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2 protocols using anhydrous aluminium chloride

1

Phytochemical Extraction and Analysis

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Methanol (I0962907) was purchased from Merck KGaA Germany. Pyrogallol solution was purchased from Oxford Labs (India). Other chemicals such as Carboxymethyl cellulose, sulfanilamide, Ethylenediamine tetraacetic acid (EDTA), Griess reagent, N-1- naphthylethyleneamine dihydrochloride, thiobarbituric acid, anhydrous aluminium chloride, copper sulfate, sodium phosphate dibasic heptahydrate and sodium phosphate monobasic monohydrate, phosphoric acid, picric acid, sodium carbonate, sodium hydroxide, sodium-potassium tartrate, gallic acid, Folin-Ciocalteu reagent, and quercetin were purchased from Sigma Aldrich United States.
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2

Quantification of Polyphenols and Flavonoids

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Estimation of total polyphenols and flavonoids was performed using UV-visible spectrophotometer (Sagar Scientific Works, Ambala cantt, India), Nicolet evolution 100 (Thermo) was used for measuring the absorbance at 765 nm (for polyphenols) and at 415 nm (for flavonoids) using the following authentic and reagents:
Gallic acid was acquired from E-Merck, Darmstadt, Germany. • Quercetin was acquired from Sigma-Aldrich, Hamburg, Germany. • Sodium carbonate solution (74.2 g) was dissolved in 1000 mL distilled water (0.7 M solution) (Makky et al., 2012) . • Folin-Ciocalteu reagent was purchased from Sd fine Chem.
Limited, Mumbai, India. • Anhydrous aluminium chloride (2 g; Sigma-Aldrich, Hamburg, Germany) was cautiously dissolved in 100 mL of spectroscopic methanol (Aliyu et al., 2009) .
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