HPLC DETERMINATION OF PHENOLIC ACIDS, FLAVONOIDS AND ASCORBIC ACID IN FOUR DIFFERENT SOLVENT EXTRACTS OF ZANTHOXYLUM ACANTHOPODIUM, A WILD EDIBLE PLANT OF MEGHALAYA STATE OF INDIA
Objective: A high-performance liquid chromatographic (HPLC) method using photodiode array (PDA) detector with gradient elution was developed for the quantification of ascorbic acid, free phenolic acids such as gallic acid, methyl gallate, caffeic acid, syringic acid, ferulic acid, para (p)-coumaric acid, sinapic acid and flavonoids (catechin, rutin, quercetin, myricetin, apigenin and kaempferol), in four different solvent extracts like chloroform, methanol, 80% aqueous (aq.) ethanol and 1% aqueous (aq.) acetic acid of wild edible leaves of Zanthoxylum acanthopodium collected from Meghalaya state of India.
Methods: The chromatographic separation was carried out on AcclaimTM120 C 18 column (5 Î¼m particle size, 250 x 4.6 mm), Dionex Ultimate 3000 liquid chromatography and detection were carried out at three different wavelengths (272, 280 and 310 nm) using a mobile phase of acetonitrile and 1% aq. the acetic acid solution with gradient elution.
Results: The experimental results showed a high concentration of ascorbic acid (16.8 mg/g of dry plant material) and gallic acid (0.952 mg/g of dry plant material) in 1% aq. acetic acid extract as compared to the 80% aq. ethanol extract of the plant. The high percentage of recovery (96-103%), low coefficient of variation (R2>0.99) and low limit of detection (LOD) and limit of quantitation (LOQ) confirm the suitability of the method for simultaneous quantification of ascorbic acid and all phenolic compounds in the plant under investigation.
Conclusion: The method can be applied for the simultaneous determination of phenolic acids and flavonoids in different plants and also for the isolation of several bioactive components for the use in pharmaceutical and nutraceutical sector.
Keywords: Phenolic acids, Flavonoids, Ascorbic acid, Different solvent extract, Z. acanthopodium, Gradient HPLC
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