Experimental Station for Essential Oils and Citrus Byproducts (SSEA) - Reggio Calabria (Italy)
dcautela@ssea.it
Laboratory Manager Fruit Juice and Vegetable Quality Control and Assurance Stazione Sperimentale per le Industrie delle Essenze e dei Derivati Dagli Agrumi -SSEA Italian Governmental Research Institute (Quality Assurance and Technological Innovation Research Area) (Reggio Calabria -Italy)
CURRENT RESEARCH ACTIVITIES AND INTERESTS: Nutritional quality of foods and bioavailability of food components. Detection of Persistent Organic Pollutants (PCBs, PCDDs/PCDFs) in food. New processes development for the phytochemicals recovery from fruit and vegetable byproducts. Food safety (heavy metals in food and food contact materials). Food chemistry (Antioxidants and phenolic compounds in vegetables).
Abstract: The occurrence of N-methylated tryptamine derivatives in bergamot plant (Citrus bergamia Risso et Poit) is reported for the first time. Interestingly, the most abundant of these substances is N,N,N-trimethyltryptamine, which has not been previously identified in any citrus plant. The N-methylated tryptamine derivatives were identified and quantitated in leaves, peel, juice, and seeds by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. N,N,N-Trimethyltryptamine was confirmed by MS(3) and comparison with the synthesized authentic standard. In addition, the study of the distribution of tryptophan, tryptamine, N-methyltryptamine, N,N-dimethyltryptamine, and N,N,N-trimethyltryptamine indicated that these compounds are differently expressed in the various tissues of the bergamot plant. Intriguingly, chemically synthesized N,N,N-trimethyltryptamine was reported to possess nicotine-like activity being a stimulant of parasympathetic ganglia by exerting its action on acetylcholine receptors. On this basis, the identification of N,N,N-trimethyltryptamine at a relatively high level in leaves suggests a possible role in a physiological mechanism of plant defense.
Abstract: The presence of pipecolic acid and pipecolic acid betaine, also known as homostachydrine, is herein reported for the first time in Citrus genus plants. Homostachydrine was found in fruits, seeds, and leaves of orange, lemon, and bergamot (Citrus bergamia Risso et Poit). As homostachydrine was not commercially available, as a comparative source, extracts of alfalfa leaves (Medicago sativa L.) were used, in which homostachydrine is present at high concentration. Then, the results where confirmed by comparison with an authentic standard synthesized and purified starting from pipecolic acid. The synthesized standard was characterized by a ESI-MS/MS study using a 3D ion-trap mass spectrometer. When subjected to MS/MS fragmentation in positive ion mode, homostachydrine, unlike its lower homologue proline betaine (also known as stachydrine), showed a pattern of numerous ionic fragments that allowed unambiguous identification of the compound. For the quantitation in the plant sources, high sensitivity and specificity were achieved by monitoring the transition (158â72), which is absent in the fragmentation patterns of other major osmolytes commonly used as markers for studies of abiotic stress. As for the metabolic origin of homostachydrine, the occurrence in citrus plants of pipecolic acid leads to the hypothesis that it could act as a homostachydrine precursor through direct methylation.
Abstract: he content of proline and various compounds deriving from its metabolism (4-hydroxy-L-proline, N-methyl-L-proline, N,N-dimethylproline, and 4-hydroxy-L-prolinebetaine) was determined in fruits and seeds of Bergamot (Citrus bergamia Risso et Poit), growing in the Calabria region (South Italy). A HPLC-ESI-tandem mass spectrometry method, which allowed rapid determination of L-proline, 4-hydroxy-L-proline, N-methyl-L-proline, N,N-dimethylproline, and 4-hydroxy-L-prolinebetaine in juice and extracts of bergamot fruit with minimum sample preparation and short analysis time (about 10 min), is presented. Proline and 4-hydroxy-L-proline levels in the samples were also determined by HPLC analysis with fluorescence detection and the results compared to those obtained with HPLC-ESI-tandem mass spectrometry. For the first time, the presence of N-methyl-L-proline and 4-hydroxy-L-prolinebetaine in the fruits of a plant of the Citrus genus is reported.
Abstract: Numerous compounds, many of them osmolytes, were quantified in natural juices and in frozen concentrate juices from fruits of plants of the Citrus genus. l-Proline, N-methyl-l-proline (hygric acid), N,N-dimethyl-l-proline (stachydrine), 4-hydroxy-l-prolinebetaine (betonicine), 4-hydroxy-l-proline, γ-aminobutyric acid (Gaba), 3-carboxypropyltrimethylammonium (GabaBet), N-methylnicotinic acid (trigonelline), and choline in the fruit juices of yellow orange, blood orange, lemon, mandarin, bitter orange ( Citrus aurantium ), chinotto ( Citrus myrtifolia ), and grapefruit were analyzed by sensitive HPLC-ESI-tandem mass spectrometry procedure. It was found that the most represented osmolytes in the juices, that is, l-proline, stachydrine, and betonicine, can be quantified with minimal sample preparation and short analysis time (about 1 min) also by flow injection analysis (FIA) ESI-MS/MS with the same results as obtained by HPLC ESI-MS/MS. In all of the juices, discrete amounts of choline and trigonelline were present. Conversely, GabaBet was always below detection limits. Notably, N-methyl-l-proline and 4-hydroxy-l-prolinebetaine, which were discovered for the first time in the juice of bergamot ( Citrus bergamia Risso et Poit), are also present in all of the citrus juices examined.
Abstract: The chemical composition of 30 samples of juices obtained from bergamot (Citrus bergamia Risso and Poit.) fruits is reported and compared to the genuineness parameters adopted by Association of the Industry of Juice and Nectars (AIJN) for lemon juice. It was found that the compositional differences between the two juices are distinguishable, although with difficulty. However, these differences are not strong enough to detect the fraudulent addition of bergamot juice to lemon juice. Instead, we found the high-performance liquid chromatography (HPLC) analysis of the flavanones naringin, neohesperidin, and neoeriocitrin, which are present in bergamot juice and practically absent in the lemon juice, is a convenient way to detect and quantify the fraudulent addition of bergamot juice. The method has been validated by calculating the detection and quantification limits according to Eurachem procedures. Employing neoeriocitrin (detection limit = 0.7 mg/L) and naringin (detection limit = 1 mg/L) as markers, it is possible to detect the addition of bergamot juice to lemon juice at the 1% level. When using neohesperidin as a marker (detection limit = 1 mg/L), the minimal percentage of detectable addition of bergamot juice was about 2%. Finally, it is reported that the pattern of flavonoid content of the bergamot juice is similar to those of chinotto (Citrus myrtifolia Raf) and bitter orange (Citrus aurantium L.) juices and that it is possible to distinguish the three kinds of juices by HPLC analysis.
Abstract: Investigation of PCDDs, PCDFs, PCBs, Al, As, Pb, Ba, Co, Cu, Cr, Fe, Mn, Cd, Ag, Sn, Zn, and Hg contents in 60 samples of cold-pressed essential oils produced in Calabria and Sicily in 2003-2005 was carried out. PCDDs, PCDFs, and PCBs were analyzed by HRGC-HRMS techniques using U.S. EPA 1613/94 and U.S. EPA 1668/A (1999) analytical methods. Mineral components were determined through GFAAS techniques; Hg content was determined by FI-M/H-AAS. The results of this study showed that essential oil contamination was due to a widespread pollution, typical background of rural areas, with relatively higher concentrations of PCDDs compared to PCDFs and little presence of PeCDF. Congeners OCDD, HpCDF, and OCDF were found at high concentrations. Regarding mineral components, mean values of Cr, Fe, and Ni were in agreement with data reported in the literature. Concentrations of As and Pb were below the maximum limits accepted by the current legislation. Finally, none of the samples analyzed were contaminated with Hg.
Abstract: A study on morphology, productive yield, main quality parameters and genetic variability of eight landraces of hot pepper (Capsicum annuum ssp.) from Southern Italy has been performed. Morphological characters of berries and productivity values were evaluated by agronomic analyses. Chemical and genetic investigations were performed by HPLC and random amplified polymorphic DNA (RAPD)-PCR, respectively. In particular, carotenoid and capsaicinoid (pungency) contents were considered as main quality parameters of hot pepper. For the eight selected samples, genetic similarity values were calculated from the generated RAPD fragments and a dendrogram of genetic similarity was constructed. All the eight landraces exhibited characteristic RAPD patterns that allowed their characterization. Agro-morphological and chemical determinations were found to be adequate for selection, but they resulted useful only for plants grown in the same environmental conditions. RAPD application may provide a more reliable way based on DNA identification. The results of our study led to the identification of three noteworthy populations, suitable for processing, which fitted into different clusters of the dendrogram.
Abstract: The mineral components distribution of cold pressed bergamot essential
oils was investigated by three different Atomic Spectrometry techniques.
Ag, Al, Ba, Be, Co, Cu, Cr, Fe, Mn, Ni, Tl, Sb, and Sn were quantified
by ICP-OES (Inductively Coupled Plasma Optical Emission
Spectrometry), while GFAA (Graphite Furnace Atomic Absorption) and
FI-M/H-AAS (Flow Injection Analysis-Mercury Hydride System) were
adopted for the determination of lead, cadmium and mercury.
Aluminium, tin and zinc were the elements with greatest concentration
present in the analyzed samples with an average content of 955, 582 and
418 ïg/Kg, while iron, barium and manganese assessed on average contents
of 269, 104 and 88 ïg/Kg.
Nickel and copper assessed on average contents of 17 and 29ïg/Kg,
while the average content of lead, manganese and zinc was 46, 88 e 418
ïg/Kg respectively
In all the analyzed samples the cobalt, beryllium and mercury concentrations
were result lower that limits of quantification, while the cadmium
was present to an average concentration of around 4 ïg/Kg.
Abstract: A procedure for the extraction and determination of pulegone enanthiomers in mint essential oils and mint products (syrups, dried leaves, toothpaste, lozenges, candy and chewing-gum) was developed. The compounds were recovered from the food matrices by employing a simultaneous distillation-extraction (SDE) technique with a Likens-Nickerson apparatus using dichloromethane as an extraction solvent. The analyses were performed by capillary gas chromatography mass spectrometry (GC/MS). Experiments on food products spiked at different pulegone concentrations showed recoveries ranging from 95 to 106%. The detection limit was about 5?mg?l(-1) for both pulegone enanthiomers and good linearity was found in the concentration range 0.5-25?mg?l(-1). In a number of repeated analyses, the pulegone peak height repeatability (RSD) was 0.2%. The pulegone enanthiomers were separated and quantified by enanthioselective multidimensional gas chromatography. The results of analyses conducted on essential mint oils and mint-flavoured food products are reported.
Abstract: The presence of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzo-p-furans (PCDFs) and polychlorinated biphenyls (PCBs) in 17 essential oil samples producted in Calabria and Sicily in 2003-2004 has been investigated. The analytical method used follows the European Directive 2002/69/EC guidelines. The extraction and clean up method resulted in an efficient separation of PCDD/Fs and PCBs from other interfering compounds. Analysis was carried out using an isotopic dilution method in conjunction with High Resolution Gas Chromatography-High Resolution Mass Spectrometry (HRGC-HRMS). Samples analysed as part of this survey show the presence of distinct PCDDs and PCDFs congeners like OCDD, HpCDF and OCDF in concentrations higher than the other congeners. The presence of non-2,3,7,8 chlorine substituted dioxins and furans congeners has been detected. High concentrations of PCBs congeners have also been found. As the essential oils industrial production processes don't involve any possible sources of contamination, the presence of PCDDs, PCDFs and PCBs found in the samples analysed might come from a diffuse environmental contamination.
Abstract: Pb, Ba, Co, Cu, Cr, Fe, Mn, Cd, Tl, Sb, Sn, Ag, Ni and Be in orange, lemon and mandarin "cold pressed" essential oils were analyzed by ICP-OES (Inductively Coupled Plasma Optical Emission Spectrometry) The best analytical conditions were assessed for each element and Detection Limits (DL), Limit of Quantitation (LOQ) and uncertainty were determined according to Eurachem guidelines. Tin was the element with greatest concentration present in the orange essential oils with an average content of 3,63 mg/Kg, followed by iron with an average content of around 1,43 mg/Kg and copper (1,37 mg/Kg). aluminium 0,82 mg/Kg and zinc with an average content of 0,82 mg/Kg. Regarding manganese and nickel content the average concentration values were respectively 0,31 and 0,29 mg/Kg. Barium and beryllium were not detected in the orange essential oils analyzed. In lemon essential oils, the most abundant element was the aluminium (average content 5,06 mg/Kg). Iron, tin, copper and zinc assessed with an average concentration of 2,33, 1,18, 1,04 and 0,66 mg/Kg, while manganese and nickel have shown an average concentrations around 0,19 mg/Kg. Barium and beryllium were not detected in the lemon essential oils analyzed Tin (3,49 mg/Kg) zinc (0.,14 mg/Kg) and nickel (0,18 mg/Kg) were results the most representative components in the mandarin essential oils. Lead, antimony and cadmium were generally not detected in the analyzed essential oils. Modest concentrations of barium and beryllium have been underlined in the analyzed mandarin essential oils. Cobalt and silver were respectively quantified only in one sample of orange essential oil and one sample of lemon essential oil (0,133 and 0,022 mg/Kg) and (0,108 and 0.036 mg/Kg), while were not detected in mandarin essential oil samples.
Abstract: A capillary zone electrophoresis (CZE) analitical method for the determination of ascorbic acid, potassium sorbate and sodium benzoate in fruit juices was validated by following EURACHEM guidelines and by comparing this method with International Federation of Fruit Juice Producers (IFFJP) ufficial methods. The method linear ranges were assessed in the ranges 5-500 mg/L for ascorbic acid and 5-200 mg/L for preservatives. The assessment of the method detection limits (MDL) and of the limits of quantitation (LOQ) were 0.1, 0.2, 0.6 mg/L and 0.5, 1, e 2 mg/L for potassium sorbate, ascorbic acid and sodium benzoate. The average spike recovery were 100â²5% for ascorbic acid and 97 â²4 % e 103â²5% for sodium benzoate and for potassium sorbate. A good correlation with ufficial methods has suggested that this method was suitable for the simultaneous determination of these three compound in frui and concentrated juices.
Abstract: The principal mineral components (Na, K, Ca, Mg) and trace elements (Al, As, Pb, Ba, Co, Cu, Cr, Fe, Mn, Cd, Se, Tl, Sb, Ag, Ni and Be) in Blood Orange and Chinotto juices were analyzed by ICP-OES (Inductively Coupled Plasma Optical Emission Spectrometry). The best analitical condiction were asseded for each element and Detection Limits (DL) were determined. The trace elements with greatest concentration present in chinotto juice were Iron, Boron and Zinc, with concentrations respectively of 1359, 1069 and 1424 μg/Kg. In the blood orange juice the concentrations of these trace elements were respectively 933, 2046 and 544 μg/Kg. Arsenic, Cadmium and Lead were not detected in the analyzed samples.
Abstract: The chemical composition of chinotto juice was analysed. Juice
yield, soluble solids, organic acids (citric acid, D-Isocitric acid
and Ascorbic acid), tritable acidity, sugars (glucose, fructose and
sucrose), metals content (Na, K, Ca, Mg), some trace elements,
free amino-acids and flavonoids (eriocitrin, neoeriocitrin, naringin
and hesperidin) were determined. Naringin is the main flavonoid
in the juice (average 800mg/L), followed by neohesperidin
(771 mg/L). Neoeriocitrin and eriocitrin content is respectively
of 467 and 33 mg/L
Abstract: A Glycosylated derivated of the neohesperidin dihydrochalcone, the 1-
(4- ((2- O- [6- deoxy- ï¡- L- mannopyranosyl]- 6- O- [ï¢- D- glucopyranosyl]-
ï¢- D- glucopyranosyl)oxy)- 2,6- dihydroxyphenyl)- 3- [3-
hydroxy- 4- methoxyphenyl]- 1- propanone, was prepared by precursory
isolations of flavonoid obtained from extraction of exhausted peels of the
workmanship of Citrus bergamia. From the neohesperidin contained in
the extract was prepared the 4'- metoxy- 3',5- di- O- acetil- flavanone- 7-
[2- O- (2,3,4- tri- O- acetyl- 6- deoxy- ï¡- L- mannopyranosyl)- 3,4- di-
O- acetyl- ï¢- D- glucopyranosyde], that was used as acceptor, with a reaction
of glycosylations that employed the 2,3,4,6- tetra- O- acetyl- ï¢- Dglucopyranosyl
bromide as donor. Glycosylations product, the 1- (4- ((2-
O- [6- deoxy- ï¡- L- mannopyranosyl]- 6- O- [ï¢- D- glucopyranosyl]-ï¢-
D- glucopyranosyl)oxy)- 2,6- dihydroxyphenyl)- 3- [3- hydroxy- 4-
methoxyphenyl]- 1- propanone, was purified and subsequently characterized
by thin layer chromatography (TLC), high pressure liquid chromatography
(HPLC), nuclear magnetic resonance (1H NMR) and elettrospray
mass spectrometry (ESI- MS). We showed that the reaction proceeds
in high yields. It can also be usefully employed for the preparation
of edulcorant substances, of discreet solubility in water and notable economic
interest, beginning from exhausted peels.
Abstract: The presence of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzo-p-furans (PCDFs) in buffalo milk and mozzarella cheese has been investigated. 64 cheese samples and 41 milk samples coming from Caserta region, south of Italy, have been analysed. The extraction and clean up method led an efficient separation of PCDD/F from other compounds that might interfere with the instrumental analysis. The isotopic dilution method has been used, and the instrumental analysis has been carried out by High Resolution Gas Chromatography-High Resolution Mass Spectrometry (HRGC-HRMS). Both these techniques applied make the analysis extremely selective and specific. The analytical results have been expressed as pg WHO-PCDD/F-TEQ/g fat, as requested by CE regulation CE N° 2375/2001, 29/1½001. Most of the samples analysed contain dioxins and furans congeners with higher molecular weighs but lower toxicity values. Therefore only five of the 41 milk samples and none of the 64 mozzarella cheese samples analysed show toxicity values higher than the law limits
