Green aspects in the procedure of detection ketamine, flunitrazepam, and diazepam in drinks based on dried sample spot analysis

A new methodology for the detection of ketamine, flunitrazepam, and diazepam in beverage samples by the dried sample spot method was developed. The method is characterized by very low LODs in all tested types of beverages (100 ng/mL for ketamine, 25 ng/mL for flunitrazepam and diazepam) and great precision at the concentration of 100 ng/mL for all analytes. The significant advantages of this method are the consumption of fewer amount of samples and the possibility of securing the beverage samples on DBS cards at room temperature. The proposed method was evaluated by the innovative WAC approach according to Green Analytical Chemistry. The results of the evaluation indicate the best results of this method in terms of analytical quality compared to the other methods from the literature, however other aspects such as green chemistry and economical are also good. The DBS/MAE/LC-MS method could be used for qualitative analysis for drugs detection in cases of date-rape drugs analysis

The double face of ketamine : the possibility of its identification in blood and beverages

The purpose of this study was to develop and validate a high-sensitivity methodology for identifying one of the most used drugs—ketamine. Ketamine is used medicinally to treat depression, alcoholism, and heroin addiction. Moreover, ketamine is the main ingredient used in so-called “date-rape” pills (DRP). This study presents a novel methodology for the simultaneous determination of ketamine based on the Dried Blood Spot (DBS) method, in combination with capillary electrophoresis coupled with a mass spectrometer (CE-TOF-MS). Then, 6-mm circles were punched out from DBS collected on Whatman DMPK-C paper and extracted using microwave-assisted extraction (MAE). The assay was linear in the range of 25–300 ng/mL. Values of limits of detection (LOD = 6.0 ng/mL) and quantification (LOQ = 19.8 ng/mL) were determined based on the signal to noise ratio. Intra-day precision at each determined concentration level was in the range of 6.1–11.1%, and inter-day between 7.9–13.1%. The obtained precision was under 15.0% (for medium and high concentrations) and lower than 20.0% (for low concentrations), which are in accordance with acceptance criteria. Therefore, the DBS/MAE/CE-TOF-MS method was successfully checked for analysis of ketamine in matrices other than blood, i.e., rose wine and orange juice. Moreover, it is possible to identify ketamine in the presence of flunitrazepam, which is the other most popular ingredient used in DRP. Based on this information, the selectivity of the proposed methodology for identifying ketamine in the presence of other components of rape pills was checked

Nitrazepam and 7-aminonitrazepam studied at the macroscopic and microscopic electrified liquid-liquid interface

Two benzodiazepine type drugs, that is, nitrazepam and 7-aminonitrazepam, were studied at the electrified liquid-liquid interface (eLLI). Both drugs are illicit and act sedative in the human body and moreover are used as date rape drugs. Existence of the diazepine ring in the concerned chemicals structure and one additional amine group (for 7-aminonitrazepam) allows for the molecular charging below their pKa values, and hence, both drugs can cross the eLLI interface upon application of the appropriate value of the Galvani potential difference. Chosen molecules were studied at the macroscopic eLLI formed in the four electrode cell and microscopic eLLI formed within a microtip defined as the single pore having 25 μm in diameter. Microscopic eLLI was formed using only a few μL of the organic and the aqueous phase with the help of a 3D printed cell. Parameters such as limit of detection and voltammetric detection sensitivity are derived from the experimental data. Developed methodology was used to detect nitrazepam in pharmaceutical formulation and both drugs (nitrazepam and 7-aminonitrazepam) in spiked biological fluids (urine and blood).R. Wietecha-Posłuszny and P. Stelmaszczyk are grateful for the financial support (National Science Centre, Grant no. UMO-2019/35/O/ST4/00978: application of developed extraction procedure, study the interfacial behavior of NIT and 7a-NIT), so is L. Poltorak (National Science Centre, Grant no. UMO-2018/31/D/ST4/03259: application of designed fused silica capillary and 3D printed cell for NIT and 7a-NIT detection)

Optimized and Validated DBS/MAE/LC–MS Method for Rapid Determination of Date-Rape Drugs and Cocaine in Human Blood Samples—A New Tool in Forensic Analysis

The aim of this work was to develop a new method for the determination of selected substances from the date-rape drugs group: ketamine, benzodiazepines and cocaine. The method is based on the dried blood spot method which seems to be a suitable tool in the analysis of tested substances. The extraction process based on microwave-assisted extraction was optimized to enable optimal conditions for the isolation of a wide range of analytes from blood samples collected on DBS cards. The extraction with ethyl acetate with a buffer of pH = 9 carried out at a temperature of 50 °C for 15 min ensured high extraction efficiency of the tested analytes. The optimized method was validated. Limits of detection (LOD = 4.38–21.1 ng/mL) and quantification (LOQ = 14.6–70.4 ng/mL), inter- and intra-day precision (CV = 1.37–13.4% and 3.39–14.8%, respectively), recovery (RE = 93.0–112.4%) and matrix effect (ME = 98.4–101.6%) were determined. The validation results indicate the possibility of using the proposed method in the analysis of real blood samples collected from victims of sexual assault

High-performance liquid chromatography method for analysis of pharmaceutical preparations

Niniejsza praca przedstawia zastosowanie wysokosprawnej chromatografii cieczowej do analizy wybranych preparatów farmaceutycznych pod kątem zawartości acetaminofenu i kofeiny. W celu identyfikacji substancji zastosowano detektor z matrycą diodową (DAD) połączonego z chromatografem cieczowym. Kalibrację oznaczenia wykonano metodą wzorca wewnętrznego, przy użyciu kwasu acetylosalicylowego. Jako fazę ruchomą użyto mieszaniny fazy wodnej o pH = 2,4 i acetonitrylu w stosunku objętościowym 85:15. Wyznaczone parametry walidacyjne: liniowość (R2 = 0,9994 dla acetaminofenu oraz R2 = 0,9995 dla kofeiny), granica wykrywalności (LOD: 4 ∙ 10-5 mg/mL dla acetaminofenu, 10-4 mg/mL dla kofeiny), granica oznaczalności (LOQ: 1,2 ∙ 10-4 mg/mL dla acetaminofenu, 3 ∙ 10-4 mg/mL dla kofeiny), precyzja pomiarów podczas jednego dnia (współczynnik zmienności CV%: 0,58 – 0,67% dla acetaminofenu, 0,96 – 1,66% dla kofeiny), precyzja pomiarów między dniami (współczynnik zmienności CV%: 1,75 – 2,39% dla acetaminofenu, 1,94 – 2,93% dla kofeiny), poprawność metody (RE%: -2,49 – 0,11% dla acetaminofenu, 3,17 – 5,87% dla kofeiny) oraz procent odzysku (99,89 – 102,49% dla acetaminofenu, 94,13 – 96,83% dla kofeiny) uznano za satysfakcjonujące. Opracowana metoda znajduje zastosowanie w jednoczesnym oznaczaniu zawartości acetaminofenu oraz kofeiny w preparatach farmaceutycznych.This paper presents application of the high performance liquid chromatography method in the analysis of pharmaceutical preparations containing acetaminophen and caffeine. In order to identify the substances it was used diode array detector (DAD) connected with the liquid chromatograph. Calibration of the assay was done by internal standard using acetylsalicylic acid. A mixture of aqueous phase having a pH of 2.4 and acetonitrile in a volume ratio of 85:15 was used as a mobile phase. The validation parameters: linearity (R2 = 0,9994 for acetaminophen, R2 = 0,9995 for caffeine), detection limit (LOD: 4 ∙ 10-5 mg/mL for acetaminophen, 10-4 mg/mL dla caffeine), quantitation limit (LOQ: 1,2 ∙ 10-4 mg/mL for acetaminophen, 3 ∙ 10-4 mg/mL for caffeine), intra-day precision (CV%: 0,58 – 0,67% for acetaminophen, 0,96 – 1,66% for caffeine), inter-day precision ( CV%: 1,75 – 2,39% for acetaminophen, 1,94 – 2,93% for caffeine), accuracy (RE%: -2,49 – 0,11% for acetaminophen, 3,17 – 5,87% for caffeine and the percent recovery (99,89 – 102,49% for acetaminophen, 94,13 – 96,83% for caffeine) were found to be satisfactory. The developed method is applicable in simultaneous determination of acetaminophen and caffeine in pharmaceutical preparations

Optimization and application of the DBS/CE-MS method for the analysis of human blood collected intravenously and post-mortem

Niniejsza praca opisuje optymalizację i wykorzystania metody DBS/MAE/CE-MS w analizie próbek krwi pobranych przyżyciowo lub pośmiertnie dla celów terapeutycznego monitorowania stężenia leków i toksykologii sądowej. Metoda ta umożliwia oznaczenie wybranych leków z grup TLPD, SSRI BZDs i leków nasennych, stosowanych w leczeniu zaburzeń nastrojów.W trakcie badań zoptymalizowano warunki separacji badanych leków przy użyciu techniki CE-MS i parametry ekstrakcji MAE analitów z krwi naniesionych na karty DBS typu FTA DMPK C. Metoda ta wymaga niewielkiej ilości próbki do analizy i generuje niewielką ilość odpadów, można więc ją uznać za metodę „zielonej chemii”. Ponadto metodę poddano procesowi walidacji. Wyznaczono wartości granic wykrywalności i oznaczalności badanych leków, precyzję oznaczeń w ciągu dnia (CV: 1,31 – 9,43%) oraz pomiędzy dniami (CV: 3,26 – 18,52%), odzysk (RE: 85,0 – 105,4%) i efekt matrycy na jonizację analitów (ME: 98,6 – 105,5%). Opracowana metoda jest selektywna, a anality w krwi naniesionej na karty DBS są stabilne w przeciągu co najmniej 14 dni.Wyniki analizy trzech próbek rzeczywistych wskazują na możliwość zastosowania metody DBS/MAE/CE-MS w analizie krwi pobranej przyżyciowo oraz pośmiertnie dla potrzeb monitorowania stężenia leków we krwi i toksykologii sądowej.This paper describes the optimization and application of the DBS/MAE/CE-MS method in the analysis of blood samples collected intravenously or post-mortem for therapeutic monitoring of drug concentration and forensic toxicology. This method allows to determine selected drugs from TLPD, SSRI, BZDs and hypnotics used in the treatment of mood disorders.During the research, the conditions of separation process of the tested drugs using the CE-MS technique and the parameters of MAE extraction of analytes from blood applied on FTA DMPK C DBS cards were optimized. This method requires a small amount of sample for analysis and generates a small amount of waste, so it can be considered as a “green chemistry” method. The method was subjected to the validation process. The limits of detection and quantification , the precision of the markings during the day (CV: 1,31 – 9,43%) and between days (CV: 3,26 – 18,52%), the recovery (RE: 85,0 – 105,4%) and matrix effect on ionization of analytes (ME: 98,6 – 105,5%) were determined. The developed method is selective and analytes in the blood applied on DBS cards are stable for at least 14 days.The results of the analysis of three real samples indicate the applicability of the DBS/MAE/CE-MS method in the analysis of the blood collected intravenously and post-mortem for the purposes of blood concentration monitoring and forensic toxicology

Glosa do wyroku Wojewódzkiego Sądu Administracyjnego w Gdańsku z dnia 17 września 2014 r., I SA/Gd 1632/13

Zajęciem na cele działalności gospodarczej gruntu wyłączonego z opodatkowania podatkiem od nieruchomości (gruntu rolnego) będą faktyczne czynności polegające na przeznaczeniu i  wykorzystywaniu gruntu do prowadzenia działalności, z zaznaczeniem, że wszelkie działania winny być uzasadnione celami gospodarczymi przedsiębiorcy. Kolejną konieczną przesłanką dla uznania gruntu rolnego za zajętego na potrzeby tej działalności będzie brak możliwości prowadzenia działalności rolnej