Abstract
Objectives: The aim of this study was the development and validation of a fast method to quantify artepillin C in green propolis using ultra high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS). Methods: High purity (97.8%) artepillin C was isolated from green propolis using chromatography techniques. Quantification was performed using a C18(2.1 x 100 mm; 1.7 μm) column, gradient of water and methanol (with 0.01% formic acid) as mobile phase, at a flow rate of 0.4 mL/min and 45 ºC in temperature. A mass spectrometer operated in selected reaction monitoring mode to monitor the deprotonated molecular ion of artepillin C (m/z 299) > fragment ion (m/z 200.12). Several parameters such as specificity, linearity, limit of detection (LOD), limit of quantitation (LOQ), precision, accuracy, and robustness were determined. Results: The method was linear in the 50 – 400 μg/mL range (r2 = 0.9906), showing LOD = 10.79 μg/mL and LOQ = 32.70 μg/mL with satisfactory intra-day and inter-day precision with relative standard deviation (RSD %) of 1.9% and 3.4%, respectively. The accuracy showed recovery of 93-104%, the method was robust and artepillin C was quantified in green propolis at 6.51%. Conclusions: The proposed method showed advantages in comparison with other methods, such as short analysis time and high selectivity for artepillin C.
Highlights
Propolis is a natural resinous material collected by bees (Apis mellifera L.) from certain plants and used by them mainly to protect their hives[1]
The produced extract was concentrated under vacuum to furnish 30 g of a crude extract, which was chromatographed in a vacuum-liquid chromatography (VLC) system using silica gel (70-230 mesh) and chloroform/methanol mixtures as eluent, furnishing fifteen fractions (I-XV)
The chemical structure of the isolated compound was established as artepillin C by 1H- (500 MHz) and 13C- (125 MHz) Nuclear Magnetic Resonance (NMR; Bruker Avance III 500 MHz spectrometer) data analysis in comparison with literature data[4,25]
Summary
Propolis is a natural resinous material collected by bees (Apis mellifera L.) from certain plants and used by them mainly to protect their hives[1]. The southeastern Brazilian propolis, called as green propolis, has been used in Brazilian traditional medicine as an antimicrobial, anti-inflammatory, and to prevent diseases, such as gastrointestinal disorders[2]. Financial support: LRR and LMS – received scholarship from CAPES – Coordenação de Aperfeiçoamento de Nível Superior. Conflicts of interest: LRR, LMS, OOZS, LRJ, JWLN and AASF declare no conflict of interest. The study was carried out at Federal University of Juiz de Fora-UFJF, Juiz de Fora, Minas Gerais, Brazil Received on March 08, 2021.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have