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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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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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A comparison of the chemical compositions and nutrient values of the sanim and srithong varieties of the farmed golden apple snails (Gastropoda; Ampullariidae; Pomacea canaliculata)
(2024-12-01)
Petcharat, Tanyamon
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Sae-leaw, Thanasak
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Benjakul, Soottawat
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Indriani, Sylvia
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Pongsetkul, Jaksuma
The comparison of the Sanim (SN) and Srithong (ST) varieties of the farmed apple snail Pomacea canaliculata as an alternative protein source has shown that ST contained more hydroxyproline and possessed a greater firmness than SN. Nevertheless, both varieties yielded similar protein contents of 20.90–23.07 % and exhibited similar texture, color, cooking yields and cooking losses after cooking. Good palatability with high liking score was obtained from both snail meat samples, particularly color, texture, and overall attributes. Snail meats possessed good nutritive value with an amount of protein and low fat content, whereas SN contained higher protein and total energy(P < 0.05). Other nutritionally useful chemicals (such as calcium, sodium, iron, vitamins B1 and B2) are present. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results discovered that myosin heavy chain, paramyosin, and actin were the prevalent protein components. Differences in nitrogen content distribution were observed between SN and ST, in which alkaline-soluble protein was the major fraction. Amino acid composition and fatty acid content varied regarding snail varieties with a large amount of essential amino acids and good source of MUFA and PUFA. Therefore, both farmed SN and ST snails could be an effective alternative protein source with a significant amount of nutritive value.
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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−1, and smoking introduced –COOR at 1744 cm−1, 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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Elucidating the physicochemical and structural properties of Ganoderma lucidum spores: Comparative analysis of various disruption techniques
(2024-04-15)
Sriket, Chodsana
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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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A comprehensive study of diverse techniques for enhanced physicochemical and structural properties of bio-calcium from hybrid catfish bone
(2023-12-01)
Sriket, Chodsana
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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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Characteristic and antioxidant activity of Cladophora glomerata ethanolic extract as affected by prior chlorophyll removal and drying methods
(2022-08-01)
Yarnpakdee, Suthasinee
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Senphan, Theeraphol
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Wangtueai, Sutee
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Jaisan, Chalalai
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Nalinanon, Sitthipong
Impact of prior chlorophyll removal (PCR) and drying methods (freeze- and oven-drying) on characteristics and antioxidant activities of the ethanolic extracts (EEs) from Cladophora (C.) glomerata, freshwater macroalgae, were investigated. The extraction yields were found in the range of 7.18%–13.15% with total phenolic (TPC) and total chlorophyll (TCC) contents of 19.40–25.69 mg GAE/g dry extract and 3.58–7.94 mg/g dry extract, respectively. Yield, TPC and TCC decreased when dechlorophyllization was implemented before extraction, regardless of drying method (p ≤.05). Increase in L* value with coincidental decrease in ΔE* value was observed in the EE with PCR. Freeze-dried EE with PCR (FEE-PCR) showed greater antioxidant efficiencies (both in vitro and oxidation model systems) than that oven-dried EE with PCR (p ≤.05). Liquid chromatography/mass spectrometry (LC/MS) analysis indicated that quercetin-3-O-glucoside was the most abundant phenolic compounds identified in the FEE-PCR. Therefore, freeze drying in combination with PCR was effective preparation method to maximize antioxidant activity and improve color of EE. Practical applications: Lipid oxidation is one of important reactions affected on the deterioration of food quality and safety. Many antioxidants have been widely used to retard lipid oxidation. Due to their health hazard of synthetic antioxidants, the search for safer ingredients with potent antioxidant activity resources is attracted interest. Phenolic compounds, the bioactive substances distributed in aquatic plants, have been known as antioxidant with health benefit. However, chlorophyll associated algal extract influenced on color and bioactivity of phenolic compound, which limits its utility. Moreover, drying technologies also have an impact on a stability of active components in algal sample. To maximize the utility of alga, the appropriate pretreatments by chlorophyll removal along with effective drying method could be a better choice for preservation of bioactive compounds. As a consequence, a wider application can be achieved. The corresponding extracts could therefore be used as an alternative natural antioxidant for food applications, instead of synthetics in a future.
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Autolysis of clown featherback (Chitala ornata) muscle
(2019-01-01)
Thiabmak, Chompunutch
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Sriket, Chodsana
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Yarnpakdee, Suthasinee
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Kim, Siriporn Riebroy
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Nalinanon, Sitthipong
Clown featherback (Chitala ornata) is a freshwater fish important in the cuisine of Thailand and other Southeast Asian countries. It has a short marketable life due mainly to poor gelling characteristic even when stored in ice. This deterioration has been associated with muscle autolysis. The research was aimed to investigate the autolysis of clown featherback muscle (CFM). To test autolysis, mince from CFM was incubated in a temperature controlled water bath for 60 min at 5, 10, 20, 30, 40, 50, 60, 65, 70, 75 or 80 °C at pH levels over the range of 2-11. The results showed that the highest autolytic activity was at 70 °C and it occurred at pH levels of 4 and 7, with pH 4 showing the highest autolysis. CFM was incubated with its crude enzyme counterpart at the physiological pH (6.73) at both 4 °C and 25 °C. Degradation of natural actomyosin showed that after incubation at either temperature myosin heavy chain was susceptible to hydrolysis indicating that autolysis of CFM took place even at low temperature as affected by its endogenous proteases. The effects of several protease inhibitors were tested and autolysis at the physiological pH (6.73) was strongly inhibited by 10 μM pepstatin A (20.3% inhibition), which had significantly higher inhibitory activity (P < 0.05) among the protease inhibitors tested. This suggests that aspartic protease was the major proteinase in CFM.
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Physicochemical characteristics of glucosamine from blue swimming crab (Portunus pelagicus) shell prepared by acid hydrolysis
(2018-12-01)
Kraisangsri, Jittrakan
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Nalinanon, Sitthipong
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Riebroy, Siriporn
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Yarnpakdee, Suthasinee
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Ganesan, Palanivel
The aim of this research was to characterize glucosamine hydrochloride (GluHCl) from the shell of blue swimming crab (Portunus pelagicus). The crab shell was finely milled and processed to chitin prior to HCl hydrolysis using 30 % HCl for 30 min at 100 ºC for glucosamine production. The resultant glucosamine was recovered by crystallization using 95 % ethanol and was dried in a hot air oven. The color of the glucosamine crystals, expressed as L*, a*, and b*, was 83.01, 5.03, and-3.38, respectively. Crab shell glucosamine had high purity, which could be strongly stained by ninhydrin and presented at the same Rf of standard D-glucosamine using thin layer chromatography. Furthermore, prepared glucosamine exhibited similar Fourier transform infrared (FTIR) spectrum as standard D-glucosamine. Glucosamine from blue swimming crab shell had high purity as determined by HPLC and contained 808.15 mg D-glucosamine/g sample. The maximal transition temperature (T<inf>max</inf>) and the total enthalpy (ΔH) of prepared glucosamine were 194 ºC and 754.42 J/g, respectively. As a consequence, with the presented method, the resultant glucosamine was characterized to be D-glucosamine. Therefore, blue swimming crab shell, a byproduct from crab meat processing, has high potential as a raw material to produce glucosamine for food and nutraceutical applications.
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Lipid oxidation and fishy odour development in protein hydrolysate from Nile tilapia (Oreochromis niloticus) muscle as affected by freshness and antioxidants
(2012-06-15)
Yarnpakdee, Suthasinee
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Benjakul, Soottawat
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Nalinanon, Sitthipong
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Kristinsson, Hordur G.
Lipid oxidation and fishy odour development in protein hydrolysate from fresh and ice-stored Nile tilapia (Oreochromis niloticus) were investigated. During iced storage of 18 days, heme iron content decreased with a concomitant increase in non-heme iron content (P < 0.05). Peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) values increased. Phospholipid content decreased with a corresponding increase in free fatty acid content. The results suggested that lipid hydrolysis and oxidation took place during storage. When protein hydrolysates were produced from fresh and 18 days ice-stored Nile tilapia muscle, higher lipid oxidation and fishy odour/flavour along with higher amount volatile compounds were obtained in hydrolysate for unfresh sample (P < 0.05). However, the addition of mixed antioxidants during hydrolysis process markedly lowered lipid oxidation, b ·, ΔC ·, ΔE · values, fishy odour/flavour as well as the formation of volatile compounds in the resulting hydrolysates prepared from both fresh and unfresh samples. Therefore, hydrolysate from Nile tilapia muscle with reduced fishy odour and lighter colour could be prepared by using fresh fish and incorporation of mixed antioxidants during hydrolysis. © 2011 Elsevier Ltd. All rights reserved.

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