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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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    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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    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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    Comparative studies on characterizations and cytotoxicity of oil extracted from Lingzhi (Ganoderma lucidum) G2 spore using Soxhlet extraction and microwave-assisted extraction
    (2024-12-01)
    Senphan, Theeraphol
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    Benjakul, Soottawat
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    Sukketsiri, Wanida
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    Chotphruethipong, Lalita
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    This study aimed to compare the characteristics, antioxidant activity, and cytotoxicity of Ganoderma lucidum G2 spore (GLS) oil extracted using two different extraction methods, including Soxhlet extraction (SLE) and microwave-assisted extraction (MAE). The SLE method yielded higher oil content than the MAE method. MAE-extracted oil showed higher lightness (L*) and yellowness (b*) values but lower redness (a*) values. Both methods provided similar fatty acid compositions in GLS oils, including palmitic acid, oleic acid, and linoleic acid as major components. Higher saturated fatty acid content was found in SLE-extracted oil, while MAE-extracted oil had increased levels of mono- and poly-unsaturated fatty acids. The oil obtained from SLE showed slightly higher di- and tri-glycerides than that obtained from MAE (P < 0.05). Furthermore, MAE-extracted oil had high oxidative stability as indicated by lower levels of conjugated diene (CD), peroxide value (PV), thiobarbituric acid-reactive substances (TBARS), and free fatty acids (FFA) as compared to SLE (P < 0.05). MAE-extracted oil had a higher total triterpenoid content than SLE-extracted oil (P < 0.05). Also, MAE-extracted oil showed lower DPPH radical scavenging activity (DPPH-RSA) than SLE-extracted oil (P < 0.05). FTIR spectra of both samples exhibited similar functional groups. Additionally, oils obtained from both processes (at 0.1–10 µg/mL) showed low toxicity on Caco-2 cells (P > 0.05). Therefore, GLS oil with high antioxidant activity and oxidative stability, especially oil extracted by MAE, could be a food supplement for health. Also, the MAE used was a potential implement for extracting oil from GLS.
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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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    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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    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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    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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    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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    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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    A comprehensive study of diverse techniques for enhanced physicochemical and structural properties of bio-calcium from hybrid catfish bone
    (2023-12-01) ;
    Niwet, Janejira
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    Kuimalee, Surasak
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    Benjakul, Soottawat
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    Yarnpakdee, Suthasinee
    This research examined the impact of various comparative preparation methods on enhanced physicochemical, and structural properties of bio-calcium (BC) from hybrid catfish bone. Raw hybrid catfish bones (RB) were used to make bio-calcium in four steps. High-pressure water jet processing produced washed bone (WB), soaking the bone in an alkaline solution to remove proteins (PR), immersing it in ethanol to remove lipids (LR), then bleaching, and grinding it to make bio-calcium powder. The BC had greater lightness values (L*) (p < 0.05), and contained significant calcium, and phosphorus. The SDS-PAGE showed three significant protein bands with molecular weights of 250, 133, and 116 kDa. The crystalline structure of hydroxyapatite in BC was confirmed using X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy. The percentage of crystallinity of BC had a notable rise, increasing from the RB to BC of 36.25%–45.32%. The BC powder exhibited a consistent distribution of particle sizes, ranging from 2 to 10 μm. Additionally, the analysis of elemental profiles, and mappings using scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDS) revealed that BC exhibits a significant concentration of calcium, and phosphorus. BC showed superior %bioavailability of calcium (18.94%) in an in vitro simulated gastrointestinal system (GIMs) compared to RB (10.58%), and calcium carbonate (11.19%). Hence, the BC production process proved to be a viable method for acquiring premium-grade BC, which holds potential for further development as a calcium dietary supplement.
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    Development of Yellow Discoloration in Sawai (Pangasianodon hypophthalmus) Muscle due to Lipid Oxidation
    (2023-12-01) ;
    Kittiphattanabawon, Phanat
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    Patil, Umesh
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    Benjakul, Soottawat
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    Senphan, Theeraphol
    In this study, we investigated the impact of lipid oxidation on the discoloration of Sawai (Pangasianodon hypophthalmus) lipids and proteins. Sawai microsomes, liposomes, and salt-soluble myofibrillar proteins were prepared and subjected to lipid oxidation process. The results revealed that the levels of thiobarbituric acid-reactive substances, yellowness (as indicated by b* values), and pyrrole compounds increased when Sawai liposomes and microsomes were oxidized using iron and ascorbate. Meanwhile, the levels of free amines decreased, particularly as the iron content (25∼100 μM) and incubation time (0∼20 h) increased. The impact of oxidized liposomes at different levels (1, 2, and 5%) on the salt-soluble Sawai myofibrillar proteins was also evaluated. The findings revealed that lipid oxidation products reduced the sulfhydryl content and increased the surface hydrophobicity and carbonyl content of the salt-soluble Sawai myofibrillar proteins. These results imply that the formation of yellow discoloration in Sawai muscle could be due to nonenzymatic browning reactions occurring between lipid oxidation products and amines in the muscle protein.
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    Low-sodium Chaya Leaf Seasoning Powder with Potassium Chloride Substitution: Nutritional, Antioxidant, and Microbial Quality Assessment
    (2025-01-01)
    Senphan, Theeraphol
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    Puangtong, Kotchaporn
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    Namdamrassiri, Benyapa
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    Hoque, Md Sazedul
    This study investigates the development of a low-sodium seasoning powder using Chaya (Cnidoscolus chayamansa), popularly known as Mexican spinach, by substituting sodium chloride (NaCl) with potassium chloride (KCl) at various ratios. The antioxidant capabilities of dried Chaya leaves, dried at 60°C for five hours, were tested using 2,2-Diphenyl-1-picrylhydrazyl (DPPH), Ferric reducing antioxidant power (FRAP), and 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid (ABTS) techniques, demonstrating excellent free radical scavenging activity and making it suitable for low sodium seasoning compositions. The substitution of 25% KCl was the most favored option, as it effectively balanced flavor and health advantages without causing noticeable changes in appearance, color, and aroma. This recipe successfully reduced the sodium content to 1171 mg/100 g while maintaining Chaya’s original nutritional values, which include 23.57 g/100 g of protein, 4.77 g/100 g of fats, and 54.12 g/100 g of carbohydrates, as well as dietary fibers. Additionally, the formulation demonstrated physicochemical stability, as indicated by an ash content of 14.00 g/100 g, moisture content of 3.51 g/100g, and a total energy of 353.69 kcal/100 g. The safety of this new seasoning is validated by its microbiological quality, which is indicated by a total bacterial count of 2.6×10<sup>4</sup> cfu/g and the absence of mold and yeast. This emphasizes the potential of the seasoning to promote healthy dietary choices by reducing sodium intake without affecting sensory appeal.
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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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    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.