In situ detection and identification of hesperidin crystals in satsuma mandarin (Citrus unshiu) peel cells.

INTRODUCTION: Hesperidin, a flavonoid known to have important pharmacological effects, accumulates particularly in the peels of satsuma mandarin (Citrus unshiu). Although histochemical studies have suggested that hesperidin forms crystals in some tissues of the Rutaceae and Umbelliferae, there has been no rigorous in situ detection or identification of hesperidin crystals in C. unshiu. OBJECTIVE: To characterise the chemical component of the crystals found in C. unshiu peels using Raman microscopy. METHODS: Sections of C. unshiu peels were made. The distribution and morphology of crystals in the sections were analysed microscopically. Raman microscopy was used to detect hesperidin in the sections directly. RESULTS: The crystals were more abundant in immature peel and were observed particularly in areas surrounding vascular bundles, around the border between the flavedo and albedo layers and just below the epidermal cells. In the morphological analysis by scanning electron microscopy, needle-shaped crystals aggregated and formed clusters of spherical crystals. Spectra obtained by Raman microscopy of the crystals in the peel sections were consistent with those of the hesperidin standard. CONCLUSION: This study showed the detailed distribution of crystals in C. unshiu peels and their main component was identified using Raman microscopy to be hesperidin for the first time.

Total fractionation of green tea residue by microwave-assisted alkaline pretreatment and enzymatic hydrolysis.

Total refinery of constituents of green tea residue was achieved by combination of microwave-assisted alkaline pretreatment and enzymatic hydrolysis. Alkaline pretreatment was effective at separating pectic polysaccharides, protein, phenolic compounds and aliphatic compounds (probably originating from cuticular components), and the solubilization rate was attained 64­74% by heating at 120­200 °C. The higher heating value (HHV) of alkali-soluble fraction attained 20.1 MJ/kg, indicating its usability as black-liquor-like biofuel. Successive cellulolytic enzymatic hydrolysis mainly converted cellulose into glucose and attained the maximum solubilization rate of 89%. Final residue was predominantly composed of aliphatic cuticular components with high proportion in 9,10,18-trihydroxyoctadecanoic acid (30.1­48.6%). These cuticular components are potential alternative feedstock for aliphatic compounds commonly found in oil plants.

Compositional analysis of leaf cuticular membranes isolated from tea plants (Camellia sinensis L.).

Chemical constituents of cuticular membranes (CMs) isolated from three tea cultivars (Camellia sinensis (L.) O. Kuntze cvs. Yabukita, Samidori and Gokou) were compared. All CMs from the adaxial side of the leaves showed higher accumulation of wax, cutin and polysaccharide, while those from the abaxial side were abundant in cutan, showing the adaptation of the adaxial side to abiotic stresses, such as wind and rain, in contrast to the abaxial side, which provides defence against pathogens. Yabukita, a major tea cultivar in Japan, developed thick CMs while Samidori and Gokou, shade-cultivated tea cultivars, had lighter CMs, reflecting their thin and soft leaves. CMs rapidly accumulated 9,10-epoxy-18-hydroxyoctadecanoic acid at a very early stage of leaf development. Additionally, shade treatment did not influence cutin biosynthesis in CMs, reflecting high adaptation of tea leaves under low light levels.

A novel saccharification method of starch using microwave irradiation with addition of activated carbon.

Activated carbons were investigated for their heat catalytic effects to improve saccharification of starch by autohydrolysis in water under microwave electromagnetic field, and the results were compared with graphite and carbon nanotubes. The activated carbons with low adsorptive capacity of maltose showed high saccharification rate, while those with high adsorptive capacity exhibited low saccharification. In addition, the former activated carbons decreased the saccharification temperature by 10-30°C. Maltooligosaccharides produced in the presence of the latter activated carbons were recovered by desorption with 50% aqueous ethanol. The results indicated that both adsorptive capacities of maltooligosaccharides and catalytic effects of hot spots arisen from the uneven surface structure of activated carbons might contribute to the improvement in starch saccharification.

Microwave heating of tea residue yields polysaccharides, polyphenols, and plant biopolyester.

Microwave heating was used to produce aqueous-soluble components from green, oolong, and black tea residues. Heating at 200-230 degrees C for 2 min extracted 40-50% of polysaccharides and 60-70% of the polyphenols. Solubilization of arabinose and galactose by autohydrolysis occurred with heating above 170 degrees C, whereas heating above 200 degrees C was necessary to solubilize xylose. Catechins were soluble in water by heating at low temperature (110 degrees C); however, new polyphenols having strong antioxidant activity were produced above 200 degrees C. The amount of solubilized materials and antioxidant activity increased with increased fermentation of harvested tea leaves (green tea < oolong tea < black tea). Cutin, a plant biopolyester, remained in the residue after heating as did cellulose and lignin/tannin. The predominant cutin monomer that was recovered was 9,10-epoxy-18-hydroxyoctadecanoic acid, followed by dihydroxyhexadecanoic acid and 9,10,18-trihydroxyoctadecanoic acid.