Quantitative and qualitative analysis of triterpenoid saponins in Achyranthes bidentata Blume via UHPLC-ELSD/UPLC-IM-Q-TOF-MS/MS analysis

[Display omitted] •A precise, accurate, sensitive and green UHPLC-ELSD method for the quantification of nine specialized metabolites.•Hydroxyecdysone (1) and momordin IIc (6) were found as the most abundant compounds in A. bidentata.•Principal component analysis revealed phytochemical distinction among A. bidentata seeds, roots and stem.•Thirty-four specialized metabolites annotated via UPLC-IM-Q-TOF-MS/MS based chemical profiling. Achyranthes bidentata Blume is a medicinal plant known for its anticoagulative, anti-osteoporosis, immunomodulatory, and neuroprotective properties. It has high trade value and is a repository of diverse triterpenoid saponins. However, it suffers from the issue of adulteration and substitution. There is a lack of analytical methods for the quality assessment, comparative quantification, and characterization of different morphological parts of A. bidentata. Hence, ultra-high performance liquid chromatography-evaporative light scattering detector (UHPLC-ELSD)-based analytical method was developed for the qualitative and quantitative estimation of nine compounds. The UHPLC-ELSD method exhibited excellent linearity (R2 ≥ 0.98) in the 15.62–1000 µg/mL range. Limit of detection and quantification were obtained within 3.90–15.62 µg/mL and 12.98–52.01 µg/mL range, respectively. The relative standard deviation (RSD) for inter-day and intra-day precision ranged from 2.1-5.6 % and 0.7–3.9 %, respectively. The recovery (%) ranged between 88.9–108.4 %. 20-Hydroxyecdysone (1) and momordin IIc (6) were the most abundant compounds and were quantified in the maximum amount in stem n-butanol fraction (52.70 mg/g) and seeds n-butanol fraction (145.26 mg/g), respectively. Ginsenoside Ro (4), achyranoside A (3), wogonin (8) and, emodin (9) were quantified as minor compounds in seeds and roots. Moreover, principal component analysis (PCA) analysis and chemical profiling performed using ultra-high-performance liquid chromatography-ion mobility-quadrupole time of flight tandem mass spectrometry (UPLC-IM-Q-TOF-MS/MS) technique led to the annotation of thirty-four compounds, mainly triterpenoid saponins. The green character of the developed analytical method was also assessed. The study marks the first attempt at the quantitative analysis of achyranoside A (2), achyranoside B (3), C-5 (5), momordin IIc 6, wogonin (8) and emodin (9) in the genus Achyranthes. The current study provides a comprehensive chemical profile and a robust UHPLC-ELSD-based quantifi