NANOPARTICLE PREPARATION OF SIAM CITRUS PEEL EXTRACT (CITRUS NOBILIS L. VAR. MICROCARPA) USING SHORT-CHAIN CHITOSAN AND TRIPOLYPHOSPHATE AS CROSS LINKER AND CELLULAR UPTAKE STUDY ON MCF-7 CELL LINE BY IN VITRO

Abstract

Objective: High consumption of oranges causes a lot of orange peel waste. Orange peel contains the compound naringenin, which has a cytotoxic effect on various cancer cells. This research aims to develop a preparation of Siamese orange peel extract nanoparticles with short-chain chitosan and tripolyphosphate carriers as an oral drug delivery system and determine its cytotoxic activity against the Michigan Cancer Foundation-7 (MCF-7) cell line. Methods: This research uses the micro tetrazolium (MTT) method to see the cytotoxic activity extract of methanol obtained from maceration extraction. The extract was then formulated into nanoparticles using chitosan and tripolyphosphate. Characterization and evaluation of nanoparticles were carried out, including particle size, zeta potential, entrapment efficiency, and stability in the stomach using 0.1 N HCl and in the intestine using Artificial Intestinal Fluid (AIF) in vitro. This research was also conducted to assess the ability of nanoparticles to enter MCF-7 cells (cellular uptake). Results: Nanoparticles were successfully developed from Siamese orange peel extract. The results of the day 0 nanoparticle characterization were spherical, with average particle size 284.3 nm, zeta potential 0.713 mV, entrapment efficiency 96.73%, and stability in 0.1 N HCl at the 0th hours, respectively. 1st, 2nd, and 3rd. 99.16%, 98.70%, 98.47%, 98.31%, stability on AIF at hours 0, 1, 2, 3 and 4 respectively 99.52%, 99.30%, 99.40%, 98.99%, 99.29%. Characterization of nanoparticles on day 25 showed that the average particle size was 196.2 nm, zeta potential 0.476 mV, entrapment efficiency 96.92%, stability in 0.1 N HCl at 0, 1, 2 and 3 h respectively 99.51%, 98.67%, 98.51%, 98.27%, stability in AIF at 0th, 1st, 2nd, 3rd, and 4th hours 99.24 respectively %, 98.76%, 98.46%, 97.93%, 97.58%. Cytotoxic activity of extract Siamese citrus peel against MCF-7 cells with IC50 of 290.58 µg/ml. The result shows that cellular uptake of Siamese citrus peel nanoparticles can penetrate MCF-7 cells. Conclusion: Stable nanoparticles were successfully developed from Siamese orange peel extract, and their stability was maintained throughout a 30-day storage period. This extract displayed cytotoxic effects and showcased the ability for cellular uptake in MCF-7 cell cultures in vitro.