<![CDATA[Rhizophora apiculata fruit potentially contains carotenoid pigments that may contribute to the biological properties of mangrove plants. This study aimed to separate and preliminarily identify carotenoid pigments from R. apiculata fruit and to predict the molecular interaction of zeaxanthin with inducible nitric oxide synthase (iNOS). Pigments were extracted with ethanol, partitioned into an n-hexane phase, and separated by thin-layer chromatography on silica gel using acetone-petroleum ether (70:30) as the mobile phase. Pigments were characterized based on zone color, retardation factor (Rf), and UV-Vis absorption spectra. Molecular docking was performed against the human iNOS oxygenase domain (PDB ID: 3E7G) using Molegro Virtual Docker 6.0, with AT2 as the co-crystallized ligand for redocking validation. TLC separation produced seven zones with Rf values ranging from 0.62 to 1.00. TLC-derived fraction F1 was orange and exhibited absorption maxima at 425, 453, and 480 nm, supporting its preliminary identification as β-carotene. Fraction F3 was yellow, had an Rf value of 0.83, and showed a maximum absorption at 475 nm, supporting its preliminary identification as zeaxanthin. Redocking validation produced a mean RMSD of 0.9223 ± 0.0372 Å. Zeaxanthin yielded mean MolDock and Rerank Scores of −153.419 ± 19.994 and −120.843 ± 20.801, respectively, which were more negative than those obtained for AT2. Zeaxanthin interacted with several residues also recognized by AT2, including Trp372, Gly371, Trp194, Cys200, Met374, and Met434, with predominantly hydrophobic interactions. These findings provide preliminary evidence for the presence of β-carotene and zeaxanthin in R. apiculata fruit and support the predicted interaction of zeaxanthin within the iNOS binding pocket. Keywords: Rhizophora apiculata; carotenoids; thin-layer chromatography; iNOS; molecular docking ABSTRAK Buah Rhizophora apiculata berpotensi mengandung pigmen karotenoid yang berkontribusi terhadap aktivitas biologis tumbuhan mangrove. Penelitian ini bertujuan memisahkan dan melakukan identifikasi awal pigmen karotenoid pada buah R. apiculata serta memprediksi interaksi molekuler zeaxanthin terhadap inducible nitric oxide synthase (iNOS). Pigmen diekstraksi menggunakan etanol dan dipartisi ke dalam fase n-heksana, kemudian dipisahkan menggunakan kromatografi lapis tipis pada pelat silika gel dengan fase gerak petroleum eter dan aseton (70:30). Karakterisasi pigmen dilakukan berdasarkan warna zona, nilai retardation factor (Rf), dan spektrum serapan UV-Vis. Penambatan molekuler dilakukan terhadap domain oksigenase iNOS manusia (PDB ID: 3E7G) menggunakan Molegro Virtual Docker 6.0, dengan AT2 sebagai ligan ko-kristal untuk validasi redocking. Pemisahan KLT menghasilkan tujuh zona dengan nilai Rf 0,62-1,00. Fraksi hasil KLT F1 berwarna oranye dengan serapan maksimum pada 425, 453, dan 480 nm sehingga diidentifikasi secara awal sebagai β-karoten, sedangkan F3 berwarna kuning dengan 0,83 dan serapan maksimum pada 475 nm sehingga diidentifikasi secara awal sebagai zeaxanthin. Validasi redocking menghasilkan RMSD rata-rata 0,9223 ± 0,0372 Å. Zeaxanthin memberikan MolDock Score dan Rerank Score rata-rata masing-masing sebesar −153,419 ± 19,994 dan −120,843 ± 20,801, yang lebih negatif dibandingkan AT2. Zeaxanthin berinteraksi dengan beberapa residu yang juga dikenali AT2, yaitu Trp372, Gly371, Trp194, Cys200, Met374, dan Met434, dengan pola interaksi yang didominasi kontak hidrofobik. Hasil penelitian menunjukkan keberadaan awal β-karoten dan zeaxanthin pada buah R. apiculata serta mendukung kemungkinan interaksi zeaxanthin pada kantong pengikatan iNOS. Kata kunci: Rhizophora apiculata; karotenoid; kromatografi lapis tipis; iNOS; penambatan molekuler]]>
