BRPI0802501A2 - process of identification, characterization and determination of the falsification of natural honeys from apis melifera and native bees - Google Patents

Abstract

IDENTIFICATION, CHARACTERIZATION AND DETERMINATION OF FALSIFICATION OF MEALS FROM APIS MELIFERA AND NATIVE BEES. The present invention describes a process for differentiating Apis melifera or native bee honeys, both of different blooms, and for determining their falsification by the addition of glucose syrup, caramel, invert sugar and vegetable oils by chemical markers. with matrix assisted laser desorption ionization and atmospheric pressure ionization techniques coupled with mass spectrometry.

Description

Report of the Invention Patent for "IDENTIFICATION PROCESS, CHARACTERIZATION AND DETERMINATION OF FAILURE OF NATIVE APIS MELIFERAE BEES"

FIELD OF INVENTION

The present technology brings a process for the differentiation and identification of honey bee or native Apis honey, regardless of the flora or region, quickly, simply and inexpensively.

It has application against tampering and / or counterfeiting demels in general.

BACKGROUND OF THE INVENTION

Honey is a sugary food produced by bees from flower nectar and saccharide exudates from the living parts of plants. Although it consists mainly of water and sugars, other substances present in small concentrations account for many of its characteristics such as its taste, aroma and antioxidant activity (Bastos et al., 2002; De Maria & Moreira, 2003; Gheldof et al. (2002; Gheldolf et al. 2003; Al-Mamary et al. 2002).

As a commodity, honey has limited availability and a relatively high price, encouraging its tampering. Generally this process is done by adding other carbohydrates, especially commercial sugars (disaccharides) such as glucose or glucose syrup, melates or inverted desaccharose solution and even oils. vegetables.

Most of Brazil's honey production comes from A. meliferous bees brought from the European continent around 1839 and a small part is produced by native bees, mainly jatai bees (Tetragonisca angustuld). The honeys produced by A. melifer and T. angustula have characteristics different from each other and in terms of their vegetal origin, that is, deforestation, citrus, eucalyptus and others.

The characterization of honeys is based on microscopic pollen analysis and the physicochemical characteristics: shrinkage index, humidity, density, dynamic viscosity, pH, specific rotation, electrical conductivity, ashes, sugars, deproline content and phenolic compounds.

SUMMARY OF THE INVENTION

The process described here aims to differentiate the types of honeys through the chemical markers found in different relationships, both in the authentic honeys of A. honeyifer and native bees, both of different flowering, adulterated honeys quantons, without the need for microscopic pollen analysis. physicochemical characteristics using ionization techniques: matrix assisted laser desorption (MALDI), electrospray desorption (DESI), electrospray desorption (DESSI), real-time direct analysis (DART), electrospray (ESI), chemical ionization atmospheric pressure (APCI) and / or atmospheric pressure photoionization (APPI) coupled with mass spectrometry.

The advantages of using the methodologies developed in the present invention over the techniques usually employed are: in their simplicity, due to the minimum sample size; fast because it has an efficient data acquisition system; and sensitivity, characteristic of the mass spectrometry technique. Thus, there is great potential for future application on a commercial scale.

DETAILED DESCRIPTION OF THE INVENTION

For a better understanding of the present method, the differentiation of honey bee and native Apis bees by diagnostic ions which are objects of this patent consist of the following steps:

a) mixing and homogenizing honey and matrix (2,5-dihydroxybenzoic acid, dithranol, 3-hydroxy-2-pyridinecarboxylic acid, α-cyano-4-hydroxycinnamic acid, retinoic acid, cyclodextrin, or dithiothreitol) with mortar and pistil , analysis of the chemical constituents present in these mixtures using tinted laser desorption ionization mass spectrometry (MALDI-MS).

b) in situ analysis of the chemical constituents present by using electrospray desorption ionization mass spectrometry (DESI-MS), electrospray desorption mass spectrometry (DESSI-MS) and / or direct real time analysis (DART-MS).

c) dilution of samples in 1: 1 methanol / water or 1: 1 acetonitrile / water solution with 0.1% formic acid for the positive, or 0.1% ammonium hydroxide for the negative mode for analysis of the constituents of the using electrospray ionization mass spectrometry (ESI-MS), atmospheric pressure chemical ionization (APCI-MS) and / or chloroform / methanol 1: 1 with 0.1% formic acid for the modopositive, or 0.1% ammonium hydroxide for the negative mode for atmospheric pressure photoionization ionization coupled to mass spectrometry (APPI-MS) and / or electrosonicspray desorption (DESSI-MS).

With the proposed process, an experimental procedure was performed for the differentiation of honey from native A. meliferae bees of different flowers, in a commercial laboratory scale, of the different types of cultures, which is described in Example I. Example I:

A mixture of dihydroxybenzoic acid and honey matrix in the ratio of 5: 1 was homogenized in a pistil mortar, then in a ball mill and then applied to a previously cleaned and dried sample plate with the aid of a spatula. The chemical profile of the samples was analyzed, mainly observing the diagnostic ions of the different demel types by matrix assisted desorption ionization mass spectrometry (MALDI-MS). The main characteristics of the different types of honeys are related to sugars, triglycerides, tocopherols and minor polar secondary metabolites, where through direct analysis by matrix-assisted laser desorption ionization (MALDI), or by chemometric analysis, it was possible to differentiate. quickly and safely different types of honeys and possible tampering.

Claims (6)

1. Process for the identification, characterization and determination of the falsification of natural honeys from apis melifera and native bees, CHARACTERIZED by the analysis through chemical markers with ionization techniques. 2. Process for identifying, characterizing and determining the falsification of natural honeys according to claim 1, characterized by the fact that an ionization technique is by matrix assisted laser desorption. 3. Process for identifying, characterizing and determining the falsification of natural honeys according to claim 1, characterized by the fact that an ionization technique is by ionization at atmospheric pressure. 4. Process for the identification, characterization and determination of the falsification of natural honeys according to claims 2 and 3, characterized by the fact that ionization techniques are coupled to mass spectrometry. 5. Process for the identification, characterization and determination of falsification of natural honeys according to claim 2 or 3, characterized by the fact that matrix assisted laser desorption ionization (MALDI), electrospray desorption (DESI), deselection by electronicsrosonicspray apply. (DESSI), direct real time analysis (DART), electrospray (ESI), atmospheric pressure chemical ionization (APCI) and / or atmospheric pressure photoionization (APPI). 6. Process for the identification, characterization and determination of falsification of natural honeys according to claim 5, characterized in that the matrices used for the MALDI technique are 2,5-dihydroxybenzoic acid, dithranol, 3-hydroxy-2-pyridinecarboxylic acid. α-cyano-4-hydroxycinnamic acid, retinoic acid, cyclodextrin, or dithiothreitol.