Ellagic acid from pomegranate peel: Consecutive countercurrent chromatographic separation and antioxidant effect
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Abstract:
Ellagic acid is one of the most representative natural antioxidants, and is rich in pomegranate peel. In this study, a consecutive countercurrent chromatographic (CCC) separation method was established to improve the preparative efficiency of ellagic acid from pomegranate peel. By optimizing the solvent system, sample size and flow rate, 280 mg of ellagic acid was obtained from 5 g of crude sample from pomegranate peel by CCC after six consecutive injections. Moreover, the values of EC50 for ellagic acid in scavenging ABTS·+ and DPPH· were 4.59 ± 0.07 and 10.54 ± 0.07 μg/ml, respectively, indicating a strong antioxidant activity. This study not only established a high-throughput method for the preparation of ellagic acid, but also provided a successful example for the development of and research on other natural antioxidants.Keywords:
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TLC densitometry method for quantification of gallic acid and ellagic acid using HPTLC is developed. HPTLC is suitable method for estimation of chemical constituent present in plant material. This is the first report of quantification of these two biologically active compounds as gallic acid and ellagic acid using HPTLC from this plant. Quantification of gallic acid and ellagic acid was carried out from the methanol extract by using solvent system of Toluene: Ethyl acetate: Formic acid (6:4:0.8 v/v/v). The Rf value of Gallic acid and ellagic acid are 0.43 and 0.38 respectively. The linearity ranges of gallic acid (100 to 600 ng) and ellagic acid (100 to 600 ng) with correlation coefficient (r- values) of 0.9994 and 0.9997 respectively. The amount of gallic acid and ellagic acid was 3.01 and 11.09 µg/ml in leaf and 4.97 and 18.30 µg/ml in stem respectively. Quantification of gallic acid and ellagic acid showed good separation and resolution from other constituent of extract. The main advantage of this method is simplicity, accuracy and selectivity. Its main advantages are its simplicity, accuracy and selectivity. This method can also be used for estimation of these compounds in other herbal preparation and may be useful for standardization purposes.
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The method for determining the content of gallic acid and ellagic acid in Granati Pericarpium was established by HPLC. Using the method, the content of raw and charred Granati Pericarpium was determined. By comparison, it was found that the content of gallic acid and ellagic acid increased first and then reduced during processing. When processed on an appropriate degree, the content reached the maximum. The result indicated that gallic acid and ellagic acid can be used as indicators to control the processing degree of charred Granati Pericarpium.
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TLC densitometry method for quantification of gallic acid and ellagic acid using HPTLC is developed. This is the first report of quantification of these two biologically active compounds as gallic acid and ellagic acid using HPTLC from this plant. Quantification of gallic acid and ellagic acid was carried out from the methanol extract by using solvent system of Toluene: Ethyl acetate: Formic acid (6:4:0.8 v/v/v). The Rf value of Gallic acid and ellagic acid are 0.43 and 0.38 respectively. The linearity ranges of gallic acid (100 to 600 ng) and ellagic acid (100 to 600 ng) with correlation coefficient (r- values) of 0.9994 and 0.9997 respectively. The amount of gallic acid and ellagic acid was 3.01 and 11.09 μg/ml in leaf and 4.97 and 18.30 μg/ml in stem respectively. Quantification of gallic acid and ellagic acid showed good separation and resolution from other constituent of extract. Its main advantages are its simplicity, accuracy and selectivity. This method can also be used for estimation of these compounds in other herbal preparation and may be useful for standardization purposes.
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Objective To study the pharmacokinetic character of ellagic acid and gallic acid of pomegranate peel extract in rats. Methods After the rats were administered by pomegranate peel extract through ig, HPLC was used to determine the concentration of ellagic acid and gallic acid in blood plasma at different times, pharmacokinetic software Win Nonlin 5.2 was used to process concentration-time date. Results Ellagic acid and gallic acid were shown to be two room distribution model in rats, the main pharmacokinetic parameters of ellagic acid and gallic acid: T1/2(ka)=0.150, 0.779 h,T1/2α=0.570, 0.856 h, T1/2β=6.73, 5.59 h, Tmax=0.50, 1.38 h, Cmax=7.29, 7.15 μg/m L, α=1.21, 0.81 h-1, β=0.103, 0.124 h-1,Ka= 0.462, 0.889 h-1. Conclusion It indicates that the pharmacokinetic profile of ellagic acid and gallic acid are rapid absorption and distribution after oral administration pomegranate peel extraction, peak time is short, the absorption of gallic acid is greater than ellagic acid in pomegranate peel.
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Ashwagandharishta is a polyherbal hydro-alcoholic formulation and is used as immunomodulator to promote the health and longevity by increasing defence against disease and also known for its usefulness in the treatment of hypercholesterolemia. A simple, precise, robust and accurate HPTLC method has been established for the determination of gallic acid and ellagic acid in Ashwagandharishta–T and Ashwagandharishta-M prepared by traditional and modern methods respectively and also in its marketed formulation. The developed HPTLC method was validated in terms of precision, accuracy, LOD, LOQ and specificity. The amount of gallic acid in Ashwagandharishta-T, M and its marketed formulation was found to be 0.0281, 0.0279 and 0.0280% w/w respectively while ellagic acid was found to be 0.0191, 0.0189 and 0.0188% w/w respectively. This is the first report for quantification of gallic acid and ellagic acid in Ashwagandharishta by HPTLC. Furthermore, no TLC densitometric methods have been reported for the quantification of gallic acid and ellagic acid from Ashwagandharishta.
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Gallic acid and ellagic acid are two widely occurring phenolic compounds of plant origin, to which many biological activities including anticancer and antiviral activity have been attributed. A simple HPTLC method has been developed for the simultaneous quantification of gallic acid and ellagic acid. The method was validated for precision, repeatability, and accuracy. Instrumental precision was found to be 0.083 and 0.78, and the repeatability of the method was found to be 1.07 and 1.50 (% CV) for gallic acid and ellagic acid, respectively. The accuracy of the method was checked by a recovery study conducted at two different levels and the average percentage recovery was found to be 101.02% for gallic acid and 102.42% for ellagic acid. The above method was used for the quantification of gallic acid and ellagic acid content in seeds of Abrus precatorius Linn., whole plant of Phyllanthus maderaspatensis Linn., and flowers of Nymphaea alba Linn.The proposed HPTLC method for the simultaneous quantification of gallic acid and ellagic acid was found to be simple, precise, specific, sensitive, and accurate and can be used for routine quality control of herbal raw materials and for the quantification of these compounds in plant materials.
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Background: Solanum dubium is a plant believed to have several therapeutic effects including anti-asthmatic properties. The objective of this study was to investigate the quantitative estimation of Gallic acid and Ellagic acid from the seed extract of Sudanese Solanum dubium. Methods: A simple and rapid high-performance thin-layer chromatographic method was developed and validated for quantitative estimation of Gallic acid and Ellagic acid from the seed extract of Sudanese Solanum dubium. Results: Ellagic acid and Gallic acid were quantified by using HPTLC. The seeds were found to contain 1.1% w/w of Ellagic acid and 2.1% w/w of Gallic acid in extract. Gallic acid and Ellagic acid were chromatographed on silica gel 60 F254 TLC plate using Toluene: Ethyl acetate – Methanol – Formic acid (3:3:1:0.4 v/v/v/v) as mobile phase and quantified by densitometric scanning at 280 nm. The method was found to give compact spots for Gallic acid (Rf 0.35) and Ellagic acid (Rf 0.21). The linear regression analysis data for standard Gallic and Ellagic acids spots showed good linear relationship with r2 = 0.988 and 0.994 respectively, in the concentration range 100-3000 ng/spot, accurate (99.23-102.7%), (98.7-100.2%); precise (% RSD ≤ 2); robust (% RSD ≤ 2) and specific. The LOD and LOQ of the method were found as 653 and 396.8ng/spot and 215.5and 130 ng/spot, of Gallic acid and Ellagic acid respectively. Conclusion: The method was validated for precision, recovery and repeatability as per the International Conference on Harmonization Guidelines for Gallic acid and Ellagic acid. Statistical analysis of the data showed that the method is precise, accurate, reproducible and selective for the analysis of Gallic acid and Ellagic acid.
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Compared with abundant catechins, strictinin is a minor constituent in teas and has been demonstrated to possess inhibitory potency on influenza virus. In this study, strictinin was found as the major phenolic compound in Pu'er teas produced from leaves and buds of wild trees. Due to its thermal instability, strictinin, in tea infusion or in an isolated form, was completely decomposed to ellagic acid and gallic acid after being autoclaved for 7 minutes. A plaque reduction assay was employed to compare the relative inhibitory potency between strictinin and its thermally degraded products against human influenza virus A/Puerto Rico/8/34. The results showed that the antiviral activity of ellagic acid regardless of the presence or absence of gallic acid was significantly higher than that of strictinin. Thermal degradation of strictinin to ellagic acid and gallic acid seems to be beneficial for the preparation of Pu'er teas in terms of enhancing antiviral activity.
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