High methoxyl pectin grafted onto gallic acid by one- and two-pot redox-pair procedures.

The aim of this research was to graft gallic acid (GA) onto high methoxyl pectin (HMP) through the redox-pair of ascorbic acid (Aa) and hydrogen peroxide (H2O2) with one- and two-pot procedures. The effectiveness of the both procedures and the chemical, physical and antioxidant properties of the obtained HMP-GA were evaluated. HMP-GAone-pot (23.3 ± 0.21 mg GA Equivalent (GAE)/g) and HMP-GAtwo-pot (32.3 ± 0.52 mg GAE/g) were best obtained at H2O2/Aa molar ratio-HMP/GA weight ratio of 9.0-0.5 and 16.0-0.5, respectively. The UV-Vis and FT-IR spectra and along with their derivative and thermal gravimetric analyses, revealed differences between HMP-GAone-pot and HMP-GAtwo-pot. The latter exhibited a greater antioxidant capacity than the former in single electron transfer (ET), hydrogen atom transfer (HAT), and ET-HAT mixed assays. The chemical differences can be attributed to side reactions that may have interfered with the grafting reaction. Consequently, HMP-GA, possessing unique antioxidant and prebiotic properties, can be synthesized through redox-pair procedures.

Structural, chemical and technofunctional properties pectin modification by green and novel intermediate frequency ultrasound procedure.

The impact of intermediate frequency ultrasound (IFUS, 582, 864 and 1144 kHz), mode of operation (continue and pulsed) and ascorbic acid (Aa) addition on the structural, chemical and technofunctional properties of commercial citrus high methoxyl-grade pectin (HMP) was investigated. The chemical dosimetry of IFUS, monitored by the triiodide formation rate (I3-), demonstrated that the pulsed ratio (1900 ms on/100 ms off) at the three frequencies was similar to that of continue mode but IFUS1144 kHz produced more acoustic streaming demonstrated by the height liquid measured using image analysis. In presence of Aa, HMP presented higher fragmentation than in its absence. IFUS did not give rise any changes in the main functional groups of the HMP. In general, a reduction in molecular weight was observed, being the presence of Aa the most influencing factor. Regarding monosaccharides, IFUS modified the structure of homogalacturonan and rhamnogalacturonan-I and increased of GalA contents of the HMP in presence of Aa at the above three frequencies. A reducing of the consistency index (k) and increasing of the flow index (n) of HMP were showed by IFUS frequency and Aa addition. The emulsifying activity and stability index were increased for HMP treated by IFUS in continue mode at all frequencies and in presence of Aa. The results presented in this research shown the effectiveness of IFUS as tool to modify pectin into different structures with different functionalities.

Impact of low-frequency ultrasound technology on physical, chemical and technological properties of cereals and pseudocereals.

Cereals (CE) and pseudocereals (PSCE) play a pivotal role in nourishing the human population. Low-frequency ultrasound (LFUS) modifies the structure of CE and PSCE macromolecules such as starch and proteins, often improving their technological, functional and bioactive properties. Hence, it is employed for enhancing the traditional processes utilized for the preparation of CE- and PSCE-based foods as well as for the upcycling of their by-products. We report recent advances in LFUS treatments for hydration, germination, extraction of bioactive compounds from by-products, and fortification of CEs and PSCE, including kinetic modelling and underlying action mechanisms. Meta-analyses of LFUS influence on compounds extraction and starch gelatinization are also presented. LFUS enhances hydration rate and time lag phase of CE and PSCE, essential for germination, extraction, fermentation and cooking. The germination is improved by increasing hydration, releasing promoters and eliminating inhibitors. Furthermore, LFUS boosts the extraction of phenolic compounds, polysaccharides and other food components; modifies starch structure, affecting pasting properties; causes partial denaturation of proteins, improving their interfacial properties and their peptides availability. Overall, LFUS has an outstanding potential to improve transformation processes and functionalities of CE and PSCE.

Hybrid high-intensity ultrasound and microwave treatment: A review on its effect on quality and bioactivity of foods.

With the growing of consumer's demand for products ready to eat that can be elaborated with greener technologies without affecting to their organoleptic characteristics, the application of ultrasound combined with microwaves has been widely studied on food preservation treatments (drying, frying), extraction of high-value added compounds and enzymatic hydrolysis of proteins. This review presents a complete picture of current knowledge on the ultrasound combined with microwaves including the mechanisms, influencing factors, advantages and drawbacks, emphasising in several synergistic effects observed in different processes of strong importance in the food industry. Recent research has shown that this hybrid technology could not only minimise the disadvantages of power US for drying and frying but also improve the product quality and the efficiency of both cooking processes by lowering the energy consumption. Regarding extraction, current studies have corroborated that the combined method presents higher yields in less time, in comparison with those in the respective ultrasound and microwave separately. Additionally, recent results have indicated that the bioactive compounds extracted by this combined technology exhibit promising antitumor activities as well as antioxidant and hepatoprotective effects. Remarkably, this hybrid technology has been shown as a good pre-treatment since the structural changes that are produced in the molecules facilitate the subsequent action of enzymes. However, the combination of these techniques still requires a proper design to develop and optimized conditions are required to make a scale process, and it may lead to a major step concerning a sustainable development and utilization of bioactive compounds from natural products in real life.

Design of polyphenol-rich diets in clinical trials: A systematic review.

Most randomized clinical trials of polyphenols focus on individual foods. Nevertheless, due to their presence in many foods and in order to reflect a real situation, clinical trials based on polyphenol-rich diets are particularly important. This systematic review explores the characteristics of the polyphenol-rich diets used in intervention studies. The bibliography search for English-language scientific papers was performed in the Elsevier Scopus Database and PUBMED in March 2020, and focused on intervention studies with whole polyphenol-rich diets, establishing several exclusion criteria. In studies fulfilling the requirements, information on the design of the polyphenol-rich diet and associated polyphenol intake was extracted and compared. A total of 5 studies were selected. Among them, substantial differences were found in the design of the polyphenol-rich diets, regarding specific instructions and concerning the foods provided. Similarly, although a median daily polyphenol intake of 2,564 mg/day (17,945 mg/week) was obtained from the studies, which corresponds to a nutritional dose, intake values varied widely both for total polyphenols (the difference between studies reached threefold), and for individual polyphenol intake (for hydroxycinnamic acids, a tenfold difference was found between percentile 25 and percentile 75 values). These differences made the comparison of results difficult and may affected the observed health effects. Thus, despite the relevance of studying polyphenol-rich diets as a whole, this systematic review found substantial differences between the studies performed, making direct comparisons difficult.

Physicochemical and functional properties of isolated starch and their correlation with flour from the Andean Peruvian quinoa varieties.

Quinoa has been recognized as a complete food due to its balanced nutritional composition. Quinoa flour is used as an ingredient to improve the nutritional and functional characteristics of cereal-based foods. The physicochemical and functional (thermal and pasting) properties of flours and isolated starches of three Andean Peruvian quinoa varieties (Blanca de Hualhuas, BH; Rosada de Huancayo, RHY and Pasankalla, PK) were studied and the correlation among them properties were evaluated in order to explore their possible uses as a food ingredient. Proximal chemical composition of flour and isolated starches from quinoa varieties showed differences. Isolated starches from quinoa varieties showed a XRD Type A crystallinity patterns with polygonal shapes, small size, higher crystallinity degree and lower amylose content (<15%). The thermal (gelatinization temperatures and enthalpies) and pasting (temperature and time of gelatinization and viscosities) properties of flours and isolated starches showed differences and the principal component analysis demonstrated that those properties are significantly correlated to the starch and fat content. Based on the differences found among physicochemical and functional properties, isolated starch and flour of BH, RHY and PK quinoa varieties have potential as food ingredient for several cereal-based products.

A green analytical assay for the quantitation of the total saponins in quinoa (Chenopodium quinoa Willd.) based on macro lens-coupled smartphone.

The aim of this study was to develop and validate a microtiter macro lens-coupled smartphone (MCS) assay for the quantitation of the total saponins in quinoa based on foam measurement. The 96-well micro plate with a black bottom and an inclination angle = 12.102° of macro lens-coupled smartphone allows to acquire images with a high resolution. The foam stability, a critical aspect for the MCS assay, was significantly improved by the inclusion of a chelating agent (EDTA 50 mmol L-1) and bovine serum albumin (0.5 mg mL-1). The MCS assay was linear within the range of 9.039-180.773 × 10-4 mg mL-1 of saponin (R2 = 0.9929), using the integrated density/area as a foam measurement (Y) and the logarithm of saponin concentration (X) (Y = 372.1 + 104.2LogX). The MCS assay was 50-folds more sensitive than afrosimetric assay -AA- with LOD = 3.168 × 10-4 mg mL-1 and LOQ = 4.784 × 10-4 mg mL-1. MCS assay was more reproducible (relative standard deviation (RSD) = 0.632-9.646%) than AA (RSD = 3.44-44.04%). The correlation analysis, Bland-Altman analysis and Passing-Bablok regression showed good agreement between total saponin content in quinoa as measured by the MCS assay and AA. Based on the green analytical procedure index, MCS assay can be considered as a green procedure.

Microencapsulated betacyanin from colored organic quinoa (Chenopodium quinoa Willd.): optimization, physicochemical characterization and accelerated storage stability.

BACKGROUND: Betalains are presently gaining popularity as pigments for use as natural colorants and/or bioactive compounds in functional foods. Quinoa (Chenopodium quinoa Willd.) has been recognized as an extremely nutritious grain and has recently been found to be a novel and good betalain source. Microencapsulation has been studied as a protected-delivery procedure to stabilize betalains. There are no studies about microencapsulation of betacyanins extracted from quinoa using spray-drying technology. RESULTS: Optimal microencapsulation was obtained at a drying temperature of 165 °C, a rotameter air flow rate of 47 mm (940 L h-1 ) and 10% w/w maltodextrin, which produced good encapsulation yield (58.1%) and efficiency (100%). Optimized maltodextrin-betacyanin microcapsules (diameter 4.4 µm) have low moisture (1.64 ± 0.08%) and water activity (0.127 ± 0.006), a betacyanin content of 0.1995 ± 0.0017 g kg-1 and saponin content <0.080 mg kg-1 . The oxygen consumption rate by betacyanin was -4.373 × 10-5 bar min-1 at 80 °C and -6.67 × 10-5 bar min-1 at 90 °C, which was accompanied by fading of the color. CONCLUSION: Microencapsulated betacyanin was optimized by response surface methodology, and its stability was measured under accelerated conditions by oxygen consumption. Microencapsulations contain betacyanin and low saponin concentration, which might confer unique health-promoting properties. © 2018 Society of Chemical Industry.

Role of fetal nutrient restriction and postnatal catch-up growth on structural and mechanical alterations of rat aorta.

KEY POINTS: Intrauterine growth restriction (IUGR), induced by maternal undernutrition, leads to impaired aortic development. This is followed by hypertrophic remodelling associated with accelerated growth during lactation. Fetal nutrient restriction is associated with increased aortic compliance at birth and at weaning, but not in adult animals. This mechanical alteration may be related to a decreased perinatal collagen deposition. Aortic elastin scaffolds purified from young male and female IUGR animals also exhibit increased compliance, only maintained in adult IUGR females. These mechanical alterations may be related to differences in elastin deposition and remodelling. Fetal undernutrition induces similar aortic structural and mechanical alterations in young male and female rats. Our data argue against an early mechanical cause for the sex differences in hypertension development induced by maternal undernutrition. However, the larger compliance of elastin in adult IUGR females may contribute to the maintenance of a normal blood pressure level. ABSTRACT: Fetal undernutrition programmes hypertension development, males being more susceptible. Deficient fetal elastogenesis and vascular growth is a possible mechanism. We investigated the role of aortic mechanical alterations in a rat model of hypertension programming, evaluating changes at birth, weaning and adulthood. Dams were fed ad libitum (Control) or 50% of control intake during the second half of gestation (maternal undernutrition, MUN). Offspring aged 3 days, 21 days and 6 months were studied. Blood pressure was evaluated in vivo. In the thoracic aorta we assessed gross structure, mechanical properties (intact and purified elastin), collagen and elastin content and internal elastic lamina (IEL) organization. Only adult MUN males developed hypertension (systolic blood pressure: MUNmales  = 176.6 ± 5.6 mmHg; Controlmales  = 136.1 ± 4.9 mmHg). At birth MUN rats were lighter, with smaller aortic cross-sectional area (MUNmales  = (1.51 ± 0.08) × 105  µm2 , Controlmales  = (2.8 ± 0.04) × 105  µm2 ); during lactation MUN males and females exhibited catch-up growth and aortic hypertrophy (MUNmales  = (14.5 ± 0.5) × 105  µm2 , Controlmales  = (10.4 ± 0.9) × 105  µm2 ), maintained until adulthood. MUN aortas were more compliant until weaning (functional stiffness: MUNmales  = 1.0 ± 0.04; Controlmales  = 1.3 ± 0.03), containing less collagen with larger IEL fenestrae, returning to normal in adulthood. Purified elastin from young MUN offspring was more compliant in both sexes; only MUN adult females maintained larger elastin compliance (slope: MUNfemales  = 24.1 ± 1.9; Controlfemales  = 33.3 ± 2.8). Fetal undernutrition induces deficient aortic development followed by hypertrophic remodelling and larger aortic compliance in the perinatal period, with similar alterations in collagen and elastin in both sexes. The observed alterations argue against an initial mechanical cause for sex differences in hypertension development. However, the maintenance of high elastin compliance in adult females might protect them against blood pressure rise.

A novel pyrogallol red-based assay to assess catalase activity: Optimization by response surface methodology.

Pyrogallol red (PGR) was identified as a novel optical probe for the detection of hydrogen peroxide (H2O2) based on horseradish peroxidase (HRP)-catalyzed oxidation. Response surface methodology (RSM) was applied as a tool to optimize the concentrations of PGR (100µmolL-1), HRP (1UmL-1) and H2O2 (250µmolL-1) and used to develop a sensitive PGR-based catalase (CAT) activity assay (PGR-CAT assay). N-ethylmaleimide -NEM- (102mmolL-1) was used to avoid interference produced by thiol groups while protecting CAT activity. Incubation time (30min) for samples or CAT used as standard and H2O2 as well as signal stability (stable between 5 and 60min) were also evaluated. PGR-CAT assay was linear within the range of 0-4UmL-1 (R2=0.993) and very sensitive with limits of detection (LOD) of 0.005UmL-1 and quantitation (LOQ) of 0.01UmL-1. PGR-CAT assay showed an adequate intra-day RSD=0.6-9.5% and inter-day RSD=2.4-8.9%. Bland-Altman analysis and Passing-Bablok and Pearson correlation analysis showed good agreement between CAT activity as measured by the PRG-CAT assay and the Amplex Red assay. The PGR-CAT assay is more sensitive than all the other colorimetric assays reported, particularly the Amplex Red assay, and the cost of PGR is a small fraction (about 1/1000) of that of an Amplex Red probe, so it can be expected to find wide use among scientists studying CAT activity in biological samples.

Maternal plasma antioxidant status in the first trimester of pregnancy and development of obstetric complications.

INTRODUCTION: Oxidative stress is present in pregnancy complications. However, it is unknown if early maternal antioxidant status could influence later development of complications. The use of assisted reproduction techniques (ART) is rising due to the delay of first pregnancy and there is scarce information on its influence on oxidative balance. OBJECTIVE: To assess the possible relationship between maternal plasma antioxidant status in first trimester of gestation with later development of pregnancy complications, evaluating the influence of ART and nutrition. METHODS: Plasma from 98 healthy pregnant women was obtained at week 10, nutrition questionnaires filled and women were followed until delivery. We evaluated biomarkers of oxidative damage (carbonyls, malondialdehyde-MDA), antioxidants (thiols, reduced glutathione, phenolic compounds, catalase and superoxide dismutase activities) by spectrophotometry/fluorimetry and melatonin (ELISA). Antioxidant status score (Antiox-S) was calculated as the computation of antioxidants. Diet-antioxidants relationship was evaluated through multiple correspondence analysis. RESULTS: Melatonin and carbonyls exhibited a negative correlation. No difference in oxidative damage was found between groups, but Antiox-S was significantly lower in women who developed complications. No differences in oxidative damage or Antiox-S were found between ART and no-ART pregnancies. High consumption of foods of vegetable origin cluster with high plasma levels of phenolic compounds and with high Antiox-S. CONCLUSIONS: In early normal gestation, low plasma antioxidant status, assessed through a global score, associates with later development of pregnancy complications. Larger population studies could help to determine the value of Antiox-S as predictive tool and the relevance of nutrition on maternal antioxidant status.

Prediction of fermentation index of cocoa beans (Theobroma cacao L.) based on color measurement and artificial neural networks.

Several procedures are currently used to assess fermentation index (FI) of cocoa beans (Theobroma cacao L.) for quality control. However, all of them present several drawbacks. The aim of the present work was to develop and validate a simple image based quantitative procedure, using color measurement and artificial neural network (ANNs). ANN models based on color measurements were tested to predict fermentation index (FI) of fermented cocoa beans. The RGB values were measured from surface and center region of fermented beans in images obtained by camera and desktop scanner. The FI was defined as the ratio of total free amino acids in fermented versus non-fermented samples. The ANN model that included RGB color measurement of fermented cocoa surface and R/G ratio in cocoa bean of alkaline extracts was able to predict FI with no statistical difference compared with the experimental values. Performance of the ANN model was evaluated by the coefficient of determination, Bland-Altman plot and Passing-Bablok regression analyses. Moreover, in fermented beans, total sugar content and titratable acidity showed a similar pattern to the total free amino acid predicted through the color based ANN model. The results of the present work demonstrate that the proposed ANN model can be adopted as a low-cost and in situ procedure to predict FI in fermented cocoa beans through apps developed for mobile device.

Short-term esmolol attenuates remodeling of the thoracic aorta in hypertensive rats by decreasing concentrations of ADMA down-regulated by oxidative stress.

Esmolol produces early regression of left ventricular hypertrophy and improves coronary artery remodeling, although the impact of short-term treatment with this beta-blocker on remodeling in large arteries has not yet been studied. We hypothesized that even a short (48h) course of esmolol might alter remodeling of the aorta in the spontaneously hypertensive rat (SHR). Fourteen-month-old male SHRs were treated intravenously with vehicle (SHR, n=8) or esmolol (SHR-E, n=8) (300µg/kg/min). Age-matched, vehicle-treated male Wistar-Kyoto rats (WKY, n=8) served as controls. After 48h, we studied the structure, volume density of elastic fibers, and passive mechanical properties of the aorta. Determination of asymmetrical dimethylarginine concentrations and total protein carbonyls in the aorta were analyzed. Esmolol significantly attenuated abnormal aortic wall thickness, cross-sectional area, wall-to-lumen ratio, volume density of elastic fibers, and wall stiffness. The protective effect of esmolol could be related to a decrease in asymmetrical dimethylarginine levels after down-regulation by oxidative stress. These findings could play a key role in the selection of antihypertensive therapy in patients with hypertension and aortic remodeling.

A novel high-throughput image based rapid Folin-Ciocalteau assay for assessment of reducing capacity in foods.

The aim of the presented work was to develop and validate a novel high-throughput rapid Folin-Ciocalteau assay for the quantification of reducing capacity of foods based on image scanner (Image-F-C assay). The original rapid F-C assay using a 96-well plate was improved by adding a neutralization step that stabilizes the formed color, enabling image acquisition using a flatbed scanner. Although the scanner has been already used in other analytical applications, no analysis has been reported regarding the effect of the scanner model, the plate orientation or the reaction volume. In the present study, we establish that the mentioned parameters do affect the linearity and precision of image based Folin-Ciocalteau assay, and provide the optimal scanning conditions for the analyzed scanner models. Euclidean distance calculated from R (Red), G (Green) and B (Blue) values was chosen, based on linearity and sensitivity, in order to quantify the reducing capacity. An in-house program using free ImageJ macro language was written to calculate automatically the RGB values of each well. The Image-F-C assay is linear within the range of 0-20 mg L(-1) of gallic acid (R(2)≥0.9939). We compared reducing capacity values from real samples quantified by the image F-C assay and by a microplate reader and an inter-day relative standard error<8% was observed. Bland-Altman and correlation analyzes showed that there were no significant differences between the two methods.

Fetal undernutrition is associated with perinatal sex-dependent alterations in oxidative status.

Intrauterine growth retardation predisposes to hypertension development, known as fetal programming. Females are less susceptible, which has been mainly attributed to estrogen influence. We hypothesize that perinatal differences in oxidative status might also contribute. We studied 21-day-old (prepuberal) and 6-month-old male and female offspring from rats fed ad libitum during gestation (Control) or with 50% of Control daily intake from day 10 to delivery (maternal undernutrition, MUN). We assessed in vivo blood pressure and the following plasma biomarkers of oxidative status: protein carbonyls, thiols, reduced glutathione (GSH), total antioxidant capacity, superoxide anion scavenging activity (SOSA) and catalase activities; we calculated a global score (oxy-score) from them. Estradiol and melatonin concentration was measured in young rats. Prepuberal MUN males were normotensive but already exhibited increased carbonyls and lower thiols, GSH, SOSA and melatonin; oxy-score was significantly lower compared to Control males. Prepuberal MUN females only exhibited reduced SOSA compared to Control females. Adult rats from all experimental groups showed a significant increase in carbonyls and a decrease in antioxidants compared to prepuberal rats; oxy-score was negative in adult rats suggesting the development of a prooxidative status as rat age. Adult MUN males were hypertensive and exhibited the highest increase in carbonyls despite similar or even higher antioxidant levels compared to Controls. Adult MUN females remained normotensive and did not exhibit differences in any of the biomarkers compared to Controls. The better global antioxidant status developed by MUN females during perinatal life could contribute to their protection against hypertension programming.

A novel, micro, rapid and direct assay to assess total antioxidant capacity of solid foods.

A novel, micro, rapid and direct procedure to measure the total antioxidant capacity of solid foods using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) (mR-QUENCHER-DPPH) was developed and validated. The mR-QUENCHER-DPPH assay was performed in semi-aqueous medium (methanol-Tris buffer) using very small sample amounts (below 3.6 µg), as estimated by a Bradford reagent-based chemical predictor, and it was completed in 10 min at room temperature. The total antioxidant capacity (TAC) of solid foods was expressed as scavenging capacity index (SCI, mmol DPPH scavenged per kg sample), a theoretical and stoichiometric parameter deduced in this study. SCI values measured by mR-QUENCHER-DPPH assay for cereals cous-cous (7.20±0.35), amaranth (7.99±0.35) and buckwheat (194.2±6.72); Goji fruit (91.27±3.98); lotus root (2402±168); and spices turmeric (3767±355), ginger (2493±283), and cinnamon (10461±2133) were further validated using Folin-Ciocalteau assay. Bland-Altman analysis showed that there were not statistically significant differences in TAC values as measured by both assays. In the same way, TAC values measured by mR-QUENCHER-DPPH were correlated with free (r=0.8088, P=0.0151), bound (r=0.9668, P<0.0001) and total (r=0.9067, P=0.0019) reducing capacity of extracts from solid foods as assessed by Folin-Ciocalteau assay. The mR-QUENCHER-DPPH assay allows to measure TAC values using micro-gram amounts in solid food samples with a wide content range of antioxidants (low, high and very high), and omitting the time-consuming dilution cellulose-step commonly employed in the traditional QUENCHER procedures.

Physical features, phenolic compounds, betalains and total antioxidant capacity of coloured quinoa seeds (Chenopodium quinoa Willd.) from Peruvian Altiplano.

Physical features, bioactive compounds and total antioxidant capacity (TAC) of coloured quinoa varieties (Chenopodium quinoa Willd.) from Peruvian Altiplano were studied. Quinoa seeds did not show a pure red colour, but a mixture which corresponded to different fractal colour values (51.0-71.8), and they varied from small to large size. Regarding bioactive compounds, total phenolic (1.23-3.24mg gallic acid equivalents/g) and flavonol contents (0.47-2.55mg quercetin equivalents/g) were highly correlated (r=0.910). Betalains content (0.15-6.10mg/100g) was correlated with L colour parameter (r=-0.569), total phenolics (r=0.703) and flavonols content (r=0.718). Ratio of betaxanthins to betacyanins (0.0-1.41) was negatively correlated with L value (r=-0.744). Whereas, high TAC values (119.8-335.9mmol Trolox equivalents/kg) were negatively correlated with L value (r=-0.779), but positively with betalains (r=0.730), as well as with free (r=0.639), bound (r=0.558) and total phenolic compounds (r=0.676). Unexploited coloured quinoa seeds are proposed as a valuable natural source of phenolics and betalains with high antioxidant capacity.

Improved evaporative light scattering detection for carbohydrate analysis.

Optimization and validation of evaporative light scattering detector (ELSD), aided by response surface methodology (RSM), has been developed for the liquid chromatography analysis of a wide molecular weight (MW) range of carbohydrates, including polysaccharides and oligosaccharides. Optimal experimental parameters for the ELSD detection were: 88.8°C evaporator temperature, 77.9°C nebulizer temperature and 1.1 standard litres per minute nitrogen flow rate. Optimal ELSD detection, used together with high performance size exclusion chromatography (HPSEC) of carbohydrates, gave a linear range from 250 to 1000 mg L(-1) (R(2)>0.998), with limits of detection and quantitation of 4.83-11.67 and 16.11-38.91 mg L(-1), respectively. Relative standard deviation was lower than 1.8% for intra-day and inter-day repeatability for apple pectin, inulin, verbascose, stachyose and raffinose. Recovery ranged from 103.7% to 118.3% for fructo-oligosaccharides, α-galacto-oligosaccharides and disaccharides. Optimized and validated ELSD detection is proposed for the analysis of high- to low-MW carbohydrates with high sensitivity, precision and accuracy.

Development of albuminuria and enhancement of oxidative stress during chronic renin-angiotensin system suppression.

OBJECTIVE: Albuminuria has been recently described in hypertensive patients under chronic renin-angiotensin system (RAS) suppression. We investigated whether this fact could be related to an increase in oxidative stress. METHODS: We examined normoalbuminuric and albuminuric patients in stage 2 chronic kidney disease, both with more than 2 years of RAS blockade. The relationship between albuminuria and circulating biomarkers for both oxidative damage, that is carbonyl and malondialdehyde, as well as antioxidant defense, that is reduced glutathione, thiol groups, uric acid, bilirubin, or catalase, and superoxide scavenging activity, was assessed. RESULTS: We found that only patients with albuminuria showed an important increase in carbonyls (P < 0.001) and malondialdehyde (P < 0.05) compared to normoalbuminuric patients. This increase in oxidative damage was also accompanied by a rise in catalase activity (P < 0.05) and low-molecular-weight antioxidants only when they were measured as total antioxidant capacity (P < 0.01). In order to establish the specific oxidative status of each group, new indexes of oxidative damage and antioxidant defense were calculated with all these markers following a mathematical and statistical approach. Although both pro-oxidant and antioxidant indexes were significantly increased in patients with albuminuria, only the oxidative damage index positively correlated with the increase of albumin/creatinine ratio (P = 0.0024). CONCLUSIONS: We conclude that albuminuria is accompanied by an amplified oxidative damage in patients in early stages of chronic kidney disease. These results indicate that chronic RAS protection must be directed to avoid development of albuminuria and oxidative damage.