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3 protocols using analytical grade ethanol

1

Nelumbo nucifera Embryo Extraction

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A Plumula Nelumbinis sample was purchased from Beijing Tong-Ren-Tang
Group Co., Ltd. (Beijing, China), and identified by Dr. Ailiang Chen (Institute
of Quality Standards and Testing Technology for Agro-products, Chinese Academy
of Agricultural Sciences, Beijing, China) as the embryo of the seed of
Nelumbo nucifera Gaertn. Analytical grade ethanol,
HPLC-grade acetonitrile and water were purchased from Thermo Fischer Scientific
(Fremont, CA, USA); ammonium acetate, formic acid and other reagents were
purchased from Sigma (St. Louis, MO, USA).
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2

Quantification of Volatile Phenols in Wines

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Analytical grade volatile phenols (guaiacol, 4-methylguaiacol, 4-ethylguaiacol, 4-ethylphenol, o-cresol, m-cresol, p-cresol, and eugenol) and deuterated NMR solvents (d6-ethanol, D2O, and DCl) were purchased from Sigma-Aldrich (Castle Hill, NSW, Australia). Deuterium-labelled internal standards (d3-guaiacol, d3-4-methylguaiacol, and d4-4-ethylphenol) were sourced from CDN Isotopes (Pointe-Claire, Quebec, Canada). Analytical grade ethanol, tartaric acid, and sodium hydroxide were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Food grade (>98% purity) α-, β-, and γ-CDs were supplied by IMCD Group (Adelaide, SA, Australia). Model wine was prepared by dissolving tartaric acid (5 g/L) in aqueous ethanol (12% alcohol by volume) and adjusting the pH to 3.5 by dropwise addition of 1 M sodium hydroxide. Stock solutions of internal standards and volatile phenols were prepared volumetrically in absolute ethanol and stored at −20 °C, with working solutions prepared in model wine and stored at 4 °C.
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3

Comprehensive Phytochemical Analysis Protocol

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Analytical grade ethanol, methanol, hydrochloric acid, sodium acetate, and glacial acetic acid were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). H2SO4 (98%) was procured from RCI Labscan (Rongmuang, Thailand). Na2CO3 was procured from Chem-Supply Pty Ltd. (Adelaide, Australia). Folin-2,2'azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), Ciocalteu reagent, 2,4,6-tripyridyl-s-triazine (TPTZ), 2,2-diphenyl-1-picrylhydrazyl (DPPH), L-ascorbic acid, quercetin, vanillin, gallic acid, AlCl3.6H2O, FeCl2, Ferrous sulphate, hydrogen peroxide (H2O2), 3-hydroxybenzoic acid, catechin, ferrozine, trichloroacetic acid, potassium ferricyanide, and FeCl3 were acquired from Sigma-Aldrich (Castle Hill, New South Wales, Australia). 2-hydroxybenzoic acid, catechin, coumaric acid, gallic acid, sinapic acid, syringic acid, quercetin-3-rhamnoside, p-hydroxybenzoic acid, epicatechin gallate and phloretic acid were used as standards in HPLC-PDA analysis.
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