دانشگاه تبریزپژوهش های صنایع غذایی2008-515X36120260522Comparison of Astaxanthin Pigment Extraction from Shrimp Shells and Gammarus Crustacean Using Microwave and Soaking in Ionic Liquid Microemulsion in Waterمقایسه استخراج رنگدانه آستاگزانتین از پوست میگو و سختپوست گاماروس به کمک مایکروویو و خیساندن در میکروامولسیون مایع یونی در آب1132181410.22034/fr.2026.62708.1940FAپریسافیضیدانشکده صنایع غذایی دانشگاه علوم کشاورزی و منابع طبیعی گرگانیحییمقصودلوگروه علوم و مهندسی صنایع غذایی دانشکده صنایع غذایی دانشگاه علوم کشاورزی و منابع طبیعی گرگانهدیشهیری طبرستانیگروه شیمی مواد غذایی، دانشکده صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگانسید مهدیجعفریگروه علوم و مهندسی صنایع غذایی دانشکده صنایع غذایی دانشگاه علوم کشاورزی و منابع طبیعی گرگانامیر هوشنگبحریگروه علوم شیلات مرکز تحقیقات فناوری های دریایی و شیلاتی واحد بندر عباس دانشگاه آزاداسلامی بندرعباسJournal Article20240729Background: The diversity and anticancer properties of carotenoid pigments have attracted much attention. Carotenoids are yellow and red pigments found in bacteria, fungi, plants, and animals (Fidor and Borda, 2014). Carotenoids inactivate the initiators of harmful chemical reactions, such as free radicals. On the other hand, due to their strong antioxidant properties, they prevent the oxidation of unsaturated fatty acids and have shown important functions in the pharmaceutical, food, and cosmetic industries (Sachindra and Mahendrakar, 2005). Many studies have been conducted on the extraction of carotenoid pigments from crustaceans such as crabs and shrimps (Parjiklai, Al-Houri, Ferte, & Christensen, 2015; Soumya & Sachindra, 2015), of which astaxanthin and its esters are the most abundant (Sachindra, Bhaskar, & Mahendrakar, 2006). Among the important shrimp species, banana shrimp is the second most important commercial species found in Hormozgan waters. Among the crustaceans, Gammarus is an apodous crustacean and an important commercial species in some countries and has a high concentration of carotenoids, protein, various types of enzymes, and essential unsaturated fatty acids. This crustacean plays an important role in cleaning the aquatic environment of fish such as salmon, and their presence in the fish diet increases the rate of digestion and absorption of food, resulting in increased growth performance (Escobar-Lux, Parsons, Samuelson, & Agnalt, 2020). In recent studies, this Atlantic crustacean has been studied and investigated as a rich source of carotenoid pigments (Namati, Shokri, & Pazouki, 2015). There are various methods for extracting pigments, including chemical methods (Hooshmand, Shabanpour, Mousavinasab, & Golmakani, 2017), microbial methods (Das et al., 2007), and the use of enzymatic digestion by proteolytic enzymes (N-Sachindra & Mahendrakar, 2011). Each of the mentioned extraction methods has advantages and disadvantages. The conventional soaking method for the extraction of carotenoid pigments is usually time-consuming and involves high cost and large volumes of solvent. Also, in most cases, these methods have lower efficiency compared to new methods such as microwave-assisted extraction and ultrasound (Li, Fabiano-Tixir, Tomao, Krautow, & Chamet, 2013). Microwave-assisted extraction is based on the absorption of microwave energy by polar molecules of chemical compounds (Rotary and Orsat, 2012). In recent years, in order to reduce the effects of volatile and toxic solvents, hydrophilic ionic liquids have been used as dispersing solvents and hydrophobic ionic liquids as extracting solvents. Ionic liquids are a salt in liquid form that is considered among green solvents due to its tunable physicochemical properties, high chemical and thermal stability, and negligible vapor pressure at room temperature, and has a molecular structure consisting of different cations and anions (Khu et al., 2019). However, the viscosity of most ionic liquids is higher than that of organic solvents, which results in a decrease in the mass transfer rate. Microemulsion is a promising method that enables selective extraction of biomolecules in the food and chemical industries (Amiri-Rigi and Abbasi, 2019). Ionic liquid microemulsion provides a suitable environment for the release and extraction of astaxanthin due to its adjustable polarity, low surface tension and ability to solubilize hydrophobic compounds. Also, its high stability and ability to penetrate the biological matrix increase the extraction efficiency compared to traditional solvents. To date, microemulsions have been used to extract proteins, pigments and trace elements. Since a high percentage of aquatic waste consists of wastes that contain many value-added compounds, their extraction not only contributes to the economic prosperity of the fisheries industry but also contributes significantly to environmental protection. Gao et al. (2020) extracted astaxanthin from shrimp waste using ultrasound and ionic liquid microemulsion. The microemulsion containing tributyl octylphosphonium bromide significantly increased the extraction of astaxanthin due to stronger electrostatic interactions and hydrogen bonding (Gao et al., 2020). In this regard, Nunes et al. (2021) extracted astaxanthin as a carotenoid with high antioxidant capacity from crab exoskeleton waste for use in food products. This extraction involved a microwave pretreatment step (with hydroalcoholic solvents with 0-50% water by volume at different temperatures from 40 to 140°C) and supercritical fluid extraction (at a pressure of 200-500 bar, a temperature of 40-60°C, and an ethanol content of 8-13% by weight). The extracted astaxanthin content was reported to be 12 times higher than that of the traditional Soxhlet extraction method, indicating that this proposed method significantly improves the extraction efficiency (Nunez et al., 2021).<br /><br />Objective: The aim of this study was to compare astaxanthin extraction from shrimp (Fenneropenaeus merguiensis) and gammarus crustacean (Pontogammarus maeoticus) using soaking and microwave-assisted methods in the presence of ionic liquid microemulsion.<br /><br />Methods: To extract astaxanthin from lyophilized powder of banana and hard shell shrimp (Gammarus spp.) skin, the traditional method of soaking in a microemulsion solvent of ionic liquid in water (5:1, v/v) at ambient temperature for 24 hours was used. Microwave extraction was also performed under pre-optimized conditions with a frequency of 2.45 GHz in a 5-fold solvent to sample ratio, a power of 100 W, and a time of 91.81 seconds (Faizi et al., 2025). After extraction, the obtained extract was diluted with ethanol and then filtered with a 0.45 μm syringe filter to prepare for analysis (Fan et al., 2019). Next, in order to compare the extraction of astaxanthin using two methods, soaking and microwave, tests were used for the amount of total carotenoids, astaxanthin, recovery percentage, and antioxidant properties.<br /><br />Results: According to the results, the highest yield of astaxanthin was obtained from shrimp (80.39 ± 1.09 µg/ml) using the microwave method. Therefore, shrimp was identified as a better source of astaxanthin compared to gammarus. Furthermore, the recovery percentage and total carotenoid content for shrimp were 93% and 59%, and 83.54 ± 0.56 ml/g and 77.98 ± 1.33 ml/g for the microwave and soaking methods, respectively, indicating the superiority of the microwave-assisted extraction. In addition, the antioxidant activity of astaxanthin extracted by the soaking method was higher than that of the microwave method. Compared to the synthetic antioxidant BHT, the antioxidant activity of astaxanthin was always greater, and increased with concentration.<br /><br />Conclusion: Crustacea waste can be used as the cheapest raw material for the extraction of carotenoid pigments. On the other hand, considering that the usual methods of carotenoid extraction are time-consuming and require a lot of solvent, the use of modern extraction methods such as microwave has become common today. Overall, based on the findings of this study, shrimp waste can be considered a suitable and effective source for astaxanthin extraction using microwave-assisted methods.زمینه مطالعاتی: ضایعات سختپوستان میتوانند بهعنوان ارزانترین مواد اولیه جهت استخراج رنگدانههای کاروتنوئیدی استفاده شوند. از طرف دیگر باتوجه به اینکه روشهای معمول استخراج کارتنوئیدها زمانبر بوده و نیاز به حلال زیادی دارند امروزه استفاده از روشهای نوین استخراج مانند مایکروویو رایج شده است. <br />هدف: این پژوهش با هدف مقایسه استخراج آستاگزانتین از پوست میگویموزی (Fenneropenaeus merguiensis) و سختپوست گاماروس (Pontogammarus maeoticus) به کمک دو روش خیساندن و مایکروویو انجام شد. <br />روش کار: پس از تهیه پودر نمونهها و میکروامولسیون مایع یونی در آب، فرایند استخراج با نسبت 5 برابر حلال به نمونه، به کمک روش خیساندن در دمای اتاق به مدت 24 ساعت و روش مایکروویو با فرکانس 45/2 گیگاهرتز، توان100 وات و زمان 81/91 ثانیه انجام شد. <br />نتایج: طبق یافتهها بیشترین میزان استخراج آستاگزانتین از میگوی موزی 09/1± 39/80 میکروگرم بر میلیلیتر به کمک روش مایکروویو بدست آمد. طوریکه میگوی موزی نسبت به سختپوست گاماروس منبع بهتری برای استخراج آستاگزانتین بود. علاوه براین درصد بازیافت و مقدار کاروتنوئید کل برای میگوی موزی با استفاده از روش مایکروویو و خیساندن به ترتیب 93 و 59 درصد، 56/0±54/83 و 33/1±98/77 میلی لیتر بر گرم بود که نشان دهنده برتری روش مایکروویو نسبت به روش خیساندن است. همچنین فعالیت آنتیاکسیدانی آستاگزانتین استخراج شده از روش سنتی بالاتر از روش مایکروویو بود و در مقایسه با آنتیاکسیدان سنتزیBHT، با افزایش غلظت همواره فعالیت آنتی اکسیدانی در هر دو روش پایینتر از BHT بود.<br />نتیجه گیری: بهطور کلی طبق یافتههای پژوهش حاضر ضایعات میگوی موزی منبع مناسبی برای استخراج رنگدانه آستاگزانتین میباشد.https://foodresearch.tabrizu.ac.ir/article_21814_927b64f94accf3419145bb2c921a7425.pdfدانشگاه تبریزپژوهش های صنایع غذایی2008-515X36120260522Impact of Tryptophan Treatment on Maintaining Marketability and Extending the Shelf Life of Sweet Cherry Fruitتأثیر تیمار تریپتوفان بر حفظ بازارپسندی و افزایش عمر انبارمانی میوه گیلاس15322181510.22034/fr.2026.70769.1984FAنرمینکریمیاندانشجوی دکتری علوم و مهندسی باغبانی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایرانفرهنگرضویاستاد، گروه علوم و مهندسی باغبانی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایرانمرتضیسلیمانی اقدمدانشیار، گروه علوم باغبانی، دانشکده کشاورزی و منابع طبیعی، دانشگاه بینالمللی امام خمینی قزوین، قزوین، ایرانJournal Article20251217Introduction<br />Sweet cherry (Prunus avium L.) is a popular stone fruit globally, prized for its desirable flavor, high nutritional value, and significant antioxidant properties (Nava-Ochoa et al., 2025). As a non-climacteric fruit with low ethylene production during ripening, it is highly perishable and has a short postharvest life (Blando and Oomah, 2019). This fruit faces substantial postharvest challenges due to its high respiration rate and susceptibility to fungal decay, leading to rapid quality losses such as reduced firmness, stem browning, and undesirable changes in color, flavor, and nutritional value during storage (Mujtaba et al., 2023). In recent years, the use of bioactive compounds to elicit defense responses has emerged as a sustainable postharvest strategy. Amino acids (AAs), as organic compounds and protein precursors, act as biostimulants that enhance plant metabolic efficiency and growth (Matysiak et al., 2020; Trovato et al., 2021). By scavenging reactive oxygen species (ROS) and maintaining cellular integrity, they can reinforce plant defense systems against environmental stresses. Among them, tryptophan (β-indolyl alanine), an aromatic amino acid biosynthesized via the shikimate pathway, has found extensive applications in food, medical, and agricultural industries (Xiao et al., 2023). In the postharvest context, exogenous application or endogenous accumulation of tryptophan has been linked to reduced chilling injury and fungal decay, delayed senescence, and quality preservation in fruits. These beneficial effects are largely attributed to its role as a precursor for key bioactive molecules such as melatonin, serotonin, auxin, and nicotinamide (NAD+), which are involved in signaling regulation and strengthening plant defense mechanisms (Yuxiao et al., 2023; Aghdam and Arnao, 2024). Supporting evidence highlights the efficacy of tryptophan in various fruits. For example, a 100 µM tryptophan treatment in strawberries and blueberries increased endogenous melatonin and salicylic acid accumulation while reducing ethylene and abscisic acid production, thereby extending storability. Furthermore, by supplying necessary nitrogen for cell wall synthesis, tryptophan enhanced firmness in cherries and raspberries by reinforcing pectin, hemicellulose, and lignin structures (Arabia et al., 2025). Previous studies indicate that postharvest tryptophan application can preserve fruit quality by upregulating phenolic biosynthesis genes (e.g., CHS and PAL), increasing NAD+/NADH and NADPH/NADP+ ratios, and activating the antioxidant system (e.g., enhancing SOD, CAT, and APX gene expression), thereby reducing ROS accumulation, as demonstrated in strawberry (Zhou et al., 2024). In ‘Le-Conte’ pear, preharvest tryptophan treatment (100 ppm) improved fruit set, yield, weight, skin color (L* and a*), total soluble solids (TSS), total soluble carbohydrates, total phenols, and total amino acid content (Khedr, 2018), and subsequently helped maintain TSS and reduce quality loss during 12 weeks of cold storage (Khedr, 2019). Despite promising results in various fruits, information regarding the impact of tryptophan on the physicochemical and marketability attributes of sweet cherry remains limited. Consequently, this study was designed to evaluate the effect of different concentrations of postharvest tryptophan application on maintaining quality, delaying senescence, extending shelf life, and enhancing the antioxidant system of ‘Tak Daneh’ sweet cherries during cold storage.<br /><br />Materials and Methods<br />Sweet cherry fruits (Prunus avium L. cv. ‘Tak Daneh’) were harvested at commercial maturity on June 21, 2023, from an orchard in Saqqez, Kurdistan Province, Iran, and promptly transported to the Postharvest Physiology Laboratory, Department of Horticultural Sciences, University of Zanjan. Fruits with uniform size and color and free of visible defects were immersed for 20 minutes in tryptophan solutions at 0 (control), 0.5, 1, and 2 mM (Sigma Chemical Co.) containing 0.01% Tween-20 as a surfactant. The experiment was conducted as a factorial arrangement based on a completely randomized design (CRD) with three replications per treatment combination. After air-drying, fruits were placed in polyethylene containers and stored at 1 ± 0.5 °C and 85-90% relative humidity. A comprehensive set of postharvest quality and biochemical attributes was evaluated after 0, 7, 14, 21, and 28 days of storage. For assessments on days 7-28, fruits were equilibrated at room temperature (20±1 °C) for 24 h before analysis to simulate market conditions. Day-0 measurements were performed on three freshly harvested, untreated fruits. The key quality parameters were assessed using established methods as follows: the stem browning index (SBI) was determined using a visual scoring method (Yang et al., 2019); stem chlorophyll content was measured spectrophotometrically according to Arnon (1967); total anthocyanin content (TAC) was quantified using the pH-differential method (Giusti and Wrolstad, 2001). Fruit color coordinates (L*, a*, b*, C*, h°, ΔE) were obtained using a digital colorimeter (Lutron RGB-1002) with calculations based on standard formulas (Shahabi-Ghahafarrokhi et al., 2015; Mujtaba et al., 2023). Titratable acidity (TA), total soluble solids (TSS), and pH were analyzed from fruit juice, and the flavor index was calculated as the TSS/TA ratio (Naser et al., 2018). Total soluble carbohydrates (TSC) were assayed using the anthrone method (Irigoyen et al., 1992). Proline content was determined via the ninhydrin assay (Sánchez et al., 2001). Total soluble protein (TSP) was quantified using the Bradford method (Bradford, 1976). Peroxidase (POD) activity was measured by monitoring guaiacol oxidation (Zhang et al., 2013). Total antioxidant capacity was evaluated based on DPPH radical scavenging activity (Dehghan and Khoshkam, 2012). Data were analyzed using SPSS (version 26) software. Analysis of variance (ANOVA) was performed, and mean separation was carried out using Duncan’s multiple range test at a significance level of p < 0.05. Graphs were prepared with Microsoft Excel 2019.<br /><br />Results and Discussion<br />Analysis of variance (ANOVA) indicated that tryptophan treatment, storage duration, and their interaction significantly affected the stem browning index and most colorimetric and biochemical traits, including L*, a*, C*, hue angle, ΔE, anthocyanin content (TAC), stem chlorophyll (TChl), pH, titratable acidity (TA), total soluble solids (TSS), total soluble carbohydrates (TSC), proline (Prol), total soluble protein (TSP), peroxidase (POD) activity, and total antioxidant capacity (p < 0.01). For the b* coordinate, the main effect of tryptophan treatment was not significant, whereas both storage time and the treatment × storage interaction had a highly significant effect (p < 0.01). Moreover, the flavor index was significantly influenced by the interaction between tryptophan application and storage duration (p < 0.05). Consistent with these statistical findings, postharvest application of tryptophan, particularly at 2 mM, markedly strengthened the antioxidant defense system and alleviated oxidative damage in ‘Tak Daneh’ sweet cherries during cold storage. Fruits treated with 2 mM tryptophan exhibited higher levels of proline, total soluble protein, peroxidase activity, total antioxidant capacity, and total soluble anthocyanins, indicating improved metabolic adaptation to storage-induced stress. These biochemical adjustments contributed to the preservation of key physicochemical and sensory attributes, including pH, titratable acidity, total soluble solids, flavor index, total soluble carbohydrates, and a significantly reduced incidence of stem browning. From a mechanistic perspective, the observed enhancement in antioxidant capacity and anthocyanin content can be attributed to tryptophan's role as a precursor for key signaling molecules. Tryptophan can enhance endogenous melatonin synthesis by upregulating genes such as TDC, T5H, SNAT, and ASMT (Madebo et al., 2021; Sharafi et al., 2021). Melatonin, a potent indoleamine with anti-senescence properties, subsequently stimulates the accumulation of phenolics, flavonoids, and anthocyanins by activating phenylalanine ammonia-lyase and chalcone synthase while suppressing polyphenol oxidase (Sharafi et al., 2021; Magri and Petriccione, 2022). Furthermore, tryptophan may elevate anthocyanin levels by upregulating key biosynthesis genes like DFR and UFGT (Miranda et al., 2020; Zhou and Zhang, 2024). Concurrently, tryptophan is implicated in stimulating endogenous salicylic acid (SA) production (Arabia et al., 2025). SA enhances plant tissue resistance by altering fruit physiology and boosting secondary metabolites, which likely contributed to delayed senescence and reduced stem browning in treated fruits. Thus, the combined action of tryptophan-derived melatonin and SA, along with a direct boost to the enzymatic antioxidant system (e.g., POD activity), effectively mitigated oxidative stress, preserving the overall quality and extending the marketable life of sweet cherries during cold storage. Therefore, the superior preservation of quality attributes in tryptophan-treated 'Tak Daneh' cherries, as evidenced by the biochemical and physical data, can be mechanistically linked to this multifaceted, elicitor-induced defense response.<br /><br />Conclusion<br />Postharvest application of tryptophan at 2 mM is an effective, natural strategy for maintaining quality, extending shelf life, and enhancing the antioxidant defense system in sweet cherry during cold storage. This treatment mitigates oxidative damage by elevating proline, soluble proteins, peroxidase activity, and antioxidant and anthocyanin contents, thereby improving biochemical resilience and marketability of ‘Tak Daneh’ sweet cherry fruit.تریپتوفان بهعنوان یک اسیدآمینه آروماتیک و پیشساز ترکیبات زیستفعال، میتواند در تنظیم پاسخهای دفاعی و تأخیر در فرآیند پیری نقش مؤثری ایفا کند. میوه گیلاس (Prunus avium L.) در دوره پس از برداشت، بهعلت داشتن پوست نازک، سرعت تنفس زیاد و فعال شدن مسیرهای اکسیداتیو، فسادپذیری بالایی دارد. بنابراین، استفاده از تریپتوفان میتواند رویکردی مناسب برای بهبود ماندگاری آن بهشمار آید. این پژوهش با هدف ارزیابی تأثیر تیمار پس از برداشت تریپتوفان بر ویژگیهای بیوشیمیایی و آنتیاکسیدانی در میوه گیلاس رقم «تکدانه» در شرایط انبار سرد انجام شد. آزمایش به صورت فاکتوریل در قالب طرح کاملاً تصادفی با سه تکرار اجرا گردید. فاکتور اول شامل غلظتهای مختلف تریپتوفان (صفر، 5/0، 1 و 2 میلیمولار) و فاکتور دوم مدت زمان انبارمانی (7، 14، 21 و 28 روز) بود. میوهها پس از غوطهوری در تیمار، در دمای 5/0±1 درجه سانتیگراد و رطوبت نسبی 85 تا 90 درصد نگهداری شدند. نتایج نشان داد، که کاربرد تریپتوفان، بهویژه در غلظت 2 میلیمولار، بهطور معنیداری موجب افزایش پرولین، پروتئین محلول کل، فعالیت آنزیم پراکسیداز، ظرفیت جاروب کنندگی رادیکال DPPH و آنتوسیانین کل شد. این تغییرات با حفظ اسیدیته و اسید کل، کنترل افزایش مواد جامد محلول و کربوهیدرات محلول کل، بهبود شاخص طعم و نیز کاهش شدت قهوهای شدن دم میوه در طی دوره انبارمانی همراه بود. بهطور کلی، یافتهها نشان داد، که تریپتوفان میتواند با تقویت سیستم آنتیاکسیدانی و کاهش آسیب اکسیداتیو، بهعنوان یک ماده زیستفعال و طبیعی برای حفظ کیفیت و افزایش ماندگاری پس از برداشت میوه گیلاس در شرایط انبار سرد مورد استفاده قرار گیرد.https://foodresearch.tabrizu.ac.ir/article_21815_24f82ed93ff0eb4ad6d248f3285439c6.pdfدانشگاه تبریزپژوهش های صنایع غذایی2008-515X36120260522Comparison of the effects of inulin and maltodextrin on the Pickering emulsions properties stabilized using whey protein microgels via the Maillard reactionمقایسه تأثیر اینولین و مالتودکسترین بر ویژگیهای امولسیونهای پیکرینگ پایدارشده با میکروژل پروتئین آبپنیر از طریق واکنش میلارد33482181610.22034/fr.2024.64480.1950FAسید حسینایزدیگروه علوم و مهندسی صنایع غذایی- دانشکده کشاورزی- دانشگاه تهرانغلامرضاعسگریگروه علوم و صنایع غذایی، دانشکده کشاورزی، دانشگاه تهرانزهراامام جمعهگروه علوم و مهندسی صنایع غذایی، دانشکده کشاورزی دانشگاه تهرانمریمسلامیگروه علوم و صنایع غذایی، دانشکده کشاورزی- دانشگاه تهرانفاطمهقدرتیگروه علوم و صنایع غذایی- دانشکده کشاورزی- دانشگاه تهرانJournal Article20241113Introduction: Pickering emulsions, an innovative emulsion system, have recently attracted significant attention. These emulsions achieve stability through the use of solid particles rather than synthetic surfactants, offering high resistance to coalescence. Unlike surfactants, these particles are insoluble in water and oil, and they stabilize the emulsion by forming a physical barrier at the oil-water interface, which prevents droplet merging. Such features make Pickering emulsions a sustainable, environmentally friendly, and safe choice for applications in the food, pharmaceutical, and cosmetic industries. With the rising demand among consumers for natural and safer products, there is increasing interest in developing biocompatible, particle-stabilized Pickering emulsions. In recent years, biopolymers—often referred to as “green” alternatives—have been explored to replace synthetic emulsifiers in food products. Biopolymers, including proteins and carbohydrates, offer nutritional value, natural origins, and biocompatibility, making them appealing as replacements for industrial emulsifiers. Proteins are useful for emulsification due to their amphiphilic nature but can be unstable under external processing conditions such as temperature, pH, and salts. Carbohydrates, on the other hand, enhance emulsion stability by increasing viscosity but are not surface-active enough to function as emulsifiers on their own. However, combining and modifying proteins and saccharides can enhance their emulsifying properties. Key modifications include forming protein microgel structures or conjugating proteins with carbohydrates via the Maillard reaction, which improves their functional stability, thermal resistance, and solubility. Whey protein, a cheese industry byproduct rich in essential amino acids, is known for its rapid absorption and high bioavailability. Due to its favorable functional properties, including three-dimensional network formation, strong particle bonding, and biocompatibility, whey protein microgels are considered a promising stabilizer for Pickering emulsions. Conjugating whey protein microgels with saccharides such as inulin and maltodextrin via the Maillard reaction can further enhance their emulsifying capabilities, affecting properties like surface tension, and improving stability against temperature and pH fluctuations. Inulin and maltodextrin are commonly used saccharides in food science. Maltodextrin, a tasteless, water-soluble oligosaccharide, is widely used due to its low cost and ease of production from corn starch. In contrast, inulin, a fructan polysaccharide with prebiotic benefits, is non-digestible and reaches the large intestine intact. It is valued for its ability to reduce calorie intake, lower blood lipids, and increase satiety, making it popular in low-calorie foods as a thickening and stabilizing agent. This study aims to investigate the effectiveness of whey protein microgel conjugates with inulin and maltodextrin in stabilizing Pickering emulsions. The primary focus is to assess the influence of saccharide type and the Maillard reaction on emulsion stability, structure, and physicochemical properties in systems with a high internal phase. The findings contribute to understanding polysaccharide roles in protein conjugates, supporting the development of more stable and safer Pickering emulsions for various industries.<br />Materials and Methods: Whey protein isolate was dissolved in Milli-Q water to a concentration of 40 mg/mL. The protein dispersion was stirred at 25 °C for 2 hours, then incubated at 4 °C for 12 hours to ensure full hydration. The pH was adjusted to 5.80 ± 0.05 using 1.0 M HCl with a pH-meter. Next, the dispersion was heated in a water bath at 85 °C for 15 minutes, without stirring, to form whey protein microgels (WPMs), and rapidly cooled to 4 °C in an ice-water bath. Freshly prepared WPM was then combined with inulin (In) and maltodextrin (Md) at a 2:2 protein-to-saccharide ratio, and these were labeled as In:WPM and Md:WPM, respectively. Additionally, a mixture containing both saccharides in a 2:1:1 protein:inulin:maltodextrin ratio was prepared and labeled In:Md:WPM. All mixtures were prepared at a total concentration of 4% w/w. The pH was adjusted to 8.0 using NaOH. The dispersions were then heated in a 90 °C water bath for 1 hour, followed by rapid cooling in an ice-water bath. Samples were dialyzed against Milli-Q water (pH-matched) using a 14,000 Da molecular weight cutoff membrane, and stored at 4 °C for further analysis. WPM heated without carbohydrate was labeled WPM. To ensure repeatability, three separate samples were prepared per run, with each experiment conducted in triplicate. After the Maillard reaction, degree of grafting (DG), particle size, zeta potential, and surface tension of the nanoparticles were measured. For HIPPE preparation, soybean oil (φoil = 0.80) was added to both conjugated and unconjugated WPM dispersions (pH = 7.0). Emulsification was achieved via homogenization at 24,000 rpm for 4 minutes using an Ultra-Turrax disperser, and the emulsions were analyzed on the same day. FTIR spectroscopy, droplet size, polydispersity index (PDI), emulsification activity index (EAI), emulsification stability index (ESI), creaming index, and viscosity of HIPPEs were also determined.<br />Results and Discussion: Regarding modification/conjugation degree, the Maillard reaction enabled covalent bonding between whey protein microgel and saccharides, with inulin (polysaccharide) showing a higher degree of conjugation than maltodextrin (oligosaccharide) due to its longer chain length and greater availability of carbonyl groups. Without carbohydrates, modification in WPM was minimal, likely from trace lactose interactions. The particle size increased with saccharide addition, with inulin contributing more significantly due to its larger chain, which expanded the WPM particles’ volume. This larger size supports stability in high internal-phase Pickering emulsions. After the Maillard reaction, increased zeta potential improved electrostatic repulsion and stability, as all samples carried a negative charge at pH 7 (above WPM’s isoelectric point). All conjugated WPM particles reduced surface tension, a critical factor for emulsion stability. To assess the emulsification performance and interfacial properties of emulsifiers, surface tension is an important parameter, as it is closely related to the stabilization of emulsions against coalescence. The adsorption mechanism of emulsifiers includes two steps: first, the emulsifiers diffuse and attach to newly formed oil droplets, resulting in a steep drop in surface tension. In the second stage, the emulsifiers rearrange at the interface, causing a slight decrease in surface tension until it levels off. Inulin showed a greater reduction in surface tension than maltodextrin due to its long, unbranched chain, which better covers the particle surfaces. In terms of HIPPEs, FTIR analysis indicated characteristic protein-related amide bands and triglyceride peaks from the oil phase, with changes in amide peaks supporting protein modification during glycation. Moreover, saccharide addition led to smaller oil droplets, which are more stable, as larger WPM particles occupied more space, restricting oil mobility and droplet size. WPM conjugated with carbohydrates, especially inulin, showed higher emulsion activity index than unconjugated WPM. Inulin's large and linear structure enabled better surface protection around oil droplets, enhancing stability. Similarly, conjugated WPM-stabilized emulsions had greater stability than unconjugated ones. Inulin's structure formed a stronger protective layer compared to maltodextrin, reducing droplet agglomeration and enhancing stability. Also, conjugation with saccharides, particularly inulin, increased viscosity and emulsion uniformity. The rheological properties of Pickering emulsions are influenced by two key factors: the viscosity of the continuous phase and the structural characteristics of the particles, such as their size, shape, and rigidity. Higher viscosity supports stability by increasing the interfacial area and promoting shear-thinning behavior. Due to the formation of products with higher molecular mass, hydrophilicity, surface charge, and steric hindrance , HIPPEs created using inulin-conjugated WPMs exhibited a higher consistency coefficient (K) and greater resistance to applied shear stress. Finally, the creaming index decreased with carbohydrate addition, particularly with a higher inulin-to-maltodextrin ratio, due to increased oil droplet density and viscosity, reflecting the positive impact of the Maillard reaction on stability and creaminess.<br />Conclusion: The findings of this study indicate that conjugating whey protein microgels (WPM) with inulin and maltodextrin via the Maillard reaction is an effective method for producing stable Pickering emulsions with a high internal phase. This covalent binding with polyols significantly enhances the surface activity of WPM, allowing it to be adsorbed at the oil-water interface and play a crucial role in stabilizing the emulsions. Among the compounds tested, WPM conjugates with inulin exhibited a greater impact on improving emulsion properties and stability. This enhanced functionality is likely due to inulin’s longer molecular chains, which provide stronger steric repulsion, lower surface tension, and form more stable interfacial layers compared to maltodextrin. Overall, this research highlights the substantial potential of the Maillard reaction for enhancing emulsion performance and boosting the emulsifying capacity of natural proteins by conjugating them with saccharides. Conjugated whey protein nanoparticles with inulin and maltodextrin show promise as an effective approach for creating stable Pickering emulsions with favorable physicochemical characteristics for applications in the food, pharmaceutical, and cosmetic industries. Additionally, these Pickering emulsions can serve as precursors or integral components in innovative products, such as oleogels, paving the way for further advancements in these fields.در این پژوهش به منظور ساخت امولسیون پیکرینگ با محتوای فاز پراکنده بالا از میکروژل حاصل از ایزوله پروتئین آبپنیر اصلاحشده از طریق انجام واکنش میلارد استفاده شد. برای این منظور، در مرحله نخست میکروژل پروتئین آب پنیر تولید و در مرحله بعد با اینولین، مالتودکسترین، و ترکیب اینولین و مالتودکسترین با نسبت مساوی،از طریق انجام واکنش میلارد به صورت کووالانسی متصل شد. محصولات مزدوج شده حاصل در کنار میکروژل پروتئین آبپنیر مزدوج نشده، به عنوان نمونه فاقد پلیساکارید یا نمونه شاهد، در تولید امولسیون پیکرینگ استفاده شده و توانایی امولسیفایریو پایدار کنندگی آنها همراه با خصوصیات امولسیونهای بدست آمده بررسی و مقایسه شدند. امولسیون پایدارشده با نانومزدوج میکروژل- اینولین یک شبکه متراکم با کوچکترین اندازه قطرات روغن 8.9 میکرومتر و کمترین اندیس خامهایشدن (34%) را به وجود آورد. همچنین، این امولسیون بیشترین میزان ویسکوزیته، فعالیت امولسیونشوندگی، و پایداری فیزیکی را نشان داد. این دستآورد مربوط به افرایش درجه مزدوجشدن و پتانسیل زتا، و همینطور کاهش کشش سطحی نانوذرات میکروژل اصلاحشده با پلیساکارید در مقایسه با نمونه میکروژل فاقد کربوهیدرات بود. طیفسنجی FTIR به منظور بررسی گروه های عملکردی اجزای مختلف امولسیونهای پیکرینگ با فاز داخلی بالا انجام شد. با توجه به مقادیر پایین پروتئین و پلیساکارید استفادهشده و علیرغم اینکه اینولین و مالتودکسترین هیچکدام ژلساز نیستند و توانایی ایجاد شبکه ندارند، واکنش میلارد در ترکیبات مزدوجشده توانست نقش موثری در تولید امولسیون پیکرینگ با فاز پراکنده بالا ایفا کند.https://foodresearch.tabrizu.ac.ir/article_21816_81719f524c9720f54a5f15e03690897e.pdfدانشگاه تبریزپژوهش های صنایع غذایی2008-515X36120260522Effect of bioactive compounds on oxidative and lipolytic stability in cow's milk-based functional drinkتاثیر ترکیبات زیستفعال بر پایداری اکسیداتیو و لیپولیتیکی نوشیدنی فراسودمند بر پایه شیر گاو49602091110.22034/fr.2025.66650.1962FAبیوک آقافرمانیگروه علوم و صنایع غذایی، دانشکده کشاورزی و منابع طبیعی اهر، دانشگاه تبریز، ایرانصمدبدبدکگروه علوم و صنایع غذایی، دانشکده کشاورزی و منابع طبیعی اهر، دانشگاه تبریز، ایرانفرشتهسلمانیگروه علوم و صنایع غذایی، دانشکده کشاورزی، دانشگاه تربیت مدرس، ایرانJournal Article20250407Introduction: Lipid peroxidation often occurs in response to oxidative stress and reactive oxygen species cause oxidation of lipids containing (C=C) carbon double bonds (Sies and Jones 2020). Malondialdehyde (MDA), the most common aldehyde with the highest biological activity, is abundantly produced during lipid peroxidation and is commonly used as an indicator of oxidative stress (Barrera et al. 2018). MDA originates from the breakdown of peroxide compounds resulting from the oxidation of polyunsaturated fatty acids (Ayala et al. 2014). Carrots contain natural, nutritious and health-promoting bioactive compounds. The antioxidant properties of β-carotene and other compounds such as phenolic compounds, flavonoids and ascorbic acid in carrots are of great interest (Anjani et al. 2022). The aim of this study was to investigate the effect of different percentages of carrot juice in the preparation of carrot-milk drink. Therefore, during the thirteen-day storage period, quality indicators such as acid number, MDA, antioxidant capacity and bioactive compounds were studied in the drinks to determine the appropriate percentage of carrot juice to inhibit oxidative stress in cow’s milk based functional drink.<br />Materials and Methods: Carrots and pasteurized cow's milk were obtained from local market. The carrots were washed with clean water, extracted and passed through a filter cloth to separate the carrot pieces. Milk-based drinks were prepared by replacing 0 (only cow milk) 10, 20, 30, 40 and 50% of carrot juice. The samples were filled into sealed glass containers and pasteurized at 70°C for 30 min and stored at 4°C until the experiments. Malondialdehyde and acid number tests were performed on the milk-carrot drink with methanol and isoamyl alcohol (lipolysis index), as well as bioactive compounds such as total phenolic compounds, total carotenoids, ascorbic acid and antioxidant capacity in the drink samples. Malondialdehyde measurement: The thiobarbituric acid method was used to measure malondialdehyde in beverage samples and its amount was reported as µmol MDA L-1 (Fenaille et al. 2001). Acid number measurement (lipolysis index): To determine the total free fatty acids, the method of titration of extracted fat by an alkaline alcoholic solution was used and its amount was expressed in mg NaOH g-1 fat (Evers 2003). Determination of bioactive compounds: 1) Total phenol measurement: Methanol-water solvent (50:50) was used to extract the phenolic compounds of the beverage and the Folin-Ciocalteau method was used by measuring the absorbance of the samples at 765 nm to determine the amount of phenolic compounds. Finally, the amount of phenolic compounds was reported as mg GAE 100 g-1 (Lamuela‐Raventós 2018), 20 Measurement of total carotenoids: Hexane-ethanol solvent (90:10) was used to extract the carotenoid compounds of the beverage and centrifugation (5000 rpm) was used for purification. The amount of total carotenoids (μg g-1) was determined by measuring the absorbance of the sample at 450 nm (Machmudah and Goto 2013), 3) Measurement of ascorbic acid: Ascorbic acid was extracted using a metaphosphoric acid-acetic acid solvent and its amount was determined by measuring the absorbance at 521 nm and the amount of ascorbic acid was reported as mg 100 g-1 (Ruiz et al. 2016) and 4) Measurement of total antioxidant capacity by DPPH test: The antioxidant capacity of the sample was determined by the free radical scavenging ability of the DPPH solution. For this purpose, the absorbance at 517 nm was determined and reported as percentage inhibition (Chen et al. 2015). For statistical analysis, factorial experiments were carried out in a completely randomized design with 3 replications. The effect of carrot juice percentage and storage time on bioactive compounds and oxidative and lipolytic stability of carrot-milk drink was investigated. SAS software (version 9.1, USA) was used for analysis and the least mean squares method (P < 0.05) was used for comparison of means.<br />Results and discussion: Samples with 50% carrot juice on the first day and 10% carrot juice on the thirteenth day had the highest (157.37 µg g-1) and lowest (24.44 µg g-1) carotenoid contents, respectively. The decrease in total carotenoid content during storage can be attributed to the antioxidant and free radical scavenging properties of carrot-milk drink, as carotenoids exhibit antioxidant properties and are capable of scavenging reactive oxygen species at low oxygen concentrations (Ribeiro et al. 2018). Carotenoids have the ability to bind to lipid globules and provide phase and color stability during storage in carrot-milk drink (Sharma et al. 2012). Studies have shown that carrot juice contains significant amounts of vitamins such as thiamine, folic acid, niacin, riboflavin, vitamins A and C (Aubert et al. 2022). During storage, vitamin C decreased in all samples due to degradation and oxidation (Castellom-Estrada et al. 2023). The content of ascorbic acid decreased during storage. The lowest content (2.15 mg 100g-1) on the thirteenth day with 10% carrot juice and the highest content (6.17 mg 100g-1) was observed with 50% carrot juice on the first day. In all samples, the lowest content of total phenols (122.18 mg 100g-1) was on the thirteenth day with 10% carrot juice and the highest content (311.32 mg 100g-1) on the fourth day with 50% carrot juice. The major source of phenolic compounds in carrot-milk drink is carrot juice. The increase in phenolic compounds up to the seventh day can be attributed to the release of phenolic compounds from carrot particles into the drink, but the subsequent decrease is due to oxidation inhibition, antimicrobial properties and preservative role during storage (Talcott and Howard 1999). The lowest antioxidant capacity was found in the sample containing 10% carrot juice on the first day (35.98%) and the highest value was found in the sample containing 50% carrot juice on the thirteenth day (60.62%). Researches has shown that there is a good correlation between DPPH free radical scavenging values, phenolic compounds and vitamin C content on the one hand, and antioxidant capacity and beta-carotene on the other (Castellom-Estrada et al. 2023 and Hashemi et al. 2014). Acid number, as an indicator of lipase enzyme activity, was lower in drinks with 30, 40 and 50% carrot juice than in drinks with 10 and 20% carrot juice, and these changes in acid number were calculated as 0.5-1.2 mg NaOH g-1 in terms of extracted fat. Also, the researchers found that the presence of Pseudomonas spp. in milk leads to the production of microbial lipases at the end of storage. According to the results of the experiments, the decrease in the acid value can be attributed to the inhibitory effect of bioactive compounds from the carrot juice source, especially at higher percentages in the carrot-milk drink. The results of the experiments showed that the MDA content of the drinks changed from 3.5 to 13 µmol L-1 during storage. As a result, the most important pathways for inhibiting and suppressing lipid peroxidation and MDA formation could be the inhibition of free radicals such as hydroxyl (OH·) and peroxyl (LOO·) radicals with the help of bioactive compounds in carrot juice. Aldehydes are the major secondary oxidation products in milk, which generally have a lower taste threshold compared to alcohols and ketones. As a result, when their concentration reaches above the taste threshold, they have a significant effect on the taste of milk. Also, MDA can damage biomolecules such as proteins, phospholipids and other molecules through the formation of covalent and cross-links (Aubourg 1993). Researchers reported that the addition of purple corn anthocyanin to milk resulted in higher resistance to lipid oxidation and MDA formation, and showed lower oxidation of secondary metabolites compared to the control sample (Tian et al. 2022). In lipid oxidation, autoxidation is the most important chain reaction through free radicals. Investigation of Table 1 shows that milk-carrot drinks contain significant amounts of bioactive compounds (polyphenolic compounds, flavonoids, carotenoids, vitamin C) and antioxidant capacity that have the ability to inhibit the peroxidation of unsaturated fatty acids and the formation of MDA (Figures 1 and 2). Lipid peroxidation, in addition to the loss of nutritional value, also leads to the formation of radicals with toxic properties (Figure 3), therefore, the control of oxidative processes in the food industry have a vital importance. Although the oxidation of unsaturated fatty acids has been widely studied, the complex reactions involved in this process, as well as the different pathways and factors affecting them, have caused the oxidation mechanisms to be not yet fully understood (Dominguez et al. 2019). Unsaturated fatty acids are very sensitive to oxidant attack and today, malondialdehyde, 4-hydroxy-2-nonenal, and 2-isoprostane are the main biomarkers for assessing lipid peroxidation, all of which are derived from polyunsaturated fatty acids (Mas-Bargues 2021).<br />Conclusion: Carrots, as a rich source of carotenoid compounds such as α and β-carotene, different types of phenolic compounds and ascorbic acid, have antioxidant and free radical neutralization properties in carrot-milk drink. The main source of phenolic compounds in carrot-milk drink is carrot juice. During whole storage, changes in antioxidant compounds of carrot-milk drink may be due to inhibition of the formation of harmful compounds such as MDA resulting from the secondary oxidation of unsaturated fatty acids. The most important pathways for inhibiting and suppressing lipid peroxidation and MDA formation can be the inhibition of free radicals such as hydroxyl and peroxyl radicals with the help of bioactive compounds in carrot juice. These compounds can also act as lipolysis inhibitors during storage. Finally, it can be concluded that adding carrot juice with different concentrations to carrot-milk drink, in addition to increasing nutritional value, also plays an important role in the oxidative and lipolytic stability of functional drink.زمینه مطالعاتی: گونههای فعال اکسیژن باعث اکسیداسیون لیپیدهای حاوی پیوندهای دوگانه کربن-کربن میشود. مالون-دیآلدئید (MDA) متداولترین آلدئید با بالاترین فعالیت بیولوژیکی، به طور فراوان طی پراکسیداسیون لیپیدی تولید می-گردد و معمولاً بهعنوان شاخص تنش اکسیداتیو استفاده میشود. <br />هدف: هویج دارای انواع ترکیبات زیستفعال طبیعی و مغذی با خواص آنتیاکسیدانی است که برای سرکوب تنش اکسیداتیوی و MDA مورد توجه میباشد.<br />روش کار: نوشیدنیهای بر پایه شیر با جایگزینی 0 (فقط شیر گاو)، 10، 20، 30، 40 و 50% آبهویج تهیه و در ظروف شیشهای دربسته پر و در دمای C◦ 70 برای min 30 پاستوریزه و تا انجام آزمایشات در دمای °C 4 نگهداری شدند. آزمونهای MDA، عدد اسیدی، ترکیبات فنولی، کاروتنوئید کل، آسکوربیک اسید و ظرفیت آنتیاکسیدانی در نمونهها انجام شد.<br />نتایج: عدد اسیدی بهعنوان شاخص تعیین فعالیت آنزیم لیپاز، در نوشیدنیهای با 30، 40 و 50% آب هویج کمتر از نوشیدنیهای با 10 و 20% آب هویج مشاهده شد که این تغییرات عدد اسیدی mg NaOH g-1 2/5-1/0 بر حسب چربی استخراجی محاسبه شد. نتایج آزمایشها نشان داد محتوای MDA نوشیدنیها از 5/3 بهµmol L-1 13 طی نگهداری تغییر کرده است. در نتیجه مهمترین مسیرهای مهار و سرکوب پراکسیداسیون لیپیدها و تشکیل MDA، میتواند مهار رادیکالهای آزاد مانند رادیکال هیدروکسیل (OH·) و پراکسیل (LOO·) با کمک ترکیبات زیستفعال آب هویج باشند.<br />نتیجهگیری: هویج با منبع غنی ترکیبات کاروتنوئیدی، پلیفنولها و آسکوربیک اسید و خواص آنتیاکسیداسیونی برای خنثیسازی رادیکالهای آزاد در نوشیدنی شیر-هویج طی ذخیرهسازی باعث مهار تشکیل ترکیبات مضر مانند MDA حاصل از اکسیداسیون ثانویه اسیدهای چرب غیراشباع شد.https://foodresearch.tabrizu.ac.ir/article_20911_10ac996881c0a49f9ab515f938624ee4.pdfدانشگاه تبریزپژوهش های صنایع غذایی2008-515X36120260522Effect of replacing wheat flour with pumpkin powder on the batter viscosity and the physicochemical and sensory properties of pancakesاثر جایگزینی آرد گندم با پودر کدوحلوایی بر ویسکوزیته خمیر و خصوصیات فیزیکوشیمیایی و حسی پنکیک61782158510.22034/fr.2026.70712.1983FAفخرالدینصالحیاستاد، گروه علوم و صنایع غذایی، دانشکده صنایع غذایی، دانشگاه بوعلی سینا، همدان، ایرانسپیدهوجدانی وحیددانشجوی کارشناسی ارشد، گروه علوم و صنایع غذایی، دانشکده صنایع غذایی، دانشگاه بوعلی سینا، همدان، ایرانJournal Article20251213Introduction: The food industry currently faces the dual challenge of meeting the nutritional needs of a growing population while responding to consumer demand for functional foods. Bakery products, as widely consumed foods, provide an ideal platform for nutritional enrichment. Utilizing unconventional, bioactive-rich ingredients such as fruit and vegetable powders offers an innovative strategy to enhance health benefits and diversify products (Abedfar & Sadeghi, 2016; Salehi & Aghajanzadeh, 2020; Sarmasti et al., 2023; Adesunmibo et al., 2025).<br />Pumpkin (Cucurbita moschata) is rich in carotenoids (especially beta-carotene), phenolic compounds, vitamins, minerals, and dietary fiber, making it a promising ingredient for food enrichment. These compounds not only provide antioxidant activity but also influence sensory and physicochemical properties of the final product (Rakcejeva et al., 2011; Hosseini Ghaboos et al., 2018). Previous studies have shown that adding pumpkin powder can significantly increase moisture, ash, and mineral content in cakes, while improving sensory attributes and consumer acceptance (Jalali et al., 2018; Hosseini Ghaboos et al., 2018; Khormali et al., 2021).<br />Pancakes, as a popular bakery product, can serve as a suitable carrier for these bioactive compounds (Chen et al., 2022; Vejdanivahid & Salehi, 2025). However, partial substitution of wheat flour with pumpkin powder introduces technical challenges, affecting dough rheology, water balance, starch–protein interactions, and gluten network formation. These changes, in turn, influence physicochemical properties such as color, texture, moisture, and bioactive content, which ultimately affect key sensory attributes, including appearance, aroma, taste, texture, and overall acceptability (Salehi & Aghajanzadeh, 2020).<br />Despite scattered studies on pumpkin powder in bakery products, there is a lack of comprehensive research simultaneously evaluating the effects of increasing substitution levels on dough rheology, final product physicochemical properties, and sensory acceptance. This study aims to fill this gap by investigating the impact of replacing wheat flour with 0, 10, 20, 30, and 40% pumpkin powder on pancake dough rheology, physicochemical properties (color, texture, phenolic content, antioxidant activity), and sensory characteristics (appearance, aroma, taste, texture, overall acceptability). It is hypothesized that pumpkin powder will enhance nutritional and antioxidant properties, induce measurable changes in dough and product properties, and, at optimal levels, produce pancakes with high sensory acceptability, paving the way for industrial production of enriched, healthy, and consumer-friendly bakery products.<br />Material and methods:<br />Materials: Wheat flour (Zar Macaroni, Iran), vanilla (Hamishk, Iran), baking powder (Myofl, Iran), sugar (Zamen, Iran), full-fat pasteurized milk (3.4% fat, Damdaran, Iran), fresh eggs (Telavang, Iran), sunflower oil (Tabiat, Iran), and fresh pumpkin were sourced locally in Hamadan, Iran. Chemical reagents included Folin–Ciocalteu reagent, sodium carbonate, gallic acid, DPPH, and ethanol.<br />Pumpkin Powder Preparation: Fresh pumpkin was washed, sliced thinly, dried at 70°C for 100 min, and ground into powder using an industrial mill.<br />Pancake Preparation: An optimized formulation was used, with gradual substitution of wheat flour with pumpkin powder at 0, 10, 20, 30, and 40%. Egg whites were whipped to form a stable foam, yolks were emulsified with oil, and dry ingredients were gradually incorporated. The batter rested for 10 min at room temperature before cooking (25 g per pancake at 180-190 °C).<br />Batter viscosity: In this study, after preparing the pumpkin powder, the pancake batter was formulated according to an optimized recipe, and the different substitution levels were applied. Apparent viscosity of pancake batter measured using a rotational viscometer at 5, 10, and 15 rpm.<br />Physicochemical properties: Moisture, ash, pH, titratable acidity, baking loss, volume, density, phenolic content (Folin–Ciocalteu), and antioxidant activity (DPPH assay).<br />Color and texture: Color parameters obtained via image analysis (RGB to L*, a*, b*), crust hardness assessed by penetration test.<br />Sensory evaluation: Conducted with 20 trained panelists for appearance, aroma, taste, texture, and overall acceptability.<br />Statistical Analysis: All experiments were conducted in triplicate. Data were analyzed using one-way ANOVA, and mean comparisons were performed with Duncan’s multiple range test at p<0.05.<br />Results and discussion:<br />Batter viscosity: The addition of pumpkin powder significantly increased the apparent viscosity of pancake batter at all shear rates (5, 10, and 15 rpm). The effect was dose-dependent, with the 40% substitution showing the highest viscosity. This increase is attributed to the high water-holding capacity and fibrous nature of pumpkin powder, which strengthens the batter network. The batter exhibited pseudoplastic behavior over 120 s, with higher substitution levels showing greater rheological stability.<br />Color parameters: Pumpkin powder significantly affected batter color, decreasing lightness (L*) and increasing redness (a*) and yellowness (b*), reflecting the natural carotenoid content. Lightness decreased from 89.1 (control) to 81.2 (40%), redness increased, and yellowness rose from 32.5 to 71.2. These changes may influence consumer perception, as color is a key quality indicator.<br />Physical properties: Increased pumpkin powder led to higher pancake weight (23.2 g to 24.5 g), reduced baking loss (7.2% to 1.2%), increased volume (20 cm³ to 23.7 cm³), and decreased density (1158 kg/m³ to 1031 kg/m³). These results indicate better moisture retention, lighter texture, and improved structural properties.<br />Moisture, ash, pH, and acidity: Moisture and ash content increased significantly with higher substitution, enhancing nutritional value. pH decreased (8.15 to 6.10) while acidity increased (0.30% to 0.63%), reflecting the presence of natural acids and phenolic compounds in pumpkin powder.<br />Phenolic content and antioxidant activity: Total phenolic content and antioxidant capacity rose with substitution, from 967.1 to 1659.8 µgGAE/g and 35.4% to 79.3%, respectively, demonstrating pumpkin powder as a rich source of bioactive compounds.<br />Crust hardness: Pancakes became progressively softer with higher pumpkin levels, likely due to the formation of a hydrocolloidal matrix and reduced gluten content.<br />Sensory evaluation: While appearance scores decreased due to darker color, aroma, taste, texture, and overall acceptability improved with higher pumpkin levels. The 40% substitution achieved the highest overall acceptance, indicating strong potential for consumer-preferred fortified products.<br />Conclusion: Partial replacement of wheat flour with pumpkin powder effectively enhances the nutritional and functional properties of pancakes. Increasing substitution up to 40% improved batter viscosity, product volume, moisture, and texture, while significantly raising total phenolic content and antioxidant activity. Although darker color slightly reduced visual appeal, aroma, taste, and texture improved, resulting in the highest overall acceptability for the 40% sample. These findings indicate that pumpkin powder can produce health-promoting pancakes with desirable texture and strong consumer acceptance.زمینه مطالعاتی: محصولات نانوایی مانند پنکیک، بهعنوان بستری مناسب برای غنیسازی، میتوانند با استفاده از منابع غیرمتعارف مانند پودر میوهها و سبزیها، ارزش غذایی و سلامتبخش خود را افزایش دهند. کدوحلوایی با دارا بودن ترکیبات ارزشمندی نظیر کاروتنوئیدها، فنلها و فیبر، گزینهای امیدوارکننده برای این منظور محسوب میشود.<br />هدف: هدف اصلی این مطالعه، ارزیابی تأثیر جایگزینی آرد گندم با پودر کدوحلوایی در سطوح صفر (شاهد)، ۱۰، ۲۰، ۳۰ و ۴۰ درصد بر ویژگیهای رئولوژیکی خمیر پنکیک، ویژگیهای فیزیکوشیمیایی و ویژگیهای حسی محصول نهایی بود.<br />روش کار: ویسکوزیته خمیر، شاخص رنگ، وزن، افت پخت، حجم، چگالی، رطوبت، خاکستر، pH، اسیدیته، میزان فنل کل، فعالیت آنتیاکسیدانی (DPPH)، سختی پوسته و ویژگیهای حسی (ظاهر، عطر، طعم، بافت و پذیرش کلی) با روشهای استاندارد ارزیابی و دادهها با استفاده از آنالیز واریانس یکطرفه تجزیه و تحلیل شدند.<br />نتایج: نتایج نشان داد که افزودن پودر کدوحلوایی تا 40 درصد ویسکوزیته خمیر را بهطور معنیداری (05/0>p) افزایش و سختی پوسته محصول پختهشده را بهصورت معنیداری (05/0>p) کاهش داد. همچنین باعث افزایش معنادار (05/0>p) وزن، حجم، رطوبت، خاکستر، میزان فنل کل و فعالیت آنتیاکسیدانی و کاهش معنادار (05/0>p) افت پخت، چگالی، pH و روشنایی رنگ شد. در ارزیابی حسی، با وجود کاهش پذیرش ظاهر به دلیل تیرگی رنگ، امتیاز عطر، طعم، بافت و پذیرش کلی بهویژه در نمونه حاوی ۴۰ درصد، بهبود یافت و این نمونه بالاترین امتیاز پذیرش کلی را کسب کرد.<br />نتیجهگیری نهایی: در مجموع میتوان بیان کرد که پودر کدوحلوایی تا سطح ۴۰ درصد، پتانسیل بالایی برای تولید پنکیک غنیشده با ترکیبات زیستفعال و بهبودیافته از نظر ویژگیهای بافتی و حسی دارد.دانشگاه تبریزپژوهش های صنایع غذایی2008-515X36120260522Investigation of Encapsulation Stability and Antioxidant Properties of Lutein in Iota-Carrageenan-Inulin Hydrogelبررسی پایداری درونپوشانی و خواص آنتیاکسیدانی لوتئین در هیدروژل یوتا-کاراگینان-اینولین79912181710.22034/fr.2026.72136.1990FAاکرمپزشکیگروه علوم و صنایع غذایی -دانشکده کشاورزی- دانشگاه تبریزتبریز-ایران'گلارااسدیگروه علوم و مهندسی صنایع غذایی -دانشکده کشاورزی -دانشگاه تبریز-تبریز-ایرانبابکقنبرزادهگروه علوم و مهندسی صنایع غذایی-دانشکده کشاورزی-دانشگاه تبریز-تبریز-ایرانJournal Article20260428Introduction<br />Lutein is a lipophilic carotenoid belonging to the xanthophyll family, abundantly found in dark green leafy vegetables (e.g., spinach, kale), egg yolk, and certain fruits. Due to its polyene structure with conjugated double bonds, lutein exhibits potent antioxidant activity and plays a critical physiological role in protecting the macula lutea of the retina from blue light-induced oxidative damage. Epidemiological and clinical studies have consistently demonstrated that adequate dietary intake of lutein reduces the risk of age-related macular degeneration (AMD) and cataracts, two leading causes of blindness worldwide. Beyond ocular health, lutein also contributes to systemic antioxidant defense, skin photoprotection, and cognitive health. However, the commercial application of lutein in functional foods, pharmaceuticals, and cosmetics is severely hampered by its high sensitivity to light, heat, oxygen, and acidic pH, as well as its poor water solubility and low bioavailability. Conventional surfactant-based delivery systems (e.g., Tween 80 micelles) fail to provide adequate physical barriers against pro-oxidant agents, leading to rapid degradation and loss of bioactivity. Therefore, there is an urgent need to develop advanced encapsulation systems that can protect lutein, enhance its stability, and enable controlled release. In this context, polysaccharide-based hydrogels have emerged as promising carriers due to their biocompatibility, biodegradability, and ability to form three-dimensional hydrophilic networks capable of entrapping lipophilic bioactives after appropriate stabilization.<br /><br />Iota-carrageenan (ι-carrageenan) is a sulfated linear polysaccharide extracted from red seaweeds (Rhodophyta). In the presence of divalent cations such as Ca²⁺, ι-carrageenan undergoes a conformational transition from random coils to helical structures, followed by aggregation into a three-dimensional elastic, transparent, and thermally reversible gel network. Despite its excellent gelling properties, pure ι-carrageenan hydrogels suffer from limited mechanical strength, high syneresis, and insufficient barrier properties against oxygen diffusion—limitations that restrict their use in long-term protection of highly oxidizable compounds. Inulin, a natural fructan-type soluble dietary fiber derived from chicory roots, Jerusalem artichoke, garlic, and onion, offers a complementary functionality. In addition to its well-established prebiotic effects (stimulating beneficial gut microbiota such as Bifidobacteria and Lactobacilli), inulin increases total solid content, promotes the formation of microcrystalline domains upon cooling, and enhances gel network density. When combined with ι-carrageenan, inulin is expected to produce a hybrid hydrogel with improved compactness, reduced oxygen permeability, and enhanced encapsulation efficiency for lutein. Despite extensive research on carrageenan or inulin individually, no systematic study has yet investigated the ι-carrageenan-inulin hybrid hydrogel as a carrier for lutein. Therefore, the present study aimed to: (1) fabricate ι-carrageenan-inulin hybrid hydrogels containing three different inulin concentrations (15, 20, and 25% w/w); (2) characterize the structural, morphological, and physicochemical properties of the hydrogels using X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM); (3) evaluate the encapsulation efficiency (EE%) and storage stability (30 days) of lutein in the hydrogel systems compared to a non-encapsulated control (lutein in Tween 80); and (4) assess the concentration-dependent antioxidant activity of lutein using the DPPH radical scavenging assay and determine the IC50 value.<br /><br />Materials and Methods:<br />Iota-carrageenan (Sigma-Aldrich, Germany), inulin (≥90% purity from chicory, Sigma-Aldrich, USA), lutein (≥90%, Extrasynthese, France), Tween 80, n-hexane, absolute ethanol, calcium chloride (CaCl₂), and DPPH (2,2-diphenyl-1-picrylhydrazyl) were purchased from Merck (Germany). Hydrogels were prepared by dissolving 3% (w/w) ι-carrageenan in deionized water at 70°C under magnetic stirring (500 rpm, 30 min). Inulin was added at 15%, 20%, or 25% (w/w) and stirring continued until complete dissolution and transparency. The solution was cooled to 50°C, and CaCl₂ was added to a final concentration of 20 mM to induce gelation. For lutein loading, 0.1 g lutein was dispersed in 4 g Tween 80 (30 min stirring at 25°C), then incorporated into the polymer solution prior to CaCl₂ addition. The mixtures were stored at 4°C for 24 h to complete gelation. A control sample (lutein in Tween 80 without hydrogel) was also prepared. For EE% determination, 1 g of hydrogel was mixed with 9 mL n-hexane, vortexed (2 min), centrifuged (4000 rpm, 10 min, 4°C), and the free lutein in the supernatant was quantified spectrophotometrically at 441 nm. Stability was evaluated over 30 days at 25°C under ambient light by measuring residual lutein at 5-day intervals. XRD patterns were recorded using a Siemens D500 diffractometer (Cu-Kα radiation, λ=0.154 nm, 40 kV, 30 mA, 2θ=5-70°, step size 0.05°, scan rate 1°/min). FTIR spectra were obtained on a Bruker Tensor 27 spectrometer (KBr pellets, 400-4000 cm⁻¹, resolution 4 cm⁻¹). SEM micrographs were taken on a TESCAN MIRA3 FEG-SEM after gold sputtering. DPPH assay was performed by mixing different lutein concentrations (10–50 mg/mL in ethanol) with 0.1 mM DPPH solution (1:3 v/v), incubating in darkness for 30 min, and measuring absorbance at 517 nm. Percentage inhibition was calculated, and IC50 was determined by linear regression. All experiments were performed in triplicate. Statistical analysis was conducted using SPSS version 26 (one-way ANOVA followed by Tukey’s post hoc test, significance level p < 0.05).<br />Results and Discussion:<br />Encapsulation Efficiency (EE%): Increasing inulin concentration from 15% to 25% significantly improved EE%, with values of 71.2 ± 1.2%, 76.5 ± 1.5%, and 82.1 ± 1.2% for 15%, 20%, and 25% inulin, respectively (p < 0.05). This enhancement is attributed to increased viscosity of the continuous phase, greater polymer chain entanglement, and formation of a denser gel network with reduced mesh size at higher inulin levels, which physically entraps lutein more effectively and limits its diffusion out of the matrix.<br />XRD Analysis: The XRD pattern of the control hydrogel (without lutein, H) exhibited a broad halo between 2θ = 18–25°, characteristic of predominantly amorphous polysaccharide matrices with limited semi-crystalline domains originating from local chain ordering through hydrogen bonding. After lutein incorporation (H+L sample), the diffraction intensity decreased significantly and the peak broadened, indicating a reduction in crystallinity and an increase in the amorphous fraction. This behavior suggests that lutein molecules intercalate between carrageenan and inulin chains, disrupting their regular packing and hydrogen-bonding network. Importantly, no sharp peaks corresponding to crystalline lutein appeared in the H+L pattern, confirming that lutein was molecularly dispersed or present in an amorphous state within the hydrogel matrix—a desirable feature for controlled release applications.<br />FTIR Analysis: FTIR spectra revealed a broad band at 3200–3400 cm⁻¹ assigned to O–H stretching vibrations of hydrogen-bonded hydroxyl groups. The intensity of this band was higher in the H sample compared to pure carrageenan or inulin, indicating strong inter-polysaccharide hydrogen bond formation. In the H+L sample, the intensity decreased, suggesting partial disruption of the hydrogen-bonded network due to lutein-polysaccharide hydrophobic interactions and/or steric effects. Characteristic bands for sulfate groups (S=O asymmetric stretching at 1220–1260 cm⁻¹), C–O–C stretching (1030 cm⁻¹), and carboxylate (–COO⁻) symmetric/asymmetric stretching (1410 and 1600 cm⁻¹) were retained but exhibited reduced intensities in H+L, implying that lutein interacts with polar groups of the polysaccharide matrix. The absence of new peaks further supports the molecular dispersion of lutein without crystalline phase separation.<br />SEM Morphology: SEM micrographs showed that the lutein-free hydrogel (H) possessed a compact, smooth, and relatively uniform structure with finely dispersed small pores, indicative of a stable and well-organized gel network. In contrast, the lutein-loaded hydrogel (H+L) displayed a significantly rougher, more heterogeneous, and highly porous microstructure with larger voids and thinner pore walls. This morphological transformation is attributed to the disorganizing effect of lutein on chain packing and the reduction of inter-chain interactions, leading to a more open and permeable network—a structure that can facilitate diffusion-controlled release of the encapsulated bioactive.<br />Storage Stability: In the control sample (lutein/Tween 80), approximately 50% of lutein degraded after 30 days, confirming the inability of conventional micelles to provide an effective barrier against oxygen and light. In contrast, all hydrogel formulations significantly retarded lutein degradation. After 30 days, residual lutein was 65% for the 15% inulin hydrogel, 72% for the 20% inulin hydrogel, and 78% for the 25% inulin hydrogel. The superior protection offered by the 25% inulin formulation is attributed to the highest network density, smallest pore size, lowest oxygen diffusivity, and formation of glassy or microcrystalline domains at high solid content, which collectively restrict molecular mobility and reduce oxidative attack. These results demonstrate that increasing inulin concentration directly and proportionally enhances the protective capacity of the hydrogel.<br /><br />Antioxidant Activity (DPPH Assay): Lutein exhibited a clear concentration-dependent DPPH radical scavenging activity. As lutein concentration increased from 10 to 50 mg/mL, the percentage inhibition rose from approximately 17% to 69%. The calculated IC50 value was 32.8 mg/mL, indicating moderate-to-strong antioxidant potency. This activity originates from the conjugated polyene backbone of lutein, which enables electron or hydrogen donation to stabilize DPPH radicals, and from the terminal hydroxyl groups that further stabilize the resulting lutein radical. The obtained IC50 falls within the range previously reported for natural lutein from various botanical sources.<br /><br />Conclusion:<br />This study successfully demonstrated that the ι-carrageenan-inulin hybrid hydrogel, particularly at 25% inulin concentration, serves as an effective encapsulation system for lutein, offering significantly enhanced encapsulation efficiency and long-term chemical stability compared to conventional surfactant-based delivery systems. Structural analyses (XRD, FTIR, SEM) confirmed that lutein was molecularly dispersed within the predominantly amorphous, porous, and hydrogen-bonded polysaccharide network without forming crystalline aggregates. Increasing inulin concentration from 15% to 25% systematically improved both EE% (from 71% to 82%) and residual lutein after 30-day storage (from 65% to 78%), owing to enhanced network compactness, reduced oxygen permeability, and restricted molecular mobility. Lutein retained its intrinsic concentration-dependent antioxidant activity (IC50 = 32.8 mg/mL) after encapsulation. These findings establish the ι-carrageenan-inulin (25% inulin) hydrogel as a scalable, biocompatible, and biodegradable carrier with strong potential for incorporation into functional food matrices (e.g., fortified beverages, dairy desserts), pharmaceutical ophthalmic formulations, and cosmetic anti-aging/sunscreen products. Future studies should investigate the rheological behavior, digestibility, and in vivo bioavailability of lutein-loaded hydrogels under simulated gastrointestinal conditions, as well as the potential synergistic effects with other lipophilic bioactive compounds.هدف از پژوهش، طراحی هیدروژل هیبریدی یوتا-کاراگینان-اینولین به عنوان سامانه حامل برای لوتئین و بررسی تأثیر غلظت اینولین (15، 20 و 25 درصد وزنی/وزنی) بر پایداری ساختاری، راندمان درونپوشانی و فعالیت آنتیاکسیدانی بود. نتایج پراش پرتو ایکس (XRD) وجود یک هاله پهن در محدوده ۲θ=18-25° را نشان داد که حاکی از ساختار غالباً آمورف همراه با دامنههای نیمهبلورین محدود بود. افزودن لوتئین به طور معنیداری شدت هاله پراش را کاهش داد (05/0>p) که بیانگر کاهش کریستالینیتی و افزایش فاز آمورف است. طیفسنجی فروسرخ (FTIR) تشکیل پیوندهای هیدروژنی بین اجزاء را تأیید کرد و کاهش شدت باندهای –OH در نمونه حاوی لوتئین، نشاندهنده برهمکنش لوتئین با ماتریس هیدروژل بود. تصاویر میکروسکوپ الکترونی روبشی (SEM) نشان داد که نمونه هیدروژل فاقد لوتئین دارای ساختاری فشرده و یکنواخت بوده و نمونه حاوی لوتئین به طور قابل توجهی متخلخلتر و ناهمگنتر بود. راندمان درونپوشانی لوتئین در هیدروژلهای حاوی 25% اینولین به حدود ۸۲ درصد رسید که به طور معنیداری بیشتر از نمونههای 15% (۷۱ درصد) و 20% (۷۶ درصد) بود (05/0>p). بررسی پایداری طی ۳۰ روز نگهداری نشان داد که هیدروژل حاوی 25% اینولین بهترین عملکرد حفاظتی را داشته و حدود ۷۸ درصد لوتئین اولیه را حفظ کرد، در حالی که این مقدار در نمونه کنترل (لوتئین در توئین ۸۰) تنها ۵۰ درصد بود. فعالیت آنتیاکسیدانی لوتئین با آزمون DPPH نمایانگر قدرت آنتیاکسیدانی متوسط تا نسبتاً قوی بود. به طور کلی، هیدروژل یوتا-کاراگینان-اینولین با غلظت 25% اینولین به عنوان یک سامانه مؤثر برای درونپوشانی، حفاظت و انتقال لوتئین در کاربردهای غذایی، دارویی پیشنهاد میشود.https://foodresearch.tabrizu.ac.ir/article_21817_ef292ab6b47c336b2b31e7aba9db1ad4.pdf