7  Conclusion

This study presents a comprehensive methodology for extracting SSAO from rat brown adipose tissues, demonstrating effective protein quantification through Bradford and BCA assays, even after defatting with 100% ethanol. Despite storage at -20°C, the samples retained quantifiable SSAO activity, as measured by the Amplex^® Red monoamine oxidase assay. The kinetic parameters obtained for SSAO, with a Km of 0.03193 using benzylamine as a substrate, align closely with previously published results under similar conditions.

However, the Ki values for caffeine and simvastatin were derived from a single experiment, leading to a wider error margin and lower confidence in these results. Additionally, these Ki values fall within the millimolar range, suggesting that both compounds may not be potent inhibitors of SSAO. It is also worth noting that these values were outside the inhibitor concentration range used in their calculation, indicating a need for repeated experiments with adjusted concentrations that reflect the current Ki values. Moreover, the absence of inhibitors with known Ki values as controls limits the ability to accurately compare the inhibition effects of different compounds.

In conclusion, while the study successfully establishes a reliable method for SSAO extraction and activity measurement, further experiments with improved controls and optimized conditions are necessary to yield more reliable Ki values for caffeine and simvastatin. Addressing these limitations in future research will enhance the accuracy and robustness of the findings.