Reactive Surfaces Ltd. LLP
`Ex. 1031 (Rozzell Attachment L)
`Reactive Surfaces Ltd. LLP v. Toyota Motor Corp.
`IPR2016-01914
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`WO 2007/017701 A1
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`— before the expiration of the time limit for amending the
`claims and to be republished in the event of receipt of
`amendments
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`For two—letter codes and other abbreviations, refer to the ”Guid—
`ance Notes on Codes andAbbreviations” appearing at the begin—
`ning of each regular issue of the PCT Gazette.
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`WO 2007/017701
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`PCT/GB2006/050234
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`Fingerprint Analysis Using Mass Spectrometry
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`Field of the invention
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`The invention relates to a method for determining the presence of a residue within a
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`fingerprint using mass spectrometric techniques.
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`Background to the invention
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`Latent fingerprints contain numerous compounds such as naturally occurring compounds
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`from the body, e.g. cholesterol, squalene and fatty acids, [1-3] or compounds which may
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`be left on the latent fingerprint from a contact, e.g. cocaine or other drugs of abuse. Of the
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`studies to date with this objective, use has been made of Raman spectroscopy [4,5].
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`In
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`these studies, difficulty was observed in visually locating the drugs of abuse in order to
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`perform the analysis and the method was also relatively insensitive and relatively non-
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`specific. The most common method for the interrogation of latent fingerprints is Gas
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`Chromatography— Mass Spectrometry (GC—MS).
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`It has previously been shown that
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`residues from latent fingerprints can be extracted into a solvent and analysed by GC—MS
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`[2,3]. Such compounds include squalene and cholesterol, however levels of these on
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`latent fingerprints vary, not only between individuals, but between times for the same
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`individual [3]. GC—MS has also been used to detect contact residues, such as cocaine
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`from spiked fingerprints, with a limit of detection of approx 300 pg [6] and for the detection
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`of drugs of abuse and metabolites from commercial sweat patches down to ng per patch
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`levels [7] and from saliva [8]. However all of the above-mentioned methods require
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`complicated extraction procedures to be undertaken prior to analysis.
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`Matrix-assisted
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`laser desorption/ionization mass
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`spectrometry time-of-flight mass
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`spectrometry (MALDI-TOF-MS) was developed in late 1980s by Karas and Hillenkamp,
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`and has become established as a technique for the analysis and accurate molecular
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`weight determination of large macromolecules such as proteins, polysaccharides, nucleic
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`acids and synthetic polymers with high mass accuracy, and extreme sensitivity. MALDI is
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`a "soft" ionization process that produces minimum fragmentation, and in which the energy
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`from the laser
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`is spent
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`in volatilizing the matrix rather
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`than in degrading the
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`macromolecule. MALDI-TOF-MS has not been considered in the field of identifying
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`residues present on latent fingerprints. MALDI-TOF-MS is termed surface assisted laser
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`desorption/ionisation (SALDI) when graphite,
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`titanium dioxide or silica are used as
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`suspension matrices for MALDI [9, 11].
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`Summary of the invention
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`The present invention discloses various materials that can be used in detection and/or
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`imaging of fingerprints. These materials are typically also capable of acting as matrix-
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`agents in various mass spectrometric techniques. Thus, these materials have particular
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`properties enabling them to carry out such a “dual-purpose” role.
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`Thus, in one aspect of the present invention, there is provided a method of determining
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`the presence of a residue in a fingerprint, said method comprising:
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`i) applying to the fingerprint a particulate matter comprising a material which (1) is
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`capable of acting as a matrix agent or material in a matrix-assisted mass spectrometric
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`technique; and (2) aids detection and/or imaging of a fingerprint, to form a particle-applied
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`fingerprint; and then;
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`ii) subjecting the material
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`forming the particulate-applied fingerprint
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`to mass
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`spectrometry so as to detect the presence or absence of the residue.
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`In one embodiment, the method comprises use of materials, for example, metals, metal
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`oxides, metal nitrides and carbon,
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`that can be used (1) as agents for visualizing
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`fingerprints, either by themselves or combined with or embedded within a vehicle, for
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`example a silica vehicle, and (2) as a matrix for interrogating (analyzing) prints using a
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`matrix-assisted mass spectrometric technique. The mass spectrometry technique is used
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`to identify the presence or absence of substances such as one or more endogenous
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`compounds or metabolites, exogenous compounds or metabolites and/or contact residues
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`which the fingerprint includes.
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`In one embodiment, the mass spectrometric technique is
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`selected from (1) MALDI-TOF-MS and (2) SALDl-TOF-MS and (3) combinations thereof.
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`The fingerprint to which the method is applied may be a print which has been lifted from a
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`surface using lifting tape.
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`In one embodiment, the particulate matter is hydrophobic to facilitate application and
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`contacting of the particulate matter to a fingerprint.
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`It will be understood that the term “fingerprint” includes reference to a partial print and/or
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`to prints of other body parts and that, for example, a portion of a fingerprint to which the
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`particulate matter has been applied may be subjected to mass spectrometry. Typically, a
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`print is lifted from its underlying surface prior to the application of mass spectrometry, and
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`the term “fingerprint” accordingly includes lifted fingerprints.
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`ln embodiments,
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`the
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`fingerprint is lifted prior to application of the particulate matter.
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`It is contemplated that the
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`invention includes methods in which step ii) comprises subjecting particulate matter which
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`has acquired analyte from the print to the mass spectrometry.
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`It is further contemplated
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`that the invention includes methods in which step (ii) comprises subjecting both fingerprint
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`material and particulate matter to the mass spectrometry.
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`It will be understood that the terms “sample” and/or “analyte” in the context of the present
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`invention can be taken to mean a print, a sample taken from a print and/or a residue
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`present on or included in the print.
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`In some methods of the invention, a fingerprint is lifted from a surface and applied with the
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`particulate matter (processing agent), whether before or after lifting and the lifted
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`fingerprint (at least, material comprised in the print) is then placed in a mass spectrometry
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`apparatus.
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`In other methods, a print is made directly on a sample support and, after
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`application of the processing agent to the print, the sample support is placed in the mass
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`spectrometric apparatus.
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`In one embodiment,
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`the method further comprises locating and/or visualizing the
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`fingerprint and interrogation of the print using the above-described mass spectrometry
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`technique, for example, MALDl-TOF-MS and/or SALDl-TOF-MS.
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`According to an aspect of the invention, there is provided a method of determining the
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`presence of a residue within a fingerprint located on a surface, said method comprising
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`the steps of:
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`i) applying to the fingerprint a processing agent, for example, particulate matter
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`comprising a material selected from a metal, metal oxide, metal nitride, a carbon particle
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`and combinations thereof;
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`ii) subjecting the fingerprint
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`to mass spectrometry to detect the presence or
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`absence of said residue.
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`As previously stated,
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`therefore, the invention relates to detection, optionally including
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`quantification, of residues within fingerprints. The term “residue” refers to any material
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`which it is desired to detect for, in particular, pre-selected compounds. The residue is, or
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`may be, within i.e.
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`included in a fingerprint, that is the material which constitutes the
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`fingerprint contains, or is suspected of containing or may contain the residue.
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`In one embodiment, the particulate matter is hydrophobic.
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`In one class of methods, the
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`particulate matter may be used in combination with other matrix agents and/or other
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`fingerprint detecting agents.
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`The present invention provides a method which enables detection of (1) an endogenously
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`produced substance e.g. proteins,
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`lipids, DNA, peptides and/or endogenously derived
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`metabolites which is present as a residue included in a fingerprint; (2) an exogenous
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`compound or metabolite which is present as a residue included in a fingerprint; and/or (3)
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`a contact residue which is present on or within a fingerprint. Examples of the types of
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`residues are discussed later, but include for example (1) squalene and cholesterol; (2)
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`cocaine and its metabolites and nicotine and its metabolite and (3) ballistic residues from
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`e.g. firearms and/or explosives, residues from handling drugs of abuse (narcotics) e.g.
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`cocaine.
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`In one embodiment, the method also enables the detection of contact residues which are
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`co-deposited onto surfaces together with endogenously derived components (i.e.
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`endogenous metabolites and/or exogenous metabolites).
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`The method described herein typically does not require the complicated extraction
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`procedures prior to analysis associated with the prior art techniques and further provides
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`lower limits of detection.
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`One class of methods seeks to determine the presence or absence of a predetermined
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`substance.
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`In this case, the mass spectrum is examined for the presence of one or more
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`peaks associated with this known substance. Another class of methods seeks to identify
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`one or more substances in a print by comparing peaks in the mass spectrum with a
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`database or library of peaks. Both classes of methods may be performed in combination.
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`In one embodiment, the method comprises detection and/or identification of a residue
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`included in a fingerprint which has been directly deposited onto a MALDI-TOF-MS or
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`SALDI-TOF-MS sample support prior to application of the particulate matter.
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`In one
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`alternative embodiment, the method comprises detecting the presence or absence of a
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`residue within a fingerprint which has been lifted using a lifting means, for example lifting
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`tape, from a surface. The surface may be the site of deposition of the fingerprint at, for
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`example, a crime scene. The method may comprise contacting the lifted fingerprint with a
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`MALDI-TOF-MS or SALDI-TOF-MS sample support prior to application of the particulate
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`matter.
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`The method may alternatively comprise contacting the fingerprint with the particulate
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`matter prior to application of lifting means, for example, lifting tape, to the fingerprint.
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`In
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`one embodiment, a surface is first dusted to locate a latent print. This is followed by lifting
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`the dusted print with lifting tape. The print, located on the lifting tape is then applied to a
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`MALDI or SALDI target plate (sample support) prior to MS analysis.
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`Typically, a MALDI-TOF-MS or SALDI-TOF-MS sample support tends to be a plate, for
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`example, a stainless steel plate, which is designed to fit into an MS system. The plate
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`may comprise a well or plurality of wells to which a sample (Le. a fingerprint, for example
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`a lifted fingerprint) is added.
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`In one embodiment, the prints are semi-solid deposits
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`present on a sticky surface of the lifting tape.
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`The method may be employed qualitatively to determine the presence or absence of a
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`residue and/or quantitatively to determine the amount of a residue. Furthermore, in one
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`embodiment,
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`the method can be used to visualize or image the fingerprint.
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`This
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`visualization or imaging is important as it enables “owner” of the fingerprint
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`to be
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`identified.
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`In order to aid in the visualisation of the fingerprint, preferably the particle
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`further comprises a dye, for example a fluorescent or coloured dye. Appropriate dyes will
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`be known to those skilled in the art, but may, for example include, rhodamine 6G.
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`In one aspect, the present invention also provides use of particulate matter comprising
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`metal, metal oxide and/or carbon for the detection of a residue or residues on a fingerprint
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`in a matrix-assisted mass spectrometry technique.
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`In one aspect of the present invention, there is provided use of a matrix-assisted mass
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`spectrometry technique in the identification of
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`residues included in a fingerprint.
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`Particularly, there is provided use of a MALDl-TOF-MS and/or SALDl-TOF-MS technique
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`for detecting a residue in a fingerprint. The residue may be an endogenous residue (e.g.
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`an endogenous substance or metabolite for example squalene or an exogenous
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`metabolite, for example a drug or a drug metabolite), and/or a “contact” residue, for
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`example, ballistic residues from, typically, explosives or firearms.
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`In one embodiment, a
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`MALDl-TOF-MS technique and/or SALDl-TOF-MS technique may be used with fingerprint
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`imaging agents which can be used to detect (particularly visualize) fingerprints. Examples
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`of conventional fingerprint agents include aluminum, Magneta Flake and commercial white
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`powder.
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`In one embodiment, suitable matrix agents are used in the MALDl-TOF-MS/
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`SALDl-TOF-MS technique to assist in the desorption/ionization process. Examples of
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`conventional matrix agents include 2,5-Dihydroxybenzoic acid (DHB or DHBA) and q-
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`cyano-4-hydroxy cinnamic acid (o-CHCA).
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`In an alternative embodiment,
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`the use of MALDl-TOF-MS and/or SALDl-TOF-MS
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`comprises use of a particulate matter which comprises a material selected from (1) metal;
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`(2) metal oxide (3) metal nitride and (4) carbon as a matrix agent. The particulate matter
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`may further comprise additional features which are described herein. The particulate
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`matter typically may also be used as an agent for detecting and/or imaging a fingerprint.
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`In one embodiment, the fingerprint has a residue within it (i.e.
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`included in it.).
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`In one
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`embodiment, the particulate matter may be used in combination with other matrix agents
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`or materials (also known sometimes as matrix assistance agents and/or matrix enhancing
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`agents).
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`In one embodiment, the use may further comprise identification of the residue.
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`Detailed Description of the Invention
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`Throughout the description and claims of this specification, the words “comprise” and
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`“contain” and variations of the words, for example “comprising” and “comprises”, means
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`“including but not limited to”, and is not intended to (and does not) exclude other moieties,
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`additives, components, integers or steps.
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`Throughout the description and claims of this specification, the singular encompasses the
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`plural unless the context otherwise requires.
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`In particular, where the indefinite article is
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`used,
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`the specification is to be understood as contemplating plurality as well as
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`singularity, unless the context requires othenlvise.
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`Features, integers, characteristics, compounds, chemical moieties or groups described in
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`conjunction with a particular aspect, embodiment or example of the invention are to be
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`understood to be applicable to any other aspect, embodiment or example described
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`herein unless incompatible therewith.
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`Throughout this disclosure, various aspects of this invention can be presented in a range
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`format.
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`It should be understood that
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`the description in range format
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`is merely for
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`convenience and brevity and should not be construed as an inflexible limitation on the
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`scope of the invention. Accordingly, the description of a range should be considered to
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`have specifically disclosed all the possible subranges as well as individual numerical
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`values within that range. For example, description of a range such as from 1 to 6 should
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`be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4,
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`from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within
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`that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the
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`range.
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`It should also be understood that description of a number of ranges should be
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`considered to have specifically disclosed a combination of end points.
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`Method
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`The present invention relates to use of mass spectrometry in fingerprint residue analysis.
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`Specifically,
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`the present
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`invention relates to the use of a matrix-assisted mass
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`spectrometry technique,
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`for example, MALDl-TOF-MS and/or SALDl-TOF-MS in the
`
`analysis of fingerprint residues.
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`As stated previously, the term “residue” refers to any material which it is desired to detect
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`for, in particular, pre-selected compounds. The residue is, or may be, within i.e. included
`
`in a fingerprint,
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`that
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`is,
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`the material which constitutes the fingerprint contains, or is
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`suspected of containing or may contain the residue.
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`In one aspect of the present invention, there is provided a method for determining the
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`presence of a residue on a fingerprint, the method comprising (i) applying a particulate
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`matter to a fingerprint, the particulate matter having the following properties: (1) it
`
`is
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`suitable for use as a matrix agent in a matrix-assisted mass spectrometry technique, and
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`(2) it is suitable for use as an agent for detecting and/ or imaging a fingerprint; and (ii)
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`subjecting the fingerprint to mass spectrometry to detect the presence or absence of said
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`residue.
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`Preferably, the particulate matter is hydrophobic. The invention further includes methods
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`for
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`analyzing
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`a
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`hydrophobic
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`substrate
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`by matrix-assisted mass
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`spectrometry,
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`characterized in that
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`the matrix is hydrophobic.
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`In one embodiment,
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`the method
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`comprises, e.g. prior to step (ii), preparing at least one calibration standard for use in
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`calibrating the mass spectrometric technique.
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`In an embodiment, the method comprises
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`analyzing the outcome of step (ii) to e.g. determine whether a specific residue (e.g.
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`nicotine) is present in the fingerprint.
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`As stated above, the skilled person will understand that the term “fingerprint” includes
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`reference to a partial print and/or to prints of other body parts and that, for example, a
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`portion of a fingerprint to which the particulate matter has been applied may be subjected
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`to mass spectrometry. Typically, a fingerprint
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`is lifted from the surface prior to the
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`application of mass spectrometry, and the term “fingerprint” accordingly includes lifted
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`fingerprints.
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`ln embodiments, the fingerprint is lifted prior to application of the particulate
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`matter.
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`It is contemplated that the invention includes methods in which step ii) comprises
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`subjecting the particulate matter to the mass spectrometry.
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`It is further contemplated that
`
`the invention includes methods in which step (ii) comprises subjecting both the fingerprint
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`and the particulate matter to the mass spectrometry.
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`In one class of methods, step (i)
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`comprises immersing an article on which a print is or may be deposited in a liquid medium
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`comprising the particulate matter before being removed. The length of immersion is not
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`critical and may vary from about 15 minutes to about 12 hours or longer. The print may
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`then be lifted from the article using lifting tape.
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`It will be understood that the terms “sample” and/or “analyte” in the context of the present
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`invention can be taken to mean a fingerprint and/or a residue present on or within the
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`fingerprint, whether deposited directly or alternatively lifted from a surface using lifting
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`means, for example, lifting tape.
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`In one embodiment, the mass spectrometric technique is selected from MALDl-TOF-MS
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`and SALDl-TOF-MS.
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`In short, MALDl-TOF-MS requires mixing of samples with matrix
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`molecules and the application of the matrix material to a sample or the like. The MALDI-
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`target is introduced into the ion source of a mass spectrometer which is under a high
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`vacuum. A strong electrical
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`field is applied between the sample and the extraction
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`plate(s). A laser is fired onto the sample,
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`resulting in a desorption event due to
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`absorbance of the laser energy by the matrix molecules.
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`Thus, the invention comprises methods which utilize a material which is suitable as a
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`matrix material
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`in a MALDl-TOF-MS and/or SALDl-TOF-MS process. Without being
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`bound by scientific theory, it is considered that the particulate matter contains a material
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`which can absorb energy from the laser and transfer it to an analyte which is comprised in
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`the sample.
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`In the present invention, the analyte is typically a substance forming a
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`residue or residues within a fingerprint. The transfer of energy to the analyte results in
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`ionization of the analyte and acceleration through the mass analyzer. When MALDl-TOF-
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`MS is used in this way,
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`i.e. with transfer of ions to a sample (analyte),
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`it is known as
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`positive ion detection.
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`In an embodiment, the transfer of electrons may be from the analyte to the particulate
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`matter.
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`In this embodiment, the MALDl-TOF-MS (or SALDl-TOF-MS) is considered to be
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`running in a negative ionization mode.
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`If the sample (for example, the residue or suspected residue on or within the fingerprint)
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`is believed to have functional groups that readily accept a proton (H+) then positive ion
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`detection may be used.
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`If the sample (for example, the residue or suspected residue on
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`or in the fingerprint) is suspected to have functional groups that readily lose a proton then
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`negative ion detection is used.
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`The method may enable the presence of
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`(i) endogenous residues for example
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`endogenous metabolites and exogenous metabolites and (ii) contact residues included in
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`the fingerprint to be determined. The endogenous metabolites and “contact” residues
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`may be co-deposited within the fingerprint. The method may further enable identification
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`of the residue.
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`One class of methods may be used to detect and/or identify endogenous residues, for
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`example residues which have been produced as a result of metabolism of a substance by
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`a person’s body. The endogenous residues may include endogenous metabolites (e.g.
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`metabolites of molecules produced by the body) or exogenous metabolites (e.g.
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`metabolites of molecules ingested or transferred into the body and subsequently
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`metabolized by the body).
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`Other examples of endogenous residues which may be identified by the method include
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`for example endogenous substances (e.g. squalene, cholesterol, waxes and esters,
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`steroids e.g. estrogens and testosterone and markers of gender and health) which may be
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`secreted through skin pores and deposited with other chemicals within the fingerprint.
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`The method may also be used to detect
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`the metabolites and conjugates of the
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`aforementioned.
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`Examples of endogenous residues may also include exogenous
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`metabolites for example drug and their metabolites including drugs of abuse and their
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`metabolites, prescribed drugs and metabolites and compounds derived from dietary
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`sources or breakdown products of the same. The method could also be applied to the
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`proteomic or genomic analysis of the cells (e.g shed skin cells) or DNA respectively
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`located within the developed print. The method may also be used to detect other contact
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`residues for example, illegal drugs e.g. narcotics, explosive material, for example, material
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`used in bomb making processes, and residue from the use of a fire arm.
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`In a further aspect of the present invention, there is provided use of a MALDl-TOF-MS
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`and/or SALDl-TOF-MS technique for detecting and/or identifying a residue included in a
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`fingerprint. The residue may be an endogenous residue and/or a “contac ” residue, for
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`example, ballistic residues from, for example, explosives or firearms.
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`In one embodiment,
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`the residue may be a “contac” residue,
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`that
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`is to say, a residue which has been
`
`transferred to a person’s hand through contact with a substance and subsequently
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`transferred to a surface with a person’s fingerprint. The detection of such residues is of
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`particular interest to law enforcement agencies and may be prove to be a critical piece of
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`forensic evidence.
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`In one embodiment, a particulate matter which comprises a material
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`selected from metal, metal oxide, carbon and combinations thereof is used to contact the
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`fingerprint and act as a matrix in the MALDl-TOF-MS apparatus
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`In an alternative embodiment, the fingerprint may be contacted by a fingerprint developing
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`agent which can be used to detect fingerprints. Examples of conventional fingerprint
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`agents include aluminum, Magneta Flake and Commercial White. The method may then
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`typically use a MALDl-TOF-MS matrix agent for example DHBA (2,5-dihydroxybenzoic
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`acid) or o-cyano-4-hydroxy cinnamic acid (o-CHCA) or another matrix agent which may
`
`be e.g. the particulate matter as described herein.
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`The method of the present invention may be used to analyse a variety of residues which
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`may be found on a fingerprint. Thus,
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`in an embodiment, the method may be used to
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`develop and analyze latent fingerprints from smokers. It is well established that nicotine is
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`extensively metabolised to cotinine in vivo [22] and there is evidence that both nicotine
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`and cotinine are excreted together in sweat [8]. Figures 11 and 12 demonstrate that the
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`particulate matter as described herein, particularly, hydrophobic silica particles containing
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`a substance selected from metal, metal nitrides, metal oxides and carbon can be used as
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`developing agents to visualise latent fingerprints from smokers, and that such prints can
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`be analysed by MALDl-TOF-MS either directly on a suitable surface or following lifting
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`from the surface, to detect nicotine and/or its metabolites.
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`In one embodiment, the method described herein may be used to detect or determine
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`whether a person has handled or ingested drugs of abuse, for example, cocaine.
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`In other embodiments, the method described herein may be used as part of a personnel
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`screening to determine whether a person is, for example, a smoker or has taken drugs of
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`abuse for example cocaine.
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`Whilst the method described can be used to identify whether a particular residue is
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`present on or within a person’s fingerprint, the results are not to the highest level of
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`accuracy and may therefore be unacceptable currently to the standard required by for
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`example, criminal courts.
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`In this instance, e,g. if the method is being carried out for the
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`purpose of evidence for use in a court of law,
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`it may further comprise tandem mass
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`spectrometry, that is to say carrying out a further mass spectrometry technique, typically
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`to provide structural information for a compound present in a residue in a fingerprint.
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`Thus,
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`in one embodiment,
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`the method comprises subjecting a fingerprint and/or
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`particulate matter to MALDl-TOF-MS-MS and/or SALDl-TOF-MS-MS. MALDl-TOF-MS-
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`MS/ SALDl-TOF-MS-MS typically fragments specific sample ions
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`inside a mass
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`spectrometer and therefore provide further structural information about a residue. Thus,
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`in one embodiment, the method comprises identifying the resulting fragment ions. This
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`structural
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`information may be useful
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`in certain situations, for example, as mentioned
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`above as evidence in a court of law.
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`Method and uses described herein can be used in a variety of applications.
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`In one
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`embodiment, the methods and uses can be utilized as part of a personnel screening
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`process, for example by employees, to determine whether a person smokes or an abuser
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`of illegal drugs.
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`In an embodiment, the methods and uses can be utilized as part of a
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`“drug-testing” process in the field of, for example, professional or amateur sports.
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`In one embodiment, the method described herein may be used to detect residues, either
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`contact residues or endogenous residues which contain illegal or banned substances
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`and/or metabolites thereof. Particularly, the method may be used to test professional
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`and/or amateur sportspeople at sports events and/or random tests for the presence or
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`absence of banned substances in a residue taken from the person’s fingerprint. The
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`advantage of using the present method over currently used methods is that analysis is
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`performed directly on an individual’s own fingerprint and therefore substitution of a sample
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`is not possible as is the case with currently used methods of urine and saliva samples
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`followed by drug analysis screens.
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`The use of MALDl-TOF-MS and/or SALDl-TOF-MS to detect and/or identify a residue on
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`a fingerprint has not previously been considered and therefore the use of these
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`techniques to detect and/or identify a residue on a fingerprint forms part of the invention.
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`The use may comprise contacting of a particulate matter as described herein to a
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`fingerprint. Alternatively, the use may comprise contacting of a conventional fingerprint
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`detecting agent to a fingerprint.
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`In this embodiment, the use may further comprise use of
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`a matrix agent which is conventionally known in a mass spectrometry step. Alternatively,
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`the particulate matter as described herein can be used as a matrix agent in a mass
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`spectrometry step.
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`The methods and uses of the present invention may also be used at points of entry into a
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`country to test travelers for presence of residues which may suggest e.g.
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`illegal drug
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`handling or taking, handling of prohibited substances e.g. firearms and ballistics.
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`As described above, the methods can be used as forensic evidence of for example drug
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`or ballistic handling in a court, e.g. a criminal court.
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`Particulate Matter
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`The present invention utilizes particulate matter which can be contacted with a fingerprint
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`and which can subsequently be used in a mass spectrometry technique to determine
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`whether a residue is present on the fingerprint.
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`The particulate matter preferably comprises a material selected from metal, metal nitride,
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`metal oxide, carbon and combination thereof.
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`In one embodiment of the present
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`invention,
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`the particulate matter may comprise
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`hydrophobic silica particles. One class of hydrophobic silica particles is obtainable by the
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`following method:
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`A method (designated method A) for preparing hydrophobic silica particles is provided
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`which comprises reacting together in a single step a mixture of (1) silane ether monomers,
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`for example, a alkoxysilane and (2) organically substituted silane ether monomers, for
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`example a phenyl modified silicate, with a hydrolysing agent e.g. an alkali.
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`Thus, the method typically comprises the use of alkoxysilane monomers. The method may
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`comprise the use of tetraalkoxysilanes (abbreviated herein to TAOS). The TAOS’s are
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`particularly selected from TEOS (tetraethoxysilane) or TMOS (tetramethoxysilane).
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`In one embodiment, the mixture further comprises a water miscible solvent, for example,
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`ethanol, and also water. The method may be carried out at ambient temperature. The
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`duration of the reaction is not critical. The reaction between the TAOS monomers and
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`PTEOS monomers may be performed overnight or for an equivalent time period, that is to
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`say for between about 12 and about 18 hours. The length of the reaction has an effect on
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`the size of silica particles produced.
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`It is believed that the earlier a reaction is stopped,
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`the smaller the particles which are formed. Therefore, the reaction can be performed over
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`a period of less than 12 hours e.g. between 6 and 12 hours. The reaction may be
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`alternatively performed for longer than 18 hours.
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`If desired, the temperature may be
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`elevated (or reduced) and the duration of the reaction reduced (or increased).
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`The hydrolysing agent, typically an alkali, acts as a catalyst within the reaction. Preferably
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`this catalyst is a hydroxide, for example ammonium hydroxide. The catalyst may instead
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`be an acid. Examples of acids are mineral acids, e.g. hydrochloric acid. In this method,
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`the reaction comprises an acid induced hydrolysis.
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`The silane ether monomer, for example a TAOS, and the organically substituted silane
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`ether monomer, e.g. PTEOS monomers may be used,
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`for example,
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`in
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`ratios
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`(PTEOS:TAOS) of from 2:1 to 1:2 e.g. 4:3 to 3:4 and in particular 1.2:1, to 1:1.2.
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`In on