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

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 C 18 (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 (r 2 = 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.

Comparison of the Biological Potential and Chemical Composition of Brazilian and Mexican Propolis

“…Many of these chemical compounds stand out for their biological activity, for example, phenolic acids, phenolic acid esters, flavonoids, terpenoids, luteolin, galantine, trans ferulic acid, caffeic acid, chrysin, pinobanksin 5-methyl ether, quercetin, apigenin, kaempferol, naringenin, rutin, catechin, p-coumaric, pinobanksin, pinocembrin, and pinobanksin 3-acetate, and many others [21,[25][26][27][28][29][30][31][32]. On the other hand, some compounds that are considered propolis-type markers, such as in propolis from Brazil, are artepillin C (3,5-diphenyl-4-hydroxycinnamic acid) for green propolis [44][45][46][47] and formononetin (3-hydroxy-8-9-dimethoxypterocarpan) for red propolis [32,[48][49][50]. These chemical markers described for each propolis are phytologically associated with the main botanical source.…”