PROFIL METABOLIT BERBAGAI EKSTRAK DAUN Chrysophyllum cainito L. MENGGUNAKAN UPLC-QTOF-MS/MS

ABSTRACT Chrysophyllum cainito L. is a plant which empirically used as traditional medicine. The pharmacological effect of C. cainito is caused by secondary metabolite activity contain in the leaves. The aim of this research was to know the metabolites profile in n-hexane extract, ethyl acetate extract, and methanol extract of C. cainito leaves using UPLC-QToF-MS/MS. Dried powder of C. cainito leaves was extracted with n-hexane, ethyl acetate, and methanol with gradual extraction using Ultrasonic Assisted Extraction (UAE). Each extract was prepared with methanol and DCM solvent then injected 5 µl into UPLC-QToF-MS/MS and analyzed with Masslynx 4.1 softwares and Chemspider. The result showed that there were 28 compounds from n-hexane extract with diethyltoluamide as major compound, 47 compounds from ethyl acetate extract with loliolide as major compound, and 34 compounds from methanol extract with eplerenone as major compound. Based on literature study, there were also several compounds that likely having activity as phytoestrogens. Keyword: Metabolite profiling, chrysophyllum cainito L., UPLC QToF-MS/MS     ABSTRAK Chrysophyllum cainito L. merupakan tumbuhan yang secara empiris digunakan sebagai obat tradisional. Efek farmakologi tersebut disebabkan adanya aktivitas dari berbagai senyawa metabolit sekunder yang terkandung dalam daun C. cainito. Tujuan dari penelitian ini untuk mengetahui profil metabolit ekstrak n-heksana, ekstrak etil asetat, dan ekstrak metanol daun C. cainito menggunakan UPLC-QToF-MS/MS. Serbuk kering daun C. cainito diekstraksi secara bertingkat menggunakan n-heksana, etil asetat, dan metanol dengan metode UAE. Masing-masing ekstrak dipreparasi dengan metanol dan DCM lalu diinjeksikan sebanyak 5 µl ke dalam UPLC-QToF-MS/MS, kemudian dianalisis dengan software Masslynx 4.1 dan Chemspider. Hasil menunjukkan profil metabolit dari masing-masing ekstrak daun C. cainito, yaitu ekstrak n-heksana dengan 28 senyawa dan diethyltoluamide sebagai senyawa mayor, 47 senyawa terkandung dalam ekstrak etil asetat dengan senyawa mayor loliolide, dan 34 senyawa terkandung dalam ekstrak metanol dengan senyawa mayor yaitu eplerenone. Dari studi literatur diketahui terdapat beberapa senyawa yang memiliki aktivitas sebagai fitoestrogen. Kata Kunci: Metabolite profiling, Chrysophyllum cainito L., UPLC QToF-MS/M

Uji toksisitas ekstrak akuades (suhu kamar) dan akuades panas (70 oC) daun kelor (Moringa oleifera Lamk.) terhadap larva udang Artemia salina Leach

Kelor (Moringa oleifera Lamk.) adalah tanaman yang kaya nutrisi. Kandungan nutrisi tersebar pada seluruh bagian tanaman kelor, mulai dari daun, kulit batang, bunga, buah (polong), sampai akarnya. Kelor (Moringa oleifera Lamk.) mempunyai banyak manfaat, misalnya digunakan sebagai koagulan, nutrisi, vitamin, dan sebagai obat. Selama ini kelor di Indonesia hanya digunakan sebagai tanaman pagar dan sayuran dan masih jarang ada penelitian tentang bioaktivitas daun kelor dan pemanfaatannya sebagai antikanker. Penelitian ini mempelajari toksisitas ekstrak akuades (suhu kamar) dan akuades panas (70 oC) daun kelor (Moringa oleifera Lamk.) terhadap larva udang Artemia salina Leach dengan menggunakan metode BSLT. Pemilihan pelarut akuades sangat menguntungkan karena ekonomis dan mudah diperoleh, sehingga mudah diaplikasikan oleh masyarakat.Hasil penelitian menunjukkan bahwa ekstrak akuades (suhu kamar) dan akuades panas (70 oC) daun kelor memiliki tingkat toksisitas terhadap Artemia salina Leach yang ditunjukkan dengan nilai LC50 kurang dari 1000 ppm. Ekstrak akuades panas (70 oC) memiliki toksisitas lebih baik dari pada ekstrak akuades (suhu kamar) karena dihasilkan nilai LC50 berturut-turut 163,979 ppm dan 265,977 ppm. Kandungan golongan senyawa pada ekstrak akuades panas (70 oC) adalah alkaloid, flavonoid, tanin dan triterpenoid. Berdasarkan hasil tersebut bahwa daun kelor mempunyai potensi sebagai tanaman sediaan herbal yang nantinya bisa dimanfaatkan sebagai antibakteri dan antikanker

Antiviral Activitiy of Curcumin, Demethoxycurcumin, Bisdemethoxycurcumin and Cyclocurcumin compounds of Curcuma longa against NSP3 on SARS-CoV-2

SARS-CoV-2 genome encodes two large polyproteins (pp), pp1a and pp1ab which are cleaved and transformed into a mature form by a protease, non-structural protein 3 (NSP3). NSP3 is encoded by open reading frame (ORF) 1a/b. Curcuma longa (C. longa) or turmeric has been documented to have antiviral effects. The aim of this study was to assess the viral activities of C. longa against SARS-CoV-2 focusing on its potency to inhibit viral replication by targeting NSP3. PubChem databases were used to obtain the metabolic profile of C. longa. The compound's interaction with nucleocapsid was analyzed using molecular docking with Molegro Virtual Docker. Bioinformatics analysis based on rerank score presents all compounds of C. longa have higher binding affinity than the native ligand with cyclocurcumin as the lowest score (-128.38 kcal/mol). This anti-viral activity was hypothesized from the similarity of hydrogen bonds with amino acid residues Ser 128 and Asn 40 as key residues present in Ribavirin. This study reveals that C. longa is the potential to be developed as an antiviral agent through replication inhibition in SARS-CoV-2 targeting its replication mediated by NSP3.Keywords: C. longa, Non-Structural Protein 3, COVID-19

Potential Inhibition of Melaleuca leucadendron L. Compounds Against the NSP5 SARS CoV-2 Protein

COVID-19 is an infectious disease caused by Severe Acute Respiratory Syndrome (SARS-CoV-2), causing a global health emergency as a pandemic disease. The lack of certain drug molecules or treatment strategies to fight this disease makes it worse. Therefore, effective drug molecules are needed to fight COVID-19. Non Structural Protein (NSP5) or called Main Protease (Mpro) of SARS CoV 2, a key component of this viral replication, is considered a key target for anti-COVID-19 drug development. The purpose of this study is to determine whether the compounds in the Melaleuca leucadendron L. plant such as 1,8-cineole, terpene, guaiol, linalol, α-selinenol, β-eudesmol and γ-eudesmol are predicted to have antiviral activity for COVID-19. Interaction of compounds with NSP5 with PDB code 6WNP analyzed using molecular docking with Molegro Virtual Docker. Based on binding affinity, the highest potential as an anti-viral is Terpineol with binding energy (-119.743 kcal/mol). The results of the interaction showed that terpinol has similarities in all three amino acid residues namely Cys 145, Gly 143, and Glu 166 with remdesivir and native ligand. Melaleuca leucadendron L. may represent a potential herbal treatment to act as: COVID-19 NSP5, however these findings must be validated in vitro and in vivo.Keywords: COVID-19, In Silico, NSP5/ 6WNP, Melaleuca leucadendron L

ANTIVIRAL ACTIVITY OF SAUSSUREA LAPPA ETHANOL EXTRACT AGAINST SARS-COV-2: IN VITRO STUDY

Objective: The study aims to investigate the antiviral activity of S. lappa against Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2) in vitro. Methods: The extracts are obtained by ultrasonic-assisted extraction (UAE) with a 96% ethanol solvent. Thin-layer chromatography (TLC) uses n-hexane: ethyl acetate and chloroform: methanol) as mobile phases. The staining outcome is subsequently examined using UV visualizers with a wavelength of 366 nm. To assess the antiviral activity of Vero E6 cells, extracts were employed at doses of 25, 50, 75, and 100 µg/ml, with remdesivir serving as the positive control. Supernatants were collected on days 1, 2, 3, and 6 for qRT-PCR testing with target genes E and ORF1ab. Time-addition experiments were conducted to determine how the extract works as antiviral. Protein expression was tested with Western blots with antibodies S and N SARS-CoV-2. Results: TLC identifies terpenoid chemicals present in the ethanol extract of S. lappa. The ethanol extract of S. lappa exhibited antiviral effects against SARS-CoV-2, with an inhibitory concentration 50 (IC50) of 40 µg/ml, a cytotoxic concentration 50 (CC50) of 131.4 µg/ml, and a selectivity index of 3.51. The extract can potentially impact the entry-post-entry phase of SARS-CoV-2 infection in Vero cells. The immunoblotting results demonstrated a reduction in the expression of S and N proteins in the treatment group compared to the negative control. Conclusion: S. lappa ethanol extract has antiviral activity against SARS-CoV-2 based on an in vitro study