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    Evaluation of yellow mealworm (Tenebrio molitor) larvae protein extract as a cryoprotectant for frozen Pacific white shrimp
    (2026-06-15)
    Kittiphattanabawon, Phanat
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    ; ;
    Phaonakrop, Narumon
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    Roytrakul, Sittiruk
    This study evaluated the application of yellow mealworm ( Tenebrio molitor ) larvae protein extract (YMPE) as a clean-label cryoprotectant for frozen Pacific white shrimp ( Litopenaeus vannamei ), with emphasis on its underlying mechanism. Proteomic profiling identified 12 antifreeze protein-like peptides containing conserved ice-binding motifs (TCTxSxxCxxAx), supporting the cryoprotective potential of YMPE. During dose optimization, shrimp treated with 0.5% YMPE exhibited the highest weight gain (5.57%) and lowest freezable water content (59.67%) (P < 0.05). The cryoprotective efficacy of YMPE was further evaluated over five freeze–thaw cycles in comparison with distilled water and mixed phosphate. YMPE significantly mitigated quality deterioration by enhancing water retention, preserving protein functionality (higher Ca<sup>2+</sup>-ATPase activity), and reducing lysosomal enzyme leakage relative to the control (P < 0.05). Microstructural analysis revealed reduced muscle fiber disruption, characterized by fewer inter-fiber gaps. Mechanistically, YMPE may exert cryoprotective effects through a multimodal mechanism involving ice recrystallization inhibition and formation of a protective protein matrix. Although less effective than mixed phosphate, YMPE consistently improved all quality parameters compared with the control. These findings highlight the potential of insect-derived protein extracts as a novel, biologically based alternative to phosphate additives in seafood processing and provide a foundation for future industrial application.
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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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    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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    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.
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    Elucidating the physicochemical and structural properties of Ganoderma lucidum spores: Comparative analysis of various disruption techniques
    (2024-04-15) ;
    Kuimalee, Surasak
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    Yarnpakdee, Suthasinee
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    Benjakul, Soottawat
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    Sriket, Pornpimol
    This study evaluated Ganoderma lucidum G2 spores (GLS) using various processing techniques: vibrating milling, ball milling, and autoclaving. Broken GLS via vibrating and ball milling showed higher lipid content (22.89% and 22.50%) but lower L*, a*, and b* values compared to intact GLS. Vibrating milling achieved the highest sporoderm-broken rate (98.27%). Ball milling resulted in the highest lipid oxidation. Vibrating milling yielded the highest bioactive compounds, and both methods increased DPPH-RSA values. Differential scanning calorimetry indicated varying T<inf>max</inf> values: non-broken GLS (106.56 °C), vibrating milling (92.77 °C), ball milling (97.58 °C), and autoclaving (109.35 °C). FT-IR spectra showed triterpenes, polysaccharides, and fatty acids. X-ray diffraction and Fourier transform infrared spectroscopy revealed increased crystallinity with vibrating milling (37.12% to 47.99%). Both milling methods altered GLS structure to a disintegrated, pancake-like form. Scanning electron microscopy with energy-dispersive X-ray spectroscopy indicated significant carbon and oxygen concentrations. This study provided crucial insights for utilizing GLS.
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    Impact of different smoke flavors on the quality of hermetically sealed green chili paste (Nam Prik Num)
    (2024-06-01)
    Leelapongwattana, Kittima
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    Linruesee, Ratchadaporn
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    Benjakul, Soottawat
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    Yarnpakdee, Suthasinee
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    Kishimura, Hideki
    This research examined how various smoke flavors such as powdered smoke, teak, and longan wood affected the quality of hermetically sealed green chili paste (Nam Prik Num). Teak and longan wood smoking produced a darker green color than the control and smoke powder samples. The smoky aroma had no impact on yield, pH, water release, or chemical composition. However, it did influence moisture content, lightness (L*), redness (a*), yellowness (b*), and water activity (a<inf>w</inf>) compared to control and smoke-flavored samples. The attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) showed structural changes, control with C-H groups at 2924 cm<sup>−1</sup>, and smoking introduced –COOR at 1744 cm<sup>−1</sup>, signifying flavor-induced alterations. The heightened peak hinted at a distinctive smoke aroma, deepening our understanding of molecular transformations. Sample smoke flavor compounds studied using a solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS), included hydrocarbons, alcohol, aldehydes, ketones, esters, acids, and sulfide compounds, with sulfide compounds, alcohol, hydrocarbon, and alcohol compounds as the predominant volatiles. Nam Prik Num with smoke powder scored higher in appearance, color, aroma, taste, and texture than the control. Flavoring with smoke powder is more effective than charcoal grilling for adding smoky taste to Nam Prik Num.
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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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    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.