Due to bacteria being unable to metabolize molecules, the mutagenic potential of samples is assessed directly and in the presence of metabolic activation system: Microsomal fraction S9 of Phenobarbital/ß-Naphtoflavone induced rat liver homogenate. Rat liver S9 is used in the Ames Test (OECD 471), in vitro Micronucleus Test (MNvit, OECD 487) and in Chromosomal Aberration Test (OECD 473) for chemicals, xenobiotics, herbicides, pesticides, biocides, cosmetics, pharmaceutical preparations or soil, waste water.
Phenobarbital/ß-Naphtoflavone induced hepatic S9 after reconstitution is a buffered solution containing the post mitochondrial S9 fraction of a pool of livers from male SpragueDawley rats. Liver S9 fractions (the 9000g supernatant of a liver homogenate) contain both microsomal and cytosolic fractions. S9 fraction includes many cytosolic enzymes: N-acetyl transferases, methyltransferases, sulfotransferases, glutathione transferases, aldehyde oxidase, xanthine oxidase. Both Phase I (Cytochrome P450-Monooxigenases) and Phase II metabolites act simultaneously on test samples. Due to the fact that the enzymes in the S9 fractions are diluted significantly, cofactors such as e.g. β-Nicotinamide adenine dinucleotide phosphate-regenerating system (NADPH, Phase I oxidation) are required for activity. Cofactors are added to the S9 preparation before the experiment (S9 Cofactor Kit, art.no. PCO-0800 or PCO-1800).
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