Ultra-trace analysis of illicit drugs from transfer of an electrostatic lift

This article introduces a method of collecting and analysing drug residues that integrates both electrostatic lifting and nanomanipulation-coupled to nanospray ionization mass spectrometry. The application of this hyphenated technique exhibits a useful means of collection and extraction of drug residues with ease and efficiency along with decreased limits of detection. From this method, it will be shown how increased sensitivity of analysis and lower limits of detection for drug analysis can be achieved. The same principles that allow lifting of dust prints by electrostatic lifting can be applied to lifting drug residues. Probing of the drug residues by nanomanipulation occurs directly from the lift, which provides a great platform for extraction. Nanomanipulation-coupled to nanospray ionization-mass spectrometry has been used for the extraction of trace analytes in previous experiments and is known as a very sensitive technique for the detection of ultra-trace residue. This method will demonstrate the electrostatic lifting of drug residue particles from a surface followed by extraction and ionization with nanomanipulation-nanospray ionization. The utility of this novel methodology allows for a more productive analysis when presented with ultra-trace amounts of sample.     

Application of Raman Spectroscopy and Surface‐Enhanced Raman Scattering to the Analysis of Synthetic Dyes Found in Ballpoint Pen Inks*

Abstract: The applicability of Raman spectroscopy and surface‐enhanced Raman scattering (SERS) to the analysis of synthetic dyes commonly found in ballpoint inks was investigated in a comparative study. Spectra of 10 dyes were obtained using a dispersive system (633 nm, 785 nm lasers) and a Fourier transform system (1064 nm laser) under different analytical conditions (e.g., powdered pigments, solutions, thin layer chromatography [TLC] spots). While high fluorescence background and poor spectral quality often characterized the normal Raman spectra of the dyes studied, SERS was found to be generally helpful. Additionally, dye standards and a single ballpoint ink were developed on a TLC plate following a typical ink analysis procedure. SERS spectra were successfully collected directly from the TLC plate, thus demonstrating a possible forensic application for the technique.     

Highly Sensitive Detection of Blood by Surface Enhanced Raman Scattering,,

Raman spectroscopy for forensic body fluid analysis has received some attention due to the nondestructive nature and potential application for identification at the crime scene; however, its usage has been limited by low detection sensitivity. Surface enhanced Raman scattering (SERS) was evaluated for blood identification for forensic applications. Specifically, a SERS‐active substrate was fabricated, composed of nickel nanotips coated with Ag nanoparticles. Compared with a conventional substrate, the SERS substrate enhanced Raman scattering by more than two orders of magnitude and allowed blood to be identified to a dilution of 1:100,000. Blood was also successfully detected by swabbing the SERS substrate directly on mock evidence. Most importantly, Raman spectra obtained by swabbing the SERS substrate on blood stains were free of luminescence even when blood was deposited on luminescent fabrics. The nondestructive character, simplicity of sample preparation, and high sensitivity make SERS a prime candidate for field and laboratory‐based blood identification.     

Detection of Denatonium Benzoate (Bitrex) Remnants in Noncommercial Alcoholic Beverages by Raman Spectroscopy

Illegal alcoholic beverages are often introduced into market using cheap technical alcohol, which is contaminated by denatonium benzoate (Bitrex) of very small concentration. Bitrex is the most bitter chemical compound and has to be removed before alcohol consumption. The home‐made methods utilize sodium hypochlorite to disintegrate particles of denatonium benzoate in alcohol and to remove bitter taste before trading. In this experimental studies, we propose a novel method that detects in a fast way the remnants of denatonium benzoate in dubious alcohol samples by Raman spectroscopy. This method applies a portable Raman spectrometer of excitation wavelength 785 nm and utilizes the effect of surface‐enhanced Raman spectroscopy (SERS) to recognize the suspected alcoholic beverages. High effectiveness (over 98%) of YES/NO classification of the investigated samples was observed when the nonlinear algorithm support vector machine (SVM) was exploited at carefully adjusted detection parameters. The method can identify illicit alcohol within minutes.     

Direct Analyte‐Probed Nanoextraction Coupled to Nanospray Ionization–Mass Spectrometry of Drug Residues from Latent Fingerprints

Here, we present a method of extracting drug residues from fingerprints via Direct Analyte‐Probed Nanoextraction coupled to nanospray ionization–mass spectrometry (DAPNe‐NSI‐MS). This instrumental technique provides higher selectivity and lower detection limits over current methods, greatly reducing sample preparation, and does not compromise the integrity of latent fingerprints. This coupled to Raman microscopy is an advantageous supplement for location and identification of trace particles. DAPNe uses a nanomanipulator for extraction and differing microscopies for localization of chemicals of interest. A capillary tip with solvent of choice is placed in a nanopositioner. The surface to be analyzed is placed under a microscope, and a particle of interest is located. Using a pressure injector, the solvent is injected onto the surface where it dissolves the analyte, and then extracted back into the capillary tip. The solution is then directly analyzed via NSI‐MS. Analyses of caffeine, cocaine, crystal methamphetamine, and ecstasy have been performed successfully.     

The Optimal Distance of the Electrode to the Lifting Film Surface when Lifting Dust Footwear Impressions Using an Electrostatic Dust Print Lifter

Footwear impressions are one of the valuable physical evidence encountered at crime scenes and its identification can facilitate narrowing down the suspects and establishing the identity of the criminals. The technique of electrostatic lifting (ESL) dust shoeprints at crime scenes is well established with scenes of crime examiners. And in the procedure, the recovery of the original item containing the impression should be made as good as it can and not damage the marks. In this study, the different particle sizes of SiO₂ were used to simulate dust and light soil residues, comparing different particle sizes and electrode positions were used to compare the adsorption ability on the surface of electrostatic lifting film in high‐voltage electrostatic field for the collection of shoeprints. The results indicated that lifting film in electrostatic field will be pressed down to the dust surface, the compaction and electrostatic adsorption force will vary with the distance between the high‐voltage electrode and film surface. Reducing the distance can increase the electric field strength near the lifting film, and the adsorption capacity of Mylar aluminum‐plating film to dusts can be improved significantly. Adsorption capacity of the lifting film was related to the size of dust particles and significantly improved with the increase of the particle charge. The optimum distance which can get the best adsorption capacity between the electrode and film surface is from 10 to 15 mm.     

基于银胶表面增强拉曼光谱检验蓝色书写字迹初探

目的用基于银胶的表面增强拉曼光谱技术检验蓝色签字笔字迹。方法取4种常见不同牌号的蓝色签字笔在其字迹上滴加微量银胶,用显微共焦拉曼光谱仪进行检验。结果不同品牌的蓝色签字笔字迹的表面增强拉曼光谱不同。结论基于银胶的表面增强拉曼光谱技术可用于纸张上蓝色签字笔字迹的检验。     

藻酸盐材料在提取深色织物上潜血足迹的应用

目的建立一种用藻酸盐齿科材料转印显现纺织物上血足迹的方法。方法依据藻酸盐能和纺织物上血足迹中的血结合,从而使足迹被转印到藻酸盐模上,并通过染色增强反差。结果藻酸盐印模材料转印法能够在不破坏织物客体的情况下清楚的反映出血足迹细节特征。结论藻酸盐类齿科材料能够转印、提取显现织物上血足迹,尤其是潜在血足迹,并反映出其细节特征。     

Detection of Mitragynine in Mitragyna Speciosa (Kratom) Using Surface-Enhanced Raman Spectroscopy with Handheld Devices

A simple, quick, selective, sensitive, and effective field-friendly method capable of being used by nonexperts has been developed for detecting mitragynine in Mitragyna speciosa (kratom) using surface-enhanced Raman spectroscopy (SERS). Over 100 samples and blanks (known to be either positive or negative for the presence of mitragynine) were examined in duplicate using five identical handheld Raman spectrometers, which provided a data set of over 1,000 examinations. Based on the results of these analyses, the method yielded a true-positive rate of 99.3%, a true-negative rate of 97.9%, a false-positive rate of 2.1%, and a false-negative rate of 0.7%. The average minimum detectable concentration (C_m) of mitragynine that reproducibly yielded a match for one of the library spectra on all five instruments was determined to be 342 ng/mL (ppb). This C_m value is a conservative estimate considering that the extraction process was not fully optimized by this study, which was not necessary since the C_m value achieved was well below typical mitragynine concentrations in kratom (1.3–2.3%). The method is ideal (i) for prioritizing samples for additional testing using other more time-consuming laboratory-based techniques needed to detect and quantify mitragynine and (ii) for field use at international mail facility (IMF) satellite laboratories to help interdict kratom and prevent this dangerous product from reaching the U.S. supply chain.     

The spectroscopic detection of exogenous material in fingerprints after development with powders and recovery with adhesive lifters

The application of powders to fingerprints has long been established as an effective and reliable method for developing latent fingerprints. The powders adhere to the ridge pattern of the fingerprint only, thus allowing the image to be visualised. Fingerprints developed in situ at a crime scene routinely undergo lifting with specialist tapes to facilitate subsequent laboratory analysis. As with all recovered evidence these samples would be stored in evidence bags to allow secure transit from the scene to the laboratory and also to preserve the chain of evidence. In this paper, the application of Raman spectroscopy for the analysis of exogenous material in latent fingerprints is reported for contaminated fingerprints that had been treated with powders and also subsequently lifted with adhesive tapes. A selection of over the counter (OTC) analgesics were used as samples for the analysis and contaminated fingerprints were deposited on clean glass slides. The application of aluminium or iron based powders to contaminated fingerprints did not interfere with the Raman spectra obtained for the contaminants. In most cases background fluorescence attributed to the sebaceous content of the latent fingerprint was reduced by the application of the powder thus reducing spectral interference. Contaminated fingerprints developed with powders and then lifted with lifting tapes were also examined. The combination of these two techniques did not interfere with the successful analysis of exogenous contaminants by Raman spectroscopy. The lifting process was repeated using hinge lifters. As the hinge lifters exhibited strong Raman bands the spectroscopic analysis was more complex and an increase in the number of exposures to the detector allowed for improved clarification. Raman spectra of developed and lifted fingerprints recorded through evidence bags were obtained and it was found that the detection process was not compromised in any way. Although the application of powders did not interfere with the detection process the time taken to locate the contaminant was increased due to the physical presence of more material within the fingerprint. The presence of interfering Raman bands from lifting tapes is another potential complication. This, however, could be removed by spectral subtraction or by the choice of lifting tapes that have only weak Raman bands.     

Chemical enhancement of footwear impressions in blood deposited on fabric — Evaluating the use of alginate casting materials followed by chemical enhancement

Most footwear marks made in blood on a surface such as fabric tend to be enhanced in situ rather than physically recovered using a lifting technique prior to enhancement. This work reports on the use of an alginate material to recover the impressed footwear marks made in blood and deposited on a range of fabric types and colours. The lifted marks were then enhanced using acid black 1 and leuco crystal violet with excellent results.This presents a new method for the lifting and recovery of blood impressions in situ from crime scene followed by subsequent mark enhancement of the lifted impression.     

Elucidation of the effect of heat exposure on hair colored by permanent and semipermanent colorants using surface-enhanced Raman spectroscopy

Confirmatory identification of hair colorants can be used to establish a connection between a suspect and the crime science or demonstrate the absence of such connections. A growing body of evidence shows that surface-enhanced Raman spectroscopy (SERS) could be a confirmatory, minimally destructive, and fully noninvasive analysis of hair colorants. In SERS, a signal that provide the information about the chemical structure of both permanent and semipermanent dyes present on hair is enhanced by a million-fold using noble metal nanostructures. However, it is unclear whether the information of hair colorants can be revealed if hair was contaminated or exposed to harsh environments such as sunlight and heat. In this work, we determine the effect of a short- and long-term heat exposure on SERS-based analysis of hair colored with blue and red permanent and semipermanent dyes. We found that short and especially long-term heat exposure at 220°C could alter chemical structure, and consequently SERS spectra, of permanent and semipermanent colorants. This thermal degradation of permanent dyes complicates their direct identification using SERS. We also found that partial least squares discriminant analysis can be used to overcome this issue allowing for highly accurate identification of both permanent and semipermanent dyes on colored hair that was exposed to 220°C for 6–12 min. These results show that heat exposure of colored hair should be strongly considered upon their SERS-based examination to avoid both false positive or false negative identification of chemical dyes.     

A comparative research of two lifting methods: electrostatic lifter and gelatin lifter

Two-dimensional dust shoeprints are often of very high resolution and contain unique features. Lifting these prints in the most effective method may contribute much to preserving these fine details. A research was conducted by experts from Israel and Switzerland to compare gelatin lifters and electrostatic lifters for lifting shoeprints. Several substrates were chosen, and on each material a set of dry dust shoeprints was made. A set of wet prints was made on paper as well. The shoeprints were approximately of the same quality, and the only variable was the nature of the material. On substrates indifferent to the method used, the preferable sequence was tested. Gelatin lifter was superior on most substrates and for wet prints. The superior sequence for using both methods is electrostatic lifting followed by gelatin lifter.     

Evaluation and classification of fentanyl-related compounds using EC-SERS and machine learning

Multiple analytical techniques for the screening of fentanyl-related compounds exist. High discriminatory methods such as GC–MS and LC–MS are expensive, time-consuming, and less amenable to onsite analysis. Raman spectroscopy provides a rapid, inexpensive alternative. Raman variants such as electrochemical surface-enhanced Raman scattering (EC-SERS) can provide signal enhancements with 10¹⁰ magnitudes, allowing for the detection of low-concentration analytes, otherwise undetected using conventional Raman. Library search algorithms embedded in instruments utilizing SERS may suffer from accuracy when multicomponent mixtures involving fentanyl derivatives are analyzed. The complexing of machine learning techniques to Raman spectra demonstrates an improvement in the discrimination of drugs even when present in multicomponent mixtures of various ratios. Additionally, these algorithms are capable of identifying spectral features difficult to detect by manual comparisons. Therefore, the goal of this study was to evaluate fentanyl-related compounds and other drugs of abuse using EC-SERS and to process the acquired data using machine learning—convolutional neural networks (CNN). The CNN was created using Keras v 2.4.0 with Tensorflow v 2.9.1 backend. In-house binary mixtures and authentic adjudicated case samples were used to evaluate the created machine-learning models. The overall accuracy of the model was 98.4 ± 0.1% after 10-fold cross-validation. The correct identification for the in-house binary mixtures was 92%, while the authentic case samples were 85%. The high accuracies achieved in this study demonstrate the advantage of using machine learning to process spectral data when screening seized drug materials comprised of multiple components.     

Screening tablets for DOB using surface-enhanced Raman spectroscopy

2,5,-Dimethoxy-4-bromoamphetamine (DOB) is of particular interest among the various "ecstasy" variants because there is an unusually long delay between consumption and effect, which dramatically increases the danger of accidental overdose in users. Screening for DOB in tablets is problematic because it is pharmacologically active at 0.2-3 mg, which is c. 50 times less than 3,4-methylenedioxy-N-methylamphetamine (MDMA) and makes it more difficult to detect in seized tablets using conventional spot tests. The normal Raman spectra of seized DOB tablets are dominated by the bands of the excipient with no evidence of the drug component. Here we report the first use of on-tablet surface-enhanced Raman spectroscopy (SERS) to enhance the signal from a low concentration drug. Raman studies (785-nm excitation) were carried on series of model DOB/lactose tablets (total mass c. 400 mg) containing between 1 mg and 15 microg of DOB and on seized DOB tablets. To generate surface-enhanced spectra, 5 microL of centrifuged silver colloid was dispensed onto the upper surface of the tablets, followed by 5 microL of 1.0 mol/dm(3) NaCl. The probe laser was directed onto the treated area and spectra accumulated for c. 20 sec (10 sec x 2). It was found that the enhancement of the DOB component in the model tablets containing 1 mg DOB/tablet and in the seized tablets tested was so large that their spectra were completely dominated by the vibrational bands of DOB with little or no contribution from the unenhanced lactose excipient. Indeed, the most intense DOB band was visible even in tablets containing just 15 microg of the drug. On-tablet surface-enhanced Raman spectroscopy is a simple method to distinguish between low dose DOB tablets and those with no active constituent. The fact that unique spectra are obtained allows identification of the drug while the lack of sample preparation and short signal accumulation times mean that the entire test can be carried out in <1 min.     

A preliminary investigation into the use of alginates for the lifting and enhancement of fingermarks in blood

Recent studies have reported the use of alginate in the lifting and subsequent enhancement of footwear marks in blood. A study was set up to assess the use of such a method in the treatment of fingermarks in blood on a variety of porous, non-porous and semi-porous surfaces. Other variables included ageing of the fingermarks in blood and the application of chemicals prior to or post-alginate lifting. All different variations were compared to direct chemical treatment of the substrate. The results demonstrated that alginate is not compatible with certain substrates (e.g. glass and tile). On substrates that were compatible with alginate (e.g. fabric and paper), the enhanced fingermarks on the alginate cast and the enhanced fingermarks on the post-alginate substrates appeared, overall, inferior compared to direct chemical enhancement without the use of alginate. A further variation using water-based protein stains directly mixed with the alginate appeared to provide enhancement directly on the substrate as well as simultaneous lifting and enhancing the fingermarks in blood on the alginate cast.     

Discrimination of phenethylamine regioisomers and structural analogues by Raman spectroscopy

In this study, the Raman spectra of 21 phenethylamines were obtained using far‐red excitation (785 nm). The distinguishing ability of Raman for phenethylamines, especially for phenethylamine regioisomers and structural analogues, was investigated. Here, the evaluation of a cross section of Raman spectra demonstrated that all types of phenethylamines were distinguishable, even for certain structural analogues with high spectrum similarity. Raman exhibited high distinguishing ability for phenethylamine regioisomers that differ in the substitution position of halogen, methoxy, alkyl, or other substituted groups; as well as for structural analogues containing different groups, such as furanyl, 2,3‐dihydrofuranyl, halogen, and alkyl substituted at the same position. The Raman spectra for homologues with differences in only a methyl group were found to be highly similar; however, their spectra demonstrated small but detectable differences. Four analogue mixtures and 59 seized samples were also analyzed to study the practical use of the Raman method in forensic field. 95% of the seized samples were correctly identified, which significantly validated the ability of Raman method in identifying the correct isomers. Accordingly, this study provides a non‐destructive, high‐throughput and minimal sample preparation technique for the discrimination of phenethylamines.     

In search of blood--detection of minute particles using spectroscopic methods

An examination protocol for rapid detection of remnants of blood particles on garments of suspects in bloody murder cases is described. Microparticles of blood are sampled along with fibres and hairs using the tape lifting method. The tapings are searched with a low power microscope for red particles with morphology similar to blood. Presumed blood traces are further examined using microspectrophotometry on the cut out piece of taping. The typical visible spectrum of haemoglobin is characteristic for blood. Alternatively Raman spectroscopy can be used to measure the characteristic vibrational spectrum of haemoglobin. At a later stage, these particles may be removed from the piece of taping in order to extract the blood and attempt to obtain a genetic profile.     

DNA Profiles from Fingerprint Lifts—Enhancing the Evidential Value of Fingermarks Through Successful DNA Typing

This study evaluated the compatibility of the most common enhancement methods and lifting techniques with DNA profiling. Emphasis is placed on modern lifting techniques (i.e., gelatin lifters and Isomark?) and historical fingerprint lifts for which limited research has been previously conducted. A total of 180 fingerprints were deposited on a glass surface, enhanced, lifted, and processed for DNA typing. DNA could be extracted and profiled for all the powders and lifts tested and from both groomed fingerprints and natural prints with no significant difference in the percentage of profile recovered. DNA profiles could also be obtained from historical fingerprint lifts (79.2% of 72 lifts) with one or more alleles detected. These results demonstrate the compatibility between different powder/lift combinations and DNA profiling therefore augmenting the evidential value of fingerprints in forensic casework.