Aldo Martelli

HPLC determination of caffeine and theophylline in Paullinia cupana Kunth (Guarana) and Cola spp. samples

ZusammenfassungDie Anwendung einer RP-HPLC für die mengenmässige Bestimmung von Coffein und Theophyllin in Guarana-Proben (Droge aus den Samen desPaullinia cupana-Baumgewächses Kunth, Familie:Sapindaceae) des Amazonas-Gebiet und in Cola spp.-Proben wird beschrieben. Die analytische Methode ist einfach, nicht zeitaufwendig und erfordert nur ein Mindestmaß an analytischen Operationen für die Proben-Zubereitung. Der Vergleich von 5 verschiedenen Extraktionstechniken für Coffein wird ebenfalls erörtert. Außerdem wurde der Gehalt von Coffein und Theophyllin in fünf verschiedenen brasilianischen Guarana-Proben, zwei Proben diätetischer Erzeugnisse mit Guarana-Gehalt, zwei Cola-Extraktund drei Cola-Samen-Proben bestimmt.SummaryA reversed-phase high-performance liquid-chromatographic method for the determination of caffeine and theophylline in commercial guarana samples (drug obtained from the seeds ofPaulinia cupana Kunth,Sapindaceae of the Amazon Region) and inCola spp. samples is described and discussed.The methodology developed is simple and rapid with a minimum of samples preparation required. A comparision of five different techniques for the extraction of caffeine and theophylline is discussed. Furthermore the quantitative determination of caffeine and theophylline in five samples of Brasilian guarana, in two samples of dietetic products containing guarana, in two samples ofCola extract and in three ofCola seed powder are reported.

Olive oil adulterated with hazelnut oils: simulation to identify possible risks to allergic consumers

According to European Union Regulation EC 1531/2001, olive oil labelled as “extra-virgin” should be cold-pressed and contain no refined oil or oil from other oleaginous seeds or nuts. Adulteration of extra virgin olive oil (EVOO) with hazelnut oil (HAO) is a serious concern both for oil suppliers and consumers. The high degree of similarity between the two fats complicates the detection of low percentages of HAO in EVOO. Many analytical approaches have been developed in recent years to trace HAO in EVOO, principally based on chromatographic analyses, differential scanning calorimetry or nuclear magnetic resonance. In addition adulteration of EVOO with HAO may introduce hazelnut-derived allergens. The aim of this work was to analyse the protein and allergen content of EVOO intentionally spiked with raw cold-pressed HAO or solvent-extracted HAO. SDS–PAGE analysis confirmed the presence of hazelnut proteins in solvent-extracted HAO with molecular masses ranging 10–60 kDa. In contrast, cold-pressed HAO showed no traces of protein. In spiked EVOO, solvent-extracted HAO was still detectable at a 1% contamination level. Several bands on SDS–PAGE migrated at apparent molecular masses coinciding with known allergens, such as Cor a 1 (∼17 kDa), Cor a 2 (∼14 kDa), Cor a 8 (∼12 kDa), oleosin (∼17 kDa) and Cor a 9 (∼60 kDa). MALDI–TOF MS analysis confirmed the presence of two oleosin isoforms and of Cor a 9. Immunoblotting demonstrated that an allergic patient with known reactivity to Cor a 1 and Cor a 2 recognized a 17-kDa band in solvent-extracted HAO. In conclusion, we have shown that adulteration of extra virgin olive oil with solvent-extracted hazelnut oil can be traced by simple SDS–PAGE analysis, and that adulteration introduces a potential risk for hazelnut allergic patients.

Protective activity of Theobroma cacao L. phenolic extract on AML12 and MLP29 liver cells by preventing apoptosis and inducing autophagy.

Theobroma cacao L. is known to have potential cardiovascular and cancer chemopreventive activities because of its high content of phenolic phytochemicals and their antioxidant capacities. In this work, we show for the first time that cocoa inhibits drug-triggered liver cytotoxicity by inducing autophagy. Phenolic-rich extracts of both unroasted and roasted cocoa prevented Celecoxib-induced cell viability inhibition in MLP29 liver cells because of the accumulation of G1 cells and cell death. Death prevented by cocoa had hallmarks of apoptosis such as the sub-G1 peak at flow cytometry and activation of Bax expression, together with down-regulation of Bcl-2, released cytochrome c in the cytosol with activation of Caspase 3, indicating that components of the apoptotic pathway such as Bax or upstream are major targets of cocoa phytochemicals. The protective effect of cocoa against liver cytotoxicity by Celecoxib was probably accounted for by inducing the autophagic process, as shown by enhanced Beclin 1 expression and accumulation of monodansylcadaverine in autolysosomes. This fact suggests that apoptosis was prevented by inducing autophagy. Finally, considering all these findings, we suggest that cocoa can be added to the list of natural chemopreventive agents whose potential in hepatopathy prevention and therapy should be evaluated.

Unusual essential oils with aromatic properties

ZusammenfassungEs wurde ätherisches Öl analysiert, das durch Wasserdampfdestillation von Chinarinde (Cinchona spp. derRubiaceae-Familie) gewonnen wurde. Diese Pflanze ist aufgrund ihrer pharmakologischen Eigenschaften bekannt und wird auch in der Lebensmittelindustrie verwendet, ganz besonders für die Likör- und Magenbitterherstellung. Die chemische Zusammensetzung dieses Öls war unbekannt. Nach Abtrennung auf einer Kieselgelsäule wurden im Öl mit GLC und GLC/MS etwa 100 Verbindungen identifiziert. Das Vorhandensein von 2-Hexyl-3-methyl-maleinsäureanhydrid ist ziemlich ungewöhnlich.SummaryThe essential oil obtained by steam distillation of cinchona bark (Cinchona spp.,Rubiaceae family) — a plant well known for its pharmacological properties, also employed in the food industry, especially in making liqueurs - was investigated. The oil, of completely unknown chemical composition, was analyzed by GLC and GLC/MS after separation on a silica gel column, and one hundred components were identified. The presence of 2-hexyl-3-methylmaleic anhydride is rather unusual.

Unusual essential oils with aromatic properties III. Volatile components of gentian roots

ZusammenfassungEs wurde etherisches Öl analysiert, das durch Wasserdampfdestillation aus Enzianwurzeln (Gentiana lutea L.,Gentianaceae) gewonnen wurde. Die Pflanze ist aufgrund ihrer pharmakologischen Eigenschaften bekannt und wird auch in der Lebensmittelindustrie verwendet, ganz besonders für die Likör-und Magenbitterherstellung. Das Öl wurde durch Säulenchromatographie an Kieselgel mittels GLC und GC/MS analysiert. Limonen (34,68%), Linalool, Carvacrol,cis-Linalyloxid und α-Terpineol bilden 50,04% des Öls. Darüber hinaus wurden weitere Bestandteile identifiziert.SummaryThe essential oil obtained by steam distillation of gentian roots (Gentiana lutea L.,Gentianaceae family) - a plant well know for its pharmacological properties and employed in the food industry, especially in making liqueurs - was investigated. The oil, after separation into fractions on a silica gel column, was investigated by GLC and GLC/MS. Limonene (34.68%), linalool, carvacrol, cis-linalyl oxide and aterpineol were the main components of the oil, totalling 50.04%. Several other minor components have been identified and are listed.