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      Identification and Quantification of Polyphenolic Secondary Metabolites in Stem Bark of Ficus religiosa (Moraceae) Using UPLC-HRMS and RP-HPLC-PDA

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          Abstract

          F. religiosa bark has been extensively used in traditional medicinal systems, such as Ayurveda, for its health benefits. The aim of this study was to investigate the secondary metabolites (phenolics and flavonoids) of the hydroalcoholic stem-bark extract from F. religiosa because this plant has been proven to have a beneficial effect on health disorders. Therefore, a pilot study was conducted for the identification and quantification of polyphenolic compounds in F. religiosa bark using sophisticated chromatographical techniques such as UPLC-HRMS and RP-HPLC-PDA. Additionally, total flavonoids, total phenolics and the scavenging profile of the bark were studied using a UV spectrophotometer. A total of 23 compounds identified with UPLC-HRMS were mainly phenolic acids, polyphenolics, and flavonoids (flavanols and proanthocyanidins). Among the identified compounds, gallic acid, catechin, epicatechin, epigallocatechin gallate, and ellagic acid were simultaneously quantified (0.031–0.380%) using RP-HPLC-PDA. Thereafter, the study complied by evaluating the total flavonoids (109.15 ± 1.2 mg RuE/g and 33.78 ± 0.86 mg CaE/g), total phenolics (4.81 ± 1.01 mg GaE/g), and scavenging profiles (IC50 13.75 ± 0.12 µg/mL) of the F. religiosa bark. This is the first report on the chemical profiling of F. religiosa bark, which is a necessary step to evaluate its nutraceutical properties, paving the way for possible food application.

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          Characterization of flavonoid subgroups and hydroxy substitution by HPLC-MS/MS.

          HPLC-DAD coupled with mass spectrometry in the positive ionization mode was applied to study the fragmentation of twelve selected flavonoids. Compounds belonging to all the major subgroups found in common plants, i.e. flavonols, flavones, dihydroflavonols, flavanones and flavanols were studied. Compound standards were injected into the spectrometer and produced characteristic mass spectra. The fragmentation of each compound was studied and it was shown that the dehydration and carbon monoxide losses from the [M+H]+ ion by the members of each subgroup produced specific fragments, thus allowing the characterization of the flavonoid subgroups. Moreover, fragments resulting from fission of the C-rings are specific of each subgroup and revealed the substitution pattern of A- and B-rings. In order to verify the identifying efficiency of the positive ionization mode through these characteristic fragmentations, the unknown flavonoids of an Origanum vulgare diethyl ether extract were separated with the HPLC system and the major peaks were successfully identified with the mass spectrometer.
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            Indigenous knowledge and uses of medicinal plants by local communities of the Kali Gandaki Watershed Area, Nepal.

            A field survey was conducted in the villages of Ramdi, Malunga, Balam, Beltari, Mirmi, Burgha and Ridi in the Kali Gandaki watershed, Nepal; 48 medicinal plants belonging to 31 families were reported, each with local names, traditional uses, methods of preparation and route of administration. Traditional medicine remains an integral part of the health system in these areas. Local people have remarkable knowledge of species identity and their uses as crude drugs.
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              Identification and characterization of chlorogenic acids, chlorogenic acid glycosides and flavonoids from Lonicera henryi L. (Caprifoliaceae) leaves by LC-MSn.

              The chlorogenic acids, chlorogenic acid glycosides and flavonoids of the leaves of Lonicera henryi L. (Caprifoliaceae) were investigated qualitatively by liquid chromatography tandem mass spectrometry. Thirty-one chlorogenic acids and their glycosides were detected and characterized to their regioisomeric level on the basis of their unique fragmentation pattern in the negative ion mode tandem MS spectra. All of them were extracted for the first time from this source and thirteen of them were not reported previously in nature. For the positive identification of chlorogenic acid glycosides by LC-MS(n), multiple reaction monitoring and targeted MS(n) experiments were performed. We have developed an LC-MS(n) method for the systematic identification of chlorogenic acid glycosides and were also able to discriminate between chlorogenic acids and their isobaric glycosides. It was also possible to discriminate between 5-O-(3'-O-caffeoyl glucosyl)quinic acid and 5-O-(4'-O-caffeoyl glucosyl)quinic acid by LC-MS(n). This method can be applied for the rapid and positive identification of chlorogenic acids and their glycosides in plant materials, food and beverages.
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                Author and article information

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                Journal
                SEPAF2
                Separations
                Separations
                2297-8739
                June 2023
                May 31 2023
                : 10
                : 6
                : 338
                Article
                10.3390/separations10060338
                fe4de898-c014-4e15-b1b9-49b2a71d24d4
                © 2023

                https://creativecommons.org/licenses/by/4.0/

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