doi.org/10.36721/PJPS.2020.33.2.REG.505-510.1 Pak. J. Pharm. Sci., Vol.33, No.2, March 2020, pp.505-510 505 Terpenoid-lupeol of red dragon fruit ( Hylocereus polyrhizus) and its immunomodulatory activity Sri Wahdaningsih 1,2 , Subagus Wahyuono 3 , Sugeng Riyanto 3 and Retno Murwanti 3 1 Departement of Pharmacy, Faculty of Medicine, Tanjungpura University, Pontianak, Indonesia 2 Graduate Program, Faculty of Pharmacy, Gadjahmada University, Yogyakarta, Indonesia 3 Faculty of Pharmacy, Gadjahmada University, Yogyakarta, Indonesia Abstract: Red dragon fruit (Hylocereus polyrhizus, (F.A.C. Weber) Britton and Rose) has been reported to have various biological activities such as antimicrobial, anti-hypercholesterolemia, anti-diabetes mellitus, cardiovascular risk reduction, health supplement, and melanoma cell inhibitory. The red thick peel of this fruit is just practically a waste that is possibly utilized to maintain health, therefore this research aimed to isolate and identify active compounds of H. Polyrhizus peels which can improve the immune system of body. In order to simplify methanol extract was partition and fractionation. The active compounds of petroleum ether fraction were separated and purified using preparative thin layer chromatography. The identification of the compounds structure was conducted through spectroscopic techniques, including UV, FT-IR, 13 CNMR and 1 HNMR spectroscopy. The data of spectra revealed that the isolate is lupeol. The statistical analysis of macrophage activity showed that the isolate with concentrations of 100, 50, 25, 12.5 and 6.25µg/mL could activate the macrophages higher than control negative. Terpenoid generated from the isolation of Hylocereus polyrhizus was identified as lupeol (1-isopropenyl-3a,5a,5b,8,8,11a-hexamethyl-eicosahydrocyclopenya [α] chrysen-9ol. In vitro test shows that the isolated compound had an immunomodulatory activity by increases macrophage phagocytosis of latex beads. Keywords: H. polyrhizus, petroleum ether fraction, terpenoids - lupeol, immunomodulator. INTRODUCTION Dragon fruit is cactus subspecies of the genus Hylocereus of the family Cactaceae. The genus has about 16 species; one of them is red dragon fruit. The peel of red dragon fruit is a rich source of natural compounds such as phenolics, flavonoids, carotenoids, anthocyanins and terpenoids. It also contains pectin, galacturonic acid, mannose, and galactose, xylose and rhamnose (Muhammad et al., 2014). The immune system needs antioxidant nutrients for hematopoiesis, cell membrane protection against Reactive oxygen species (ROS) and elimination pathogenic microorganisms. The diseases caused by Reactive oxygen species (ROS) include rhematoid arthritis, hemorrhagic shock, cardiovascular, metabolic disorders, neurodegenerative diseases, gastrointestinal, AIDS, etc (Aher et al., 2011). Immunomodulator is a substance that affects the immune system. The immunity consists of innate-immune system and adaptive immune system with a very complex biochemical systems. Some compounds which can be immunodulator are polysaccharides, terpenoids, saponins, alkaloids, isoflavonoids, glucosides, tannins, fatty acids, steroids, triterpenes, and flavonoids (Wagner et al., 1991; Parmar et al., 1997; Roshan and Savitri, 2013). This research aimed to isolate and identify active compounds of H. Polyrhizus peels which can improve the immune system of body. MATERIALS AND METHODS Plant materials: The sample used in this research was the pericarp of red dragon fruit obtained from Bantul, Yogyakarta, Indonesia. The total amount of the fruit used was 30.29 kg. Of the amount, the peel generated was 6.79 kg or 23.683% of the total fruit weight. The obtained simplisia was 800 grams. Extraction, fractionation, isolation Maceration was performed by dissolving 800 grams of simplisia in methanol at room temperature. The generated methanol extracts were partitioned with petroleum ether (PE) solvent. The soluble fractions of petroleum ether were fractionated with vacuum liquid chromatography (VLC). About 2.0 grams of the fractions were fractionated. The static phase used was silica gel 60 for column chromatography with increasing polarity gradient of mobile phase. Petroleum ether: ethyl acetate [50 ml PE], [49:1], [47.5:2.5], [45:5], [42.5:7.5], [40:10], [37.5:12.5], [35:15], [30:20] and [25:25] were used as the mobile phase. The eluates of each solvent comparison were colected. It generated 10 fractions which were then analyzed by thin-layer chromatography with promoter petroleum ether: Ethyl acetate [10:1] and spayed with *Corresponding author: e-mail: wahdanieanie@gmail.com