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In this study, several spectroscopic and chromatographic techniques were employed to investigate the metabolic profile of different parts of Crocus sativus L. cultivated in Italy. The volatile profiling characterized by GC-MS for samples with the same genetic and geographical origin revealed some differences associated with different drying processes applied. LC-DAD/MS, NMR, FT-IR, and Raman were used to explore quality parameters of saffron , while other C. sativus flower parts (i.e. petals, leaves, and stamens) were also investigated. The metabolic profiling of saffron samples with different genetic origin but produced in the same region is also examined. C. sativus stamens, a possible bioadulterant, were studied to investigate potential marker compounds useful for their identification. All the results obtained highlight some aspects of genuine Italian saffron sold in Italy.

Saffron, the dried stigmas of the cultivated species Crocus sativus L., is the most valuable spice worldwide. The purpose of this short term scientific mission was to investigate the metabolic profile of different parts of Crocus sativus L. cultivated in Italy: in particular stigmas, stamens, tepals, and leaves were analysed. GC-MS analysis was used for the characterization of volatile compounds present in Italian saffron. The presence and the abundance of safranal and the HTTC, considered quality indices of saffron have been evaluated. The use of HPLC-DAD, FT-IR, and RAMAN explored: a) other quality parameters of saffron samples like crocins and picrocrocin; b) the metabolic profiling of saffron samples with different genetic origin but produced in the same region; c) other C. sativus tissues like: tepals for the presence of anthocyanins and flavonoids, leaves for the presence of phenolic compounds, and stamens, being a possible bioadulterant, were studied to look for potential marker compounds useful for their identification. All the results obtained highlighted some aspects of genuine Italian saffron of commercial grade (Zafferanami S.s.a.).

L'evento è nato nell'ambito del "Progetto Filagro: Strategie innovative e sostenibili per la filiera agroalimentare" finanziato nell'ambito dell'Accordo Quadro Regione Lombardia - Consiglio Nazionale delle Ricerche. FILAGRO intende dare risposte concrete alle esigenze emergenti nel settore dell'alimentazione e della produzione degli animali da latte. Lo studio sperimentale ha previsto che le diete delle capre fossero arricchite di semi di canapa e lino, con lo scopo di monitorare i cambiamenti nel contenuto di acidi grassi insaturi del latte, mediante analisi NMR, in quanto l'incremento nel contenuto di omega 3 e omega 6 migliora il valore nutraceutico del latte.

Flax (Linum usitatissimum L.) is a common fiber and oilseed crop considered as a valuable functional food since it constitutes one of the key sources of ?-3 fatty acids, phenylpropanoids (lignans) and mucilage [1]. During the last years there has been an increasing interest in dietary human consumption of flaxseeds in order to improve the nutritional and health status. In our laboratory we are interested in the study of phenylpropanoids metabolism. The main class of phenylpropanoids found in flax are lignans, diphenolic compounds displaying a broad range of biological activities such as antioxidant, cytotoxic, antifungal, antiviral, and phytoestrogenic [2; 3]. These phenolic compounds are of great interest for pharmaceutical, cosmetic and food industry. With the aim to identify and select the cultivar with the highest production of metabolites, the phenylpropanoid content of 7 flax cultivars (Linoal, Valoal, Natural, Festival, Merlin, Solal, Kaolin) and the expression level of key genes (PAL, Phenylalanine ammonia-lyase and 4CL, 4-Coumarate:CoA ligase) involved in their biosynthesis were compared. Moreover, the cultivar showing either the highest level of phenylpropanoids production and the highest level of expression of two key biosynthetic genes were chosen to start the production of flax cell cultures in order to improve the accumulation of bioactive compounds. In fact, it has been reported that plant cell cultures represent a promising system to optimize the production of valuable secondary metabolites [4]. Results. The 1H-NMR metabolite profiling of leaves (Fig.1) showed the presence of flavonoids, isoorientin, luteolin, apigenin and isovitexin, whereas seeds showed mainly lignans compounds. Real-Time PCR approach was used to evaluate the relative expression level of PAL and 4CL, representing two genes in the first part of phenylpropanoid biosynthetic pathway. Among the cultivars analysed, we found that Valoal and Solal are those with the highest expression level of PAL and 4CL both in leaves and in seeds (Fig.2). The determination of the specific activity of PAL and 4CL enzymes was in agreement with gene expression data, highlighting Valoal and Solal as the cultivars with the highest phenylpropanoids production. On the basis of these results Valoal was selected to start the production of in vitro cell cultures. To verify if in this system the ability to produce bioactive compounds was maintained we performed both biochemical and 1H-NMR analysis on the cultured cells. Preliminary analysis showed that flax cell cultures mainly produce compounds belonging to lignans, resembling the secondary metabolites identified in flax seeds. To increase the performance of flax cell cultures two strategies were used: a) the optimization of the medium composition and b) the use of some elicitors such as yeast extract, salicylic acid and methyl-jasmonate. Conclusion. The results obtained from flax leaves and seeds highlighted the presence of flavonoids and lignans with demonstrated antioxidant, antinflamatory, chemopreventive properties on human. The molecular analysis performed on the general phenylpropanoid pathway could help to better understand the biosynthesis of these compounds. Moreover, the preliminary results from the in vitro cultured cells suggest that this system is able to produce lignans as secondary metabolites resembling the flax seeds tissue. This finding is particularly encouraging since lignans have phytoestrogenic and cancer chemopreventive properties [5].