Semi‐quantitative analysis of tramadol, dextromethorphan, and metabolites in decomposed skeletal tissues by ultra performance liquid chromatography quadrupole time of flight mass spectrometry

Abstract

The use of filtration/pass-through extraction (FPTE) and ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-qTOF-MS) to detect tramadol (TRAM), dextromethorphan (DXM), and metabolites from skeletal remains is described. Rats (n=5) received 50 mg/kg tramadol and were euthanized by CO asphyxiation approximately 30 minutes post-dose. Rats (n=4) received 75 mg/kg dextromethorphan and were euthanized by CO asphyxiation approximately 45 minutes post-dose. Remains decomposed to skeleton outdoors and vertebral bones were collected. Bones were cleaned, dried, and pulverized to a fine powder. Bones underwent dynamic methanolic extraction followed by FPTE before analysis using UPLC-qTOF-MS. Recovery was at least 90% of maximal value within the first 10 minutes of methanolic extraction for all samples assayed. Analytical response was measured over the concentration range of 1-500 ng/mL, with precision and bias <20% in triplicate analyses of all calibrators, and a limit of detection of 1 ng/mL for TRAM, DXM, and all metabolites. The vertebral bone analyzed using this method detected TRAM, DXM, and their respective metabolites in all samples analyzed.

Development and Validation of a Method for the Analysis of Bisoprolol and Atenolol in Human Bone

“…The RR/m ratios are found proportional to the concentration in the validation protocol, so drugs levels can be compared with bone samples prepared in the same way over the range of concentrations assessed in the validation protocol. For better understanding, RR/m values have been expressed in concentration units (ng/mg), but they should only be used as approximations [7,12].…”

“…In case of significant putrefaction, skeletal tissues may provide the only source of information, since these specimens are well protected from decomposition [2,3,4,5]. In the last decade, skeletal tissues have been investigated as postmortem toxicological matrix by different authors who used a variety of methodologies for drugs extraction and detection [6,7,8,9,10,11,12,13,14,15,16]. Most researchers used experimental animals with the advantage of working under controlled conditions [2,4,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21], while studies performed using human bones [1,5,22,23,24,25,26,27,28,29] are scarce and to date, do not use standardized protocols for sample preparation and analysis [2,28,29].…”

Recent advances in nanomaterials-based electrochemical sensors for tramadol analysis

“…These methods are commonly used in biology for research, development and quality control of pharmaceutical products. Quantitative determination of tramadol in pharmaceutical and biological samples has been investigated by some instrumental methods such as gas chromatography (GC) [ 16 ] gas chromatography-mass spectrometry (GC-MS) [ 17 , 18 ], high-performance liquid chromatography (HPLC) [ 19 ], capillary electrophoresis [ 20 , 21 ], spectrophotometry [ 22 , 23 ], chemiluminescence [ 24 ] and electrochemical methods [ 25 ]. However, most of these methods require highly trained technicians and complex sample preparation that hinders their wide application.…”