Optimization and validation of reverse phase HPLC method for qualitative and quantitative assessment of polyphenols in seaweed
Files in This Item:
|Manuscript_gaurav.doc||548.5 kB||Unknown||Download Request a copy|
|Title:||Optimization and validation of reverse phase HPLC method for qualitative and quantitative assessment of polyphenols in seaweed||Authors:||Rajauria, Gaurav||Permanent link:||http://hdl.handle.net/10197/9669||Date:||30-Jan-2018||Online since:||2019-03-25T12:25:31Z||Abstract:||A simple reverse phase-high performance liquid chromatography (RP–HPLC) coupled to a diode array detector (DAD) and negative ion electrospray mass spectrometer (ESI–MS) method was developed for simultaneous identification and quantification of phenolic antioxidants in seaweed. The proposed method was validated in terms of linearity, limits of detection (LOD), limits of quantification (LOQ), recovery and intermediate precision. The calibration curves were linear with correlation coefficient ranging from 0.9909 to 0.9997 while the values of LOD (0.26–0.82 mg/L), LOQ (0.77–2.50 mg/L), recovery (≥97.2%) and precision in terms of retention time (%RSD ≤2.27) and peak area (% RSD ≤5.11) were satisfactory. Brown seaweed Himanthalia elongata used in this study was extracted with 60% methanol and the crude extract was cleaned with SPE (Solid Phase Extraction) cartridge. HPLC-DAD-MS/MS analysis of the SPE fraction allowed the identification of 7 phenolic compounds comprising phlorotannins, hydroxybenzoic acid, hydroxycinnamic acid and flavonols subclasses of polyphenols. Quantitative analysis of these compounds revealed the presence of phloroglucinol (394.1 ± 4.33 μg/g), gallic acid (96.3 ± 3.12 μg/g), chlorogenic acid (38.8 ± 1.94 μg/g), caffeic acid (44.4 ± 2.72 μg/g), ferulic acid (17.6 ± 0.85 μg/g), myricetin (8.6 ± 0.85 μg/g) and quercetin (4.2 ± 0.15 μg/g), in the extract. The SPE fraction were tested for antioxidant capacity which were significantly (P < 0.05) higher (EC50; 14.5 ± 0.57 mg/g) than the ascorbic acid (EC50; 35.8 ± 0.59 mg/g) and the crude extract (EC50; 46.3 ± 0.48 mg/g). The occurrence of all these phenolic antioxidant compounds in H. elongata extract suggested that the developed method is sensitive enough and reproducible and could be used for qualitative and quantitative assessment of polyphenols in seaweed.||Type of material:||Journal Article||Publisher:||Elsevier||Journal:||Journal of Pharmaceutical and Biomedical Analysis||Volume:||148||Start page:||230||End page:||237||Copyright (published version):||2017 Elsevier||Keywords:||Phenolic antioxidant; Method optimization; Method validation; Seaweed; Solid phase extraction; LC-DAD-ESI–MS/MS; Electrospray mass spectrometer||DOI:||10.1016/j.jpba.2017.10.002||Language:||en||Status of Item:||Peer reviewed|
|Appears in Collections:||Agriculture and Food Science Research Collection|
Show full item record
This item is available under the Attribution-NonCommercial-NoDerivs 3.0 Ireland. No item may be reproduced for commercial purposes. For other possible restrictions on use please refer to the publisher's URL where this is made available, or to notes contained in the item itself. Other terms may apply.