Renzo Bortolomeazzi

Daily consumption of a high-phenol extra-virgin olive oil reduces oxidative DNA damage in postmenopausal women

Extra-virgin olive oils (EVOO), high in phenolic compounds with antioxidant properties, could be partly responsible for the lower mortality and incidence of cancer and CVD in the Mediterranean region. The present study aims to measure oxidative DNA damage in healthy human subjects consuming olive oils with different concentrations of natural phenols. A randomised cross-over trial of high-phenol EVOO (high-EVOO; 592 mg total phenols/kg) v. low-phenol EVOO (low-EVOO; 147 mg/kg) was conducted in ten postmenopausal women in Florence. Subjects were asked to substitute all types of fat and oils habitually consumed with the study oil (50 g/d) for 8 weeks in each period. Oxidative DNA damage was measured by the comet assay in peripheral blood lymphocytes, collected at each visit during the study period. Urine samples over 24 h were collected to measure the excretion of the olive oil phenols. The average of the four measurements of oxidative DNA damage during treatment with high-EVOO was 30 % lower than the average during the low-EVOO treatment (P=0.02). Urinary excretion of hydroxytyrosol and its metabolite homovanillyl alcohol were significantly increased in subjects consuming high-EVOO. Despite the small sample size, the present study showed a reduction of DNA damage by consumption of an EVOO rich in phenols, particularly hydroxytyrosol.

Improvement of extraction procedure for biogenic amines in foods and their high-performance liquid chromatographic determination

A high-performance liquid chromatography method is described for the simultaneous determination of the biogenic amines tryptamine, 2-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine in cheese. The optimization of the procedure for the extraction of amines from the matrix is described. The separation of dansyl derivatives of the amines was achieved by reversed-phase liquid chromatography with gradient elution, followed by UV detection at 254 nm. The mobile phase was acetonitrile-0.01 M phosphate buffer (pH 7)-water. Under these conditions, rapid elution of the amines in less than 13 min was obtained. Validation of the method included calibration experiments, addition of standard amines for the determination of amine recoveries and repeatability tests.

Rapid mixed mode solid phase extraction method for the determination of acrylamide in roasted coffee by HPLC-MS/MS.

In this work, a rapid and reliable purification method based on a single mixed solid phase extraction (SPE) column, for the determination of acrylamide in roasted coffee by liquid chromatography-tandem mass spectrometry, was developed. Deuterium labelled d(3)-acrylamide was used as internal standard. Acrylamide was extracted by 10 mL of water and the extract purified by a single SPE column consisting of 0.5 g of an in-house prepared mixture of C18, strong cation (SCX) and anion exchange (SAX) sorbents in the ratio 2/1.5/1.5 (w/w/w). The amount of the three sorbents was optimised in order to eliminate the main interfering compounds present in coffee extracts, such as melanoidins, trigonelline, chlorogenic acids and caffeine. The SPE procedure was very simple and consisted of pushing 1 mL of an aqueous coffee extract through the SPE column followed by 1 mL of water which was collected for the analysis. The method was tested on six samples of roasted coffee of different composition and roasting level. The repeatability of the method, expressed as relative standard deviation (n=6), was lower than 5%. The recovery of acrylamide at three spiked levels ranged from 92% to 95%. The limits of detection (LOD) and quantitation (LOQ) were 5 and 16 μg kg(-1), respectively.

Composition of phenolic compounds and antioxidant activity of commercial aqueous smoke flavorings.

The antioxidant activity of 12 aqueous commercial smoke flavorings used in the food industry was determined by two methods: bleaching of the carotenoid crocin and scavenging of the DPPH radical. The reaction with the DPPH radical was evaluated by calculating the effective concentration (EC50) and the antiradical efficiency (AE). A gas chromatography-mass spectrometry method was, moreover, used for the determination of 2-methoxyphenols, 2,6-dimethoxyphenols, and dihydroxybenzenes. The methoxyphenols were extracted from the aqueous smoke by dichloromethane, and also the residue aqueous phase was analyzed to determine the more water-soluble dihydroxybenzenes. The recovery and the repeatability of the method are reported. The total phenolic concentrations of the smoke flavorings showed a wide range, from about 1000 to 25000 mg/kg. Considering the three classes of compounds, the concentrations were about 300-3000 mg/kg for the 2-methoxyphenols, 200-11000 mg/kg for the 2,6-dimethoxyphenols, and 140-10000 mg/kg for the dihydroxybenzenes. The range of the antioxidant activities of the smoke flavorings was wide, reflecting the wide range of the phenolic concentrations. Good correlations were obtained between the total phenolic concentration and the antioxidant activities determined by both the DPPH and crocin assays.

Oxisterol determination in selected coffees.

The main aim of green-coffee processing techniques, such as decqffeination and roasting, is always to maintain a very high level of quality in taste and flavor, the beverages most important cliaracteristics to consumers. Oxidative alterations of coffee lipids, which can occur in roasting, exert a very marked influence on these quality traits. Determining the extent of oxidation thus can provide an indication of the products potential shelf-life and reveal traces of any newly-formed oxidative products that might prove nutritionally unsafe. Yet, while much attention has recently been focused on certain by-products induced by cholesterol oxidation and their proven toxicity as risk factors in atherosclerosis and cancer, oxidated phytosterols have largely gone unnoticed, being considered along with β-sitosterol as not very dangerous in that neither is absorbed by the intestinal tract. The present study investigates the substances derived from phytosterol oxidation (oxisterols) in samples of regular and decaffeinated commercial coffees. The findings show that oxisterols were absent in some samples and that the traces of oxidate phytosterols detected in others were well below the threshold considered as toxicologically active.

Effect of vacuum roasting on acrylamide formation and reduction in coffee beans

Coffea arabica beans were roasted in an oven at 200 °C for increasing lengths of time under vacuum (i.e. 0.15 kPa). The samples were then analysed for colour, weight loss, acrylamide concentration and sensory properties. Data were compared with those obtained from coffee roasted at atmospheric pressure (i.e. conventional roasting), as well as at atmospheric pressure for 10 min followed by vacuum treatment (0.15 kPa; i.e. conventional-vacuum roasting). To compare the different treatments, weight loss, colour and acrylamide changes were expressed as a function of the thermal effect received by the coffee beans during the different roasting processes. Vacuum-processed coffee with medium roast degree had approximately 50% less acrylamide than its conventionally roasted counterpart. It was inferred that the low pressure generated inside the oven during the vacuum process exerted a stripping effect preventing acrylamide from being accumulated. Vacuum-processed coffee showed similar colour and sensory properties to conventionally roasted coffee.

Determination of polycyclic aromatic hydrocarbons in water and water-based alcoholic beverages

This paper proposes a simple HPLC method for the determination of polycyclic aromatic hydrocarbons (PAHs) in water, wine and beer. Samples were purified by PAH collection in solid-phase extraction (SPE) and analysed by reversed-phase HPLC (Supelcosil LC-PAH column from Supelco). For the beer sample, recoveries amounted to 28% for naphthalene and varied from 57% to 103% for the other PAHs; results are quantitative starting from fluoranthene (FI, the seventh component eluted). Almost all the beer and wine samples showed the presence of benzo(b)fluoranthene (BbF), benzo(k)fiuoranthene (BkF), benzo(a)pyrene (BaP), benz(ghi)perylene (BghiP) and indeno(1,2,3-cd)pyrene (IP), and in some cases there were traces of FI, benzo(a)anthracene (BaA) and dibenz(ah) anthracene (DBahA). Total contents of PAHs ranged from trace amounts to 0.72 ppb. Traces of BbF, BkF, BaP, BghiP and IP were also found in the wine samples.

Chromatographic determination of the position and configuration of isomers of methyl oleate hydroperoxides

Abstract Oxygen can react with organic substrates (RH) to yield hydroperoxides, which in many instances are the first products of oxidation to be analytically isolated. A study of the structure of hydroperoxides could elucidate the reaction mechanisms activated during the first steps of oxidation processes. The simplest structural model used in the study of oxidation mechanisms of fats is methyl oleate. In this work the structures of methyl oleate hydroperoxides (MOHPs) were determined by gas chromatography—ion-trap detector mass spectrometry (GC—ITD-MS) of the corresponding hydroxystearates (MSHs). The hydroperoxides were reduced to methyl hydroxyoctadecenoates (MOHs), which were separated into the cis and trans fractions by argentation thin-layer chromatography. By hydrogenation of the double bond the cis - and trans -MOHs were reduced to MSHs for GC—ITD-MS analysis. Methods to isolate and determine the positional isomers of MOHPs were tested. The analytical techniques used were preparative high-performance liquid chromatography and GC—ITD-MS, solid-phase extraction-GC—ITD-MS and direct GC—ITD-MS.

Thermal degradation of single 7-cholesteryl acetate hydroperoxide

SummaryThe thermal degradation products of the 7α- and 7β-cholesteryl 3β-acetate hydroperoxides obtained by thermal oxidation of cholesteryl 3β-acetate are reported and discussed. The hydroperoxides were degraded in both the vapour and condensed phases. The main components identified were 7-ketocholesteryl 3β-acetate, 7α- and 7β-hydroxycholesteryl 3β-acetates and the products generated from the loss of the hydroperoxide group and/or of acetic acid. Remarkable is the finding of the epoxy-hydroxy derivatives of cholesteryl acetate among the degradation products.

Rapid determination of cholesterol oxidation products in milk powder based products by reversed phase SPE and HPLC-APCI-MS/MS

A rapid and sensitive HPLC-APCI-MS/MS method for the determination of cholesterol oxidation products (COPs) in milk powder based foods is reported. The method consists in the direct saponification of the sample and purification of oxysterols by reversed phase C18-SPE followed by HPLC-MS/MS analysis. By this procedure, the extraction and enrichment of oxysterols are combined in a unique step, reducing sample manipulation and the possible formation of artifacts. LOD and LOQ were in the concentration ranges of 2-8ngg-1 and 8-30ngg-1, respectively. The precision (CV%) was in the range 10-36% in fresh samples with a total COPs amount from 212 to 645ngg-1 and 6-14% for an oxidized sample with a higher amount (3651ngg-1). The recovery ranged from 74±8% for 7-ketocholesterol to 101±12% for 7α-hydroxycholesterol at 200ngg-1 and from 82±2% for 7-ketocholesterol to 117±10% for 5α,6α-epoxycholesterol at 500ngg-1 spiked levels, respectively.