In recent years, an increased number of reports concerning illegal production and non-medical use of fentanyl and its analogues found in NPS samples have appeared. Because of high potency, these compounds are used in very low doses and most of them are highly dangerous and toxic. Forensic quantitative analysis of seized NPS samples is requested by the law enforcement administration (police,
... [Show full abstract] judges, prosecutors) usually in order to estimate the number of active doses and thus the number of people who can be intoxicated. It can be helpful in estimating whether they deal with a large scale case or minor one. This number of doses is taken into account when proceeding a forensic case and passing a sentence in court. To perform such an analysis, the forensic laboratory should use individual quantitative reference standards. In case of NPS such compounds may sometimes be commercially unavailable. Continuing purchase of NPS standards is also very expensive, and the time needed to obtain controlled drugs may be very long. All these were the reasons why the authors searched for an alternative method of NPS quantification in such instances. Application of charged aerosol detection (CAD) combined with ultra-performance liquid chromatography (UHPLC) as a separation technique, appeared to be such a solution. Instead of individual reference standards of the analytes, the method utilises only one certified standard called the reference compound in this case para-fluorofentanyl. Quantification of the detected substances is then based on the calibration curve of the chosen reference compound – para-fluorofentanyl. This procedure can be used for determination of fentanyl analogues in the forensic analysis of seized NPS without the use of individual reference standards.