Headspace with Gas Chromatography-Mass Spectrometry for the Use of Volatile Organic Compound Profile in Botanical Origin Authentication of Honey.

The botanical origin of honey determines its composition and hence properties and product quality. As a highly valued food product worldwide, assurance of the authenticity of honey is required to prevent potential fraud. In this work, the characterisation of Spanish honeys from 11 different botanical origins was carried out by headspace gas chromatography coupled with mass spectrometry (HS-GC-MS). A total of 27 volatile compounds were monitored, including aldehydes, alcohols, ketones, carboxylic acids, esters and monoterpenes. Samples were grouped into five categories of botanical origins: rosemary, orange blossom, albaida, thousand flower and "others" (the remaining origins studied, due to the limitation of samples available). Method validation was performed based on linearity and limits of detection and quantification, allowing the quantification of 21 compounds in the different honeys studied. Furthermore, an orthogonal partial least squares-discriminant analysis (OPLS-DA) chemometric model allowed the classification of honey into the five established categories, achieving a 100% and 91.67% classification and validation success rate, respectively. The application of the proposed methodology was tested by analysing 16 honey samples of unknown floral origin, classifying 4 as orange blossom, 4 as thousand flower and 8 as belonging to other botanical origins.

Hydrosols of orange blossom (Citrus aurantium), and rose flower (Rosa damascena and Rosa centifolia) support the growth of a heterogeneous spoilage microbiota.

Hydrosols are hydrodistillation products of aromatic plants. They contain less than 1g/L of dispersed essential oils giving organoleptic properties. Hydrosols are subjected to microbial proliferation. Reasons for spoilage have to be found in the nature of substrates supporting growth and of microbiological contaminants. The composition in essential oils and the microbiota of 22 hydrosol samples of Citrus aurantium L. ssp. amara L. (orange blossom), Rosa damascena Miller (rose D.), and Rosa centifolia L. (rose C.) flowers were analyzed to determine the factors responsible for decay. The median concentrations in essential oils were 677mg/L for orange blossom hydrosols, 205mg/L for rose D. hydrosols, and 116mg/L for rose C. hydrosols. The dry matter content of these hydrosols varied between 4.0mg/L and 702mg/L, and the carbohydrate content varied between 0.21mg/L and 0.38mg/L. These non-volatile compounds were likely carried over during distillation by a priming and foaming effect, and could be used as nutrients by microorganisms. A microbial proliferation at ambient temperature and also at 5°C has been observed in all studied hydrosols when stored in a non-sterile container. In contaminated hydrosols, maximal counts were about 7log10CFU/mL, while the French pharmacopeia recommends a maximal total bacterial count of 2log10CFU/mL. Neither yeast nor mold was detected. The isolated microbial population was composed of environmental Gram-negative bacteria, arranged in four major genera: Pseudomonas sp., Burkholderia cepacia complex, and presumably two new genera belonging to Acetobacteraceae and Rhodospirillaceae. Among those bacteria, Burkholderia vietnamiensis and Novosphingobium capsulatum were able to metabolize volatile compounds, such as geraniol to produce 6-methyl-5-hepten-2-one or geranic acid, or phenylethyl acetate to produce 2-phenylethanol. EO concentrations in hydrosols or cold storage are not sufficient to insure microbiological stability. Additional hurdles such as chemical preservatives or aseptic packaging will be necessary to insure microbial stability.

Development of new natural extracts.

For over the past 20 years, a remarkable development in the study and search of natural products has been observed. This is linked to a new market trend towards ecology and also due to new regulations. This could be a rupture, but also a real booster for creativity. Usually, in the flavor and fragrance field, creativity was boosted by the arrival of new synthetic molecules. Naturals remained the traditional, century-old products, protected by secrecy and specific know-how from each company. Regulatory restrictions or eco-friendly certification constraints like hexane-free processes triggered an important brainstorming in the industry. As a result, we developed new eco-friendly processes including supercritical CO2 extraction, allowing fresh plants to be used to obtain industrial flower extracts (Jasmine Grandiflorum, Jasmine Sambac, Orange blossom). These extracts are analyzed by GC, GC/MS, GCO, and HPTLC techniques. New or unusual raw materials can also be explored, but the resulting extracts have to be tested for safety reasons. Some examples are described.