The ultraviolet analysis (UV/VIS) was carried out employing a Shimadzu UV-visible recording spectrophotometer model-UV 240 (NRC, Cairo, Egypt). The nuclear magnetic resonance (1H- and 13C-NMR) analysis was performed via Varian Unity Inova (IET, Urbana, IL, USA). Mass spectroscopy was performed via Finnigan MAT SSQ 7000, 70 ev (Adaptas Solutions, Palmer, MA, USA). Flash column chromatography was employed using a Sephadex LH-20 (Sigma-Aldrich, Taufkirchen, Germany), obtained from Pharmacia Fine Chemicals, and silica gel (0.063–0.200 mm). Paper Chromatography (PC) was performed on Whatman No.1 sheets (Whatman Led. Maid Stone, Kent, UK) to quantify flavonoids and sugars in the extract. Thin-layer chromatography (TLC) analysis was performed on F254 plates. The solvent mixtures for separation were applied (n-butanol: acetic acid: water 4:1:5 upper phase, 15% acetic acid: water: glacial acetic acid: 85:15).
Whatman no 1 sheets
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Whatman No. 1 sheets are a type of filter paper designed for general laboratory filtration applications. They are made of high-quality cellulose and provide consistent, reliable filtration performance.
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3 protocols using whatman no 1 sheets
1
Spectroscopic Analysis of Organic Compounds
2
Flavonoid Aglycone Extraction and Analysis
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Flavonoid glycosides 5, 6, 7, and 8 (5 mg) were heated in a solution of 10% HCl. After 5 h, the mixture was extracted with ethyl acetate to afford crude aglycones. The crude mixture was separated and analyzed by TLC utilizing reference standards. The aqueous layer was analyzed by paper chromatography Whatman No. 1 sheets in the presence of reference standards employing a solvent system (n-BuOH-AcOH-H2O 4:1:5 upper layer) to provide the sugar content of the flavonoid [22 (link)].
3
Phytochemical Characterization of Compounds
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The NMR spectra were run at 300 and 500 (1H), 75 and 125 (13C) MHz, on Varian Mercury 300 and JEOL GX-500 NMR spectrometers, respectively. The chemical shifts (δ) are reported in ppm downfield to TMS in the appropriate deuterated solvent. For ESI-MS analyses, LCQ (Finnigan MAT 95, Bremen, Germany) and LTQ-FT-MS spectrometers were used, while HR-ESI-MS (Thermo Electron, Finnigan, Germany) was used for HR-ESI-MS analyses. UV spectrophotometer (JASCO V-630) was used for analysis of pure samples in MeOH and in different UV shift reagents.
Material for column chromatography (CC), including microcrystalline cellulose, polyamide S 6 and Sephadex LH-20, as well as Whatman No. 1 sheets used for paper chromatography were purchased from sources described in our previous literature.[11 (link)] Isolated compounds were detected using Naturstoff[11 (link)] and/or FeCl3 (1% in ethanol) spray reagents. Solvent systems S1 (n-BuOH/HOAc/H2O; 4:1:5 v/v/v top layer), S2 (HOAc/H2O; 15:85 v/v) and S3 (n-BuOH/iso-propanol/H2O; 4:1:5, v/v/v top layer) were used. Indomethacin was obtained from Epico, Egypt; paracetamol and silymarin from Sedico, 6th October, Egypt; carrageenan from Sigma, USA; alanine aminotransferase (ALT), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) kits were purchased from Marcy’ Étoile (France).
Material for column chromatography (CC), including microcrystalline cellulose, polyamide S 6 and Sephadex LH-20, as well as Whatman No. 1 sheets used for paper chromatography were purchased from sources described in our previous literature.[11 (link)] Isolated compounds were detected using Naturstoff[11 (link)] and/or FeCl3 (1% in ethanol) spray reagents. Solvent systems S1 (n-BuOH/HOAc/H2O; 4:1:5 v/v/v top layer), S2 (HOAc/H2O; 15:85 v/v) and S3 (n-BuOH/iso-propanol/H2O; 4:1:5, v/v/v top layer) were used. Indomethacin was obtained from Epico, Egypt; paracetamol and silymarin from Sedico, 6th October, Egypt; carrageenan from Sigma, USA; alanine aminotransferase (ALT), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) kits were purchased from Marcy’ Étoile (France).
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