DOI: https://doi.org/10.14710/jksa.29.3.217-226

Optimizing Maceration Extraction of Myristicin and Methyl Eugenol from Nutmeg (Myristica fragrans Houtt.): An RSM Box–Behnken Approach

1Chemistry Science Doctoral Program, Department of Chemistry, Faculty of Mathematics and Natural Sciences, IPB University, Bogor, Indonesia

2Chemistry Education Study Program, Faculty of Islamic Education and Teacher Training, Syarif Hidayatullah State Islamic University Jakarta, Indonesia

3Department of Chemistry, Faculty of Mathematics and Natural Science, IPB University, Bogor, Indonesia

4 Tropical Biopharmaca Research Center, International Research Institute of Food, Nutrition, and Health, IPB University, Bogor, Indonesia

5 Study Program of Statistics and Data Science, IPB University, Bogor, Indonesia

6 Research Center for Metallurgy, National Research and Innovation Agency (BRIN), South Tangerang, Indonesia

View all affiliations
Received: 25 Nov 2025; Revised: 3 Mar 2026; Accepted: 30 Mar 2026; Published: 22 Apr 2026.
Open Access Copyright 2026 Jurnal Kimia Sains dan Aplikasi under http://creativecommons.org/licenses/by-sa/4.0.

Citation Format:
Cover Image
Abstract
Myristicin is the key marker of nutmeg (Myristica fragrans) quality, while methyl eugenol is an undesirable compound requiring strict monitoring. This study aimed to optimize the extraction conditions of these compounds from nutmeg seeds and mace as a pretreatment step prior to electrochemical quantification. Optimization was performed using Response Surface Methodology (RSM) with a Box–Behnken Design (BBD), considering extraction time (h), ethanol concentration (%), and the sample-to-solvent ratio (g/mL) as independent variables. The analytes were quantified by Gas Chromatography–Mass Spectrometry (GC-MS), and the experimental data were modeled using Design Expert software. Regression models indicated that ethanol concentration and solvent volume significantly influenced extraction yields, while extraction time had a statistically minimal effect (p-values > 0.05, small effect sizes). Nevertheless, practical differences were observed between matrices: nutmeg seeds reached optimal extraction within 2 hours, whereas mace required 17 hours, likely due to its higher essential oil content and fibrous structure, which slows diffusion kinetics. Thus, although time effects were not statistically significant, extended extraction was practically preferred for mace to ensure adequate recovery. Response definitions were specified as follows: Y1 = myristicin concentration (mg/mL), Y2 = methyl eugenol concentration (mg/mL), and % extraction yield. The optimal extraction conditions were achieved with 90% ethanol and 50 mL solvent, maximizing analyte detectability while maintaining reproducibility across seed and mace extracts. These optimized ethanolic matrices are compatible with subsequent electrochemical quantification, given their aqueous/ethanolic composition and anticipated sensitivity of electrochemical detection.
Fulltext View|Download Email colleagues
Keywords: myristicin; methyl eugenol; Response Surface Methodology; Gas Chromatography-Mass Spectrometry; Design Expert
Funding: Directorate General of Research and Development of the Ministry of Higher Education, Science, and Technology in accordance with the Research Program Implementation Contract for Fiscal Year 2025 under contract 006/C3/DT.05.00/PL/2025

Article Metrics:

Article Info
Section: Research Articles
Language : EN
Most cited articles
Lempung Terpilar Silika untuk Penghilangan Fenol dalam Air Innovation of Carbon from Pectin Templated in Fabrication of Interlayer-free Silica-Pectin Membrane Sintesis Metilisoeugenol dari Metileugenol Menggunakan Katalis KOH dalam Etanol Asam Anakardat dari Kulit Biji Jambu Mete (Anacardium Occidentale L) yang Mempunyai Aktivitas Sitotoksik Dimerisasi Metilisoeugenol dengan Katalis HCl More cited articles
  1. Chia-Jung Lee, Chun-Wei Huang, Lih-Geeng Chen, Ching-Chiung Wang, (+)-Erythro-∆8'-7S,8R-Dihydroxy-3,30,50-Trimethoxy-8-O-40-Neolignan, an Anti-Acne Component in Degreasing Myristica fragrans Houtt, Molecules, 25, 19, (2020), 4563 https://doi.org/10.3390/molecules25194563
  2. Monika Vasantrao Padol, Prashanth Vishwakarma, Arun Suresh Dodamani, Anup Wamanrao Gore, Karina Shyamlal Chachlani, Shradha Pramod Kharkar, Comparative evaluation of nutmeg mouthwash and 0.2% chlorhexidine gluconate mouthwash on halitosis and plaque control: A randomized clinical trial, Journal of Indian Society of Periodontology, 26, 4, (2020), 384-389 https://doi.org/10.4103/jisp.jisp_868_20
  3. Hailan Bao, Qi Muge, Anticancer effect of myristicin on hepatic carcinoma and related molecular mechanism, Pharmaceutical Biology, 59, 1, (2021), 1126-1132 https://doi.org/10.1080/13880209.2021.1961825
  4. Elisa Frederico Seneme, Daiane Carla Dos Santos, Carolina Afonso de Lima, Ícaro Augusto Maccari Zelioli, Juliana Mozer Sciani, Giovanna Barbarini Longato, Effects of Myristicin in Association with Chemotherapies on the Reversal of the Multidrug Resistance (MDR) Mechanism in Cancer, Pharmaceuticals, 15, 10, (2022), 1233 https://doi.org/10.3390/ph15101233
  5. Sawsan S. Al-Rawi, Ahmad Hamdy Ibrahim, Heshu Jalal Ahmed, Zhikal Omar Khudhur, Therapeutic, and pharmacological prospects of nutmeg seed: A comprehensive review for novel drug potential insights, Saudi Pharmaceutical Journal, 32, 6, (2024), 102067 https://doi.org/10.1016/j.jsps.2024.102067
  6. Muhammad Imran, Abdul Haleem Shah, Niamat Ullah, Suliman Yousef Alomar, Abdur Rehman, Naeem Ur Rehman, Asif Nawaz, Rabia Baloch, Ali Zaman, Hafiz Abdul Rafey, Khizar Abbas, Adnan Amin, Integrated computational analysis, in vitro, in vivo investigation on Myristica fragrans Houtt. essential oils for potential anti rheumatic activities, Journal of King Saud University - Science, 36, 5, (2024), 103177 https://doi.org/10.1016/j.jksus.2024.103177
  7. Fuat Karakuş, Zübeyde Tanrıverdi, Burak Kuzu, In Silico Network Toxicology, Molecular Docking, and Multi-Level Bioinformatics Reveal Methyl Eugenol-Induced Hepatocellular Carcinoma Mechanisms in Humans, Cancer Medicine, 14, 10, (2025), 1-11 https://doi.org/10.1002/cam4.70768
  8. Xiaoling Zhang, Susan P. Felter, Anne Marie Api, Kaushal Joshi, Dan Selechnik, A Cautionary tale for using read-across for cancer hazard classification: Case study of isoeugenol and methyl eugenol, Regulatory Toxicology and Pharmacology, 136, (2022), 105280 https://doi.org/10.1016/j.yrtph.2022.105280
  9. Rudi Heryanto, Siti Sa’diah, Waras Nurcholis, Application of two-level factorial design in optimization of maceration extraction of Curcuma longa curcuminoids extracts, Journal of Applied Biology & Biotechnology, 12, 5, (2024), 143-148 https://doi.org/10.7324/JABB.2024.179132
  10. Anthonin Gori, Benjamin Boucherle, Aurélien Rey, Maxime Rome, Nicola Fuzzati, Marine Peuchmaur, Development of an innovative maceration technique to optimize extraction and phase partition of natural products, Fitoterapia, 148, (2021), 104798 https://doi.org/10.1016/j.fitote.2020.104798
  11. Manoj Madanahalli Ramesh, Nischith Sirawase Shankar, Annegowda Hardur Venkatappa, Driving/Critical Factors Considered During Extraction to Obtain Bioactive Enriched Extracts, Pharmacognosy Reviews, 18, 35, (2024), 68-81 https://doi.org/10.5530/phrev.2024.18.7
  12. Alexander Weremfo, Samuel Abassah-Oppong, Felix Adulley, Kwabena Dabie, Salifu Seidu-Larry, Response surface methodology as a tool to optimize the extraction of bioactive compounds from plant sources, Journal of the Science of Food and Agriculture, 103, 1, (2023), 26-36 https://doi.org/10.1002/jsfa.12121
  13. Ivana Karabegoić, Sandra Stamenković Stojanović, B. Danilović, Optimization of food-grade bioactive compound extraction from plant sources by response surface methodology approach: A comprehensive review, Maso International – Journal of Food Science and Technology, 13, 1, (2023), 31-47 https://doi.org/10.2478/mjfst-2023-0004
  14. Syeda Nashvia Adin, Isha Gupta, Mohd Aqil, Mohd Mujeeb, Abdul Ahad, BBD Driven Optimization of Extraction of Therapeutically Active Xanthanoid Mangiferin from Mangifera indica L. Leaves and its Antioxidant Activity, Pharmacognosy Research, 15, 1, (2022), 84-93 https://doi.org/10.5530/097484900279
  15. Vicente Amirpasha Tirado-Kulieva, Manuel Sánchez-Chero, Mario Villegas Yarlequé, Gretell Fiorela Villegas Aguilar, Gilberto Carrión-Barco, Abraham Guillermo Ygnacio Santa Cruz, José Sánchez-Chero, An Overview on the Use of Response Surface Methodology to Model and Optimize Extraction Processes in the Food Industry, Current Research in Nutrition and Food Science, 9, 3, (2021), 745-754 https://doi.org/10.12944/CRNFSJ.9.3.03
  16. Filipa Paulo, Loleny Tavares, Lúcia Santos, Response Surface Modeling and Optimization of the Extraction of Phenolic Antioxidants from Olive Mill Pomace, Molecules, 27, 23, (2022), 8620 https://doi.org/10.3390/molecules27238620
  17. Dara Juliana, Bambang Pontjo Priosoeryanto, Waras Nurcholis, Optimization of Cardamom Fruit (Amomum compactum) Extraction Focused on Total Terpenoid Extraction and Cytotoxic Activity Using Response Surface Methodology, Jurnal Jamu Indonesia, 8, 2, (2023), 57-69 https://doi.org/10.29244/jji.v8i2.328
  18. Irmanida Batubara, Yanico Hadi Prayogo, Stéphane Dumarcay, Christine Gerardin, Philippe Gerardin, Wayan Darmawan, Ika Resmeiliana, Maria Celeste Ruiz, Auliya Ilmiawati, Harlinda Kuspradini, Mohamad Rafi, Phytochemical Profile and In Vitro – In Silico Antibacterial Activity of Melia azedarach Leaf and Twig Extracts Obtained Using Solvents of Different Polarities, Sci, 7, 4, (2025), 167 https://doi.org/10.3390/sci7040167
  19. Irmanida Batubara, Indah Fajarwati, Y. W. Sari, Gilles J. Guillemin, I. Maulidya, Dinda Iryawati, W. Wahyuni, Evaluation of acute and sub-acute toxicity of Curcuma aeruginosa Roxb. essential oil in Sprague Dawley rats, Research in Pharmaceutical Sciences, 20, 3, (2025), 456-468 https://doi.org/10.4103/RPS.RPS_9_24
  20. Saengnapa Champasuri, Arunporn Itharat, Bioactivities of Ethanolic Extracts of Three Parts (Wood, Nutmeg and Mace) from Myristica fragrans Houtt., Journal of the Medical Association of Thailand, 99, (2016), S124-S130
  21. Halid Makić, Samira Hotić, Elvisa Hodžić, Nenad Stojanović, Jasmina Ibrahimpašić, Samira Dedić, Emina Ćehajić Gradinović, Influence of Maceration, Solvent Types, and Extraction Durations on the Yield of Milk Thistle Seeds (Silybum Marianum) Extraction, Technologica Acta, 17, 1, (2024), 67-76 https://doi.org/10.51558/2232-7568.2023.17.1.67
  22. Manh Tuan Ha, Ngoc Khanh Vu, Thu Huong Tran, Jeong Ah Kim, Mi Hee Woo, Byung Sun Min, Phytochemical and pharmacological properties of Myristica fragrans Houtt.: an updated review, Archives of Pharmacal Research, 43, 11, (2020), 1067-1092 https://doi.org/10.1007/s12272-020-01285-4
  23. Mega Ferdina Warsito, A Review on Chemical Composition, Bioactivity, and Toxicity of Myristica fragrans Houtt. Essential Oil, Indonesian Journal of Pharmacy, 32, 3, (2021), 304-313 https://doi.org/10.22146/ijp.1271
  24. Khairan Khairan, Meutia Faradilla, Binawati Ginting, Elly Sufriadi, Adinda Gusti Vonna, Muslim Akmal, Amalia, Ernawati, Indra, Hizir Sofyan, Muhammad Diah, Syaifullah Muhammad, Isolation of Nutmeg Essential Oil (Myristica fragrans houtt) from Aceh Indonesia and Their Antioxidant and Antibacterial Activities, Indonesian Journal of Pharmacy, 35, 3, (2024), 425-436 https://doi.org/10.22146/ijp.8399
  25. Ismiyarto, Ngadiwiyana, Rani Mustika, Isolasi, Identifikasi Minyak Atsiri Fuli Pala (Myristica fragrans) dan Uji Aktivitas Sebagai Larvasida, Jurnal Kimia Sains dan Aplikasi, 12, 1, (2009), 23-30 https://doi.org/10.14710/jksa.12.1.23-30
  26. Nikolaos Nenadis, Maria Papapostolou, Maria Z. Tsimidou, Suggestions on the Contribution of Methyl Eugenol and Eugenol to Bay Laurel (Laurus nobilis L.) Essential Oil Preservative Activity through Radical Scavenging, Molecules, 26, 8, (2021), 2342 https://doi.org/10.3390/molecules26082342
  27. Lidwina Angelica Soetantijo, Hendrik O. Lintang, Heriyanto, Mitha Ayu Pratama Handojo, Tatas Hardo Panintingjati Brotosudarmo, Optimization of Adsorption and Desorption Time in the Extraction of Volatile Compounds in Brewed Java Arabica Coffee Using the HS-SPME/GC-MS Technique, Jurnal Kimia Sains dan Aplikasi, 25, 2, (2022), 49-55 https://doi.org/10.14710/jksa.25.2.49-55
  28. Erik V. Petersson, Jiayin Liu, Per J. R. Sjöberg, Rolf Danielsson, Charlotta Turner, Pressurized Hot Water Extraction of anthocyanins from red onion: A study on extraction and degradation rates, Analytica Chimica Acta, 663, 1, (2010), 27-32 https://doi.org/10.1016/j.aca.2010.01.023
  29. Nashwa F. S. Morsy, A comparative study of nutmeg (Myristica fragrans Houtt.) oleoresins obtained by conventional and green extraction techniques, Journal of Food Science and Technology, 53, 10, (2016), 3770-3777 https://doi.org/10.1007/s13197-016-2363-0
  30. Muliadi, Mudzuna Quraisyah Basimin, Ahmad Muchsin Jayali, Density Functional Theory for QSAR Antioxidant Compound Myristicin Derivatives, Indonesian Journal of Chemical Research, 9, 1, (2021), 49-56 https://doi.org/10.30598//ijcr.2021.9-mul
  31. Salony Azam Sheikh, Vallamkondu Manasa, Palak Daga, Ajay W. Tumaney, Does fixed oil impart more antioxidant potential to spices than essential oils? A comparative study with seed (nutmeg) and aril (mace) oil from the spice myristica fragrans, Journal of Food Science and Technology, 62, 11, (2025), 2173-2182 https://doi.org/10.1007/s13197-024-06164-0
  32. Nicky Rahmana Putra, Ahmad Hazim Abdul Aziz, Hasmadi Mamat, Dwila Nur Rizkiyah, Mohd Azizi Che Yunus, Irianto Irianto, Lailatul Qomariyah, Green extraction of nutmeg (Myristica fragrans) phytochemicals: Prospective strategies and roadblocks, Open Agriculture, 9, 1, (2024), 20220285 https://doi.org/10.1515/opag-2022-0285

Last update:

No citation recorded.

Last update: 2026-04-22 12:04:43

No citation recorded.