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Sequential extraction protocol for bio-calcium from saltwater crocodile (Crocodylus porosus) bone: Physicochemical and molecular studies
(2026-06-01)
Yarnpakdee, Suthasinee
;
Senphan, Theeraphol
;
Benjakul, Soottawat
;
Sriket, Chodsana
Saltwater crocodile ( Crocodylus porosus ; SC) farming generates 45% bone waste, creating environmental challenges but also representing an underutilized source of calcium. This study presents the first comprehensive protocol for extracting bio-calcium from SC bone through sequential processing: autoclaving, alkaline soaking, ethanol extraction, and hydrogen peroxide bleaching. The extraction process yielded 16.94% bio-calcium with a high mineral density (26.37% Ca and 13.83% P) and an average particle size of 6.77 μm. XRD confirmed the preserved hydroxyapatite structure, while FTIR revealed the complete elimination of the organic matrix in the resulting bio-calcium. SEM-EDS demonstrated uniform elemental distribution with no detectable heavy metals. Remarkably, in vitro calcium bioavailability reached 15.24%, representing a 78% enhancement over fish bone bio-calcium and doubling synthetic supplement performance. Preserved collagen-derived amino acids facilitate calcium-peptide chelation, explaining the superior absorption. The developed valorization process converts aquaculture waste into high-value ingredients with potential applications in food and pharmaceuticals.
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Improving calcium bioavailability from fish bone waste: the role of sodium hydroxide in bio‑calcium extraction from tilapia (Oreochromis niloticus) bones
(2025-10-01)
Senphan, Theeraphol
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Mungmueang, Natthapong
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Choommongkol, Vachira
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Sriket, Chodsana
;
Kishimura, Hideki
Calcium deficiency is a global health issue motivating the search for sustainable calcium sources from aquaculture waste. This study investigated sodium hydroxide (NaOH) concentration effects on bio‑calcium quality and bioavailability extracted from tilapia (Oreochromis niloticus) bones. Tilapia bones were treated with 0.5 M, 1 M, and 2 M NaOH solutions for 30 min compared to untreated controls. Increasing NaOH concentrations decreased yield while improving brightness, ash content, and calcium bioavailability. The 2 M NaOH treatment produced bio‑calcium with highest calcium bioavailability (8.57 %), surpassing both control (7.26 %) and commercial calcium carbonate (0.72 %) by 12-fold. Higher NaOH concentrations reduced moisture, protein, and fat contents while increasing hydroxyproline and decreasing lipid oxidation. SEM showed smoother surfaces with homogeneous pores in 2 M NaOH-treated samples. ATR-FTIR and EDS confirmed consistent hydroxyapatite structure. This optimization provides a sustainable approach for converting fish waste into high-bioavailability calcium supplements, supporting circular economy in aquaculture while addressing global calcium deficiency.
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Synthesis of soluble calcium compound from skipjack tuna bones using edible weak acids
(2022-06-01)
Aenglong, Chakkapat
;
Wang, Yu Ming
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Limpawattana, Maruj
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Sukketsiri, Wanida
;
Tang, Qing Juan
This study aimed to enhance the water solubility of calcium from tuna (Katsuwonus pelamis) bones (TB). Calcium hydroxyapatite (HA) was obtained by calcination of TB. For mineral profiles of TB and HA, Ca had the highest content followed by P with Ca/P of 3.47 and 1.94. Mg, K, and Na as macro minerals and Cr, Cu, Mn, and Zn as micro minerals were also found. Subsequently, HCl was applied to form structured HA-chloride compound (HA-Cl) then reacted with alkaline to obtain structured HA-hydroxide compound (HA-OH). In order to enhance the water solubility, citric and lactic acid were individually reacted with HA-Cl and HA-OH to obtain structured HA-citrate compounds (HA-Cl-Ci and HA-OH-Ci) and structured HA-lactate compounds (HA-Cl-Lac and HA-OH-Lac). HA-Cl-Ci had the highest water solubility (93.34 ± 0.45%) followed by HA-OH-Lac (90.35 ± 0.23%) with Ca/P ratio of 2.28 and 2.24, respectively. HA-Cl-Ci and HA-OH-Lac demonstrated the similar mineral profiles to those of HA but with different contents. The results of FT-IR indicated the carboxyl group of acid played an important role in binding Ca<sup>2+</sup>. X-ray diffractograms indicated that calcined bone was HA whereas the major composition of HA-Cl-Ci and HA-OH-Lac was related to calcium citrate tetrahydrate and calcium lactate pentahydrate, respectively.
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Bioactive alginate/carrageenan/calcium silicate porous scaffolds for bone tissue engineering
(2021-03-01)
Sathain, Ammara
;
Monvisade, Pathavuth
;
Siriphannon, Punnama
Porous bioactive alginate/carrageenan/calcium silicate scaffolds for bone tissue engineering were fabricated. The scaffolds were prepared by dispersing the synthesized calcium silicate in an aqueous solution of alginate and carrageenan at 90 °C. The scaffolds were shaped by freeze-drying and further crosslinked by 0.5, 1.0 and 1.5 M CaCl<inf>2</inf> for 60 and 120 min. The scaffolds crosslinked by 1.5 M CaCl<inf>2</inf> for 120 min achieved the highest in vitro dimension stability. The formation of hydroxyapatite crystals was observed on the scaffolds surface after soaking in simulated body fluid (SBF) at 37 °C for 7–28 days, indicating in vitro bioactivity of the scaffolds. The presence of calcium silicate could enhance not only the bioactivity, but also the mechanical properties of the scaffolds comparable to the cancellous bone. Moreover, the dimension and mechanical properties of the wet scaffolds could recover to the original after four cycles of mechanical testing at 50 % strain. The scaffolds were nontoxic to human living cells, in which the in vitro drug release behavior of the scaffold using diclofenac as a model drug was suitable for the treatment of acute inflammation after surgery. Therefore, these scaffolds were considered to be the candidate materials for bone replacement.

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