Development and validation of analytical techniques under good laboratory practices for high complexity preclinical trials

SANTIAGO, G.M.; GÁLVEZ, V.A.; CABAÑA, E; MARELLI, B; HEIN, G; ORTEGA, HH

Mar del Plata

Congreso; Reunión Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2018

Sociedad Argentina de Investigación Clínica

The high prices of biopharmaceutical products demand preclinical assays with small species of animals. Therefore, we aimed to develop and validate analytical micro-methods to determine relevant biomarkers for clinical diagnosis with less than 50 µL of serum sampled from mice, rats and rabbits, in accordance with the principles of Good Laboratory Practices. Serum concentrations of glucose (Glc), triglycerides (Tg), cholesterol (Col), total protein (TP), albumin (Alb) and urea (U) were determined using an ultra-fast UV/Vis spectrometer SPECTROstar Nano (BMG LABTECH GmbH, Germany) and commercial kits designed for human and higher sample volumes. The figures of merit were: limit of detection of 4.71 mg/dL for Glc, 2.72 mg/dL for Tg, 2.72 mg/dL for Col, 0.13 g/dL for TP, 0.05 g/dL for Alb, and 1.25 mg/dL for U ; limit of quantification of 14.29 mg/dL for Glc, 8.25 mg/dL for Tg, 8.25 mg/dL for Col, 0.39 g/dL for TP, 0.16 g/dL for Alb and 3.77 mg/dL for U; quantifiable range of 20-500 mg/dL for Glc, 80-1040 mg/dL for Tg, 50-600 mg/dL for Col, 0.98-9.60 g/dL for TP, 1.55-6.20 g/dL for Alb and 7-140 mg/dL for U. All the techniques were in accordance with international regulations in relation to the intra and inter-precision test, accuracy, inter-laboratories comparison, dilutional linearity and stability at ? 20 °C and ? 80 °C for 60 days. These validated analytical techniques will allow monitoring pathophysiological changes in different animal models, designed for the study of damages occurred in certain organs (such as liver, pancreas, kidney and heart), and involving very small amounts of samples in agreement to the principle of the 3Rs.