*Article* **Evaluating the Presence and Contents of Phytochemicals in Honey Samples: Phenolic Compounds as Indicators to Identify Their Botanical Origin**

**Lua Vazquez 1, Daniel Armada 1, Maria Celeiro 1, Thierry Dagnac 2,\* and Maria Llompart 1,\***

1 CRETUS, Department of Analytical Chemistry, Nutrition and Food Science, Universidade de Santiago de Compostela, E-15782 Santiago de Compostela, Spain; lua.vazquez.ferreiro@usc.es(L.V.);daniel.armada.alvarez@usc.es(D.A.);

 maria.celeiro.montero@usc.es (M.C.) 2 Galician Agency for Food Quality-Agronomic and Agrarian Research Centre (AGACAL-CIAM), Unit of Food andFeedSafetyandOrganicContaminants,Apartado 10,E-15080ACoruña,Spain

**\*** Correspondence: thierry.dagnac@xunta.gal (T.D.); maria.llompart@usc.es (M.L.)

**Abstract:** Honey is a natural product well known for its beneficial properties. It contains phytochemicals, a wide class of nutraceuticals found in plants, including compounds with highly demonstrated antimicrobial and antioxidant capacities as phenolic compounds and flavonoids. The main goal of this work is the development of a miniaturized and environmentally friendly methodology to obtain the phenolic profile of Galician honeys (Northwest Spain) from different varieties such as honeydew, chestnut, eucalyptus, heather, blackberry and multi-floral. The total phenolic content (TPC) and antioxidant activity (AA) were also evaluated. As regards sample preparation, miniaturized vortex (VE) and ultrasound assisted extraction (UAE) employing aqueous-based solvents were performed. Individual quantification of 41 target phenolic compounds was carried out by liquid chromatographytandem mass spectrometry (LC-MS/MS). Results revealed the presence of 25 phenolic compounds in the 91 analyzed samples, reaching concentrations up to 252 μg g<sup>−</sup>1. Statistical tools such as analysis of variance (ANOVA) and principal component analysis (PCA) were employed to obtain models that allowed classifying the different honeys according to their botanical origin. Obtained results, based on TPC, AA and ∑phenolic compounds showed that significant differences appeared depending on the honey variety, being several of the identified phenol compounds being responsible of the main differentiation.

**Keywords:** honey; polyphenols; phenolic profile; total phenolic content; antioxidant activity; liquid chromatography; tandem mass spectrometry; principal components analysis
