Subhan Rullyansyah, Idha Kusumawati, Intan Safinar Ismail, Djoko Agus Purwanto, Dewi Isadiartuti, Muhammad Safwan Bin Ahamad Bustaman, Muhammad Afiq Bin Ngadni, Rizky Rafi Rahmawan, Ananda Permata Fitri, Charlyna Veronika Puspitasari Pattymahu
Objectives: Curcuma longa L. (CL) exhibits potent anti-inflammatory and analgesic properties suitable for pharmacopuncture; however, the clinical application of CL is severely hindered by poor aqueous solubility. Moreover, traditional ethanol-based extracts are unsuitable for safe parenteral administration due to the risk of tissue necrosis and neurotoxicity at sensitive acupoint sites. Thus, no systematic safety evaluation of water-soluble CL extract (CLE) formulations for injection has yet been conducted. Methods: Spray-dried CLE (SDCLE) was prepared by spray-drying CLE with lactose (1:9 w/w) as the microencapsulation agent. The curcuminoid composition was quantified using a validated high-performance thin-layer chromatography (HP-TLC) method. The solubility of SDCLE was compared with that of native CLE (NCLE). Acute toxicity (LC50), survival, and locomotor behavior were determined using probit regression and automated EthoVision XT tracking following a 96-hour exposure of adult zebrafish to SDCLE (0-500 mg/L). Results: SDCLE, formulated to contain 10% (w/w) NCLE, exhibited a curcumin content of 0.148 ± 0.02 mg/g. This formulation showed markedly enhanced aqueous solubility (91.30% at a 1:30 dilution) compared with NCLE alone (55.78% at the same dilution), yielding clear, homogeneous solutions suitable for parenteral administration. Post-exposure acute toxicity assessment in zebrafish revealed a time-dependent decrease in LC50 values, from 469.96 mg/L at 24 h to 288.13 mg/L at 96 h. Acute toxicity analysis identified a no-observed-effect concentration (NOEC) of 144.07 mg/L and a lowest-observed-effect concentration (LOEC) of 187.28 mg/L. Exposure to concentrations ≤ 200 mg/L resulted in 95% survival and preserved normal locomotor activity, whereas concentrations ≥ 300 mg/L induced pronounced neurobehavioral suppression, characterized by a 35% reduction in swimming distance and a six-fold increase in inactivity. Conclusion: Spray-drying overcame the solubility limitations of CLE, enabling aqueous reconstitution for pharmacopuncture applications. A 96-hour LC50 of 288.13 mg/L was employed to define mortality, while neurobehavioral endpoints indicated toxicity at sublethal concentrations, supporting the safety of concentrations ≤ 200 mg/L for rational SDCLE dose selection. Copyright © Korean Pharmacopuncture Institute This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Doctoral Program in Pharmaceutical Sciences, Faculty of Pharmacy, Universitas Airlangga, Surabaya, Indonesia; Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Negeri Surabaya, Surabaya, Indonesia; Department of Pharmaceutical Science, Faculty of Pharmacy, Universitas Airlangga, Surabaya, Indonesia; Ethnomedicine and Indonesian Traditional Medicine Development Research Center (E-ITMeD RC), Faculty of Pharmacy, Universitas Airlangga, Surabaya, Indonesia; HPTLC Association Indonesia Chapter, Jakarta, Indonesia; Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, UPM Serdang, Selangor, Malaysia; Natural Medicines and Product Research Laboratory (NaturMeds), Institute of Bioscience (IBS), Universiti Putra Malaysia, UPM Serdang, Selangor, Malaysia; Magister Program, Faculty of Pharmacy, Universitas Airlanga, Surabaya, Indonesia
