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Evaluation of Liver Care Function

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With over 15 years' experience in providing preclinical testing and research services for the veterinary supplement industry, CD Formulation has helped many companies advance formulations that support liver health in dogs and cats. Through biomarkers of hepatotoxicity, assessments of metabolic processes, and studies examining supplement impacts over time, we can generate robust data on a product's potential to maintain or improve liver health.

Liver Disease Models We Can Build

Carbon tetrachloride-induced chemical liver injury.
Chemical hepatitis injury induced by the analgesic acetaminophen.
Chronic liver injury induced by thioacetamide-induced liver fibrosis.
Alcoholic fatty liver induced by alcoholic liquid feed.
Non-alcoholic fatty liver disease induced by high-fat diet.

What Can We Do for Liver Protection Function Evaluation?

Glutathione (GSH) Concentration Analysis	Liver tissue samples are collected from test animals and homogenized. The homogenate is deproteinized to remove interfering proteins. Total glutathione, both reduced (GSH) and oxidized (GSSG), is derivatized to form a fluorescent adduct quantified by HPLC with fluorescence detection.
Glutathione Peroxidase (GPx) Activity Analysis	We prepare liver homogenates from tissue samples. The reaction mixture contains glutathione, glutathione reductase, NADPH, and hydrogen peroxide as a substrate. Glutathione peroxidase in the homogenate catalyzes the reduction of hydrogen peroxide, oxidizing glutathione. The rate of NADPH oxidation, which is proportional to glutathione peroxidase activity, is measured spectrophotometrically.
Glutathione Reductase (GRd) Activity Analysis	Glutathione reductase catalyzes the reduction of oxidized glutathione (GSSG) back to reduced glutathione (GSH). Liver homogenates are prepared from tissue samples. The reaction mixture contains GSSG, NADPH, and glutathione reductase from the homogenate. Glutathione reductase converts GSSG to GSH, oxidizing NADPH in the process. We detect the decrease in NADPH absorbance spectrophotometrically over time, which is directly proportional to glutathione reductase activity levels.
Superoxide Dismutase (SOD) Activity Analysis	The assay mixture contains xanthine, xanthine oxidase (a superoxide generator), nitroblue tetrazolium dye, and SOD from the homogenate sample. Xanthine oxidase generates superoxide anions which reduce nitroblue tetrazolium, producing a colored formazan product. SOD in the sample prevents this reaction by catalyzing the dismutation of superoxide to hydrogen peroxide and oxygen. The decrease in formazan formation due to the presence of SOD is quantified spectrophotometrically.
Catalase (CAT) Activity Assay	The reaction mixture contains hydrogen peroxide substrates and catalase from the homogenates. Catalase catalyzes the decomposition of hydrogen peroxide. The reaction is monitored spectrophotometrically by measuring the decrease in the absorbance of hydrogen peroxide over time.
Histopathological Section Observation	Liver tissue samples are collected and fixed in neutral buffered formalin. The samples are then dehydrated through a series of graded alcohols, cleared with xylene, and infiltrated and embedded in paraffin wax. Sections of liver tissue are cut using a microtome and mounted on glass slides. The slides are stained, most commonly with hematoxylin and eosin (H&E), which facilitates cellular-level examination. A pathologist then examines the stained slides under a light microscope. They evaluate the hepatic architecture and cellular structure, looking for any abnormal pathological changes compared to controls. Features assessed include signs of inflammation, necrosis, fibrosis, fatty changes, and morphological integrity.
Other Projects	We can also record the weekly body weight, liver weight/body weight (%), mortality and other related measurement items of animals during chronic experiments.

Why Choose Our Liver Care Function Evaluation Services

Comprehensive assessment of hepatic function through validated clinical, biochemical, histological and molecular testing.
Generate data on impacts on antioxidant defenses, detoxification capacity, and ability to mitigate oxidative stress and toxic insults.
Customized studies designed to demonstrate efficacy for liver support, protection from toxicity, or improving existing conditions.
Insights into potential mechanisms of action through specialized techniques like metabolomics and gene expression analysis.

CD Formulation provides a thorough preclinical assessment of liver function. Through clinical chemistry, histopathology, and multi-omics analysis, we can generate robust data on liver health. Contact us today to discuss your research goals!

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