Metabolism of Carfentanil, an Ultra-Potent Opioid, in Human Liver Microsomes and Human Hepatocytes by High-Resolution Mass Spectrometry.

Carfentanil is an ultra-potent synthetic opioid. No human carfentanil metabolism data are available. Reportedly, Russian police forces used carfentanil and remifentanil to resolve a hostage situation in Moscow in 2002. This alleged use prompted interest in the pharmacology and toxicology of carfentanil in humans. Our study was conducted to identify human carfentanil metabolites and to assess carfentanil's metabolic clearance, which could contribute to its acute toxicity in humans. We used Simulations Plus's ADMET Predictor™ and Molecular Discovery's MetaSite™ to predict possible metabolite formation. Both programs gave similar results that were generally good but did not capture all metabolites seen in vitro. We incubated carfentanil with human hepatocytes for up to 1 h and analyzed samples on a Sciex 3200 QTRAP mass spectrometer to measure parent compound depletion and extrapolated that to represent intrinsic clearance. Pooled primary human hepatocytes were then incubated with carfentanil up to 6 h and analyzed for metabolite identification on a Sciex 5600+ TripleTOF (QTOF) high-resolution mass spectrometer. MS and MS/MS analyses elucidated the structures of the most abundant metabolites. Twelve metabolites were identified in total. N-Dealkylation and monohydroxylation of the piperidine ring were the dominant metabolic pathways. Two N-oxide metabolites and one glucuronide metabolite were observed. Surprisingly, ester hydrolysis was not a major metabolic pathway for carfentanil. While the human liver microsomal system demonstrated rapid clearance by CYP enzymes, the hepatocyte incubations showed much slower clearance, possibly providing some insight into the long duration of carfentanil's effects.

An Impedance-Based Model for the Assessment of Cardiopulmonary Function in Rabbits.

Improving the quality of physiologic data collected from research animals is most easily accomplished by collecting as much information as possible from a single subject, thereby reducing animal use and error associated with satellite groups. We investigated the feasibility of using a large-animal implantable telemetry device in New Zealand white rabbits (n = 6). The first task was to develop an implantation technique that yielded calibrated tidal volume (Vt) measurements that were within 10% of those obtained simultaneously from a pneumotachograph, a low-noise electrocardiogram, and stable blood pressure. The second task was to challenge implanted rabbits with the respiratory stimulant doxapram to assess linearity of the calibration across a range of Vt. Of the 3 electrode placements attempted, only one resulted in calibrations consistently below 10% error. Optimal electrode placement resulted in calibrated Vt measurements within 1.7% ± 0.3% of those obtained from a pneumotachograph during normal tidal breathing, 7.3% ± 0.7% of those after saline injection, and 6.0% ± 0.5% of those after doxapram injection. The Vt range was 9 to 15 mL for normal tidal breathing and saline injection and 25 to 30 mL after doxapram injection. Increases in mean arterial pressure of 25.0 ± 6.82 mm Hg and decreases in heart rate of 56.3 ± 6.82 bpm were associated with doxapram injection only. Our findings represent the first time that multiple cardiopulmonary endpoints have been assessed by telemetry in conscious, restrained rabbits. Whether animal position affects calibration accuracy warrants investigation.

Nitration of benzo[a]pyrene adsorbed on coal fly ash particles by nitrogen dioxide: role of thermal activation.

Nitration of benzo[a]pyrene (BaP) by nitrogen dioxide (NO2) adsorbed on the surface of thermally activated coal fly ash and model aluminosilicate particles led to the formation of nitrobenzo[a]pyrenes as verified by extraction and gas chromatography/mass spectrometry (GC/MS). In situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) was utilized to follow the nitration reaction on the surface of zeolite Y. Nitrobenzo[a]pyrene formation was observed along with the formation of nitrous acid and nitrate species. The formation of the BaP radical cation was also observed on thermally activated aluminosilicate particles by electron spin resonance (ESR) spectroscopy. On the basis of GC/MS, DRIFTS, and ESR spectroscopy results, a mechanism of nitration involving intermediate BaP radical cations generated on thermally activated aluminosilicate particles is proposed. These observations have led to the hypothesis that nitration of adsorbed polyaromatic hydrocarbons on coal fly ash by reaction with nitrogen oxides can occur in the smokestack, but with the aging of the fly ash particles, the extent of the nitration reaction will be diminished.