Y. Susilawati et al. Int J App Pharm, Vol 13, Special Issue 4, 2021, 234-238 1 st Bandung International Teleconference on Pharmacy (BITP), 2021 | 234 ISOLATION AND DETERMINATION CARPAINE ALKALOID IN PAPAYA (CARICA PAPAYA L.) LEAF EXTRACT BY THIN-LAYER CHROMATOGRAPHY SCANNER Original Article YASMIWAR SUSILAWATI 1,2* , NYI MEKAR SAPTARINI 3 , ZELIKA MEGA RAMADHANIA 2 , ELI HALIMAH 4 , NAILAH NURJIHAN ULFAH 2 1 Herbal Study Center, Faculty of Pharmacy, Padjadjaran University, Jalan Raya Bandung-Sumedang KM 21, Jatinangor, West Java, Indonesia, 2 Department of Biology Pharmacy, Faculty of Pharmacy, Padjadjaran University, Jalan Raya Bandung-Sumedang KM 21, Jatinangor, West Java, Indonesia, 3 Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Padjadjaran University, Jalan Raya Bandung-Sumedang KM 21, Jatinangor, West Java, Indonesia, 4 Department of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Padjadjaran University, Jalan Raya Bandung-Sumedang KM 21, Jatinangor, West Java, Indonesia * Email: yasmiwar@unpad.ac.id Received: 20 Aug 2021, Revised and Accepted: 02 Oct 2021 ABSTRACT Objective: This research was conducted to isolate the alkaloid carpaine by chromatography method and to determine it quantitatively by Thin Layer Chromatography Scanner. Methods: Dried leaves were macerated with ethanol 70% and fractionated with dichloromethane. Isolation of carpaine alkaloid from the dichloromethane fraction was carried out by column chromatography and preparative thin-layer chromatography according to the Rf value in Thin Layer Chromatography (TLC) after exposure by Dragendorff reagent. Results: The content of carpaine alkaloid was 7.5 mg with Rf 0.58 and dichloromethane: methanol (9.2:0.8) as eluent. Validation showed the linearity (R 2 ) 0.9988, the limit of detection(LOD) was 0.05 ppm, the Limit of Quantification (LOQ) was 0.19 ppm, the recovery from 98.93-102.43%, and the % coefficient of variation was 0.16%. Conclusion: Carpaine alkaloid in papaya leaf extract was 10.52%. Keywords: Carica papaya, Papaya leaf, Carpaine alkaloid, Validation method, TLC scanner © 2021 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2021.v13s4.43867 Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION Papaya (Carica papaya L.) comes from the family Caricaceae. This plant is cultivated in almost all subtropical and tropical countries such as South America, India, Phillippines and Indonesia. Almost all parts of the papaya plant have medicinal properties [1]. Papaya leaves extract have properties as hematopoietic, antimalarial, antifungal, anti-HIV-1, hypoglycemic, anti-inflammatory, antimicrobial, anti-tumor, immunomodulatory, hepatoprotector, and anti-thrombocytopenic activity [2-10]. In addition, papaya leaves can significantly increase platelet counts and increase the number of red blood cells [11] and showed analgesic activity in mice compared to aspirin [12]. Papaya leaves contain alkaloids carpaine, dehydrocarpaine I and II, pseudocarpaine, choline, carposide, flavonols, benzylglucocinolate, papain, tannins, vitamin C, and E [13]. The major component of papaya leaves is carpaine alkaloid. This compound reported can reduce blood pressure, antitumor and antiplasmodial activity [14]. In relation to lowering blood pressure, it was reported that carpaine showed a direct effect on the myocardium, where this effect was thought to be related to the macrocyclic dilactone structure, a possible cation chelating structure [15]. The isolated carpaine is able to maintain mouse platelets up to 555±85.17 x 109/L which has been induced by busulfan for 20 d with a dose of 200 mg/kg papaya leaf extract containing 2 mg/kg carpaine alkaloid [16]. Fig. 1: Carpaine structure [17] Carpaine alkaloid found with molecular docking has the potential to be antiviral dengue because they have interactions with Thr413 amino acid residues against RNA-dependent RNA polymerization in dengue viruses [18]. The research of Wang et al. (2015) states that the levels of carpaine alkaloid in papaya leaves are 0.93 g/kg by High- Performance Liquid Chromatography (HPLC) method [19]. Chromatography is a simple method that can be used to identify secondary metabolites in plants, including alkaloids [20]. Densitometry method with TLC scanner has been used at research of alkaloid compounds in Chelidonium majus L. plants which are capable of detecting many samples at the same time [21]. The densitometry method is simpler than HPLC in determining the level of identity compounds in an extract. This method has the advantages like being more effective in preparation and cost-efficient, even though it has a less sensitive sensitivity compared to HPLC. Determination of levels in extracts with TLC scanner is needed validation to determine a method that can use as the determination requirements. Therefore, this study aimed to isolate and determine carpaine alkaloids from papaya leaf extract Carica papaya L. using the TLC densitometry method. MATERIALS AND METHODS Plant material Papaya (Carica papaya L.) leaves taken from botanical garden Manoko Lembang, Bandung, West Java, Indonesia. The plant identification was determined in the Taxonomy Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Indonesia. Chemical materials All solvents and chemicals used to prepare Dragendorff, Mayer, and Liebermann-Burchard reagents had an analytical grade purchased from Merck, including silica gel GF254. International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 13, Special Issue 4, 2021