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      Reversed-phase-liquid chromatography method for separation and quantification of gallic acid from hydroalcoholic extracts of Qualea grandiflora and Qualea parviflora

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          Abstract

          Background:

          Qualea parviflora and Qualea grandiflora (Vochysiaceae), commonly known in Brazil as “pau-terra” and “pau-terrinha,” respectively, have been widely used in the treatment of ulcer and gastritis. These therapeutic effects are attributed to various compounds present in the plants, including phenolic compounds such as gallic acid, due to their important antioxidant activity.

          Objective:

          The aim of the present study was to validate a high performance liquid chromatography with diode array detection (HPLC-DAD) method for the quantitative determination of gallic acid in the stem bark of Q. parviflora and Q. grandiflora hydroalcoholic extracts.

          Materials and Methods:

          The chromatography analysis was successfully achieved on a Dionex column, Acclaim ® 120 (250 mm × 4.60 mm, 5 µm) with a gradient elution of water and methanol at a flow rate of 0.8 mL/min and ultraviolet detection at 280 nm.

          Results:

          The validation data, including linearity, precision, specificity, accuracy and robustness of this method demonstrated good reliability and sensitivity.

          Conclusion:

          The method is able to quantify gallic acid in the stem bark of both species. What is more, the chromatographic peaks showed good resolution and there are also the advantages of easy sample preparation and a short time between each injection.

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          Most cited references44

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          Simultaneous determination of catechins, caffeine and gallic acids in green, Oolong, black and pu-erh teas using HPLC with a photodiode array detector.

          A simple and fast HPLC method using a photodiode array detector was developed for simultaneous determination of four major catechins, gallic acid and caffeine. After multiple extractions with aqueous methanol and acidic methanol solutions, tea extract was separated within 20 min using a methanol-acetate-water buffer gradient elution system on a C(18) column. The sample extraction data demonstrated that the single extraction used in the previous studies with aqueous acetonitrile or methanol is not sufficient; the multiple extraction procedure is essential for the quantitative analysis of catechins, phenolic acids and caffeine in teas. Several green, Oolong, black and pu-erh teas were successfully analyzed by this method. The analytical results obtained indicated that green teas contain higher content of catechins [(-)-epigallocatechin gallate, (-)-epigallocatechin, (-)-epicatechin gallate, and (-)-epicatechin] than both Oolong, pu-erh and black teas because fermentation process during the tea manufacturing reduced the levels of catechins significantly. The fermentation process also remarkably elevated the levels of gallic acid in full-fermented pu-erh and black teas. Another interesting finding is the low level of caffeine in Oolong teas, especially in Fujian Oolong tea.
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            Anti-metastasis effects of gallic acid on gastric cancer cells involves inhibition of NF-kappaB activity and downregulation of PI3K/AKT/small GTPase signals.

            Polyphenols are natural antioxidants that are thought to contribute to prevention of cardiovascular disease and malignancy. Although many studies have been carried out to investigate the chemopreventive role of flavonoids, less attention has been focused on phenolic acids. In this study, the aim was to investigate the effect of phenolic acids found abundantly in vegetables, i.e. gallic acid (GA), caffeic acid (CA) and protocatechuic acid (PCA), on the inhibition of gastric adenocarcinoma (AGS) cell metastasis. The results showed 0.01 mM GA induced the same level of cell toxicity as 4.0mM PCA. Using wound-healing assay and Boyden chamber assay, GA had potent inhibitory effects on AGS cell migration. The expression of MMP-2/9 of AGS cells was inhibited by 2.0 microM of GA. It is possible that the suppressive effect of GA on MMP-2/9 might involve the inhibition of NF-kappaB activity. Multiple proteins involved in metastasis and the cytoskeletal reorganization signal pathway, including Ras, Cdc42, Rac1, RhoA, RhoB, PI3K and p38MAPK, were also inhibited by GA. Furthermore, immunoreactivity assay of cytoskeletal F-actin demonstrated a significant inhibitory effect of GA treatment. In conclusion, GA may have the potential to be an effective agent for prevention and treatment of gastric cancer metastasis. Copyright (c) 2010 Elsevier Ltd. All rights reserved.
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              Separation and determination of flavonoids and other phenolic compounds in cranberry juice by high-performance liquid chromatography.

              A HPLC method was developed for the separation and determination of flavonoid and phenolic antioxidants in cranberry juices. Free flavonoid and phenolic compounds were fractionated into neutral and acidic groups by means of a solid-phase extraction method, followed by subsequent HPLC separations. Combined flavonoids and phenolics were hydrolyzed by acid before HPLC analysis. This developed method provides a fast and high resolution of individual flavonoid and phenolic compounds. In cranberry fruit, flavonoids and phenolic acids exist predominantly in combined forms, such as glycosides and esters. A total of 400 mg of total flavonoids and phenolic compounds/l of sample was found in a freshly squeezed cranberry juice, which was distributed as about 44% of phenolic acids and 56% of flavonoids. Benzoic acid was the major phenolic compound. Major flavonoids in the freshly squeezed cranberry juice were quercetin and myricetin.
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                Author and article information

                Journal
                Pharmacogn Mag
                Pharmacogn Mag
                PM
                Pharmacognosy Magazine
                Medknow Publications & Media Pvt Ltd (India )
                0973-1296
                0976-4062
                October 2015
                : 11
                : Suppl 2
                : S316-S321
                Affiliations
                [1 ] Laboratory of Pharmacognosy, University of Brasília – UnB, Brasília-DF, Brazil
                [2 ] Post Graduate Programme in Therapeutic Innovation, Federal University of Pernambuco – UFPE, Recife-PE, Brazil
                [3 ] Pharmacognosy Laboratory, Federal University of Pernambuco – UFPE, Recife-PE, Brazil
                [4 ] Plant Anatomy Laboratory, University of Brasília – UnB; Brasília-DF, Brazil
                Author notes
                Address for correspondence: Dr. Luiz A. L. Soares, Pharmacognosy Laboratory, Department of Pharmaceutical Sciences, Federal University of Pernambuco – UFPE. Prof. Arthur de Sá, s/n, Cidade Universitária, 50740-521 Recife-PE, Brazil. E-mail: lals@ 123456pq.cnpq.br
                Article
                PM-11-316
                10.4103/0973-1296.166062
                4653343
                b925c4f8-3b1f-404f-8e32-cff91c70d18f
                Copyright: © Pharmacognosy Magazine

                This is an open access article distributed under the terms of the Creative Commons Attribution NonCommercial ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non commercially, as long as the author is credited and the new creations are licensed under the identical terms.

                History
                : 26 June 2013
                : 17 August 2013
                : 24 September 2015
                Categories
                Original Article

                Pharmacology & Pharmaceutical medicine
                gallic acid,high-performance liquid chromatography with diode array detection,qualea grandiflora,qualea parviflora,validation

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