Mass Spectrometry & Spectroscopy
Tackling the Chromatographic Analysis of Novel Psychoactive Substances with High Resolution Mass Spectrometry
Phil Taylor Global Marketing Manager, Food, Environmental and Forensic at Sciex, Phoenix House, Lakeside Drive, Warrington, Cheshire, WA1 1RX, UK, Email:
philip.Taylor@sciex.com
On the 26th of May, 2016 the United Kingdom introduced the ‘Psychoactive Substances Act 2016’ [1], this law is intended to restrict the production, sale and supply of novel psychoactive substances (NPS) or as they are more commonly known, legal highs or designer drugs.
This law defi nes a ‘psychoactive substance’ as anything, which by stimulating or depressing the person’s central nervous system affects the persons mental functioning or emotional state.
In November 2016 Germany also introduced ‘The New Psychoactive Substances Act’ (NpSG) [2]. This law prohibits the acquisition, possession and sale of new psychoactive substances (NPS) as well as sanctioning the passing on of NPS. The NpSG specifi cally controls over two NPS groups; they are phenethylamines and synthetic cannabinoids.
As governments around the world make new laws to combat the production of the NPS, the effects of these substances are still real. Announcements in November 2016 on the ‘UNODC Early Warning Advisory on New Psychoactive Substances’ [3] documents increasing numbers of serious adverse events and deaths associated with several emerging synthetic opioids.
Authorities need to be able to rapidly detect the NPS with confi dence and accuracy to tackle the problem. However, having the reference standard to confi rm the presence of NPS is often diffi cult if the chromatography and analytical instrumentation such as a mass spectrometer rely on already knowing the details of specifi c compounds. This includes information such as retention time (RT), their mass to charge ratio (m/z) and the multiple reaction monitoring (MRM) transitions. Advanced systems that are accompanied by libraries which deliver high quality spectral data to confi rm a known NPS, and contain these details (RT, m/z, MRM and libraries) are vital if you wish to conduct a screening assay using LC-MS/MS for example.
However, the NPS situation constantly evolves, as the clandestine laboratories that manufacture them make modifi cations to the structure (analogues), stereoisomers and derivatives. In doing this, the NPS may increase potency and alter the side effects of the designer drug. Once the NPS has been modifi ed, it becomes diffi cult if not impossible to analyse.
In this study 85 analytes were selected which would challenge chromatographic separation and test the instrument to detect and screen the compounds in an ultra-fast time frame. Although in a real-life case it would be highly unlikely that any sample would contain this many forensic drugs.
Experimental
In a recent application note ‘Ultra-Fast Forensic Toxicological Screening and Quantitation under 3 Minutes using Sciex X500R QTOF System and Sciex OS 1.0 Software’ [4] novel psychoactive substances were addressed in the study of urine samples. Twenty samples were diluted in 10% methanol and centrifuged to reduce the possibility of matrix interference. The clear supernatants were transferred to autosampler vials which were spiked with 85 compounds including opioids and synthetic cannabinoids. The urine samples contained a calibrator set spiked with compounds described in Table 1. There are 7 levels of calibrators: 40%, 80%, 100%, 200%, 300%, 500% and 1000% which are against the ng/mL value of each compound in table one, for example for the compound of 6-MAM, a metabolite of heroin, the calibration range was 4ng/mL to 100ng/mL. Using such a calibration range enable low level detection, sub cut off level through to a high level of concentration at 100ppb. There were also 20 urine samples with unknown number of compounds. The injection volume was 5 µL with a dilution factor for this sample set was a x4, again this reduces matrix interferences and the need for instrument maintenance.
Figure 1. LC Gradient conditions
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