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Investigation of functional, antioxidant, anti-inflammatory, and antidiabetic properties of legume seed protein hydrolysates 豆类种子蛋白水解物的功能、抗氧化、抗炎和抗糖尿病特性研究
IF 1.9 Pub Date : 2024-01-01 DOI: 10.1016/j.fhfh.2023.100175
Taiwo Scholes Adewole , Michael Chukwuemeka Bieni , Gbenga Emmanuel Ogundepo , Oludele Olayemi Odekanyin , Adenike Kuku

Plant-derived hydrolysates are emerging as promising agents in the management of diverse ailments due to their ensuing functional and bioactive properties. This study investigated the functional, antioxidant, anti-inflammatory, and antidiabetic properties of hydrolysates from Erythrina senegalensis and Vigna subtenarrea seeds. Crude proteins were isolated via alkaline solubilization, followed by acid precipitation to the isoelectric point. Respective protein isolates were hydrolyzed using trypsin and pepsin at an enzyme-substrate ratio of 1:8 (v/v) for 1–9 h. Pepsin hydrolysates after 9 h elicited the highest solubilities of 95.54 % and 94.24 % at pH 13, while, pepsin and trypsin hydrolysates of E. senegalensis digested for 1 and 9 h displayed the highest 1,1-diphenyl-2-picrylhydrazyl radical-scavenging (10-IC50 = 2.959 mg/mL) and total antioxidant capacity (7.243 mgAAE/g), respectively. Correspondingly, pepsin and trypsin hydrolysates of E. senegalensis hydrolyzed for 5 h demonstrated the most potent anti-inflammatory activities by cogently inhibiting xanthine oxidase and lipoxygenase activities with IC50 of 0.161 ± 0.111 and 0.018 ± 0.011 mg/mL, respectively. Trypsin hydrolysates of V. subterranea hydrolyzed for 5 h potently inhibited the activities of α-amylase and α-glucosidase with respective IC50 of 0.297 ± 0.060 and 0.314 ± 0.064 mg/mL. Overall, pepsin and trypsin hydrolysates of E. senegalensis demonstrated pronounced functional bioactivities relative to V. subterranea hydrolysates. This study concludes that both seeds could serve as unique matrices of potential functional bioactive peptides with prospects for managing diabetes.

植物衍生水解物因其随之而来的功能和生物活性特性,正在成为治疗各种疾病的有前途的药物。本研究调查了塞内加尔红豆杉(Erythrina senegalensis)和亚赤藜(Vigna subtenarrea)种子水解物的功能、抗氧化、抗炎和抗糖尿病特性。通过碱性溶解分离粗蛋白,然后酸沉淀至等电点。使用胰蛋白酶和胃蛋白酶以 1:8 的酶-底物比(v/v)水解各自的蛋白质分离物 1-9 小时。同时,消化 1 和 9 小时的塞内加尔胃蛋白酶和胰蛋白酶水解物分别显示出最高的 1,1-二苯基-2-苦基肼自由基清除能力(10-IC50 = 2.959 mg/mL)和总抗氧化能力(7.243 mgAAE/g)。相应地,水解 5 小时的塞内加尔胃蛋白酶和胰蛋白酶水解物通过抑制黄嘌呤氧化酶和脂氧合酶的活性,表现出最有效的抗炎活性,IC50 分别为 0.161 ± 0.111 和 0.018 ± 0.011 mg/mL。经 5 小时水解的 V. subterranea 的胰蛋白酶水解物能有效抑制α-淀粉酶和α-葡萄糖苷酶的活性,IC50 分别为 0.297 ± 0.060 和 0.314 ± 0.064 mg/mL。总体而言,与 V. subterranea 的水解物相比,E. senegalensis 的胃蛋白酶和胰蛋白酶水解物具有明显的功能性生物活性。本研究的结论是,这两种种子都可以作为潜在功能性生物活性肽的独特基质,具有治疗糖尿病的前景。
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引用次数: 0
Corrigendum to “Peptidomic analysis of milk fermented by Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus” [Food Hydrocolloids for Health, 2021, 1, 100033] 对 "保加利亚乳杆菌亚种和嗜热链球菌发酵牛奶的肽组学分析 "的更正[《健康食品亲水胶体》,2021年,1,100033]
IF 1.9 Pub Date : 2023-12-15 DOI: 10.1016/j.fhfh.2023.100166
Yang Yu, Wenhao Yu, Yan Jin
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引用次数: 0
Advances in hydrocolloids for food applications: Natural sources, bioactivity and delivery systems 食品应用中水胶体的进展:天然来源、生物活性和输送系统
IF 1.9 Pub Date : 2023-12-07 DOI: 10.1016/j.fhfh.2023.100172
Marco Garcia-Vaquero
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引用次数: 0
Bigels for controlled gastric release of ascorbic acid: Impact on rheology, texture, thermal stability and antioxidant activity 控制胃释放抗坏血酸的凝胶:对流变学、结构、热稳定性和抗氧化活性的影响
IF 1.9 Pub Date : 2023-11-29 DOI: 10.1016/j.fhfh.2023.100171
Admajith M. Kaimal , Rekha S. Singhal

Ascorbic acid (AA) is a vital nutrient to maintain critical physiological functions but is very sensitive to processing and storage. This can be overcome by using gel-based systems for controlled release of AA. This study compares various gel-based formulations such as hydrogel, emulsion gel, bigel (25 %, 50 %, and 75 % oleogel), and emulsions for thermal stability and delivery of AA, rheological and textural profile, encapsulation efficiency (>97 %), in vitro gastrointestinal release profile, and the corresponding antioxidant profile. An increase in the oleogel content increased the hardness (125 – 216 g) and viscoelastic properties (G′ and G′′) but decreased (76.16 – 25.86 %) the swelling ratio of the bigel. A spontaneous release of AA was witnessed during gastric digestion from emulsion gels (95 %), hydrogels (98 %) and emulsions, whereas a gradual and controlled gastric release of AA could be achieved by bigels. However, a sudden decrease in AA (70 – 80 % reduction) and a spike in dehydroascorbic acid (DHA, oxidized AA) could be observed during intestinal digestion. The bioaccessibility was highest for emulsion gel and bigel (87 %) and lowest for emulsions (70 %). Bigels with higher oleogel content also showed better thermal stability but their physical stability was compromised at higher temperature. The DPPH and ABTS activity was proportional to AA, while FRAP was impacted by both DHA and AA. Thus bigels could be utilised for controlled gastric release of AA with better thermal stability.

抗坏血酸(AA)是维持人体重要生理功能的重要营养物质,但对加工和储存非常敏感。这可以通过使用凝胶为基础的系统来控制AA的释放来克服。本研究比较了各种凝胶配方,如水凝胶、乳液凝胶、凝胶(25%、50%和75%的油凝胶)和乳液对AA的热稳定性和释放、流变学和结构特征、包封效率(> 97%)、体外胃肠道释放特征和相应的抗氧化特征。随着油凝胶含量的增加,凝胶的硬度(125 ~ 216 g)和粘弹性(g′和g”)均有所提高,但凝胶的溶胀率(76.16 ~ 25.86%)有所降低。乳化液凝胶(95%)、水凝胶(98%)和乳化液在胃消化过程中自发释放AA,而凝胶可使AA在胃消化过程中逐渐和有控制地释放。然而,在肠道消化过程中,可以观察到AA突然减少(减少70 - 80%)和脱氢抗坏血酸(DHA,氧化AA)的峰值。乳凝胶和凝胶的生物可及性最高(87%),乳状液最低(70%)。油凝胶含量越高,凝胶的热稳定性越好,但温度越高,凝胶的物理稳定性越差。DPPH和ABTS活性与AA成正比,而FRAP活性受DHA和AA的共同影响。因此,bigels具有较好的热稳定性,可用于AA的胃释放控制。
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引用次数: 0
Editorial overview: VSI: Nutraceutical efficiency 编辑概述:VSI:营养保健效率
IF 1.9 Pub Date : 2023-11-28 DOI: 10.1016/j.fhfh.2023.100170
Maria Semenova (Guest Editor) , Ana Pilosof (Guest Editor)
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引用次数: 0
Therapeutic effects and mechanism of oat β-glucan plus montmorillonite powder on diarrhea in young rats 燕麦β-葡聚糖加蒙脱石粉对幼龄大鼠腹泻的治疗作用及机制
IF 1.9 Pub Date : 2023-11-08 DOI: 10.1016/j.fhfh.2023.100169
Shuyang Liu , Hamad Rafique , Liang Zou , Xinzhong Hu

Oat β-glucan is a prebiotic that affects intestinal microbiota and maintains energy homeostasis. Oat β-glucan helps treat diarrhea and other associated disorders, because it can regulate intestinal microbiota. We investigated the therapeutic effects and mechanism of oat β-glucan combined with montmorillonite powder on diarrhea induced by Senna leaf extract in young Sprague-Dawley rats. Compared with the diarrhea-induced model group (DM), the combined treatment, especially in the group treated with a medium dose of Montmorillonite powder plus oat β-glucan (M+G2), effectively reduced the diarrhea (P < 0.05) and inflammation indices, alleviated damage to the colon, and promoted weight gain in rats. In the combined treatment group, the relative abundance of Firmicutes increased at the phylum level, while the relative abundance of Proteobacteria and Actinobacteria decreased. At the genus level, the Lactobacillus content recovered, and the proportion of conditional pathogens, such as Prevotella and Paraprevotella decreased. M+G2 treatment significantly reduced diarrhea in young rats, restored intestinal microbiota diversity, and promoted the production of short-chain fatty acids (SCFAs). Based on metabolomics, the mechanism of the anti-diarrheal effect of M+G2 treatment may be related to the regulation of glucose and amino acid metabolism. The metabolic micro-environment was improved through the pentose phosphate and vitamin B6 pathways. The core metabolic regulator in metabolic network analysis was L-aspartic acid. Overall, our findings suggest that the combined treatment of oat β-glucan and montmorillonite powder may provide an effective therapeutic strategy for treating diarrhea and associated disorders by regulating the inflammatory biomarkers, SCFAs, and intestinal microbiota.

燕麦β-葡聚糖是一种影响肠道微生物群和维持能量稳态的益生元。燕麦β-葡聚糖有助于治疗腹泻和其他相关疾病,因为它可以调节肠道微生物群。研究燕麦β-葡聚糖联合蒙脱石粉对番泻叶提取物致幼年大鼠腹泻的治疗作用及机制。与腹泻模型组(DM)比较,联合给药,特别是中剂量蒙脱石粉加燕麦β-葡聚糖(M+G2)给药组,能有效减轻腹泻(P <0.05)和炎症指数,减轻结肠损伤,促进大鼠体重增加。在联合处理组中,门水平上厚壁菌门的相对丰度增加,而变形菌门和放线菌门的相对丰度降低。在属水平上,乳酸菌含量有所恢复,普雷沃氏菌和旁帕revotella等条件致病菌的比例有所下降。M+G2处理显著减少了幼龄大鼠腹泻,恢复了肠道菌群多样性,促进了短链脂肪酸(SCFAs)的产生。基于代谢组学,M+G2治疗抗腹泻作用的机制可能与调节葡萄糖和氨基酸代谢有关。通过磷酸戊糖和维生素B6途径改善了代谢微环境。代谢网络分析的核心代谢调节因子是l -天冬氨酸。总之,我们的研究结果表明,燕麦β-葡聚糖和蒙脱石粉的联合治疗可能通过调节炎症生物标志物、scfa和肠道微生物群,为治疗腹泻和相关疾病提供了有效的治疗策略。
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引用次数: 0
Exploring bioactive aloe-vera and sterculia gum to develop hydrogel wound dressings by graft-copolymerization 探索具有生物活性的芦荟和树胶接枝共聚制备水凝胶创面敷料
IF 1.9 Pub Date : 2023-11-02 DOI: 10.1016/j.fhfh.2023.100168
Baljit Singh, Jasvir Singh, Vikrant Sharma, Ankita Kumari, Diwanshi Sharma

Keeping in view the progressive future prospectus of carbohydrate research, herein, the potential of bioactive polysaccharides aloe vera (AV) and sterculia gum (SG) was explored to develop network hydrogel wound dressings. Both polysaccharides, acemannan of AV gel and glucuronic-galacturonic acid of SG accelerate wound healing process. The antibiotic drug cefuroxime encapsulated dressings were designed by graft-copolymerization with carbopol. The copolymers were characterized by SEM, AFM, FTIR, C13 NMR, XRD, TGA-DTG, DSC and swelling studies. The slow diffusion of cefuroxime drug exhibited non-Fickian mechanism and release of drug was occurred with a first order kinetic model. Polymer-bio membrane interactions indicated mucoadhesion nature. The clot formation was reduced during polymer-blood interactions indicated bio-compatible nature of the dressings moreover dressings were found antioxidant in nature. The porous dressings were O2 permeable and microbial impermeable. The hydrogel dressings revealed antimicrobial activity against P. aeruginosa. Degree of antimicrobial activity enhanced in case of hydrogel wound dressings loaded with cefuroxime. Overall, the results of these biochemical assays, sustained release of antibiotic drug and inherent wound healing potential of AV-SG indicated the use of hydrogels as dressing materials for better wound healing.

考虑到碳水化合物研究的未来前景,本文探讨了生物活性多糖芦荟(AV)和木耳胶(SG)在开发网络水凝胶伤口敷料方面的潜力。多糖、AV凝胶的葡甘露聚糖和SG的葡萄糖醛酸-半乳糖醛酸均能促进伤口愈合。采用卡波醇接枝共聚的方法设计了头孢呋辛包封敷料。通过SEM、AFM、FTIR、C13 NMR、XRD、tg - dtg、DSC和溶胀研究对共聚物进行了表征。头孢呋辛的缓慢扩散机制为非菲克机制,药物释放为一级动力学模型。聚合物-生物膜相互作用表明黏附性质。在聚合物-血液相互作用过程中,血块形成减少,表明敷料具有生物相容性,而且敷料具有抗氧化性。多孔敷料具有氧气渗透性和微生物不渗透性。水凝胶敷料对铜绿假单胞菌具有抗菌活性。在水凝胶伤口敷料中加入头孢呋辛后,抗菌活性增强。总之,这些生化试验的结果、抗生素药物的缓释和AV-SG固有的伤口愈合潜力表明,水凝胶作为敷料可以更好地愈合伤口。
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引用次数: 0
Enhanced antioxidant and anti-inflammatory effects of bee pollen and honey hybrid formulation by hot-melt extrusion 热熔挤压法增强蜂花粉与蜂蜜混合配方的抗氧化和抗炎作用
IF 1.9 Pub Date : 2023-10-22 DOI: 10.1016/j.fhfh.2023.100167
Ha Yeon Lee , Suji Ryu , Han Sol You , Yoo Na Jeon , Minki Jin , Cheong-Weon Cho , Jong-Suep Baek

Modifying the complex wall structure of bee pollen is a way to enhance the uptake of bioactive compounds. Bee Pollen and Bee Honey hybrid formulation (BPHF) are made by hot melt extrusion (HME), which is known to be the best cell wall structure-breaking method for active compound extraction because it can break the fibrous matrix of the compounds. Honey and bee pollen mixtures showed increased antioxidant activity with increased phenolic and flavonoid contents and decreased inhibitory concentration 50 (IC50) values compared to the control group. In addition, the content of gallic acid, one of the phenolic acids, increased about four times in the HME-F3 formulation compared to the control group, indicating that the phenol content was rich, but the HME-F3 formulation decreased in comparison. All samples had nano-sized particles. The Polydispersity index (PDI) value of the HME-F3 formulation was reduced and well distributed, and the stability of the particles was confirmed in the zeta potential results. Scanning electron microscopy (SEM) showed the image change of the sample after HME compared to the existing bee pollen image, and Transmission electron microscopy (TEM) showed that the particles of the mixed formulation were uniformly dispersed. Fourier-transform Infrared Analysis (FT-IR) results showed that there were structural changes in the mixture formulation compared to the control, and X-ray Diffraction Analysis (XRD) results showed that all samples were amorphous. It was found that the HME-F3 formulation could resist inflammation and protect the function of the intestinal epithelial wall. The mRNA levels of TJ-related proteins in the mixed formulation were reduced compared to LPS treatment and control.

修改蜂花粉复杂的壁结构是提高生物活性化合物吸收的一种方法。蜂花粉与蜂蜂蜜混合配方(BPHF)采用热熔挤压法(HME)制备,该方法可破坏活性化合物的纤维基质,被认为是最佳的破坏活性化合物提取细胞壁结构的方法。与对照组相比,蜂蜜和蜂花粉混合物的抗氧化活性增加,酚类和类黄酮含量增加,抑制浓度50 (IC50)值降低。此外,HME-F3配方中酚酸之一没食子酸的含量比对照组增加了约4倍,说明酚含量丰富,但HME-F3配方的酚含量相对较低。所有样品都含有纳米级颗粒。HME-F3配方的多分散性指数(PDI)值降低且分布均匀,zeta电位结果证实了颗粒的稳定性。扫描电镜(SEM)显示了HME后样品与现有蜂花粉图像的图像变化,透射电镜(TEM)显示混合配方的颗粒均匀分散。傅里叶变换红外分析(FT-IR)结果表明,与对照组相比,混合物的配方发生了结构变化,x射线衍射分析(XRD)结果表明,所有样品都是无定形的。发现HME-F3制剂具有抗炎和保护肠上皮壁功能。与LPS处理和对照组相比,混合制剂中tj相关蛋白mRNA水平降低。
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引用次数: 0
Investigating the biological activities and prebiotic potential of exopolysaccharides Produced by Lactobacillus delbrueckii and Lacticaseibacillus rhamnosus: Implications for gut microbiota modulation and rheological properties in fermented milk 研究德布吕氏乳杆菌和鼠李糖乳杆菌产生的胞外多糖的生物活性和益生元潜力:对发酵乳中肠道菌群调节和流变特性的影响
IF 1.9 Pub Date : 2023-10-16 DOI: 10.1016/j.fhfh.2023.100162
Mohammad Tarique , Abdelmoneim H. Ali , Jaleel Kizhakkayil , Ren-You Gan , Shao-Quan Liu , Afaf Kamal-Eldin , Mutamed Ayyash

The objective of this research was to explore the functional qualities of two EPSs produced by recently discovered LAB (Lactobacillus delbrueckii (EPS-LB3) and Lacticaseibacillus rhamnosus (EPS-MLB3)) that have potential probiotic benefits. The study involved evaluating their biological characteristics, such as their antioxidant, antidiabetic, antimicrobial, antibiofilm, and antiproliferative activities at various concentrations, as well as investigating their effects on the gut microbiome through in vitro fecal fermentation. Moreover, the study analyzed the rheological properties of the EPSs in fermented bovine milk. The average molecular weights of the extracted EPS were 3762.43 kDa and 1272.19 kDa with monosaccharide compositions of Glu:Rib:Man:Xyl (1.0:16.4:6.6:6.5) and Rib:Man:Xyl:GA:Ara (7.1:1.6:4.8:1.0:9.0) for EPS-LB3 and EPS-MLB3, respectively. EPS-LB3 and EPS-MLB3 at 250 mg/L showed scavenging rates of 34.0 ± 1.7 % and 37.5 ± 1.1 % for DPPH, 47.3 ± 0.8 % and 56.6 ± 0.7 % for ABTS, 38.3 ± 0.5 % and 43.5 ± 0.6 % for SD, 53.9 ± 0.1 % and 54.7 ± 0.1 % for SAS, 10.6 ± 0.1 % and 10.7 ± 0.2 % for HP, 88.8 ± 0.1 % and 84.8 ± 0.5 % for HRS, 80.0 ± 1.4 % and 84.5 ± 0.8 % for MC, as well as 60.6 ± 1.7 % and 58.1 ± 0.9 % for Lipid Oxidation, respectively, suggesting good antioxidant properties. They also exhibited antimicrobial and anti-biofilm effects against several foodborne pathogens, and antiproliferative activities against cancer cell lines. Additionally, the utilization of EPS by several probiotics indicated the prebiotic nature of EPS. The effect of both EPS on gut microbiome by fecal fermentation revealed that these EPS promoted selective bacteria like Faecalibacterium prausnitzii and Ruminococcus bromii in the gut, which are responsible for carbohydrate metabolism and short-chain fatty acid production.

本研究的目的是探索最近发现的具有潜在益生菌益处的乳酸杆菌(德氏乳杆菌(EPS-LB3)和鼠李糖乳酸杆菌(EPS-MLB3))生产的两种EPS的功能品质。这项研究包括评估它们的生物学特性,如在不同浓度下的抗氧化、抗糖尿病、抗菌、抗生物膜和抗增殖活性,以及通过体外粪便发酵研究它们对肠道微生物组的影响。此外,本研究还分析了发酵牛乳中EPSs的流变特性。提取的EPS的平均分子量分别为3762.43kDa和1272.19kDa,EPS-LB3和EPS-MLB3的单糖组成分别为Glu:Rib:Man:Xyl(1.0:16.4:6.6:6.5)和Rib:Man-Xyl:GA:Ara(7.1:1.6:4.8:1.0:9.0)。EPS-LB3和EPS-MLB3在250mg/L时对DPPH的清除率分别为34.0±1.7%和37.5±1.1%,对ABTS的清除率为47.3±0.8%和56.6±0.7%,对SD的清除率依次为38.3±0.5%和43.5±0.6%,对SAS的清除率次之为53.9±0.1%和54.7±0.1%,对HP的清除率次于10.6±0.1%和10.7±0.2%,对HRS的清除率是88.8±0.1%和84.8±0.5%,对MC的清除率则分别为80.0±1.4%和84.5±0.8%,脂质氧化分别为60.6±1.7%和58.1±0.9%,表明其具有良好的抗氧化性能。它们还表现出对几种食源性病原体的抗菌和抗生物膜作用,以及对癌症细胞系的抗增殖活性。此外,几种益生菌对EPS的利用表明了EPS的益生元性质。两种EPS通过粪便发酵对肠道微生物组的影响表明,这些EPS促进了肠道中的选择性细菌,如普氏粪杆菌和溴瘤胃球菌,它们负责碳水化合物代谢和短链脂肪酸的产生。
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引用次数: 0
Fabrication Of Butterfly Pea Flower Anthocyanin-Incorporated Colorimetric Indicator Film Based On Gelatin/Pectin For Monitoring Fish Freshness 基于明胶/果胶的蝶豌豆花花青素比色指示膜的制备及其对鱼类新鲜度的监测
IF 1.9 Pub Date : 2023-10-14 DOI: 10.1016/j.fhfh.2023.100159
Gopika Puthussery Narayanan, Preetha Radhakrishnan, Pooja Baiju, Asmath Mubeena S

Novel visual intelligent pH-indicator film was prepared from the eggshell membrane- gelatin, pectin, and anthocyanin pigment from butterfly pea flower (BP) (Clitoria ternatea). It was used as a real-time pH indicator for predicting food freshness. Eggshells are useful biowaste, gelatin has been extracted from the eggshell membrane and was used for fabricating the film. The Anthocyanin content of BP extract was 198.3 mg g−1. The film's surface morphology and chemical nature were estimated using a Scanning electron microscope, Fourier transform infrared spectroscopy, and X-ray diffraction technique. The film was pH responsive and exhibited color variation ranging from shades of red, purple, blue, green, and yellow at different pH (1-13). The applicability of the developed pH- indicator film was studied on fresh Tilapia fish by monitoring its deterioration for a specific time period. The film showed a visible color change after seven days of storage at 4ºC from dark blue, bluish-grey to olive, and deep green. Change in the total volatile basic nitrogen (TVB-N) content and pH change had an effect on the color response of the film. The findings demonstrated that the fabricated pH indicator film proved to be pH sensitive and could be used to monitor fish freshness.

以蝶豆花(Clitoria ternatea)为原料,利用蛋壳膜-明胶、果胶和花青素色素制备了一种新型的视觉智能ph指示膜。它被用作预测食品新鲜度的实时pH指标。蛋壳是一种有用的生物废弃物,从蛋壳膜中提取明胶制备了蛋壳膜。BP提取物花青素含量为198.3 mg g−1。利用扫描电子显微镜、傅里叶变换红外光谱和x射线衍射技术对膜的表面形貌和化学性质进行了估计。该膜对pH值有响应性,在不同pH值下呈现红色、紫色、蓝色、绿色和黄色等颜色变化(1-13)。通过对新鲜罗非鱼进行一定时间的变质监测,研究了所研制的pH指示膜在新鲜罗非鱼上的适用性。在4℃下保存7天后,薄膜的颜色发生了明显的变化,从深蓝色、蓝灰色到橄榄色和深绿色。总挥发性碱态氮(TVB-N)含量的变化和pH值的变化对膜的颜色响应有影响。结果表明,所制备的pH指示膜对pH值敏感,可用于鱼类新鲜度的监测。
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引用次数: 0
期刊
Food Hydrocolloids for Health
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