September 23, 2019 by Locard's Lab

Rapid & Safer Drug Testing with Mass Spectrometry

Scientists have demonstrated a new method that allows police to quickly predict the contents of suspected drugs packages using Direct Analysis in Real Time mass spectrometry.

When faced with a suspected illicit drug, police officers will not know the identity or potency of the substance. Investigators will often perform something called a presumptive test to indicate what kind of drug they are dealing with. These tests typically involve scooping up a small amount of the material and adding a few drops of a specific chemical reagent to the sample. If the drug is present, a chemical reaction will occur resulting in a distinctive colour change to indicate a positive result.

Unfortunately, these tests can be somewhat hazardous, necessitating both the potential exposure to harmful substances in order to sample the drug, as well as the use of chemicals in the field. This has become particularly problematic in recent years as more uncharacterised and potentially harmful illicit drugs have hit the market. For instance, recent news reports have seen first responders complain of exposure to the drug fentanyl, a particularly potent opioid harmful even in small quantities.

In light of this, the need for rapid, reliable and safe drug screening techniques is greater than ever before.

Researchers at the National Institute of Standards and Technology (NIST) have teamed up with Maryland State Police and Vermont Forensic Laboratory to develop a method of quickly predicting the contents of a suspicious package without the need to handle the contents. We’ve all seen how airport staff check luggage for traces of explosives. They swipe an absorbent material over the surface of the bag, introduce that swab to an analyser, and receive a rapid alert to the presence of specific controlled substances. This new method essentially follows the same process. Upon discovering a package suspected of containing illicit drugs, the investigator simply swipes an absorbent wipe across the surface of the package, and that swab is then exposed to the mass spectrometer.

The new method utilises Direct Analysis in Real Time mass spectrometry (DART-MS) analysis. DART-MS is a form of ambient ionisation mass spectrometry in which the sample is placed between the DART ion source and the inlet of the mass spectrometer, allowing chemical components in the sample to be ionised and drawn into the mass spectrometer for rapid analysis. The major advantage of this technique is that it requires no form of sample preparation, so the sample itself (or in this case a swab of the sample) can be analysed directly for almost instantaneous results. You can read more about how DART works here.

In a study recently published in the journal Forensic Science International, the technique was applied to almost 200 suspicious packages, including plastic baggies, pill bottles, envelopes, and tin foil bags. From these samples, a range of illicit substances were detected, including fentanyl, cocaine, heroin, methamphetamine and synthetic cannabinoids. Also detected on a couple of the packages was carfentanil, a particularly powerful opioid thousands of times more potent than heroin and a great concern for first responders. It was shown that only a few micrograms of drug on the outside of the packages could be detected. The study has successfully demonstrated the possibility of rapidly screening for the presence of harmful drugs such as fentanyl, even when mixed with other substances like heroin. In fact, the contents of the packages were successfully predicted 92% of the time.

This new technique enables police officers to rapidly screen suspicious bags and packages for the presence of illicit drugs, whilst reducing the risk of exposure to harmful substances. By quickly and safely predicting the contents of a package, steps can then be taken to ensure the evidence is processed under appropriate conditions, for instance in a laboratory fume hood and with appropriate protective equipment. Although a complete analysis by standardised laboratory methods would still be required for legal purposes, this new technique at least allows for the rapid, and safe screening of suspected drugs.

Sisco, E. Robinson, E. L. Burns, A. Mead, R. What’s in the Bag? Analysis of Exterior Drug Packaging by TD-DART-MS to Predict the Contents. For. Sci. Int., 2019, In Press.

May 12, 2019 by Locard's Lab

Detecting Homemade Bombs & Explosives in Sweat

Improvised explosive devices (IEDs) are often used in the implementation of terrorist attacks, for instance the 2005 London underground bombings, the suicide bomb attack during a concert in Manchester, and the 2015 Paris attacks. Unfortunately the components required for building these devices are commercially available and the bombs relatively easy to construct.

Many explosives leave a characteristic trace after being handled or detonated, and it is essential that investigators can rapidly identify the components used in homemade explosives. Furthermore, the ability to trace the explosives back to particular individuals and terrorist cells is essential in preventing future attacks. Unfortunately effectively detecting and tracing explosives and explosive precursors can prove difficult. On top of this, after the production and implementation of IEDs, it can be difficult to prove a suspects’ involvement in bomb construction.

Researchers at King’s College London and Northumbria University have been working on developing new ways to detect homemade explosives.

The newly developed approach, recently published in the journal Analytica Chimica Acta, uses a technique known as ion chromatography high resolution mass spectrometry (IC-HRMS) to separate and detect chemical components. By applying the technique to compounds commonly encountered in the analysis of explosive residues, the method was shown to be effective in detecting a large number of components used to make bombs, capable of detecting just trace amounts of the chemicals faster than previous techniques.

Upon developing this method, the team of researchers then demonstrated that the approach could be applied to the analysis of human sweat, with the aim of indicating an individual has recently handled explosives. Participants were made to handle Pyrodex powder, a black powder propellant used in firearm cartridges. After handling the powder for a few minutes, palm sweat and fingermark samples were then collected at numerous timepoints over several hours. Analytes related to the explosive material were readily detected using the method. The real-world implementation of this technique could help prove contact between a suspect and explosive material or explosive precursors.

Gallidabino et al. Targeted and non-targeted forensic profiling of black powder substitutes and gunshot residue using gradient ion chromatography – high resolution mass spectrometry (IC-HRMS). Analytica chimica acta. 2019, In Press.

October 22, 2018 by Locard's Lab

Drug Detection at Your Fingertips: Illicit Drugs in Fingerprint Sweat

Researchers have developed a new tool for the rapid detection of a number of illicit drugs using only the sweat of an individual’s fingerprint.

Typically, the procedure to test for drugs in human beings necessitates the collection of blood or urine and laboratory-based analysis by gas or liquid chromatography with mass spectrometry. Unfortunately these standard methods are somewhat invasive, require potentially time-consuming laboratory-based analysis, and use complex pieces of analytical instrumentation requiring a trained operator to use. They are inevitably unsuitable for rapid, in-situ screening of potential drug users.

Researchers at the University of East Anglia and Intelligent Fingerprinting Ltd (a spin-out company from the university) have been working on a method of conducting simple, rapid drug analysis using sweat from a person’s finger. The technique has been developed to detect four classes of drugs – cannabis, cocaine, amphetamines and opiates, with cannabis being detected based on the presence of Δ⁹-tetrahydrocannabinol (THC), cocaine on the presence of benzoylecgonine, and opiates via the detection of morphine.

The finger of an individual is firmly pressed onto the Drug Screening Cartridge. This is then filled with a buffer solution before insertion into the reader for analysis. Capable of detecting drugs down to the picogram level, the system is a fluorescence-based lateral flow competition assay containing four drug-bovine serum albumin conjugate lines on a nitrocellulose test strip. In short, when a sample is introduced to the test strip, fluorescently-tagged antibodies pass over the conjugate lines. As these antibodies are specific to each drug class of interest, if that drug is present they will bind to the drug. At the end of the test, a fluorescence signal is measured. If none of the four drug classes were present, a maximum fluorescence signal will be obtained. However if any drugs were present to bind with the antibodies, there will be a decrease in the fluorescence signal proportional to the drug concentration. Within about 10 minutes, the device then gives a simple pass/fail response, requiring no specialist knowledge or excessive training to operate and interpret the results.

Furthermore, the technique has also been demonstrated to be effective when applied to the deceased. Researchers worked with a number of UK-based coroners to obtain fingerprint sweat samples from 75 deceased individuals. The most common drug detected was opiates, which is a logical finding considering the number of terminally ill patients who are prescribed morphine during palliative care.

In order to compare the new technique with those typically employed in the detection of drugs in human beings, analysis of blood samples was conducted by LC-MS-MS. The results between the two methods correlated well, with the accuracy between DSC of fingerprints and LC-MS-MS of blood being 88-97%, depending on the drug. This demonstrates the effectiveness of the method and its ability to stand up to existing techniques, though there are inevitably some shortcomings. Authors of the study have stated that there are known accuracy issues with lateral flow measurement devices, thus this new technology should be used as a presumptive screening method prior to confirmation by mass spectrometry. Furthermore, the range of target drugs is clearly currently limited, though future development could no doubt enable other classes of drug to be included.

Full details of the findings can be found in the Journal of Analytical Toxicology.

References

Hudson, T. Stuchinskaya, S. Ramma, J. Patel, C. Sievers, S. Goetz, S. Hines, E. Menzies and D. A. Russell, J. Anal. Toxicol., 2018, 6–10.

Forensic Magazine. Fingerprint Drug Screen Test Works on the Living and Deceased. [Available online] https://www.forensicmag.com/news/2018/10/fingerprint-drug-screen-test-works-living-and-deceased

August 9, 2017 by Locard's Lab

Tracking Illicit Drugs with Strontium Isotope Analysis

The manufacture and distribution of illicit drugs such as heroin is a primary focus of many major law enforcement organisations worldwide, including the Drug Enforcement Agency (DEA) in the United States and the National Crime Agency (NCA) in the United Kingdom. Unfortunately, as drug shipments pass hands between dealers and cross borders so rapidly, it can be difficult if not impossible to trace a batch of drugs back to an initial manufacturer. As a result of this, the chances of locating and arresting the manufacturers of illicit drugs can be slim.

To a forensic drugs analyst, a whole range of characteristics can be examined and used to classify and compare different batches of the same drug, including physical appearance, packaging, and chemical composition. To an extent, heroin chemical signatures are already beneficial in comparing different batches of the drug in attempts to establish links and possible sources of the narcotics. This may be based on agents or adulterants a product has been cut with, and the relative concentrations of those substances. The manufacturing process itself can vary in terms of chemicals and apparatus used and the skills of the manufacturer, resulting in further characteristic differences in the chemical profile. However these differences may not be distinct enough to be valuable and are certainly not able to pinpoint the country from which a batch originated. Though there is still no reliable method of tracing an illicit drug back to a particular location, ongoing research is aiming to change this.

One method of studying the history and even origin of a sample is to use isotopic composition. Isotopes are different forms of elements that are incorporated into substances in the environment in varying ratios and abundances, influenced by a number of factors that can alter these ratios. These processes can be described as isotopic fractionation. Interestingly, isotopic ratios can be characteristic to different regions of the world, enabling certain materials to be traced back to the geographic region based on the ratios of particular isotopes contained within that material. With this in mind, they have often been used to trace unidentified human remains to a particular location or study the origin of food products. Focusing on isotopes allows for heroin samples to be studied and compared based on regional characteristics as oppose to the variation caused by the production process.

For the first time, researchers at Florida International University have utilised strontium isotope ratio analysis to determine the provenance of illicit heroin samples. 186 unadulterated, undiluted heroin samples of known origin were obtained from a number of geographic regions including Southeast Asia, Southwest Asia, South America, and Mexico. Of a particularly challenging nature is South American heroin and SA-like Mexican heroin, which can be extremely difficult to differentiate based on their chemical compositions alone. Heroin samples were dissolved via a microwave-assisted acid digestion method before being subjected to a technique known as a multi-collector inductively-coupled plasma mass spectrometry (MC-ICP-MS). This instrument utilises an inductively coupled plasma ion source to ionise target analytes, which are then separated and analysed by the mass spectrometer. The use of MC-ICP-MS allows for the strontium concentration of particular samples to be determined. The strontium isotope ratio (⁸⁷Sr/⁸⁶Sr) value of each individual sample was then compared with the overall mean values of ratios from different regions in order to establish the likely origin of that particular heroin sample. Samples from the same geographic region would be expected to exhibit a similar isotope ratio.

Multi-collector inductively-coupled plasma mass spectrometer (MC-ICP-MS) Source: www.thermofisher.com

The results demonstrated the possibility of differentiating between heroin of different geographic origin. South American and Mexican heroin samples were correctly classified 82% and 77% of the time respectively. South East and South West Asian heroin samples were somewhat more difficult to differentiate due to more of an overlap between strontium isotope ratio values. SE Asian samples were correctly classified 63% of the time and SW Asian samples only 56% of the time. It is not clear whether this elemental strontium is endogenous or the result of external contamination, but either way it is sufficiently characteristic to be associated with a particular geographic location.

The strontium isotope composition of heroin can be affected by a number of factors, including the soil in which components are grown and groundwater in the area, which can result in region-specific isotope compositions. The use of strontium isotope ratio analysis has presented promising results in the origin determination of illicit heroin. Although a larger scale study incorporating samples of a more worldwide origin would be ideal, initial results suggest that this technique could allow for an unknown illicit drug sample to be traced back to a country of origin, aiding criminal intelligence agencies in the war against drugs.

Debord, J., Pourmand, A., Jantzi, S., Panicker, S. & Almirall, J. Profiling of Heroin and Assignment of Provenance by ⁸⁷Sr/⁸⁶Sr Isotope Ratio Analysis. Inorg Chim Acta. In press (2017).

May 15, 2017 by Locard's Lab

Keeping the Skies Safe with Analytical Chemistry

Ever since events such as 9/11, the Lockerbie bombing and the (fortunately) failed shoe bomber, the stringency of airport security has been ever increasing. Anyone who has passed through an airport has no doubt witnessed the occasional swabbing of luggage or electronic items. The staff will take a quick swab of the item, stick it into a mysterious machine and usually send the passenger on their way with little explanation of what has just occurred.

But what exactly are they testing for in this scenario, and just what is the instrument they’re using?

As one might expect, the biggest target of this security step is explosive substances as a counter-terrorism measure, in addition to illicit narcotics in an attempt to crack down on drug trafficking. In an airport setting, the analytical testing technique of choice is ion mobility spectrometry.

Ion mobility spectrometry (IMS) is an analytical technique used to identify chemical compounds based on the differences in the movement of ions under an electric field. The concept for the technique was established in the early twentieth century, however it was not until the 1970s that the instrumentation was actually properly developed. There are currently tens of thousands of IMS devices deployed around the world. Not only are they utilised in airports for drug and explosives screening, but also by the military for the detection of chemical warfare agents and in industrial settings to monitor air quality. The range of applications is potentially vast, but the principles of operation are the same.

As you may have witnessed, a small swab is rubbed over the surface to be tested, typically a piece of luggage or an electronic device such as a laptop, before being inserted into the ion mobility spectrometer. As the sample needs to be introduced in its gaseous form, the swab may be subjected to heating in order to thermally desorb analytes from the swab and allow them to be transported into the instrument for analysis. In order to manipulate the analytes entering the instrument, they must first be converted into ions, their charged form. Ionisation is typically achieved using a radioactive source, such as ⁶³Ni (nickel-63) or ²⁴¹Am (Americium-241), which first form reactant ion species from the carrier gas (usually air), which then leads to the ionisation of the sample material. These newly-formed ions will then enter a region under an electric field and drift towards a series of electrodes. The ions will pass through the drift region at different speeds depending on the shape and size of the ion clusters and strike the electrodes, the signals being amplified and detected. Depending on the instrument and needs of the analysis, either positive or negative ions will be produced (in some cases both simultaneously).

IMS schematic. Source: Smiths Detection (www.smithsdetection.com)

The IMS utilised in airports will typically hold a database of known explosive and narcotic substances against which to compare samples. There will be a certain threshold, typically based on peak intensity, that must be reached before a positive identification will be indicated, and if there is a “match”, the operator will be alerted to a potential identification.

In comparison to other analytical tools available, ion mobility spectrometers are far from being the best. For instance mass spectrometry, an alternative technique for the analysis and identification of chemical compounds, can offer greater sensitivity, higher resolution, improved accuracy and better identification. So why use IMS? It essentially comes down to cost and ease of use. The simple design and ability to operate at atmospheric pressure means the instruments can be fairly small in size, some even being hand-held and so rendering them completely portable. They have low power consumption, so can simply be powered by a few AA batteries. The ease of use of the IMS means anyone can be trained to use the instrument, thus technical or scientific expertise is not required.

But what is perhaps most important for use in an airport setting with potentially thousands of passengers each hour, is the ability to conduct analyses quickly, and this is something that the IMS can offer. Many commercial ion mobility-based instruments can provide results in a matter of seconds. For instance, the IONSCAN by Barringer (now owned by Smiths Detection) boasts the ability to detect over 40 explosives and narcotics in just 8 seconds.

In a security setting there are three primary types of IMS that may be encountered. The smallest of the devices are handheld and sample by drawing in analytes present in the atmosphere. These may be used to analyse potential hazards relating to unattended baggage, for example. The second type, which is perhaps the most commonly encountered IMS in airports, is a benchtop instrument which requires introduction of the sample via some type of swab. And finally, some airport security units may utilise a larger, human-sized IMS portal. This setup uses airflow to dislodge particles of explosives or drugs from clothing or the passenger’s body and analyse them.

Unsurprisingly, the instruments are not infallible, and false positive or negative results are a possibility. Some ions will have the same drift time so may be indistinguishable from known explosives or drugs, triggering an alarm. In actual fact this response may simply have been caused by a cosmetic or pharmaceutical product that happens to produce a response similar to a known narcotic. On the contrary, dirt, oil and other contaminants may mask the presence of substances of interest, thus causing no alert despite the presence of a drug or explosive.

Furthermore, the IMS is somewhat limited in that it can only identify the presence of a compound contained within its database. So whereas it may be able to detect common explosives such as RDX, TNT and PETN, and frequently encountered narcotics such as cocaine, heroin and cannabis, it would not necessarily alert to the presence of an unknown compound (unless it was very similar in chemical structure to something in the database).

Fortunately research in the field of analytical chemistry is constantly ongoing, aiming to improve instrumentation and analytical techniques to resolve these issues and ultimately produce more reliable and robust security measures.

References

G. Ewing et al. A critical review of ion mobility spectrometry for the detection of explosive and explosive related compounds. Talanta. 54 (2001) 515-529.

Homeland Security Science & Technology. IMS-Based Trace Explosives Detectors for First Responders. [online] Available: https://www.dhs.gov/sites/default/files/publications/IMSTraceExploDetect-SUM_0506-508.pdf

Smiths Detection. Ion Mobility Spectrometry (IMS). [online] Available: https://www.smithsdetection.com/index.php?option=com_k2&view=item&layout=item&id=40&Itemid=638

January 24, 2017 by Locard's Lab

Sex Determination with Raman Spectroscopy

The ability to quickly identify a victim or suspect during a criminal investigation is crucial, and the use of fingerprinting and DNA profiling often proves invaluable in this. However, a fingerprint or DNA profile can only be associated with an individual if there is an alternative profile or database match for comparison.

But what can investigators do when comparison profiles are not available, rendering biological fluids found at crime scenes somewhat useless?

The capability of instantly establishing alternative information relating to a suspect – such as sex, age or a phenotypic characteristic – based on the analysis of the evidence could be a substantial benefit to an investigation.

In recent years, the use of both well-established and novel analytical techniques to ascertain information relating to a suspect or victim from bodily fluids has been the focus of a great deal of research. With an increasing number of analytical instruments becoming field portable, the possibility of in situ analysis at crime scenes and instant suspect information is quickly becoming a reality.

Raman Spectroscopy and Sex Determination

Most recently, researchers at the University of Albany (Muro et al, 2016) have highlighted the possibility of using portable Raman Spectroscopy to determine the sex of an individual based only on their saliva in real-time.

The study utilised a total of 48 saliva samples from both male and female donors of multiple ethnicities, depositing the samples onto aluminium foil and drying overnight. Samples were then subjected to Raman analysis and the chemical signatures scrutinised to determine whether or not the saliva of male donors differed from that of female donors.

Raman Spectroscopy is a non-destructive analytical technique used for analyte identification based on molecular vibrations. As a basic explanation, monochromatic light is initially directed towards the sample, some of this light simply passing through the sample and some of it being scattered. A small amount of this scattered light experiences an energy shift due to interactions between the sample and the incident light. These energy shifts are detected and transformed into a visual representation. The resulting Raman spectrum typically plots frequency vs intensity of the energy shifted light. The positions of different bands on this spectrum relate to the molecular vibrations within the sample which, if interpreted correctly, can allow for the identification of analytes.

Raman spectra are somewhat characteristic of the chemical composition of the sample. In the case of the saliva analysed in this study, the features of the spectra were largely caused by amino acids and proteins. When comparing the respective spectra from male and female donors, by eye they appear remarkably similar. However using multivariate data analysis, a statistical technique used to analyse data with multiple variables, the researchers were able to distinguish between the saliva of male donors and that of female donors, reporting the ability to ascertain the sex of the donor with an accuracy of an impressive 94%.

Comparison of male and female saliva Raman spectra (Muro et al, 2016)

Although only a proof-of-concept paper, the research demonstrates the possibility of using portable Raman spectroscopy as a method of elucidating donor information, in this case sex, through the analysis of a bodily fluid. The researchers suggest further work will be conducted to include other bodily fluids and donor characteristics.

At this point, the usefulness of the research is limited. Although instantly establishing the sex of the donor of a bodily fluid can aid investigators in developing a suspect or victim profile more efficiently, the pool of potential donors is still huge. The total of 48 saliva donors used in this study is of course not a sufficient representation of the population, thus a much larger sample set would be required to fully evaluate the technique, including non-laboratory setting experiments. Furthermore, there is a wide range of medical conditions and additional factors that can result in changes in the chemical composition of saliva and thus could influence the effectiveness of this technique. Whether or not certain diseases or external influences can hinder gender determination using this method would need to be investigated.

Previous Research

The idea of utilising analytical chemistry to ascertain donor information is not in itself novel, and other researchers have attempted to achieve the same goal through different means.

In 2015, scientists also based at the University of Albany (Huynh et al, 2015) developed a biocatalytic assay approach to the analysis of amino acids in fingerprints to determine the sex of the donor. The study boasted an accuracy of 99%, with the sex differences believed to be due to the higher concentration of amino acids in fingerprints deposited by females.

Research by Takeda et al in 2009 used Nuclear Magnetic Resonance (NMR) Spectroscopy to determine differences between the urine and saliva samples of different donors based on the detection and comparison of different metabolites. Certain compounds, including acetate, formate, glycine and pyruvate, were found in higher concentrations in male samples, allowing for the differentiation between male and female bodily fluids.

The focus of such research is not limited to sex differentiation, for instance some research has even focused on establishing whether a blood sample belongs to a smoker or non-smoker. Utilising gas chromatography mass spectrometry with a solid phase microextraction pre-concentration step, Mochalski et al (2013) were able to effectively distinguish between the blood and breath of smokers and non-smokers due to the ten-fold increase in levels of benzene and toluene, a conclusion which has been repeated by other researchers.

Looking at just this small handful of studies, it becomes evident that certain analytical techniques have the potential power to ascertain a range of information about the donor of a bodily fluid. However all of these immunoassay and mass spectrometry techniques are typically time-consuming, requiring the transportation of a sample to a laboratory, sometimes extensive sample preparation, followed by a form of analysis that will often destroy the sample. This is evidentially not ideal during a time-sensitive criminal investigation in which sample amount may be limited.

To an extent, the research utilising Raman spectroscopy to determine sex from saliva does alleviate some of these problems. The portability of Raman devices allows for in situ analysis, removing the need for expensive and time-consuming laboratory analysis. As Raman spectroscopy is based on the interaction between the sample analyte and light, it is a non-destructive technique, allowing the sample to be preserved for storage and further analyses is required.

Although these techniques do not hold the power of DNA in almost irrefutably identifying the suspect, they may at least aid investigators in narrowing down their pool of suspects and steering the investigation in the right direction. No doubt further advances in analytical chemistry will allow for more accurate and robust techniques in the future.

References

Huynh, C et al. Forensic identification of gender from fingerprints. Anal. Chem. 87(2015), pp11531-11536.

Mochalski, P et al. Blood and breath levels of selected volatile organic compounds in healthy volunteers. Analyst. 7(2013), pp2134-2145.

Muro, C. L et al. Sex determination based on Raman Spectroscopy of saliva traces for forensic purposes. Anal. Chem. 88(2016), pp12489-12493.

Takeda, I et al. Understanding the human salivary metabolome. NMR Biomed. 22(2009), pp577-584.