Formation mechanism of blue pigment in boiled lotus rhizome discs: Insight into the chelation of polyphenols and iron.

Boiled lotus rhizome discs (BLRDs), as common processed products of lotus rhizome, have gained increasing attention from consumers and food manufacturers. However, the blue pigment formed during boiling affects its appearance and reduces the appetite of BLRDs. In this study, the effects of polyphenols and iron contents on blue pigment formation in BLRDs in different regions and months were investigated. Results revealed that blue variation was more serious in March and April of the second year in Wuhan, and polyphenols and iron contents in these two months were significantly higher than those in other months. Then, UPLC and UV-Vis analysis showed that polyphenols causing the formation of blue pigment in BLRDs were L-dopa, gallocatechin, catechin, epigallocatechin, chlorogenic acid and epicatechin, among which L-dopa (52.450 mg/100 g in fresh lotus rhizome (FLR)) and gallocatechin (36.210 mg/100 g in FLR) possessed the greatest effect. Moreover, the ESI-Q-TOF-MS analysis of L-dopa-iron chelate and gallocatechin-iron chelate suggested that the blue pigment of BLRDs was mainly in the form of bis-complexes under boiling conditions. The study on formation mechanism of blue pigment in BLRDs can provide a reference for lotus rhizome processing.

Structure, spectral properties and antioxidant activity of melanoidins extracted from high temperature sterilized lotus rhizome juice.

Melanoidins are complex macromolecular compounds closely associated with the browning phenomenon in high-temperature sterilized lotus rhizome juice (HTSL). This study aimed to preliminarily investigate the structural properties of melanoidins extracted from HTSL. Results showed that the average molecular weight of HTSL melanoidins ranged from 1.48 to 41.40 kDa. Medium and high molecular weight melanoidins were the main contributors to the brown color of HTSL. Sugars, proteins, and phenolics were present in HTSL, among which sugar was the most abundant, with glucose being the predominant monosaccharide in acid degradation products of melanoidins. Through fluorescence and ultraviolet spectral analysis, we found that the melanoidins contained carboxyl and carbonyl compounds, as well as furan and pyran heterocyclic compounds. The infrared spectra and nuclear magnetic resonance spectra revealed a prominent sugar absorption peak, indicating that sugar was the main component of the melanoidins of HTSL. Furthermore, in vitro antioxidant experiments showed that the antioxidant activity of melanoidins was significantly positively correlated with phenolic compounds. Our results indicated that there were differences in the structural properties of melanoidins fractions with different molecular weights. MW-H fraction significantly impacted the color and antioxidant activity of HTSL.

Ultrasound-assisted extraction of polyphenols from lotus rhizome epidermis by alcohol/salt-based aqueous two-phase system: Optimization, extraction mechanism and antioxidant activities.

In this study, ultrasonic-assisted (UA) alcohol/salt-based aqueous two-phase system (ATPS) method was constructed to extract lotus rhizome epidermis (LRE) polyphenols. The extraction conditions were optimized as salt concentration 26.75 %, ethanol concentration 25.45 %, ultrasonic power 487 W and liquid-solid ratio 35.33 mL/g by comparing response surface methodology (RSM) and artificial neural network (ANN) models. Then, l-dopa (2.35 ± 0.036 mg/g dw), gallocatechin (1.66 ± 0.0035 mg/g dw) and epigallocatechin (1.37 ± 0.0035 mg/g dw) were determined as major polyphenols in LRE by using UA-ATPS method. Moreover, study showed that ultrasound, van der Waals force, hydrogen bond and salting out could accelerate the mass transfer and extraction of polyphenols in LRE cells. The high-pressure cavity and collapse effect of ultrasound could also accelerate the extraction of polyphenols. In vitro antioxidant experiments showed that LRE polyphenols have good antioxidant ability. In sum, this study developed a green and efficient extraction method to enhance the profitability of LRE in food and medicine industries.

Study on the formation mechanism of blackening in damaged lotus rhizome epidermis: Effects of polyphenols and iron.

Lotus rhizome is an important aquatic vegetable, but the blackening of lotus rhizome epidermis (LRE) seriously affects its appearance and quality, which makes lotus rhizome products unmarketable. In this study, the effects of polyphenols and iron on the LRE color were studied to explore the possible mechanism of LRE blackening. Results indicated that the measurable total phenols contents in the mud treatment (MT) group were significantly reduced, and the total iron contents were significantly increased compared with the bruised treatment group (p < 0.05). The high-performance liquid chromatography results showed that the main polyphenols in LRE were dopa, gallocatechin, and catechin, as well as a small amount of catechol, epicatechin, proanthocyanidin B2, and proanthocyanidin C1. Moreover, the results of color difference and ultraviolet adsorption spectroscopy showed that there were obviously black or brown-gray of dopa (525 nm), gallocatechin (504.5 nm), and catechin (550 and 504.5 nm) with FeCl2. The simulated system treatment of LRE further confirmed that the chromaticity effect of dopa and iron in bruised LRE was similar to that of the MT group, whereas 1% (w/w) ascorbic acid, 2% (w/w) EDTA-2Na, or 3% (w/w) citric acid could solely prohibit the blackening. This suggested that the dopa in LRE and FeCl2 in mud may mainly combine into [2(DOPA-2H+)+Fe3+]- through non-covalent interaction, which leads to the blackening of bruised LRE under neutral conditions. These results can guide the storage of lotus rhizomes and improve the development of the lotus rhizome industry.

Mechanism for gel formation of pectin from mealy and crisp lotus rhizome induced by Na+ and D-glucono-d-lactone.

Lotus rhizome residue, a cell wall material produced during the production of lotus rhizome starch, has long been underutilized. This study aims to extract pectin-rich polysaccharides from the cell wall of lotus rhizome and investigate their gelation mechanism in order to improve their industrial applicability. The results indicated that both CP and MP (pectin extracted from crisp and mealy lotus rhizome) exhibited a highly linear low methoxyl pectin structure, with the primary linkage mode being →4)-GalpA-(1→. The pectin chains in MP were found to be more flexible than those in CP. Then the impact of Na+, D-glucono-d-lactone (GDL), urea, sodium dodecyl sulfate (SDS), either individually or in combination, on the rheological characteristics of gels was evaluated. The results indicated that gels induced by GDL exhibited favorable thermoreversible properties, whereas the thermoreversibility of Na+-induced gels is poor. In addition to hydrogen bonding and ionic interactions, hydrophobic interactions also play a significant role in the formation of pectin gels. This study offers theoretical guidance and methodologies to improve the utilization rate of lotus rhizome starch processing by-products, while also provides novel insights into the correlation between LMP structure and gelation mechanism.

Analysis of pectin-cellulose interaction in cell wall of lotus rhizome with assistance of ball-milling and galactosidase.

The current study sought to depict the structural feature of polysaccharides extracted from Na2CO3 unextractable fraction (LUN) of lotus rhizome using galactosidase with assistance of ball milling. The extracted polysaccharides were a complex of cellulose microfibrils and the RG-I structural domain of pectin, and the top three monosaccharides were glucose, galactose and galactose uronic acid, which allowed to tune the properties of the enzyme-hydrolyzed polysaccharide from LUN after 15 and 45 min of ball milling. The data of XRD revealed that pectin has a masking effect on the diffraction peaks of cellulose components. The removing of the polysaccharides could increase the degree of crystallinity and the pectin-cellulose interaction mainly occured through the galactan side chain was speculated. Textural characterization by SEM exhibited a cross-linked rod-like structure, which is similar to the structure of cellulose microfibrils. The morphological analysis of AFM revealed that L15-P (enzyme-hydrolyzed polysaccharide from LUN after 15 min of ball milling) contained relatively ordered and uniform network structures. Overall, the present study provides an important insight into cell wall of lotus rhizome matrix polysaccharide.

Acetic Acid Immersion Alleviates the Softening of Cooked Sagittaria sagittifolia L. Slices by Affecting Cell Wall Polysaccharides.

This study investigated the mechanism for acetic acid pretreatment to improve cell wall integrity and thereby enhance the hardness of cooked Sagittaria sagittifolia L. slices by affecting polysaccharides in the cell wall. Distilled water immersion and 0.6% acetic acid immersion (the solid/liquid ratio is 1:10) for 15 h at room temperature could result in the conversion of pectin through different reactions during thermal processing. Combined in situ and in vitro analysis demonstrated that acetic acid pretreatment could promote the interaction of cellulose microfiber or hemicellulose with RG-Ⅰ side chains during thermal processing of S. sagittifolia L. slices, promote the entanglement between linear pectin molecules and make hemicellulose show a lower molecular weight under cooking, making it easy to firmly bind to pectin, which resulted in texture changes. The findings may help improve the texture of thermally processed vegetables and fruits and deep processing of starchy vegetables.

Effects of different concentrations of ascorbic acid on the stability of (+) - Catechin under enzymatic conditions.

A series of incubation systems of (+) - catechin (Cat), ascorbic acid (AA) and polyphenol oxidase (PPO) of lotus rhizome at 40 °C were performed to investigate the effect and oxidation pathway of AA on the stability of Cat. The results showed that after the enzymatic or non-enzymatic oxidation of Cat, the products of the two reactions were the same, namely epicatechin, catechin dimer and dehydrogenated catechin dimer. After adding AA, the protective effect of AA on catechin increased first and then decreased with the increase of AA concentration. 0.1 mmolL(exp)-1 AA can inhibit PPO activity in a short time. Within 24 h, 1 mmolL(exp)-1 AA can keep Cat content at 87.88 %. At the concentration of 10 mmolL(exp)-1 AA, excessive AA is oxidized to form a large amount of dehydroascorbic acid (DHAA), which forms an adduct with Cat, promoting the consumption of Cat. The effect of AA on the stability of Cat is time-dependent and dose-dependent.

The mechanism of interaction between lotus rhizome polyphenol oxidase and ascorbic acid: Inhibitory activity, thermodynamics, and conformation analysis.

In this study, the interaction between lotus rhizome polyphenol oxidase (PPO) and ascorbic acid (AA) was discussed from the aspects of inhibitory activity, thermodynamics, and conformation. Results showed that PPO was purified from lotus rhizome by DEAE-52 anion exchange chromatography and Sephadex G-100 gel filtration chromatography, with its optimum substrate being determined as pyrogallic acid. Spectrophotometric and polarographic assays demonstrated that AA exhibited strong inhibitory activity against PPO. Thermodynamics, fluorescence, and circular dichroism spectral analysis showed that hydrophobic interactions caused the formation of AA-PPO complex, leading to the remarkable fluorescence quenching and conformational change of PPO. Atomic force microscopic analysis revealed that binding to AA induced significant changes in the surface morphology and molecular aggregation of PPO molecules. In this study, the interaction mechanism between PPO and AA was proposed for the first time, which provided a theoretical basis for AA to inhibit lotus rhizome browning. PRACTICAL APPLICATIONS: Lotus rhizome, an aquatic vegetable, is prone to enzymatic browning in processing operations, which leads to a decrease in market value and economic loss. At present, ascorbic acid (AA) is widely used in industries as an excellent antioxidant because of its good antibrowning effect and relatively low cost. However, the interaction between the enzymatic browning-related polyphenol oxidase (PPO) from lotus rhizome and ascorbic acid has not been clearly studied. Understanding the mechanism of inhibiting PPO will help to prevent vegetable browning, especially fresh-cut products. The inhibitory effect of AA on PPO in lotus rhizome favors simultaneous use with other types of PPO inhibitors because of their likely synergistic effects.

Sodium bicarbonate reduces the cooked hardness of lotus rhizome via side chain rearrangement and pectin degradation.

In this study, 0.1% (W/V) sodium bicarbonate (SB) solution was used to soften lotus rhizome, and the mechanism was characterized by monoclonal antibodies labeling (mAbs) and atomic force microscopy (AFM). The results showed that the cell wall of lotus rhizome was disintegrated under SB treatment. In addition, the mAbs results revealed that low-esterified homogalacturonan (HG) at the tricellular junction was degraded, the rearrangement of Ara and the interaction between Gal and cellulose may be related to the texture changes. Compared with distilled water treatment, SB treatment reduced the relative content of pectin from 34.1% to 19.1% while increased that of cellulose from 65.9% to 80.9%. AFM results revealed that the height of CSF skeleton decreased from about 32 nm to 1.5 nm. These results clearly demonstrate that cooking with 0.1% SB can soften lotus rhizome through degradation of pectin and arrangement of side chains of rhamnogalacturonan-Ⅰ (RG-Ⅰ).

Effects of air discharge on surface charges and cell walls of Fusarium oxysporum.

Air discharge showed significant inhibition on mycelial growth and spore germination of Fusarium oxysporum, one of the main spoilage fungi in post-harvest lotus roots which is an important economic aquatic vegetable in China. However, the antimicrobial mechanism of air discharge is not clear yet. In the present study, the effects of air discharge on F. oxysporum separated from post-harvest rotten lotus roots were characterized by analyzing surface charges, cell wall permeability, and changes in chitin and chitosan including surface morphology, functional groups, degree of deacetylation, crystallinity, and C/N ratio. After air discharge treatments, alkaline phosphatase leak assay revealed that cell wall permeability of F. oxysporum was magnified. What's more, zeta potentials of F. oxysporum increased and negative charges on cell surfaces decreased. The ordered and compact molecular arrangements of chitin and chitosan in cell walls of F. oxysporum were reduced. The deacetylation degree of chitin and chitosan increased, and the C/N ratios of chitin and chitosan decreased. It was concluded from these results that air discharge caused the transformation in structures of chitin and chitosan, resulting in the exposure of positively charged amino groups and decrease of negative charges on cell surfaces which brought damage to the structure and function of F. oxysporum's cell walls.

Acetic acid reducing the softening of lotus rhizome during heating by regulating the chelate-soluble polysaccharides.

Lotus rhizomes were used to study on the relationship between the cell wall polysaccharides and cooked texture by adding acetic acid. Hardness and scanning electron microscopy results showed that acetic acid treatment can maintain higher hardness and the integrity of the cell wall. Then, the cell walls were sequentially extracted and divided into water-soluble fraction, chelate-soluble fraction (CSF), sodium carbonate-soluble fraction and hemicellulose fraction. The pectin fraction contents, monosaccharides composition, esterification degree and sugar ratios in different groups were evaluated, the results showed that acetic acid increased the total amount of CSF, decreased the esterification degree and less side chain compared that in the solely thermal treatment group. The nanostructures showed that acetic acid treatment maintained longer chain and destroy helical structure of CSF backbone. This work helps us to demonstrate the relationship between polysaccharides structure and cooked texture, and further control the plant-based vegetables processing texture in food industry.

Mechanism of lipid metabolism regulation by soluble dietary fibre from micronized and non-micronized powders of lotus root nodes as revealed by their adsorption and activity inhibition of pancreatic lipase.

Soluble dietary fibre (SDF) of micronized and non-micronized powders of lotus root nodes were investigated based on its adsorption and activity inhibition of pancreatic lipase (PL) by using circular dichroism, fluorescence spectroscopy and modification. Results showed that SDF2 (SDF from micronized powders of lotus root nodes) had stronger PL adsorption and enzyme activity inhibition than SDF1 (SDF from non-micronized powders of lotus root nodes). Specifically, SDF2 showed more binding sites than SDF1 in PL. There were hydrogen bonds and van der Waals interactions between SDF and PL, with Trp on PL probably serving as the main binding site. Carboxyl groups exhibited a stronger inhibition on PL by carboxymethyl and hydroxypropyl modification. The common mechanisms between SDF1 and SDF2 can be attributed to the combination between Trp and carboxyl groups, while the differences may be generated by the variations in structures or chemical groups induced by micronization.

The ratio of chelate-soluble fraction to alcohol insoluble residue is a major influencing factor on the texture of lotus rhizomes after cooking.

The differences in cell wall polysaccharides are considered as a major influencing factor on the texture of plant-based food after cooking. Here, 18 varieties of lotus rhizomes were collected from different regions of China and subjected to analysis, with the aim to identify the key factors that affect the texture of lotus rhizomes after cooking. The texture (hardness) of fresh samples and the samples after thermal treatment for different time periods was examined. The cell wall polysaccharides present in alcohol insoluble residue (AIR) were further subdivided into different fractions, and the composition of monosaccharides in each fraction was analyzed by gas chromatography. We then calculated the sugar ratios to examine the discrepancies in molecular structure among the fractions. Principal component analysis and regression analysis showed that the ratio of chelate-soluble fraction (CSF) to AIR is the major factor affecting the texture of lotus rhizomes after cooking.

Ferulic acid pretreatment alleviates the decrease in hardness of cooked Chinese radish (Raphanus sativus L. var. longipinnatus Bailey).

This study aims to investigate the impact of ferulic acid pretreatment on the hardness of Chinese radish after cooking. Radish slices were immersed in ferulic acid solution and distilled water, respectively. The ferulic acid treated samples showed significantly higher hardness (p < 0.05) than the control samples after cooking, and could keep relatively more integrated cell wall structure after cooking at 100 °C for 30 min. Subsequently, we determined the cell wall fraction contents, sugar ratio and molecular weight distribution of different treated samples. Ferulic acid treated groups showed higher percentage of chelate-soluble fraction (CSF) and sugar ratio 1 than the control groups in both fresh and cooked samples. The CSF had two high molecular weight peaks at 7.7 min and 8.6 min, probably due to the cross-linking with ferulic acid. This research may provide an effective way to decrease the softening of thermally processed fruits and vegetables.

Acetic acid pretreatment improves the hardness of cooked potato slices.

The effects of acetic acid pretreatment on the texture of cooked potato slices were investigated in this work. Potato slices were pretreated with acetic acid immersion (AAI), distilled water immersion (DWI), or no immersion (NI). Subsequently, the cell wall material of the pretreated samples was isolated and fractioned to evaluate changes in the monosaccharide content and molar mass (MM), and the hardness and microscopic structure of the potato slices in different pretreatments before and after cooking were determined. The results showed that the highest firmness was obtained with more intact structure of the cell wall for cooked potato slices with AAI pretreatment. Furthermore, the MM and sugar ratio demonstrated that the AAI pretreated potato slices contained a higher content of the small molecular polysaccharides of cell walls, especially in the hemicellulose fraction. This work may provide a reference for potato processing.

Changes in physicochemical properties related to the texture of lotus rhizomes subjected to heat blanching and calcium immersion.

Pretreatments such as low temperature blanching and/or calcium soaking affect the cooked texture of vegetal food. In the work, lotus rhizomes (Nelumbo nucifera Gaertn.) were pretreated using the following 4 treatments, blanching at 40°C, blanching at 90°C, soaking in 0.5% CaCl2, and blanching at 40°C followed by immersion in 0.5% CaCl2. Subsequently, the cell wall material of pretreated samples was isolated and fractioned to identify changes in the degree of esterification (DE) and monosaccharide content of each section, and the texture of the lotus rhizomes in different pre-treatments was determined after thermal processing with different time. The results showed that the greatest hardness was obtained after blanching at 40°C in CaCl2, possibly attributing to the formation of a pectate calcium network, which maintains the integrity of cell walls. Furthermore, the content of galactose, rhamnose and arabinose decreased due to the breakage of sugar backbones and subsequent damage to cell walls. Our results may provide a reference for lotus rhizome processing.

ATP bioluminescence rapid detection of total viable count in soy sauce.

The adenosine triphosphate (ATP) bioluminescence rapid determination method may be useful for enumerating the total viable count (TVC) in soy sauce, as it has been previously used in food and beverages for sanitation with good precision. However, many factors interfere with the correlation between total aerobic plate counts and ATP bioluminescence. This study investigated these interfering factors, including ingredients of soy sauce and bacteria at different physiological stages. Using the ATP bioluminescence method, TVC was obtained within 4 h, compared to 48 h required for the conventional aerobic plate count (APC) method. Our results also indicated a high correlation coefficient (r = 0.90) between total aerobic plate counts and ATP bioluminescence after filtration and resuscitation with special medium. The limit of quantification of the novel detection method is 100 CFU/mL; there is a good linear correlation between the bioluminescence intensity and TVC in soy sauce in the range 1 × 10(2) -3 × 10(4) CFU/mL and even wider. The method employed a luminescence recorder (Tristar LB-941) and 96-well plates and could analyse 50-100 samples simultaneously at low cost. In this study, we evaluated and eliminated the interfering factors and made the ATP bioluminescence rapid method available for enumerating TVC in soy sauce.

Flow injection chemiluminescence sensor using molecularly imprinted polymers as recognition element for determination of maleic hydrazide.

A novel flow injection chemiluminescence (CL) sensor for the determination of maleic hydrazide (MH) using molecularly imprinted polymer (MIP) as recognition element is reported. The MH-MIP was synthesized by thermal initiated polymerization in methanol, using methacrylic acid (MAA) as functional monomer and ethylene glycol dimethacrylate (EGDMA) as cross-linker in the maleic hydrazide template molecule. Molecular modeling was employed to simulate the possible recognition process of the MIP, and to achieve high selectivity. Then the synthesized MH-MIP was employed as recognition element by packing into flow cell to establish a novel flow injection CL sensor. The CL intensity responded linearly to the concentration of MH in the range 3.5x10(-4)-5.0x10(-2) mg/mL with a detection limit of 6.0x10(-5) mg/mL (3sigma), which is lower than that of conventional methods. The relative standard deviation (RSD) for the determination of 1.0 microg/mL of MH was 2.7% (n=11). The sensor is reusable and has a great improvement in sensitivity and selectivity for CL analysis. As a result, the new MIP-CL sensor had been successfully applied to the determination of MH in vegetable samples.

Determination of catechin in lotus rhizomes by high-performance liquid chromatography.

A novel method was developed to analyze lotus rhizome polyphenolic catechin using high-performance liquid chromatography (HPLC). The retain time of catechin was 14.72 min under the optimized condition. Mass spectrometry was further employed to qualify and quantify the purity of the catechin peak. Good linearity (R=0.9997) was obtained within the range of 50-1,000 ng. The coefficient of variance was determined as 5.2%, with a recovery rate of 97%. The detection and quantification limitations of catechin were 23 ng and 50 ng, respectively. The catechin level was 0.0025% in the lotus rhizome, and 0.011% in the knot of the lotus rhizome (Nelumbo nucifera cv. 'damao jie'). The optimized conditions of HPLC for catechin detection in the lotus rhizome matrix were as follows: the SuperlcosIL™ LC-18 analytical column (150 mm×4.6 mm, 5 µm), methanol-water-acetic acid (10:90:1, volume ratio) as the mobile phase, an UV detector at 280 nm, a flow rate of 0.8 ml/min, column temperature at 30°C, and an injection volume of 10 µl.