS. 37A-37B is a set of pictures and graphs showing the color changes of polymer solutions during ATP detection along with peak shifts and peak intensity changes in UV-Vis spectra of the corresponding polymer solutions.
  • FIG. 36A shows the color changes of the polymer solutions before (left] and after the addition of an ATP solution (right].
  • FIG. 36B shows the peak shifts and peak intensity changes in UV-Vis spectra of the ATP detection solution (yellow] as compared
  • FIG. 37A shows the color changes of the polymer solutions before (left] and after the addition of an ATP solution into the polymer solutions (right ⁇ .
  • FIG. 37B shows the peak shifts and peak intensity changes in UV-Vis spectra of the ATP detection solution (test] as compared with those in the control experiment (control ⁇ .
  • a polymer solution formed by mixing suitable amount of acrylamide, ⁇ , ⁇ -Methylene Bisacrylamide, and 4,4-Azobis(4- Cyanovaleric acid ⁇ . Suitable amounts of SP and HEMA were later added into the polymer solution.
  • FIGS. 38A-38B is a set of pictures showing the color changes of solid NIP polymers during ATP detection.
  • the NIP polymer solutions formed by mixing suitable amount of acrylamide, ⁇ , ⁇ -Methylene Bisacrylamide, and 4,4-Azobis(4-Cyanovaleric acid ⁇ . Suitable amounts of SP and HEMA were later added into the NIP polymer solutions.
  • the NIP polymer solutions were polymerized to form solid NIP polymers following UV irradiation.
  • the solid NIP polymer appears to be pink (FIG. 38A, left ⁇ after a PBS solution was added. After the addition of an ATP solution, the solid NIP polymer appears to be yellow (FIG. 38B, right ⁇ .
  • FIGS. 39A-39E are a set of pictures showing time-dependent color changes of solid NIP polymers as shown in FIG. 38.
  • the samples in the left lane are MIP solid polymers
  • the samples in the center lane are NIP solid polymers in the absence of ATP
  • the samples in the right lane are NIP solid polymers in the presence of ATP.
  • the top samples are corresponding polymer solutions before polymerization.
  • the NIP solid polymers or corresponding solutions turn yellow within 1 minute after the addition of ATP solutions.
  • FIG. 40 is a picture showing the color changes of NIP polymer solutions in the presence of different concentration of ATP.
  • the ATP concentrations were 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 mg/mL for the NIP polymer solutions from left to right. Color of the NIP polymer solutions shifts from purple/pink to lighter pink to yellow as the ATP concentrations increase.
  • FIG. 41 is a graph showing UV-Vis spectra of the corresponding NIP polymer solutions as shown in FIG. 34. Peak shift and intensity change can be associated with isomerization of SP.
  • FIG. 42 is a picture showing the color changes of SP solutions in dimethyl sulfoxide (DMSO] after addition of water or ammonium nitrate solutions.
  • SP solutions in DMSO show a blue color (left ⁇ . After addition of DI water into the SP solution in DMSO, the solutions turn pink (center ⁇ . After addition of an ammonium nitrate solution into the SP solution in DMSO, the solution turns yellow (right ⁇ .
  • a SP solution in DMSO was produced by dissolving suitable amount of SP in DMSO.
  • Ammonium nitrate solutions used in the experiments have a concentration of 10 mg/mL in DI water.
  • FIG. 43 is a graph showing UV-Vis spectra of the solutions as shown in
  • FIG. 36 The light blue line represents a primary control sample of SP solution in DMSO at lmg/mL in the absence of water.
  • the green line represents a secondary control sample of SP solution in DMSO and DI water in a 1 to 1 ratio.
  • the dark blue line represents the sample of a SP solution in DMSO and an ammonium nitrate solution in a 1 to 1 ratio. Peak shift and intensity change can be associated with isomerization of SP.
  • FIGS. 44A-44B are a set of pictures and graphs showing ammonium nitrate detection by using SP solutions. After addition of DI water into the SP solution in DMSO, the solutions turn dark pink (left, FIG. 44A ⁇ . After addition of an ammonium nitrate solution into the SP solution in DMSO, the solution turns yellow (right, FIG. 44A ⁇ .
  • FIG. 44B shows UV-Vis spectra of the SP solution in DMSO after addition of DI water (control ⁇ and after addition of an ammonium nitrate solution (test ⁇ .
  • FIGS. 45A-45B are set of pictures and graphs showing ammonium nitrate detection by using polymer and SP solutions.
  • Polymer and SP solutions were produced using similar protocols as discussed above.
  • the polymer and SP solution in the absence of ammonium nitrate shows a pink color (left, FIG. 45A ⁇ .
  • the polymer and SP solution turns yellow (right, FIG. 45A ⁇ .
  • FIG. 45B shows UV-Vis spectra of solutions as shown in FIG. 45A.
  • the pink line represents a control sample of polymer and SP solution.
  • the yellow line represents the sample of polymer and SP solution after addition of an ammonium nitrate solution.
  • FIG. 46 is a set of pictures showing the color changes of solid NIP polymers during ammonium nitrate detection.
  • the samples in the left lane (pink ⁇ are NIP solid polymers in the absence of ammonium nitrate, and the samples in the right lane (yellow] are NIP solid polymers in the presence of ammonium nitrate.
  • Solid NIP polymers were produced following a similar protocol as discussed above.
  • FIGS. 47A-47F are a set of pictures showing time-dependent color changes of solid NIP polymers as shown in FIG. 46.
  • the samples in the left lane are MIP solid polymers
  • the samples in the center lane are NIP solid polymers in the absence of ammonium nitrate
  • the samples in the right lane are NIP solid polymers in the presence of ammonium nitrate.
  • the NIP solid polymers can start turning yellow within 1 minute after the addition of ammonium nitrate solutions.
  • FIG. 48 is a picture showing the color changes of SP solutions in DMSO in the presence of different concentration of ammonium nitrate.
  • the ammonium nitrate concentration changes from 0 to 14 ⁇ L ⁇ .
  • the ammonium nitrate concentration changes from 18 to 500 ⁇ L ⁇ .
  • the solutions in the front row shows a concentration-dependent color change and the color changes from purple/pink to yellow as the concentration of ammonium nitrate increases.
  • FIG. 49 is a graph showing UV-Vis spectra of the solutions as shown in
  • FIG. 42 Peak shift and intensity change can be associated with isomerization of SP. After addition of ammonium nitrate solutions, a new peak was observed at 470 nm.
  • FIG. 50 is a graph showing a Gram positive identification flow chart.
  • analyte refers to a substance, atom or molecule of interest, such as a chemical, which may be detected by a color readout system.
  • color system refers to a solution phase or a solid phase of a Polymer, or a SP solution, which changes color in the presence of an analyte.
  • chromophore refers to a molecule, which undergoes conformation or isomerization change under a condition, leading to a visible color change of the solution having the molecule.
  • photochromism refers to reversible transformation of a chemical species between two forms by the absorption of electromagnetic radiation, where the two forms have different absorption spectra. It may also be described as a reversible change of colour upon exposure to light. As photochromism is just a special case of a photochemical reaction, almost any photochemical reaction type may be used to produce photochromism with appropriate molecular design. Specifically in the present invention, photochromism may be conserved in any suitable forms, such as hydrogel.
  • polymer or hydrogel solution refers to a solution containing at least some of the polymer or hydrogel components, which is soluble in the solution. It may include both MIP and NIP color systems.
  • polymer and “hydrogel”, referring to the same or similar meaning in the invention, are often interchangeable throughout the invention.
  • the terms “polymer” and “hydrogel” include both solid phases of polymers or hydrogels and polymer or hydrogel solutions.
  • colorimetric readout and “colorimetric detection”, referring to the same or similar meaning in the invention, are interchangeable throughout the invention unless indicated otherwise.
  • smart polymers, stimuli-responsive polymers or stimuli- responsive hydrogels are high-performance polymers or hydrogels that change according to the environment they are in. Such materials can be sensitive to a number of factors, such as temperature, humidity, pH, the intensity of light or an electrical or magnetic field and can respond in various ways, like altering color or transparency, becoming conductive or permeable to water or changing shape (shape memory polymers ⁇ . Usually, slight changes in the environment are sufficient to induce greater change in the polymer's properties.
  • Smart polymers and hydrogels may appear in highly specialized applications and everyday products alike. They may be used for the production of hydrogels, biodegradable packaging, and to a great extent in biomedical engineering.
  • One example is a polymer that undergoes conformational change in response to pH change, which may be used in drug delivery.
  • the present invention relates to smart polymers, stimuli-responsive polymers or stimuli-responsive hydrogels for colorimetric detection.
  • kits refers to a device for detection of an analyte in a sample by colorimetric readout.
  • a kit for detection of an analyte in a sample by colorimetric readout may comprise any composition as discussed in the specification, a solid support, and a means for detecting a visible color change of the composition, thereby providing detection of the analyte by colorimetric readout.
  • This disclosure relates to apparatus and methods of analyte detection that involve passive detection, active detection and a security mechanism.
  • This present invention relates to an apparatus and method to use a passive sweat patch sensor with an active detection sensor and security mechanism to show tampering or prevent removal that can detect analytes of interest.
  • the device answers a need for a sweat detection patch that provides an immediate result in addition to functioning as a passive sensor.
  • the device also answers a need for an improved security mechanism that will prevent the removal of the device.
  • One embodiment of such a device would be a wrist band or bracelet that locked onto the human body to prevent removal and contained a sweat patch that contained both active and passive sensing with security tape to prevent tampering with the patch.
  • narcotics There are generally five categories of illicit drugs: narcotics, stimulants, depressants (sedatives], hallucinogens, and cannabis.
  • An illicit drug can be legally produced and prescribed by doctors as well as illegally produced and sold outside of the medical realm.
  • Marijuana pot, Acapulco gold, grass, reefer ⁇ ; tetrahydrocannabinol (THC, marinol ⁇ ; hashish (hash ⁇ ; hashish oil (hash oil ⁇ ; cocaine (coke, snow, crack ⁇ ; chloral hydrate; barbiturates (Amytal, Nembutal, Seconal, phenobarbital ⁇ ; benzodiazepines (Librium, Valium ⁇ ; methaqualone (Quaalude, Mandrax ⁇ ; glutethimide (Doriden ⁇ ; Equanil; Placidyl; Valmid; LSD (acid, microdot ⁇ ; mescaline and peyote; Vicodin; Percocet; amphetamines (Desoxyn, Dexedrine ⁇ ; amphetamine variants (PMA, STP, DOB ⁇ ; phencyclidine (PCP, angel dust, hog ⁇ ; phencyclidine analogues (PCE, PCP
  • Provisional Patent Application 61/513,240 titled NANOCOMPOSITE POLYMERS WITH OPTICAL PROPERTIES, International Patent Application PCT/US2013/065415, filed October 17, 2013, titled SPIROPYRAN-BASED COLORMETRIC DETECTION, U.S. Provisional Application 61/715,994 filed October 19, 2012, and U.S. Provisional Application 61/777,837, filed March 12, 2013.
  • a test could present a color change within the human visual range. It could also produce a change outside the visual range that could be identified for example by UV light or require a black light to be identified. A change outside the visual range would be advantageous since a patient would not be able to notice a change and attempt to remove the device.
  • the disclosure can be calibrated to detect a single analyte of interest or a plurality of them.
  • An example of this could be a single colorimetric assay sensor for cocaine and cocaine metabolites.
  • multiple colorimetric assay sensing areas could also be placed on the device to each detect a different analyte. They could each turn a different color for different analytes.
  • single color could be used and then location of the sensor color change would allow a user to determine which analyte was detected.
  • a space between the patch and the external membrane may also be necessary to prevent interaction of the patch with the membrane material. Patch spacers can be seen in U.S. Patent 6,443,892 as well as Jenkins-Drug Testing in Alternate Biological Specimens.
  • a wide range of materials can be used for the pad that include but are not limited to: Standard medical grade cellulose, cotton, gauze, materials impregnated with salt and water-gel compounds (polymers or hydrogels ⁇ . There are even materials and methods mentioned in Jenkins which can stimulate sweat production.
  • a tamper proof adhesive can be used in the construction of the device.
  • the adhesive can change color when exposed to the air or have capabilities that prevent it from reattaching to the skin once removed.
  • Each sweat collection patch could be individually numbered or have a security code attached to it. This acts to prevent interchanging devices.
  • a single security code feature can be on the patch or a plurality of codes can be on each part of the device.
  • the wristband could include a locking or latching feature that prevents removal.
  • a variety of fasteners or locking devices can be used. They are several simple adhesive or snapping mechanisms for special event bracelets. Medical identification bracelets also have a variety of locking mechanisms that are in use.
  • This device may find use in a hospital setting.
  • the device may be modified to included features similar to a hospital identification bracelet. It can include patient information such as a name or medical information.
  • a bracelet could have the sweat detection portion on the underside or ventral portion of the wrist and medical information on the back-dorsal portion of the wrist to look like a standard medical bracelet. Most medical identification bracelets lock onto a patient's body and must be cut off to be removed.
  • a bracelet could also have RFID capabilities included. Electronic versions of a sensing device could even be built to alter the RFID capabilities or use wireless capabilities to notify a hospital computer that the device had been triggered. Electronic versions of a bracelet could be devised that recognize a colorimetric color change or use an electronic based analyte detection system.
  • This type of analyte detection device may find use in rehabilitation centers or sites of employment.
  • the discrete nature of this device may make it preferable to larger more noticeable sweat patches.
  • a wearer of such a device would likely prefer the discretion that a small watch like bracelet allowed for.
  • An outer ornamental covering could also be added to the sweat patch to completely disguise the nature of the device.
  • This ornamental face could for example be that of a watch.
  • the bracelet could also be modeled on that of a watchband with a discrete locking device.
  • Another example could be a wristband made to look like a piece of jewelry to disguise the device.
  • An active sensing system can be placed directly on the collection pad. It can be positioned so that it is visible when viewed from the outside of the device.
  • other embodiments may include a membrane that funnels sweat to the sensor before the rest of the absorption pad. The positioning of the sensor will also depend on the type of sensor used. A sensor in the design of US patent 7,238,533 that is embedded in nail polish or paint could merely be painted onto the sweat pad. The placement of the sensor is not limited to the embodiments above. Examples of funneling membranes or devices could be a polyurethane membrane or similar material that will not bind small molecules or analytes of interest. The membrane would need to be made non-polar or polar as is necessary.
  • FIG. 1 illustrates a cross section of the sweat patch area of an exemplary device according to the invention.
  • This cross section is of an embodiment with only an external membrane (10], a testing pad (20 ⁇ and an active sensor (30 ⁇ .
  • FIG. 2 is a cross section of the sweat patch area of the device.
  • This cross section is of an embodiment with only an external membrane (10 ⁇ , a testing pad (20 ⁇ , an active sensor (30 ⁇ and a liner (40 ⁇ .
  • FIG. 3 is a cross section of the sweat patch area of the device.
  • This cross section is of an embodiment with only an external membrane (10 ⁇ , a testing pad (20 ⁇ , an active sensor (30 ⁇ , a liner (40 ⁇ and a second membrane that funnels sweat predominantly towards the active sensor (50 ⁇ .
  • FIG. 4 is an external view of the sweat patch area of the device.
  • This is an alternative embodiment with only an external membrane (10], a testing pad (20], an active sensor (30 ⁇ and a liner (40 ⁇ .
  • FIG. 5 is a view of an embodiment of an exemplary locking wrist band (100 ⁇ useful in the present invention.
  • FIG. 6 is view of the complete device of the invention on a wrist both the dorsal (back of the hand ⁇ and ventral (palm of the hand ⁇ views.
  • An embodiment with room on the wristband for a medical identification nametag is shown (60 ⁇ as a dorsal view.
  • An embodiment with a locking mechanism 70 ⁇ , wristband (80 ⁇ and entire sweat patch sensing area 90 ⁇ is shown as a ventral view.
  • the ability to lock this disclosure on to the human body should not be limited to the wrist.
  • a locking device could be used around the forearm or ankle.
  • the present invention provides a device for indicating the presence of a target analyte that is contained in a bodily secretion present on a subject's skin, comprising: (a ⁇ a testing pad to be positioned in contact with a subject's skin; (b ⁇ an external layer to be positioned opposite subject's skin; (c ⁇ a sensor means configured to provide a colorimetric indication of the presence of a target analyte in the subject's bodily secretion while the device is securely-fastened to the subject's skin; and (d ⁇ a securing means in which the testing pad, external layer, and sensor means are contained, said securing means configured to be securely-fastened to the subject's skin.
  • the bodily secretion is preferably the subject's sweat.
  • the target analyte is preferably an illicit drug or metabolite thereof, including, but not limited to, a narcotic, stimulant, depressant, hallucinogen, or cannabis.
  • the device is configured to securely- fasten around the wrist of a human subject.
  • the senor means comprises a molecularly- imprinted polymer material that facilitates the colorimetric indication of the presence of the target analyte in the subject's bodily secretion.
  • the senor means may comprise a nanocomposite material that facilitates the colorimetric indication of the presence of the target analyte in the subject's bodily secretion.
  • the senor means provides a colorimetric indication of the target analyte in the subject's bodily secretion that is in a portion of the electromagnetic spectrum visible to the human eye.
  • the device further includes a secretion impermeable layer positioned between the external layer and the test pad.
  • the device may, alternatively, further include a removable liner which is positioned in contact with the test pad, said removable liner configured for removal before the device is fastened to the subject's skin.
  • the device may further include a secretion wicking layer which is positioned in contact with the sensor means, said secretion wicking layer absorbent to the bodily secretion and in contact with the subject's skin when the device is fastened to the subject's skin.
  • the invention encompasses a method for directly- indicating the presence of a target analyte contained in a bodily secretion present on a subject's skin.
  • a method for directly- indicating the presence of a target analyte contained in a bodily secretion present on a subject's skin includes steps of: (a] securely fastening a device as described and claimed herein on a subject's skin; and (b] examining said device while it is securely-fastened to the subject's skin; wherein said sensor means provides a colorimetric indication of the presence of a target analyte in a bodily secretion present on the skin of the subject.
  • the method is carried out as part of an illicit drug rehabilitation program or illicit drug monitoring program.
  • the method is preferably practiced on a human subject, and the target analyte is an illicit drug or metabolite thereof, including, but not limited to, a narcotic, stimulant, depressant, hallucinogen, cannabis or a metabolite thereof.
  • the device used in the method may comprise a molecularly-imprinted polymer material that facilitates the colorimetric indication of the presence of the target analyte in the subject's bodily secretion.
  • the device may comprise a nanocomposite material that facilitates the colorimetric indication of the presence of the target analyte in the subject's bodily secretion. It is preferred that the device provides a colorimetric indication of the target analyte in the subject's bodily secretion that is in a portion of the electromagnetic spectrum visible to the human eye.
  • the invention is directed to a device as described and claimed herein for use in directly-indicating the presence of a target analyte contained in a bodily secretion present on a subject's skin.
  • This sensor device is not limited to drugs of abuse in humans. Biomarkers can be found in sweat that indicates physiological conditions, genetic defects or disease states. A sensor can be calibrated for these analytes and included in this device. For example, U.S. Patent 6,585,646 discusses the collection of sweat with detectable PSA (Prostate Specific Antigen] levels.
  • PSA Prostate Specific Antigen
  • This device could find use in agricultural applications.
  • the device could be placed onto plants or animals to detect the introduction of analytes of interest. For example, it could detect growth hormones or antibiotics added to live stock. An additional example could be pesticides added to crops.
  • Avoparcin a growth promoter, resulted in the development of vancomycin-resistant enterococci (VRE ⁇ .
  • Enrofloxacin has resulted in the development of ciprofloxacin-resistant strains of Salmonella and Campylobacter.
  • Tylosin contributed to erythromycin-resistant streptococci and staphylococci.
  • Fluoroquinolones have also contributed to the resistant of antibiotics to certain microbes.
  • Growth promoter antibiotics in agriculture can lead to the resistance of bacteria in humans to classic treatment.
  • Drug detection systems can also be applied to race animals, such as horses and dogs, where certain substances may give the animal unfair advantages. There is a great need for the detection of hormones, antibiotics, drugs, and other substances in animals in a multitude of settings.
  • An additional and non-medical application for such an analyte detection device could be for industrial use.
  • the device could be locked onto a pipe to detect leakage.
  • An example of this embodiment could find applications in chemical or food processing plants.
  • molecularly imprinted polymers are polymers that have been processed by a molecular imprinting technique.
  • these polymers possess cavities in the polymer matrix with affinity to a chosen template molecule.
  • a molecular imprinting technique is a laboratory technique commonly adapted by many scientists in the field. Briefly, molecular imprinting is a technique that creates specific recognition sites for a target molecule within a synthetic polymer, and the goal of a molecular imprinting technique is to make an artificial lock for a specific molecule which serves as the key.
  • the formulation generally includes a cross-linking monomer (not shown], and a template (the imprint molecule]. Functional and cross-linking monomers are copolymerized in the presence of a template (the imprint molecule] in a suitable solvent.
  • the template may be the target molecule or any structural derivatives of the target molecule.
  • MIPs show specific binding to the imprint molecules.
  • the functional monomers generally are monomers crossing link with each other.
  • the functional monomers Prior to polymerization, the functional monomers initially form a complex with the template molecules usually by intermolecular interactions such as van der Waals force, electrostatic force, hydrogen bond, ionic bond, etc.
  • the polymer matrix forms around the complex of the functional monomers and the template molecules, so that the monomer-template assembly is held in position by the highly cross-linked three-dimensional rigid structures.
  • cavities are produced within the polymer matrix showing specific sizes, shapes, and chemical functionalities complementary to those of the template molecules. Consequently, the resulting MIPs show specific affinity with the template molecules (the imprint molecules].
  • the resulting MIP contains recognition sites, with shape and functional groups complementary to the imprint molecule.
  • a hydrogel MIP of the present invention requires a specific linker in order to accommodate the geometrical inter molecular distances for which the prior interactive forces may be operative.
  • Hydrogel MIPs of the present invention do not require a specific linker within the imprint to ensure the stability of a MIP specifically for the analyte being used. This is critical as the complication of discovering the specific linker is the rate limiting step in the traditional MIP paradigm. Further, the stabilization of analytes is the restriction factor in non-hydrogel polymers, thus limiting the utility of the traditional methodology of MIPs.
  • a molecular memory is introduced into the resulting MIPs, which is capable of selectively binding specific target molecules.
  • the techniques and the MIPs may be used to fabricate sensors with heightened sensitivity and selectivity.
  • One benefit of the imprinting technique and the MIPs is the capability of controlling the process of uptaking and releasing target molecules simply by varying the experimental conditions such as temperature, pH, or solvents, etc.
  • a variety of monomers may be used in molecular imprinting. For example, different monomers capable of different interactions with imprint molecules may be used. Further, molecular imprinting can be implemented in many systems, such as bulk polymers, beads, membranes, polymer films, polymer sprays and other forms where the proposed device would be useful. In one embodiment, the present invention relates to smart polymers, stimuli-responsive polymers or stimuli-responsive hydrogels for colorimetric detection.
  • the present invention may utilize a polymer or co-polymer comprising one or more polymerizable monomers.
  • a suitable monomer may include acrylamide, 2- ethylphenoxy acrylate, 2-ethylphenoxy methacrylate, 2-ethylthiophenyl acrylate, 2- ethylthiophenyl methacrylate, 2-ethylaminophenyl acrylate, 2-ethylaminophenyl methacrylate, phenyl acrylate, phenyl methacrylate, benzyl acrylate, benzyl methacrylate, 2-phenylethyl acrylate, 2-phenylethyl, methacrylate, 3-phenylpropyl acrylate, 3-phenylpropyl methacrylate, 3-propylphenoxy acrylate, 3-propylphenoxy methacrylate, 4-butylphenoxy acrylate, 4-butylphenoxy methacrylate, 4-butylphenoxy methacryl
  • a suitable monomer may also include benzyl acrylate, phenyl acrylate, naphthyl acrylate, pentabromophenyl acrylate, 2-phenoxyethyl acrylate, 2- phenoxyethyl methacrylate, and 2,3-dibromopropyl acrylate, n-butyl acrylate, n-hexyl acrylate, 2-ethylhexyl acrylate, 2-ethoxyethyl acrylate, 2,3-dibromopropyl acrylate, 1- dihydroperfluorobutyl acrylate, 2-hydroxyethyl acrylate, 2 -hydroxyethyl methacrylate,
  • Imprinting using acrylate-based polymers containing methacrylic or acrylic acid or acrylamide is common in the art. These polymers may form either a noncovalent hydrogen bond or ionic interactions with the imprint molecule.
  • the polymerization may be performed in a dry organic solvent.
  • the suitable organic solvents may include supercritical carbon dioxide, acetone or any other ketones, ethanol and any other alcohols, methyl acetate or any other acetates, tetrachloroethylene, toluene, turpentine, hexane, petrol ether, or any other alkanes, 1,4- Dioxane or any its derivatives, cyclopentane or any other cycloalkanes, Benzene or any of its derivatives, chloroform, dichloromethane, diethyl ether or any other ethers, tetrahydrofuran, dimethylformamide, acetonitrile, dimethyl sulfoxide, propylene carbonate, formic acid or any other organic acid, and any other organic compounds.
  • the polymerization may also be performed in water or another inorganic solvent.
  • the suitable inorganic solvents may include a solvent other than water that is not an organic compound. Common examples may include liquid ammonia, liquid sulfur dioxide, sulfuryl chloride and sulfuryl chloride fluoride, phosphoryl chloride, dinitrogen tetroxide, antimony trichloride, bromine pentafluoride, hydrogen fluoride, pure sulfuric acid and any other inorganic acids.
  • Polymers may be prepared with impressive selectivity towards low molecular weight or stereochemistry.
  • the molecules may include, but not limited to, small or low molecular weight organic molecules, inorganic ions, biologic pathway molecules, industrial solvents, and explosive or environmentally toxic chemicals.
  • Polymers may be prepared with impressive selectivity towards small molecules including physiologically active drugs.
  • Pharmacological structures imprinted for include classes of drugs (imprint antigens], such as Theophylline, Diazepam, Morphine, S-Propranolol and Cortisol. Many other molecules can be imprinted and thus, this application is not limited to the molecules named here.
  • the indicator mechanism of the present invention may be operated via any suitable process resulting in a permanent and detectable change in the template or in the indicator (sensor] mechanism.
  • a subsequent polymerization in the presence of a cross-linker, a cross-linking reaction or other process results in the formation on an insoluble matrix in which the template sites reside.
  • the permanent and detectable change may be the color change of the polymers matrix solution and the resulting polymers, which is caused by the shift of UV- Vis absorption bands of the indicator compounds under certain conditions.
  • Hydrogels and Molecularly Imprinted Hydrogels are insoluble networks of cross-linked polymer structures comprising hydrophilic homo- or hetero- copolymers. Hydrogels are highly absorbent natural or synthetic polymers and they can contain over 99.9% water. Hydrogels may also include macroporous structures. Hydrogel MIPs of the present invention do not require a specific linker within the imprint to ensure the stability of a MIP specifically for the analyte being used. This is critical as the complication of discovering the specific linker is the rate limiting step in the traditional MIP paradigm. Further, the stabilization of analytes is the restriction factor in non-hydrogel polymers, thus limiting the utility of the traditional methodology of MIPs.
  • molecularly imprinted polymers may be molecularly imprinted hydrogels.
  • Molecularly imprinted polymers (MIPs] or hydrogels may be developed from a polymer or hydrogel solution following the additional procedures.
  • the additional procedures may include UV light irradiation, temperature change, solvent change, evaporation of solvents, pH change, addition of chemicals, change of salt concentration, etc.
  • the present invention is about a solution phase of the polymer or hydrogel, including the polymer or hydrogel-forming solution, which changes colors upon addition of a chemical or an analyte.
  • the present invention is about solid phases of the polymer or hydrogel, which changes colors upon addition of a chemical or an analyte.
  • the solid phases of polymers or hydrogels may be films or any other solid form.
  • the present invention is about a SP solution, which changes colors upon addition of a chemical or an analyte.
  • Photochromism of Spiropyrans and Merocyanines Isomerization researchers have recently examined the ability of a chromophore to adhere a cation, and release it upon UV exposure. Photochromism and solvatochromism are the two key fields on which the intense researches have been focused. Photochromism is the reversible transformation of a chemical species between two forms by the absorption of electromagnetic irradiation, where the two forms have different absorption spectra. Photochromism usually refers to compounds that undergo a photochemical reaction where an absorption band in visible region of the electromagnetic spectrum changes dramatically in strength or wavelength. Solvatochromism refers to the ability of a chemical substance to change color due to a change in solvent polarity.
  • FIG. 8 a generic diagram of photochromism of spiropyran and merocyanine isomerization is depicted.
  • spiropyran SP, left
  • the isomerization takes place where the bond between the oxygen of the benzopyran and the spiro-carbon breaks, leading to the opening of the pyran ring.
  • the spiro-carbon has an sp2 hybridization, showing a planar structure, and the rotation of aromatic group leads to the alignment of ir-orbitals of the spiro-carbon with the rest of the molecule.
  • the resulting form of merocyanine (MC, right], a zwitterion form, possesses a highly conjugated double-bond system, with a capability of absorbing photons in the visible light region of the electromagnetic spectrum. Therefore, merocyanine (MC] appears to be colored.
  • photochromism of spiropyran and merocyanine isomerization may be used as the indicator mechanism of color readout. It is found that the interaction of certain monomers and molecules with the two forms of SP or MC may drive the equilibrium towards the colorless or colored states, forming the fundamentals of optical readout system in the present invention. It is believed by some researchers that different MC isomers yield different colors. Thus, photochromism of SP and MC isomerization may enable a color readout system showing different colors.
  • the invention is a non-imprinted polymer (NIP] capable of color readout.
  • FIG. 9 shows a simple illustration of a non-imprinted polymer (NIP ⁇ .
  • NIPs do not possess specific binding ability to a predesigned analyte or chemical.
  • NIPs may be highly desired under a simple environment where specificity is not required and a fast result is highly expected. These types of environments may be seen mostly in production settings where the molecule to be detected is very specific and delays in this detection could be very costly, thus making the rapid detection a top priority.
  • Field testing would be another beneficial application of the product as some companies require validation testing before importation. While the non-imprinted polymer lacks specificity, it does still offer a certain level of retention.
  • an MIP color system including a polymer or hydrogel forming solution, disodium succinate as the analyte, and spiropyran as the chromophore, may be created.
  • NIPs non- imprinted Polymer color systems
  • SP chromophore Spiropyran
  • the use of disodium succinate may be served as an analog to the Gamma Hydroxy Butyrate metabolite, succinic acid. The results are illustrated in FIGS. 10A-10B.
  • the MIP color system of the polymer or hydrogel solution in the presence of disodium succinate exhibits a distinguishably red color as compared with the other two NIP color systems (NIP1 and NIP2], showing a yellow- orange color.
  • UV light does not affect SP molecular photochromic properties. It is desired for a color readout system that a color change occurs only through the interaction of the analyte with the SP molecule, and other environmental parameters such as UV light or sunlight, may not affect the interaction.
  • FIG. 10B following UV irradiation, the colors of the MIP color system and the NIP color systems remained unchanged, confirming that exposure to UV light does not affect SP molecule photochromic properties after polymerization.
  • the solid forms of polymers or hydrogels may be produced for color readout systems.
  • FIG. 12 the cutting pieces of polymers or hydrogels formed from the corresponding polymer or hydrogel solid forms in buffer and disodium succinate solutions described in FIG. 5D are depicted.
  • polymers or hydrogels produced from the corresponding MIP color system and the NIP2 color system with later addition of disodium succinate show the same red color as those of the solutions.
  • the NIPl color system in the absence of disodium succinate produced the polymer or hydrogel showing the original yellow- orange color.
  • FIG. 13 shows optical absorption spectra of SP solutions comprising phosphate buffer solution (PBS], acrylic acid, SP with different concentrations of disodium succinate measured by an UV-Vis spectrometer. While FIG.
  • PBS phosphate buffer solution
  • FIG. 13 shows optical absorption spectra of SP solutions comprising phosphate buffer solution (PBS], acrylic acid, SP with different concentrations of disodium succinate measured by an UV-Vis spectrometer. While FIG.
  • the UV-Vis spectra of the SP solutions demonstrate that an absorption band centered at 370 nm, due to SP, shows a decreasing intensity, and the other absorption band centered at 530 nm, due to MC, has an increasing intensity.
  • the UV-Vis spectra were further visualized by sample color scales of the polymer and hydrogel solutions having SP and different concentrations of disodium succinate shown in FIG. 14. The concentrations of disodium succinate increase from left to right.
  • UV-Vis spectra and sample color scales further confirm that the interaction between the analyte (disodium succinate] and the chromophore (SP and MC] is indeed the cause of the color change in the SP solutions.
  • the increasing intensity of the absorption band at 530 nm corresponds to the increased color intensity of the solutions.
  • a NIP color system including a polymer or hydrogel solution in the absence of analyte (disodium succinate], and with spiropyran as the chromophore, may be created. Similar to the MIP color system, a NIP color system demonstrates novel utilities in detecting an analyte, such as disodium succinate.
  • the NIP color systems of the polymer or hydrogel solutions including the chromophore (SP and MC] were made following a similar procedure to the above experiments. The initially prepared polymer or hydrogel solutions showed a yellow- orange color.
  • a NIP color system including a polymer or hydrogel solution in the absence of analyte (disodium succinate], and with spiropyran as the chromophore, may also be used to detect an analyte.
  • a solid phase of a NIP color system having a polymer or hydrogel and a chromophore (SP and MC] in the absence of analyte (disodium succinate] may be produced.
  • the polymer or hydrogel in a solid phase may demonstrate the same characteristic behavior of color change in the presence of an analyte.
  • the solid phase of a NIP color system may be produced from the corresponding polymer or hydrogel solutions as shown in FIG. 15 by a suitable means.
  • the suitable means may include solvent evaporation, heating, or any other methods known to those skilled in the art.
  • the polymer or hydrogel of a NIP color system in a solid phase was made, and three of the samples were each put into a solution of disodium succinate (the first three samples from left], and another sample was put into a phosphate buffer solution as a controlled experiment (the right ⁇ .
  • the polymer or hydrogel in a solid phase in the solution of disodium succinate demonstrates a clear color change from yellow-orange to red.
  • the polymer or hydrogel in a solid phase remained the yellow-orange color.
  • FIGS. 18A-18B show the samples of the polymer or hydrogel in a solid phase after taken out from the solutions. As shown in FIGS.
  • the samples of the polymer or hydrogel in a solid phase which were in the solution of disodium succinate show the same red color
  • the sample of the polymer or hydrogel in a solid phase which was in phosphate buffer (the right] remained the yellow-orange color. Since the color changes are due to the interaction between the analyte (disodium succinate] and the chromophore (SP and MC], showing the penetration of analyte into the solid phase of the polymer or hydrogel, these observations demonstrate that the polymer or hydrogel in a solid phase may also be used as a color readout system to detect an analyte.
  • FIG. 20 shows the UV-Vis spectra observed on the polymer or hydrogel solution of the NIP system and on the sample of a controlled experiment.
  • an absorption band centered at 530 nm appears, which was due to MC in the presence of disodium succinate (4 mg in lOmL; blue line].
  • disodium succinate 4 mg in lOmL; blue line.
  • the intensity of the absorption band at 530 nm increases (lighter blue line], demonstrating an increasing interaction between the analyte of disodium succinate and the chromophore (SP and MC].
  • FIG. 21 shows a color scale of the polymer or hydrogel solutions of a NIP system including the chromophore (SP and MC] in the absence of the analyte of disodium succinate with a different concentration of acrylic acid. As shown in FIG. 21, in the absence of acrylic acid, the polymer or hydrogel solution appears to be red (the left].
  • a NIP color system may be built by including a polymer or hydrogel solution, a chromophore (SP and MC], and a different concentration of acrylic acid in the absence of the analyte of disodium succinate.
  • acrylic acid derivatives may substitute acrylic acid in NIP color systems.
  • the acrylic acid derivatives may include acrylic acid esters, such as ethyl acrylate, butyl acrylate, 2-ethylhexyl acrylate, sorbyl acrylate, 2-( dimethylamino]- ethyl acrylate, 3,3-dimethoxypropyl acrylate, 2,2,3,3,4,4,4-heptafluorobutyl acrylate, 2- cyanoethyl acrylate, 4-tluorophenyl acrylate, chloroethyl acrylate, 2-(propen-l-yloxy]- ethyl acrylate, phenyl acrylate, allyl acrylate, hydroxyethylmethacrylate (HEMA], acrylamides, such as ⁇ , ⁇ -dimethylacrylamide, and acrylonitrile.
  • acrylic acid esters such as ethyl acrylate, butyl acrylate, 2-ethylhexyl acrylate, sorby
  • FIGS. 23A-23B show the color changes of both MIP (left] and NIP systems (center and right] in the presence of Au nanoparticles.
  • FIG. 23A the polymer or hydrogel solutions of an MIP system (left] and NIP systems (center and right] in the absence of chromophore (SP and MC] were made.
  • FIG. 23B shows the color changes of both MIP (left] and NIP systems (center and right] in the presence of Au nanoparticles and the chromophore (SP and MC]. As shown in FIG.
  • the polymer or hydrogel solid forms in solutions for the MIP system illustrates a red color (left], while the solutions for the NIP systems appear to be brown (center and right ⁇ .
  • Au nanoparticles have an effect the NIP systems by changing the color from yellow-orange to brown.
  • the polymers here are representative of the initial time period of solution with buffer and disodium succinate solutions being added respectively.
  • FIG. 24 shows an additional experiment following those in FIG. 23B.
  • the analyte of disodium succinate was added into one of the NIP systems (right] while the other NIP system (center] and the MIP system (left] remained the same as those in FIG. 23B.
  • the polymer or hydrogel solid forms in solutions (right] changes to a same red color as that in the MIP system (left] after 50 minutes.
  • a molecularly imprinted polymer (MIP] system may be suited for an environment requiring specificity, and a non-imprinted polymer (NIP] system, possessing sufficient analyte retention levels, may offer prescreening and early detection capabilities. Both systems may offer a visible readout, which is yet to be accomplished by others in the art.
  • a NIP system may serve well as a prescreening device or a technology to be utilized under an analyte-specific environment. For example, if the goal were to detect bacterial metabolites in a food production setting, as a prescreening device, the NIP system would yield a color based on the overall concentration of metabolites. If no color change were detected, no further testing would be required.
  • MIP polymer
  • the present invention of the color readout systems of MIPs and NIPs may be very well suited for detection of metabolites the field of metabolomics, whereas other technologies were more geared towards proteomics and genomics.
  • the present invention of the color readout systems of MIPs and NIPs may be very well suited for many sectors in industry that desire faster detections in the field of detection. For example, USDA consistently researches improvements in health and safety in the food industry, and the detection represents a large part of advancement in the field. Future areas of interest for the invention may include the fields of E. coli, toxins (water safety], and food packaging safety.
  • the present invention relates to a SP solution, which is capable of undergoing a color change when the solution is contacted with an analyte, thereby providing detection of the analyte by colorimetric readout.
  • the SP solution comprises phosphate buffer solution (PBS], acrylic acid, SP, and disodium succinate as the analyte.
  • PBS phosphate buffer solution
  • the SP solution may be made in any suitable solvent.
  • the suitable solvents may include water, DMSO, acetone, alcohol, DMF, pyridine, or any other solvents capable of dissolving SP, acrylic acid, and disodium succinate.
  • the solvent is water.
  • SP is initially dissolved in acrylic acid, and the resulting solution is then diluted with PBS.
  • the present invention relates to a transport system, on which a colorimetric detection may be visualized.
  • transport system refers to a system which can carry at least a certain amount of the substrates after it contacts a solution having substrates.
  • substrates we mean chemical compounds in a solution.
  • the substrates may include SP, disodium succinate, phosphate buffer, acrylic acid, MIPs, NIPs, nanoparticles, and others.
  • a transport system may generally have sponge-like or any other macroporous structures, providing a means to hold substrates.
  • the transport system may be flocked or sponge swabs, and more preferably, the transport system may be flocked or sponge swabs having white or other light colors.
  • the transport system may be Puritan® flocked or sponge swabs.
  • the substrates may be transported by inserting the transport system into a solution of disodium succinate and an SP solution successively.
  • the transport system may be first inserted into the solution of disodium succinate, and it may be then inserted into the SP solution.
  • the successive insertion into the solution of disodium succinate and the SP solution may be performed once.
  • the successive insertion into the solution of disodium succinate and the SP solution may be performed twice or multiple times.
  • the present invention relates to transport medias as suitable platforms for colorimetric detection.
  • transport media refers to a media selected from the group consisting of temporary storage of specimens being transported to the laboratory for cultivation, those maintaining the viability of all organisms in the specimen without altering their concentration, those containing only buffers and salt, those lack of carbon, nitrogen, and organic growth factors so as to prevent microbial multiplication, and those medias used in the isolation of anaerobes free of molecular oxygen.
  • transport media may include, but not limited to, thioglycolate broth for strict anaerobes, Stuart Transport medium (a non-nutrient soft agar gel containing a reducing agent to prevent oxidation, and charcoal to neutralize], certain bacterial inhibitors (for gonococci, and buffered glycerol saline for enteric bacilli], or Venkat-Ramakrishnan (VR] medium for V. cholerae.
  • thioglycolate broth for strict anaerobes Stuart Transport medium (a non-nutrient soft agar gel containing a reducing agent to prevent oxidation, and charcoal to neutralize]
  • certain bacterial inhibitors for gonococci, and buffered glycerol saline for enteric bacilli
  • VR Venkat-Ramakrishnan
  • transport medias as suitable platforms for colorimetric detection.
  • the present invention may be applied to any transport medias of non-neutralizing buffer.
  • a suitable transport media may be chosen from, but not limited to, the group consisting of peptone water, Ames solution, Butterfield's solution, Letheen Broth, and UTM-RT.
  • such incorporation of the colorimetric detection into transport medias may provide a broad color spectrum for a readout scale.
  • the present invention relates to elastic polymers and elastic hydrogels for colorimetric detection.
  • the present invention of colorimetric detection may be combined with elastic polymers and elastic hydrogels.
  • Elastic polymers and elastic hydrogels sometimes referred to as elastomers, are materials with viscoelasticity (colloquially "elasticity" ⁇ , generally having low Young's modulus (E] and high yield strain compared with other materials.
  • Elastic polymers may be amorphous polymers existing above their glass transition temperature, so that considerable segmental motion is possible. At ambient temperatures, elastic polymer may thus be relatively soft (E ⁇ 3MPa] and deformable but they recover their shape readily when the applied external stress is removed.
  • Their primary uses may be for, but not limited to, seals, adhesives and molded flexible parts. In the proposed embodiment, they could add sensing capabilities for detection of molecules.
  • Colorimetric detection of the present invention may be suitable for any elastic polymers and elastic hydrogels.
  • the Examples describe elastic polymers and elastic hydrogels for colorimetric detection.
  • An elastic polymer and elastic hydrogel for colorimetric detection may provide additional functions of colorimetric readout while the elastic properties are maintained.
  • an elastic polymer and elastic hydrogel for colorimetric detection may be made by combining all the chemicals of colorimetric detection and those of elastic polymers and elastic hydrogels before polymerization.
  • an elastic polymer and elastic hydrogel for colorimetric detection may be made by adding all the chemicals of colorimetric detection into the elastic polymer and elastic hydrogel.
  • the present invention relates to a kit for detection of an analyte in a sample by colorimetric readout.
  • the kit may comprise any chemical compositions for colorimetric detection as discussed above.
  • the kit may comprise a solid support.
  • a solid support may comprise any suitable means of support.
  • the solid support may be a transport system as discussed above.
  • the kit may further comprise a transport media.
  • a transport media may comprise any suitable medias.
  • the transport media may be any media as discussed above.
  • the kit may further comprise an elastic polymer.
  • An elastic polymer may comprise any suitable polymers.
  • the elastic polymer may be any polymer as discussed above.
  • the present invention relates to systems of a SP solution consisting of SP and a solvent and methods of using such system for colorimetric detection.
  • Any suitable solvent may be used for the SP solution.
  • a suitable solvent may be selected from the group consisting of water, DMSO, acetone, alcohol, DMF, hydroxyethyl methacrylate (HEMA ⁇ , and pyridine.
  • a suitable solvent may be selected from the group consisting of DMSO, acetone, alcohol and hydroxyethyl methacrylate (HEMA ⁇ .
  • a suitable solvent for the present invention may be a mixture combination of two or more solvents. The two or more solvents may be mixed initially before they are used to dissolve SP.
  • the two or more solvents may also be used in a subsequent manner, in which one or more of the solvents are used to dissolve SP to make a SP solution, and other solvents are subsequently added into the solution.
  • the present methods of colorimetric detection using a SP solution consisting of SP and a solvent may show different color changes depending on the manners in which two or more solvents are added, e.g., simultaneously, subsequently, and the sequence of the solvents.
  • SP solution consisting of SP and a solvent may be applicable to any of the above methods, devices, and kits.
  • the present SP solution consisting of SP and a solvent may be used in a kit or a transport system as discussed above.
  • the present SP solution consisting of SP and a solvent may be used to detect a bio-molecule such as a nucleoside triphosphate.
  • a bio-molecule such as a nucleoside triphosphate.
  • the nucleoside triphosphate to be detected may be Adenosine-5'-triphosphate (ATP ⁇ .
  • FIGS. 34, 35, and 36A-36B and the Examples show colorimetric detection of ATP by using a SP solution consisting of SP and a solvent.
  • the present SP solution consisting of SP and a solvent may further comprise polymers for colorimetric detection of a bio-molecule such as ATP. Any polymers may be applicable for the present invention.
  • the present SP solution may be applicable to a solution-phase detection.
  • FIGS. 37A-37B and the Example show a solution-phase colorimetric detection of ATP by using a SP solution comprising SP, a solvent and polymers.
  • the present SP solution may be applicable to a solid-phase colorimetric detection.
  • FIGS. 38A-38B and 39A-39B and the Examples show a solid-phase colorimetric detection of ATP by using a SP solution comprising SP, a solvent and polymers.
  • the present colorimetric detection by using a SP solution may be sensitive to the concentration of the analyte. For example, as shown in FIGS. 40-41, visually color changes may be detectable in the presence of 2 mg/mL ATP.
  • the present SP solution and methods of using such solution may be applicable to detect explosives.
  • the present invention may be suitable for detecting any substances related to explosives.
  • the chemical compound ammonium nitrate, the nitrate of ammonia with the chemical formula NH4N03 is a white crystalline solid at room temperature and standard pressure. It is commonly used in agriculture as a high-nitrogen fertilizer, and it has also been used as an oxidizing agent in explosives, including improvised explosive devices. It is the main component of ANFO, a popular explosive, which accounts for 80% explosives used in North America. Ammonium-nitrate-based explosives were used in the Oklahoma City bombing and in the 2011 Delhi bombing, 2013 India blasts and the bombing in Oslo 2011. A simple method for detecting such chemical compounds would be of great value for anti-terrorism.
  • FIGS. 40-43, 44A-44B and 45A-45B and the Examples show solution- phase colorimetric detection of ammonium nitrate by using a SP solution.
  • the present colorimetric detection of ammonium nitrate may be also applicable to solid-phase detections.
  • FIGS. 46 and 47A-47F and the Examples show solid-phase colorimetric detection of ammonium nitrate by using a SP solution.
  • the present colorimetric detection of ammonium nitrate by using a SP solution may be sensitive to the concentration of ammonium nitrate. For example, as shown in FIGS. 48 and 49, visual color changes may be detectable in the presence of tens ⁇ , ammonium nitrate.
  • Applicants envision that the present colorimetric detection may be applicable to detect bacteria.
  • the above methods for detecting ATPs may be used to detect bacteria which contain ATPs.
  • Gram staining is a method of differentiating bacterial species into two large groups (gram-positive and gram- negative ⁇ . Gram staining differentiates bacteria by the chemical and physical properties of their cell walls by detecting peptidoglycan, which is present in a thick layer in gram- positive bacteria.
  • dyes are generally used. A gram-positive species may result in a purple-blue color while a gram-negative species may result in a pink-red color.
  • the present SP solution may provide an easy way to differentiate Gram negative bacteria and positive bacteria by colorimetric detection.
  • a Gram negative bacterium may exhibit different color from that of a Gram positive bacterium.
  • Gram negative bacteria may comprise Salmonella sp., Campylobacter jejuni, Yersinia enterocolitica, Shigella sp., Vibrio parahaemolyticus, Coxiella burnetii, Mycobacterium bovis, Brucella sp., Vibrio cholerae serogroup 01 and 0139, Vibrio cholerae serogroup non-01 and non-0139, Vibrio vulnificus, Cronobacter (Enterobacter sakazakii ⁇ sp., Aeromonas hyhrophila and other sp., Plesiomonas shigelloides, Miscellaneous bacterial enterics, Francisella tularensis, Enterotoxigenic Escherichia coli (ETEC ⁇
  • Gram positive bacteria may comprise Clostridium perfringens, Clostridium botulinum, Staphylococcus aureus, Bacillus cereus and other Bacillus sp., Listeria monocytogenes, Streptococcus sp., and Enterococcus.
  • Molecularly Imprinted Hydrogels and Detections via Spiropyran Molecules were produced in 20 mL glass vials with the compositions of 5M acrylamide, 0.5 M acrylic acid, 0.1 M N,N'-methylene- bisacrylamide, 0.25 M disodium succinate, and 0.0178 g 4,4'-azobis(4-cyanovaleric acid ⁇ .
  • the copolymerization reactions of acrylamide and bisacrylamide were initiated by 4,4'-azobis(4-cyanovaleric acid ⁇ .
  • the high ratio of acrylamide to bisacrylamide (20/1 ⁇ leads to the formation of the polymer networks where bisacrylamide serves as the bridge junctions.
  • the MIP color system of the polymer or hydrogel solutions and the polymer or hydrogel solid forms in solutions in the presence of disodium succinate exhibits a distinguishably red color as compared with the other two NIP color systems (NIP1 and NIP2 ⁇ , showing a yellow- orange color.
  • UV light does not affect SP molecular photochromic properties. It is desired for a color readout system that a color change occurs only through the interaction of the analyte with the SP molecule, and other environmental parameters such as UV light or sunlight, may not affect the interaction.
  • a colorimetric system was initially include in the MIP polymer system. Following UV irradiation, the colors of the MIP color system and the NIP color systems remained unchanged, confirming that exposure to UV light does not affect SP molecule photochromic properties after polymerization.
  • Non-Imprinted Hydrogels and Detections via Spiropyran Molecules Non- imprinted polymers (NIPs ⁇ were made in 20 mL glass vials with the compositions of 5M acrylamide, 0.5 M acrylic acid, 0.1 M ⁇ , ⁇ '-methylene-bisacrylamide, and 0.0355g 4,4'- azobis(4-cyanovaleric acid ⁇ . The copolymerization reactions of acrylamide and bisacrylamide were initiated by 4,4'-azobis(4-cyanovaleric acid ⁇ . The high ratio of acrylamide to bisacrylamide (20/1 ⁇ leads to the formation of the polymer networks where bisacrylamide serves as the bridge junctions.
  • the initially prepared polymer or hydrogel solutions showed a yellow-orange color.
  • ten samples having the same concentrations of the polymer or hydrogel and the chromophore were irradiated by UV light, and the resulting polymer or hydrogel solid forms in solutions were depicted in FIG. 15.
  • the polymer or hydrogel solid forms in solutions remained consistent in color (yellow- orange], illustrating no additional attribution to the photochromic properties of the SP- MC system.
  • a NIP system using Au nanoparticles instead of SP was made in 20 mL glass vials with the compositions of 5M acrylamide, 0.5 M acrylic acid, 0.1 M ⁇ , ⁇ '- methylene-bisacrylamide, and 0.0178 g 4,4'-azobis(4-cyanovaleric acid].
  • the above compounds were dissolved in a phosphate buffer with a pH of 7.4 with a total volume of 5 mL.
  • Corresponding MIP and NIP systems were also made in the presence of SP, and similar procedures were used by later addition of 1.0 mg SP molecule to each vial and the solutions were sonicated for at least 5 minutes.
  • FIG. 23A-23B shows the color changes of both MIP (left] and NIP systems (center and right] in the presence of Au nanoparticles.
  • the polymer or hydrogel solutions of an MIP system (left] and NIP systems (center and right] in the absence of chromophore (SP and MC] were made. Even in the absence of a chromophore, the interaction between the analyte of disodium succinate and Au nanoparticles led to a red color of the resulting solution for the MIP system (left].
  • FIG. 23B shows the color changes of both MIP (left] and NIP systems (center and right] in the presence of Au nanoparticles and the chromophore (SP and MC].
  • the polymer or hydrogel solid forms in solutions for the MIP system illustrates a red color (left], while the solutions for the NIP systems appear to be brown (center and right ⁇ .
  • Au nanoparticles have an effect the NIP systems by changing the color from yellow-orange to brown.
  • FIG. 24 shows an additional experiment following those in FIG. 23B.
  • the analyte of disodium succinate was added into one of the NIP systems (right] while the other NIP system (center] and the MIP system (left] remained the same as those in FIG. 23B.
  • the polymer or hydrogel solid forms in solutions (right] changes to a same red color as that in the MIP system (left] after 50 minutes.
  • Puritan® Flock or Sponge Swabs As Transport Systems for Colorimetric Detection. Both Puritan® flocked or sponge swabs may be used as transport systems for colorimetric detection. Both a disodium succinate solution and a SP solution were made according to the methods discussed above. Specifically, a disodium succinate (DS] Solution was made after a suitable amount of disodium succinate dissolved in phosphate buffer solution or dewater with a concentration depending on the experiment.
  • a polymer solution refers to the mixture of polymer components (buffer, monomers, crosslinker, initiator, analyte when applicable, SP when applicable] before polymerization.
  • a polymer/solid polymer refers to the mixture of polymer components (buffer, monomers, crosslinker, initiator, analyte when applicable, SP when applicable] AFTER polymerization.
  • a SP solution refers to a mixture consisting of only lmg SP dissolved in 685uL acrylic acid after the addition of 20mL of PBS. A 20mL SP solution was thus produced.
  • both flocked and sponge swabs were inserted into the disodium succinate solution and the SP solution successively.
  • the successive insertion into disodium succinate and SP solutions were repeated three times, and the flocked swab was rinsed with water each time between each trial.
  • both flocked and sponge swabs showed a white color initially.
  • the SP solution remains its original color of dark yellow, while the disodium succinate solution remain colorless.
  • the flocked swab's color changed from white to faint yellow. This process was repeated three times. The only difference in the three trials was the time period the swab was left in each solution. During the first trial it was less than 5 seconds. During the 2nd, it was about 10 seconds and during the 3rd, it was about 20 seconds.
  • the flocked swab After the third insertion into the disodium succinate solution, the flocked swab turned red-pink (not shown], indicating that a colorimetric detection is accomplished and visualized on the flocked swab. Upon a further insertion into the SP solution, the red-pink flocked swab turned back to yellow.
  • FIGS. 26A-26D a sponge swab appeared to be white initially (FIG. 26A ⁇ . After a first insertion into disodium succinate and a second insertion into SP solutions, the sponge swab turned purple (FIG. 26b] and the SP solution remained yellow (not shown ⁇ .
  • Transport Medias as Suitable Platforms for Colorimetric Detection Various transport medias was tested for suitable platforms for colorimetric detection. It appeared that most of the transport medias except for the neutralizing buffer are suitable for colorimetric detection in the present invention.
  • a neutralizing buffer comprises monopotassium phosphate (0.0425g/L], sodium thiosulfate (0.16g/L], and aryl sulfonate complex (5.0g/L ⁇ .
  • a solution comprising neutralizing buffer and SP solutions showed a yellow color (FIG. 21A ⁇ .
  • a solution comprising neutralizing buffer, SP and disodium succinate solutions showed a yellow color (FIG. 27B ⁇ .
  • solutions comprising SP and disodium succinate solutions with (FIG. 27C] or without (FIG. 27D] additional water show red-pink colors.
  • a neutralizing buffer is not a suitable platform for colorimetric detection.
  • Peptone water was tested for colorimetric detection.
  • Peptone water may include any kind of various water-soluble protein derivatives obtained by partial hydrolysis of a protein by an acid or enzyme during digestion and used in culture media in bacteriology.
  • Peptones may be derived from animal milk or meat digested by proteolytic digestion.
  • Peptone water may further include fats, metals, salts, vitamins and many other biological compounds.
  • Peptone water may be used in nutrient media for growing bacteria and fungi.
  • a buffered Peptone water comprises Peptone (10.0 g/L], sodium chloride (5.0 g/L], disodium phosphate (3.5 g/L], monopotassium phosphate (1.5 g/L], at final pH of 7.2 ⁇ 0.2 at 25°C.
  • a solution comprising peptone water and SP solutions showed a yellow color (FIG. 28A ⁇ .
  • a solution comprising peptone water SP and disodium succinate solutions showed a red-pink color (FIG. 28B ⁇ .
  • FIG. 28E an experiment of directly mixing of a solid phase of SP and peptone water showed that the solid phase of SP is not very soluble in the peptone water and the solution remained yellow.
  • Ames solution was tested for colorimetric detection.
  • Ames solution comprises inorganic salts of calcium chloride (0.1275 g/L], magnesium sulfate (0.1488 g/L], potassium chloride (0.231 g/L], potassium phosphate monobasic (anhydrous] (0.068 g/L], sodium chloride (7.01 g/L], amino acids of L- Alanine (0.0024 g/L], L-Arginine ⁇ HC1 (0.00421 g/L], L-Asparagine (anhydrous] (0.00084 g/L], L-Aspartic Acid (0.00012 g/L], L-Cystine ⁇ 2HC1 (0.000065 g/L], L- Glutamine (0.073 g/L], L-Glutamic Acid (sodium] (0.001183 g/L], Glycine (0.00045 g/L], L-Histidine ⁇ HC1 ⁇ H20
  • a solution comprising Ames and SP solutions showed a yellow color (FIG. 29A]. Contrastingly, a solution comprising Ames, SP and disodium succinate solutions showed a red-pink color (FIG. 29B]. This observation, consistent with those standard colorimetric detections (FIG. 29C-29D], indicates that Ames solution may be used as a suitable transport media for colorimetric detection. As shown in FIG. 29E, an experiment of directly mixing of a solid phase of SP and Ames solution showed that the solid phase of SP is not very soluble in the Ames solution and the solution remained yellow.
  • Butterfield's solution was tested for colorimetric detection.
  • Butterfield's solution comprises enzymatic digest of casein (l.OOg/L], monopotassium phosphate (0.04g/L], Polysorbate 80 (20mL/L], and the pH value of the solution was adjusted to 7.3 ⁇ 0.3.
  • a solution comprising Butterfield's solution and SP solutions showed a yellow color (FIG. 30A].
  • a solution comprising Butterfield's solution, SP and disodium succinate solutions showed a red- purple color (FIG. 30B]. This observation, consistent with those standard colorimetric detections (FIG.
  • Letheen Broth was tested for colorimetric detection.
  • Letheen broths are highly nutritious medias containing Lecithin and Tween® 80 for neutralizing quaternary ammonium compounds. These media may be modifications of the AOAC formulae.
  • Letheen Broth may be used for determining the phenol coefficient of quaternary compounds.
  • a typical composition of Letheen Broth may include peptone from meat 20.0(g/liter ⁇ ; peptone from casein 5.0 (g/liter ⁇ ; meat extract 5.0 (g/liter] yeast extract 2.0 (g/liter] sodium chloride 5.0 (g/liter] lecithin 0.7(g/liter]; sodium bisulfite 0.1 (g/liter ⁇ .
  • Letheen Broth may be prepared as the following procedure: Suspend 37.8 g and 5 ml of Tween® 80 in 1 liter of distilled or purified water until evenly dispersed; heat, if necessary, with repeated stirring to dissolve completely and autoclave at 121 °C for 15 min; maintain pH of 7.2 ⁇ 0.2 at 25 °C.
  • the as-prepared broth is turbid and yellowish-brown following incubation for 18 - 48 hours at 35 - 37 °C.
  • the Letheen Broth includes enzymatic digest of animal tissue (10 g/1], beef extract (5 g/L], sodium chloride (5 g/ L], Tween 80 (Polysorbate 80)(5 mL), Lecithin (0.7 g), with a final pH of 7.0 ⁇ 0.2 at 25°C.
  • a solution comprising Letheen Broth and SP solutions showed a yellow color (FIG. 31A ⁇ . Contrastingly, a solution comprising Letheen Broth, SP and disodium succinate solutions showed a red color (FIG. 31B ⁇ .
  • FIG. 31E An experiment of directly mixing of a solid phase of SP and Letheen Broth showed that the solid phase of SP is not very soluble in the Letheen Broth.
  • UTM-RT was tested for colorimetric detection.
  • UTM-RT comprises modified Hank's balanced salt solution, gelatin and bovine serum albumin as stabilizers, sucrose, glutamic acid and 4-(2-hydroxyethyl ⁇ - lpiperazineethanesulfonic acid (HEPES ⁇ .
  • HEPES ⁇ 4-(2-hydroxyethyl ⁇ - lpiperazineethanesulfonic acid
  • the presence of buffered salts in the medium protects pathogens that are sensitive to pH changes.
  • Gelatin and bovine serum albumin are source of nutrition to support viability of fastidious bacteria during storage and transport.
  • Sucrose aids in the preservation of viruses and chlamydiae when specimens are frozen for prolonged storage.
  • Antimicrobial agents are incorporated to minimize commensal bacterial and fungal contamination. Phenol red is added to act as a pH indicator.
  • a solution comprising UTM-RT and SP solutions showed a yellow color (FIG. 32A ⁇ . Contrastingly, a solution comprising UTM-RT, SP and disodium succinate solutions showed a red-peach color (FIG. 32B ⁇ . This observation, consistent with those standard colorimetric detections (FIG. 32C-32D ⁇ , indicates that UTM-RT may be used as a suitable transport media for colorimetric detection. As shown in FIG. 32E, an experiment of directly mixing of a solid phase of SP and UTM-RT showed that the solid phase of SP is not very soluble in the UTM-RT.
  • Elastic Polymers For Colorimetric Detection the chemicals for colorimetric detection include 3.556g Acrylamide, 0.0786g Calcium Chloride, 0.00213g N,N Methylene bisacrylamide, 6mg 4,4 Azobis(4cyanovaleric acid], 10ml DI Water, A separate polyacrylic acid solution includes 3ml DI Water, 0.350ml Acrylic Acid and 6 mg initiator. Both the polymer and polyacrylic acid solution were sonicated for mixing. The polyacrylic acid solution was then polymerized with UV. The resulting polyacrylic acid mixture (linearly polymerized] was added to the polymer mixture to form the elastic polymer solution.
  • the elastic hydrogel appears to be colorless. After the addition of all the chemicals of NIPs, the elastic hydrogel shows a yellow color (FIGS. 33D-33F ⁇ . Upon further addition of an analyte of disodium succinate, the elastic hydrogel appears to be pink. Further, it appears that the elastic properties of the hydrogel were maintained after the addition of NIP chemicals (FIGS. 33E-33F] and the analyte of disodium succinate (FIGS. 33H-33I ⁇ . These observations demonstrate that colorimetric detection of the present invention may be easily combined with elastic polymers and elastic hydrogels. The pink elastic hydrogel after the addition of the SP solution indicates the formation of MIPs as discussed above.
  • SP solutions Consisting of SP and a Solvent for Colorimetric Detection of ATP.
  • Solution comprises lmg SP and 20mL Dimethyl Sulfoxide (DMSO ⁇ . lmg of SP was dissolved in DMSO and sonicated for about 10 minutes. The resultant solution was blue.
  • SP solution samples were divided into control and test samples. 0.5mL of the solutions was added to plastic cups for illustration purposes. 0.5mL of water was added to the control samples. 0.5mL of ATP solution was added to the test samples. Note that SP and HEMA were used as a primary control sample where no water or ATP solution was added.
  • UV-Vis data also supports the color changes.
  • the yellow line represents the primary control sample of HEMA and SP at Img/mL in the absence of water.
  • the pink line represents the secondary control sample of HEMA and SP and DI Water in a 1 to 1 ratio.
  • the Green line represents the test sample with HEMA and SP and ATP solution in a 1 to 1 ratio. This observation corresponds with the proposed merocyanine (MC] isomers associated with the "chromic" properties of the SP molecule.
  • FIGS. 36A-36B highlight the transition of the peaks after ATP was added.
  • the shift in the peaks are consistent with a color change driven by the isomerization of SP ⁇ ->MC ⁇ ->MC'.
  • Previously Applicants associated the yellow color observed with the SP form of the molecule, and the Red color with that of the Merocyanine (MC] form of the molecule. It is possible that we are also looking at multiple forms of the MC isomer.
  • SP solutions Comprising SP, a Solvent and polymers for Colorimetric Detection of ATP.
  • Polymer Solution In one 20mL vial, the following was added.
  • SP and HEMA is first sonicated for lOmin. 20mL the SP+HEMA solution was added to the polymer components weighed out in the first vial, and then shaken and sonicated. The resultant solution was purple.
  • Solid Polymers In one 20mL vial, the following was added.
  • SP and HEMA is first sonicated for lOmin. 20mL of PBS was added to the SP+HEMA solution, and then shaken. Vial 2 was then added to Vial 1. The resultant solution was pink. 2mL of the respective polymer solution samples were polymerized by UV and cut into smaller pieces. ATP solution is used for testing is at a concentration of 20mg/mL DI Water
  • FIGS. 38A-38B color changes of the polymer pieces were observed before and after PBS was added to the polymers on the left (NIP], and after an ATP solution was added to the polymers on the right (NIP-tested with ATP].
  • FIGS. 39A-39E are a set of pictures showing time-dependent color changes of solid NIP polymers as shown in FIGS. 38A-38B.
  • the samples in the left lane are MIP solid polymers
  • the samples in the center lane are NIP solid polymers in the absence of ATP
  • the samples in the right lane are NIP solid polymers in the presence of ATP.
  • the top samples are corresponding polymer solutions before polymerization.
  • the NIP solid polymers or corresponding solutions turn yellow within 1 minute after the addition of ATP solutions.
  • FIG. 40 is a picture showing the color changes of NIP polymer solutions in the presence of different concentration of ATP.
  • the ATP concentrations were 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 mg/mL for the NIP polymer solutions from left to right. Color of the NIP polymer solutions shifts from purple/pink to lighter pink to yellow as ATP concentration increases.
  • FIG. 41 is a graph showing UV-Vis spectra of the corresponding NIP polymer solutions as shown in FIG. 40. Peak shift and intensity change can be associated with isomerization of SP.
  • FIG. 42 is a picture showing the color changes of SP solutions in dimethyl sulfoxide (DMSO] after addition of water or ammonium nitrate solutions.
  • SP solutions in DMSO show a blue color (left ⁇ . After addition of DI water into the SP solution in DMSO, the solutions turn pink (center ⁇ . After addition of an ammonium nitrate solution into the SP solution in DMSO, the solution turns yellow (right ⁇ .
  • a SP solution in DMSO was produced by dissolving suitable amount of SP in DMSO.
  • Ammonium nitrate solutions used in the experiments have a concentration of 10 mg/mL in DI water.
  • FIG. 43 is a graph showing UV-Vis spectra of the solutions as shown in FIG. 42.
  • the light blue line represents a primary control sample of SP solution in DMSO at lmg/mL in the absence of water.
  • the green line represents a secondary control sample of SP solution in DMSO and DI Water in a 1 to 1 ratio.
  • the dark blue line represents the sample of a SP solution in DMSO and an ammonium nitrate solution in a 1 to 1 ratio. Peak shift and intensity change can be associated with isomerization of SP.
  • FIGS. 44A-44B are a set of pictures and graphs showing ammonium nitrate detection by using SP solutions. After addition of DI water into the SP solution in DMSO, the solutions turn pink (left, FIG. 44A ⁇ . After addition of an ammonium nitrate solution into the SP solution in DMSO, the solution turns yellow (right, FIG. 44A ⁇ .
  • FIG. 44B shows UV-Vis spectra of the SP solution in DMSO after addition of DI water (purple ⁇ and after addition of an ammonium nitrate solution (yellow ⁇ .
  • FIGS. 45A-45B is set of pictures and graphs showing ammonium nitrate detection by using polymer and SP solutions.
  • Polymer and SP solutions were produced using similar protocols as discussed above.
  • the polymer and SP solution in the absence of ammonium nitrate shows a pink color (left, FIG. 45A ⁇ .
  • the polymer and SP solution turns yellow (right, FIG. 45A ⁇ .
  • FIG. 45B shows UV-Vis spectra of solutions as shown in FIG. 45A.
  • the pink line represents a control sample of polymer and SP solution.
  • the yellow line represents the sample of polymer and SP solution after addition of an ammonium nitrate solution.
  • FIGS. 47A-47F are a set of pictures showing time-dependent color changes of solid NIP polymers as shown in FIG. 46.
  • the samples in the left lane are MIP solid polymers
  • the samples in the center lane are NIP solid polymers in the absence of ammonium nitrate
  • the samples in the right lane are NIP solid polymers in the presence of ammonium nitrate.
  • the NIP solid polymers can start turning yellow within 1 minutes after the addition of ammonium nitrate solutions.
  • FIG. 48 is a picture showing the color changes of SP solutions in DMSO in the presence of different concentration of ammonium nitrate.
  • the ammonium nitrate concentration changes from 0 to 14 ⁇ L ⁇ .
  • the ammonium nitrate concentration changes from 18 to 500 ⁇ L ⁇ .
  • the solutions in the front row shows a concentration-dependent color change, and the color changes from purple/pink to yellow as the concentration of ammonium nitrate increases.
  • FIG. 49 is a graph showing UV-Vis spectra of the solutions as shown in FIG. 42. Peak shift and intensity change can be associated with isomerization of SP. After addition of ammonium nitrate solutions, a new peak was observed at 470 nm.
  • Minimal Solvent Requirements and Fine Tuning Colorimetric Detection are SP molecule and a specific solventfs ⁇ .
  • the specific solventfs] may be any suitable solvent.
  • the specific solventfs] may also be a combination of two more suitable solvents and compounds.
  • the minimum solution required for detection of an analyte is a solvent and spiropyran molecule.
  • the solvents may include Hydroxyethyl Methacrylate (HEMA] and SP.
  • HEMA Hydroxyethyl Methacrylate
  • SP SP
  • the solution of SP in HEMA was purple in color.
  • ATP oxidized glutathione
  • the color changed to yellow.
  • water was added to HEMA and SP solution. The final color was dark pink/purple.
  • FIGS. 44A- 44B ⁇ Another example is the detection of ammonium nitrate (see FIGS. 44A- 44B ⁇ . Ammonium nitrate was adding directly into a solution of acetone and SP or acrylic acid and SP. The resultant color changed from blue to purple and red to yellow, respectively.
  • Fine tuning requires the selection of a solvent that is specific for the detection of a particular analyte. Applicants found that one particular solvent alone may not be sufficient for the detection of all analytes. A specific solvent may be found to detect a specific analyte.
  • Fine tuning may also be concentration-dependent, which may be specific to a particular solvent. Each solvent may yield a different color in the presence of SP, and the color may change with the various concentrations of SP in the solution.
  • Fine tuning may also include SP and solvent interaction with the actual analyte.
  • the color readout in the presence of the analyte may vary with the type and concentrations of the solvent. Generally, a specific solvent or a solvent combination, concentration-dependency of the solvent and the interaction between SP and the solvent may be required to optimize a color readout.
  • the idea of fine tuning the technology for specific analyte detection as it pertains to solvents being used may refer to two specific dependencies. One may be the actual solvent being used. The second may be changes in solution concentration.
  • Table 1 lists some of the solvents as they have been used for detection with the associated analyte. Table 1 is not all inclusive, but serves as an example of "fine tuning" solvents for analyte detection.
  • the color readout of a solution may be heavily concentration-dependent.
  • concentration of the SP molecule within solution has an effect on the color observed within the solution, and therefore the color readout observed during the detection of an analyte.
  • This phenomenon may also be solvent-dependent.
  • SP in Acetone at very low concentrations yields a colorless solution.
  • the solution is blue.
  • SP in DMF at low concentrations yielding a green color.
  • a blue color is observed.
  • this dependency also affects the time required for the color change.
  • the solution may take longer to change color in the presence of the analyte.
  • the readout may be faster, but the color may be washed out as it is too light. Due to this dynamic phenomenon, a specific solvent and SP concentration may be determined for each analyte to optimize the readout observed. It should also be mentioned that the color scale previously observed as analyte concentration increases may also be determined by the solvents used in the solutions.
  • a second part to fine tuning may include the addition of a second solvent, e.g., water. Due to the solvatochromic properties of the SP molecule, the addition of water to each of the solvents may have a major effect on the color of the final solution, as well as the color readout during detection. Taking these components and parameters into consideration, Applicants determine how a molecule can be detected by establishing the solution parameters needed to optimize the color change and color readout.
  • a second solvent e.g., water

Abstract

La présente invention concerne des dispositifs et des procédés associés destinés à indiquer la présence d'un analyte cible dans une sécrétion corporelle située sur la peau d'un sujet. L'analyte cible est de préférence une drogue illicite ou un métabolite de celle-ci.
PCT/US2014/063398 2013-10-31 2014-10-31 Dispositif de détection d'analyte et procédés d'utilisation de celui-ci WO2015066459A1 (fr)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106038080A (zh) * 2016-07-13 2016-10-26 广东茵茵股份有限公司 具备汗液参数检测功能的吸湿用品
WO2017132618A1 (fr) * 2016-01-27 2017-08-03 Undercover Colors, Inc. Appareil pouvant être porté pour détecter une substance cible dans un liquide
US20200093416A1 (en) * 2017-06-02 2020-03-26 Northwestern University Thin, soft, skin-mounted microfluidic networks for detection and analysis of targets of interest in sweat
EP3641644A4 (fr) * 2017-06-23 2021-07-28 Voyant Diagnostics, Inc. Système et procédé de diagnostic médical
CN115236230A (zh) * 2022-07-21 2022-10-25 司法鉴定科学研究院 一种毛发中野生乌羽玉致幻成分麦司卡林的检测方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5203327A (en) * 1988-09-08 1993-04-20 Sudor Partners Method and apparatus for determination of chemical species in body fluid
US6443892B1 (en) * 2000-09-28 2002-09-03 The United States Of America As Represented By The Secretary Of The Navy Sweat collecting device and methods for use and detection of tampering
US6585646B2 (en) * 2000-11-29 2003-07-01 Hermetic Diagnostics, Inc. Screening test and procedure using skin patches

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5203327A (en) * 1988-09-08 1993-04-20 Sudor Partners Method and apparatus for determination of chemical species in body fluid
US6443892B1 (en) * 2000-09-28 2002-09-03 The United States Of America As Represented By The Secretary Of The Navy Sweat collecting device and methods for use and detection of tampering
US6585646B2 (en) * 2000-11-29 2003-07-01 Hermetic Diagnostics, Inc. Screening test and procedure using skin patches

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017132618A1 (fr) * 2016-01-27 2017-08-03 Undercover Colors, Inc. Appareil pouvant être porté pour détecter une substance cible dans un liquide
WO2017132614A1 (fr) * 2016-01-27 2017-08-03 Undercover Colors, Inc. Appareil, système et procédé de détection de substance cible
US10016315B2 (en) 2016-01-27 2018-07-10 Undercover Colors, Inc. Wearable apparatus for detecting a target substance in a liquid
US10024873B2 (en) 2016-01-27 2018-07-17 Undercover Colors, Inc. Methods and apparatus for detecting compounds in liquids
CN108780076A (zh) * 2016-01-27 2018-11-09 隐蔽色彩股份有限公司 用于检测液体中目标物质的可穿戴装置
CN106038080A (zh) * 2016-07-13 2016-10-26 广东茵茵股份有限公司 具备汗液参数检测功能的吸湿用品
US20200093416A1 (en) * 2017-06-02 2020-03-26 Northwestern University Thin, soft, skin-mounted microfluidic networks for detection and analysis of targets of interest in sweat
EP3641644A4 (fr) * 2017-06-23 2021-07-28 Voyant Diagnostics, Inc. Système et procédé de diagnostic médical
US11397176B2 (en) 2017-06-23 2022-07-26 Everyplace Labs, Inc. Medical diagnostic system and method
CN115236230A (zh) * 2022-07-21 2022-10-25 司法鉴定科学研究院 一种毛发中野生乌羽玉致幻成分麦司卡林的检测方法
CN115236230B (zh) * 2022-07-21 2024-01-19 司法鉴定科学研究院 一种毛发中野生乌羽玉致幻成分麦司卡林的检测方法

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