Olfactory perception threshold assessment of volatile acidity in Madeira wines

Madeira wine is a fortified wine with impact in the Madeira Island’s economy. Similarly to other wines, its acidity should be well controlled in order to ensure Madeira wine quality, mostly the volatile acidity. Due to Madeira wine complex flavour, it is crucial to get a better knowledge about the volatile acidity impact in its features, namely determine the perception limit of acetic acid and ethyl acetate, as both are the main contributors for volatile acidity. Firstly, the olfactory perception threshold of volatile acidity was assessed by a trained and an untrained panel, using 5 and 10 years-old Sercial and Malvasia wines. Moreover, the current work also presents the evolution of organic acids, acetic acid and ethyl acetate during 540 days of ageing of Madeira wines (Malvasia, Bual, Verdelho and Sercial), comparing the same wines aged by both traditional ageing processes: canteiro and estufagem. Other wine samples, aged in wood casks (canteiro) for at least 5 years, were also evaluated. HS-SPME followed by GC-MS analysis was used to determine ethyl acetate concentration and IEC-HPLC-DAD was used for the organic acids determination, including acetic acid. The results indicated that acetic acid and ethyl acetate olfactory perception threshold depends essentially on wine’s age. Concerning acetic acid, the untrained panel was in average 5.45 g/L (5 years-old) and 6.22 g/L (10 years-old). Training the expert panel to recognize acetic acid odour, the values decreased for 1.44 g/L (5 years-old) and 1.87 g/L (10 years-old), but still remained higher than the established volatile acidity legal limits. Ethyl acetate threshold was similar for both panels (in average 327.97 mg/L). Both compounds tend to increase exponentially with age, being more evident in sweet wines. Organic acids in young Madeira wines depend mostly on the nature of grape varieties, but this difference is minimized with wine ageing.

Contribution to the chemoreception capacity of juvenile Loggerhead sea turtles (Caretta caretta, L.)

Loggerhead sea turtle juveniles (Caretta caretta), pelagic stage, are found in waters of Madeira archipelago. Pelagic turtles are in the main growth phase of their life cycle and consequently higher energy needs. However, knowledge about the ecology of pelagic loggerhead sea turtles is still quite rudimentary, mainly about the mechanisms that lead them to find food in the vast ocean. Studies with other pelagic species, such as procellariiform birds, revealed that the olfactory system play an important role for the detection of feeding areas, through the detection of concentration peaks of DMS (dimethylsulfide), a scent compound that naturally exists in the marine environment and it is related to areas of high productivity. Based on the assumption that loggerhead sea turtles use a similar mechanism, behavioural experiments were conducted in order to analyze the chemoreception capacity to DMS (airborne chemoreception - theoretically responsible for the long distance detection of areas with food patches; and aquatic chemoreception - theoretically responsible for the short distance detection of preys). The first step was to observe if pelagic loggerheads demonstrate sensitivity to DMS and the second was to verify if they really use the DMS, in natural conditions, as an airborne cue to find areas where food patches might be available. Four juveniles of loggerhead sea turtles were tested in captivity and three wild turtles in the open ocean. The results of airborne chemoreception experiments in captivity revealed that one turtle clearly demonstrated sensitivity to DMS and the sea experiments confirmed this result. However, the experiments were not conclusive on the question whether the pelagic turtles actually use the DMS as an airborne cue to detect long distance food patches. In aquatic chemoreception experiments was not observed sensitivity to DMS by the three sea turtles tested. In the classical conditioning experiment, where DMS and food were given nearly at the same time revealed that after a certain period of time, the sea turtle tested did not associated the DMS stimulus with a possible food reward. The main cause of mortality of loggerhead sea turtles in Madeira waters is due to the accidental capture (bycatch) by deep pelagic longlines fishery which the target species is the black-scabbard (Aphanopus carbo) fish. Chub mackerel (Scomber japonicus) is one of the baits used in this fishery. Aquatic chemoreception experiments were conducted in order to evaluate the attractiveness of the chub mackerel for sea turtles. For the three sea turtles tested, the results showed that in 90% of the cases the sea turtles were extremely attracted by the underwater smell of this fish.

Optimisation of stir bar sorptive extraction and liquid desorption combined with large volume injection-gas chromatography–quadrupole mass spectrometry for the determination of volatile compounds in wines

Stir bar sorptive extraction and liquid desorption followed by large volume injection coupled to gas chromatography–quadrupole mass spectrometry (SBSE–LD/LVI-GC–qMS) had been applied for the determination of volatiles in wines. The methodology was optimised in terms of extraction time and influence of ethanol in the matrix; LD conditions, and instrumental settings. The optimisation was carried out by using 10 standards representative of the main chemical families of wine, i.e. guaiazulene, E,E-farnesol, β-ionone, geranylacetone, ethyl decanoate, β-citronellol, 2-phenylethanol, linalool, hexyl acetate and hexanol. The methodology shows good linearity over the concentration range tested, with correlation coefficients higher than 0.9821, a good reproducibility was attained (8.9–17.8%), and low detection limits were achieved for nine volatile compounds (0.05–9.09 μg L−1), with the exception of 2-phenylethanol due to low recovery by SBSE. The analytical ability of the SBSE–LD/LVI-GC–qMS methodology was tested in real matrices, such as sparkling and table wines using analytical curves prepared by using the 10 standards where each one was applied to quantify the structurally related compounds. This methodology allowed, in a single run, the quantification of 67 wine volatiles at levels lower than their respective olfactory thresholds. The proposed methodology demonstrated to be easy to work-up, reliable, sensitive and with low sample requirement to monitor the volatile fraction of wine.