Using a DNA mini-barcode within the ITS region to identify toxic Amanita in mushroom poisoning cases.

Mushroom poisoning contributes significantly to global foodborne diseases and related fatalities. Amanita mushrooms frequently cause such poisonings; however, identifying these toxic species is challenging due to the unavailability of fresh and intact samples. It is often necessary to analyze residues, vomitus, or stomach extracts to obtain DNA sequences for the identification of species responsible for causing food poisoning. This usually proves challenging to obtain usable DNA sequences that can be analyzed using conventional molecular biology techniques. Therefore, this study aimed to develop a DNA mini-barcoding method for the identification of Amanita species. Following the evaluation and optimization of universal primers for DNA mini-barcoding in Amanita mushrooms, we found that the internal transcribed spacer (ITS) gene sequence primer ITS-a was the most suitable DNA barcode primer for identifying Amanita species. Forty-three Amanita samples were subsequently amplified and sequenced. The sequences obtained were analyzed for intra- and inter-species genetic distances, and a phylogenetic tree was constructed. The findings indicated that the designed primers had strong universality among the Amanita samples and could accurately identify the target gene fragment with a length of 290 bp. Notably, the DNA mini-barcode accurately identified the 43 Amanita samples, demonstrating high consistency with the conventional DNA barcode. Furthermore, it effectively identified DNA from digested samples. In summary, this DNA mini-barcode is a promising tool for detecting accidental ingestion of toxic Amanita mushrooms. It may be used as an optimal barcode for species identification and traceability in events of Amanita-induced mushroom poisoning. KEY POINTS: • Development of a DNA mini-barcoding method for Amanita species identification without fresh samples. • The ITS-a primer set was optimized for robust universality in Amanita samples. • The mini-barcode is suitable for screening toxic mushroom species in mushroom poisoning cases.

Development of a sandwich enzyme-linked immunosorbent kit for reliable detection of milk allergens in processed food.

The inclusion of undeclared cow's milk proteins may cause health complications to milk-allergic consumers and is one of the leading cause of food recall in many countries all over the world. Therefore, to keep control on such incidences in processed products, we established a milk sandwich ELISA test kit by incorporating two polyclonal antibodies against milk proteins obtained from different species. Its analytical effectiveness in terms of sensitivity, specificity, accuracy, trueness, and precision were all analyzed. The limit of detection (LOD) of the test kit was 0.011 ppm, with high specificity for milk protein residues. The test kit was highly specific, apart from considerable cross-reactivity with goat milk and minor cross-reactivity with donkey and horse milk. The coefficient of variation of the test kit for intra-assay ranged from 4.02% to 14.62% and inter-assay ranged from 6.05% to 15.08% respectively. The sandwich ELISA was highly specific in detecting commercial food products. In a limited retail survey, 5/6 of the milk proteins declared on the ingredient labels tested positive for milk proteins. The study offers effective technical support for the sensitive detection of milk products both for food manufacturers and regulatory authorities.

Development of a sensitive sandwich-ELISA assay for reliable detection of fish residues in foods.

A new sandwich-type Enzyme-Linked Immunosorbent Assay (ELISA) method was developed based on goat IgG as capturing antibody and rabbit IgG as detecting antibody targeting soluble antigenic fish proteins in foods as detection targets. The assay has provided a relatively lower limit of quantitation (LoQ) for fish proteins with LoQ 0.5 ng/ml and appears highly sensitive. The analysis of 24 different substances, both raw and boiled, revealed no cross-reactivity above the cut-off point of the limit of quantitation. Recoveries of the SB spiked food matrixes were in the range of 83-131%. Assay precision testing proved that repeatability (<5%) and reproducibility (<11%) had an acceptable level of variation. The sandwich ELISA was capable of detecting all tested commercially important fish. As a potential analytical tool, the newly developed immunoenzymatic method is suitable for detecting undeclared fish residues in real food samples available in the market, thereby will help to reduce the incidents of fish allergies.

Rapid Classification and Quantification of Camellia (Camellia oleifera Abel.) Oil Blended with Rapeseed Oil Using FTIR-ATR Spectroscopy.

Currently, the authentication of camellia oil (CAO) has become very important due to the possible adulteration of CAO with cheaper vegetable oils such as rapeseed oil (RSO). Therefore, we report a Fourier transform infrared (FTIR) spectroscopic method for detecting the authenticity of CAO and quantifying the blended levels of RSO. In this study, two characteristic spectral bands (1119 cm-1 and 1096 cm-1) were selected and used for monitoring the purity of CAO. In combination with principal component analysis (PCA), linear discriminant analysis (LDA), and partial least squares regression (PLSR) analysis, qualitative and quantitative methods for the detection of camellia oil adulteration were proposed. The results showed that the calculated I1119/I1096 intensity ratio facilitated an initial check for pure CAO and six other edible oils. PCA was used on the optimized spectral region of 1800-650 cm-1. We observed the classification of CAO and RSO as well as discrimination of CAO with RSO adulterants. LDA was utilized to classify CAO from RSO. We could differentiate and classify RSO adulterants up to 1% v/v. In the quantitative PLSR models, the plots of actual values versus predicted values exhibited high linearity. Root mean square error of calibration (RMSEC) and root mean square error of cross validation (RMSECV) values of the PLSR models were 1.4518%-3.3164% v/v and 1.7196%-3.8136% v/v, respectively. This method was successfully applied in the classification and quantification of CAO adulteration with RSO.

Diversity in storage age enables discrepancy in quality attributes and metabolic profile of Citrus grandis "Tomentosa" in China.

The folk proverb "the older, the better" is usually used to describe the quality of Citrus grandis "Tomentosa" (CGT) in China. In this study, CGT aged for 6-, 12-, 16-, and 19-years were collected for the investigation of infusion color, main bioactive components, antioxidant activity, metabolic composition, and pathway. The results found that infusion color, the total phenolic and flavonoid, and antioxidant activity of CGT were obviously changed by aging process. Through untargeted metabolomics, 55 critical metabolites were identified to in discrimination of CGT with different storage ages, mainly including phenylpropanoids, lipids, and organic oxygen compounds. Twenty compounds that showed good linear relationships with storage ages could be used for year prediction of CGT. Kyoto encyclopedia of genes and genomes enrichment pathway analysis uncovered important metabolic pathways related to the accumulation of naringin, kaempferol, and choline as well as the degradation of benzenoids, thus supporting that aged CGT might be more beneficial to health. Correlation analysis provided that some key metabolites with bitter taste and biological activity were involved in the darkening and reddening of CGT infusion during aging, and total phenolic and flavonoid were more strongly associated with the antioxidant activity of CGT. This study systematically revealed the quality changes and key metabolic pathways during CGT aging at first time. PRACTICAL APPLICATION: This study reveals the differences in quality attributes and metabolic profile between CGT with different storage ages, providing guidance for consumers' consumption, and also providing more scientific basis for the quality evaluation and improvement of CGT.

Screening of specific binding peptide for ß-lactoglobulin using phage display technology.

ß-lactoglobulin (ß-Lg) is a major food allergen, there is an urgent need to develop a rapid method for detecting ß-Lg in order to avoid contact or ingestion by allergic patients. Peptide aptamers have high affinity, specificity, and stability, and have broad prospects in the field of rapid detection. Using ß-Lg as the target, this study screened 11 peptides (P1-11) from a phage display library. Using molecular docking technology to predict binding energy and binding mode of proteins and peptides. Select the peptides with the best binding ability to ß-Lg (P5, P7, P8) through ELISA. Combining them with whey protein, casein, and bovine serum protein, it was found that P7 has the best specificity for ß-Lg, with an inhibition rate of 87.99%. Verified by molecular dynamics that P7 binds well with ß-Lg. Therefore, this peptide can be used for the recognition of ß-Lg, becoming a new recognition element for detecting ß-Lg.

Revealing oxidative degradation of lipids and screening potential markers of four vegetable oils during thermal processing by pseudotargeted oxidative lipidomics.

The oxidative degradation of lipids in vegetable oils during thermal processing may present a risk to human health. However, not much is known about the evolution of lipids and their non-volatile derivatives in vegetable oils under different thermal processing conditions. In the present study, a pseudotargeted oxidative lipidomics approach was developed and the evolution of lipids and their non-volatile derivatives in palm oil, rapeseed oil, soybean oil, and flaxseed oil under different thermal processing conditions was investigated. The results showed that thermal processing resulted in the oxidative degradation of TGs in vegetable oils, which generated oxTGs, DGs, and FFAs, as well as TGs with smaller molecular weights. The lower the fatty acid saturation, the more severe the oxidative degradation of vegetable oils and thermal processing at high temperatures should be avoided if possible. From the accumulation of oxTGs concentrations, the hazards during thermal processing at high temperatures were, in descending order, soybean oil, rapeseed oil, flaxseed oil, and palm oil. The non-volatile potential markers were screened in palm oil, rapeseed oil, soybean oil, and flaxseed oil for 1, 7, 5, and 2 markers related to thermal processing time, respectively. The study provided suggestions for the consumption of vegetable oils from multiple perspectives and identified markers for monitored oxidative degradation of vegetable oils.

An instrument-free, integrated micro-platform for rapid and multiplexed detection of dairy adulteration in resource-limited environments.

In dairy industry, expensive yak's milk, camel's milk, and other specialty dairy products are often adulterated with low-cost cow's milk, goat's milk and so on. Currently, the detection of specialty dairy products typically requires laboratory settings and relies on skilled operators. Therefore, there is an urgent need to develop a multi-detection technology and on-site rapid detection technique to enhance the efficiency and accuracy of the detection of specialty dairy products. In this study, we introduced a fully integrated and portable microfluidic detection platform called Sector Self-Driving Microfluidics (SDM), designed to simultaneously detect eight common species-specific components in milk. SDM integrated nucleic acid extraction, purification, loop-mediated isothermal amplification (LAMP), and lateral flow strip (LFS) detection functions into a closed microfluidic system, enabling contamination-free visual detection. The SDM platform used a constant-temperature heating plate, powered by a mobile battery, eliminated the need for additional power support. The SDM platform achieved nucleic acid enrichment and transfer through magnetic force and liquid flow driven by capillary forces, operating without external pumps. The standalone SDM platform could detect dairy components with as low as 1% content within 1 h. Validation with 35 commercially available samples demonstrated 100% specificity and accuracy compared to the gold standard real-time PCR. The SDM platform provided the dairy industry with an efficient, convenient, and accurate detection tool, enabling rapid on-site testing at production facilities or sales points. This facilitated real-time monitoring of quality issues during the production process, quickly identifying potential risks and preventing substandard products from entering the market.

Characterization, purification of Poncirin from edible citrus Ougan (Citrus reticulate cv. Suavissima) and its growth inhibitory effect on human gastric cancer cells SGC-7901.

Poncirin is a bitter flavanone glycoside with various biological activities. Poncirin was isolated from four different tissues (flavedo, albedo, segment membrane, and juice sac) of Ougan fruit (Citrus reticulate cv. Suavissima). The highest content of poncirin was found in the albedo of Ougan fruit (1.37 mg/g DW). High speed counter-current chromatography (HSCCC) combined with D101 resin chromatography was utilized for the separation and purification of poncirin from the albedo of Ougan fruit. After this two-step purification, poncirin purity increased from 0.14% to 96.56%. The chemical structure of the purified poncirin was identified by both HPLC-PDA and LC-MS. Poncirin showed a significant in vitro inhibitory effect on the growth of the human gastric cancer cells, SGC-7901, in a dose-dependent manner. Thus, poncirin from Ougan fruit, may be beneficial for gastric cancer prevention. The purification method demonstrated here will be useful for further studies on the pharmacological mechanism of poncirin activity, as well as for guiding the consumption of Ougan fruit.

Oxidative lipidomics to elucidate the non-volatile derivatives of four typical triglycerides in vegetable oils under simulated frying conditions.

Vegetable oils with different saturations have varied composition of triglycerides (TGs) and produce different non-volatile derivatives during oxidation. Precise characterization of the non-volatile derivatives of TGs is essential for understanding the degradation of TGs and the production pattern of non-volatile derivatives. Oxidative lipidomics was combined with collision-induced dissociation and electron-activated dissociation to elucidate the precise structures of non-volatile derivatives produced under simulated frying conditions by 1,3-dipalmitoyl-2-oleoylglycerol (POP), triolein (OOO), trilinolein (LLL), and trilinolenin (LnLnLn). The results indicate that the unsaturated fatty acyl chains at the sn-2 position were more susceptible to oxidation compared with those at the sn-1/3 position. Species of non-volatile derivatives included epoxy-, hydroperoxy-, hydroxy-, and oxo-TGs, as well as degradation products. The potential reaction pathways of TGs and their non-volatile derivatives were also proposed. This study elucidated oxidative degradation mechanisms of the four typical TGs and provided a theoretical basis for changes of vegetable oils during frying.

New insight into the evolution of volatile profiles in four vegetable oils with different saturations during thermal processing by integrated volatolomics and lipidomics analysis.

To compare the oxidation products of four types of vegetable oils (palm oil, soybean oil rapeseed oil, and flaxseed oil) during thermal processing, lipidomics, volatolomics and simulation analyses were integrated to investigate the evolution of volatile profiles. The evolution of volatile profiles in different vegetable oils were found to be different, which are attribute to their different lipid composition. There were potential markers of palmitic acid-based vegetable oils as undecanal, dodecanal and 2-hexanone. A potential marker of oleic acid-based vegetable oils was 2-undecenal. (E,E)-2,4-nonadienal, 3-octen-2-one, and 3-nonen-2-one were potential markers of linoleic acid-based vegetable oils. The potential markers of linolenic acid-type vegetable oils were 1-penten-3-ol, (E)-2-butenal, (E)-2-pentenal, (E,E)-2,4-heptadienal (1), (E,E)-2,4-heptadienal (2), 2-ethyl-furan (1), 2-pentanone, and 3-hexen-2-one. The present study provides a new and comprehensive strategy to elucidate the changes of volatile compounds in thermal processed vegetable oil.

Effect of ultrahigh-pressure treatment on the structure and allergenicity of peach allergenic proteins.

Peach is a common plant-derived allergenic food and ultrahigh-pressure treatment is often used in peach products. In our study, an in-depth analysis of the structural and allergenicity changes of peach allergenic proteins after UHP treatment was performed by spectroscopy, mass spectrometry combined with serology and cytology. The results indicated that UHP treatment could reduce the content of peach soluble proteins and cause changes in secondary and tertiary structures. In addition, more hydrophobic residues were exposed and proteins tended to polymerize after UHP-treatment. The results of immunological assays showed that UHP treatment could reduce the IgE binding capacity of peach proteins and affect the ability of basophil degranulation, the upregulation of some cytokines may contribute to the reduction of peach protein allergenicity. Notably, UHP treatment may lead to the masking of some digestion sites in Pru p 3 epitopes, thus impeding human digestion and increasing the potential risk of allergenicity.

Integration of metabolomics and transcriptomics analyses reveals the mechanism of nano-selenium treated to activate phenylpropanoid metabolism and enhance the antioxidant activity of peach.

Foliar spraying to improve the quality of fruits is a general approach nowadays. In this study, 10 ppm nano-selenium (nano-Se) diluted with distilled water was sprayed on peach leaves every 10 days for a total of 7 sprays during the fruit set period. And then their fruit quality was compared with that of control group. It was found that the firmness, soluble solid concentration, total phenol, and proanthocyanidin content of the peaches were raised after the nano-Se treatment. Moreover, the ascorbic acid glutathione loop (ASA-GSH loop) was fully activated in the nano-Se treated group, and the associated antioxidant capacity and enzyme activity were significantly increased. Metabolomics revealed that nano-Se could upregulate some metabolites, such as phenylalanine, naringenin, and pinocembrin, to fully activate the metabolism of phenylpropanoids. Further, based on transcriptomics, nano-Se treatment was found to affect fruit quality by regulating genes related to phenylpropanoid metabolism, such as arogenate/prephenate dehydratase (ADT), genes related to abscisic acid metabolism such as (+)-abscisic acid 8'-hydroxylase (CYP707A), and some transcription factors such as MYB. Based on the comprehensive analysis of physicochemical indicators, metabolomics, and transcriptomics, it was found that nano-Se improved fruit quality by activating phenylpropanoid metabolism and enhancing antioxidant capacity. This work provides insights into the mechanism of the effect of nano-Se fertilizer on peach fruit quality. PRACTICAL APPLICATION: The firmness and soluble solid concentration of peaches are higher after nano-Se treatment, which is more in line with people's demand for hard soluble peaches like "Yingzui." The antioxidant capacity, antioxidant substance content, and antioxidant enzyme activity of nano-Se-treated peaches are higher, with potential storage resistance and health effects on human body. The mechanism of nano-Se affecting peach quality was analyzed by metabolomics and transcriptomics, which is a reference and guide for the research and application of nano-Se.

Quality changes of HHP orange juice during storage: Metabolomic data integration analyses.

High hydrostatic pressure (HHP) is a non-thermal method of sterilizing orange juice. However, knowledge of the quality variation during its storage is limited. This study aimed to comprehensively analyze metabolite variations during HHP orange juice storage using gas chromatography-mass spectrometer and liquid chromatography-mass spectrometry. Fifty-seven volatiles and 49 non-volatiles were identified. Partial least square analysis results showed that 21 days was a dividing point for metabolites highly varied. Results of relative odor activity value showed nonanal, methyl butanoate, and ethyl butanoate decreased after six days, which might reduce fruity flavor. After 21 days, over 60 % of metabolites such as linalool, α-pinene, and ascorbic acids decreased while α-terpineol and limonin increased, which would likely result in a change of coniferous, tarry, and bitter, as well as decreased organoleptic quality and antioxidative activities. This study provides a theoretical basis to optimize the shelf-life of HHP orange juice and advice for consumers' choices.

An integrated untargeted metabolomic approach reveals the quality characteristics of black soybeans from different geographical origins in China.

Black soybeans are extensively planted and consumed in China due to their high nutritional value and numerous health benefits. However, very few is known about the characteristic metabolites of black soybeans from different geographical origins in China. In the present study, 31 black soybean samples were collected from 11 main producing provinces in China. A combined metabolomics approach using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) and gas chromatography coupled to an Orbitrap mass analyzer (GC-orbitrap-MS) was performed for the first time to comprehensively investigate the metabolite variability among the black soybeans from different geographical origins. A total of 48 differential non-volatile metabolites and 14 differential volatile metabolites were identified based on orthogonal partial least squares discriminant analysis (OPLS-DA) coupled with analysis of variance (ANOVA). Higher procyanidin B1, procyanidin B2, epicatechin, malonylated isoflavones, and ß-pinene were observed in Gansu black soybeans. Guangxi black soybeans had higher amounts of linoleic acid and its oxidation products of hexanal and pentane. The black soybeans from Xinjiang and Yunnan were found to have higher delphinidin-derived anthocyanins, gamma-glutamyl peptides, and aromatic hydrocarbons. The characteristic metabolites of black soybeans from other geographical origins were also clarified. This study indicated that the integrated untargeted metabolomic approach can be a powerful tool to provide knowledge for developing specialty black soybeans.

Metabolomic profiles of not from concentrate orange juice after different sterilization treatments based on HS-SPME-GC-MS and UPLC-QTOF-MS.

Not from concentrate (NFC) orange juice is minimally processed, natural-appearing food, and has become more popular. Sterilization is an important stage for NFC orange juice production. Here we present a comprehensive analysis of the effect of sterilization on the metabolites of NFC orange juices, including three thermal (pasteurization, high-temperature short time, and ultra-high temperature) and one nonthermal (high hydrostatic pressure) method. A total of 108 metabolites, including 59 volatiles and 49 nonvolatiles were identified in orange juice. Of which, only butyl butanoate and 3-carene were detected in fresh orange juice. Sterilization significantly changed the metabolites of orange juice, and different methods caused various changes. Esters were downregulated by both thermal and nonthermal sterilization, while most flavonoids and terpenes were upregulated. With comparative analyses of three thermal sterilization, we found that high temperature and relatively short-time treatment preserved esters and ascorbic acid more effectively than low temperature and prolonged treatment. Aldehydes, however, were the opposite. Nonthermal sterilization is effective in preserving the metabolites of orange juice, especially for esters, terpenes, and flavonoids. In addition, 19 distinct metabolites were characterized between thermal and nonthermal samples by chemometrics analysis. These findings provide a new sight of the optimization of sterilization methods and references for different types of NFC orange juice identification. PRACTICAL APPLICATION: This study provides a reference for the optimization of sterilization methods and identification of HHP and thermal NFC orange juice and also benefits the purchase of consumers.

Hot Water Treatment Improves Date Drying and Maintains Phytochemicals and Fruit Quality Characteristics of Date Palm (Phoenix dactylifera).

Fresh date fruits (cvs. Hillawi and Khadrawi) were harvested at the khalal stage and treated with hot water treatment (HWT) for different time durations (control, HWT-1 min, HWT-3 min, HWT-5 min, and HWT-7 min) to investigate the physicochemical characteristics, phytochemical properties, and sensory attributes. The results revealed that both date cultivars took less time to reach the tamar stage in response to HWT-7 min compared to control. However, Hillawi date fruit showed a higher fruit ripening index (75%) at HWT-3 min, while Khadrawi fruit had a higher ripening index (80%) at HWT-5 min than untreated fruit (10%). Higher weight loss and lower moisture contents were observed in Hillawi (25%) and Khadrawi (20%) date fruit as the immersion period increased in both cultivars. Moreover, soluble solid content was higher in Hillawi (11.77° Brix) in response to HWT-3 min and Khadrawi (10.02° Brix) date fruit immersed in HWT-5 min in contrast with the control group, whereas significantly lower levels of titratable acidity and ascorbic acid content were observed in Hillawi (0.162%, 0.67 mg/100 g) and Khadrawi (0.206%, 0.73 mg/100 g) date fruit in response to HWT (HWT-1 min, HWT-3 min, HWT-5 min, and HWT-7 min) than untreated fruit. Furthermore, noticeably higher levels of reducing sugar (69.83%, 57.01%), total sugar (34.47%, 31.14%), glucose (36.84%, 29.42%), fructose (33.99%, 27.61%), and sucrose (3.16%, 1.33%) were found in hot water-treated Hillawi (immersed for 3-min) and Khadrawi (immersed for 5-min) date fruit, respectively. In addition, total phenolic content, total flavonoids, total antioxidants, and total tannins were substantially superior in date fruits subjected to HWT-3 min (in Hillawi, 128 mg GAE/100 g, 61.78%, 20.18 mg CEQ/100 g) and HWT-5 min (in Khadrawi, 139.43 mg GAE/100 g, 72.84%, and 18.48 mg CEQ/100 g) compared to control. Overall, sensory attributes were recorded to be higher in Hillawi and Khadrawi date fruit after treatment for 3 min and 5 min, respectively. Our findings suggest that HWT is a promising technique that can be adopted commercially to improve fruit ripening and preserved nutritional quality of dates after harvest.

Ultrasonic-microwave synergistic supramolecular solvent liquid-liquid microextraction of trace biogenic amines in fish and beer based on solidification of floating organic droplet.

It is important to detect the presence of biogenic amines (BAs) as indicators of food freshness. The purpose of this study was to develop a novel ultrasonic-microwave synergistic supramolecular solvent liquid-liquid microextraction based on solidification of floating organic droplet (UMS-SUPRAS-SFO-LLME) combined with high-performance liquid chromatography for the determination of BAs. The physical properties and microstructure of SUPRAS based on 1-dodecanol and tetrahydrofuran were studied, and the extraction conditions such as the SUPRAS volume, the UMS process, and the centrifugal conditions were optimized. The results for the extraction kinetics and thermodynamics showed that UMS-SUPRAS-SFO-LLME is a spontaneous, endothermic diffusion process. The linear ranges of this method are 0.1-2.0 × 105 ng·mL-1 (R2 > 0.994), the limits of detection are 4.0 × 10-3-6.0 × 10-2 ng·mL-1, and the recoveries were 96.28-103.15%. Compared with existing analysis methods, UMS-SUPRAS-SFO-LLME is a sensitive, green and economical sample pretreatment method for analyzing the enrichment of BAs in beer and fish.

Separation and purification of neohesperidin from the albedo of Citrus reticulata cv. Suavissima by combination of macroporous resin and high-speed counter-current chromatography.

In this article, a simple and efficient protocol for rapid preparation and separation of neohesperidin from the albedo of Citrus reticulata cv. Suavissima was established by the combination of macroporous resin column chromatography and high-speed counter-current chromatography (HSCCC). Six types of resin were investigated by adsorption and desorption tests, and D101 macroporous resin was selected for the first cleaning-up procedure, in which 55% aqueous ethanol was used to elute neohesperidin. After treatment with D101 resin, the neohesperidin purity increased 11.83-fold from 4.92% in the crude extract to 58.22% in the resin-refined sample, with a recovery of 68.97%. The resin-refined sample was directly subjected to HSCCC purification with a two-phase solvent system composed of ethyl acetate-n-butanol-water (4:1:5, v/v), and 23.6 mg neohesperidin with 97.47% purity was obtained from 60 mg sample in only one run. The recovery of neohesperidin in HSCCC separation procedure was 65.85%. The chemical structure of the purified neohesperidin was identified by both HPLC and LC-MS. The established purification process will be helpful for further characterization and utilization of Citrus neohesperidin.

Quantification and purification of mangiferin from Chinese Mango (Mangifera indica L.) cultivars and its protective effect on human umbilical vein endothelial cells under H(2)O(2)-induced stress.

Mangiferin is a natural xanthonoid with various biological activities. Quantification of mangiferin in fruit peel, pulp, and seed kernel was carried out in 11 Chinese mango (Mangifera indica L.) cultivars. The highest mangiferin content was found in the peel of Lvpimang (LPM) fruit (7.49 mg/g DW). Efficient purification of mangiferin from mango fruit peel was then established for the first time by combination of macroporous HPD100 resin chromatography with optimized high-speed counter-current chromatography (HSCCC). Purified mangiferin was identified by both HPLC and LC-MS, and it showed higher DPPH(•) free-radical scavenging capacities and ferric reducing ability of plasma (FRAP) than by l-ascorbic acid (Vc) or Trolox. In addition, it showed significant protective effects on human umbilical vein endothelial cells (HUVEC) under H(2)O(2)-induced stress. Cells treated with mangiferin resulted in significant enhanced cell survival under of H(2)O(2) stress. Therefore, mangiferin from mango fruit provides a promising perspective for the prevention of oxidative stress-associated diseases.