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Title: HPLC-PDA isolation and LC-MS/MS detection of an acetylcholinesterase inhibitory flavonoid from Tephrosia purpurea (L.) Pers. in zebrafish brain
Authors: Arjun, Pitchai
Vincent, Samuel Gnana Prakash
Kannan, Rajaretinam Rajesh
Keywords: Acetylcholinesterase inhibition;Alzheimer’s disease;HPLC-PDA;HPLC purification;Wild indigo
Issue Date: Jun-2016
Publisher: NISCAIR-CSIR, India
Abstract: Separation of bioactive compounds or therapeutic small molecules from medicinal herbs is challenging due to the complexity of the phytochemicals. Tephrosia purpurea (L.) Pers. (Fam. Fabaceae) is rich in therapeutic compounds, used forisolation of an acetylcholinesterase inhibitor by HPLC coupled with Photo diode array (PDA) and mass spectrometric techniques. The separation was achieved through analytical HPLC by development of gradient mobile phase using methanol and acetonitrile along with 0.1 M ammonium acetate in Milli-Q water by two modes of mobile phase separations to yield the maximum purity, 99.13%. Based on the above-developed strategies, the preparative isolation of the acetylcholinesterase inhibitory flavonoid was purified at 269 nm with a retention time of 13. 9 min. The isolated compound from T. purpurea was confirmed as a flavonoid by phytochemical screening tests. The molecular mass was identified as 366.1467 Da by UPLC/Q-ToF-MS for the purified flavonoid.. Further, the molecular formula was found to be C22H22O5 by HR-MS/MS fragmentation pattern analysis through ChemSpider database search. The fragmentation pattern favoured the purified compound for similarity to Cyqualon. The isolated compound showed acetylcholinesterase (AchE) inhibition with the IC50 value of 54 µM in the zebrafish brain. However, the comparative study on the commercial cyqualon compound and the isolated compound showed different UV spectrum with the values at 269 nm and 257.5 nm, respectively. These findings concluded that the compound might be a novel flavonoid from Tephrosia purpurea which could be used as a therapeutic compound for neurodegenerative diseases after structural characterization.
Page(s): 104-111
ISSN: 0975-0959 (Online); 0301-1208 (Print)
Appears in Collections:IJBB Vol.53(3&4) [June-August 2016]

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