Raquel Rodríguez-Solana

Use of hydrodistillation and headspace solid-phase microextraction to characterize the volatile composition of different hop cultivars.

BACKGROUND Hop cones, the immature inflorescences of the female plant of Humulus lupulus L., have been used for centuries to improve the flavor of beer and can be also used for a great variety of other products. Four samples of hop, belonging to three different cultivars (Nugget, Saaz and Perle), were studied in the present work. Headspace solid-phase microextraction and hydrodistillation techniques were used to obtain the volatile profiles of the samples. RESULTS Independent of the technique employed, over 40 volatile compounds were detected in the hop pellet samples (esters, monoterpenes, monoterpenoids, sesquiterpenes and sesquiterpenoids). Sesquiterpenes and sesquiterpenoids represented the majority of the total aromatic compounds. The main compounds for all cultivars were myrcene, β-caryophyllene and humulene, but the presence of high amounts of β-farnesene in Saaz cultivar was highlighted. CONCLUSION Both techniques were suitable for studying qualitatively the volatile composition of hop pellets, but some differences were shown when studying the proportion of the main constituents of the volatile profiles. Understanding these differences may help researchers design future studies to advise the industry how to exploit the potential of each hop cultivar.

Comparison of Soxhlet, accelerated solvent and supercritical fluid extraction techniques for volatile (GC-MS and GC/FID) and phenolic compounds (HPLC-ESI/MS/MS) from Lamiaceae species.

INTRODUCTION Plants from the Lamiaceae family have been known traditionally for their beneficial health-promoting properties, attributed to their anti-inflammatory, anaesthetic and anti-microbial effects. OBJECTIVE The purposes of this study was to characterise the essential oils from four Lamiaceae plants by applying different extraction techniques. METHODS Accelerated solvent (ASE), Soxhlet and supercritical fluid (SFE) extraction methods were compared for their efficiency in obtaining the essential oils from plants. The volatile compounds were identified by GC-MS and the main chemotype was quantified by GC with flame ionisation detection (FID). Phenolic compounds were identified and quantified by HPLC and electrospray ionisation (ESI) with MS/MS. RESULTS The essential oils Mentha piperita (ct. menthol/menthone), Rosmarinus officinalis L. (ct. eucalyptol/camphor) and Origanum vulgare (ct. carvacrol/thymol), whereas Thymus vulgaris L. was found to be a pure chemotype (ct. thymol). All three extracts also contained six phenolic compounds. The highest extraction yields were achieved by the Soxhlet and ASE techniques, with M. piperita and R. officinalis L. producing the highest concentrations of rosmarinic and carnosic acids. Finally, it was observed that M. piperita and O. vulgare produced the highest total phenolic content, whereas R. officinalis L. and T. vulgaris L. produced the highest anti-oxidant activity. CONCLUSION The ASE and Soxhlet extraction techniques presented the highest yields of volatile and phenolic compounds, showing their suitability to characterise the chemical profile of aromatic plants.

Production of vinyl derivatives from alkaline hydrolysates of corn cobs by recombinant Escherichia coli containing the phenolic acid decarboxylase from Lactobacillus plantarum CECT 748T

The enzyme PAD from Lactobacillus plantarum CECT 748T decarboxylates some cinnamic acids namely p-coumaric acid (p-CA), caffeic acid (CA), and ferulic acid (FA) into their corresponding 4-vinyl derivatives (4-VD): 4-vinyl phenol (4-VP), 4-vinyl catechol (4-VC), and 4-vinyl guaiacol (4-VG), respectively, which are valuable food additives mainly employed as flavouring agents. The gene encoding this enzyme was cloned and overexpressed in Escherichia coli. Recombinant E. coli cells overproducing L. plantarum PAD showed a preference to degrade mainly p-CA and CA. Sterilized liquors obtained after alkaline hydrolysis of corn cob or alkaline hydrolysis of the solid residue coming from acid hydrolysis of corn cob were employed as growth media in fermentations performed in shaker or bioreactor. The fermentative process allowed converting 2222.8 mg/L p-CA into 993.9 mg/L 4-VP. The process described here allowed the production with a high-yield of a valuable food additive from a by-product of the food industry.

Bioproduction of 4-vinylphenol from corn cob alkaline hydrolyzate in two-phase extractive fermentation using free or immobilized recombinant E. coli expressing pad gene

In situ extractive fermentation was used to produce 4-vinyl derivatives from hydroxycinnamic acids extracted from corn cobs by recombinant Escherichia coli cells expressing Lactobacillus plantarum phenolic acid descarboxylase (PAD) gene. This microorganism mainly produced 4-vinylphenol (4VP) from p-coumaric acid (p-CA). In the first study , we observed that the concentrations of 4VP are higher than 1g/L which had a negative impact on decarboxylation of p-CA to 4VP by recombinant E. coli cells. Because of this, and in order to improve the downstream process, a two-phase aqueous-organic solvent system was developed. The results of the extractive fermentation indicated that it was possible to use hydrolyzates as aqueous phase to bioproduce 4VP, and recover simultaneously the product in the organic phase containing hexane. The detoxification of pre-treated corn cob alkaline hydrolyzate improved 4VP production up to 1003.5mg/L after 24h fermentation (QP=41.813mg/Lh). Additionally, preliminary experiments using cells immobilized in calcium alginate showed to be a good system for the biotransform of p-CA to 4VP in extractive fermentation, although the process hindered partially the recovery of 4VP in the organic phase.

Optimisation of accelerated ageing of grape marc distillate on a micro‐scale process using a Box–Benhken design: influence of oak origin, fragment size and toast level on the composition of the final product

We are gratefulto the Spanish Ministry of Economy and Competitiveness for the financial support of this work (project CTQ2015-71436-C2-1-R), which has partial financial support from the FEDER funds of the European Union and to Laffort for providing the samples of oak fragments. Dr Jose Manuel Salgado was supported by the grant SFRH/BPD/84440/2012 from Fundacao para a Ciencia e Tecnologia–FCT, Portugal.

Optimization of the process of aromatic and medicinal plant maceration in grape marc distillates to obtain herbal liqueurs and spirits.

BACKGROUND Herbal liqueurs are alcoholic beverages produced by the maceration or distillation of aromatic and medicinal plants in alcohol, and are also highly valued for their medicinal properties. The process conditions, as well as the number and quantity of the plants employed, will have a great influence on the quality of the liqueur obtained. The aim of this research was to optimize these important variables. RESULTS A Box-Benhken experimental design was used to evaluate the independent variables: alcohol content, amount of plant and time during the experimental maceration of plants in grape marc distillate. Four plants were assessed, with the main compound of each plant representing the dependent variable evaluated with respect to following the evolution of the maceration process. Bisabolol oxide A in Matricaria recutita L., linalool in Coriander sativum L. and eucalyptol in Eucalyptus globulus Labill. were quantified using a gas chromatography-flame ionization detector. Glycyrrhizic acid in Glycyrrhiza glabra L was determined using a high-performance liquid chromatography-diode array detector. Other dependent variables were also evaluated: total phenolic content, color parameters and consumer preference (i.e. appearance). CONCLUSION The experimental designs allowed the selection of the optimal maceration conditions for each parameter, including the preference score of consumers: 70% (v/v) of ethanol, 40 g L-1 plant concentration and a maceration process of 3 weeks.