Abstract
The green alga Klebsormidium flaccidum var. zivo is a rich source of proteins, polyphenols, and bioactive small-molecule compounds. An approach involving chromatographic fractionation, anti-inflammatory activity testing, ultrahigh performance liquid chromatography-mass spectrometry profiling, chemometric analysis, and subsequent MS-oriented isolation was employed to rapidly identify its small-molecule anti-inflammatory compounds including hydroxylated fatty acids, chlorophyll-derived pheophorbides, carotenoids, and glycoglycerolipids. Pheophorbide a, which decreased intracellular nitric oxide production by inhibiting inducible nitric oxide synthase, was the most potent compound identified with an IC50 value of 0.24 µM in lipopolysaccharides-induced macrophages. It also inhibited nuclear factor kappaB activation with an IC50 value of 32.1 µM in phorbol 12-myristate 13-acetate-induced chondrocytes. Compared to conventional bioassay-guided fractionation, this approach is more efficient for rapid identification of multiple chemical classes of bioactive compounds from a complex natural product mixture.
Highlights
Classical approaches for identification of bioactive compounds from complex natural products involve time-consuming, multi-step bioassay-guided isolation procedures [1,2]
Using an approach involving chromatographic glycoglycerolipids, were identified to contribute to the anti-inflammatory activity of this alga. This fractionation, anti-inflammatory activity testing, ultrahigh performance liquid chromatography (UHPLC)-qMS profiling, chemometric analysis, and study highlights the utility of a chemometrics-assisted approach involving orthogonal partial least selective isolation and purification, four chemotypes of compounds including hydroxylated fatty acids, squares discriminant analysis (OPLS-DA) that can facilitate rapid identification of multiple bioactive chlorophyll-derived pheophorbides, carotenoids, and glycoglycerolipids, were identified to contribute compounds from a complex natural product mixture
Anti-inflammatory activity of extract determined in terms the decrease kappaB
Summary
Classical approaches for identification of bioactive compounds from complex natural products involve time-consuming, multi-step bioassay-guided isolation procedures [1,2]. KALGAETM ; and a 90-day dietary toxicity study of KALGAETM in CRL Sprague-Dawley CD IGS rats involving chromatographic fractionation, anti-inflammatory activity testing, UHPLC-qMS profiling, and a genotoxicity evaluation in Swiss albino mice did not show adverse effects, supporting its safe use chemometric analysis, and selective isolation and purification, four chemotypes of compounds as a potential food ingredient to improve human health [10]. Using an approach involving chromatographic glycoglycerolipids, were identified to contribute to the anti-inflammatory activity of this alga This fractionation, anti-inflammatory activity testing, UHPLC-qMS profiling, chemometric analysis, and study highlights the utility of a chemometrics-assisted approach involving orthogonal partial least selective isolation and purification, four chemotypes of compounds including hydroxylated fatty acids, squares discriminant analysis (OPLS-DA) that can facilitate rapid identification of multiple bioactive chlorophyll-derived pheophorbides, carotenoids, and glycoglycerolipids, were identified to contribute compounds from a complex natural product mixture. Results and Discussion rapid identification of multiple bioactive compounds from a complex natural product mixture
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