Characterization, quantitation, and natural variability of phenolics in Rhododendron arboreum leaves of western Himalayan origin using UPLC-PDA and UHPLC-ESI-Q/TOF-MS/MS

Rhododendron arboreum Sm. is very popular in the Indian Himalayan region due to its significant ethnomedicinal importance and potential commercial applications. Apart from the ornamental value derived from its vibrant red flowers, fresh flowers are used to prepare medicinal juices, and dried are utilized as spices. Despite being a rich source of phenolics, these lack identified marker chemicals and validated methods. The present work encompasses the chemical investigation of R. arboreum leaves using UHPLC-ESI-Q/TOF-MS/MS. The chemotypic variability was assessed through a reverse phase ultra-performance liquid chromatography method coupled with a photodiode array detector (RP-UPLC-PDA) chemical signatures of R. arboreum leaves from different altitudes combined with chemometric analysis. The chromatographic method was developed and validated for the quantitative analysis of three vital secondary metabolites, i.e., Isoquercetin or Quercetin-3-O-glucoside (Qg), Reynoutrin or Quercetin-3-O-xyloside (Qx), and Quercitrin or Quercetin-3-O-rhamnoside (Qr)The hyphenation of the method with ESI-Q/TOF-MS/MS has confirmed the distribution of 11 chemicals (nine known and two unknown) in R. arboreum leaves belonging to the phenolic and hydrolyzable tannins class. Samples from higher altitudes >3000 m (above mean sea level) were exceptionally rich in Quercitrin. The chemometric investigation, comprising hierarchical cluster analysis (HCA), principal component analysis (PCA), and partial least square method-discriminant analysis (PLS-DA), successfully categorized thirty-three samples from diverse geographic regions into two distinct categories. This current study provides a validated method for the chemotypic discrimination and quality assessment of the R. arboreum leaves from Uttarakhand, Western Himalayan region of India, and laid the foundation for the comprehensive utilization of these renewable and sustainable resources. [Display omitted] •Flavone glycosides (FOGs) and Quercitrin content of R. arboreum changed with elevations in Western Himalaya.•Isoquercitrin, Reynoutrin, and Quercitrin reported from Rhododendron arboreum leaves.•Validated UPLC-PDA provide rapid analysis of FOGs in R. arboreum leaves and its derived products.•Nine metabolites were putatively identified based on HRMS global fingerprinting.•Chemometric study demonstrate the similarities/dissimilarities among the population.