Development and validation of a UHPLC-ESI-MS/MS method for the quantification of artepillin C in Brazilian green propolis

Lorena Rodrigues Riani; LÍVIA MARA SILVA; OHANA OLIVEIRA ZUZA DA SILVA; LAURIENE RICARDO JUNQUEIRA; JORGE WILLIAN LEANDRO NASCIMENTO; ADEMAR ALVES DA SILVA FILHO

Lorena Rodrigues Riani Departamento de Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal de Juiz de Fora (UFJF), Juiz de Fora, MG, Brasil https://orcid.org/0000-0002-1847-2132
Lívia Mara Silva Departamento de Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal de Juiz de Fora (UFJF), Juiz de Fora, MG, Brasil
Ohana Oliveira Zuza da Silva Departamento de Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal de Juiz de Fora (UFJF), Juiz de Fora, MG, Brasil
Lauriene Ricardo Junqueira Departamento de Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal de Juiz de Fora (UFJF), Juiz de Fora, MG, Brasil
Jorge Willian Leandro Nascimento Departamento de Farmacologia, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora (UFJF), Juiz de Fora, MG, Brasil
Ademar Alves da Silva Filho Departamento de Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal de Juiz de Fora (UFJF), Juiz de Fora, MG, Brasil https://orcid.org/0000-0002-9096-1336

Keywords: Artepillin C, Brazilian Green Propolis, Liquid Chromatography, Mass Spectrometry., Quality Control

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.