Caloscypha fulgens

Polyphenols in tea and orange peel [36 (link)]: Each sample powder (0.2 g) was mixed with 10 mL of 70% methanol, followed by extraction in a water bath at 70 °C, and determining the content of polyphenols using the Folin phenol colorimetric method.
Free amino acids and soluble sugars in tea [37 (link)]: Each sample (0.5 g) was added with 50 mL of boiling water for extraction in a boiling water bath, and, after filtering the tea soup, the ninhydrin colorimetry and the anthrone-sulfuric acid colorimetry were used to determine the content of free amino acids and soluble sugars, respectively.
Determination of orange peel polysaccharide [38 (link)]: For each sample (0.2 g), 40 mL of 80% ethanol was added and extracted by reflux in a 95 °C water bath, followed by adding 100 mL of distilled water for extraction in a boiling water bath, then filtering the tea soup and determining the orange peel polysaccharide using the anthrone sulfuric acid method.
Determination of theaflavins, thearubigins, and theabrownins in tea [35 (link)]: For each sample powder (3 g), 125 mL of boiling water was added, followed by extraction in a boiling water bath, then suction, filtering the tea soup, and determining the content of theaflavins, thearubigins, and theabrownins using the system detection method.
Flavonoids in orange peel [39 (link)]: Each sample (0.2 g) was exposed to ultrasonic extraction with absolute ethanol for 30 min, followed by filtration and determining the content of flavonoids using the aluminum nitrate method.
Soluble protein in orange peel [40 (link)]: Each sample (0.2 g) was extracted with boiling water at 100 °C for 10 min, followed by centrifugation at 3000 rpm for 10 min to collect the supernatant for determination of soluble protein using the Coomassie brilliant blue method.
Determination of hesperidin, synephrine, and limonin in orange peel: Simultaneous determination by HPLC [41 (link)]. Extraction preparation: For each ground orange peel powder (0.1 g), 10 mL of methanol was added, followed by ultrasonic extraction for 30 min, filtration, dilution to 10 mL, and passing 1 mL diluted solution through a 0.22 μm filter membrane for HPLC determination of hesperidin, synephrine, and limonin as described below.
HPLC conditions: an Agilent ZORBAX SB-C 18 chromatographic column (250 mm × 4.6 mm × 5 μm, Agilent, Santa Clara, CA, USA) was used: flow rate, 1 mL·min-1; column temperature, 35 °C; injection volume, 5 μL; detection wavelength, 210 nm and 283 nm; mobile phase A, aqueous phosphoric acid solution with pH = 3.7; mobile phase B, methanol: acetonitrile = 1:1, with the gradient elution as 0~5 min, A:B = 100:0; 5~10 min, A:B = 95:5; 10~20 min, A:B = 75:25; 20~25 min, A:B = 50:50; 25~30 min, A:B = 25:75; 30~40 min, A:B = 5:95; 5~10 min, A:B = 95:5.

A known weight of olives cake and orange peel powders (50 g) was extracted in distilled water at 70 °C for 30 min followed by cooling at room temperature and filtration to remove impurities. The filtrate was dried under vacuum at 50 °C using a rotary evaporator and the extract was saved in sterilized bags and stored at −20 °C before use for physico-chemical analysis. The extraction process was repeated for the filtered residue and substantial quantities of both olive cakes extract (OCE) and orange peel extract (OPE) were obtained. The extraction process was carried out at a low temperature to preserve the phytochemical compounds in the extract [12 (link),13 (link)].

Orange peels were cleaned with DW and dried for 20 h at 50 °C. The dried peels were pulverized in a blender before being activated with ZnCl₂ in a 1:2 (W/W) ratio at 105 °C for 24 h. After that, it was held at 700 °C for 1 h with a nitrogen flow of 50 mL/min in a tubular furnace (T.F. Nabertherm B180 (RT 50/250/13)). It created OPAC in powder form. After being cooled to ambient temperature, the activated carbon was refluxed with 1N HCl for 2 h to eliminate the alkali and then washed with DW to achieve a neutral pH. It was then dried for four hours at 105 °C.

In this study, Experimental research and field studies on plant material (Orange peels), including the collection of plant waste material, complies with relevant institutional, national, and international guidelines and legislation.

Orange peels were gathered from a local market in Alexandria, Egypt, cleaned with distilled water (DW), and dried at 50 °C for 24 h. The dried peels were crushed in a mixer and put away until they were needed. Stock solution of Cr⁶⁺ ions was organized by dissolving 2.83 g of K₂Cr₂O₇ in 1 L DW. Potassium dichromate (K₂Cr₂O₇, M.W 294.185 g, assay 99.5%) and Ferric nitrate anhydrous (Fe(NO₃)₃, M.W. 241.86 g, assay 98%) were gotten from ADWIC, El-Nasr Chemical Company, Egypt. BDH Chemicals LTD provided the 1,5-diphenylcarbazide used as a substance for Cr⁶⁺ ions, while Universal Fine Chemicals PVT-LTD in Mumbai, India provided ZnCl₂ (M.W.136.30 g, assay 99.5%). From SD Fine-Chem. Limited (SD FCL), we got HCl (M.W. 36.46 g, test 30–34%). For the synthesis of magnetite Fe₃O₄, ferrous sulphate (FeSO₄^.7H₂O, M.W. 278.01 g, assay 98.5%) was acquired from Alpha Chemika in India. None of the compounds was further purified before usage.