Development and validation of an assay for the measurement of gentamicin concentrations in dried blood spots using UHPLC-MS/MS.

Gentamicin sulfate (GEN) is an aminoglycoside antibiotic with a narrow therapeutic range of plasma concentrations. The collection of venous blood represents a significant burden for patients, especially in neonatology. Dried blood spots (DBS) obtained from capillary blood can be an alternative for drug measurements in this particular population. This study aimed to develop and validate an assay for the quantification of GEN in DBS using UHPLC-MS/MS. Total GEN concentrations were obtained by adding the individual concentrations of the GEN forms C1, C1a, and C2. The assay used a DBS disk containing approximately 17 µL of blood for GEN quantitation in the range of 0.1-40 mg L-1. Measurement accuracy for total GEN was in the range of 102.6-108.6%, inter-assay precision was 11.3-13.1% and intra-assay precision was 9.1-12.8.% GEN was stable for 21 days at - 20 and 8 °C, but only for 24 h at room temperature. Blood Hct affected the accuracy within acceptable limits (93.8-95% at Hct% of 30, 104.3-113% at Hct% of 50). Blood spotted volume did not affect GEN measurement accuracy. Concentrations of GEN in DBS obtained after heel pricks were correlated to plasma levels in a small cohort of neonatal patients. However, percentual differences between estimated plasma concentrations and actual plasma levels presented values between - 64-35.3% (average difference of - 1.9%). The use of DBS for the measurement of GEN concentrations can increase access to TDM of this antibiotic due to the ease of sample collection and the facilitated specimen transportation logistics when testing is not available onsite.

Sensitive determination of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol and complementary cannabinoids in hair using alkaline digestion and mixed-mode solid phase extraction followed by liquid-chromatography-tandem mass spectrometry.

Hair drug testing can be used for the evaluation of cannabis use with a large detection window, and is required for professional driving license granting in Brazil. A positive hair result for cannabis use requires quantification of the metabolite THC-COOH above the cutoff value of 0.2 ng/g. The achievement of such lower limit of quantification is challenging, particularly with the use of liquid chromatography coupled to triple quadrupole mass spectrometers (LC-MS/MS). In this study, a very sensitive LCMS/ MS assay for the simultaneous quantification of THC-COOH along with THC, CBD, and CBN was developed and validated. Sample preparation was based on hair hydrolysis, followed by selective ion-exchange solid-phase extraction. The extraction yield was 101.5-101.6% for THC-COOH, 92.3-97.4% for THC, 89.7-95.2% for CBN, and 104.9-121.1% for CBD. Internal standard corrected matrix effects were - 2.7 to - 1,1 for THCCOOH and - 11.5 to - 0.1% for the other analytes. The lower limit of quantification was 01 ng/g for THC-COOH and 25 ng/g for THC, CBD, and CBN. The assay fulfilled validation guidelines acceptance criteria. The measurement uncertainties were determined and the assay was ISO17025 accredited, being currently used in routine testing.