Anita Bufa

Volatile Composition of Macedonian and Hungarian Wines Assessed by GC/MS

The volatile composition of eight varietal wines Vranec, Merlot, Cabernet Sauvignon, Temjanika and Chardonnay from the Republic of Macedonia, and Portugieser, Kékfrankos and Tokaji Aszú from Hungary has been characterized by means of gas chromatography/mass spectrometry technique. The wine volatile compounds were extracted in dichloromethane, and the extracts were concentrated under nitrogen. Forty-four volatile compounds have been identified mainly using the NIST mass spectral library and by comparison with the available standards used for quantification as well. Differences between the wines were noted for a number of compounds, such as a higher concentration of 1-pentanol and 2-phenyl ethanol in the red wines. Monoterpenes, linalool and terpineol were detected only in the white wines, Chardonnay and Tokaji. Macedonian red wines were characterized by a higher level of alcohols, while the Hungarian wines contained a higher amount of esters, fatty acids and lactones. A statistical treatment including one-way ANOVA, followed by a Tukey’s test has been performed in order to ascertain possible significant differences between the wines studied, and principal component analysis to study the possible grouping of the wines.

Urinary Steroid Measurements in Some Endocrine and Psychiatric Diseases

In 1990, the worldwide accepted Shackleton method, which provides a possibility of determining the steroid metabolites from urine, was adopted in our laboratory. The procedure is very useful in the diagnosis of different endocrine diseases and in the recognition of dysfunction or absence of enzymes with an important role in steroid metabolism, and it gives possibility to control the treatment in patients with these diseases. Besides the proximate clinical application, the method gives a convenient tool to study the steroid background of these disorders, helping us understand the mechanism of their development. In the last few years, we have examined the steroid profile of patients with hair (androgen alopecia /AA/, effluvium /E/), psychiatric problems (major depression /MD/, eating disorders /EDS/, especially anorexia nervosa and bulimia) and osteoporosis (OP). In all of the examined hair loss diseases, the levels of main androgen metabolites were increased, and elevated 5alpha-reductase activity were found. We could observe the alteration of the activity of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) enzyme and marked gender differences in the changes of the steroid metabolism in patients with major depression (MD). In women with OP, the significantly decreased level of certain metabolites points to the role of testosterone, androstenedione and DHEA in postmenopausal bone loss in women. Our experiences contribute to the knowledge of the nature and steroid background of some endocrine and psychiatric diseases.

Urinary steroid profile in early pregnancy after in vitro fertilization

Objective. The aim of the study was to compare the levels of urinary steroid metabolites of patients with successful in vitro fertilization and patients who failed to achieve pregnancy. Design. Comparison of urinary steroid profiles prior to oocyte pick‐up and three weeks after embryo transfer. Setting. University hospital. Sample. Eleven women in the same age range with pregnancy after in vitro fertilization and eleven women who failed to achieve pregnancy. Methods. The standard “long” protocol was used for ovarian stimulation and intracytoplasmic sperm injection for assisted in vitro fertilization. The steroid metabolites in urine samples collected for 24 h were determined by gas chromatography–mass spectrometry. Main outcome measures. Steroid metabolite levels in urine samples determined in the early pregnancy period. Results. The levels of androsterone, etiocholanolone, pregnanediol, tetrahydro‐11‐dehydrocorticosterone and tetrahydro‐corticosterone were significantly higher (p < 0.05) in the urine of women with successful pregnancy three weeks after the embryo transfer, while the levels of tetrahydrocortisone, tetrahydrocortisol, allo‐tetrahydrocortisol and α‐cortolone became higher in the group of patients with unsuccessful pregnancy. Conclusions. The production of androgens, progesterone and corticoid steroid metabolites is altered in the early pregnancy period after in vitro fertilization.

Diagnostic relevance of urinary steroid profiles on ovarian granulosa cell tumors: two case reports

BackgroundGranulosa cell tumor of the ovary is the most frequent sex cord stromal tumor and represents 2 to 5% of all primary ovarian cancers. Ovarian granulosa cell tumor is a malignant tumor with slow progression and in some cases this tumor is hormonally active. The recurrence of granulosa cell tumor often happens after 5 years.Case presentationWe describe two cases of postmenopausal women with adult-type granulosa cell tumors of the ovary. Patient 1 is a 49-year-old European woman with a recurrent tumor; patient 2 is a 55-year-old European woman without recurrence of tumor. Urinary steroid profiles of patient 1 were monitored during a 5-year period starting from before an operation (13 samples). In patient 2, the urinary steroid profiles were monitored during a 3-year period starting from after an operation (six samples). The 24-hour urinary samples were examined and the urinary concentration of 20 androgen, progesterone, and corticoid metabolites was quantitatively determined by gas chromatography-mass spectrometry with selected ion-monitoring mode.ConclusionsBased on these cases a correlation could be observed between increased levels of the urinary steroids and the recurrence of ovarian granulosa cell tumor; therefore, we concluded that a urinary steroid profile could be a more effective method to follow-up such patients compared to the traditional serum hormones determinations supplemented with conventional tumor markers.

Advanced online mass spectrometry detection of proteins separated by capillary isoelectric focusing after sequential injection

Capillary isoelectric focusing hyphenated with mass spectrometry detection, following the sequential injection of the carrier ampholytes and the sample zone, is highly efficient for the characterization of proteins. The main advantage of the sequential injection protocol is that ampholytes, with pH ranges, which are not supposed to cover the isoelectric points of the sample components, can be used for separation. The method then allows online mass spectrometry detection of separated analytes either in the absence (substances that have left the pH gradient) or in the presence of low-level ampholytes (substances that are migrating within the pH gradient). The appearance of the substances within, or outside the pH gradient depends on, e.g., the composition of the ampholytes (broad or narrow pH range) or on the composition of electrolyte solutions. The experiments performed in coated capillaries (with polyvinyl alcohol or with polyacrylamide) show that the amount and the injection length of the ampholytes influence the length of the pH gradient formed in the capillary.

Altered urinary profiles of endogenous steroids in postmenopausal women with adenocarcinoma endometrii.

Objective. The potential role of androgen metabolism as co-factors in the development of carcinoma endometrii was investigated. Design. The urinary concentration of 23 androgen, progesterone and corticoid metabolites was quantitatively determined by gas chromatography–mass spectrometry with selected ion-monitoring. We obtained 24-h urine samples from 13 patients with adenocarcinoma endometrii and from 10 age-matched normal female subjects. In the course of the urinary steroid determination, we observed changes in the steroid profiles in the disease examined compared to the same age and same sex control group. Profiling urinary steroids has to give comprehensive information about the synthesis of steroids including the glandular and peripheral steroid metabolisms. Results. The concentrations of 16-hydroxy- dehydroepiandrosterone, pregnanediol and pregnenediol were not significantly different in the two groups. The concentrations of androsterone, etiocholanolone, 11β-hydroxy-androsterone, 11β-hydroxy-etiocholanolone, pregnanetriol, pregnenetriol, tetrahydrocortisone, tetrahydro-11-dehydrocorticosterone, tetrahydro-corticosterone, allo-tetrahydro-corticosterone, tetrahydrocortisol, allo-tetrahydrocortisol, α-cortolone, β-cortolone and α-cortol were significantly lower in the postmenopausal women with adenocarcinoma than in the controls. Conclusion. The changes in the concentrations of single metabolites point out the important role of steroid group, thus providing help in the recognition and treatment of diseased states.