Bioactive Composition and Antioxidant Characteristics of Calophyllum inophyllum Seed Flour: Potential Relevance for Functional Food Development

Azizul Berlyansah; Andreanyta Meliala; Paramita Narwidina; Wira Mondana; Beatrice Cynthia Walter

doi:10.33096/woh.vi.3083

Azizul Berlyansah Universitas Internasional Batam, Batam, Riau Islands 29456, Indonesia.
Andreanyta Meliala Department of Physiology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281, Indonesia.
Paramita Narwidina Clinical Nutrition Research Group, Yogyakarta 55132, Indonesia.
Wira Mondana Department of Pediatrics, School of Medicine, Universitas Internasional Batam, Batam, Riau Islands 29456, Indonesia.
Beatrice Cynthia Walter Department of Clinical Nutrition, School of Medicine, Universitas Internasional Batam, Batam, Riau Islands 29456, Indonesia

DOI: https://doi.org/10.33096/woh.vi.3083

Keywords: Calophyllum inophyllum seed flour, fatty acid, amino acids, functional food, metabolic diseases

Abstract

Metabolic diseases such as obesity, type 2 diabetes mellitus, and dyslipidemia are health challenges, with over one billion people affected by obesity and 589 million adults living with diabetes. This study evaluated the antioxidant activity, anthocyanin content, amino acid profile, proximate composition, and fatty acid (FFA) profile of Calophyllum inophyllum seed flour as a functional food ingredient. Seeds were collected from the Batam Botanical Garden, Indonesia, and processed by freeze-drying. Antioxidant activity was determined using the DPPH assay (IC₅₀), anthocyanin content was measured spectrophotometrically, and proximate composition, amino acid profile, and FFA composition were analyzed using standardized methods, including GC-FID. All analyses were performed in triplicate (n = 3), and results were expressed as mean ± standard deviation. The flour was predominantly composed of lipids (65.26 ± 0.13%), followed by carbohydrates (21.92 ± 0.10%) and proteins (8.39 ± 0.02%), with low moisture content (2.64 ± 0.03%). The DPPH assay yielded an IC₅₀ value of 13,362.04 ± 1.70 mg/L, indicating limited free radical scavenging activity, while anthocyanins were not detected. The amino acid profile was characterized by high glutamic acid content and the presence of essential amino acids, including leucine, valine, and lysine. The fatty acid profile was dominated by unsaturated fatty acids, particularly oleic and linoleic acids. These findings indicate that Calophyllum inophyllum seed flour contains nutritionally relevant amino acids and unsaturated fatty acids, supporting its potential relevance for functional food development. Further studies are required to characterize additional bioactive compounds and evaluate their biological activities in vivo.

References

IDF. 11th Edition | 2025 Diabetes Atlas. System of Ghosts. 2025. 3–3 p.

Ruze R, Liu T, Zou X, Song J, Chen Y, Xu R, et al. Obesity and type 2 diabetes mellitus: connections in epidemiology, pathogenesis, and treatments. Front Endocrinol (Lausanne). 2023;14(April):1–23.

Linda TM, Febriarti BL, Zul D, Sofyanti N, Berliansyah A, Delfira N, et al. Isolation and characterization of endophytic bacteria from sterile leaf of Acrostichum aureum from Bengkalis Island (Riau, Indonesia) and its potency for antidiabetic. Biodiversitas. 2023;24(3):1580–8.

Linda TM, Maisyaroh DP, Berlyansah A, Tasliyah BJ, Juliantari E, Zul D, et al. Efficacy of Endophytic Bacterium Serratia marcescens B.SB 1.1 associated with Sea Fern (Acrostichum aureum L.) as an Antidiabetic Agent. J Microbiol Biotechnol. 2025;35:1–13.

Cavaliere G, Cimmino F, Trinchese G, Catapano A, Petrella L, D’Angelo M, et al. From Obesity-Induced Low-Grade Inflammation to Lipotoxicity and Mitochondrial Dysfunction: Altered Multi-Crosstalk between Adipose Tissue and Metabolically Active Organs. Antioxidants. 2023;12(6).

Bruic M, Grujic-Milanovic J, Miloradovic Z, Jovovic D, Zivkovic L, Mihailovic-Stanojevic N, et al. DNA, protein and lipid oxidative damage in tissues of spontaneously hypertensive versus normotensive rats. Int J Biochem Cell Biol [Internet]. 2021;141(April):106088. Available from: https://doi.org/10.1016/j.biocel.2021.106088

Chung HK, Kim JH, Choi A, Ahn CW, Kim YS, Nam JS. Antioxidant-Rich Dietary Intervention Improves Cardiometabolic Profiles and Arterial Stiffness in Elderly Koreans with Metabolic Syndrome. Yonsei Med J. 2022;63(1):26–33.

Aljerf L, Maad AH, Rasool S, Alajlani M. Harnessing the antioxidant and cytoprotective power of Aitchisonia rosea: phytochemical insights and mechanistic validation. BMC Plant Biol. 2025;25(1).

Hapsari S, Yohed I, Kristianita RA, Jadid N, Aparamarta HW, Gunawan S. Phenolic and flavonoid compounds extraction from Calophyllum inophyllum leaves. Arab J Chem [Internet]. 2022;15(3):103666. Available from: https://doi.org/10.1016/j.arabjc.2021.103666

Uslu K, Çavuşoğlu K, Yalçın E. Quantitative phenolic profiling and protective effects of grape seed extract on mancozeb-induced cellular and genetic toxicity. Sci Rep. 2025;15(1):1–15.

Cassien M, Mercier A, Thétiot-Laurent S, Culcasi M, Ricquebourg E, Asteian A, et al. Improving the antioxidant properties of calophyllum inophyllum seed oil from french polynesia: Development and biological applications of resinous ethanol-soluble extracts. Antioxidants. 2021;10(2):1–23.

Bababe AB, Madu FA, Abacha YZ, Iliya I, Yesufu HB, Mohammed GT, et al. Isolation, Characterization, and Evaluation of Anticancer and Antioxidant Bioactive Compounds from Calophyllum inophyllum Leaves. Int J Res. 2025;12(03):780–803.

Ojah EO, Moronkola DO, Osamudiamen PM. Corrigendum: Antioxidant assessment of characterised essential oils from Calophyllum inophyllum Linn using 2,2-diphenyl-1- picrylhydrazyl and hydrogen peroxide methods. J Med Plants Econ Dev. 2021;5(1):2–4.

Ruangsuriya J, Sichaem J, Tantraworasin A, Saeteng S, Wongmaneerung P, Inta A, et al. Phytochemical Profiles and Anticancer Effects of Calophyllum inophyllum L . Extract Relating to Reactive Oxygen Species Modulation on Patient-Derived Cells from Breast and Lung Cancers. 2023;2023.

Di Nunzio M, Picone G, Pasini F, Chiarello E, Caboni MF, Capozzi F, et al. Olive oil by-product as functional ingredient in bakery products. Influence of processing and evaluation of biological effects. Food Res Int [Internet]. 2020;131(December 2019):108940. Available from: https://doi.org/10.1016/j.foodres.2019.108940

Kaikkonen JE, Jula A, Viikari JSA, Juonala M, Hutri-kähönen N, Kähönen M, et al. Associations of Serum Fatty Acid Proportions with Obesity , Insulin Resistance , Blood Pressure , and Fatty Liver : The Cardiovascular Risk in Young Finns Study. J Nutr [Internet]. 2021;151(4):970–8. Available from: https://doi.org/10.1093/jn/nxaa409

Ateye MD, Ali AM, Hassan SM, Jama HM, Abdurahman AS. Comparative analysis of nutritional composition: proximate and mineral content of soybean varieties. Discov Food. 2025;5(1).

Kaseke T, Opara UL, Fawole OA. Fatty acid composition, bioactive phytochemicals, antioxidant properties and oxidative stability of edible fruit seed oil: effect of preharvest and processing factors. Heliyon [Internet]. 2020;6(9):e04962. Available from: https://doi.org/10.1016/j.heliyon.2020.e04962

Vicentini-polette CM, Rodolfo P, Bernardo C, Lopes A, Oliveira D. Food Chemistry : X Determination of free fatty acids in crude vegetable oil samples obtained by high-pressure processes. Food Chem X [Internet]. 2021;12:100166. Available from: https://doi.org/10.1016/j.fochx.2021.100166

Li H, Yu W, Yang T. Impact of macronutrient composition in low-glycemic biscuits on gut microbiota and metabolic profiles: Insights from in vitro fermentation. Food Biosci [Internet]. 2025;69(March):106947. Available from: https://doi.org/10.1016/j.fbio.2025.106947

Fratianni F, Coppola F, Tavaniello S, Ombra MN, De Giulio B, D’Agostino N, et al. Fatty Acid Profile and Some Useful Biological Aspects of Borage, Calophyllum, and Prickly Pear Seed Oils: Implications for Health and Dietary Use. Antioxidants. 2025;14(6):1–18.

Li G, Li Z, Liu J. Amino acids regulating skeletal muscle metabolism: mechanisms of action, physical training dosage recommendations and adverse effects. Nutr Metab. 2024;21(1):1–14.

Kolaczkowski OM, Goodson BA, Montenegro Vazquez V, Jia J, Qadir Bhat A, Kim TH, et al. Synergistic Role of Amino Acids in Enhancing mTOR Activation Through Lysosome Positioning. bioRxiv [Internet]. 2024;2024.10.12.618047. Available from: https://pmc.ncbi.nlm.nih.gov/articles/PMC11482915/

Gorissen SHM, Crombag JJR, Senden JMG, Waterval WAH, Bierau J, Verdijk LB, et al. Protein content and amino acid composition of commercially available plant-based protein isolates. Amino Acids [Internet]. 2018;50(12):1685–95. Available from: https://doi.org/10.1007/s00726-018-2640-5

Wa Y, Zhao X, Zhang C, Qu H, Chen D, Chen X, et al. The transcriptional regulation effects of histidine, isoleucine and glutamate on free exopolysaccharide biosynthesis of Streptococcus thermophilus 937. Front Microbiol. 2024;15(January):1–15.

Li CWD, Herpich C, Haß U, Kochlik B, Weber D, Grune T, et al. Essential amino acids and branched-chain amino acids are associated with skeletal muscle and inflammatory parameters in older age. Biogerontology [Internet]. 2025;26(2):1–10. Available from: https://doi.org/10.1007/s10522-025-10206-1

Domić J, van Loon LJC, Siebelink E, Borgonjen-Van den Berg KJ, de Groot LCPGM, Grootswagers P. The amino acid composition of commercially available vegan meat and dairy analogues. Br J Nutr. 2025;134(5):377–89.

Anjukam E, Ramesh M, Rajalakshmi A, Kavitha K, Prakash M, Suresh G, et al. Molecular characterization of potent antibacterial compound 4-(3-methylazetidin-1-yl)pentan-2-ol from Calophyllum inophyllum seed oil. Biocatal Agric Biotechnol [Internet]. 2023;47(November 2022):102617. Available from: https://doi.org/10.1016/j.bcab.2023.102617

Nalimanana NR, Tombozara N, Razafindrakoto ZR, Andrianjara C, Ramanitrahasimbola D. Anti-inflammatory and analgesic properties, and toxicity of the seed’s ethanol extract of Calophyllum inophyllum L. from the eastern region of Madagascar. South African J Bot. 2022;150:466–72.

Pardeshi SR, Deshmukh NS, Telange DR, Nangare SN, Sonar YY, Lakade SH, et al. Process development and quality attributes for the freeze-drying process in pharmaceuticals, biopharmaceuticals and nanomedicine delivery: a state-of-the-art review. Futur J Pharm Sci [Internet]. 2023;9(1). Available from: https://doi.org/10.1186/s43094-023-00551-8

Ojah EO, Moronkola DO, Osamudiamen PM. Corrigendum: Antioxidant assessment of characterised essential oils from Calophyllum inophyllum Linn using 2,2-diphenyl-1- picrylhydrazyl and hydrogen peroxide methods. J Med Plants Econ Dev. 2021;5(1).

Li S, Liu F, Zhang S, Sun X, Li X, Yue Q, et al. Lavender Exosome-Like nanoparticles attenuate UVB- Induced Photoaging via miR166- Mediated inflammation and collagen regulation. 2025;1–15.

Bulanov NМ, Suvorov AY, Blyuss OB, Munblit DB, Butnaru D V, Nadinskaia MY, et al. Basic principles of descriptive statistics in medical research Основные принципы применения описательной статистики в медицинских исследованиях. 2021;12(3):4–16.

Du M, Xie J, Gong B, Xu X, Tang W, Li X, et al. Extraction, physicochemical characteristics and functional properties of Mung bean protein. Food Hydrocoll [Internet]. 2018;76:131–40. Available from: http://dx.doi.org/10.1016/j.foodhyd.2017.01.003

Xie Y, Wang Y, Jin X, Zhang X, Yang R. Pumpkin Seed Proteins: The Potentially Alternative Protein Supplements for Food Applications. Foods. 2025;14(22):1–33.

van den Berg LA, Jurriaan JM, Mensink M, Wanders AJ. Protein quality of soy and the effect of processing: A quantitative review. Front Nutr. 2022;1–14.

Alejandro Ruiz FE, Ortega Jácome JF, Mora JR, Landázuri AC, Vásconez Duchicela P, Vásconez Espinoza J, et al. Comprehensive characterization and valorization potential of Amazonian Sacha inchi (Plukenetia volubilis L.) seeds, oil, and oilcake by-products for sustainable food applications. Front Nutr. 2025;12.

Usman I, Saif H, Imran A, Afzaal M, Saeed F, Azam I, et al. Innovative applications and therapeutic potential of oilseeds and their by-products: An eco-friendly and sustainable approach. Food Sci Nutr. 2023;11(6):2599–609.

Ahmed I, Chatha SAS, Iftikhar N, Farooq MF, Zulfiqar H, Ali S, et al. Nutritional quality of selected commercially available seed oils and effect of storage conditions on their oxidative stability. PLoS One [Internet]. 2024;19(10 October):1–18. Available from: http://dx.doi.org/10.1371/journal.pone.0308117

Akbar U, Mondol MSA, Singh J, Rasane P, Nanda V, Abdi G, et al. Effects of various packaging materials and temperature conditions on the storage stability of Zea mays L. (baby corn) powder. Appl Food Res [Internet]. 2025;5(1):100913. Available from: https://doi.org/10.1016/j.afres.2025.100913

Muflihah YM, Gollavelli G, Ling YC. Correlation study of antioxidant activity with phenolic and flavonoid compounds in 12 indonesian indigenous herbs. Antioxidants. 2021;10(10):1–15.

Xiang Y, Xiang M, Mao Y, Huang L, He Q, Dong Y. Insights into structure-antioxidant activity relationships of polyphenol-phospholipid complexes: The effect of hydrogen bonds formed by phenolic hydroxyl groups. Food Chem. 2025;485(April).

Vittaya L, Chalad C, Janyong S, Leesakul N. Bioactive Potential of Calophyllum inophyllum: Phytochemical Profiles, Biological Activities, and In Silico Pharmacokinetic Predictions. Phyton-International J Exp Bot. 2025;94(12):4091–115.

La JW, Kim MJ, Lee JH. Evaluation of solvent effects on the DPPH reactivity for determining the antioxidant activity in oil matrix. Food Sci Biotechnol [Internet]. 2021;30(3):367–75. Available from: https://doi.org/10.1007/s10068-020-00874-9

Liu J, Zhao L, Cai H, Zhao Z, Wu Y, Wen Z, et al. Antioxidant and Anti-Inflammatory Properties of Rubber Seed Oil in Lipopolysaccharide-Induced RAW 267.4 Macrophages. Nutrients. 2022;14(7).

Vincek V, Vincek D, Makovec M, Šamec D. Intra-annual and regional variability of the oleic acid and linoleic acid content of pumpkin seed oil over a 10-year period. J Food Compos Anal. 2024;135(April).

Liu H, Li J, Xu C, Liu H, Zhao Z. Unravelling the molecular regulation network of carbon metabolism and lipid metabolism during seed development in Akebia trifoliata via integrated multi-omics analysis. Sci Rep. 2024;14(1):1–16.

Pothinam S, Putpim C, Siriwoharn T, Jirarattanarangsri W. Effects of Perilla Seed Oil on Blood Lipids, Oxidative Stress, and Inflammation in Hyperlipidemic Rats. Foods. 2025;14(8).

Tamura K, Kitazawa H, Sugita S, Hashizume K, Iwashita M, Ishigami T, et al. Tyrosine Is a Booster of Leucine-Induced Muscle Anabolic Response. Nutrients. 2024;16(1):1–15.

Zhao YX, Tong L, Zhang GM, Zhao XH, Sa YP, Liu Y, et al. L-Arginine Supplementation Improves Vascular Endothelial Dysfunction Induced by High-Fat Diet in Rats Exposed to Hypoxia. Wilderness Environ Med. 2020;31(4):400–6.

Teixeira WF, Fagan EB, Soares LH, Umburanas RC, Reichardt K, Neto DD. Foliar and seed application of amino acids affects the antioxidant metabolism of the soybean crop. Front Plant Sci. 2017;8(March):1–14.

Radika MK, Viswanathan P, Anuradha C V. Nitric oxide mediates the insulin sensitizing effects of β-sitosterol in high fat diet-fed rats. Nitric Oxide - Biol Chem [Internet]. 2013;32:43–53. Available from: http://dx.doi.org/10.1016/j.niox.2013.04.007

Vera Zambrano M, Dutta B, Mercer DG, MacLean HL, Touchie MF. Assessment of moisture content measurement methods of dried food products in small-scale operations in developing countries: A review. Trends Food Sci Technol [Internet]. 2019;88(July 2018):484–96. Available from: https://doi.org/10.1016/j.tifs.2019.04.006

Su J, Geng Y, Yao J, Huang Y, Ji J, Chen F, et al. Quinone-mediated non-enzymatic browning in model systems during long-term storage. Food Chem X [Internet]. 2022;16(October):100512. Available from: https://doi.org/10.1016/j.fochx.2022.100512

Mattioli S, Dal Bosco A, Castellini C, Falcinelli B, Sileoni V, Marconi O, et al. Effect of heat- and freeze-drying treatments on phytochemical content and fatty acid profile of alfalfa and flax sprouts. J Sci Food Agric. 2019;99(8):4029–35.

Seke F, Manhivi VE, Shoko T, Slabbert RM, Sultanbawa Y, Sivakumar D. Effect of freeze drying and simulated gastrointestinal digestion on phenolic metabolites and antioxidant property of the natal plum (Carissa macrocarpa). Foods. 2021;10(6).