Metabolite Analysis of Toosendanin by an Ultra-High Performance Liquid Chromatography-Quadrupole-Time of Flight Mass Spectrometry Technique

Abstract

Toosendanin is the major bioactive component of Melia toosendan Sieb. et Zucc., which is traditionally used for treatment of abdominal pain and as an insecticide. Previous studies reported that toosendanin possesses hepatotoxicity, but the mechanism remains unknown. Its bioavailability in rats is low, which indicates the hepatotoxicity might be induced by its metabolites. In this connection, in the current study, we examined the metabolites obtained by incubating toosendanin with human live microsomes, and then six of these metabolites (M1-M6) were identified for the first time by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF/MS). Further analysis on the MS spectra showed M1, M2, and M3 are oxidative products and M6 is a dehydrogenation product, while M4 and M5 are oxidative and dehydrogenation products of toosendanin. Moreover, their possible structures were deduced from the MS/MS spectral features. Quantitative analysis demonstrated that M1-M5 levels rapidly increased and reached a plateau at 30 min, while M6 rapidly reached a maximal level at 20 min and then decreased slowly afterwards. These findings have provided valuable data not only for understanding the metabolic fate of toosendanin in liver microsomes, but also for elucidating the possible molecular mechanism of its hepatotoxicity.

Tribulus terrestris extracts alleviate muscle damage and promote anaerobic performance of trained male boxers and its mechanisms: Roles of androgen, IGF-1, and IGF binding protein-3

“…Q-TOF mass spectrometry can provide high resolution and accurate mass measurement of both the precursor and product ions, thus it has become more and more popular for identification of drug components and metabolites. 22 Twenty-two constituents were identified from the TT extracts, among which the most abundant constituents were TT saponins (25(R)-Spirostan-3,6,12-trione/25(R)-Spirostan-4-ene-3,12-dione and TT saponin A). The compositions and quantitative contents of TT extracts are not stable, depending on geographical region, climate 23 and part of herb, which may partly explain the divergent results of TT extracts from different studies.…”

The function of androgen/androgen receptor and insulin growth factor-1/insulin growth factor-1 receptor on the effects of Tribulus terrestris extracts in rats undergoing high intensity exercise

“…In contrast, TT saponins without any contamination do not lead to positive anti-doping tests (17). To exclude the possibility of AAS contamination, the chemical components of the TT extracts were analyzed by UHPLC-Q-TOF/MS, a popular method to identify medicine components and metabolites (35). Tigogenin and diosgenin, both saponins, were demonstrated account for ~71.35% of the total peak area, consistent with the promise of the manufacturers: >70% TT saponins.…”

“…The TT extract powder was dissolved in 70% ethanol and the supernatant was obtained by ultrasonic extraction and centrifugation at 20,217 x g for 10 min at 4˚C. The composition was determined using ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF/MS) revised according to the protocol of Wu et al (35) by Dr Xin Dong (College of Pharmacy, The Second Military Medical University, Shanghai, China).…”

Chemical Labeling Assisted Detection and Identification of Short Chain Fatty Acid Esters of Hydroxy Fatty Acid in Rat Colon and Cecum Contents

“…The loss of dimethylamino moiety is commonly observed for the DMED-labeled FAHFAs [ 16 ], and it was observed to be the most predominant ion for all the identified SFAHFAs. The product ion at m / z 406 was annotated as the loss of a neutral ketene or CH 2 CO ( m / z 42 Da) from the fragment ion m / z 448 [M+H–NH(CH 3 ) 2 ] + , which is normally observed for acetic acid derivatives [ 18 ]. Furthermore, the product ion at m / z 406 [M+H–NH(CH 3 ) 2 –CH 2 CO] + loses a neutral molecule of water ( m / z 18 Da) to afford a fragment ion at m / z 338 [M+H–NH(CH 3 ) 2 -CH 2 CO-H 2 O] + .…”