VALIDATION OF BIOCHEMICAL AND HEMATOLOGICAL PARAMETERS USED TO MONITOR PRECLINICAL EXPERIMENTAL DIABETES MELLITUS IN DIFFERENT SPECIES.

B Moreno-Lobato, FM Sánchez-Margallo, D Celdrán, MF Martín-Cancho. Minimal Invasive Surgery Centre Jes ú s Us ó n, Pharmacy Unit-10071, Spain

Introduction

The increased incidence of diabetes is one of the bases for the growing interest in pharmacological research about the treatment of this disease1. In this work, the use of experimental animal models is essential for the search of new therapeutic tools 2. The quality of these studies, which usually require the induction of experimental diabetes, can be ensured by the use of Good Laboratory Practices (GLPs). Clinical follow-up of experimental studies in animals requires biochemical and hematological tests that must be validated 3, 4. Thus, through a validation process, it can be verified if the method is reliable enough and whether the results are obtained within the preset conditions 5.

The objective of this study is to verify and document the validation of the analytical methods of the parameters analyzed during the clinical follow-up of diabetic animals, with the objective of ensuring the quality and integrity of the data obtained at the clinical analysis laboratory in order to finally ensure the quality of the results obtained about the efficacy and activity of new therapies or therapeutic approaches tested in preclinical studies.

Material and Methods

Three animal species, commonly used in the study of diabetes mellitus, were selected for this study: pig, dog and rabbit. After receiving the blood samples at the laboratory, hematology analysis were performed (Nihon Kohden Iberica S.L., Madrid, Spain) and in order to evaluate the metabolism glucose, cholesterol and triglycerides determinations were performed (Metrolab 2300, UV-Vis Metrolab SA. Buenos Aires, Argentina).

The validation process was carried out by performing a repeatability study of the method (6 repetitions of the same sample from each animal by the same analyst) and also by calculating the intermediate precision (2 repetitions carried out by 3 different analysts). With the data obtained in each species a descriptive statistical analysis was performed, calculating the mean, standard deviation and coefficient of variation for each analyzed parameter.

Results

Coefficient of variation values below 3.5% were obtained in this study for all biochemical parameters analyzed. In the hematology study, the values obtained were below 4% for all parameters in the different animal species used, except for platelets study in rabbits, where coefficients of variation obtained were 5.86% for the repeatability study and 7.91% for the intermediate precision.

Conclusion

The purpose of method validation is to ensure the outcome of high quality data, ensuring good and reproducible results. The results of the validation study performed show excellent coefficients of variation, it can be concluded that the analytical methods used are repetitive, and thus can be used for routine measurement of animal samples from the studied species providing accuracy in diabetes preclinical studies.

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