Optimisation of liver homogenate procedure to evaluate liver enzymes and lipid biomarkers in mice

Background: Liver disease accounts for 2 million deaths per year worldwide, due to viral hepatitis and hepatocellular carcinoma. Invasive techniques such as liver biopsy remain the gold standard for diagnosing fatty liver diseases and are supported by established scoring systems and indexes. Serum and liver AST and ALT levels are important biochemical indicators of hepatic cellular injury. Analysis of liver enzymes and lipid profiles from serum and liver samples is important in efficacy studies evaluating drugs for liver diseases.Aims and Objectives: This study aimed to optimize liver tissue homogenization by evaluating different media and tissue-to-medium ratios to improve the consistency and reliability of biochemical estimations of liver enzymes and lipid profiles.Materials and Methods: Liver tissues from C57BL/6NCrl mice were homogenized using phosphate-buffered saline (PBS), sucrose, or glycerol at 5%, 10%, and 20% concentrations. Biochemical estimations of ALT, AST, HDL, LDL, total cholesterol (CHOL), and triglycerides (TG) were performed using the Randox Dytona Plus analyzer. Data were statistically analyzed for precision, stability, and reproducibility through relative standard deviation (RSD) and mean variability.Results: Among all tested conditions, the 20% homogenate in PBS demonstrated consistent and reproducible results, yielding enzyme activities and measurable lipid markers within or near the instrument’s detectable range. Precision analysis showed low %RSD values (