The thin layer chromatography (TLC) method was employed to separate the plant extract and evaluate their constituents. Ten microliter of each plant extract (1 mg/mL) was spotted on TLC plates (Whatman Inc., Florham Park, U.S.A) and were placed in TLC chambers containing different mobile phases with different polarities, namely EMW (ethyl acetate (Sigma Aldrich, Darmstadt, Germany)/methanol/water (40:5.4:4), CEF (chloroform (Merck, Darmstadt, Germany)/ethyl acetate (Sigma Aldrich, Darmstadt, Germany)/formic acid (Sigma Aldrich, Darmstadt, Germany) (5:4:1, v/v/v), and BEA (benzene (Sigma Aldrich, Darmstadt, Germany)/ethanol (Sigma Aldrich, Darmstadt, Germany)/ammonium hydroxide (Merck, Darmstadt, Germany) (90:10:1, v/v/v). The plates were developed and thereafter, visualized under UV light at wavelengths of 254 nm and 365 nm. The plates were sprayed with p-anisaldehyde-sulphuric acid (Sigma Aldrich, Darmstadt, Germany) and heated at 100 °C to allow color development to determine the phytochemical fingerprints of the extracts.
P anisaldehyde sulphuric acid
Manufactured by Merck Group
Sourced in Germany
P-anisaldehyde-sulphuric acid is a chemical reagent used in analytical chemistry and organic synthesis. It is a solution of p-anisaldehyde in concentrated sulfuric acid. The product is commonly used as a staining agent in thin-layer chromatography (TLC) to visualize organic compounds.
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2 protocols using p anisaldehyde sulphuric acid
1
Phytochemical Fingerprinting of Plant Extracts
2
Isolation of Bornyl Acetate from Artemisia tridentata
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B. tridentata essential oil (0.856 g, composite sample of female and male essential oils) was separated by column chromatography (CC) over 30.09 g of Si-Gel (230-400 mesh; Merck, Darmstadt, Germany) as the stationary phase. All the solvents employed for CC, and in the next sections were purchased from Cicarelli (San Lorenzo, Santa Fe, Argentina) and distilled prior to use. The CC elution was performed isocratically with a mobile phase mixture of petroleum ether-AcOEt (20:1) at a flow rate of 2.4 mL/min. To monitor CC separation, TLC was employed (Si-Gel60F 254 ; Merck) using the same mobile phase and p-anisaldehyde-sulphuric acid (Sigma-Aldrich) as a spray reagent. The chromatographic separation yielded 30 mg of pure bornyl acetate.
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