Rapid screening for natural lipase inhibitors from Alisma orientale combining high-performance thin-layer chromatography-bioautography with mass spectrometry.

Abstract

Lipase inhibitors are an attractive class of hypolipidemic compounds, which inhibit the activity of human pancreatic lipase, thereby preventing the absorption of triglycerides in vivo. As a library of promising lead compounds for drug development, traditional Chinese medicine (TCM) has gained growing attention in quick discovery and identification of enzyme inhibitors of natural-origin. The purpose of this work was to discover unknown lipase inhibitors from Alisma orientale by the activity oriented analysis method thin-layer chromatography-bioautography, then use electrospray ionization mass spectrometry technology via the elution based TLC-MS interface to identify their structures. As a result, eleven natural lipase inhibitors from Alisma orientale extracts were identified based on molecular mass and fragment ions obtained by HPTLC-MS, and further confirmed by a series of complementary means including UV spectra, 1H NMR characteristic proton signals and polarity of compounds, eleven lipase inhibitors were tentatively assigned as triterpenoids: alisol B (m/z 495.50 [M\xa0+\xa0Na]+), alisol B 23-acetate (m/z 537.58 [M\xa0+\xa0Na]+), 11-deoxy-alisol B (m/z 479.50 [M\xa0+\xa0Na]+), 11-deoxy-alisol B 23-acetate (m/z 521.50 [M\xa0+\xa0Na]+), alisol A/epialisol A (m/z 513.50 [M\xa0+\xa0Na]+), 16-oxo-11-deoxy-alisol A (m/z 511.50 [M\xa0+\xa0Na]+), 16-oxo-alisol A (527.50 [M\xa0+\xa0Na] +), alisol C (m/z 509.58 [M\xa0+\xa0Na]+), alisol C 23-acetate (m/z 551.50 [M\xa0+\xa0Na]+), alisol M 23-acetate (m/z 567.50 [M\xa0+\xa0Na]+), and alismanol Q/neoalisol (m/z 493.42 [M\xa0+\xa0Na]+). The integrated approach is an efficient method for rapid screening lipase inhibitors from complex plant extracts and provides a reasonable and favorable basis for the identification and separation of other enzymatic system and other important compounds with therapeutic values.