José Baptista

Nutritional value of selected macroalgae

Macroalgae are traditionally used in human and animal nutrition. Their protein and fiber content have been widely studied and differ according to the species, their geographic origin and their seasonal conditions. In addition to their value for human nutrition, seaweeds have multiple therapeutically applications (e.g., weight control, hypocholesterolemic, antioxidant and antitumor activities, others) and, in general, contribute and promote human health. In the archipelago of the Azores, the consumption of seaweeds is widespread and accepted as a common practice in some islands. This work is aimed at providing information on the protein and fiber content of the locally consumed species, to promote this regional food product that can be potentially profitable from the biotechnology and commercial perspective, and also benefit public health, particularly, taking into account the low level of marine pollution in the Azores archipelago. Protein and fiber content of eight seaweeds (Porphyra sp., Osmundea pinnatifida, Pterocladiella capillacea, Sphaerococcus coronopifolius, and Gelidium microdon, Rhodophyta; Cystoseira abies-marina and Fucus spiralis, Phaeophyta; Ulva compressa, Chlorophyta) were determined using the Kjeldahl method and the Weende method, respectively. The protein content ranged from 6.81 to 26.62 of dry weight for C. abies-marina and U. compressa, respectively. Fiber content was generally higher as compared with that in seaweeds from other origins and ranged from 33.82 to 63.88 for O. pinnatifida and F. spiralis, respectively.

Comparison of catechins and aromas among different green teas using HPLC/SPME-GC

Green tea polyphenols (GTP) from Camellia sinensis (L.) O. Kuntze are the major water soluble components in tea liquor. The GTP extraction yield was determined using different extraction times from 10 to 60 min at 70°C, and also at different temperatures from 50°C to 100°C, keeping the extraction time constant. The composition of the GTP (catechins) from samples of different origins was determined by RP–HPLC, measuring the absorbance at 280 nm. The calibration curves of the catechins were linear between 5 ng and 2 μg, and their C.V. values for the quantitation were less than 4.0% for four replicates using four different sets of each type. The total epicatechin derivatives (EPD) and epigallocatechin-3-gallate (EGCG) from Azorean green tea, accounted for 74.5 and 47.9% (w/w) of the total GTP, respectively, and were compared with that from tea samples of different origins. The EGCG:caffeine ratio in Azorean green tea was fourfold higher as compared with others, which suggested a good source of a chemopreventive agent. The aroma composition of different green tea samples was compared using the SPME/GC headspace methodology.

Comparison of polyphenols and aroma in red wines from Portuguese mainland versus Azores Islands

Abstract trans-Resveratrol (trans-3,5,4′.trihydroxystilbene), piceid (resveratrol 3-O-β-D-glucopyranoside), together with their isomers and quercetin were measured in red wines from cultivars grown in Portuguese mainland and Azores Islands. Crude hydroxystilbenes and quercetin were extracted by solid-phase extraction-C18 cartridges, and eluted with ethyl acetate. Recoveries of added trans-resveratrol and quercetin were between 90.2 to 101% (mean 96.8%) and between 95.2 and 97.6% (mean 96.2%), respectively. The precision measured by coefficients of variation (overall mean) ranged from 1.8 to 3.3%, and the calibration curves were linear with the mean correlation coefficients of 0.9997 and 0.9978 for trans-resveratrol and quercetin, respectively. The wine polyphenolic compounds were separated by reverse-phase C18 HPLC column and identified by UV fast scanning, retention times and HPLC/MS. The average content of total stilbenes was 4.75 and 6.33 mg/l, that of quercetin was 4.38 and 5.57 mg/l, and the trans/cis ratio was 8.32 and 10.89 for piceid and 5.2 and 4.09 for resveratrol in Portuguese mainland and Azorean wines, respectively. The Azorean wine (Basalto) was notable in having the highest trans-resveratrol concentration. The aroma profile of red wine samples from different regions were compared using SPME/GC-MS headspace methodology.

Methodology for a rapid and simultaneous determination of total cysteine, homocysteine, cysteinylglycine and glutathione in plasma by isocratic RP-HPLC.

Alterations in the plasma aminothiols levels can be considered as important biomarkers for the diagnosis and screening of several human disorders, namely cardiovascular diseases. We aimed to optimize a rapid, sensitive and accurate RP-HPLC methodology with fluorescence detection, for the simultaneous determination of the total concentrations of cysteine, homocysteine, cysteinylglycine and glutathione in blood plasma, as well as its application to the evaluation of those thiols levels in plasma of a group of Azorean subjects. Aminothiols were reduced with tri-n-butylphosphine and derivatized with a thiol-specific fluorogenic reagent ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate. The thiols adducts were separated by an isocratic elution on a Platinum EPS C18 analytical column (53mm×7mm I.D., 3μm) using a phosphate buffer containing 4% of acetonitrile as a mobile phase. Results indicated an excellent linearity for all the analytes over their respective concentration ranges with correlation coefficients (r(2)) ≥0.99. The LOD for the four plasma thiols was lower than 0.10μmol/L, while LOQ varied from 0.5 to 15μmol/L. For both intra- and inter-day precision, the RSD (%) values were lower than 1.9%, and the CV (%) values were found under 0.5%. The recovery ranged from 92% to 100% indicating a high degree of the methods accuracy. This method allows a simultaneous, complete analysis of the four plasma aminothiols and the internal standard in 6min. By reducing the total run time, a larger number of analysis can be performed daily.

Edible Azorean macroalgae as source of rich nutrients with impact on human health

Fucus spiralis, Porphyra sp. and Osmundea pinnatifida are macroalgae consumed as food in some of the Azorean Islands, but little information is available on their nutritional value. This paper presents information on the nutritional aspects in terms of proteins (9.71-24.82%), lipids (5.23-8.88%), soluble carbohydrates (17.59-25.37%), ash (22.31-38.55%), minerals (0.96-1.82 for Na/K ratios) and fat soluble vitamins A (1.20-1.41%) and E (4.86-51.14%) contents on dry weight basis. Fatty acid (SFA, MUFA, PUFA and n6/n3 and h/H ratios) and amino acid profiles, and antioxidant properties were also reported. All the three species contained measurable quantities of 9 of 10 essential amino acids (41.62-63.50% of the total amino acids). The regular consumption of these algae allied with the low level of seawater pollution around Azores Islands will improve human health conditions and will have a protective effect on the most common degenerative diseases.

Rapid, sensitive and simultaneous determination of ascorbic and uric acids in human plasma by ion-exclusion HPLC-UV.

OBJECTIVES Ascorbic (AA) and uric (UA) acids act as antioxidants and are capable to react with biologically relevant oxidants. We aimed to developed a simple, rapid, sensitive, and accurate ion-exclusion HPLC-UV methodology for the simultaneously determination of AA and UA in human plasma. METHODS Analytical pre-requisites, such as the use of heparin as an anticoagulant and meta-phosphoric acid as a stabilizer were added for accurate and reliable measurements. Chromatographic separation was achieved by an isocratic elution on a HEMA-BIO 1000 SB analytical column using a phosphate buffer, pH 2.4, as a mobile phase. RESULTS Results indicated an excellent linearity with correlation coefficients (r(2))≥0.999. The LOD of AA and UA was 1.02 and 1.42nmol/mL, respectively, while LOQ ranged from 0.306 to 0.426nmol/mL. A great repeatability for both antioxidants was found, where the CV (%) values for intra-day were lower than 1.8% and under 6.5% for the inter-day assay. The recovery of AA ranged from 92% to 96% and from 99% to 100% for UA. CONCLUSION This validated method allows the determination of both antioxidants within 10min, and is well suited to routine measurements and/or high-throughput clinical analysis. The methodology was applied to assess the antioxidant status of a group of Azorean subjects.

HPLC–UV–ESI-MS analysis of phenolic compounds and antioxidant properties of Hypericum undulatum shoot cultures and wild-growing plants

LC-UV and LC-MS analysis were used to study the phenolic composition of water extracts of Hypericum undulatum (HU) shoot cultures and wild-growing (WG) plants. Total phenolic content (TPC), determined using the Folin-Ciocalteu assay, and the antioxidant activity measured by two complementary methods were also performed for each sample. Mass spectrometry revealed several phenolics acids with quinic acid moieties, flavonols, mostly quercetin, luteolin and apigenin glycosides, flavan-3-ols (catechin and epicatechin) and the xanthonoid mangiferin. Differences in phenolic composition profile and TPC were found between the samples. The major phenolic in HU culture-growing (CG) samples is chlorogenic acid, followed by epicatechin, quercitrin and isoquercitrin. The WG plants presents hyperoside as the main phenolic, followed by isoquercitrin, chlorogenic acid and quercetin. The TPC and antioxidant activity were higher in samples from WG plants.

Comparison of the endemic Azorean Hypericum foliosum with other Hypericum species: antioxidant activity and phenolic profile

Water extracts of Hypericum foliosum (HF) from five Azorean Islands were analysed for their antioxidant activity and total phenolic contents. The results were compared with those from medicinal Hypericum species (H. perforatum, H. androsaemum and H. undulatum) and also with Azorean green tea. HF exhibited strong scavenging activity (87–91%) and moderate inhibition of linoleic acid oxidation (56–72%), and presented no significant difference to the other studied plants; in contrast, the synthetic antioxidant butylated hydroxyltoluene showed antioxidant activity values of 22% and 88%, respectively. The average value of phenolics in HF was higher than that in the other Hypericum species and lower than that in tea. Additionally, the phenolic profiles of the Hypericum species were compared by reverse phase-high performance liquid chromatography/ultraviolet (RP-HPLC/UV). The method presented permits the simultaneous determination of phenolic acids, flavonoids, hypericin and hyperforin within 55 min. The most similar profile was observed between HF and H. androsaemum. This study indicates that HF is a source of bioactive compounds with potential health benefits.

Fatty acid composition of selected macrophytes

The content of total lipids and the fatty acid (FA) profile were determined for eight macroalgae (Cystoseira abies-marina, Fucus spiralis, Chaetomorpha pachynema, Codium elisabethae, Porphyra sp., Osmundea pinnatifida, Pterocladiella capillacea and Sphaeroccoccus coronopifolius). Total lipids were extracted using a solvent mixture of methanol/chloroform (2/1, v/v) and further derivatised to FA methyl esters (FAME). The analyses of FAME samples were performed by gas chromatography coupled to a flame ionisation detector. The total lipid content ranged from 0.06 to 3.54 g (per 100 g). The most abundant saturated FA were palmitic (C16:0) and myristic (C14:0), while oleic (C18:1 n-9) was the dominant monounsaturated acid. All seaweeds contained linoleic FA (C18:2 n-6). The α-linolenic (C18:3 n-3) and eicosapentaenoic (20:5 n-3) acids were present only in Porphyra sp. (3.34% ± 0.13) and C. pachynema (0.47% ± 0.12), respectively. The n-6/n-3 and h/H ratios were low, suggesting a high nutritional value of the algae studied.

Assessment of molluscicidal activity of essential oils from five Azorean plants against Radix peregra (Müller, 1774)

The molluscicidal activity of essential oils from two endemic (Juniperus brevifolia; Laurus azorica) and three introduced (Hedychium gardnerianum; Pittosporum undulatum; Psidium cattleianum) Azorean plants against the snail Radix peregra was studied under laboratory conditions. Essential oils from leaves of H. gardnerianum, L. azorica and J.brevifolia presented promising molluscicidal activity on both adults and juveniles stages of R. peregra. The molluscicidal activity of these essential oils was found to be both time and concentration dependent. Lethal concentrations (LC(50)) varied between 15.4 (L. azorica) and 44.6 ppm (H. gardnerianum) for juveniles and from 45.3 (H. gardnerianum) to 54.6 ppm (J. brevifolia) for R. peregra adults. Ovicidal effect, calculated as percentage of egg hatching, at 100 ppm concentration, was observed in essential oils from P. undulatum flowers (4.2% of hatching) and leaves of H. gardnerianum (4.9%), L. azorica (7.4%) and J. brevifolia (17.7%). The present study is the first attempt to assess the molluscicidal potential of some Azorean plants essential oils against a Lymnaeidae snail. In fact, the H. gardnerianum, L. azorica and J. brevifolia can offer natural alternative tools for the control of R. peregra population, but more research is needed in order to determine the mode of action of these oils and determine the side effects on the ecosystem where this freshwater snail occurs.