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    Development of instant germinated black rice (Maejo rice 1A) milk beverage powder fortified with Cordyceps militaris protein isolate
    (2026-08-01)
    Senphan, Theeraphol
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    Mungmueang, Natthapong
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    Jaturapanyaskul, Siri
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    Wangcharoen, Wiwat
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    Daengprok, Wichittra
    Development of instant germinated black rice (Maejo rice 1A) milk beverage powder fortified with Cordyceps militaris protein isolate was investigated. The base formula was optimized using mixture design, with Formula 8 (24% rice powder, 12% monk fruit sugar, 44% milk powder, 19.7% inulin and 0.3% Palsgaard® RecMilk) selected based on sensory evaluation. The beverage was then fortified with protein isolate at 0%, 3%, 5%, and 7% (w/w). Physicochemical properties, antioxidant activities, and sensory characteristics were evaluated. Protein fortification influenced color, water activity, viscosity, and nutritional composition, while enhancing phenolic-related compounds and antioxidant potential, indicating improved functional properties of the beverage powder. The 7% protein-fortified sample exhibited the highest antioxidant activity across all assays while maintaining acceptable thermal stability. Sensory evaluation showed that 5% protein fortification achieved optimal consumer acceptance (overall score 7.65), outperforming commercial products. The study demonstrates successful development of a functional beverage powder with enhanced nutritional and antioxidant properties through protein fortification.
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    Enhanced calcium solubility of bio-calcium powder from blackchin tilapia (Sarotherodon melanotheron) bone: Influence of alkaline pretreatment conditions
    (2026-07-01)
    Senphan, Theeraphol
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    Mungmueang, Natthapong
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    Sazedul Hoque, Md
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    Sriket, Chodsana
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    Benjakul, Soottawat
    Bio-calcium powder was prepared from blackchin tilapia ( Sarotherodon melanotheron ) bones, an invasive species by-product from surimi production in Thailand, to evaluate the effects of alkaline pretreatment on bio-calcium properties. Bones were pretreated with NaOH or KOH (1 and 2 M) at a bone-to-solution ratio of 1:10 (w/v) for 2 h, followed by ethanol washing, hydrogen peroxide bleaching, drying, and grinding to obtain bio-calcium powder. The yield on a wet weight basis was 26–28%, with water activity values of 0.33–0.34 measured at room temperature. Among all treatments, 1 M KOH gave the highest yield (28%), calcium content (26.13%), and lightness (L∗ = 88.5). Collagenous proteins and hydroxyapatite were identified by SDS-PAGE and ATR-FTIR, respectively. All bio-calcium samples showed four to five times higher in vitro calcium solubility after simulated gastrointestinal digestion (13.4-17.50%) than calcium carbonate (3.4%). The 2 M treatments exhibited the highest calcium solubility (17.2-17.50%). The 1 M KOH can be used for food fortification due to its high yield and lightness, whereas the 2 M treatments can serve as calcium supplements where higher calcium solubility is required. Blackchin tilapia bone is a potential source for bio-calcium production, with 1 M KOH suitable for food fortification and 2 M treatments more appropriate for applications requiring higher calcium solubility.
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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
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    Senphan, Theeraphol
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    Benjakul, Soottawat
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    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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    Enhanced Stability and Bioavailability of Defatted Cricket Protein Hydrolysates Encapsulated in Alginate-Coated Liposomes
    (2026-04-01)
    Chotphruethipong, Lalita
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    Benjakul, Soottawat
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    Aluko, Rotimi E.
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    Senphan, Theeraphol
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    Hutamekalin, Pilaiwanwadee
    The practical application of protein hydrolysates as functional food ingredients is frequently obstructed by their inherent structural instability. To circumvent this limitation, liposomal encapsulation has emerged as a sophisticated strategy to bolster the bioactivity and integrity of cricket-derived proteins. In this study, varying concentrations (1–4% w/v) of defatted cricket protein hydrolysate (DCPH) were integrated into vesicles composed of soy lecithin and cholesterol. The highest encapsulation efficiency (EE) was observed at a 2% DCPH loading capacity, yielding a significant result of 88.18% (p < 0.05). Subsequent coating with sodium alginate (SA) at 0.1–0.3% (w/v) resulted in an increase in particle size and a more pronounced negative surface charge. When maintained at 4 °C over a 24-day duration, the SA-coated liposome (SA-L-2%DCPH) exhibited superior stability compared to its uncoated (L-2%DCPH) counterpart. Also, the digest derived from the SA-L-2%DCPH exhibited significantly enhanced transepithelial permeability across the Caco-2 cell monolayer, indicated by the higher protein content and ABTS radical scavenging activity. Thus, sodium alginate-coated liposomes serve as a promising delivery system for encapsulating DCPH both during storage stability and in the gastrointestinal digestion system.
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    Optimization of salt washing protocols for surimi production from invasive blackchin tilapia (Sarotherodon melanotheron): Molecular and functional properties
    (2025-10-01)
    Senphan, Theeraphol
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    Mungmueang, Natthapong
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    Hoque, Md Sazedul
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    Sriket, Chodsana
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    Benjakul, Soottawat
    This study presents the first systematic molecular characterization and protein chemistry optimization for surimi processing from invasive blackchin tilapia (Sarotherodon melanotheron), introducing novel salt washing protocols. Ten treatments were evaluated including conventional washing, single/multiple salt washing (0.3–0.9 % NaCl), and pH-shifting methods. Molecular analyses (SDS-PAGE, ATR-FTIR and SEM) revealed myofibrillar protein preservation and distinct structural modifications. The optimized 0.6 % NaCl protocol achieved selective heme protein removal while preserving myofibrillar integrity, yielding superior gel chemistry (whiteness: 76.47 vs. 71.04) and highest sensory acceptance (7.33 vs. 6.40). pH-shifting enhanced protein recovery (68.59 %) but caused molecular aggregation and network disruption, reducing gel strength by 82 %. Molecular analysis demonstrated that salt washing selectively removes chromoproteins while preserving essential gel-forming proteins, whereas pH-shifting caused protein aggregation. SEM confirmed dense protein matrices in salt-washed samples versus fragmented networks in pH-treated surimi. This chemistry-guided approach transforms invasive species into functional protein ingredients.
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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
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    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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    Impact of Bio-calcium from Nile Tilapia Bone as a Supporting Material on the Stability Enhancement of Immobilized Probiotics
    (2025-07-01)
    Petsong, Kantiya
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    Yarnpakdee, Suthasinee
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    Senphan, Theeraphol
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    Sriket, Chodsana
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    Kingwascharapong, Passakorn
    This study presented the potential of bio-calcium powder (BC) from Nile tilapia bone on enhancement of probiotics (Lactobacillus acidophilus) stability as supporting material in probiotic immobilization and novel nutritive resource, particularly calcium. Combined immobilization techniques of absorption using BC at different concentrations (0, 10, and 15%) and encapsulation using fish collagen peptide (CP) (10%) were applied. Immobilized probiotics (IP) were investigated for resistance through various adverse conditions, including freeze-drying, severe pH and temperature levels, and gastro-intestinal tract (GI) conditions. After freeze-drying, all IPs showed resistance with > 90% survivability. Under various pHs, all samples revealed non-significant differences on cell survival under neutral and alkaline conditions throughout the tested incubation period (P > 0.05). Notably decreased survivability was detected in IP with BC compared to other samples, especially under acid condition. IP with 10 and 15% BC (IP-10 and IP-15) showed higher stability to thermal condition (55 °C) with increased incubation time, compared with those without BC (IP-0). After GI digestion, IP with BC reveals a better survivability with greater recovery ability, compared with IP-0 (P < 0.05). Calcium solubility increased with BC level in a dose-dependent manner (P < 0.05). Based on morphology and microscopy analysis, IP with BC illustrated cells attached to the rough surface of BC, which was coated with collagen peptide. Elemental mappings and distribution content suggested BC could be a great supporting material and calcium resource, particularly IP-15. Therefore, the absorption by BC together with encapsulation by collagen peptide could render a highly stable immobilized probiotic and a good calcium resource.
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    Development of djenkol peel extract-loaded liposome as functional food ingredients: Physicochemical characteristic, antioxidant activities, and cytotoxicity
    (2025-06-01)
    Chotphruethipong, Lalita
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    Sinthusamran, Sirima
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    Benjakul, Soottawat
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    Senphan, Theeraphol
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    Nalinanon, Sitthipong
    Djenkol peel is an agricultural by-product containing valuable bioactive compounds. The direct use of its extract faces challenges related to stability and sensory acceptance. This study aimed to investigate the effect of djenkol peel extract (DPE) on the encapsulation efficiency (EE) using liposomes. The physicochemical properties, antioxidant activities, and cytotoxicity of the obtained liposomes were evaluated. DPE (0, 0.25, 0.50, 0.75, and 1.00 %, w/v) was encapsulated using phospholipid/cholesterol as the wall material. The highest EE (66.69 %) was observed in the liposomes loaded with 0.25 % (w/v) DPE (LE-0.25DPE) (P < 0.05). All DPE-loaded liposomes displayed nanometer-scale particle sizes (78.94–135.60 nm) and negative zeta potentials (−52.60 to −57.50 mV). Encapsulation of DPE in liposomes can mask its undesirable color. Fourier-transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), and differential scanning calorimetry (DSC) revealed changes in the physicochemical and thermal properties of LE-0.25DPE. Additionally, LE-0.25DPE showed increased DPPH radical-scavenging activity (DPPH-RSA) and ferric reducing antioxidant power (FRAP) in a dose-dependent manner (250–2000 μg/mL). In vitro Caco-2 cell viability test, LE-0.25DPE at 0.1–1 μg/mL exhibited no cytotoxicity compared to the control (P < 0.05). Thus, DPE-loaded liposomes demonstrate potential as a functional food ingredient with high bioactivity.
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    Influence of extraction methods and temperature on hemp seed oil stability: A comprehensive quality assessment
    (2025-06-01)
    Senphan, Theeraphol
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    Mungmueang, Natthaphong
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    Sriket, Chodsana
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    Sriket, Pornpimol
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    Sinthusamran, Sirima
    Hemp seed oil's increasing popularity demands a comprehensive understanding of the impact of processing on its quality and stability. This study investigated the effects of extraction methods (hydraulic pressing, Bligh and Dyer method, and Soxhlet extraction) combined with storage temperatures (45 °C and 55 °C) on oil characteristics over 60 days. The oil extraction processes were conducted using standard protocols, followed by storage under controlled conditions. Analysis encompassed color changes, oxidation indicators (peroxide value, p-anisidine value, TBARS), free fatty acids, moisture content, and Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy for molecular-level changes. Hydraulic press extraction demonstrated superior stability with minimal linoleic acid reduction (0.75 %) compared to Bligh and Dyer method (1.43 %) at 55 °C (p < 0.05) and Soxhlet extraction method (1.09 %). Higher storage temperature (55 °C) accelerated quality degradation significantly faster than 45 °C (p < 0.05), affecting oxidation markers and sensory attributes. Color darkening intensified at elevated temperatures, particularly in Bligh and Dyer extracted oils. ATR-FTIR spectroscopy analysis revealed significant changes in characteristic bands (2923.78 and 2854.07 cm⁻¹), with hydraulic press-extracted oils showing better stability in peak intensities at 45 °C compared to other methods at 55 °C (p < 0.05). While most fatty acids remained stable, polyunsaturated fatty acids showed temperature and extraction-dependent degradation. These findings highlight the critical role of extraction method selection and temperature control in preserving hemp seed oil quality, suggesting hydraulic pressing as the preferred commercial extraction method. Implementation of appropriate storage conditions and the potential incorporation of natural antioxidants could further enhance oil stability.
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    Physicochemical properties and bioavailability of bio-calcium products from tilapia bone: A comparative study with synthetic hydroxyapatite
    (2025-03-01)
    Arkanit, Kornpaka
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    Senphan, Theeraphol
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    Issapap, Nursakich
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    Mungmueang, Natthapong
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    Sriket, Pornpimol
    This study compared three calcium products derived from tilapia bones (fish bone powder, bio-calcium, and calcined bone) with synthetic hydroxyapatite as a reference material. The chemical compositions, physical properties, and calcium bioavailability were analyzed. The yield percentage decreased from 44.4 % in fish bone powder to 27.4 % in bio-calcium. The yield percentage then increased to 64.9 % in calcined bone. Synthetic hydroxyapatite showed 100 % yield. The ash content increased progressively from 68.5 % to 99.4 %. The calcium content increased from 18.9 % to 31 %, with Ca/P ratios improving from 3.3 to 1.7, approaching the ideal ratio of natural bone mineral (1.67). Heavy metals were not detected in any samples. All products exhibited high whiteness values (L∗ 91.8–95.0). Water activity values ranged from 0.33 to 0.50. The median particle sizes varied between 17.39 and 31.05 μm. Lipid oxidation decreased through the processing stages. Calcium bioavailability was the highest in synthetic hydroxyapatite (5.4 %). Bio-calcium showed 4.1 % bioavailability. The products demonstrated potential for food fortification and biomedical applications. The findings support the use of tilapia bones as a sustainable source of calcium production.