María Jesús Lagarda

Lipid peroxidation and antioxidant enzyme activities in patients with type 1 diabetes mellitus.

To evaluate oxidative stress in type I diabetes mellitus, two antioxidant enzymes in erythrocytes, copper-zinc superoxide dismutase (SOD EC 1.15.1.1.) and seleno-dependent glutathione peroxidase (GSH-Px; EC 1.11.19), and two indexes of peroxidation in plasma, thiobarbituric acid reactive substances (TBARS) and organic hydroperoxides (OHP), were measured in 118 patients with insulin-dependent diabetes mellitus (IDDM), classified in accordance with the presence or absence of vascular complications and the degree of metabolic control established by the HbA1c level. Ninety healthy subjects made up the control group. According to our results, plasmatic TBARS and OHP concentrations are significantly higher in diabetics than in controls, and these differences are accentuated in diabetic people with vascular disorders. The GSH-Px activity was significantly reduced in diabetic patients with poor and medium metabolic control in relation to the control group, regardless of the existence or absence of vascular disorders. No differences in SOD activity between diabetic and control groups were found. A significant positive correlation between TBARS and HPO (r=0.683, p<0.001) was found in both the control and diabetic groups. Among the lipid parameters studied, there were only significantly positive correlations between TBARS and total cholesterol; TBARS and triglycerides; OHP and total cholesterol and OHP and triglycerides. Positive correlations between TBARS and HbA1c and between OHP and and HbA1c, and negative correlations between GSH-Px and HbA1c and between SOD and HbA1c were also found. The multiple regression analysis shows that TBARS and HPO correlate negatively with GSH-Px. There was no significant correlation with SOD.

Selenium, zinc and copper in plasma of patients with type 1 diabetes mellitus in different metabolic control states

The Studies of selenium (Se), zinc (Zn) and copper (Cu) levels in diabetic patients have led to contradictory findings as the possible relationship between the degree of diabetic control and the changes in mineral contents. In the present study the plasma Cu, Se, and Zn contents of diabetic patients and healthy people were measured and the relationship between these contents and diabetic metabolic control, as determined by glycosylated hemoglobin (HbA1c), was studied. The mean plasma Se content in diabetic patients was significantly lower than in controls (p < 0.01) and a negative correlation between the plasma contents of Se and HbA1c was found. No statistically significant differences in plasma Zn contents, either between patients with type 1 diabetes mellitus and control, were found. A statistically significant sex difference in plasma Cu contents was observed in the control population. In females, statistically significant differences were found in plasma Cu contents between the control subjects and the diabetic patients with medium or poor metabolic control, as well as between diabetic patients with good and poor metabolic control. In males, the only statistically significant differences were between the control subjects and diabetic patients with poor metabolic control. The correlation between plasma contents of Cu and HbA1c is not significant.

Bioaccessibility of tocopherols, carotenoids, and ascorbic acid from milk- and soy-based fruit beverages: influence of food matrix and processing.

A study was made of the effect of high-pressure processing (HPP) and thermal treatment (TT) on plant bioactive compounds (tocopherols, carotenoids, and ascorbic acid) in 12 fruit juice-milk beverages and of how the food matrix [whole milk (JW), skimmed milk (JS), and soy milk (JSy)] modulates their bioaccessibility (%). HPP (400 MPa/40 °C/5 min) produced a significant decrease in carotenoid and ascorbic acid bioaccessibility in all three beverages and maintained the bioaccessibility of tocopherols in JW and JS while decreasing it in JSy. TT (90 °C/30 s) produced a significant decrease in tocopherol and carotenoid bioaccessibility in all three beverages and increased the bioaccessibility of ascorbic acid. With regard to the food matrix, α-tocopherol and ascorbic acid bioaccessibility was greatest in JW beverages and lowest in JSy beverages, whereas no significant differences were found among the three beverages in terms of carotenoid bioaccessibility. HPP-treated samples showed higher tocopherol and carotenoid bioaccessibility than TT-treated samples, thus indicating that HPP combined with a milk matrix positively modulates the bioaccessibility of certain types of bioactive components of food, mainly those of a lipophilic nature.

Stability of Plant Sterols in Ingredients Used in Functional Foods

The content of plant sterol (PS) and their oxidation products (POPs) in eight ingredients used to enrich functional foods was studied. A gas chromatographic (GC) technique with mass-spectrometric detection was used for identification, while GC with a flame ionization detector (GC-FID) was used for quantification. β-Sitosterol was the most abundant phytosterol, and the main POPs found were derived from this compound (7α/β-hydroxysitosterol, 7-ketositosterol, and sitostanetriol). The total amount of POPs found in the ingredients ranged from 29.03 to 110.02 μg/100 g PS. The β-sitosterol oxidation rates ranged from 10 to 50 μg β-sitosterol oxides/100 g of β-sitosterol. In view of this low rate of oxidation in the ingredients tested, it can be concluded that the PS remain stable in these ingredients. Significant correlations (p < 0.01) were found between total oxysitosterols versus β-sitosterol contents (R(2) = 86.5%) and between total POPs and total PS (R(2) = 81.6%).

Environmental cadmium, lead and nickel contamination: possible relationship between soil and vegetable content

SummaryThe cadmium, lead and nickel content of soils of four agricultural areas exposed to different degrees of environmental pollution and vegetables grown there were measured by Atomic Absorption Spectrophotometry (AAS). In order to ascertain the possible relationship between the heavy metal content of soils (total and extractable) and of vegetables grown in them, the correlation between the two was calculated. The highest correlation values between metal content of soil and vegetables are these corresponding to nickel: the total metal content in soils and leaves-stems are linear (p<0.01). The extractable metal content of soils and leaves-stems are linear (p<0.1). Therefore, soil seems to be the main contributor to nickel content of plants (leaves-stems). In the case of cadmium a statistically significant linear correlation (p<0.01) was observed between total metal content in soil and in leaves-stems and a negative one between extractable metal content of soils and that of roots-bulbs. For lead a negative correlation was found between extractable metal content of soils and leaves-stems. In any case, the lead content of soils is not the main factor that influences the lead content of vegetables. Anthropogenic and environmental factors play a more important role than the lead in soils in the lead content of vegetables.

Methylmercury and inorganic mercury determination in fish by cold vapour generation atomic absorption spectrometry

Abstract Given that organic mercury is more dangerous than the inorganic form and that it is converted into methylmercury by biological methylation, we have studied and optimized a simple method for measuring both organic and inorganic mercury contents in fish, using a spectroscopic vapour generation technique, with a sequential reduction of the digested sample with stannous chloride and sodium tetrahydroborate. Prior to applying the method the sample was subjected to alkaline wet digestion. Due to the matrix interferences calibration curves with matrix addition were needed for mercury determinations. The analytical parameters of the method were: linearity from 10 to 200 ng of Hg in the reduction vessel; detection limit: 125 and 183 ng/g fresh sample for inorganic mercury and methylmercury, respectively; precision (RSD%): 9.8 and 10.1 for inorganic mercury and methylmercury, respectively; accuracy: reference material (Dorm-2-NRC-CNRC) for methylmercury; value found 4504±272 ng/g; certified value 4470±320 ng/g. The method offers the advantage of not requiring special equipment to measure inorganic and organic mercury simultaneously in a sample. To evaluate its usefulness it was applied to nine different types of fish and mussels.

Copper, iron and zinc determinations in human milk using FAAS with microwave digestion.

Abstract A method for determining copper, iron and zinc in human milk was optimized and validated. It includes microwave mineralization of the sample and measuring the elements by flame atomic absorption spectroscopy (FAAS). Only 2 ml of milk is needed, and the method is free of matrix interferences. The values obtained for the detection limits (0.07; 0.07; 0.11 μg/ml milk, for copper, iron and zinc) precision of the method, intra-assay (2.9; 5.2; 6.1%RSD for copper, iron and zinc) and accuracy, evaluated using recovery assays (98.8; 100.4; 95.9% for copper, iron and zinc) show that the method is useful for the purpose mentioned. Moreover, the method is rapid and simple, and the determinations are carried out by FAAS.

Whole blood selenium content in pregnant women

The selenium content in the blood of pregnant women in the Valencian Community, Spain was determined until the end of gestation in order to assess its evolution and detect possible differences in relation to the values corresponding to non-pregnant women of fertile age. A total of 158 blood samples were obtained from 137 pregnant women. Samples were classified as: (a) first; (b) second; and (c) third trimester. Selenium was determined by a flow injection hydride atomic absorption spectrometric method (Seronorm whole blood was used as a quality control check). The selenium values obtained for the three trimesters followed a Gaussian distribution. The intervals for mean found were: (a) 75.7-85.5 micrograms/l; (b) 72.6-81.4 micrograms/l; and (c) 69.9-77.5 micrograms/l. Although a tendency to decrease was observed, no statistical significant differences between the three trimesters of the gestational period were observed. When we compare the values obtained with the blood selenium content of women of fertile age in the Valencian community (87.8-98.8 micrograms/l) by using an ANOVA test, statistically significant differences are found between the selenium values of women of fertile age and those corresponding to pregnant women. The differences, however, are very small and can be partially explained by the level of hemodilution during pregnancy, however, reference values of blood selenium contents in pregnant women should be taken into account.

Calcium bioavailability in human milk, cow milk and infant formulas - comparison between dialysis and solubility methods

Abstract The percentages of total, soluble and dialysable calcium of human milk, cow milk and milk and soy based infant formulas were determined in order to detect possible differences in the calcium bioavailability of the samples. For this purpose an in vitro method was applied to these four calcium sources. The ranking of the analysed samples in terms of calcium bioavailability depends on the criteria applied. Calcium ranked dialysis percentage was: cow milk>human milk>soy based formula>milk based formulas. Calcium ranked solubility percentage was: human milk>cow milk>soy-based formula>milk-based formulas. Comparison of the results of the in vitro assay with the information available on in vivo calcium absorption showed that the total soluble calcium contents agree with the in vivo absorption values better than with calcium dialysis percentages.

Optimization of iron speciation (soluble, ferrous and ferric) in beans, chickpeas and lentils

Abstract A spectrophotometric method with bathophenanthroline for iron determination that makes it possible to differentiate between iron (II) and iron (III) in total soluble iron in legumes (beans, chickpeas and lentils) was optimized. Sample size, volumes of reducing agent and bathophenanthroline were selected. Matrix interferences made it necessary to apply the addition’s method. To check the quality of the method, linearity and precision (RSD%) were determined. A linear response between 0.1 and 1.8 μg Fe/ml in the assay and precision values ranging from 2.1 to 6 for instrumental precision, and from 1.6 to 1.7 and 2.7 to 9.1, for intra- and inter-day assays, respectively were obtained. The application of the method to legumes indicated: total soluble iron ranging from 0.52 (microwave cooked legumes) to 5.01 mg/100 g dry matter in raw beans. The percentage of soluble iron (II) with respect to total and soluble iron ranged from non detectable to 14.8% and from non detectable to 50%, respectively.