Jasminka Giacometti

Gas chromatographic determination of fatty acids contained in different lipid classes after their separation by solid-phase extraction

A quick and successful procedure is presented for the separation of polar lipids, monoacylglycerols (MAGs), diacylglycerols and triacylglycerols (TAGs) and for fatty acid determination in the above-mentioned lipid fractions by gas chromatographic analysis, which was acceptable for physiological and nutrition studies. In the analysis of edible oils and biological tissue samples, lipid classes were separated and purified by solid-phase extraction (SPE) using an aminopropylsilica column. Fatty acids in the sn-2 position in edible oil TAGs were determined after previous 1,3-specific lipase hydrolysis and separation of 2-MAGs by SPE using an aminopropylsilica column. A preliminary study of the solid-phase extraction separation of lipid classes with stock standard solutions using styrene-divinylbenzene-methacrylate copolymer (Nexus), octadecylsilica (C18) and aminopropylsilica (NH2) was carried out and it was shown that NH2 was the best sorbent for the above-mentioned purpose.

Determination of aliphatic alcohols, squalene,α-tocopherol and sterols in olive oils: direct method involving gaschromatography of the unsaponifiable fraction following silylation

In general, analyses for aliphatic alcohols, sterols and tocopherols in vegetable oils are performed separately. A simple and reliable procedure is presented for the quantification of the alkanols, squalene, alpha-tocopherol and sterols in olive oils by a direct method involving gas chromatographic (GC) analysis of the unsaponifiable fraction after silylation. The method eliminates the need for a preliminary thin-layer chromatographic (TLC) fractionation prior to GC. External standard calibration with reference substances was used for the quantification of squalene, alpha-tocopherol and sterols and internal standard calibration for the quantification of aliphatic alcohols. The analyte recovery and the repeatability of the quantitative results were evaluated and were acceptable for routine use.

Sex-specific differences of craniofacial traits in Croatia: The impact of environment in a small geographic area

Background: Craniometric variation in humans reflects different genetic and environmental influences. Long-term climatic adaptation is less likely to show an impact on size and shape variation in a small local area than at the global level. Aim: The aim of this work was to assess the contribution of the particular environmental factors to body height and craniofacial variability in a small geographic area of Croatia. Subjects and methods: A total of 632 subjects, aged 18–21, participated in the survey. Body height, head length, head breadth, head height, head circumference, cephalic index, morphological face height, face breadth, and facial index were analysed regarding geographic, climatic and dietary conditions in different regions of the country, and correlated with the specific climatic variables (cumulative multiyear sunshine duration, cumulative multiyear average precipitation, multiyear average air temperatures) and calcium concentrations in drinking water. Significant differences between groups classified according to geographic, climatic or dietary affiliation, and the impact of the environmental predictors on the variation in the investigated traits were assessed using multiple forward stepwise regression analyses. Results: Higher body height measures in both sexes were significantly correlated with Mediterranean diet type. Mediterranean diet type also contributed to higher head length and head circumference measures in females. Cephalic index values correlated to geographic regions in both sexes, showing an increase from southern to eastern Croatia. In the same direction, head length significantly decreased in males and head breadth increased in females. Mediterranean climate was associated with higher and narrower faces in females. The analysis of the particular climatic variables did not reveal a significant influence on body height in either sex. Concurrently, climatic features influenced all craniofacial traits in females and only head length and facial index in males. Mediterranean climate, characterized by higher average sunshine duration, higher average precipitation and higher average air temperatures, was associated with longer, higher and narrower skulls, higher head circumference, lower cephalic index, and higher and narrower faces (lower facial index). Calcium concentrations in drinking water did not correlate significantly with any dependent variable. Conclusion: A significant effect of environmental factors on body height and craniofacial variability was found in Croatian young adult population. This effect was more pronounced in females, revealing sex-specific craniofacial differentiation. However, the impact of environment was low and may explain only 1.0–7.32% variation of the investigated traits.

Determination of standard zinc values in the intact tissues of mice by ICP spectrometry.

The most commonly encountered difficulties for the quantitative measurement of zinc in biological samples are the limited sample amount, total and effective digestion of connective and fatty residues, and low zinc concentrations. These problems often lead to the determination of lower zinc values than actually present, so that the sample preparation, digestion, and analytical procedure deserve careful attention.In this short communication, a new method for microwave tissue disintegration is described. The authors have obtained consistent and reproducible results with tissue samples of 0.5 g or less.

Kinetic characterisation of enzymatic esterification in a solvent system: adsorptive control of water with molecular sieves

Abstract The kinetics of enzymatic esterification of glycerol with oleic acid, in equimolar ratio, catalyzed by immobilized Mucor miehei lipase in a solvent system in the presence of the molecular sieves was carried out at 37°C at different Lipozym and solvent ( n -hexane) concentrations and the molecular sieve contents were studied in a batch stirred-tank reactor (BSTR). The reactions were followed by the determination of reaction conversions during 45 h. The experimental data of enzymatic esterification of glycerol with oleic acid in a solvent system in the presence of molecular sieves showed minimal deviation from the calculated value in the irreversible second order kinetic model. On the basis of the experimental data, we found an empirical correlation between concentrations of Lipozym , concentrations of solvent ( n -hexane), contents of the molecular sieve and the reaction rate constant, k 1 .

Physico-chemical characteristics of olives and olive oil from two mono-cultivars during various ripening phases

The aim of this study was to find a correlation between the chemical and physical parameters in the pulp of the olive, in order to identify the optimum period during the 1998 harvesting period. For this reason various parameters have been monitored: the amount of oil in the pulp during the ripening phase obtained by solvent extraction with n-hexane, the acidity, the peroxide value, the composition of fatty acids and the saturated/unsaturated fatty acid ratio, the composition of the unsaponifiable fraction, the antioxidant capacity, the percentage of oil and the puncture test in two local Croatian olive cultivars. The chemical parameters change during the ripening, in order to control the oxidative changes of the cellular homeostasis of the whole lipophilic fraction that needs a stronger safeguard of the double bonds against the oxidative stress. The modification of the physical characteristics, observed with the puncture test, is related to the chemical changes which occur in the olive during the ripening period and the monitoring of these physical and chemical parameters can help to increase nutritional quality (as antioxidant capacity) and allow for an acceptable amount of oil.

Effect of Olive Oil- and Corn Oil- Enriched Diets on the Tissue Mineral Content in Mice

The mineral content (zinc, iron, magnesium, and calcium) in the liver, spleen, and thymus of male Balb/C mice was analyzed. Animals were fed, over 21 d, diets enriched with corn oil (FCO diet) or olive oil (FOO diet) (5% addition to standard pellet, w/w). Olive oil with predominant oleic acid (C18:1, n-9) had a quite different composition than corn oil, in which linoleic acid (C18:2, n-6) prevails. The zinc and magnesium tissue concentrations were not changed in either group. The calcium concentration in liver as well as the calcium concentration in spleen increased in mice fed both the FCO and FOO diets. Furthermore, mice fed both the FOO and FCO diets had increased spleen iron concentration. Mice fed the FCO diet had increased thymus calcium concentration compared to controls. The results show the effect of diets with unsaturated, particularly polyunsaturated fatty acids, on the calcium and iron concentration in some organs.

BanI polymorphism of cytosolic phospholipase A2 gene is associated with age at onset in male patients with schizophrenia and schizoaffective disorder

The enzymes phospholipases A2 are believed to be involved in the pathology of schizophrenia. We investigated allelic and genotype frequencies of PLA2G4A BanI polymorphism and the rs4375 in PLA2G6A in Croatian schizophrenic patients (n=81) and controls (n=182), using PCR/RFLP. Genotype and allelic frequencies of both loci, alone or in combination did not show significant difference (chi2-test). Allele-wise and genotype-wise meta-analyses of BanI polymorphism in case-control and family-based studies also revealed no significant association with schizophrenia. Multiple logistic regression analyses revealed statistically significant association between several items from PANSS general psychopathology scale and BanI polymorphism in PLA2G4A. BanI polymorphism further showed a significant impact on mean age of the onset of disease in males (betaA1=0.351, P=0.021; Spearmans rA1=0.391, P=0.010) indicating lower mean age at admission in homozygous A2A2 males.

The impact of PLA2G4A and PTGS2 gene polymorphisms, and red blood cell PUFAs deficit on niacin skin flush response in schizophrenia patients

We investigated the etiology of the attenuated niacin skin flush response in schizophrenia patients. Skin response to topical niacin of 0.1M, 0.01 M, 0.001 M, and 0.0001 M concentrations was rated using method of volumetric niacin response (VNR) and correlated to two functional A/G polymorphisms in genes: phospholipase A2 group IVA (BanI of the PLA2G4A), and rs689466 of the prostaglandin synthase-2 (PTGS2). We further tested the possible correlation between niacin response and fatty acid (FA) content of red blood cells (RBCs). We detected statistically significant but weak impact of both polymorphisms on niacin flush response in schizophrenia patients. The dosage of the G alleles of both polymorphisms was associated with higher VNR values, although each polymorphic variant accounted for only 1% of the overall flush response variability. Regarding FA content, both n-3 and n-6 polyunsaturated FAs (PUFAs) were significantly reduced in the patient group, but an association with niacin sensitivity was not detected.

Monitoring the esterification of sorbitol and fatty acids by gas chromatography

Abstract Lauric acid esters of sorbitol, 1,4-sorbitan and 1,4:3,6-isosorbide were synthesized in laboratory. Esterification was carried out at 160°C with a catalyst for sorbtol cyclization and esterification, viz., p-toluenesulfonic acid. Lipid classes and polyols were separated and analysed by GC as trimethylsilyl ethers and esters. The purpose of the study was to determine the suitability of GC analysis of these products as a means of following the progress of reactions. The results confirmed the possibility of monitoring lauric acid and sorbitol esterification via the GC determination of lauric acid concentration.