Quantitative vapor delivery for improved canine threshold testing.

The canine olfactory system is a highly efficient and intricate tool often exploited by humans for detection for its many attributes, including impressive sensitivity to trace analyte vapors. Canine detectors are often touted as having lower limits of detection, or olfactory detection threshold (ODT), than other field-relevant detection technologies; however, previous attempts to quantify canine ODTs have resulted in reported estimates spanning multiple orders of magnitude, even for the same analyte. A major contributor to these discrepancies is the vapor delivery method used for testing, where losses due to adsorption and dilution are often unaccounted for, and the presence of unattended compounds in the vapor stream due to carryover may go unnoticed. In this research, a trace vapor generator (TV-Gen) was used to deliver quantitatively accurate amounts of vapor reproducibly over time for canine testing. Analyte losses due to adsorption to surfaces in the flow path, dilution in the sniff port at the outlet, and analyte carryover were considered. Computational fluid dynamic (CFD) modeling was used to visualize analyte vapor spread throughout the port. CFD simulations revealed the need for a diffuser to encourage the diffusion of the analyte throughout the port. As a result, the modified vapor generator provides analyte air as a diffuse flow that is evenly distributed through the custom sampling orifice, as opposed to a narrow stream of air at the chosen concentration which exits directly into the environment. Laboratory validations were carried out for three analytes, amyl acetate, 2,4-dinitrotoluene (DNT), and methyl benzoate. A linear response across more than two orders of magnitude vapor concentration range was achieved for all analytes. These efforts will be applied in further research utilizing this TV-Gen vapor delivery system for canine ODT testing, eliminating many quantitative changes seen previously. Graphical abstract.

Comparative analysis of vapor profiles of fentalogs and illicit fentanyl.

Availability of fentanyl is at a record high with 3138 kg of fentanyl and related substances being seized in 2019. Fentanyl's high toxicity makes a lethal dose for most mere milligrams. With such a high potency and a consistent rise of abuse, the chances of injury or death of frontline workers increase with every interaction. Development of a non-contact detection method for fentanyl would decrease the chances of a workplace mishap. To aid in the development of a non-contact detection method, target analytes in the vapor profile of fentanyl need to be identified. In order to achieve this goal, semi-quantitative headspace analysis of fentanyl analogs and confiscated fentanyl exhibits was accomplished using solid-phase microextraction and gas chromatography coupled with mass spectrometry (SPME-GC-MS). The vapor signatures of these samples were compared to a previously reported reference-grade fentanyl vapor signature to determine the target analyte(s) for fentanyl detection in the vapor phase. A total of 20 fentalogs and confiscated exhibits, with masses ranging from 2 to 19 mg, were sampled. N-Phenylpropanamide(NPPA) or N-phenethyl-4-piperidone(NPP) was identified as target analytes in 75% of these samples. This is a crucial component for the development of a non-contact detection method for fentanyl.

A Method for Controlled Odor Delivery in Olfactory Field-Testing.

A widely recognized limitation in mammalian olfactory research is the lack of current methods for measuring odor availability (i.e., the quantifiable amount of odor presented and thus available for olfaction) of training or testing materials during behavioral or operational testing. This research utilized an existing technology known as Controlled Odor Mimic Permeation Systems (COMPS) to produce a reproducible, field-appropriate odor delivery method that can be analytically validated and quantified, akin to laboratory-based research methods, such as permeation devices that deliver a stable concentration of a specific chemical vapor for instrumental testing purposes. COMPS were created for 12 compounds across a range of carbon chain lengths and functional groups in such a way to produce similar permeation rates for all compounds. Using detection canines as a model, field-testing was performed to assess the efficacy of the method. Additionally headspace concentrations over time were measured as confirmation of odor availability using either externally sampled internal standard-solid phase microextraction-gas chromatography-mass spectrometry (ESIS-SPME-GC-MS) or collection onto a programmable temperature vaporizing (PTV) GC inlet with MS detection. Finally, lifetime usage was considered. An efficient method for producing and measuring reliable odor availabilities across various chemical functional groups was developed, addressing a noted gap in existing literature that will advance canine and other nonhuman mammal research testing.

Physical properties of odorants affect behavior of trained detection dogs during close-quarters searches.

Trained detection dogs have a unique ability to find the sources of target odors in complex fluid environments. How dogs derive information about the source of an odor from an odor plume comprised of odorants with different physical properties, such as diffusivity, is currently unknown. Two volatile chemicals associated with explosive detection, ammonia (NH3, derived from ammonium nitrate-based explosives) and 2-ethyl-1-hexanol (2E1H, associated with composition C4 plastic explosives) were used to ascertain the effects of the physical properties of odorants on the search behavior and motion of trained dogs. NH3 has a diffusivity 3.6 times that of 2E1H. Fourteen civilian detection dogs were recruited to train on each target odorant using controlled odor mimic permeation systems as training aids over 6 weeks and then tested in a controlled-environment search trial where behavior, motion, and search success were analyzed. Our results indicate the target-odorant influences search motion and time spent in the stages of searching, with dogs spending more time in larger areas while localizing NH3. This aligns with the greater diffusivity of NH3 driving diffusion-dominated odor transport when dogs are close to the odor source in contrast to the advection-driven transport of 2E1H at the same distances.

Calibrating canines-a universal detector calibrant for detection dogs.

Since the advent of the Universal Detector Calibrant (UDC) by scientists at Florida International University in 2013, this tool has gone largely unrecognized and under-utilized by canine scent detection practitioners. The UDC is a chemical that enables reliability testing of biological and instrumental detectors. Training a biological detector, such as a scent detection canine, to respond to a safe, non-target, and uncommon compound has significant advantages. For example, if used prior to a search, the UDC provides the handler with the ability to confirm the detection dog is ready to work without placing target odor on site (i.e., a positive control), thereby increasing handler confidence in their canine and providing documentation of credibility that can withstand legal scrutiny. This review describes the UDC, summarizes its role in canine detection science, and addresses applications for UDC within scent detection canine development, training, and testing.

Evaluation of canine training aids containment for homemade explosive and components by headspace analysis and canine testing.

While canines are most commonly trained to detect traditional explosives, such as nitroaromatics and smokeless powders, homemade explosives (HMEs), such as fuel-oxidizer mixtures, are arguably a greater threat. As such, it is imperative that canines are sufficiently trained in the detection of such HMEs. The training aid delivery device (TADD) is a primary containment device that has been used to house HMEs and HME components for canine detection training purposes. This research assesses the odor release from HME components, ammonium nitrate (AN), urea nitrate (UN), and potassium chlorate (PC), housed in TADDs. Canine odor recognition tests (ORTs) were used with analytical data to determine the detectability of TADDs containing AN, UN, or PC. Headspace analysis by gas chromatography/mass spectrometry (GC/MS) with solid-phase microextraction (SPME) or online cryotrapping were used to measure ammonia or chlorine, as well as other unwanted odorants, emanating from bulk AN, UN, and PC in TADDs over 28 weeks. The analytical data showed variation in the amount of ammonia and chlorine over time, with ammonia from AN and UN decreasing slowly over time and the abundance of chlorine from PC TADDs dependent on the frequency of exposure to ambient air. Even with these variations in odor abundance, canines previously trained to detect bulk explosive HME components were able to detect all three targets in glass and plastic TADDs for at least 18 months after loading. Detection proficiency ranged from 64% to 100% and was not found to be dependent on either age of material.

Non-contact detection of fentanyl by a field-portable ion mobility spectrometer.

Rapid on-site detection of fentanyl is paramount for the safety of law enforcement and other first responders. Due to the opioid epidemic, death by overdose is at an all-time high with fentanyl adulteration as the main assailant. Providing a user-friendly method for the presumptive detection of fentanyl will increase safety for first responders. Ion mobility spectrometry (IMS) provides a quick, affordable, and accurate method for detecting fentanyl. Currently, most methods for detecting fentanyl require manipulation or handling of the highly potent substance. A recent comparative analysis study on the headspace of fentanyl determined N-phenylpropanamide (NPPA) a target analyte for fentanyl enabling vapor detection. Here, we demonstrate the development of a handheld IMS method for vapor detection of the target analyte for fentanyl. An alarm was programmed into the handheld IMS device for the detection of NPPA. The system was able to accurately detect NPPA in samples of reference-grade fentanyl and diluted reference-grade fentanyl, as well as 3.67 mg of fentanyl from samples confiscated from the US border. Common adulterants and over-the-counter drugs were tested and resulted in a false alarm rate of 0 for substances sampled. The limit of detection was determined to be as low as 5 ng of NPPA. Overall, the development of this user-friendly, non-contact method has considerable promise for near real-time non-contact detection of fentanyl increasing safety of first responders.

Collection and identification of human remains volatiles by non-contact, dynamic airflow sampling and SPME-GC/MS using various sorbent materials.

Human remains detection canines are used in locating deceased humans in diverse scenarios and environments based on odor produced during the decay process of the human body. It has been established that human remains detection canines are capable of locating human remains specifically, as opposed to living humans or animal remains, thus suggesting a difference in odor between the different sources. This work explores the collection and determination of such odors using a dynamic headspace concentration device. The airflow rate and three sorbent materials-Dukal cotton gauze, Johnson & Johnson cotton-blend gauze, and polyester material-used for odor collection were evaluated using standard compounds. It was determined that higher airflow rates and openly woven material, e.g., Dukal cotton gauze, yielded significantly less total volatile compounds due to compound breakthrough through the sorbent material. Collection from polymer- and cellulose-based materials demonstrated that the molecular backbone of the material is a factor in compound collection as well. Volatiles, including cyclic and straight-chain hydrocarbons, organic acids, sulfides, aldehydes, ketones, and alcohols, were collected from a population of 27 deceased bodies from two collection locations. The common compounds between the subjects were compared and the odor profiles were determined. These odor profiles were compared with those of animal remains and living human subjects collected in the same manner. Principal component analysis showed that the odor profiles of the three sample types were distinct.

Canine olfactory detection of trained explosive and narcotic odors in mixtures using a Mixed Odor Delivery Device.

Like using a substandard calibrant to test and calibrate an instrumental detector, when detection canines are regularly exposed to less than optimal training material, their detection proficiency is diminished, risking the lives of their handlers and civilians they are intended to protect. This research examined canine detection proficiency to odor mixtures and the use of mixture training to improve said proficiency. Trained detection canines were tested on their ability to correctly locate their trained target odors, explosives or narcotics, in various mixtures from a series of blanks and distractor odors. After making base measurements, canines were trained on the target odor in mixtures using the Mixed Odor Delivery Device (MODD), which was previously developed to safely contain separated explosive components and deliver the mixed odor to a canine detector for training purposes. Headspace measurements, made using solid phase microextraction with gas chromatography/mass spectrometry (SPME-GC/MS), were also taken of mixture components in and out of the MODD to confirm that odor mixtures were accurately portrayed to the canines during MODD training. Following mixture training, canines were retested on the same mixtures. Results of the headspace analysis showed that the MODD did not alter the delivery of the odorants from the mixture components. As such, canines showed an improved proficiency in detection of target mixtures following mixture training, increasing the detection rate from 63% to 72% for pseudo cocaine mixtures and from 19% to 100% for explosive mixtures.

Controlled Odor Mimic Permeation Systems for Olfactory Training and Field Testing.

The Controlled Odor Mimic Permeation System (COMPS) was developed to provide a convenient field testing method of odor delivery at controlled and reproducible rates. COMPS are composed of an odorant of interest on an absorbent material sealed inside of a permeable polymer bag. The permeable layer allows for a constant release of the odorant over a given amount of time. The permeable bag is further stored in a secondary, impermeable bag. The double-containment procedure allows for equilibration of the odorant from the permeable bag but within the impermeable outer layer, resulting in an instant and reproducible source of odorant vapor upon removal from the outer packaging. COMPS are used in both olfactory testing for experimental scenarios and for olfactory detection training, such as with detection canines. COMPS can be used to contain a wide range of odorants (e.g., narcotics powders) and provide a controlled release of the associated odorants. Odor availability from COMPS is expressed in terms of permeation rate (i.e., the rate of the odorant vapor released from a COMPS per unit time) and is typically measured by gravimetric means. The permeation rate for a given mass or volume of odorant can be adjusted as needed by varying the bag thickness, surface area, and/or polymer type. The available odor concentration from a COMPS can also be measured by headspace analysis techniques such as solid phase microextraction with gas chromatography/mass spectrometry (SPME-GC/MS).

Trace vapor generator for Explosives and Narcotics (TV-Gen).

The Trace Vapor Generator for Explosives and Narcotics (TV-Gen) is a portable and compact instrument designed to deliver a continuous source of trace-level vapors and vapor mixtures. It provides a tool to assist in the independent validation and verification of new materials and sensors under development for the vapor detection of explosives and narcotics. The design was conceived for use with a broad range of analytes, detection systems, materials, and sensors and to switch easily between the clean and analyte vapor streams. The TV-Gen system utilizes nebulization of aqueous analyte solutions, an oven to promote efficient transport, and a control box that provides dedicated computer control with logging capabilities. Resultant vapor streams are stable over several hours, with the vapor concentration controlled by a combination of aqueous analyte solution concentration, liquid flow rate through the nebulizer, and volume flow rate of air through the TV-Gen manifold.

Methodological Considerations in Canine Olfactory Detection Research.

Dogs are increasingly used in a wide range of detection tasks including explosives, narcotics, medical, and wildlife detection. Research on detection dog performance is important to understand olfactory capabilities, behavioral characteristics, improve training, expand deployment practices, and advance applied canine technologies. As such, it is important to understand the influence of specific variables on the quantification of detection dog performance such as test design, experimental controls, odor characteristics, and statistical analysis. Methods for testing canine scent detection vary influencing the outcome metrics of performance and the validity of results. Operators, management teams, policy makers, and law enforcement rely on scientific data to make decisions, design policies, and advance canine technologies. A lack of scientific information and standardized protocols in the detector dog industry adds difficulty and inaccuracies when making informed decisions about capability, vulnerability, and risk analysis. Therefore, the aim of this review is to highlight important methodological issues and expand on considerations for conducting scientifically valid detection dog research.

Generalization and Discrimination of Molecularly Similar Odorants in Detection Canines and the Influence of Training.

Operationally-deployed canine detectors are often trained on one or a limited number of materials representing a single target odor, and training frequently occurs using materials of a high purity grade in controlled scenarios with minimal other background odors. Conversely, in the field, canine detectors are expected to generalize and identify variations of the target odor, while discriminating from similar extraneous or background odors. This exemplifies the balance between generalization and discrimination required for effective canine detectors. This research explored the tendency for detection canines to generalize or discriminate between similar odorants. Two groups of related odorants were used in two separate studies; (1) odorants of similar functional groups with differing carbon chains, and (2) odorants of similar carbon chain length but differing functional groups. Within each odorant set, the effect of training was addressed by incrementally increasing the number of odorants each canine was trained to detect. Initially, discrimination increased with increasing molecular dissimilarity in both odorant groups. After further training on additional related odorants, generalization increased across the set of odorants of the same carbon chain length, but there were no significant changes in either generalization or discrimination across the set of odorants of the same functional group. The results suggest that the canines in this study were more likely to generalize across compounds of the same chain length with differing functional group than across compounds of the same functional group, but differing chain lengths. Furthermore, some variation in performance between individual canines indicated that the tendency to generalize differed with experience, breed, and other factors affecting olfaction.

Headspace analysis of potassium chlorate using on-fiber SPME derivatization coupled with GC/MS.

Potassium chlorate is a powerful oxidizer salt that, when mixed with a fuel source, has been used as a homemade explosive (HME). As an inorganic salt, potassium chlorate has no appreciable vapor pressure under ambient conditions and requires temperatures exceeding 300 °C for decomposition. However, detection of potassium chlorate by trained canines has been demonstrated, implying that it exudes a vapor signature with one or more volatile compounds, although no such species have been confirmed to date. In this work, solid-phase microextraction with a novel on-fiber derivatization reaction was used to interrogate the headspace of several potassium chlorate samples of varying purity, as well as that of related chlorinated salts and explosive mixtures. This analysis showed the presence of few volatile species in the headspace of potassium chlorate other than vaporous chlorine, detected as the derivatized product, chloro-2-propanol. Relative amounts of chloro-2-propanol could be compared between potassium chlorate variants, and could be detected in the presence of other volatile species associated with the fuels.

Quantitative analysis of vaporous ammonia by online derivatization with gas chromatography - mass spectrometry with applications to ammonium nitrate-based explosives.

A novel method for the detection of vaporous products was developed utilizing a derivatizing agent collected onto a cryo-cooled gas chromatograph (GC) inlet, with analysis by gas chromatography-mass spectrometry (GC-MS). The technique was applied to the detection of ammonia, which has been difficult to detect at trace levels, particularly in the presence of other chemical interferents, due to its small mass and high volatility. To address this, the ammonia is derivatized in the inlet with butyl chloroformate to produce butyl carbamate, a compound that is retained by GC columns and compatible with standard GC-MS analysis. This method was then used to quantify the ammonia headspace vapor concentration produced from the dissociation of bulk ammonium nitrate as well as from mixtures with aluminum and petroleum jelly, which are fuels commonly used in homemade explosives (HMEs).

Effect of re-use of surface sampling traps on surface structure and collection efficency for trace explosive residues.

In security settings, explosive residues or particles are collected by swiping the object of interest (e.g., luggage or package) with a collection medium, or trap. Particles on the trap are thermally desorbed for detection by ion mobility spectrometry (IMS) or other analyses. A high trap sampling efficiency increases the chance of detection, and is affected by a number of factors. In particular, this work studies the effect of trap re-use on collection efficiency of organic explosives, namely 2,4,6-trinitrotoluene (TNT) and 1,3,5-trinitro-1,3,5-triazinane (RDX), and correlates this data to quantifiable morphology changes. Collection efficiency was measured by liquid extraction of the traps with detection and quantitation by gas chromatography / mass spectrometry (GC/MS). Using silhouette microscopy for visualization of the trap texture, morphology changes were quantified by several measurements of trap roughness and hairiness, drawing from techniques and metrics used in the textiles industry. Nomex traps were visibly roughened by repeated re-use, and this was correlated with significant improvements in trap collection efficiency (11-57%) depending upon the specific analyte and substrate combination interrogated. Teflon-coated fiberglass (TCFG) traps showed little change with repeated swiping and minimal to no improvement in particle collection efficiency. These results have direct implications for optimizing particle collection traps for use in security settings.

Mixed Vapor Generation Device for delivery of homemade explosives vapor plumes.

While there is a large body of research on the properties and detection of traditional military high explosives and propellant low explosives, there is a dearth of research on homemade explosive (HME) materials, though they are prevalent today. The safety of working with these materials in the laboratory is the greatest limiting factor preventing HME research. A vapor delivery tool, the Mixed Vapor Generation Device (MV-Gen), was designed to safely contain the individual solid or liquid components that often compose homemade explosives vapor plumes and deliver the mixed component vapors for instrumental sampling and analysis. Within the MV-Gen, each component is separated and only the vapors mix as they are carried through the device by flowing air. The resulting mixed vapor is representative of either mixed explosive material or bulk explosives. Component materials are held in up to four individual, removable vials with vapor concentrations controlled by vial orifice size, temperature, and diluent airflow. The total concentration can be adjusted by altering vial temperature via a thermal water jacket surrounding the entirety of the device, or by adjusting the flow rate of diluent air through the device. The MV-Gen was evaluated first with surrogate compounds, followed by two types of homemade explosives, to include a binary explosive mixture and a peroxide explosive. To evaluate the device, vapors were cold-trapped on an online sampling system and analyzed by gas chromatography/mass spectrometry. It was determined that the device yielded reproducible vapor concentrations of both single and mixed components, and the ratio of these vapors can be easily adjusted to mimic varying forms of homemade explosives.

Feasibility of Canine Detection of Mass Storage Devices: A Study of Volatile Organic Compounds Emanating from Electronic Devices Using Solid Phase Microextraction.

Detection of canines are well-known to be valuable in the location of contraband, such as explosives or narcotics. More recently, canines have been trained and utilized in the detection of concealed mass storage devices that might contain evidence of illegal activity such as child pornography. To lay the analytical foundation for this detection work, research was carried out to determine the volatile organic compounds associated with mass storage devices (MSD) that could be used by trained canines for detection. Headspace analysis of a variety of electronic devices was performed using solid phase microextraction (SPME) with gas chromatography/mass spectrometry (GC/MS). Analyses found several volatile compounds common to SIM and SD cards, as well as USB drives, including 2-propenenitrile, styrene, isophorone, hydroxycyclohexyl phenyl ketone, and 2-furanmethanol, tetrahydro. Results indicated that mass storage devices do have a characteristic odor profile making detection with minimal false alerts feasible for trained canines.

Factors affecting the intramolecular decomposition of hexamethylene triperoxide diamine and implications for detection.

Hexamethylene triperoxide diamine (HMTD) is an easily synthesized and highly sensitive organic peroxide frequently used as a primary explosive. The vapor pressure of HMTD is very low, impeding vapor detection, especially when compared to other peroxide explosives, such as triacetone triperoxide (TATP) or diacetone diperoxide (DADP). Despite this fact, HMTD has a perceptible odor that could be utilized in the indirect detection of HMTD vapor. Headspace measurements above solid HMTD samples confirm that HMTD readily decomposes under ambient conditions to form highly volatile products that include formic acid, ammonia, trimethylamine and formamides. The presence and quantity of these compounds are affected by storage condition, time, and synthetic method, with synthetic method having the most significant effect on the content of the headspace. A kinetic study of HMTD decomposition in solution indicated a correlation between degradation rate and the presence of decomposition species identified in the headspace, and provided further insight into the mechanism of decomposition. The study provided evidence for a proton assisted decomposition reaction with water, as well as an intramolecular decomposition process facilitated by the presence of water.

Technical note: Headspace analysis of explosive compounds using a novel sampling chamber.

The development of instruments and methods for explosive vapor detection is a continually evolving field of interest. A thorough understanding of the characteristic vapor signatures of explosive material is imperative for the development and testing of new and current detectors. In this research a headspace sampling chamber was designed to contain explosive materials for the controlled, reproducible sampling and characterization of vapors associated with these materials. In a detonation test, the chamber was shown to contain an explosion equivalent to three grams of trinitrotoluene (TNT) without damage to the chamber. The efficacy of the chamber in controlled headspace sampling was evaluated in laboratory tests with bulk explosive materials. Small quantities of TNT, triacetone triperoxide (TATP) and hexamethylene triperoxide diamine (HMTD) were separately placed in the sampling chamber, and the headspace of each material was analyzed by gas chromatography/mass spectrometry (GC/MS) with online cryogenic trapping to yield characteristic vapor signatures for each explosive compound. Chamber sampling conditions, temperature and sampling time, were varied to demonstrate suitability for precise headspace analysis.