Francesca Giuffrida

Identification of the botanical origin of pine nuts found in food products by gas-liquid chromatography analysis of fatty acid profile.

Pine nuts are traditionally used in various part of the world for the preparation of desserts or sauces or in salads. Local production is not sufficient to cope with the high demand of pine nuts around the world, and countries such as China or Pakistan are exporting much of their production to Western countries. Almost all the nuts that are traditionally consumed belong to the Pinus genus, but over the past years, the number of consumer complaints following consumption of commercial pine nuts increased. Some consumers experienced taste disturbance lasting for up to two weeks after consumption. Food safety agencies raised some concerns regarding pine nuts imported from Asia and their association with taste disturbance. However, even though a formal association has not been found to date, the Pinus genus comprises species that are not classified as edible and could be eventually used to adulterate edible species. Pinus spp. seed lipids are known to contain very specific polyunsaturated fatty acids know as Delta5-olefinic acids. Seed fatty acid profile of conifers had been used in the past as a taxonomic marker, and in the present study to identify the botanical origin of pine nut in nine commercial products. Fast gas-liquid chromatography (GLC) was used to resolve the complete fatty acid profile of Pinus spp. samples in less than 5 min. A diagnostic index based on the relative levels of the main fatty acids including distinctive Delta5-olefinic acids was used to identify botanical origins. Results revealed the occurrence of the following Pinus spp. in commercial products: P. pinea, P. koraiensis, P. gerardiana, P. armandii and P. massoniana. The later two species, known as Chinese white pine and Chinese red pine, are only cultivated in China and are not listed as common source of edible pine nuts by the Food and Agriculture Organization (FAO). The present study shows that the botanical origin of pine nuts can be identified in products based on the fatty acid profile.

Dynamics of human milk nutrient composition of women from singapore with a special focus on lipids

A recent report suggested that human milk (HM) composition not only changes with lactation stages but also vary according to gender of the offspring. In spite of available literature, the dynamic changes of HM composition still remain to be completely explored and characterized. Progress in analytical technologies together with quantitative sampling of HM allows for a better quantification of HM nutrients and thereby providing a deeper understanding of the dynamics of HM secretion.

Antioxidant Activity of Oregano, Parsley, and Olive Mill Wastewaters in Bulk Oils and Oil-in-Water Emulsions Enriched in Fish Oil

The antioxidant activity of oregano, parsley, olive mill wastewaters (OMWW), Trolox, and ethylenediaminetetraacetic acid (EDTA) was evaluated in bulk oils and oil-in-water (o/w) emulsions enriched with 5% tuna oil by monitoring the formation of hydroperoxides, hexanal, and t-t-2,4-heptadienal in samples stored at 37 degrees C for 14 days. In bulk oil, the order of antioxidant activity was, in decreasing order (p < 0.05), OMWW > oregano > parsley > EDTA > Trolox. The antioxidant activity in o/w emulsion followed the same order except that EDTA was as efficient an antioxidant as OMWW. In addition, the total phenolic content, the radical scavenging properties, the reducing capacity, and the iron chelating activity of OMWW, parsley, and oregano extracts were determined by the Folin-Ciocalteau, oxygen radical absorbance capacity, ferric reducing antioxidant power, and iron(II) chelating activity assays, respectively. The antioxidant activity of OMWW, parsley, and oregano in food systems was related to their total phenolic content and radical scavenging capacity but not to their ability to chelate iron in vitro. OMWW was identified as a promising source of antioxidants to retard lipid oxidation in fish oil-enriched food products.

High-throughput methods to assess lipophilic and hydrophilic antioxidant capacity of food extracts in vitro.

Assays comprising three probes for different mechanisms of antioxidant activity in food products have been modified to allow better comparison of the contributions of the different mechanisms to antioxidant capacity (AOC). Incorporation of a common format for oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), and iron(II) chelating activity (ICA) assays using 96-well microplates provides a comprehensive and high-throughput assessment of the antioxidant capacity of food extracts. The methods have been optimized for aqueous extracts and validated in terms of limit of quantification (LoQ), linearity, and precision (repeatability and intermediate reproducibility). In addition, FRAP and ORAC assays have been validated to assess AOC for lipophilic extracts. The relative standard deviation of repeatability of the methods ranges from 1.2 to 6.9%, which is generally considered to be acceptable for analytical measurement of AOC by in vitro methods. Radical scavenging capacity, reducing capacity, and iron chelating properties of olive mill wastewaters (OMWW), oregano, and parsley were assessed using the validated methods. OMWW showed the highest radical scavenging and reducing capacities, determined by ORAC and FRAP assays, respectively, followed by oregano and parsley. The ability to chelate Fe (2+) was, in decreasing order of activity ( p > 0.05) parsley congruent with oregano > OMWW. Total phenol content, determined by the Folin-Ciocalteu method, correlated to the radical scavenging and reducing capacities of the samples but not to their chelating properties. Results showed that the optimized high-throughput methods provided a comprehensive and precise determination of the AOC of lipophilic and hydrophilic food extracts in vitro.

Dose-response plasma appearance of coffee chlorogenic and phenolic acids in adults.

SCOPE Coffee contains phenolic compounds, mainly chlorogenic acids (CGAs). Even though coffee intake has been associated with some health benefits in epidemiological studies, the bioavailability of coffee phenolics is not fully understood. OBJECTIVE AND STUDY DESIGN We performed a dose-response study measuring plasma bioavailability of phenolics after drinking three increasing, but still nutritionally relevant doses of instant pure soluble coffee. The study design was a one treatment (coffee) three-dose randomized cross-over design, with a washout period of 2 wks between visits. RESULTS CGAs, phenolic acids, and late-appearing metabolites all increased with increasing ingested dose. Hence, the sum of area under the curve was significantly higher for the medium to low dose, and high to medium dose, by 2.23- and 2.38-fold, respectively. CGAs were not well absorbed in their intact form, regardless of the dose. CGA and phenolic acids appeared rapidly in plasma, indicating an early absorption in the gastrointestinal tract. Late-appearing metabolites were the most abundant, regardless of the dose. CONCLUSION This study confirmed previous findings about coffee bioavailability but also showed that coffee phenolics appear in a positive dose-response manner in plasma when drank at nutritionally relevant doses.

UPLC-MS/MS quantification of total hesperetin and hesperetin enantiomers in biological matrices.

Hesperidin (hesperetin-7-O-rutinoside), a flavonoid affecting vascular function, is abundant in citrus fruits and derived products such as juices. After oral administration, hesperidin is hydrolyzed by the colonic microbiota producing hesperetin-7-O-glucoside, the glucoside group is further cleaved and the resulting hesperetin is absorbed and metabolized. Flavanones have a chiral carbon generating (R)- and (S)-enantiomers, with potentially different biological activities. A rapid UPLC-MS/MS method for the analysis of (R)- and (S)-hesperetin enantiomers in human plasma and urine was developed and validated. Biological matrices were incubated with β-glucuronidase/sulfatase, and hesperetin was isolated by solid-phase extraction using 96-well plate mixed-mode cartridges having reversed-phase and anion-exchange functionalities. Racemic hesperetin was analyzed with a UPLC HSS T3 reversed phase column and hesperetin enantiomers with a HPLC Chiralpak IA-3 column using H(2)O with 0.1% CHOOH as solvent A and acetonitrile with 0.1% CHOOH as solvent B. The method was linear between 50 and 5000nM for racemic hesperetin in plasma and between 25 and 2500nM for (S)- and (R)-hesperetin in plasma. Linearity was achieved between 100 and 10,000nM for racemic hesperetin in urine and between 50 and 5000nM for (S)- and (R)-hesperetin in urine. Values of repeatability and intermediate reproducibility for racemic hesperetin and enantiomers in plasma and urine were below 15% of deviation in general, and maximum 20% for the lowest concentrations. In addition, the method was applied for the quantification of total hesperetin and of hesperetin enantiomers in human plasma and urine samples, obtained after oral ingestion of purified hesperetin-7-O-glucoside. In conclusion, the developed and validated method was sensitive, accurate and precise for the quantification of enantiomers of hesperetin in biological fluids.

Quantification of phenolic acids and their methylates, glucuronides, sulfates and lactones metabolites in human plasma by LC–MS/MS after oral ingestion of soluble coffee

Chlorogenic acids and derivatives like phenolic acids are potentially bioactive phenolics, which are commonly found in many foods. Once absorbed, chlorogenic and phenolic acids are highly metabolized by the intestine and the liver, producing glucuronidated and/or sulphated compounds. These metabolites were analyzed in human plasma using a validated liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method. After protein precipitation, phenolic acids and their metabolites were extracted by using ethanol and chromatographic separation was achieved by reversed-phase using an Acquity UPLC BEH C18 column combined with a gradient elution system using 1% acetic acid aqueous solution and 1% acetic acid with 100% acetonitrile. The method was able to quantify 56 different compounds including 24 phenolic acids, 4 lactones, 15 sulfates and 13 glucuronides metabolites between 5 and 1000nM in plasma for most of them, except for m-dihydrocoumaric acid, 5-ferulloylquinic-glucuronide, 4-methoxycinnamic acid, 3-phenylpropionic acid, 3-(4-methoxyphenyl)propionic acid (25 to 1000nM) and p-dihydrocoumaric acid (50-1000nM). Values of repeatability and intermediate reproducibility were below 15% of deviation in general, and maximum 20% for the lowest concentrations. The validated method was successfully applied to quantify phenolic acids and their metabolites in plasma obtained after oral ingestion of soluble coffee. In conclusion, the developed and validated method is proved to be very sensitive, accurate and precise for the quantification of these possible dietary phenols.

Oxidative Stability at High Temperatures of Oleyol and Linoleoyl Residues in the Forms of Phosphatidylcholines and Triacylglycerols

An investigation was carried out into the stability of fatty acyl groups to heat-induced oxidative changes as affected by their chemical environment. The behavior of oleic and linoleic acyl groups when esterified in triacylglycerols (TAGs) and phosphatidylcholines (PCs) was evaluated. The monitoring of the oxidative degradation using liquid chromatography-mass spectrometry (LC-MS) showed that fatty acyl groups are less likely to be oxidized when in the form of PCs than when in the form of TAGs. In addition, oxidation products from PCs were more stable than those from TAGs. This finding strengthens the idea that the choline group in PCs increases the stability of fatty acyl groups to oxidation in comparison to TAGs.

In vitro digestion of citric acid esters of mono- and diglycerides (CITREM) and CITREM-containing infant formula/emulsions

CITREM is an emulsifier used in the food industry and contains citric acid esters of mono- and diglycerides (GCFE). It is generally recognized as safe but no publication on its digestibility under gastrointestinal conditions and impact on fat digestion was available. It was shown here that fatty acids are released from CITREM by gastric lipase, pancreatic lipase, pancreatic-lipase-related protein 2 and carboxyl ester hydrolase. A two-step in vitro digestion model mimicking lipolysis in the stomach and upper small intestine of term and preterm infants was then used to evaluate the digestibility of CITREM alone, CITREM-containing infant formula and fat emulsions, and isolated GCFE fractions. Overall, it was shown that fat digestion is not significantly changed by the presence of CITREM, and fatty acids contained in CITREM compounds are released to a large extent by lipases. Nevertheless, undigestible water-soluble compounds containing glycerol and citric acid units were identified, indicating that the ester bond between citric acid and glycerol is not fully hydrolyzed throughout the proposed digestion.

Identification of the botanical origin of commercial pine nuts responsible for dysgeusia by gas-liquid chromatography analysis of Fatty Acid profile.

Over the last 10 years, complaints were increasingly reported from consumers that experienced dysgeusia following the consumption of pine nuts. In the present study, pine nuts samples (N = 16) from consumers that reported dysgeusia have been analyzed to identify the botanical origin of critical pine nuts samples. The fatty acid composition of the samples was performed, and diagnostic index values were used to identify the botanical origin of the samples. Pinus armandii nuts were identified in all the samples pure or in mixture with P. koraiensis nuts. P. armandii is not reported as edible pine nuts by the Food and Agriculture Organization (FAO). This study confirmed that consumption of P. armandii nuts may lead to dysgeusia. Based on the present study and previous work, we advise import companies to trade pine nuts from traditionally recognized species such as P. pinea, P. sibirica, P. koraiensis, or P. gerardiana.