The Effect of Fermentation on the Chemical Constituents of Gastrodia Tuber Hallimasch Powder (GTHP) Estimated by UHPLC-Q-Orbitrap HRMS and HPLC.

OBJECTIVE: To compare the effect of fermentation on the chemical constituents of Gastrodia Tuder Halimasch Powder (GTHP), to establish its fingerprinting and multicomponent content determination, and to provide a basis for the processing, handling, and clinical application of this herb. METHODS: Ultra-high-performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to conduct a preliminary analysis of the chemical constituents in GTHP before and after fermentation. High-performance liquid chromatography (HPLC) was used to determine some major differential components of GTHP and establish fingerprints. Cluster analysis (CA), and principal component analysis (PCA) were employed for comprehensive evaluation. RESULTS: Seventy-nine compounds were identified, including flavonoids, organic acids, nucleosides, terpenoids, and others. The CA and PCA results showed that ten samples were divided into three groups. Through standard control and HPLC analysis, 10 compounds were identified from 22 peaks, namely uracil, guanosine, adenosine, 5-hydroxymethylfurfural (5-HMF), daidzin, genistin, glycitein, daidzein, genistein, and ergosterol. After fermentation, GTHP exhibited significantly higher contents of uracil, guanosine, adenosine, 5-hydroxymethylfurfural, and ergosterol and significantly lower genistein and daidzein contents. CONCLUSIONS: The UHPLC-Q-Orbitrap HRMS and HPLC methods can effectively identify a variety of chemical components before and after the fermentation of GTHP. This study provides a valuable reference for further research on the rational clinical application and quality control improvement of GTHP.

Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade.

Glyoxal oxidases, belonging to the group of copper radical oxidases (CROs), oxidize aldehydes to carboxylic acids, while reducing O2 to H2O2. Their activity on furan derivatives like 5-hydroxymethylfurfural (HMF) makes these enzymes promising biocatalysts for the environmentally friendly synthesis of the bioplastics precursor 2,5-furandicarboxylic acid (FDCA). However, glyoxal oxidases suffer from inactivation, which requires the identification of suitable redox activators for efficient substrate conversion. Furthermore, only a few glyoxal oxidases have been expressed and characterized so far. Here, we report on a new glyoxal oxidase from Trametes versicolor (TvGLOX) that was expressed at high levels in Pichia pastoris (reclassified as Komagataella phaffii). TvGLOX was found to catalyze the oxidation of aldehyde groups in glyoxylic acid, methyl glyoxal, HMF, 2,5-diformylfuran (DFF) and 5-formyl-2-furancarboxylic acid (FFCA), but barely accepted alcohol groups as in 5-hydroxymethyl-2-furancarboxylic acid (HMFCA), preventing formation of FDCA from HMF. Various redox activators were tested for TvGLOX reactivation during catalyzed reactions. Among them, a combination of horseradish peroxidase and its substrate 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS) most efficiently reactivated TvGLOX. Through continuous reactivation of TvGLOX in a two-enzyme system employing a recombinant Moesziomyces antarcticus aryl-alcohol oxidase (MaAAO) almost complete conversion of 8 mM HMF to FDCA was achieved within 24 h.

A chemometric study on the identification of 5-methylfurfural and 2-acetylfuran as particular volatile compounds of oxidized fish oil based on SHS-GC-IMS.

Fish oil develops particular off-odors, mainly fishy odor, from the oxidation of its characteristic fatty acids, docosahexaenoic (DHA) and eicosapentaenoic (EPA). Anchovy oil (AO) was taken as representative of fish oils. This was compared to three vegetable oils with different fatty acid compositions, i.e. camellia, sunflower and linseed oil, and differential volatile compounds were identified by static-headspace gas-chromatography ion-mobility-spectrometry (SHS-GC-IMS) and orthogonal partial-least-squares discriminant analysis (OPLS-DA) during oxidation at 60 °C. Three groups of differential volatile compounds detected at higher concentrations in the AO were screened out and two compounds, identified as 5-methylfurfural and 2-acetylfuran, were characteristic to the AO and not found in the vegetable oils. They were formed from both EPA and DHA, only present in the AO, and their formation mechanisms were proposed. The contents of 5-methylfurfural and 2-acetylfuran increased linearly with the oxidation time and consequently they could be used as oxidative markers of fish oils.

Soluble dietary fiber from tea residues with inhibitory effects against acrylamide and 5-hydroxymethylfurfural formation in biscuits: The role of bound polyphenols.

This study investigated the impact of soluble dietary fiber (SDF) from untreated (U-SDF), fermented (F-SDF) and high temperature cooked (H-SDF) from tea residues on formation of acrylamide (AA) and 5-hydroxymethylfurfural (5-HMF) in biscuits. Both 3% F-SDF and 2% H-SDF can simultaneously inhibit AA and 5-HMF and SDFs increased the types of volatile compounds in biscuits. After the determination of the bound polyphenol compositions in SDFs by LC-QTOF-MS/MS, six polyphenols with different structural characteristics were selected to explore their contributions on the inhibitory effect of SDFs and structure-inhibitory capacity relationships in the "glucose-asparagine-linoleic acid" model system. It showed that the inhibitory activities of those polyphenols were greatly affected by the number of hydroxyl groups and methoxy groups on the benzene ring. Almost all polyphenols were also found to scavenge hydroxyl radicals generated in reactions. Thus, this study suggests that the bound polyphenols of SDFs play a key role in the inhibition of AA and 5-HMF.

Application of acidic deep eutectic solvents in green extraction of 5-hydroxymethylfurfural.

The extraction of the 5-hydroxymethylfurfural (5-HMF), as building block for many applications, from aqueous solutions has been became an indispensable challenge. Consequently, this study investigated the extraction ability of acidic deep eutectic solvent-based aqueous two-phase system (ATPS), choline chloride as hydrogen bond acceptor (HBA) and lactic acid, oxalic acid and citric acid as hydrogen bond donor (HBD), countering polypropylene glycol 400 at T = 298.15 K. Two semi-empirical Zafarani-Moattar et al. and Merchuk equations were used to fit the measured binodal data. Further, the NRTL and UNIQUAC models were used for correlating of tie-line data. The consistency of the experimental tie-line data was determined by utilizing the Bachman-Brown and Hand correlations. Also, the performance of these ATPSs to partitioning of 5-HMF, were investigated by calculation of extraction efficiencies, EE% and partition coefficients, K. This strategy indicates DES-based ATPSs have the acceptable extraction efficiency of the 5-HMF in a single-step extraction. A greenness assessment tool, Analytical GREEnness metric (AGREE), was tested to evaluate of greenness of analytical protocol.

Cu-NPs@C Nanosheets Derived from a PVP-assisted 2D Cu-MOF with Renewable Ligand for High-Efficient Selective Hydrogenation of 5-Hydroxymethylfurfural.

5-Hydroxymethylfurfural (HMF) containing C=O, C-O, and furan ring functional groups is an important platform chemical derived from C6 sugars. The selective hydrogenation of C=O in HMF produces 2,5-dihydroxymethylfuran (DHMF), which is a potential sustainable substitute for petroleum-based building blocks. Here, 2,5-furandicarboxylic acid (H2 FDC), a promising sustainable alternative to terephthalic acid, was employed as a renewable ligand to synthesize a novel Cu metal-organic framework (Cu-FDC). With a polyvinyl pyrrolidone (PVP)-assisted approach, 2D Cu-FDC nano-lamellae of micrometer lateral dimensions and nanometer thickness could be obtained, which could be used as a precursor to fabricate 2D oxygen-rich carbon nanosheets embedded with Cu nanoparticles (denoted CFP-300) after a thermal treatment at 300 °C under N2 atmosphere. The synthesized CFP-300 exhibited excellent catalytic performance and stability for the selective hydrogenation of HMF to DHMF. These results demonstrated a sustainable route to synthesize efficient catalysts by employing metal-organic frameworks based on renewable ligands.

Low-temperature hydrothermal liquefaction of pomelo peel for production of 5-hydroxymethylfurfural-rich bio-oil using ionic liquid loaded ZSM-5.

Ionic liquid loaded ZSM-5 with high stability and catalytic performance was used for hydrothermal liquefaction (HTL) of pomelo peel for the first time. Bio-oil obtained at 200 °C had the highest yield (29.21 wt%) and high heating value (21.41 MJ/kg), with main constituents of 5-hydroxymethylfurfural (5-HMF, 50.10%), 3-Pyridinol (19.8%) and pentanoic acid (5.35%). The higher 5-hydroxymethylfurfural yield obtained using ionic liquid loaded ZSM-5 was further compared to other studies (0-50%). In comparison to high-temperature HTL, catalytic HTL with ionic liquid loaded ZSM-5 led to lower activation energy requirements (31.93 kJ·mol-1) for the conversion of glucose into 5-HMF. Additionally, the catalysts showed excellent recyclability, with 19.68 wt% of bio-oil containing 59.6% of light oil obtained after 5 cycles. Hence, this study presents a novel approach for the catalytic conversion of lignocellulosic biomass into 5-HMF-rich bio-oil for energy and green chemistry applications.

5-Hydroxymethylfurfural Exerts Negative Effects on Gastric Mucosal Epithelial Cells by Inducing Oxidative Stress, Apoptosis, and Tight Junction Disruption.

5-Hydroxymethylfurfural (5-HMF) is a processing byproduct present in foods that are consumed daily by humans, and the diet is the principal route for human exposure to it. However, its adverse effects on gastric epithelial cells are not fully understood. Based on the half inhibitory concentration value, concentrations of HMF of 2, 4, 8, and 16 mM were selected for this study. After 5-HMF treatment for 24 h, the number of living cells decreased to 89.61 ± 0.48, 77.30 ± 0.57, 58.75 ± 0.36, and 19.61 ± 0.40% of the control, respectively. Apoptosis activated through both the death receptor and mitochondrial pathways was confirmed to be the primary mode of HMF-induced cell death. Further analysis revealed that the reactive oxygen species (ROS) levels in GES-1 cells increased 1.7-6.5 fold after exposure to 5-HMF. Moreover, the inhibition of ROS by N-acetylcysteine blocked HMF-induced apoptosis and cell proliferation suppression, indicating that oxidative stress was important in HMF-induced apoptosis. Besides, after 5-HMF treatment, the gene expressions of occludin and ZO-1 were reduced by 1.1-3.4 fold and 2.0-9.4 fold, respectively. The cell surface morphology and tight junction-related protein expression analysis also revealed the destructive effect of 5-HMF on tight junction integrity. Our research highlights a potential mechanism of HMF-induced toxicity in GES-1 cells and provides additional information on the health risks of 5-HMF exposure to the human gastric epithelium.

Ambient-Temperature Reductive Amination of 5-Hydroxymethylfurfural Over Al2 O3 -Supported Carbon-Doped Nickel Catalyst.

An efficient catalytic system for the conversion of 5-hydroxymethylfurfural (HMF) into N-containing compounds over low-cost non-noble-metal catalysts is preferable, but it is challenging to reach high conversion and selectivity under mild conditions. Herein, an Al2 O3 -supported carbon-doped Ni catalyst was obtained via the direct pyrolysis-reduction of a mixture of Ni3 (BTC)2 ⋅ 12H2 O and Al2 O3 , generating stable Ni0 species due to the presence of carbon residue. A high yield of 96 % was observed in the reductive amination of HMF into 5-hydroxymethyl furfurylamine (HMFA) with ammonia and hydrogen at ambient temperature. The catalyst was recyclable and could be applied to the ambient-temperature synthesis of HMF-based secondary/tertiary amines and other biomass-derived amines from the carbonyl compounds. The significant performance was attributable to the synergistic effect of Ni0 species and acidic property of the support Al2 O3 , which promoted the selective ammonolysis of the imine intermediate while inhibiting the potential side reaction of over-hydrogenation.

Gallic Acid Mitigates 5-Hydroxymethylfurfural Formation while Enhancing or Preserving Browning and Antioxidant Activity Development in Glucose/Arginine and Sucrose/Arginine Maillard Model Systems.

The current trend of lowering 5-hydroxymethylfurfural (5-HMF) dietary exposure is challenging since its formation is parallel with the development of food color, flavor and aroma. We aimed to investigate the effect of gallic acid (GA) addition on 5-HMF formation, color development and antioxidant activity (AA) in a series of Maillard Reaction (MR) model systems. The effects of GA addition on browning and AA development were not uniform for all model systems, but always occurred in the same direction, indicating that these phenomena were interconnected. GA mitigated 5-HMF development in four of the nine tested systems, possibly by preventing the oxidation of MR intermediates. Correlation analysis indicated that when GA addition mitigated 5-HMF formation, browning was either promoted or not affected. The proposed strategy was effective for glucose/arginine and sucrose/arginine systems, since GA mitigated 5-HMF formation (49% and 54%, respectively) in addition to increasing color development and antioxidant activity.

A quick and simple paper-based method for detection of furfural and 5-hydroxymethylfurfural in beverages and fruit juices.

Improper storage and transportation of non-alcoholic beverages can, over longer periods, induce Maillard reaction, degrading nutritional components and generating genotoxic and carcinogenic by-products such as furfural and 5-hydroxymethylfurfural (HMF), rendering products unsafe for human consumption. Here, we describe a rapid quantitative solution-based method and test-strips for detection of furfural and HMF. The standard spectroscopic method indicated an LOD of 0.006 ±â€¯0.003% (v/v) and 0.005 ±â€¯0.002% (v/v) for furfural and HMF, respectively in fruit juice samples. The novel chromogenic test-strip has sensitivity of 0.008% (v/v) and 0.004% (v/v) for furfural and HMF, respectively in the same samples of fruit juice. Thus, the developed method and test-strips were specific for furfural and HMF and can be used to help ensure food safety and quality in various industrial applications.

Tandem Thio-Michael Addition/Remote Lactone Activation of 5-Hydroxymethylfurfural-Derived δ-Lactone-Fused Cyclopentenones.

The creation of structurally diverse chemical entities from fairly simple biorefinery products remains a challenge. In this work 5-hydroxymethylfurfural (HMF) was identified as a key synthon for preparing highly complex cyclopentenones (CP) via tandem 1,4-addition/elimination/remote lactone activation to external O- and N-nucleophiles in δ-lactone-fused-CPs hotspots. This scaffold was also reactive enough to be incorporated into model cysteine-peptides in low concentrations, paving the way to a potential translation generating complexity in the synthesis of small peptides. The new enones also exhibited activity against intraerythrocytic Plasmodium falciparum (IC50 =1.32 µm).

5-Hydroxymethyl-2-furaldehyde induces developmental toxicology and decreases bone mineralization in zebrafish larvae.

In this study, we aimed to assess the developmental toxicity and effects of 5-HMF in zebrafish as a model organism for toxicology studies. To this end, we treated zebrafish embryos with 1-100 µg/ml 5-HMF and observed bone staining, gene expression, and reactive oxygen species levels in order to investigate the toxicological effects of 5-HMF. The results showed that high concentrations of 5-HMF caused increased mortality and deformity rates in zebrafish larvae, inhibited cartilage development, reduced bone mineralization, increased reactive oxygen species levels, and disrupted the expression of genes related to bone development and reactive oxygen species enzyme activity. The antioxidant N-acetyl-l-cysteine partially rescued the toxicological effects caused by the high concentrations of 5-HMF. Overall, these findings showed that high concentrations of 5-HMF induce reactive oxygen species production, leading to developmental toxicity and decreased bone mineralization. Our results provide a reference for understanding the toxic effects of 5-HMF.

Coupling a recombinant oxidase to catalase through specific noncovalent interaction to improve the oxidation of 5-hydroxymethylfurfural to 2,5-furandicarboxylic acid.

5-Hydroxymethylfurfural oxidase (HMFO) can catalyze both hydroxyl and aldehyde oxidations. It catalyzes 5-hydroxymethylfurfural into 2,5-furandicarboxylic acid. However, the application of HMFO encountered two problems: the expressed HMFO in Escherichia coli. is largely in the form of inclusion bodies, and the by-product of H2O2 has a negative effect on HMFO stability. To solve these problems, recombinant HMFO was generated by fusing the C-terminus to an elastin-like polypeptide (ELP). ELP-HMFO can be expressed with significantly reduced inclusion bodies. ELP-HMFO exhibited improved stability and tolerance toward H2O2. Further recombination is carried out by fusing the N-terminus of HMFO to a glutamic acid-rich leucine zipper motif (ZE). Similarly, recombinant catalase (CAT) is generated by fusing the N-terminus to ELP and fusing the C-terminus to an arginine-rich leucine zipper motif (ZR). ELP-HMFO-ZE can interact specifically with ZR-CAT-ELP, ascribing to the coiled-coil association of ZE and ZR. ELP-HMFO-ZE#ZR-CAT-ELP coordinates the respective catalytic activities of the two enzymes. ELP-HMFO-ZE catalyzes the oxidation of HMF, and the generated hydrogen peroxide is decomposed by ZR-CAT-ELP into H2O and oxygen. During the oxidation of HMF, the cofactor FAD of HMFO is reduced, and molecular oxygen is needed to reoxidize the reduced FAD. The evolved oxygen from the decomposing of H2O2 can just meet the requirement, which can be diffused efficiently from ZR-CAT-ELP to ELP-HMFO-ZE due to the short distance between the two enzymes.

Headspace Solid-Phase Microextraction Analysis of Volatile Components in Peanut Oil.

Peanut oil is favored by consumers due to its rich nutritional value and unique flavor. This study used headspace solid-phase microextraction (HS-SPME) combined with gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) to examine the differences in the peanut oil aroma on the basis of variety, roasting temperatures, and pressing components. The results revealed that the optimal conditions for extracting peanut oil were achieved through the use of 50/30 µm DVB/CAR/PDMS fibers at 60 °C for 50 min. The primary compounds present in peanut oil were pyrazines. When peanuts were roasted, the temperature raised from 120 °C to 140 °C and the content of aldehydes in peanut oil increased; however, the content of aldehydes in No. 9 oil at 160 °C decreased. The components of peanut shell oil varied depending on the peanut variety. The most marked difference was observed in terms of the main compound at the two roasting temperatures. This compound was a pyrazine, and the content increased with the roasting temperature in hekei oils. When the roasting temperature was lower, No. 9 oil contained more fatty acid oxidation products such as hexanal, heptanal, and nonanal. When the roasting temperature increased, No. 9 oil contained more furfural and 5-methylfurfural. Heren oil was easier to oxidize and produced nonanal that possessed a fatty aroma.

Intensive pulsed light pretreatment combined with controlled temperature and humidity for convection drying to reduce browning and improve quality of dried shiitake mushrooms.

BACKGROUND: The change of surface color caused by browning during the drying process of shiitake mushrooms seriously affects its market circulation. Intensive pulsed light (IPL) as a non-heat-treatment method can reduce enzyme activity by changing the enzyme structure. Therefore, in this study, the use of IPL pretreatment before drying was aimed to reduce the adverse reactions caused by the browning reaction during the drying processing of shiitake mushrooms. RESULTS: Shiitake mushrooms pretreated with 25 pulses of IPL energy of 400 J reduced the initial polyphenol oxidase enzyme activity, the browning index, and browning degree values by 42.83%, 43.02%, and 47.54% respectively. The IPL pretreatment enhanced the polysaccharides and reducing sugars contents and it reduced 5-hydroxymethylfurfural generation in the dried shiitake mushrooms. The pretreatment also improved the surface color, the antioxidant activity, and retained the umami taste characteristics in the dried shiitake mushroom. CONCLUSION: The IPL pretreatment combined with controlled temperature and humidity for convection drying could be a suitable method to improve the quality of dried shiitake mushrooms. Therefore, this study provides a new pretreatment method for materials that are prone to browning during drying. © 2021 Society of Chemical Industry.

Comprehensive Study on the Acrylamide Content of High Thermally Processed Foods.

Acrylamide (AA) formation in starch-based processed foods at elevated temperatures is a serious health issue as it is a toxic and carcinogenic substance. However, the formation of more AA entangles with modern-day fast food industries, and a considerable amount of this ingredient is being consumed by fast food eaters inadvertently throughout the world. This article reviews the factors responsible for AA formation pathways, investigation techniques of AA, toxicity, and health-related issues followed by mitigation methods that have been studied in the past few decades comprehensively. Predominantly, AA and hydroxymethylfurfural (HMF) are produced via the Maillard reaction and can be highlighted as the major heat-induced toxins formulated in bread and bakery products. Epidemiological studies have shown that there is a strong relationship between AA accumulation in the body and the increased risk of cancers. The scientific community is still in a dearth of technology in producing AA-free starch-protein-fat-based thermally processed food products. Therefore, this paper may facilitate the food scientists to their endeavor in developing mitigation techniques pertaining to the formation of AA and HMF in baked foods in the future.

5-Hydroxymethylfurfural and α-ketoglutaric acid supplementation increases oxygen saturation during prolonged exercise in normobaric hypoxia.

This double-blinded, randomized and placebo-controlled, crossover study investigated whether α-ketoglutaric-acid (α-KG) and 5-hydroxymethylfurfural (5-HMF) supplementation improves exercise performance in hypoxia and affects physiological responses during the exercise task. Eight moderately trained male participants (age: 25.3 ± 2.0 y, VO2max: 48.0 ± 8.3 ml/min/kg) performed an incremental exercise test to exhaustion in normoxia and two 2-hour cycle time trial (TT) tests in hypoxia (3,500 m) each separated by 1-week. Prior to the TT, participants supplemented with either α-KG and 5-HMF or placebo (random order). Supplementation did not improve TT performance at altitude and did not affect heart rate, effort perception and oxidative stress levels (p > 0.05). Oxygen saturation (SpO2) was enhanced during the α-KG and 5-HMF supplementation trial (79.5 ± 3.3 vs. 78.2 ± 3.7%, p = 0.026). Even though TT performance was unaffected, the enhanced SpO2 - possibly originated from changed O2-affinity - deserves further consideration as the exercise performance decline at altitude is strongly linked to the SpO2 decline. The inclusion of moderately fit participants, not specifically cycle trained, might have prevented any visible performance enhancement.

Path analysis of non-enzymatic browning in Dongbei Suancai during storage caused by different fermentation conditions.

A non-enzymatic browning reaction occurs easily in Dongbei Suancai (DS) during storage. Using the method of path analysis, the changes in contents of VC (ascorbic acid), polyphenol, reducing sugar, amino nitrogen, and 5-HMF (5-hydroxymethyl furfural) were investigated to analyze the direct pathways and determinants of browning caused by pre-production blanching at 100 °C (R-100), salt-addition increased from 2% to 6% (Y-6) and fermentation time extended from 30 d to 40 d (T-40), respectively. The results showed that R-100 could delay the browning by inhibiting two direct pathways of oxidative decomposition of VC and oxidation-polymerization of polyphenols, but T-40 could lead to an increase in degree of browning for which primary determinant was the interaction between polyphenol and reducing sugar, while Y-6 had no obvious effect on browning pathway and determinants. R-100 was thus deemed to be a good measure with inhibiting non-enzymatic browning in DS during storage.

A Novel Chinese Honey from Amorpha fruticosa L.: Nutritional Composition and Antioxidant Capacity In Vitro.

False indigo (Amorpha fruticosa L., A. fruticosa) is the preferred tree indigenous for windbreak and sand control in Northwest China, while information on nutritional and bioactive characteristics of its honey is rare. Herein, 12 honey of Amorpha fruticosa L. (AFH) were sampled in Northwest China and the nutritional composition was determined. Sixteen mineral element and ten dominant polyphenols content were identified and quantified by ICP-MS (Inductively coupled plasma mass spectrometry) and HPLC-QTOF-MS (High performance liquid chromatography-Quadrupole time-of-flight mass spectrometry), respectively. Moreover, AFH demonstrated high levels of DPPH (1,1-Diphenyl-2-picrylhydrazyl) radical scavenging activity (IC50 100.41 ± 15.35 mg/mL), ferric reducing antioxidant power (2.04 ± 0.29 µmol FeSO4·7H2O/g), and ferrous ion-chelating activity (82.56 ± 16.01 mg Na2EDTA/kg), which were significantly associated with total phenolic contents (270.07 ± 27.15 mg GA/kg) and ascorbic acid contents (213.69 ± 27.87 mg/kg). The cell model verified that AFH exhibited dose-dependent preventive effects on pBR322 plasmid DNA and mouse lymphocyte DNA damage in response to oxidative stress. Taken together, our findings provide evidence for the future application of AFH as a potential antioxidant dietary in food industry.