Indonesian Journal of Chemical Research http://ojs3.unpatti.ac.id/index.php/ijcr Indo. J. Chem. Res., 8(2), 15-20, 2020 DOI: 10.30598//ijcr. 2020.8-yun 15 Stability Indicating RP-HPLC for Quantification Mangiferin in Extract of Three Species Mango Leaves Yuni Retnaningtyas*, Nia Kristiningrum, Hidayah Dwi Renggani, Indah Purnama Sary 1 Departement of Chemistry, Faculty of Pharmacy, University of Jember Jl. Kalimantan 37 Kampus Tegal Boto Jember * Corresponding Author: ifir_retnaningtyas@yahoo.co.id Received: 2019-12-20 Received in revised: 2020-4-29 Accepted: 2020-5-28 Available online:2020-5-31 Abstract The stability indication of Reversed Phase-High Performance Liquid Chromatography (RP-HPLC) method was validated for quantitative determination of mangiferin on three species mango leaves (Mangifera odorata Griff, Mangifera foetida Lour, and Mangifera indica L.). The samples were extracted by maseration method using methanol and concentrated using rotary evaporator. The method carried out on stationary phase a purospher RP-18 endcapped (25 cm × 4.6 mm i.d., 5 μm) column with a mobile phase consisting of methanol: phosphoric acid 0.1% (v/v) (31:69); flow rate:0.8 mL/min; solvent methanol, detection was carried out at 258 nm. The analytical performace this measurement is good with the value of linearity (r 2 =0.998), precision (%RSD=0.649%), and accuration (10.67%). The forced degradation studies were carried out according to the International Conference on Harmonization (ICH) guidelines. The results indicating that the complete separation between degradation products and mangiferin peak occured. The degradation limit of mangiferin 5–20% (according to the guideline of ICH) except in basic condition (100%). The method was succesful applied to determine of the mangiferin in pakel (Mangifera foetida), kweni (Mangifera indica) and kopyor (Mangifera odorata) extract. The mangiferin content was obtained are pakel (9.95%), kopyor (7.40%) and kweni ( Mangifera odorata) (2.49%) respectively. Keywords: Mangiferin, Mango leaf, Mangifera odorata Griff., Mangifera foetida Lour, Mangifera indica L., RP-HPLC, validation INTRODUCTION The Mango was easily found in Indonesia and distributed in all region of Indonesia. Indonesia was one of the country with the highest production of mango (Husen et al., 2012). But, until now only the fruit have been used while the leaves not yet. Mango leaves (Mangifera indica L.) from Anarcadiaceae family contains many chemical compounds such as phenol, β-carotene, flavonoid, tannin, saponin, alkaloid and steroid (Palafox-Carlos et al., 2012 ; Pino et al., 2011). One of the phenolic compounds that found in mango was mangiferin. Mangiferin can be found in all parts of mango plants as peel, pulp, seed kernel (Luo et al., 2012) , bark (García-rivera et al., 2011) and leaves (Jutiviboonsuk et al., 2010). The mangiferin was a phenolic compound that has poten antioxidant activity, and multifactorial pharmacological effects, including antidiabetic, antitumor, lipometabolism regulating, cardioprotective, anti-hyperuricemic, neuroprotective, anti-inflammatory, antipyretic, analgesic, antibacterial, antiviral and immunomodulatory effects (Mirza et al., 2013; Du et al., 2018). Mangiferin is c- glucosyl xanthone and its structures (Figure 1) 2-C-D- glucopyranosyl-1,3,6,7-tetrahydroxy xanthone, fulfill to Lipinski rules (Campa et al., 2012). Based on chemotaxonomy, mangiferin can be found in another mango species; more related the plant so the chemical compound will be more similar (Subha et al., 2007). But, the amount of the compound can be affected by some factors such as the location of cultivation, variety, and stage of maturity. Figure 1. Structure of Mangiferin Mangiferin was quantified only in one species of mango (Mangifera indica L.) while there are 62 species of mango (Pracaya, 2005). Quantification of mangiferin can be done using High-performance thin layer chromatography (HPTLC) (Subha, et al., 2007),