Search Results (47)

Objectives: This study aimed to develop and validate a sensitive and reliable liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the quantification of flualprazolam and isotonitazene in serum samples to address critical gaps in forensic and clinical toxicology. Materials and Methods: A single-center validation study was conducted using serum samples spiked with analyte standards. Analytical parameters, including linearity, precision, recovery, the limit of detection (LOD), and the limit of quantification (LOQ), were evaluated in accordance with international guidelines. The LC-MS/MS method employed a Shimadzu Triple Quadrupole™ MS 8045 system with solid-phase extraction (SPE) for sample preparation. Results: The method exhibited high linearity for flualprazolam (r² = 0.997) and isotonitazene (r² = 0.999) over a concentration range of 1–100 ng/mL. The LODs were determined as 0.608 ng/mL and 0.192 ng/mL, and the LOQs were 1.842 ng/mL and 0.584 ng/mL for flualprazolam and isotonitazene, respectively. Recovery tests yielded results within the acceptable range of 70–120%. Flualprazolam demonstrated recovery rates of 98.0% and 97.0% at theoretical concentrations of 10 ng/mL and 50 ng/mL, respectively. In contrast, the isotonitazene recovery rates were slightly lower, measuring 75.5% at 10 ng/mL and 71.9% at 50 ng/mL, suggesting minor matrix effects that could influence its quantification. Precision analysis, including both repeatability and reproducibility, highlighted the reliability of the method. The %RSD values for flualprazolam were consistently below 7.07%, with mean concentrations closely aligning with theoretical values across fortification levels. For isotonitazene, the %RSD values remained below 6.24%, although recoveries at higher concentrations indicated potential challenges in matrix interaction. Conclusions: This validated LC-MS/MS method offers high sensitivity, precision, and recovery for detecting flualprazolam and isotonitazene in serum, filling a critical need in toxicological investigations. Further validation in other biological matrices is recommended to broaden its applicability. Full article

Potassium iodate and potassium iodide are commonly fortified in iodized table salt, which must be continuously monitored to maintain quality. Our study reported an optimized detection method for total iodine in iodized table salt using 0.5 M sodium bisulfite as the reducing agent. The iodized table salt (0.5 g) was dissolved in 0.5 M sodium bisulfite solution prior to injection in ultra-high-performance liquid chromatography (UHPLC) coupled with a diode array detector using a weak anion-exchange column (2.1 mm × 150 mm, 5 μm). Iodide was eluted at 9.92 ± 0.06 min (λ = 223 nm) when an isocratic mobile phase of 1:1 (v/v) methanol/120 mM phosphate buffer mixed with tetrasodium pyrophosphate (pH 3.0) was running at 0.20 mL/min (15 min). Iodide was detected as total iodine from 10.0 to 50.0 mg/kg with a limit of detection (LOD) of 1.2 mg/kg and a limit of quantification (LOQ) of 3.7 mg/kg. The method was validated with relative standard deviations (RSDs) of 4.2%, 0.4%, 1.6%, and 0.8% for accuracy, repeatability, intermediate precision, and robustness, respectively. The determination of total iodine was successful on six (6) samples (n = 3), which recovered 87.2–106.9% of iodate and iodide spike. Thus, this study provides a validated protocol for the determination of total iodine in iodized table salt using 0.5 M sodium bisulfite. Full article

Background/Objectives: The dietary changes experienced by residents in long-term care facilities (LTCFs) following an earthquake are poorly understood. This study aimed to examine variations in nutritional status among residents of an Integrated Facility for Medical and Long-term Care (IFMLC), a particular type of Japanese LTCF, after the 2024 Noto Peninsula Earthquake. Methods: This descriptive study was conducted at the single IFMLC. A total of 115 residents living at the facility on 1 January 2024, at the time of the earthquake, were recruited for the study. The focus was the body weight and skeletal muscle mass changes observed before and after the earthquake. The observation period lasted for three months following the earthquake. Results: Water outage persisted for over a month, making dishwashing impossible and leading to an extended reliance on disposable dishes with limited capacity. This situation consequently reduced the variety and volume of meal options and overall energy intake meals. Residents’ body weight significantly decreased 3 months after the earthquake, and the prevalence of weight loss and skeletal muscle mass loss was particularly high in residents with normal swallowing function. To address nutritional deficiencies post-earthquake, the registered dietitian enhanced energy sufficiency through food fortification, oral nutritional supplements, and pre-prepared ready-to-hang liquid formulas. Conclusions: To prevent further weight and skeletal muscle mass reduction among IFMLC residents, providing ample water, and a disaster manual that can be used even with limited resources is essential. Furthermore, preparing for disasters by stockpiling foods and implementing strategies to enhance energy sufficiency is crucial. Full article

In recent decades, there has been growing interest in the fortification of food products with antioxidants and phenolics derived from plant by-products. The present study focused on the production of a plant-based beverage enriched with hazelnut cuticle extract to characterize its antioxidant content, phenolic profile, and organoleptic characteristics. Liquid chromatography-mass spectrometry (LC-MS) enabled the identification of key polyphenols in hazelnut cuticles, including catechin, epicatechin, and quercetin derivatives, guiding the selection of a biocompatible Natural Deep Eutectic Solvent (NADES) composed of choline chloride and lactic acid for efficient extraction. The obtained phytochemical profile of the extract revealed a high concentration of bioactive compounds, with a Total Phenolic Content of 160.88 ± 14.27 mg GAE/g and Antioxidant Power measured by DPPH of 5848.2 ± 11.3 μmol TE/g. The bioaccessibility of phenolics in the fortified hazelnut-based beverage was determined after in vitro digestion, reaching a value of 89.7%, indicating excellent release and stability during digestion. Organoleptic evaluation revealed high sensory acceptability, with aftertaste scoring 3.61 ± 0.4 respect the 3.94 ± 1.3 result of reference milk, on a 5-point scale. In conclusion, this study demonstrates the potential for sustainable valorization of hazelnut cuticles, through their incorporation as NADES extracts in plant-based milk, providing an innovative solution to reduce food waste while catering to consumer demand for nutritionally enriched and eco-friendly products. Full article

The in situ fortification of vitamin B12 (VB12) in foods through fermentation is an effective strategy to address the deficiency of this micronutrient, and precise monitoring of VB12 production is crucial for developing VB12-fortified functional foods. Liquid chromatography–tandem mass spectrometry (LC-MS/MS) is advantageous for analyzing trace substances in food due to its high sensitivity. In the present study, an LC-MS/MS method capable of rapidly and accurately quantifying three active forms of VB12, namely adenosylcobalamin (AdoCbl), methylcobalamin (MeCbl), hydroxocobalamin (OHCbl), in 8 min were developed. Meanwhile, the quantitative result of this method is not affected by pseudo-VB12 because the selected ion channels include fragments of active VB12. Maintaining light-shielding during extraction and purification is essential, as light exposure during the process can decrease the content of detected VB12 by about 30%. At last, the developed method was applied for the determination of VB12 in fermented rice bran and the cell mass of Propionibacterium freudenreichii. The results showed that AdoCbl was the predominant form of VB12 during fermentation, and the addition of cobalt did not influence the proportions of the three VB12 types. The present study reported a rapid and accurate method for the simultaneous determination of three active forms of VB12, which can effectively support the development of foods with VB12 fortification. Full article

This study aimed to enhance the nutritional value of jellies by fortification with polyphenol extracts derived from Pinus sylvestris L. shoots at various maturation stages. Pinus sylvestris L., a coniferous species, is widely used in traditional medicine and functional foods due to its antioxidant, anti-inflammatory, and antimicrobial properties. Its needles, bark, and shoots are commonly used to extract bioactive compounds such as phenolic acids and flavonoids. In the current study, extracts were derived from young shoots collected directly from natural forest environments and processed using a decoction method to preserve bioactive compounds. The novel jelly formulations were prepared using pine shoots harvested at three maturity stages: stage I (4 cm), stage II (8 cm), and stage III (12 cm). All determinations were conducted both on the pure decoction extracts and the jelly samples to ensure a comprehensive analysis. High-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS) allowed the identification of eight phenolic acids and six flavonoids in the samples. Significant differences were observed between the pine shoot extracts and jellies at different development stages. Notably, stage II exhibited optimal polyphenol content (312.2 mg GAE/100 g), DPPH free radical scavenging activity (94.9%), dry matter content (79.5%), and acidity (0.79% citric acid/g). A similar pattern emerged in the jelly samples (jelly2 (pine decoction stage II) > jelly1 (pine decoction stage I) > jelly3 (pine decoction stage III)). All extracts demonstrated antioxidant potential in DPPH free radical quenching assays. FTIR analysis evaluated structural changes in phenolic compounds during jelly formulation, focusing on key absorption bands at 1600 cm⁻¹ (C=C stretching) and 3336 cm⁻¹ (-OH stretching) using a Shimadzu IR Prestige-21 spectrophotometer. Compared to extracts, jellies showed diminished band intensities, indicating thermal degradation of phenolic compounds during processing. This aligns with observed reductions in antioxidant capacity and phenolic content, suggesting partial destabilization of these bioactive compounds. However, their integration into the jelly matrix highlights the potential for functional applications. The textural attributes of jellies were also assessed, and differences were attributed to the changes in acidity and moisture content of the pine shoots during maturation. Pine shoot extracts at specific maturation stages are valuable sources of antioxidant and polyphenol compounds and were successfully employed in functional applications belonging to the food or nutraceutical industry. Full article

Background/Objectives: The widespread use of antibiotics, which wastewater treatment plants (WWTPs) cannot fully remove, in human and veterinary medicine leads to their release into wastewater, resulting in the contamination of aquatic environments. Bivalves can accumulate these antibiotics, posing a risk to shellfish consumers, including potential antimicrobial resistance. This study aimed to assess how three cooking methods—marinating, steaming, and grilling—affect the concentration of 33 different antibiotics in bivalves fortified at the level of maximum residue limit (MRL) and twice the MRL (2MRL). Results: The data show the percentage of antibiotic remaining after cooking: 100% indicates stability or no reduction; values above 100% show an increase in concentration, and values below 100% reflect a decrease in antibiotic concentration. In general, all culinary procedures removed part of the added antibiotics. However, the most effective method was marinating (47%), followed by steaming (60%) and finally grilling (92%). It was also found that, overall, the fortification level, MRL or 2MRL, did not impact antibiotic removal in each cooking method. Moreover, different antibiotics’ classes presented diverse removals when cooked, ranging between 0% for penicillins and 73% for sulphonamides. Furthermore, the results showed a great diversity of responses to cooking within some antibiotic classes. Methods: After cooking, the analysis was based on solid–liquid extraction followed by liquid chromatography–quadrupole time-of-flight mass spectrometry (UHPLC-ToF-MS). Conclusions: The ongoing monitoring of antibiotic levels is essential, and further research is needed to understand how cooking affects these substances and their metabolites. This will help assess the real risk to consumers and guide risk-mitigation measures. Full article

Along the food production chain of animal, fish, and vegetable products, a huge amount of by-products are generated every year. Major nutritional, financial, and environmental advantages can be achieved by transforming them into functional ingredients for food formulation and fortification. In this review, we investigated various conventional and emerging treatments recently employed to obtain functional ingredients rich in proteins, fibers, and bioactive compounds from vegetables, fish, meat, and dairy by-products. The optimal enrichment level in food as well as the nutritional, techno-functional, and sensory properties of the final food were also discussed. Novel technologies such as ultrasounds, microwaves, and high pressure have been successfully adopted to enhance the extraction of target compounds. The functional ingredients, added both in liquid or powder form, were able to improve the nutritional quality and antioxidant potential of food, although high levels of fortification may cause undesired changes in texture and flavor. This review provides important considerations for further industrial scale-up. Full article

Vitamin D₃ deficiency is a global phenomenon, which can be managed with supplementation and food fortification. However, vitamin D₃ bioaccessibility may depend on factors such as matrix composition and interactions throughout the gastrointestinal (GI) tract. This research focused on the effect of different matrices on vitamin D₃ content during digestion, as well as the effect of pH on its bioaccessibility. The INFOGEST protocol was employed to simulate digestion. Three different types of commercial supplements, two foods naturally rich in vitamin D₃, and three fortified foods were investigated. High-Performance Liquid Chromatography was used to determine the initial vitamin D₃ content in the supplements and foods, as well as after each digestion stage. The results indicate that the foods exhibited higher bioaccessibility indices compared to the supplements and a higher percentage retention at the end of the gastric phase. The pH study revealed a positive correlation between an increased gastric pH and the corresponding content of vitamin D₃. Interestingly, exposing the matrix to a low pH during the gastric phase resulted in an increased intestinal content of D₃. Vitamin D₃ is more bioaccessible from foods than supplements, and its bioaccessibility is susceptible to changes in gastric pH. Fasting conditions (i.e., gastric pH = 1) enhance the vitamin’s bioaccessibility. Full article

The objective of this study was to determine the optimal conditions for the recovery of bioactive and antioxidant compounds in aqueous solutions of Cistus creticus leaves and then employ the optimal extract for the enrichment of yogurt samples. The optimal conditions were established by a response surface methodology and were determined to be a liquid-to-solid ratio of 48 mL/g at 76 °C for 41 min. The optimum extract yielded TPC 157.17 mg GAE/g dw and TFC 2.38 mg QE/g dw, while FRAP and DPPH values were 1258.52 and 933.67 μmol AAE/g dw, respectively. HPLC-DAD was utilized to identify and quantify specific polyphenols, like myricetin rhamnoside, in the extract. The optimal extract was then added to yogurt desserts during their preparation at three different concentrations to study how the physicochemical characteristics of the yogurt, as well as the antioxidant capacity added during enrichment, were affected. Statistical analysis of the results was carried out in order to obtain more valid data. It seems that the most suitable concentration for yogurt fortification was 0.1% w/v of the extract as, at this concentration, the yogurts exhibited higher antioxidant capacity, and their physicochemical characteristics were improved. Full article

Marigold (Tagetes spp.) flower petals are the most vital sources of carotenoids, especially lutein esters, for the production of natural lutein to use for food, feed, and pharmaceutical industries. Several marigold cultivars are cultivated globally; however, their lutein ester composition and contents have not been widely investigated. Considering this, this study aimed to identify and quantify prominent carotenoid esters from the flower petals of ten marigold cultivars by liquid chromatography (LC)–diode-array detection (DAD)–mass spectrometry (MS). In addition, tocopherols, phytosterols, and fatty acids were analyzed by gas chromatography (GC)–flame ionization detection (FID) and GC–MS. Furthermore, the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS^•+) and 2,2-diphenyl-1-picrylhydrazyl (DPPH^•) radical scavenging abilities of lipophilic extracts were determined. The total carotenoid contents varied significantly (p < 0. 05, Tukey HSD) among cultivars, ranging from 25.62 (cv. Alaska)–2723.11 µg/g fresh weight (cv. Superboy Orange). Among the five major lutein-diesters, (all-E)-lutein-3-O-myristate-3′-O-palmitate and lutein dipalmitate were predominant. Among the studied cultivars, α-tocopherol was recorded, ranging from 167.91 (cv. Superboy Yellow) to 338.50 µg/g FW (cv. Taishan Orange). Among phytosterols, β-sitosterol was the most prevalent phytosterol, ranging between 127.08 (cv. Superboy Yellow) and 191.99 µg/g FW (cv. Taishan Yellow). Palmitic acid (C_16:0; 33.36–47.43%) was the most dominant among the fatty acids. In this study, the highest contents of lutein were recorded from cv. Superboy Orange; however, due to the substantially higher flower petal yield, the cv. Durango Red can produce the highest lutein yield of 94.45 kg/ha. These observations suggest that cv. Durango Red and cv. Superboy Orange are the ideal candidates for lutein fortification in foods and also for commercial lutein extraction. Full article

The aim of this research was to evaluate the levels of antioxidants and polyphenols in pasta enriched with either carrot or olive leaf flours after simulating gastrointestinal digestion. Pasta samples were prepared with fixed amounts of carrot and olive leaf flours (15% and 6% of the total mixture, respectively). We measured the antioxidant capacity and polyphenol content at different stages of the pasta production process, starting from the initial flour to the cooked pasta, and tested samples of the liquid component and solid waste resulting from the digestion process. The antioxidant activity was measured by the FRAP method, while the polyphenol content was measured by the Folin–Ciocalteu method. Vitamin E contents were measured by HPLC. The pasta enriched with carrot (1.26 ± 0.05 mmol/100 g) and olive leaf (2.9 ± 0.07 mmol/100 g) exhibited higher antioxidant power compared to the unenriched pasta (0.8 ± 0.1 mmol/100 g). The polyphenol content followed a similar trend, with values of 131.23 ± 3.08 for olive flour-enriched pasta, 79.15 ± 1.11 for carrot flour-enriched pasta, and 67.5 ± 1.39 for the wheat-only pasta. The pasta samples maintained their antioxidant and polyphenol levels even after undergoing the simulated digestion process. Significantly, the liquid component of the pasta with olive leaf flours had the highest levels of antioxidants and polyphenols during all stages of the digestion process. According to the results of this study, pasta enriched with carrot and olive leaf flours shows promising potential for improving nutritional and functional properties by increasing antioxidant and polyphenol content. The samples were also evaluated by a sensory panel, which showed that fortification modified the perception of some organoleptic attributes without affecting the overall taste of the pasta. Full article

Organic heme iron in the form of heme iron polypeptide (HIP) is a bioavailable form of iron that can be used for dietary supplements. However, one practical challenge with HIP is that the quality of HIP prepared with different batches of raw material could lead to HIP products with inconsistent characteristics. In this study, skipjack tuna blood, a by-product in canned tuna industry, was converted to HIP at different degrees of enzymatic hydrolysis. The variation in HIP physical–chemical characteristics from different batches was evaluated, including composition, solubility, and molecular weight distribution. It was found that the batch variation had no effect on HIP composition and solubility; however, the degree of hydrolysis (DH) and the size of peptides that interact with heme greatly influenced HIP solubility at pH 2. Tuna-HIP with a low DH (DH, 8%) had 1.76-fold greater solubility than tuna-HIP with a high DH (DH, 32%). High-performance liquid chromatography (HPLC) revealed that tuna-HIP with a low DH had a molecular weight ranging from 1 kDa to 5 kDa. In summary, HIP-derived tuna blood was found to contain 70.54 ± 3.22 mg/100 g of iron and exhibit good solubility at 58.0 ± 2.16% at pH 2. Thus, tuna-HIP with a low DH might be a suitable functional ingredient for iron fortification of food. Full article

The incorporation of fermented camel milk with natural additives possesses numerous benefits for the treatment of various pathological and metabolic conditions. The present study investigated the impact of fortification of fermented camel milk with sage or mint leaves powder (1 and 1.5%, respectively) on glucose and insulin levels, lipid profile, and liver and kidney functions in alloxan-induced diabetic rats. The gross chemical composition of sage and peppermint leaves powder was studied. The chemical composition of sage and mint extracts was performed using liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-MS) of sage and mint extracts. Furthermore, a total of forty-two adult normal male albino rats were included in this study, whereas one group was kept as the healthy control group (n = 6 rats) and diabetes was induced in the remaining animals (n = 36 rats) using alloxan injection (150 mg/kg of body weight). Among diabetic rats groups, a control group (n = 6 rats) was kept as the diabetic control group whereas the other 5 groups (6 rats per group) of diabetic rats were fed fermented camel milk (FCM) or fermented camel milk fortified with 1 and 1.5% of sage or mint leaves powder. Interestingly, the oral administration of fermented camel milk fortified with sage or mint leaves powder, at both concentrations, caused a significant decrease in blood glucose level and lipid profile, and an increase in insulin level compared to the diabetic control and FCM groups. Among others, the best results were observed in the group of animals that received fermented camel milk fortified with 1.5% sage powder. In addition, the results revealed that the fermented camel milk fortified with sage or mint leaves powder improved the liver and kidney functions of diabetic rats. Our study concluded that the use of sage and mint leaves powder (at a ratio of 1.5%) with fermented camel milk produces functional food products with anti-diabetic activity. Full article

In this study, naked oat bran albumin hydrolysates (NOBAH) were subjected to gel chromatography with Sephadex G-15, reverse phase-high liquid performance separation, and UPLC-ESI-MS/MS identification. Six safe peptides including Gly-Thr-Thr-Gly-Gly-Met-Gly-Thr (GTTGGMGT), Gln-Tyr-Val-Pro-Phe (QYVPF), Gly-Ala-Ala-Ala-Ala-Leu-Val (GAAAALV), Gly-Tyr-His-Gly-His (GYHGH), Gly-Leu-Arg-Ala-Ala-Ala-Ala-Ala-Ala-Glu-Gly-Gly (GLRAAAAAAEGG), and Pro-Ser-Ser-Pro-Pro-Ser (PSSPPS) were identified. Next, in silico screening demonstrated that QYVPF and GYHGH had both angiotensin-I-converting enzyme (ACE) inhibition activity (IC₅₀: 243.36 and 321.94 μmol/L, respectively) and Zinc-chelating ability (14.85 and 0.32 mg/g, respectively). The inhibition kinetics demonstrated that QYVPF and GYHGH were both uncompetitive inhibitors of ACE. Molecular docking showed that QYVPF and GYHGH could bind, respectively, three and five active residues of ACE with short hydrogen bonds (but not belonging to any central pocket). QYVPF and GYHGH could bind, respectively, twenty-two and eleven residues through hydrophobic interactions. Moreover, GYHGH was able to affect zinc tetrahedral coordination in ACE by interacting with His383. The inhibition activities of QYVPF and GYHGH toward ACE were relatively resistant to gastrointestinal digestion. GYHGH improved zinc solubility in the intestines (p > 0.05) because its amino and carboxyl groups were chelating sites for zinc ions. These results suggest the potential applications of naked oat peptides for potential antihypertension or zinc fortification. Full article