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

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, H NMR characteristic proton signals and polarity of compounds, eleven lipase inhibitors were tentatively assigned as triterpenoids: alisol B (m/z 495.50 [M + Na]), alisol B 23-acetate (m/z 537.58 [M + Na]), 11-deoxy-alisol B (m/z 479.50 [M + Na]), 11-deoxy-alisol B 23-acetate (m/z 521.50 [M + Na]), alisol A/epialisol A (m/z 513.50 [M + Na]), 16-oxo-11-deoxy-alisol A (m/z 511.50 [M + Na]), 16-oxo-alisol A (527.50 [M + Na] ), alisol C (m/z 509.58 [M + Na]), alisol C 23-acetate (m/z 551.50 [M + Na]), alisol M 23-acetate (m/z 567.50 [M + Na]), and alismanol Q/neoalisol (m/z 493.42 [M + Na]). 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.