Alternative methods for endotoxin evaluation: international challenges and trends for patient safety

• Grant number: 18/19237-4
• Support Opportunities: Regular Research Grants
• Start date: February 01, 2019
• End date: September 30, 2021
• Field of knowledge: Health Sciences - Pharmacy - Medicines Analysis and Control
• Principal Investigator: Terezinha de Jesus Andreoli Pinto
• Grantee: Terezinha de Jesus Andreoli Pinto
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Adriana Aparecida Buzzo Almodovar ; Adriana Bugno ; Carla Lilian de Agostini Utescher ; Daniela Dal Molim Ghisleni ; Marco Aurélio Floriano Piantola ; Rui Curi ; Viviane Fongaro Botosso ; Wagner Quintilio

Abstract

Even nowadays endotoxins are a major concern, especially in regard to parenteral drugs due to their ability to cause intense biological effects, in both humans and animals. The pyrogen test (in vivo) is widely used in the analysis of parenteral drugs with the aim of releasing the finished product for consumption. However, it makes use of rabbits, which is an extremely controversial issue due to 3Rs principles (substitution, reduction and refinement), a global trend that defend the reduction of animal testing and the development of in vitro tests to replace them. Animal-derived Limulus amebocyte lysate (LAL) reagent has been widely used for the detection of bacterial endotoxins and could replace the pyrogen test, but it may present problems in regard to the low recovery of endotoxin, as well as interference from certain components leading to a false-positive or a false- negative result, strongly compromising the safety of patients using this medicine. It is importante to underscore that crab population has been in decline, which emphasizes the importance of new sensible and effective alternative methods, perfectly applicable in routine laboratory. Based on what was described above, the main goals of the project were the development and validation of an alternative method to evaluate bacterial endotoxins in small volume parenteral drugs (serum hyperimmune ), as well as large volume parenteral drugs (0.9% saline solution) by an end-point florescence-based endotoxin method using the rFC reagent which aimed, respectively, animal testing reduction in quality control trials, and the availability of an alternative method for the detection of small amounts of endotoxins, promoting the safety of patients in general. (AU)