PRELIMINARY STUDY ON THE DEVELOPMENT OF UV-VISIBLE SPECTROPHOTOMETRY BASED ON THE ENZYMATIC REACTION OF WHITE RADISH PEROXIDASE (RAPHANUS SATIVUS L.) FOR DRUG ANALYSIS

• Main Author: Anindhia Maharani, Sherin
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/85215
• Institution: Institut Teknologi Bandung

The analysis of drugs has played an important role in quality control and assurance. Spectrophotometry has been a common method for identifying and measuring compounds in an analysis. The presence of chromophores in a compound has been a requirement for analysis using spectrophotometric methods. Derivatization with the peroxidase enzymatic reaction has been a method that can be used to modify compounds without chromophores. One example is the oxidation of phenol groups by hydrogen peroxide (H?O?) in the presence of 4-aminoantipyrine (4-AAP) catalyzed by the enzyme peroxidase, resulting in a quinoneimine that forms a colored product. The aim of this research has been to develop the peroxidase enzymatic reaction for the analysis of drug compounds containing phenol groups. This research has begun with the selection of drug compounds in the Indonesian Pharmacopoeia 6 th Edition that contain phenol groups based on Emerson's criteria, followed by the preparation of peroxidase enzyme solutions from white radish (Raphanus sativus L.), and the study of enzymatic reactions of the selected drug compounds. The results of the selection of drug compounds containing phenols in the Indonesian Pharmacopoeia 6 th Edition have identified 20 drug compounds that meet Emerson's criteria. The isolation of the peroxidase enzyme from white radish (Raphanus sativus L.) using a combination of homogenization, sonication, filtration, centrifugation, and aqueous two-phase system formation methods has resulted in characteristics with a total protein of 4.9 ± 0.01 mg/ml, total activity of 20.9 ± 0.57 U/ml, and specific activity of 4.3 ± 0.11 U/mg. The results of the study of the peroxidase enzymatic reaction from white radish (Raphanus sativus L.) on selected drug compounds have shown a color change from colorless to red with varying red color intensities. Based on all the research stages that have been carried out, this study has shown that the enzyme-based peroxidase reaction has the potential to be developed for drug compounds containing phenols.