Xiao-mei Sha

Improved Glycation after Ultrasonic Pretreatment Revealed by High-Performance Liquid Chromatography–Linear Ion Trap/Orbitrap High-Resolution Mass Spectrometry

The glycation extent of bovine serum albumin (BSA) before and after ultrasonication was evaluated by MALDI-TOF and Orbitrap mass spectrometry. Ultrasonic pretreatment significantly improved the incorporation of galactose to BSA. Prior to ultrasonic pretreatment, only 12 sites (11 lysines and 1 arginine) were glycated, whereas the number of glycation sites was increased to 42, including 39 lysines and 3 arginines, after treatment. Average degree of substitution per peptide molecule of BSA (DSP) was used to evaluate the glycation level for each glycation site. The ultrasonic pretreatment significantly improved the DSP value of all glycation sites. The prevalently promoted glycation by ultrasonic pretreatment suggests that ultrasonication improves glycation through altering the structure of BSA throughout all three domains. An ultrahigh-resolution linear ion trap Orbitrap mass spectrometer facilitates unambiguous localization of glycation sites, allowing an in-depth analysis of the nature and extent of protein glycation at the molecular level. High-intensity ultrasonication can greatly improve protein glycation and, therefore, opens new routes to modify the functionality of proteins in a positive way.

Gelatin Quantification by Oxygen-18 Labeling and Liquid Chromatography–High-Resolution Mass Spectrometry

Combined with high-performance liquid chromatography (HPLC) and linear-ion trap/Orbitrap high-resolution mass spectrometry, trypsin-catalyzed (16)O-to-(18)O exchange was used to establish an accurate quantitative method for bovine or porcine gelatin. The sophisticated modifications for these two mammalian gelatins were unambiguously identified by accurate mass and tandem mass spectrometry. Eighteen marker peptides were successfully identified for the bovine and porcine gelatin, respectively. The gelatins were subjected to (18)O or (16)O labeling in the presence of trypsin and mixed together in various ratios for quantification. All of the (18)O-labeled peptides were also confirmed by accurate mass and tandem mass spectrometry. The 10 marker peptides with the strongest signals were chosen to calculate the average ratios of (18)O-labeled and (16)O-labeled gelatin. The measured ratios of (18)O-labeled and (16)O-labeled peptides were very close to the mixing ratios of 20:1, 5:1, 1:1, and 1:5 with low standard deviation values. The samples with a mixing ratio of 1:1 (18)O-labeled and (16)O-labeled peptides were determined to 1.00 and 0.99 with standard deviations of 0.02 and 0.04 for bovine and porcine gelatins, respectively, indicating the high accuracy of this method. Trypsin-catalyzed (18)O labeling was proved to be an excellent internal calibrant for gelatins. When combined with HPLC and high-resolution mass spectrometry, it is an accurate and sensitive quantitative method for gelatin in the food industry.

Rheological behavior, emulsifying properties and structural characterization of phosphorylated fish gelatin

Rheological, microstructural and emulsifying properties of fish gelatin phosphorylated using sodium trimetaphosphate (STMP) were studied. Phosphorylation was carried out at 50 °C for 0, 0.5, 1 or 2 h. Rheological behaviors indicated that phosphorylation decreased gelation rate constant (kgel) and apparent viscosity of gelatin solutions. Phosphorylation time was inversely proportional to tan δ; gelling and melting points of fish gelatin gels; however gel properties could be improved by short time of phosphorylation. Scanning electron microscopy and atomic force microscopy revealed that longer time of phosphorylation resulted in looser gel network with more aggregation. Longer phosphorylation time could stabilize fish gelatin emulsions, and endowed emulsions with smaller particle size and lower coefficient viscosity, but higher ζ-potential values. These results suggested that phosphorylation could be applied to obtain fish gelatin with varying functional properties suitable for numerous industrial applications.

The Reduction in the IgE-Binding Ability of β-Lactoglobulin by Dynamic High-Pressure Microfluidization Coupled with Glycation Treatment Revealed by High-Resolution Mass Spectrometry

Our previous study indicated that pretreatment by dynamic high-pressure microfluidization (DHPM) and glycation with galactose was a promising method for decreasing the immunoglobulin E (IgE)-binding ability of β-lactoglobulin (β-LG). In this work, the conformational alteration of β-LG subjected to DHPM and glycation treatment was investigated in relation to IgE-binding ability by orbitrap mass spectrometry. After DHPM pretreatment, lower IgE-binding ability of glycated β-LG was observed with increasing pressures. Prior to DHPM pretreatment, 11 glycated sites were identified, while the number of glycation sites was increased to 12 after pretreatment. However, there was no significant difference of the glycation sites at the pressures of 50, 100, and 200 MPa, respectively. Average degree of substitution per peptide molecule of β-LG (DSP) was investigated to assess the degree of glycation per glycation site. All of the samples pretreated by DHPM exhibited a higher glycation level than those without DHPM pretreatment. The shielding effects of epitopes owing to glycation contributed to the reduction of IgE-binding capacity. Orbitrap mass spectrometry could provide a comprehensive understanding of the nature of protein glycation.

Jackfruit (Artocarpus heterophyllus Lam.) peel: A better source of antioxidants and a-glucosidase inhibitors than pulp, flake and seed, and phytochemical profile by HPLC-QTOF-MS/MS

Jackfruit (Artocarpus heterophyllus Lam.) peel is an underutilized by-product in both, the production and processing of jackfruit. This research compared the antioxidant and hypoglycemic potential of jackfruit peel with jackfruit pulp, flake and seed for the first time. The phytochemical profile of peel extract was characterized with HPLC-QTOF-MS/MS. Results revealed that peel extract exhibited the highest total phenolic and total flavonoid content, and the phenolics was 4.65, 4.12 and 4.95 times higher than that of pulp, flake and seed extract, respectively. The strongest DPPH and ABTS+ scavenging ability, α-glucosidase inhibition were also found in peel extract, and the α-glucosidase inhibition was about 11.8-fold of that of acarbose. The HPLC-QTOF-MS/MS analysis led to the tentative identification of 53 compounds, prenylflavonoids, hydroxycinnamic acids and glycosides are the predominant bioactive compounds. Above results reveal promising potential of jackfruit peel as a new source of natural antioxidants and hypoglycemic agents.

Rheological and structural properties of fish scales gelatin: Effects of conventional and ultrasound-assisted extraction

ABSTRACT In this study, the rheological and structural properties of extracted fish scales gelatin by conventional water bath (CWB) and ultrasound-assisted water bath (UWB) at different temperatures (60°C, 70°C and 80°C) were studied. Rheological behaviours showed that the extracted fish scales gelatin by UWB at 60°C possessed the highest storage modulus (5000 Pa), gelation point (22.94°C), melting point (29.54°C), and apparent viscosity than the others. X-ray showed that high temperature and ultrasound reduced the triple-helix content of fish scales gelatin along with decreasing gelling properties. Atomic force microscopy showed that the largest aggregates could be observed in the surface of UWB60. Therefore, UWB proved to be a promising technology in the extraction of high-quality fish gelatin.

The effect of ginger and garlic addition during cooking on the volatile profile of grass carp (Ctenopharyngodon idella) soup

Ginger and garlic have long been used in Asian countries to enhance the flavor and to neutralize any unpleasant odors present in fish soup. The purpose of this study was to evaluate the change in the amount of volatile components present in fish soup compared to boiled water solutions of ginger and garlic. The fish soup was prepared by boiling oil-fried grass carp (Ctenopharyngodon idella) with or without ginger and/or garlic. Generally, boiling garlic and ginger in water led to a decrease in the amount of the principal volatile constituents of these spices, together with the formation of some new volatiles such as pentanal, hexanal, and nonanal. The results showed that 16 terpenes present in raw ginger, predominantly camphene, β-phellandrene, β-citral, α-zingiberene, and (E)-neral, were detected in fish soup with added ginger and thus remained in the solution even after boiling. Similarly, 2-propen-1-ol and three sulfur compounds (allyl sulfide, diallyl disulfide, and diallyl trisulfide) present in raw garlic, were present in trace amounts in the boiled garlic solution, but were present in considerably larger amounts in the boiled fish solution with garlic or garlic plus ginger. In conclusion, the effect of adding spices on the volatile profile of grass carp soup can be attributed to the dissolution of flavor volatiles mainly derived from raw spices into the solution, with few additional volatiles being formed during boiling. In addition, boiling previously fried grass carp with spices led to enhanced volatile levels compared to boiled spice solutions.

Gelation kinetics and characterization of enzymatically enhanced fish scale gelatin–pectin coacervate

BACKGROUND Protein-polysaccharide complex coacervations have been considered extensively for the development of functional foods. The main problem of the complex coacervates is that they are highly unstable under different conditions and that cross-linking is necessary to stabilize them. In this study, the effects of pectin at different concentrations on the gel and structural properties of fish scale gelatin (FSG)-high methoxyl citrus pectin (HMP) coacervate enhanced by microbial transglutaminase (MTGase) were studied. RESULTS The gelation rates and gel strength of the MTGase-enhanced FSG-HMP coacervate gels decreased with increasing HMP concentration. However, the enhanced coacervate gels exhibited better thermal behavior and mechanical properties compared with the original gels. Also, TG-P8 exhibited the highest melting point (27.15 ± 0.12 °C), gelation point (15.65 ± 0.01 °C) and stress (15.36 ± 0.48 kPa) as HMP was 8 g kg-1 . Particle size distribution, fluorescence emission and UV absorbance spectra indicated that MTGase and HMP could make FSG form large aggregates. Moreover, confocal laser scanning microscopy of treated coacervate gels showed a continuous protein phase at low HMP concentrations. CONCLUSION FSG and HMP could form soluble coacervate, and MTGase could improve the thermal and mechanical properties of coacervate gels.

The Mechanism of Decreased IgG/IgE-Binding of Ovalbumin by Preheating Treatment Combined with Glycation Identified by Liquid Chromatography and High-Resolution Mass Spectrometry

Ovalbumin is one of the most important sensitizing ingredients in allergens of egg albumin, which restricts the application of egg in the field of food processing. Previous research has indicated that glycation could cause the protein to partially expand, which may bring about the destruction of the structural IgG and IgE epitopes and induce the decline of the IgG- and IgE-binding ability of ovalbumin. In this research, the effect of a preheating treatment integrated with glycation on the IgG- and IgE-binding capability and the conformation changes of ovalbumin was studied by detecting the glycated sites and the values of degree of substitution per peptide (DSP) by liquid chromatography and high-resolution mass spectrometry (LC-HRMS). Interestingly, we found that a glycation site (K227) attached by two ribose molecules was detected in glycated ovalbumin with preheating treatment. In addition, a new glycation site (K323) appeared in G-60. The results displayed that preheating treament could strengthen the changes in the secondary and tertiary structure of ovalbumin by enhancing glycation and further reduce the IgG/IgE-binding ability by integrating with glycation because of the cover of IgG and IgE epitopes. Therefore, preheating treatment integrated with glycation may offer a way for ovalbumin to reduce sensitization.

Ultrasonic Pretreatment Combined with Dry-State Glycation Reduced the Immunoglobulin E/Immunoglobulin G-Binding Ability of α-Lactalbumin Revealed by High-Resolution Mass Spectrometry

Bovine α-lactalbumin (α-LA) is one of major food allergens in cows milk. The present work sought to research the effects of ultrasonic pretreatment combined with dry heating-induced glycation between α-LA and galactose on the immunoglobulin E (IgE)/immunoglobulin G (IgG)-binding ability and glycation extent of α-LA, determined by inhibition enzyme-linked immunosorbent assay and high-resolution mass spectrometry, respectively. The IgE/IgG-binding ability of glycated α-LA was significantly decreased as a result of ultrasonic pretreatment, while the average molecular weight, incorporation ratio (IR) value, location and number of glycation sites, and degree of substitution per peptide (DSP) value were elevated. When the mixtures of α-LA and galactose were pretreated by ultrasonication at 150 W/cm2, glycated α-LA possesses seven glycation sites, the highest IR and DSP values, and the lowest IgE/IgG-binding ability. Therefore, the decrease in the IgE/IgG-binding ability of α-LA depends upon not only the shielding effect of the linear epitope found to be caused by the glycation of K13, K16, K58, K93, and K98 sites but also the intensified glycation extent, which reflected in the increase of the IR value, the number of glycation sites, and the DSP value. Moreover, allergenic proteins and monosaccharides pretreated by ultrasonication and then followed by dry-state glycation were revealed as a promising way of achieving lower allergenicity of proteins in food processing.