R. Farré

Interlaboratory trial on the determination of the In Vitro iron dialysability from food

An in vitro method for the estimation of iron bioavailability was subjected to an interlaboratory trial. The method involved a simulated gastrointestinal digestion using pepsin for the gastric stage followed by pancreatin and bile salts for the intestinal stage. The proportion of iron diffused through a semipermeable membrane (molecular mass cut-off 10 kDa) was used to measure the iron dialysability. An interlaboratory trial between nine laboratories was conducted to evaluate the repeatability and reproducibility of the agreed method. The reproducibility of the method among the participating laboratories was 20-30% and depended on the content of dialysable iron. Several factors contributing to the variation in the in vitro dialysability among laboratories are discussed. The pH adjustment in the intestinal digestion was identified as one of the critical parameters. The present in vitro method was used to evaluate the iron dialysability from three meals. The dialysability data were in reasonable agreement with human absorption data. The usefulness of the in vitro dialysability method is discussed.

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.

High-performance liquid chromatographic determination of furfural compounds in infant formulas. Changes during heat treatment and storage.

Furfural contents in adapted and follow-up infant formulas were measured by RP-HPLC. The evolution of furfural compound contents during storage (a year at 20 and 37 degrees C) was studied. 2-Furylmethylketone and 5-methyl-2-furaldehyde were not detectable in analysed samples. The differences in the furfural compounds at point zero between both infant formulas has to be ascribed to the differences in protein and iron contents. An increase in free 5-hydroxymethyl-2-furfuraldehyde (HMF), 2-furaldehyde (F) and HMF+F contents was observed in all samples, although the differences were not statistically significant. The storage temperature affected the total HMF content and the storage time affected the total HMF and F contents.

Determination of ascorbic and dehydroascorbic acids in blood plasma and serum by liquid chromatography

A liquid-chromatography (LC) method with ultraviolet detection for measuring ascorbic (AA) and dehydroascorbic acid (DHA) in human blood and serum was studied. The method used an ODS reversed-phase column and cetyltrimethyl ammonium bromide as an ion-pairing agent. AA was measured before and after the reduction of DHA with dithiothreitol. The absence of interferences resulting from hemolysis products was verified and also the stability of the ascorbic acid in metaphosphoric acid extracts. The analytical parameters, linearity (1-80 micrograms/ml), accuracy (recovery, 96.7-100.7%) and precision (C.V. = 3.1%), show that the method is reliable and adequate for measuring the total vitamin C content in serum and plasma.

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.

Determination of liposoluble vitamins in cooked meals, milk and milk products by liquid chromatography

A method for the simultaneous determination of liposoluble vitamins in cooked meals was established. Saponification was performed with 50% (w/v) KOH at 80 degrees C, and ascorbic acid was added as antioxidant. The subsequent extraction was carried out with diethyl ether. This was followed by a liquid chromatographic separation on a reversed-phase C18 column with methanol-water (94:6, v/v as the mobile phase. Retinyl acetate was used as the internal standard. The analytical parameters linearity, detection limit (0.19 and 8.33 microg/100 g for retinol and alpha-tocopherol, respectively), precision of the method (RSD=5.24 and 6.99% for retinol and alpha-tocopherol, respectively) and recovery assays (95.6 and 96.5% for retinol and alpha-tocopherol, respectively) show that the method studied is useful for measuring these compounds in foods and cooked meals.

High-performance liquid chromatographic determination of Maillard compounds in store-brand and name-brand ultra-high-temperature-treated cows’ milk

Furosine and furfural products of the Maillard reaction are used as specific indicators of the effect of heating treatments on milk quality. Their contents were measured in representative samples of store- and name-brand ultra-high-temperature-treated milks using RP-HPLC with UV detection. Furosine contents ranged from 40.32 to 50.67 and from 65.48 to 310.58 mg/100 g protein in name- and store-brand milks, respectively. Of the furfurals, only hydroxymethylfurfural was detected. The free hydroxymethylfurfural contents of store-brand milks ranged from 0.22 to 1.70 mg/100 g protein. Total hydroxymethylfurfural contents ranged from 0.29 to 0.41 and from 0.72 to 2.21 mg/100 g protein, for name- and store-brands, respectively.

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.