Chemical profiling and simultaneous quantification of major bioactive constituents from Cocculus hirsutus by UPLC-QqQ-MS

Abstract

Cocculus hirsutus is a widely used herb in traditional systems of medicine for the treatment of various diseases. In the present study, five alkaloids (1–5), two flavonoids (6–7), one triterpenoid (8), and three steroids (9–10) were isolated from the roots of Cocculus hirsutus and further crude extract was analyzed by LC-Q-Tof-MS/MS in positive ionization mode leading to the identification of ten metabolites through comparison of exact molecular masses from their MS/MS spectra, mass fragmentation studies and with literature data. In addition, a method was developed and validated for the quantification of four bio-active compounds [Sinococuline (1), Magnoflorine (2), (E)-N-feruloyltyramine (3), and 20-Hydroxyecdysone (10)] using UPLC-QqQ-MS in multiple reaction monitoring (MRM) mode for the first time. The method has shown good linearity with correlation coefficients (r2) higher than 0.9916 for all four compounds. The intra- and inter-day precision were in the range of 0.3–6.1% and from 0.7% to 8.8%, respectively. The matrix effects of all the four analytes were found in the range of 94.7 ± 2.8–112.7 ± 3.7%. Overall, our study provides a reliable and rapid approach by hyphenated LC-MS/MS using high-resolution mass spectrometers for identification and quantification of bioactive constituents from the root extracts of Cocculus hirsutus.