LC-MS-MS-MS3 for the determination and quantification of ∆9-tetrahydrocannabinol and metabolites in blood samples.

Due to the high prevalence of cannabinoids in forensic toxicology analysis, it is crucial to have an efficient method that allows the use of a small sample amount and that requires a minimal sample preparation for the determination and quantification of low concentrations. A simple, highly selective and high throughput liquid chromatography-tandem mass spectrometry methodology (LC-MS-MS-MS3) was developed for the determination and quantification of ∆9-tetrahydrocannabinol (THC), 11-hydroxy-∆9- tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-∆9-tetrahydrocannabinol (THC-COOH) in blood samples. Chromatographic analysis of THC, THC-OH and THC-COOH and their deuterated internal standards was preceded by protein precipitation (PPT) of 0.1 mL of blood samples with acetonitrile. Chromatographic separation was achieved by use of an Acquity UPLC® HHS T3 (100 mm × 2.1 mm i.d., 1.8 µm) reversed-phase column, using a gradient elution of 2 mM aqueous ammonium formate, 0.1% formic acid and methanol at a flow rate of 0.4 mL/min, with a run time of 10 min. For the MS-MS-MS3 analysis, a SCIEX QTRAP® 6500+ triple quadrupole linear ion trap mass spectrometer was used via electrospray ionization (ESI), operated in multiple reaction monitoring (MRM) and linear ion trap mode (MS3). The method was validated in accordance with internationally accepted criteria and guidelines, and proved to be selective and linear between 0.5 and 100 ng/mL (r2 > 0.995). The lower limits of quantification (LLOQ) corresponded to the lowest concentrations used for the calibration curves. The coefficients of variation obtained for accuracy and precision were <15%. The mean recoveries were between 88.0% and 117.2% for the studied concentration levels (1 ng/mL, 5 ng/mL and 50 ng/mL). No significant interfering compounds, matrix effects or carryover were observed. The validated method provides a sensitive, efficient and robust procedure for the quantification of cannabinoids in blood, using LC-MS-MS-MS3 and a sample volume of 0.1 mL. This work is also a proof of concept for using LC-MS3 technique to determine drugs in biological samples.

Effect of geometry on deformation of anterior implant-supported zirconia frameworks: An in vitro study using digital image correlation.

PURPOSE: To evaluate the effect of geometry on the displacement and the strain distribution of anterior implant-supported zirconia frameworks under static load using the 3D digital image correlation method. METHODS: Two groups (n=5) of 4-unit zirconia frameworks were produced by CAD/CAM for the implant-abutment assembly. Group 1 comprised five straight configuration frameworks and group 2 consisted of five curved configuration frameworks. Specimens were cemented and submitted to static load up to 200N. Displacements were captured with two high-speed photographic cameras and analyzed with video correlation system in three spacial axes U, V, W. Statistical analysis was made using the nonparametric Mann-Whitney test. RESULTS: Up to 150N loads, the vertical displacements (V axis) were statistically higher for curved frameworks (-267.83±23.76µm), when compared to the straight frameworks (-120.73±36.17µm) (p=0.008), as well as anterior displacements in the W transformed axis (589.55±64.51µm vs 224.29±50.38µm for the curved and straight frameworks), respectively (p=0.008). The mean von Mises strains over the surface frameworks were statistically higher for the curved frameworks under any load. CONCLUSION: Within the limitations of this in vitro study, it is possible to conclude that the geometric configuration influences the deformation of 4-unit anterior frameworks under static load. The higher strain distribution and micro-movements of the curved frameworks reflect less rigidity and increased risk of fractures associated to FPDs.