Mass spectrometry method of imprint analysis

FIELD: medicine.

SUBSTANCE: invention refers to medicine. A matrix mass spectrometry technique is used to detect the presence of a residue in an imprint. A powder containing hydrophobic particles of silicon dioxide, as well as a metal, metal nitride, a metal oxide or carbon is applied on the imprint. An average particle diametre of said metal, metal nitride, metal oxide or carbon introduced in the hydrophobic particles of silicon dioxide is 65 to 90 mcm or 400 to 500 nm preferentially. The investigated imprint is lifted from the application area with using a lifting tape and brought in contact with a sample substrate of a mass spectrometre, then the powder is applied on the imprint. The powder is magnetic or paramagnetic and additionally contains a fluorescent or pigmented dye molecule. The mass spectrometry is conducted with using a matrix specified in a group including MALDI-TOF-MS-MS or SALDI-TOF-MS-MS and their combinations. The residue represents an endogenous residue, particularly endogenous and/or exogenous metabolites and a contact residue. The endogenous metabolite represents squalene, while the exogenous metabolite is nicotine metabolite, e.g. cotinine, and the contact residue is the drug cocaine.

EFFECT: method allows executing the imprint residue analysis on a human imprint directly that eliminates it to be replaced.

21 cl, 14 dwg

 

The technical field

The present invention relates to a method of determining the presence of the residue in print, using the technique of mass spectrometry.

The level of technology

Latent fingerprints contain various compounds such as natural bodily connection, for example, cholesterol, squalene, and fatty acids [1-3], or compounds that may remain in the latent fingerprint by contact, for example, cocaine or other drugs. To examine these residues up to the present time have used spectroscopy Raman scattering [4, 5]. When conducting such analyses arises the difficulty associated with the difficulty of visual localization of the drug for analysis, and this method has low sensitivity and is relatively nonspecific. The most common way analysis of latent fingerprints is gas chromatography - mass spectrometry (GC-MS). Previously it was shown that the remains of latent fingerprints can be extracted in the solvent and analyzed using GC-MS [2, 3]. Such compounds include squalene and cholesterol, but their levels in the latent prints vary, not only between subjects (individuals), but also in time for the same subject [3]. GC-MS has also been used to detect contact the s residue, such as cocaine-rich prints, with a detection limit of about 300 mcg [6], and for the detection of drugs and metabolites from serial sweat stains, to nanogramme levels on a single spot of sweat [7]and saliva [8]. However, all mentioned here above methods require the use of complex procedures extraction of the earlier analysis.

Laser desorption/ionization using matrix time - of-flight mass spectrometry (MALDI-TOF-MS) was developed in the late 1980s researchers Karas and Hillenkamp, and was used as a technique of analysis and determine the exact molecular weight of large macromolecules, such as proteins, polysaccharides, nucleic acids and synthetic polymers, with high precision measurement of the mass and with very high sensitivity. MALDI is a soft ionization process, which creates minimal fragmentation, and in which energy from the laser is spent rather on evaporation of the matrix, and not on the destruction of macromolecules. However, MALDI-TOF-MS were not previously used to identify residues that are present in latent prints. MALDI-TOF-MS was carried out using surface laser desorption/ ionization ("SALDI"), when graphite, titanium dioxide or silicon dioxide is used as the matrix suspension for MALDI [9, 11].

The invention

In accordance with this from what Britanie offers a variety of materials, which can be used for detection and/or visualization of fingerprints. These materials typically can also be used as a matrix agent in various techniques of mass spectrometry. Thus, these materials have special properties that enable their use as materials "dual use".

In accordance with the first aspect of the present invention, it is proposed a method of determining the presence of the residue in print, and this method includes the following operations:

i) coating the fingerprint powder containing material that is capable of (1) to act as a matrix agent or material in the technique of mass spectrometry using matrix; and (2) to facilitate detection and/or imaging of the fingerprint to form the imprint of the coated particles; and then

ii) carrying out mass spectrometry of the material forming the imprint of the coated particles to detect the presence or absence of residue.

In accordance with one embodiments, the method involves the use of materials such as, for example, metals, metal oxides, metal nitrides, and carbon, which can be used (1) as agents for the visualization of fingerprints, by themselves or in combination with the carrier or with the introduction of a carrier, for example, the nose of the tel of silicon dioxide, and (2) as a matrix for the analysis of prints using the technique of mass spectrometry using matrix. The technique of mass spectrometry is used to determine the presence or absence of substances such as one or more endogenous compounds or metabolites of exogenous compounds or metabolites and/or contact residues that contains the imprint. In accordance with one variant of implementation, the technique of mass spectrometry is chosen from the group comprising (1) MALDI-TOF-MS and (2) " SALDI " -TOF-MS and (3) combinations thereof.

Imprint, to which is applied the proposed method may be a fingerprint lifted from the surface using lifting tape.

In accordance with one embodiments, the powder is hydrophobic, in order to facilitate the application of the powder on the fingerprint and the contact of the powder with it.

It should be borne in mind that the term "mark" includes a partial fingerprint and/or fingerprint image of the finger, and the other part of the body, and, for example, the area of the imprint, which was deposited powder may be subjected to mass spectrometry. Typically, a fingerprint lifted from the underlying surface of the previously conducting mass spectrometry, the term "fingerprint" includes such raised prints. In accordance with the variations in implementation, an imprint raise earlier application by osca. The present invention includes the ways in which the operation (ii) provides for mass spectrometry powder, which has acquired the analyte from the imprint. The present invention also includes methods in which the operation (ii) provides for mass spectrometry as a material of the imprint, and powder.

It should be borne in mind that the terms "sample" and/or "analyte" in the context of the present invention can mean an imprint, a sample of the fingerprint, and/or the residue present on the print or in print.

According to some here ways, the fingerprint lifted from the surface and put on him the powder (processing agent), before or after lifting, and then raised imprint (at least the material contained in the print) is placed in the device for mass spectrometry. In accordance with other methods, an imprint of doing (put) directly on the substrate of the sample, and, after application of the processing agent on the imprint substrate sample is placed in the device for mass spectrometry.

In accordance with one embodiments, the method further provides for localization and/or visualization of the fingerprint and the fingerprint analysis using the technique described here mass spectrometry, for example, MALDI-TOF-MS/or " SALDI " -TOF-MS.

In accordance with the first aspect of the present invention, it is proposed a method of determining the presence of residue in the imprint, located on the surface, and this method includes the following operations:

i) coating the print processing agent, such as a powder containing material selected from the group comprising metal particles, metal oxide, metal nitride, carbon, and combinations thereof; and

ii) carrying out mass spectrometry fingerprint to detect the presence or absence of the specified residue.

As mentioned here above, the present invention relates to the discovery, which may include quantitative determination of residues in the prints. The term "residue" refers to any material that you want to detect, and in particular, relates to a pre-selected compounds. The residue contains or may contain imprint, so that the material which forms the imprint of, contains or may contain residue.

In accordance with one embodiments, the powder is hydrophobic. In accordance with one class of methods, the powder can be used in conjunction with other matrix agents and/or with other agents detect a fingerprint.

In accordance with the present invention it is proposed a method, which provides the AET detection (1) endogenously produced substances, for example, proteins, lipids, DNA, peptides and/or endogenous derived metabolites, which are present as a residue contained in the imprint; (2) exogenous compound or metabolite that is present as a residue contained in the imprint; and/or (3) contact residue, which is present in the fingerprint or thumbprint. Examples of the types of residues are discussed here below, and they include, for example, (1) squalene and cholesterol; (2) cocaine and its metabolites and nicotine and its metabolite, and (3) ballistic residues, for example, from firearms and/or explosives and residues from drugs such as cocaine.

In accordance with one embodiments, the method also allows to detect contact residues, which were deposited on the surface together with endogenously produced compounds (i.e. endogenous metabolites and/or exogenous metabolites).

The described method typically does not require complicated procedures extraction of the earlier analysis, as in previously known methods, and also provides lower detection thresholds.

In one class of methods seek to determine the presence or absence of a given substance. In this case, investigate the mass spectrum for the presence of one or more peaks associated with a known substance. In another Klah is CE methods seek to identify one or more substances in the imprint, by comparing the peaks in the mass spectrum of a database or library of peaks. Both classes of methods can be implemented in combination with each other.

In accordance with one embodiments, the method involves the detection and/or identification of residue contained in the print that was directly deposited on a substrate (plate) sample MALDI-TOF-MS or " SALDI " -TOF-MS before applying the powder. In accordance with an alternative embodiment, the method involves detecting the presence or absence of the residue in print that was lifted from the surface using the lifting means, such as lifting tape. The surface may be the place deposits imprint, for example, at the crime scene. The method can include contacting the raised imprint with the substrate sample MALDI-TOF-MS or " SALDI " -TOF-MS, previously applying powder.

The alternative method may include contacting the fingerprint powder previously overlay on the fingerprint lifting means, such as lifting tape. In accordance with one embodiments, the first surface is sprinkled with powder, to localize the latent fingerprint. Then raise dusted with fingerprint powder using lifting tape. Imprint localized on lifting the ribbon, and then put in place the inu target (substrate samples) MALDI or " SALDI", followed by MS (mass spectrometry) analysis.

Typically, the substrate sample MALDI-TOF-MS or " SALDI " -TOF-MS is a plate, for example, a plate of stainless steel, which is used in the MS system. The plate may have a recess or multiple recesses, into which enter the sample (i.e. the fingerprint, such as raised print). In accordance with one variant of implementation, the prints are semi-solid deposits (sludge) on the sticky surface of the lifting tape.

The method can be used qualitatively to determine the presence or absence of residue and/or quantitatively to determine the amount of residue. Moreover, in accordance with one embodiments, the method can be used for visualization or imaging of the fingerprint. This visualization or imaging are important because they allow to identify the "owner" of the print. To facilitate visualization of the printout, the particles mainly contain a dye such as a fluorescent or colored (dyed) dye. Appropriate dyes known to specialists in this field, and the dye can be, for example, rhodamine 6G.

In accordance with the embodiment of the present invention provides the use of powder, which contains meta is l, the metal oxide and/or carbon, for detection of the remnant or residue on the paper, using the technique of mass spectrometry using matrix.

In accordance with the first aspect of the present invention, it is proposed the technique of mass spectrometry using matrix to identify residues contained in the imprint. In particular, it is proposed to use the technique of MALDI-TOF-MS and/or " SALDI " -TOF-MS for detection of the residue in print. The balance can be endogenous residue (for example, an endogenous substance or a metabolite, for example, it may be squalene, or exogenous metabolite, such as drug or metabolite drugs), and/or "contact" residue, for example, ballistic residues from explosives or firearms. In accordance with one variant of implementation, the technique of MALDI-TOF-MS and/or technique " SALDI " -TOF-MS can be used with agents visualization of fingerprint to detect (and in particular to visualize) imprints. As typical examples of agents (visualization) of the fingerprint, you can bring aluminum, Magneta Flake and trading a white powder. In accordance with one embodiments, a suitable matrix agents used in the technique of MALDI-TOF-MS/ " SALDI " -TOF-MS to facilitate desorption/ ionization. As typical examples of the matrix is hentov can lead 2,5-dihydroxy benzoic acid (DHB or DHBA) and α-cyano-4-hydroxyanisol acid (α-SNS).

In accordance with an alternative implementation, the use of MALDI-TOF-MS and/or " SALDI " -TOF-MS involves the use of powder, which contains a material selected from the group which includes (1) metal; (2) a metal oxide; (3) a nitride of the metal; and (4) carbon as a matrix agent. The powder may have the additional characteristics described here. The powder typically can also be used as an agent for detecting and/or imaging of a fingerprint.

In accordance with one variant of implementation, the fingerprint is contained in the residue. In accordance with one embodiments, the powder can be used in combination with other matrix agents or materials (sometimes referred to as agents of the support matrix and/or agents gain matrix).

In accordance with one variant of implementation, the use of advanced can identify the remainder.

Detailed description of the invention

In the following description of the invention and in the claims, the words "includes" and "include" and variations such as "includes" and "including"mean "includes, but without limitation" and do not serve to limit (or limit) the use of other shares, additives, components, integers or operations.

In the following description and the gain and in the claims, the singular includes the plural, unless there are special instructions otherwise. In particular, when the English text uses the indefinite article, in the description of the invention provides for the use of both the singular and the plural, unless otherwise evident from the context.

It should be borne in mind that the features, integers, characteristics, compounds, chemical components or groups described in accordance with a particular aspect, embodiment or example of the present invention is applicable in any described here another aspect, embodiment or example, if there is no incompatibility with him.

In the description of the present invention various aspects set forth in the sequence. However, it should be borne in mind that this sequence is shown for convenience only and are not restrictive. Thus, the range of descriptions includes all possible subranges and individual rooms.. for Example, the description in the range from 1 to 6 includes specific subranges from 1 to 3, 1 to 4, from 1 to 5, 2 to 4, from 2 to 6, from 3 to 6 and so on, as well as individual numbers within that range, for example, 1, 2, 3, 4, 5 and 6. This applies regardless of the width of the range. It should also be borne in mind that the ranges in the description include the JV is nificence disclosed combination of endpoints.

Way

The present invention relates to the use of mass spectrometry in (at) the analysis of the remainder of the print. In particular, the present invention relates to the use of technology mass spectrometry using matrix, for example, MALDI-TOF-MS and/or " SALDI " -TOF-MS in the analysis of the fingerprint.

As mentioned here above, the term "residue" refers to any material that you want to detect, and in particular, to a pre-specified compounds. The remainder is or may be located within the footprint, i.e. the material which forms the imprint of, contains or may contain residue.

In accordance with one aspect of the present invention, it is proposed a method of determining the presence of residue on the print, and the method involves (i) applying the powder on the fingerprint, and the powder has the following properties: (1) it is suitable for use as a matrix agent in the technique of mass spectrometry using matrix, and (2) it is suitable for use as an agent for the detection and/or visualization of the imprint; and (ii) carrying out mass spectrometry of the fingerprint to detect the presence or absence of the specified residue.

Mainly, the powder is hydrophobic. In accordance with the present invention also provides methods for the analysis of hydrophobic substrates by means of mass-SP is chromefree using matrix characterized in that the matrix is hydrophobic. In accordance with one embodiments, the method involves, for example, earlier surgery (ii)preparation of at least one calibration standard for use in the calibration technique of mass spectrometry. In accordance with one embodiments, the method involves the analysis of results of operations (ii), for example, to determine that a particular residue (e.g., nicotine) is present in the fingerprint.

As mentioned here above, the experts will easily understand that the term "mark" includes a partial fingerprint and/or prints not fingers, and other parts of the body, and, for example, the area of the imprint, on which was applied the powder, can be used to perform mass spectrometry. Typically, a fingerprint lifted from the surface prior to mass spectrometry, the term "fingerprint" accordingly includes raised prints. In accordance with a variant implementation, the fingerprint raise before applying the powder. In accordance with the present invention suggests ways in which the operation (ii) provides for selection of powder for mass spectrometry. In accordance with the present invention also provides methods in which the operation (ii) provides for mass SPECT is hometree as fingerprint, and powder. In one class of methods of operation (i) involves immersing the object on which there is or may have an imprint, in a liquid medium containing the powder. The time of immersion is not critical and can range approximately from 15 minutes to 12 hours or more. The fingerprint can then be lifted from the object using lifting tape.

It should be borne in mind that the terms "sample" and/or "analyte" in the context of the present invention can mean the fingerprint, and/or residue that is present in the imprint or contained in a print deposited directly or, alternatively, raised from the surface using the lifting means, for example, lifting tape.

In accordance with one variant of implementation, the technique of mass spectrometry is chosen from the group which consists of MALDI-TOF-MS and " SALDI " -TOF-MS. Briefly, MALDI-TOF-MS requires mixing of the samples with matrix molecules and deposition of matrix material on the sample. The MALDI target is introduced into the ion source of the mass spectrometer, which is under high vacuum. A strong electric field forming between the sample and the extraction plate (plates). The laser beam is directed onto the sample, resulting in desorption due to the absorption of laser energy by the molecules of the matrix.

Thus, in accordance with the present invention offers the proposed methods, using a material that can be used as a matrix material in the technique of MALDI-TOF-MS and/or " SALDI " -TOF-MS. Not wishing to be bound to any specific theory, it is still possible to say that the powder contains material which can absorb the laser energy and transfer it to the analyte contained in the sample. In accordance with the present invention, the analyte is typically a substance that forms a residue or residues in the imprint. The energy transfer to the analyte leads to ionization of the analyte and to accelerate through the mass spectrometer. When MALDI-TOF-MS used in this way, i.e. with the transfer of ions in the sample (analyte)is called a positive ion detection.

In accordance with the embodiment, it can result in the transfer of electrons from the analyte in the powder. In this embodiment, MALDI-TOF-MS (or " SALDI " -TOF-MS) is in the negative ionization mode.

If you think that the sample (for example, balance or suspected balance or in print) has functional groups that are readily accept a proton (H+), then can be used for positive ion detection. If you think that the sample (for example, balance or suspected balance or in print) has a functional group that readily lose a proton, then can be used for negative ion detection.

One class of methods can be used to detect and/or identify endogenous residues, for example, residues that have been obtained in the metabolism of nutrients by the body. Endogenous residues may contain endogenous metabolites (e.g., metabolites are molecules produced by the body) or exogenous metabolites (e.g., metabolites molecules, swallowed, or moved into the body and then metabolized by the body).

Other examples of endogenous residues that can be identified using the proposed method are, for example, endogenous substances (e.g., squalene, cholesterol, waxes and esters, steroids, such as estrogen and testosterone and markers of gender and health), which may be released through the pores of the skin and deposited together with other chemicals in the imprint. The method can also be used to detect metabolites and conjugates of the above-mentioned substances. Examples of endogenous residues are also exogenous metabolites, such as drug and its metabolites, and the drug is its metabolites, prescription drugs and metabolites and compounds derived from power sources or their decay products. The method can also be applied to proteomic (proteomic or genomic analysis of cells (e.g. skin cells) or DNA, respectively, are shown in the imprint. The method can also be used to detect other contact residues, such as drugs, explosive material, for example of the material used in the manufacture of bombs, and residue from the use of firearms.

In accordance with another aspect of the present invention, it is proposed technique (MALDI-TOF-MS and/or " SALDI " -TOF-MS for the detection and/or identification of a residue, which is contained in the imprint. The balance can be endogenous residue and/or "contact" residue, for example, ballistic residues, for example, from explosives or firearms. In accordance with one variant of implementation, the balance can be "contact" balance, i.e. the balance that was transferred into the hands of man through contact with the substance and then transferred to the surface due to fingerprints. Detection of such residues is of particular interest to law enforcement and can serve as critical evidence in judicial review. In accordance with one of the option is in the implementation, the powder, which contains a material selected from the group comprising metal, metal oxide, carbon, and combinations thereof, used for contact with the imprint, and it acts as a matrix in MALDI-TOF-MS device.

In accordance with an alternative embodiment, the fingerprint may be introduced into contact with the agent manifestations of the printout, which is used to detect fingerprints. As examples of suitable agents manifestations of print, you can bring aluminum, Magneta Flake and trading a white powder. The method then typically used MALDI-TOF-MS matrix agent, for example, DHBA (2,5-dihydroxy benzoic acid) or α-cyano-4-hydroxyanisol acid (α-SNS), or other matrix agent, which may be, for example, described herein powder.

The method in accordance with the present invention can be used to analyze a number of residues that can be found on the imprint. Thus, in accordance with one embodiments, the method can be used for the display and analysis of latent prints left by smokers. Firmly established that nicotine is highly metabolised in cotinine in vivo (in a living organism) [22], and it is known that nicotine and cotinine are excreted in sweat [8]. Figure 11 and 12 shows that this powder, and in particular, hydrophobic particles is of oxida silicon, contains a substance selected from the group comprising metal, metal nitrides, metal oxides and carbon, can be used as developers for the visualization of latent fingerprints obtained from smokers, and these fingerprints can be analyzed using MALDI-TOF-MS, directly on a suitable surface or after lifting from the surface to detect nicotine and/or its metabolites.

In accordance with one of the embodiments described herein may be used to detect or determine what people had to deal with drugs or used drugs, such as cocaine. In accordance with other variants of implementation, the described method can be used as part of a personal screening to determine that the person is, for example, a smoker or takes drugs, such as cocaine.

Although the described method can be used to identify the presence of specific residue on or in a print person, these results do not have the highest level of accuracy and can therefore be unacceptable in accordance with the standards used, for example, in criminal courts. In this case, for example, when the method is carried out to obtain evidence for trial, he may additionally contain the substance of the conduct of tandem mass spectrometry, that is, the implementation of the additional techniques of mass spectrometry, typically, to obtain structural information about the compounds present in the residue in the printout.

Thus, in accordance with one embodiments, the method involves carrying out MALDI-TOF-MS-MS and/or " SALDI " -TOF-MS-MS for print and/or powder. When carrying out MALDI-TOF-MS-MS/ " SALDI " -TOF-MS-MS typical fragment ions specific probes inside the mass spectrometer and thereby gain additional structural information concerning the rest. Thus, in accordance with one embodiments, the method involves identification of the obtained fragment ions. This structural information can be used in some situations, for example, as evidence in court.

Method and uses described herein may be used in various applications. In accordance with one embodiments, the methods and uses can be used as part of the process of personal screening for employees, for example, to determine whether the person smokes or takes drugs. In accordance with one embodiments, the methods and uses can be used as part of process validation for drugs", for example, in the field of professional or Amateur is one sport.

In accordance with one of the embodiments described herein may be used to detect contact residues or endogenous residues, which contain a prohibited substance and/or their metabolites. In particular, the method can be used to test athletes in professional or Amateur sport and/or random testing for the presence or absence of prohibited substances in the residue, taken from the imprint of a man. The advantage of the proposed method in comparison with currently used methods is that the analysis is performed directly on the imprint of a man and therefore replacement of the sample is impossible, in contrast to currently used methods, which examine samples of urine and saliva.

Device MALDI-TOF-MS and/or " SALDI " -TOF-MS has not previously been used for the detection and/or identification of a residue on the print, so the use of these techniques for the detection and/or identification of a residue on the print is part of the present invention. This may include contacting the powder with the imprint, as mentioned here above. Alternatively, use may include contacting an ordinary agent detection with imprint. In accordance with this embodiment, it can additionally be COI is used a matrix agent, which is usually used in mass spectrometry.

The alternative described here previously, the powder can be used as a matrix agent when performing mass spectrometry.

The methods and uses according to the present invention can also be used at border crossings for travelers checks for the presence of residues, which may indicate the use of drugs or treatment (contact) with them, and contact with firearms and ammunition.

As mentioned here above, the proposed methods can be used as evidence in court, for example, as evidence of contact with drugs or ammunition in a criminal court.

Powder

In accordance with the present invention it is proposed to use a powder that is introduced into contact with the imprint, which can then be used in the technique of mass spectrometry to determine the presence of residual imprint.

Powder mainly contains material selected from the group comprising metal, metal nitride, metal oxide, carbon, and combinations thereof.

In accordance with one embodiments of the present invention, the powder may contain hydrophobic particles of silicon dioxide. One class of hydrophobic particles of silicon dioxide receive according the following way:

Proposed method (denoted as method A) preparation of hydrophobic particles of silicon dioxide, which provides entry into the reaction in a single operation a mixture of (1) monomer ester silane, for example, alkoxysilane, and (2) organic substitution of the ester monomers of silane, for example, modified with phenyl silicate, agent hydrolysis, for example, alkali.

Thus, the method typically involves the use of monomers of alkoxysilane. The method may involve the use of tetraalkoxysilane (TAOS). TAOS choose from TEOS (tetraethoxysilane) or TMOS (tetramethoxysilane).

In accordance with one embodiments, the mixture further comprises mixing with water, the solvent, for example ethanol, and water. The method can be carried out at ambient temperature. The duration of the reaction is not critical. The reaction between the monomers TAOS and monomers PTEOS may occur during the night or during the equivalent period of time that is approximately 12 to 18 hours. The reaction time affects the size of the resulting particles of silicon dioxide. It can be assumed that the sooner will stop the reaction, the smaller will be formed particles. Therefore, the reaction should be carried out within a period of less than 12 hours, for example, during the period from 6 to 12 hours. eacce alternative may last more than 18 hours. Optionally, the temperature may be increased (or reduced) and the reaction time is reduced (or increased).

The agent hydrolysis, which is typically the alkali acts as a catalyst during the reaction. Mainly, this catalyst is a hydroxide, such as ammonium hydroxide. The catalyst may instead be acid. As examples of suitable acids can lead inorganic acid, for example hydrochloric acid. In this way the reaction is a excited by acid hydrolysis.

The monomer ester silane, such as TAOS, and organically substituted monomer ester silane, for example, PTEOS, can be used, for example, when the relations (PTEOS:TAOS) from 2:1 to 1:2, for example, from 4:3 to 3:4, and in particular from 1.2:1 to 1:1.2. In one class of methods, the ratio is at least about 1:1, for example, to 1:5, for example, 1:2. In one class of methods, the ratio of the PTEOS:TAOS is preferably 1:1 by volume. It should be borne in mind that when replacing one or both TAOS and PTEOS other reagents can be used similar relationship.

Hydrophobic particles of silicon dioxide, obtained by the described method are essentially the nanoparticles may have an average diameter of approximately 200 nm to 900 nm, and typically from approximately 300 nm to 800 nm, in particular from 400 nm to 500 nm. These nanocatalysts can be processed, to form microparticles, which can be considered fused nanoparticles. Microparticles can be obtained using a method which, for example, includes the following operations:

(i) centrifuging the suspension of particles;

ii) translation suspension of hydrophobic particles of silicon dioxide in the aqueous phase;

iii) extraction of sediment from the aqueous phase into the organic phase;

iv) evaporation of the organic phase; and

(v) grinding and sieving the product obtained in operation (iv).

The organic phase is primarily organic solvent which is non-polar or low polarity. The organic phase can be dichloromethane or another organic solvent, for example, alkanes, such as hexane, toluene, ethyl acetate, chloroform and diethyl ether.

Alternatively, the hydrophobic particles of silicon dioxide can be obtained from a reaction product containing hydrophobic nanoparticles of silicon dioxide, using a method which includes the following operations:

(a) centrifuging the reaction product;

(b) washing the reaction product fluid.

The method may include repeating the operations (a) and (b) many times. Mainly, the liquid is a mixture of the solvent with water, and typically water-organic solvent mixture. Typically, organic is rastvoritele is ethanol. Mainly, the initial liquid contains a mixture of water and organic solvent in the ratio of 60 (water):40 (solvent) to 40:60, by volume. In accordance with other variants of implementation, the solvent may be, for example, dimethylformamide, n-propanol or ISO-propanol.

Typically, the proportion of solvent in the mixture increases between the initial leaching (b) and final flushing of sediments. To obtain microparticles, which are fused nanoparticles, prepared slurry is dried. Then the microparticles can be sifted. After sieving, the particles are ready for use on print in as developing agent, for example, in the operation (i) the proposed method for detection of the residue in print.

Microparticles can be considered as aggregates of smaller nanoparticles of silicon dioxide. In accordance with this embodiment, the microparticles are quite large, allowing them to effectively capture using facial masks and, therefore, does not swallow. Thus, in accordance with one embodiment, the microparticles of silicon dioxide have an average diameter of at least 10 μm, and typically at least 20 μm. Typically, the microparticles have an average diameter of about 30 to 90 μm. In accordance with some of the options for implementation, the microparticles have an average is th diameter of about 45-65 μm or approximately from 65 to 90 μm. In particular, the powder containing microparticles may be a dry powder.

In one class of methods, the powder contains hydrophobic nanoparticles of silicon dioxide. Hydrophobic nanoparticles of silicon dioxide can be selected using a method which involves centrifugation of the reaction product of method a and his suspension in a mixture of water and solvent. A mixture of water and solvent in the first mixture and preferably is a mixture of 50:50. The method may further include removing the reaction product from the first mixture of water and solvent, centrifuging it and the suspension of his second mixture of water and solvent. Mainly, the second a mixture of water and solvent is similar to the first mixture, the proportion of solvent and water. The solvent in a mixture of water and solvent is, for example, mixed with water, the solvent, for example ethanol. Alternatively, it may be used dimethylformamide, n-propanol or ISO-propanol.

The operation of the suspension of the reaction product in a mixture of water and solvent can be repeated many times. Mainly, the composition of a mixture of water and solvent change in order to increase the proportion of solvent in the mixture during repeated operations of the suspension. Primarily, the method involves, in the operation, the suspension of the reaction product in a mixture of water and solvent, which contains about 0% water and 100% of the solvent. The total number of operations suspension typically ranges from 3 to 10, for example, 4, 5, 6, 7, 8 or 9. Typically, after each operation of the suspension, except for the final operation of the suspension, the suspension centrifuged. The nanoparticles can be stored in the final ethanol suspension. It should be borne in mind that the centrifugation represents only one exemplary method for separating nanoparticles from a mixture of water and solvent, so it is possible to use other suitable technology selection nanoparticles.

In accordance with one embodiments, the powder contains hydrophobic nanoparticles of silicon dioxide. One class of powder is suspended hydrophobic nanoparticles of silicon dioxide in the fluid. The fluid may be ethanol aqueous suspension. Alternatively, instead of ethanol in suspension can be used and other organic solvents, such as dimethylformamide, n-propanol or ISO-propanol.

The physical nature and size of the particles forming the powder, can be determined using SEM and TEM microscopes. The particles have the form of amorphous particles of silicon dioxide, which is usually used as non-stick additive for various food products, as well as antipr the Garn additive and as an excipient in the pharmaceutical industry in the preparation of various medicines and vitamins [12].

Thus, in accordance with the preferred embodiment, the powder, which contains nanoparticles, applied to the imprint or to the surface in an appropriate liquid medium. Typically, the liquid medium is a mixture of water component with the solvent. The solvent can be mixed with water, the solvent. In accordance with one embodiments, the aqueous component is water. The solvent may be, for example, mixed with water, the solvent, i.e. a solvent that is 100% miscible with water in all proportions. In accordance with one embodiments, the solvent is ethanol. The ratio of water to solvent is in the range approximately from 99.9:0.1 to 96:4. The solvent content mostly does not exceed approximately 4%, as higher levels of solvent can lead to the dissolution of prints or reducing the resolution. Mostly use at least trace amounts of solvent, so that the nanoparticles remained discrete and do not stick together to form aggregates.

Alternatively, the powder containing hydrophobic particles of silicon dioxide, can be obtained using conventional methods (see, for example, the publication of Taras et al., NanoSci. Nanotech. 2002. Vol.2. No.3/4 pp.405-409. E.R.Menzel, S.M. Savoy, S.J.Ulvick, K.H.Cheng, R.H.Murdock and M.R.Sdduth, Photoluminescent Semiconductor Nanocrystals for Fingerprint Detection, Journal of Forensic Sciences (1999) 545-551; and E.R.Menzel, M. Takatsu, R.H.Murdock, K.Bouldin and K.H.Cheng, Photoluminescent CdS/Dendrimer Nanocomposites for Fingerprint Detection, Journal of Forensic Sciences (2000) 770-773).

In accordance with one embodiments, the powder contains hydrophobic particles of silicon dioxide, which are introduced dye. In accordance with one embodiments, the dye, which entered into particles is, for example, a colored or fluorescent dye. As examples of suitable colorants can cause (but not limited to, fluorescein derivatives, e.g., Oregon Green, Tokyo Green, SNAFL and carboxylato the fluorescein, rhodamine (for example, rhodamine b and rhodamine 6G) and their analogues, thiazole orange, oxazin perchlorate, methylene blue, basic yellow 40, basic red 28, and crystal violet and its analogues. Not wishing to be bound to any specific scientific theory, it is still possible to believe that the dyes which are positively charged, for example, rhodamine, may be better put into particles, when the method is used PTEOS than dyes, which contain anionic or cationic group, such as carboxyl groups. As examples of other dyes that may be used in accordance with the present invention, it is possible to cause the dyes, which have a planar aromatic substructure and put the Ino charged functional group (for example, ethidium bromide and other built-in DNA agents).

Particles mainly can be magnetic or paramagnetic. For example, magnetic particles can be easily poured on the prints, using a magnetic probe or other suitable tool. Thus, in accordance with the preferred embodiment of the present invention, magnetic or paramagnetic subunit injected into the hydrophobic particles of silicon dioxide. In accordance with the embodiment of the present invention, the particles are magnetized, for example, magnetic or paramagnetic. Magnetic or paramagnetic particles can contain any magnetic or paramagnetic component, for example, metals, metal nitrides, metal oxides and carbon. As examples of magnetic metals can lead to iron, while the metal oxides can be magnetic and hematite.

In accordance with another preferred embodiment of the present invention, carbon is a carbon black, carbon nanoparticles, fullerene compound or graphite, as well as their analogues. Fullerene compound contains at least 60 carbon atoms (for example, C60). Mainly, carbon has the form of carbon nanoparticles. Carbon nanoparticles can be in the form of, for example the EP, carbon nanotubes (derived or not derived). Carbon nanotubes can be nanotubes with multiple walls or one wall.

In accordance with one embodiments, the metal oxide is chosen from the group which consists of titanium dioxide (TiO2), magnetite, hematite, and combinations thereof. In accordance with one embodiments, the metal is chosen from the group comprising aluminum, iron, and combinations thereof. However, it should be borne in mind that in an alternative implementation options, specialists in this field can use other metal oxides and/or nitrides of metals, which contribute to the process of desorption/ ionization MALDI-TOF-MS and/or " SALDI " -TOF-MS. Similarly, professionals in this field can be used in powder other metals and/or forms of carbon, which contribute to the ionization process. Metal, metal oxide, metal nitride or carbon can be introduced into the powder particles. The powder particles generally have an average diameter of ≤100 μm, for example, the diameter of ≤1 µm. In accordance with one embodiments, the particles have an average diameter of approximately 10 nm to 100 μm.

The hydrophobicity of the particles of silicon dioxide enhances adhesion of particles with imprint. Thus, in accordance with one embodiments, metal, metal oxide and carbon, e.g. the Yat and/or inserted into the hydrophobic particles of silicon dioxide.

In the ways in which they use the fingerprint powder and imprint enter into contact with each other. It should be borne in mind that the term "fingerprint" in this context can mean, for example, a fingerprint deposited on the surface, or, alternatively, "indirect" imprint, which was lifted from the surface using conventional lifting means, for example, using lifting tape.

Powder coating can be performed by using a magnetic probe, and in this embodiment, the powder is magnetic or paramagnetic. It is safer because it reduces the exposure of the powder to the person who can breathe. Alternatively, the object on which the imprint may be immersed in a liquid (i.e. in a suspension of nanoparticles). The time of immersion is not critical and can range approximately from 15 minutes to 12 hours or more.

In accordance with one embodiments, the powder contains hydrophobic particles of silicon dioxide, which contain magnetic particles, as mentioned here above.

One class of powder contains hydrophobic particles of silicon dioxide. Particles can be nanoparticles or microparticles, or a combination of both. In accordance with one embodiments, the microparticles of silicon dioxide which have an average diameter of at least 10 μm, and typically at least 20 μm. Typically, the microparticles have an average diameter of about 30 to 90 μm. In accordance with some of the options for implementation, the microparticles have an average diameter of from approximately 45-65 μm 65 to 90 microns.

Suppose that the nanoparticles that have an average diameter of approximately 200 nm to 900 nm, can be used in the proposed methods.

Mainly, the nanoparticles have an average diameter of approximately from 400 to 500 nm. However, I believe that in the proposed method can be used powder containing nanoparticles with an average diameter of approximately 200 nm to 900 nm, for example, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850 or 900 nm.

The term "average diameter" refers to the average diameter of the particles, typically formed by methods in accordance with the present invention. The term "average" is a statistical term, which is obtained by dividing the sum of all the measured diameters on the number of particles used in such measurements. The diameters of the nanoparticles determined from SEM images using scales (rulers), and the diameters of the microparticles was determined using a combination of the size of the Sith, measurements of particle size distribution and SEM images. One of the ways to determine the average diameter is determined using the device Malvern Mastersizer (firms who Malvern Instruments Ltd.).

In accordance with one embodiments, the powder may contain a mixture of (1) hydrophobic particles of silicon dioxide, as described here above, and (2) magnetic or paramagnetic particles, such as particles of iron.

In accordance with one embodiments, the powder contains hydrophobic particles of silicon dioxide, which additionally contain a molecule that helps with visualization and/or imaging of the fingerprint. In accordance with one embodiments, a hydrophobic particle of silicon dioxide contains a molecule of the dye. As examples of dyes can lead rhodamine, for example, rhodamine 6G and its derivatives. The operation of the rendering of the print can be performed using various known techniques. For example, can be used optical methods, including scanner with UV search light beam, an optical scanner with a planar scan fluorescence scanner and the scanner in the UV and visible region of the spectrum.

Examples

Variants of the present invention will be described below by way of example, with reference to the accompanying drawings.

Figure 1 shows the results of fluorescent scanning of fingerprints deposited on a glass slide and raised with him by using rhodamine 6G (Rh.6G) as contact agent (λ 543 nm, λm 590 nm).

On figa p is the accumulated latent fingerprints on a glass slide. The top and bottom prints with rhodamine 6G, in the center of the blank control.

On fig.1b shows fingerprints figa shown using powder "Sunderland White" (hydrophobic particles of silicon dioxide was put into them TiO2).

On figs shows the balance (traces) from prints fig.1b after their ascent using a commercially available tape for lifting fingerprints.

On fig.1d shown raised prints fig.1b on commercially available lifting the tape.

Figure 2 shows mass spectra of rhodamine 6G, precipitated from ethanol (higher), and rhodamine 6G from the raised imprint (lower), both the spectrum is analyzed on the ribbon MALDI (below).

Figure 3 shows the system response MALDI-TOP-MS particles of silicon dioxide was put into them, carbon black, in the presence and absence of cocaine hydrochloride.

Figure 4 shows the comparison matrix material for the detection of cocaine enriched prints are printed directly on the metal plate MALDI-TOF-MS using 2,5-dihydroxy benzoic acid (DHB) at a concentration of 10 mg/ml, and the upper curve was obtained using hydrophobic magnetic particles of silicon dioxide.

On figa comparison matrix material for the detection of cocaine enriched prints are printed directly on metal PLA is Tina MALDI-TOF-MS. The lower curve was obtained using DHB at a concentration of 10 mg/ml, and the upper curve was obtained using hydrophobic magnetic particles of silicon dioxide.

On fig.4b shows the spectral intensity at m/z 304.5 for fingerprints deposited on the metal plates of targets and sprinkled three powders in the presence and absence of cocaine.

Figure 5 shows mass spectra lifted latent prints, topped with hydrophobic particles of silicon dioxide entered Tio2to detect contact with cocaine. Positive contact is shown in the top curve, and the lack of contact with cocaine is shown on the lower curve.

Figure 6 shows a photograph of a single deepening/ single cell on the surface having the recesses 96 of the MALDI plate, located in the mass spectrometer, where you can see the detail of the ledge (light bars) raised imprint sprinkled hydrophobic particles of silicon dioxide entered Tio2before analysis using MALDI-TOF-MS.

7 shows the relative intensity at m/z 433.55 (molecular ion for squalene plus adduct of sodium) for the lifted prints sprinkled 10 formulations of hydrophobic particles (1-10), and for controls (11 and 12). Observed no peaks at m/z 386 due to cholesterol.

On Fig shown consider the performance communications intensity at m/z 433.55 (squalene, weight formula 410.72, plus adduct of sodium (22.99)for standards of squalene in the presence of eight formulations of hydrophobic powders (1-8) and two controls (9, 10).

Figure 9 shows the relative intensity at m/z 386.37 (weight formula 386.65 for cholesterol) standards for cholesterol squalene in the presence of eight formulations of hydrophobic powders (1-8) and two controls (9, 10).

Figure 10(a) shows the mass spectra MALDI-TOF-MS for the standard of squalene.

Figure 10(b) shows the mass spectra MALDI-TOF-MS for standards cholesterol.

Figure 11 shows the results of MALDI-TOF-MS analysis of latent fingerprints for non-smoker (FR) on the stainless steel plate and latent prints, enriched with 10 μl of a solution containing 1 mcg/ml of a mixture of nicotine (RMM 163) and cotinine (RRM 176). Bottom-up:

a) - enriched latent prints, not sprinkled hydrophobic particles or processed DHB;

b) - enriched latent prints processed DHB;

c) - enriched latent prints sprinkled hydrophobic particles;

d) is not enriched latent fingerprint sprinkled hydrophobic particles.

On Fig shows MALDI-TOF-MS of latent fingerprints for a former smoker (24 hours after the last cigarette), stainless steel plate, sprinkled hydrophobic particles:

(a) the m/z range 155-180, b) m/z is the range 180-205, C) m/z range 205-450.

On Fig shows MALDI-TOF-MS of latent prints for smoker, sprinkled with powder. Peaks at 163, 148, 133, 119, 105, 91, 84 and 79 are specific to nicotine and are the clearest evidence of his presence.

On Fig shows MALDI-TOF-MS of latent prints for smoker, sprinkled with powder. Peaks at 177, 161, 147, 135, 119, 105, 97, 91 and 79 are characteristic cotinine and serve as clear evidence of his presence.

Materials and methods

In this study, we used the scanner Tecan LS300 company Tecan UK., Reading, Berkshire, as well as system Kratos Axima-CFR MALDI-TOF-MS, Shimadzu Biotech, Manchester, UK) with the plates of the target Shimadzu. The mass matrix used 2,5-dihydroxy benzoic acid (DHB) (10 mg/ml in a solution of 50:50 acetonitrile: deionized water [dH2O]). The suspension of carbon black was kindly provided by the company Cabot Corp., Cheshire UK. All other chemicals were purchased on the company Sigma-Aldrich, Dorset UK., including titanium dioxide in the form of anatase. Serial apadravya tools, brushes for prints, magnetic probes and repetitive lifting tape were obtained in the company of a Crime Scene Investigation Equipment Ltd., 10 (formerly known as K9 Scenes of Crime Ltd.) Northampton, UK.

Was prepared solution for system calibration MALDI-TOF-MS. There were prepared solutions of papaverine hydrochloride (10 mg/ ml in dH2O) and reserpine (5 mg/ ml in dimethylformamide, DMF). An aliquot of the shown solution of papaverine hydrochloride (100 µl) was mixed with an aliquot of a solution of reserpine (200 μl) and dH 2O (400 ml) and DMF (300 μl). Also was prepared solution of 2,5-dihydroxy benzoic acid (DHB, 10 mg/ ml) in 50:50 by volume acetonitrile: water. The final solution for calibration was prepared by mixing 10 µl of each solution DHB and mixtures of reserpine and papaverine. Aliquots (1 μl) of this solution were used in each experiment, and m/z for the molecular ions of the two standards were used to calibrate the system.

Preparation of hydrophobic particles of silicon dioxide was put into them material

This method is an adaptation of preparation blank nanoparticles based on silicon dioxide [12] and involves mixing 30 ml of ethanol, 5 ml dH2O, 2.5 ml of tetraethoxysilane and 2.5 ml of phenyltriethoxysilane in the centrifuge tube. To the resulting add 2 ml of ammonium hydroxide solution (28%) to initiate the formation of nanoparticles and solution spinning throughout the night. The resulting slurry (suspension) powder extracted again using a solution of methylene dichloride/ water or ethanol/ water (50:50 in each case). The suspension is centrifuged (e.g., 5 minutes at 3000 rpm). Remove the supernatant and add 10 ml dH2O and the same volume of dichloromethane. Suspension rotate an additional 10 minutes and then centrifuged again. Remove the aqueous upper layer solution and add the LNO add aliquots of water and dichloromethane. This process of washing and centrifugation is repeated 4 times, until you need additional additive water: dichloromethane. After that, the dry particles from dichloromethane in a thermostat at 40°C.

Dry particles are crushed in a mortar and pestle and sieved to the desired size. Hydrophobic particles sift manually through the brass test sieves with bronze mesh (firm Endecot Ltd., London UK). The fraction of particles that are used in this trial have a size below approximately 63 μm. For particle sizing using the analyzer of the type Malvern Mastersizer, Malvern Instruments Ltd., Malvern, UK).

To obtain particles containing titanium dioxide, 25 mg titanium dioxide is injected into the centrifuge tube, then add the reagents of silanization. To obtain particles containing carbon black (CB), add 5 ml of 1:2-1:100 solution of suspension of carbon black in water solution predecessor instead of TiO2. To obtain magnetic particles using magnetite powder in accordance with the published method [13, 14], and then injected 5 ml of this suspension in water solution predecessor instead of dH2O.

Demonstration of the fact that the contact remains effectively raised from latent fingerprints

The tip of the finger was placed in a solution of rhodamine 6G (Rh 6G; 100 ál/ml in tO). The finger moved (soon) for evaporation of excess tO RAS is the first application fingerprint on a clean glass slide. Were applied three times, with 2 Rh 6G and 1 blank control. The prints were examined using scanner Tecan LS300 (543 nm λex, λem 590 nm, increased 120). Then prints sprinkled hydrophobic microparticles of silicon dioxide, which contain put into them, titanium dioxide, previously visualization under the same scanning conditions. After that, the prints were raised with the use of serial lifting tape (11.5×6.5 cm), and then scanned as a residue on a glass slide and lifting the tape.

Experiment to demonstrate MALDI-TOF-MS detection of raised contact residue (rhodamine 6G)

In this experiment a few grains of powder Rh. 6G inflicted directly on the tip of your finger, previously applying fingerprint on a glass slide, which was produced similarly to the previously described, but without adding outrivaled funds (powder). The fingerprint was then raised using double-sided conductive tape and deposited on a metal target. A conductive tape, suitable for use in conjunction with MALDI-TOF, was kindly provided by the company Shimadzu Biotech. The target plate was then introduced into the system MALDI-TOF-MS and imprint investigated for the presence of Rh. 6G. Received MS found m/z for Rh 6G approximately 444, compared with the standard (0.1 ál solution 100 ág/ ml in ethanol), equivalent to 100 ng of dye deposited on a flat tape is.

Experiment to demonstrate the effectiveness of particles of silicon dioxide was put into them, carbon black as a reinforcing agent for " SALDI " -TOF-MS

They studied the response of the instrument to particles of silicon dioxide was put into them, carbon black, in the presence and absence of cocaine, which was inserted in the circular cell/ area on the surface of the metal plate target. All results were obtained with the triple repetition. Cocaine hydrochloride (1 μg in 1 μl of 1 mg/ml ethanol)) was administered in 2 sets of holes on the target. In a positive test, 1 ál of 10 mg/ml suspension of the nanoparticles to put them in carbon black (matrix) was injected into the cell. In another control test studied the response of the instrument for cocaine without matrix. In yet another screening test studied the response of the instrument matrix in the absence of cocaine, using the empty cells of a metallic target. Cells were dried in accordance with the previously described until analysis.

The detection of the contact residues of cocaine using MALDI-TOF-MS a Small amount (a few grains) of cocaine hydrochloride were deposited on the tip of your finger, when it is immersed in the drug. Then the finger was put in direct contact with clean metal plate target, resulting in a received mark on the surface of the target, covering many cells (pits)on the surface. Latent prints are then sprinkled hydrophobic particles of silicon dioxide was put into them magnetite, or sprinkled serial opudrivayuschee means, which were powder of aluminum, Magneta Flake™ and white powder, may contain titanium dioxide. Serial magnetic probe used for applying two magnetic powders and used the standard brush for applying non-magnetic powders. For evidence of cocaine on the print used serial DHB matrix was mixed 10 μl of 10 mg/ml 10 ál 10 mg/ml solution of cocaine hydrochloride, and 1 µl of this mixture was injected into the cell MALDI target. Metal target and then dried in a heater (firm Shimadzu/Kratos Instruments) and then introduced into the MS system.

Experiment to demonstrate the effectiveness of TiO2/PTEOS particles for detection of residues of cocaine on the lifted fingerprint

Empty and containing cocaine prints were applied to glass slides in accordance with previously described. The prints showed using TiO2/PTEOS particles, applied with a brush Zephyr. The prints were then raised with the use of serial lifting tape, which turned and put the print up on the plate of the target system MALDI. The prints were then analyzed using a mass spectrometer (MS) for the presence or absence of cocaine.

The detection holes the Wendy Erin and squalene from prints using the system MALDI-TOF-MS, using hydrophobic particles as a reinforcing matrix

The metal plate of the target was cleaned and dried. Then 12 of the fingerprint of the index finger of the right hand were applied to the metal plate of the target. The plate with the prints were then dried in an incubator at 37°C for a time of about 1.5 hours. Each print sprinkled then ten different formulations of hydrophobic particles, prepared as described here above. The agent entered into particles, indicated below in parentheses. In addition, some of the obtained powders (1, 5, 6, and 8) and then thoroughly mixed with the DHB to get 1% by weight of the composition. They were used to determine that the presence of this matrix improves the detection of squalene in the latent fingerprint after sanding, the analysis in the system MALDI-TOF-MS. There were used four of the composition of black powder, which have different proportions of carbon black in them, derived from the original synthesis. The numbers in brackets refer to the ratio of black carbon to the dH2O used in the original synthesis:

1. White powder (titanium dioxide); 1.0% by weight DHB; powder)

2. White powder (titanium dioxide)

3. Purple powder (crystal violet)

4. Black powder And (1:10)

5. Black powder (1:10, 1.0% by weight DHB; powder)

6. Purple powder (crystal violet; 1.0% by weight DHB; powder

7. Red fluorescent dye (rhodamine 6G)

8. Red fluorescent dye (rhodamine 6G; 1.0% by weight DHB; powder)

9. Black powder (1:5)

10. Black powder D (1:2)

Each of the first ten prints individually sprinkled respective powders 1-10 and the resulting prints were photographed using a digital camera (photos not shown). Then the prints are individually analyzed in the system MALDI-TOF-MS, including the imprint, which has not been processed outrivals tool or matrix (an imprint of 12), and imprint processed only dihydroxy benzoic acid (imprint 11).

Direct detection of cholesterol and squalene using system MALDI-TOF-MS, using hydrophobic particles as a reinforcing matrix

Solutions of cholesterol and squalene (both 2 mg/ ml) were prepared in 100% ethanol and 500 μl of aliquots of each solution were mixed with each other. 24 drops (each 0.5 μl) of this solution were applied to clean, dry metal target and left to dry for 2 hours. Then deepening were individually sprinkled each with eight hydrophobic powders (3 drops for each composition). The compositions were identical to those described in 2.8:

1. Red fluorescent powder And

2. Black powder And

3. Black powder

4. Purple powder And

5. Purple powder

6. rainy fluorescent powder

7. Black powder

8. Black powder D

Rubbing the powder produced in accordance with the previously described herein above, directly on the surface of the metal plate, each of the powders. There was also prepared a separate solution by mixing 500 μl of a solution of cholesterol/squalene with 500 μl of a solution of 10 mg/ml DHB (matrix). Three drops (0.5 µl) of this solution was applied to the metal target and left to dry in the air. Every drop (spot) on the metal of the target was analyzed using MALDI-TOF-MS and recorded peaks of cholesterol (m/z 386) and squalene (m/z 410) and their metal adducts.

Results

Demonstration of the fact that the contact remains effectively raised from latent fingerprints

Studied the effectiveness of the method of lifting the print and details tabs of the resulting prints. Rhodamine 6G was used as a model contact agent due to its high fluorescence. Thus, the received fingerprint with high fluorescence after exposure (upper and lower marks on figa), while the normal print (center figa) is not fluorescent when used scanning modes.

When hydrophobic particles of silicon dioxide were applied to three times, they became visually clear (not shown), and now all three attacat is as clearly visible by fluorescence scanning (fig.1b) due to the fluorescence of rhodamine, as well as due to scattering of light from particles of silicon dioxide, deposited on the imprint. After lifting fingerprints using lifting tape, small fluorescence is retained on the surface of glass slides (figs), indicating that the efficiency of the method of ascent. Scanning the surface of the tape now detects fluorescent fingerprints, suggesting that the raised prints and optimouse means remain intact during this process (fig.1d).

The results, shown in figure 1, confirm that the serial lifting the tape can be used to transfer the total volume of deposited material latent prints from slides on the tape. This is the first operation upon detection of any material in the latent fingerprint carried out using MALDI-TOF-MS.

MALDI-TOF-MS detection of raised contact residue (rhodamine 6G)

The successful results of the previous experiment suggest that the rhodamine raised from the surface of the object-glass of the microscope, can be detected using MALDI-TOF-MS. The obtained results are shown in figure 2. The upper curve corresponds to the raised print and has a peak at m/z 443 corresponding to Rh 6G. The bottom spectrum obtained directly from Rh 6G standard printed directly on the target MALDI. Similar forms peaks suggest the presence of the same compounds. Reduced weight on the top of the curve is probably caused by a slight lifting tape, used when applying the lifting tape on the plate of the target, which reduces the flight time of the ionized species.

Conductive tape should be well glued to the plane of the MALDI target, and the sample is applied on the top surface of the tape. This tape is relatively fragile and easily stretched, which can deform the original manifest imprint. This tape can hardly be used for lifting prints from slides microscope and for application to the target system MALDI best. It is impossible to align the tape on the surface due to its fragility and stickiness.

The results, shown in figure 2, suggests that the contact remains in the form of a powder, can be lifted from the surface and can be successfully analyzed using MALDI-TOF-MS.

The efficiency of the use of particles of silicon dioxide was put into them, carbon black as a reinforcing agent upon detection of cocaine using MALDI-TOF-MS

For demonstration (proof) that the results were obtained due to the presence of matrix and cocaine, but not the background signal were conducted additional analyses using known what's quantities of cocaine and matrix. The response of the magnetic particles of silicon dioxide in the presence and absence of cocaine were compared with the responses of the empty target and cocaine without any matrix.

A comparison was made between responses at m/z 304 particles of silicon dioxide was put into them, carbon black, in the presence and absence of cocaine, with empty responses of the target and cocaine without any DHB matrix. The intensity of the MS peaks due to cocaine is shown in figure 3. The presence of particles of silicon dioxide leads to 10-fold increased intensity of the peak compared to the peak for cocaine in the absence of these particles. There is good reproducibility (average intensity 90,275 and rsd 5.8%, n=3) compared with the corresponding results in the presence of DHB (medium intensity 56,552 and rsd 112%, n=3).

Was investigated the potential of the system " SALDI " using particles with put into them, carbon black, compared to a standard DHB matrix. Cocaine hydrochloride was used as a given analyte, since its identification by fingerprints is of interest to law enforcement, because it shows that left the imprint of the subject has been in contact with cocaine. Mass spectra of cocaine in the presence of the standard chemical matrix (DHB) and a new powder matrix shown in figure 4, for prints, printed directly on metal target system MALDI. Neither the deposits range corresponds to cocaine in the presence of DHB, while the upper spectrum corresponds to the cocaine in the presence of a magnetic PTEOS nanoparticles deposited using a magnetic probe. Both the spectrum are similar and correspond to the spectrum of cocaine, however, the response is clearly increased in the presence of magnetic particles. This suggests that the magnetic PTEOS particles contribute to the process of desorption of cocaine the same as the normal matrix. Magnetic PTEOS particles have potential for use as a powder to render the print as seen a good ledge on the developed prints (not shown).

Similar results were also obtained for the peaks of cocaine at 304.5 m/z in case of contact prints deposited on the metal plate target and sprinkled hydrophobic powders containing titanium dioxide, black carbon (1:10), and processed DHB or left without treatment (details not shown), and relative intensity, respectively 1,700, 10,000, 9,000, and 100 mV (fig.4b).

Efficiency of particles introduced Tio2upon detection of residues of cocaine on the lifted prints

Successful demonstration of the effectiveness of the process of the rise (Fig 1 and 2) and detection of cocaine from a latent fingerprint on the target system MALDI (figure 3-5) suggest that cocaine can be found on the lifted prints C is directly with the serial lifting tape. The results shown in figure 5. The lower curve is obtained for the raised imprint, which has not been in contact with cocaine, while the upper curve is obtained for the imprint, which has been in contact with cocaine. A clear peak is visible at the top of the curve at m/z 304, which corresponds to cocaine. In either case, the original prints were dusted hydrophobic particles of silicon dioxide containing TiO2. 7 shows the region of the target system MALDI-TOF containing the imprint enriched with cocaine. A similar pattern is observed for the negative control (not shown). The diameter of the recess is 3.4 mm, and there is a clear ridge (ridge detail of the print. The spectra of figure 6 were obtained using laser scanning areas inside cells, and the signals were averaged to obtain the spectra.

Detection of cholesterol and squalene from prints using the system MALDI - TOF-MS, using hydrophobic particles as a reinforcing matrix

In this experiment, 12 prints were applied directly on the target plate of stainless steel mass spectrometer (MS). After exposure (aging), prints sprinkled one of ten different formulations of hydrophobic powder. Then the prints were raised and analyzed using MALDI-TOF-MS.

Expected that will be detected is received as cholesterol, and squalene, in practice, however, was discovered only squalene. It was observed at m/z 433 corresponding to the molecular ion plus adduct of sodium. As shown in Fig.7, the intensity in the absence of any reinforcing agent is about 2,000 mW (an imprint of 12). This intensity is increased to 14,000 mW by using DHB. Among powders, those that contain the highest proportions of carbon black, give the highest response (prints 9 and 10), and the higher proportion of black carbon, the higher the intensity of the peak, while the sample with the highest intensity of about 22,000 mW is the ratio of carbon black to the diluent 1:2.

As a rule, the presence of DHB in the powder slightly increases the intensity of the peak (prints 1 and 2 titanium dioxide, and fingerprints 4 and 5 - the ratio of carbon black to the diluent 1:10), but this trend does not hold for powders that contain dyes crystal violet (crystal violet) (3 and 6) and rhodamine 6G (7 and 8), in which case get low signals, equivalent to imprint 12 without treatment or even lower signals.

Direct detection of cholesterol and squalene using MALDI-TOF-MS, using hydrophobic particles as a reinforcing matrix

Because cholesterol was not detected at MS fingerprints, tried to detect this connection using system MALDI-TOF-MS in the presence of the tvii DHB and new apadravya funds. For this purpose the standards of a mixture of squalene and cholesterol were deposited on the surface of the metal plate target and eight sets of these spots in triplicate were sprinkled eight formulations of hydrophobic powders used in 3.6, or were treated DHB, or were left without treatment.

In this case, the peaks associated with squalene, again observed at m/z 433 due to the adduct of sodium compounds and powders 1, 2, 7 and 8 give the intensity (signal) is significantly higher intensity in the presence of DHB (Fig). The highest intensity was obtained when using the red fluorescent powder (rhodamine 6G, the powder 1, 49,000 mW), however, powders with carbon black also give good responses (1:10, 6000 mV; 1:5, 36,000 mV; 1:2, 4,000 mW). The peak is now observed at m/z 386, but in all cases the magnitude of the intensities were much lower than for squalene. The highest intensity was obtained in the presence of powder, which contains the dye crystal violet (1,800 mW, powder 4), and powders with carbon black also give relatively good peaks (about 800 mV for powder 3, 1:10 plus DHB, for powder 7, 1:5, and for powder 8,1:2) (Fig.9).

The actual spectra of these two compounds is shown in figa (squalene) and fig.10b (cholesterol). Clearly the best response to squalene in the given conditions of the experiment.

Direct detection of PR is using MALDI-TOF-MS exogenous metabolites from smokers dusted latent fingerprints

As the hydrophobic powder is used the powder with the included carbon black, with the original ratio of carbon black: PTEOS equal to 1:2, the synthesis of which is described in the next section of this specification.

These particles were uniformly introduced into the mixture of fine particles of iron (diameter <60 μm) and stearic acid (1.0% by weight), so that the ratio of hydrophobic particles to particles of iron is 2:98 by weight. Latent prints were dusted with powder using serial magnetic probe and then analyzed on the location (in situ) imprints on the surface of stainless steel plates (the plates of the target Shimadzu MALDI-TOF-MS), or were raised with the use of serial lifting tape. Raised prints were attached to the plates of the target, party imprint up, using a commercial adhesive tape. Analysis of MALDI-TOF-MS was carried out using a setup Kratos Axima CFR plus MALDI-TOF-MS (Shimadzu Biotech, Manchester, UK)operating in reflection mode (reflectron positive ion. The mass matrix was used 2,5-dihydroxy benzoic acid (DHB) (10 mg/ml in a solution of 50:50 acetonitrile: deionized water [dH2O]).

The results obtained are shown figure 11, demonstrate the absence of a detected peak nicotine (162 FW) or cotinine (FM 176) in the absence of matrix reinforcing agent for latent about the signet, which was enriched with a mixture of nicotine and cotinine (10 ng each, figa). When enriched in these compounds the prints add a normal matrix a reinforcing agent in the form of a DHB, then see peaks at 163 and 177 (figs). When enriched latent prints add a hydrophobic powder, then again see peaks at 163 and 177, which suggests that this powder acts as a reinforcing agent. Observe now an additional peak at m/z 199 probably caused by the adduct of sodium cotinine. The peak at m/z 157 see in the spectrum is not enriched imprint, topped with a hydrophobic powder, together with a peak at m/z 163, having approximately the same intensity. Not see peaks at 177 and 199 in this spectrum (fig.11d). It should be borne in mind that fig.11b peak at 163 has a significantly higher intensity than the peak at 157, possibly due to the presence of nicotine.

Figure 11 clearly shows that nicotine and cotinine (both at 10 ng per imprint), with the introduction of latent fingerprints can not be detected by TOF-MS only by adding such a reinforcing matrix, as DHB (figa in comparison with figs). Figure 11 also shows that the hydrophobic powder, used for the manifestation of latent fingerprint, also acts as a reinforcing agent, capable of detecting these compounds (fig.11b). Chiefly the e peaks were found at m/z 163.3 (weight formula for nicotine 162.23) and 177.3 (weight formula for cotinine 176.22) and 199.2. This last peak is probably caused by the adduct of sodium cotinine (C10H12N2ONa, weight formula 199.21). These peaks are absent in the pre-dusted latent print that was not enriched with nicotine and cotinine (fig.11d).

The corresponding spectra for a former smoker who smoked the last 24 hours, shown in figure 2. The main peaks are not associated with a powder matrix, visible at m/z 163.1 (nicotine) and 199.1 (adduct of sodium cotinine), and we can also see a smaller peak at 177.1 (cotinine).

Interestingly, when investigating the spectra at higher mass numbers, then see peaks at 307.4 and 433.4 for prints directly deposited on a plate of stainless steel. These peaks are probably caused by the sodium adduct of stearic acid (FW 307.47) and sodium adduct of squalene (FW 433.71), respectively. It should be borne in mind that the stearic acid used in the preparation outrivaled funds, and natural squalene stands out as the endogenous component of the fingerprint.

These peaks can be used for calibration peaks obtained with a raised prints, as they are present on the surface of the lifting tape, which itself is attached to the surface of the target plate of stainless steel. This raises the surface, so that the flight time of the ionized species is now becoming the minority is e, than in the case of the steel surface, and visible m/z values of the peaks become smaller. Thus, for spectra prints from smokers on raised bands, peaks for sodium adduct visible when 305.1 and 431.2, together with peaks at 174.6 (main) and 197.4. Lower mass peaks can be adjusted to 2.3 m/z units, which gives peaks at 176.9 (cotinine) and 199.7 (adduct of sodium catenin).

Peaks at 161, 167 and 168 are permanently visible in MS only for smokers. It is known that, despite the fact that nicotine produces about 95% of the alkaloids in tobacco leaf, there are also various other alkaloids, among which the most abundant are nornicotine (RMM 148) and anatomen (RMM 160) [22]. The peak at m/z 161 may be caused by antamina as protonated forms of nicotine and cotinine observed in the spectra, however, to confirm the need for more work.

Preparation containing carbon soot agent

Main preparation method blank nanoparticles involves mixing 30 ml of ethanol, 5 ml dH2O, 2.5 ml of tetraethoxysilane (TEOS) and 2.5 ml of phenyltriethoxysilane (PTEOS) in the centrifuge. To this mixture, add 2 ml of ammonium hydroxide solution and the solution rotate throughout the night. After that make centrifugation of the suspension (3 minutes at 3,000 rpm).

The product is isolated in the following series of operations zentrifugenbau and washing with 10:90 by volume ethanol in water, then kept in suspension of 3:97 by volume ethanol in water. Then carry out the analysis of particle size distribution and SEM and TEM microscopy.

To obtain particles with carbon black, 5 ml of carbon when diluted 1:100 with water is added to a solution of the precursor. To obtain magnetic particles coated with TEOS:PTEOS, prepare powder of magnetite in accordance with the published methodology, and 5 ml of suspension in water is added to the solution predecessor.

Sources of information

1. The method of determining the presence of residue in the imprint technique of mass spectrometry using matrix, which includes the following operations:
i) coating the fingerprint powder, which can act as a matrix in the technique of mass spectrometry using matrix; and to facilitate the detection and/or imaging of the fingerprint to form the imprint of the coated particles; and
ii) carrying out mass spectrometry of the material forming the imprint of the coated particles selected from the MALDI-TOF-MS or " SALDI " -TOF-MS, and use the powder contains hydrophobic particles of silicon dioxide, and the metal, metal nitride, metal oxide or carbon, to detect the presence or absence of ostad is.

2. The method according to claim 1, wherein the metal oxide is selected from the group comprising titanium dioxide, iron oxide, hematite, and combinations thereof.

3. The method according to claim 1, wherein the carbon is selected from the group comprising carbon black, fullerene compound, carbon nanotubes, graphite and its counterpart, as well as their combinations.

4. The method according to claim 1, wherein the metal is selected from the group comprising aluminum, iron, and combinations thereof.

5. The method according to claim 1, in which particles of metal, metal nitride, metal oxide or carbon, introduced in the hydrophobic particles of silicon dioxide.

6. The method according to claim 5, in which the mean particle diameter of < 100 μm, perhaps from 10 to 90 μm, preferably from 45 to 65 μm, and preferably from 65 to 90 microns.

7. The method according to claim 5, in which the average particle diameter is ≤1 µm, possibly from 200 to 900 nm, preferably from 300 to 600 nm, and preferably from 400 to 500 nm.

8. The method according to claim 1, wherein the powder further comprises a dye molecule.

9. The method according to claim 8, in which the molecule of the dye is fluorescent or colored.

10. The method according to claim 8 or 9, in which the molecule of dye and/or painted molecule is introduced into a hydrophobic particle of silicon dioxide.

11. The method according to claim 1, in which the powder is magnetic or paramagnetic.

12. The method according to claim 1, wherein a fingerprint lifted from his seat n the bearing using lifting tape and enter into contact with the substrate sample of the mass spectrometer, then put on the fingerprint powder.

13. The method according to claim 1, in which mass spectrometry using matrix selected from the group which consists of MALDI-TOF-MS-MS or " SALDI " -TOF-MS-MS, and combinations thereof.

14. The method according to claim 1, which additionally provides for the visualization and/or imaging of a fingerprint.

15. The method according to claim 1, in which the balance represents the endogenous residue, in particular endogenous metabolite and/or exogenous metabolite.

16. The method according to item 15, in which an endogenous metabolite is a squalene.

17. The method according to item 15, in which exogenous metabolite is a metabolite of nicotine, for example catenin.

18. The method according to claim 1, in which the balance represents a contact residue.

19. The method according to p, in which the contact residue is a drug.

20. The method according to claim 19, in which a drug is a cocaine.

21. The method according to claim 1, in which the imprint together contain at least one endogenous residue and at least one contact residue.



 

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EFFECT: higher efficiency and accuracy of detection.

2 ex, 2 tbl

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