Detection of antioxidant constituents from Persicaria hydropiper using LC-UV-DAD-ESIM/MS.

The antioxidant activity of the crude methanolic extract and fractions of the aerial parts of Persicaria hydropiper or ‘kesum’ were investigated to evaluate their potential application as health supplement. Dried aerial part of the plant samples were extracted with methanol and the methanol crude ex...

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Main Author: Noor Hashim, Noor Haslinda
Format: Thesis
Language:English
Published: 2011
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Online Access:http://psasir.upm.edu.my/id/eprint/27001/1/IB%202011%2018R.pdf
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id my-upm-ir.27001
record_format uketd_dc
institution Universiti Putra Malaysia
collection PSAS Institutional Repository
language English
topic Antioxidants
Persicaria
Liquid chromatography
spellingShingle Antioxidants
Persicaria
Liquid chromatography
Noor Hashim, Noor Haslinda
Detection of antioxidant constituents from Persicaria hydropiper using LC-UV-DAD-ESIM/MS.
description The antioxidant activity of the crude methanolic extract and fractions of the aerial parts of Persicaria hydropiper or ‘kesum’ were investigated to evaluate their potential application as health supplement. Dried aerial part of the plant samples were extracted with methanol and the methanol crude extract was fractionated by liquid-liquid extraction into five different fractions. All fractions were analyzed for total phenolic content utilizing Folin Ciocalteau assay. The antioxidative potential of all fractions were also evaluated using 1,1-diphenyl-2-picryhydrazyl radical (DPPH), ferric thiocyanate (FTC) and xanthine oxidase inhibition (XO) methods. Furthermore, the metabolite profiling of the bioactive antioxidant fractions was accomplished using liquid chromatography coupled with mass spectrometry (LCMS). The use of LCMS techniques is an important approach for online identification of plant constituents. The amount of the total phenolics, varied widely in the fractions and ranged from 0.00 to 224.4 mg GAE/100 g dry extract. The highest level was found in butanol fraction with 224.4 mg GAE/100 g dry extract, followed by ethyl acetate (68.9 mg GAE/100 g dry extract), dichloromethane (8.7 mg GAE/100 g dry extract), aqueous (6.4 mg GAE/100 g extract) and hexane (4.3 mg GAE/100 g dry extract) fractions. The DPPH assay revealed that the ethyl acetate and butanol fractions of P. hydropiper were the most active fractions with IC50 values of 25.5 and 28.6 μg/ml, respectively. Dichloromethane fraction showed moderate activity with IC50 value of 94.2 μg/ml. The lower radical scavenging activity was observed in aqueous and hexane fractions with IC50 values ranged from 110 to 380 μg/ml. In comparing the antioxidant activity using ferric thiocyanate assay, all fractions effectively inhibited higher lipid oxidation compared to α-tocopherol. Except for the hexane fraction, the inhibition of the other fractions were comparable to the known synthetic antioxidant, butylated hydroxytoluene (BHT). Of the fractions assayed for xanthine oxidase inhibitory activity, butanol fraction showed the most active with IC50 value of 32.3 μg/ml followed by ethyl acetate fraction with IC50 value of 166.9 μg/ml. The ethyl acetate and butanol fractions exhibited good antioxidant activity in all the assay tested. The LC-DAD-ESIMS chemical profiling method was developed for the identification of phytochemical constituents in the antioxidative fractions of P. hydropiper. The phytochemical profile of ethyl acetate fraction has identified fourteen compounds together with two unidentified compounds. The compounds were identified as quercetin (1), quercetin-3-O-glucoside (3), galloyl quercetin-3-O-glucoside (5), galloyl quercetin-3-O-rhamnoside (6), quercetin-3-O-rhamnoside (7), kaempferol-3-O-glucoside (13), galloyl kaempferol-3-O-glucoside (14),hydropiperoside (31), vanicoside A (34), vanicoside B (35), vanicoside D (38), rhamnetin (87), apigenin-3-O-glucoside (89) and kaempferol rutinoside (90). From the ethyl acetate fraction, six compounds; 1, 3, 5, 7, 87 and 3,5-dihydroxy-4-methoxybenzoic acid (88) have been isolated and identified using NMR spectroscopy. Phytochemical investigation of the bioactive butanol fraction of P. hydropiper using LC-DAD-ESIMS/MS led to the identification of nine compounds including rhamnazin-3-sulphate (91), quercetin-3-O-glucuronide (8), luteolin-3-O-glucoside (92), kaempferol-3-O-sulphate (93), quercetin-3-O-rhamnoside (7), apigenin-3-O-glucoside (89), quercitrin 7-sulphate (94), luteolin-7-sulphate (95) and isorhamnetin-3-sulphate (persicarin) (96). All compounds were relatively identified based on the MS/MS and UV data in comparison with those of the standard compounds.
format Thesis
qualification_level Master's degree
author Noor Hashim, Noor Haslinda
author_facet Noor Hashim, Noor Haslinda
author_sort Noor Hashim, Noor Haslinda
title Detection of antioxidant constituents from Persicaria hydropiper using LC-UV-DAD-ESIM/MS.
title_short Detection of antioxidant constituents from Persicaria hydropiper using LC-UV-DAD-ESIM/MS.
title_full Detection of antioxidant constituents from Persicaria hydropiper using LC-UV-DAD-ESIM/MS.
title_fullStr Detection of antioxidant constituents from Persicaria hydropiper using LC-UV-DAD-ESIM/MS.
title_full_unstemmed Detection of antioxidant constituents from Persicaria hydropiper using LC-UV-DAD-ESIM/MS.
title_sort detection of antioxidant constituents from persicaria hydropiper using lc-uv-dad-esim/ms.
granting_institution Universiti Putra Malaysia
granting_department Institute of Bioscience
publishDate 2011
url http://psasir.upm.edu.my/id/eprint/27001/1/IB%202011%2018R.pdf
_version_ 1747811569343922176
spelling my-upm-ir.270012015-05-20T07:58:19Z Detection of antioxidant constituents from Persicaria hydropiper using LC-UV-DAD-ESIM/MS. 2011-06 Noor Hashim, Noor Haslinda The antioxidant activity of the crude methanolic extract and fractions of the aerial parts of Persicaria hydropiper or ‘kesum’ were investigated to evaluate their potential application as health supplement. Dried aerial part of the plant samples were extracted with methanol and the methanol crude extract was fractionated by liquid-liquid extraction into five different fractions. All fractions were analyzed for total phenolic content utilizing Folin Ciocalteau assay. The antioxidative potential of all fractions were also evaluated using 1,1-diphenyl-2-picryhydrazyl radical (DPPH), ferric thiocyanate (FTC) and xanthine oxidase inhibition (XO) methods. Furthermore, the metabolite profiling of the bioactive antioxidant fractions was accomplished using liquid chromatography coupled with mass spectrometry (LCMS). The use of LCMS techniques is an important approach for online identification of plant constituents. The amount of the total phenolics, varied widely in the fractions and ranged from 0.00 to 224.4 mg GAE/100 g dry extract. The highest level was found in butanol fraction with 224.4 mg GAE/100 g dry extract, followed by ethyl acetate (68.9 mg GAE/100 g dry extract), dichloromethane (8.7 mg GAE/100 g dry extract), aqueous (6.4 mg GAE/100 g extract) and hexane (4.3 mg GAE/100 g dry extract) fractions. The DPPH assay revealed that the ethyl acetate and butanol fractions of P. hydropiper were the most active fractions with IC50 values of 25.5 and 28.6 μg/ml, respectively. Dichloromethane fraction showed moderate activity with IC50 value of 94.2 μg/ml. The lower radical scavenging activity was observed in aqueous and hexane fractions with IC50 values ranged from 110 to 380 μg/ml. In comparing the antioxidant activity using ferric thiocyanate assay, all fractions effectively inhibited higher lipid oxidation compared to α-tocopherol. Except for the hexane fraction, the inhibition of the other fractions were comparable to the known synthetic antioxidant, butylated hydroxytoluene (BHT). Of the fractions assayed for xanthine oxidase inhibitory activity, butanol fraction showed the most active with IC50 value of 32.3 μg/ml followed by ethyl acetate fraction with IC50 value of 166.9 μg/ml. The ethyl acetate and butanol fractions exhibited good antioxidant activity in all the assay tested. The LC-DAD-ESIMS chemical profiling method was developed for the identification of phytochemical constituents in the antioxidative fractions of P. hydropiper. The phytochemical profile of ethyl acetate fraction has identified fourteen compounds together with two unidentified compounds. The compounds were identified as quercetin (1), quercetin-3-O-glucoside (3), galloyl quercetin-3-O-glucoside (5), galloyl quercetin-3-O-rhamnoside (6), quercetin-3-O-rhamnoside (7), kaempferol-3-O-glucoside (13), galloyl kaempferol-3-O-glucoside (14),hydropiperoside (31), vanicoside A (34), vanicoside B (35), vanicoside D (38), rhamnetin (87), apigenin-3-O-glucoside (89) and kaempferol rutinoside (90). From the ethyl acetate fraction, six compounds; 1, 3, 5, 7, 87 and 3,5-dihydroxy-4-methoxybenzoic acid (88) have been isolated and identified using NMR spectroscopy. Phytochemical investigation of the bioactive butanol fraction of P. hydropiper using LC-DAD-ESIMS/MS led to the identification of nine compounds including rhamnazin-3-sulphate (91), quercetin-3-O-glucuronide (8), luteolin-3-O-glucoside (92), kaempferol-3-O-sulphate (93), quercetin-3-O-rhamnoside (7), apigenin-3-O-glucoside (89), quercitrin 7-sulphate (94), luteolin-7-sulphate (95) and isorhamnetin-3-sulphate (persicarin) (96). All compounds were relatively identified based on the MS/MS and UV data in comparison with those of the standard compounds. Antioxidants Persicaria Liquid chromatography 2011-06 Thesis http://psasir.upm.edu.my/id/eprint/27001/ http://psasir.upm.edu.my/id/eprint/27001/1/IB%202011%2018R.pdf application/pdf en public masters Universiti Putra Malaysia Antioxidants Persicaria Liquid chromatography Institute of Bioscience