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Rapid identification of antioxidant compounds of Genista saharae Coss. & Dur. by combination of DPPH scavenging assay and HPTLC-MS.

ABSTRACT: Genista species are sources of antioxidant phenolic compounds such as O- and C-glycosyl?avonoids and isoflavonoids. A combination of a DPPH scavenging assay with HPTLC-MS, a fast and efficient method for identification of bioactive compounds, has been applied for evaluation of the radical scavenging activity of metabolites from Genista saharae Coss. & Dur. Different organs collected at various periods have been compared. Identification of antioxidant compounds was obtained by elution of the major DPPH-inhibition zones. The resulting HPTLC-MS analysis under moderately polar conditions, coupled to the DPPH results led to the putative identification of two antioxidant isoflavone aglycones: 3',4',5,7-tetrahydroxyisoflavone (1) and ficuisoflavone (3), whereas polar migration conditions led to the identification of the glycosides 5-methoxy-4',7-trihydroxy-8-glucopyranosylisoflavone (4) and 4',5-dihydroxy-7-methoxyisoflavone-4'-O-?-D-gluco-pyranoside (5). Evaluation of percentage of inhibition of DPPH radical by the purified isoflavone 4 from the root extract showed that it affords a moderate contribution to the total radical scavenging activity of the extract.

SUBMITTER: Meriane D

PROVIDER: S-EPMC6271480 | biostudies-literature | 2014 Apr

REPOSITORIES: biostudies-literature

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Rapid identification of antioxidant compounds of Genista saharae Coss. & Dur. by combination of DPPH scavenging assay and HPTLC-MS.

Meriane Djamila D Genta-Jouve Grégory G Kaabeche Mohamed M Michel Sylvie S Boutefnouchet Sabrina S

Molecules (Basel, Switzerland) 20140409 4

Genista species are sources of antioxidant phenolic compounds such as O- and C-glycosylﬂavonoids and isoflavonoids. A combination of a DPPH scavenging assay with HPTLC-MS, a fast and efficient method for identification of bioactive compounds, has been applied for evaluation of the radical scavenging activity of metabolites from Genista saharae Coss. & Dur. Different organs collected at various periods have been compared. Identification of antioxidant compounds was obtained by elution of the ...[more]

PMID: 24722590

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Project description:The objective of the present work was to develop a simple, specific, and fast high-performance thin-layer chromatographic (HPTLC) method to identify and quantify cordifolioside A, 20-β-hydroxyecdysone and columbin with HPTLC‒electrospray ionization‒tandem mass spectrometry (ESI‒MS/MS) for characterization in Tinospora cordifolia stem extracts. Chromatographic development was performed using a HPTLC aluminum plate, pre-coated with silica gel 60 F254 with hexane‒chloroform‒methanol‒formic acid as the mobile phase. Densitometric quantification for 20-β-hydroxyecdysone and cordifolioside A was performed at 254 nm and for columbin at 600 nm after derivatization with anisaldehyde‒sulfuric acid. The optimized mobile phase resulted in chromatographic separation of peaks for cordifolioside A, 20-β-hydroxyecdysone, and columbin at RF of 0.12, 0.47, and 0.86, respectively. The linear concentration range was found to be 750‒2250 ng/band for 20-β-hydroxyecdysone and cordifolioside A and 675‒1875 ng/band for columbin with (r2 > 0.99). The methodology showed good recoveries as 98.96‒101.43% for cordifolioside A, 98.15‒101.56% for 20-β-hydroxyecdysone, and 98.06‒98.80% for columbin. The limit of detection was found for columbin, 20-β-hydroxyecdysone, and cordifolioside A as 53.86 ng/band, 40.90 ng/band, and 107.05 ng/band, while the limit of quantification was found to be 163.21 ng/band, 123.94 ng/band, and 324.38 ng/band, respectively. The relative standard deviation for precision and robustness study for all the markers was found to be within 2%. Three markers were identified and confirmed in T. cordifolia stem extracts by ESI‒MS/MS. Compounds were assigned as norditerpene furan glycosides, ecdysteroids, and diterpenoid furanolactone: cordifolioside A (m/z = 527 [M + Na]+; UV λmax 221 nm), 20-β-hydroxyecdysone (m/z = 481.30 [M + H]+; UV λmax 247 nm), and columbin (m/z = 359 [M + H]+; UV λmax 210 nm). The optimized method was found accurate, reproducible, robust, and specific and can be applied for the quantification of cordifolioside A, 20-β-hydroxyecdysone, and columbin for quality control of extracts of T. cordifolia. Supplementary Information The online version contains supplementary material available at 10.1007/s00764-021-00115-7.

Dataset Information

Rapid identification of antioxidant compounds of Genista saharae Coss. & Dur. by combination of DPPH scavenging assay and HPTLC-MS.

Publications

Rapid identification of antioxidant compounds of Genista saharae Coss. & Dur. by combination of DPPH scavenging assay and HPTLC-MS.

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