Diversity and Content of Carotenoids and Other Pigments in the Transition from the Green to the Red Stage of Haematococcus pluvialis Microalgae Identified by HPLC-DAD and LC-QTOF-MS.

H. pluvialis is a unicellular freshwater alga containing many bioactive compounds, especially carotenoids, which are the strongest antioxidants among the pigments. This study evaluates the composition and content of carotenoids and other pigments in both stages of algae life cycle, especially in the green vegetative stage, less studied in comparison to the red stage. To determine the composition and content of carotenoids, a combination of HPLC-DAD and LC-QTOF-MS was used. The content of carotenoids in the green vegetative stage was significantly lower than in the red vegetative stage. In the green vegetative stage, 16 different carotenoids and other pigments were identified. Among the total 8.86 mg g-1 DW of pigments, 5.24 mg g-1 DW or 59% of them were chlorophyll a with its derivatives, and 3.62 mg g-1 DW or 41% of them were free carotenoids. After the transition from the green to the red stage, the carotenoid composition was replaced by secondary carotenoids, astaxanthin and its esters, which predominated in the whole carotenoid composition. In addition to free astaxanthin, 12 astaxanthin monoesters, 6 diesters and 13 other carotenoids were determined. The majority of 37.86 mg g-1 DW pigments were monoesters. They represented 82% of all pigments, and their content was about 5 times higher than both, diesters (5.91 mg g-1 DW or 12% of all) and free carotenoids (2.4 mg g-1 DW or 6% of all). The results of the study contribute to the data on the overall pigment composition and content of H. pluvialis algae and provide the basis for further improvement of cultivation of the H. pluvialis algae.

Identification and Content of Astaxanthin and Its Esters from Microalgae Haematococcus pluvialis by HPLC-DAD and LC-QTOF-MS after Extraction with Various Solvents.

Haematococcus pluvialis, a unicellular green microalga that produces a secondary metabolite under stress conditions, bears one of the most potent antioxidants, namely xanthophyll astaxanthin. The aim of our study was to determine the content of astaxanthin and its esterified forms using three different solvents-methyl tert-butyl ether (MTBE), hexane isopropanol (HEX -IPA) and acetone (ACE)-and to identify them by using high performance liquid chromatography coupled with diode array detection and the quadrupole time-of-flight mass spectrometry (HPLC-DAD and LC-QTOF-MS) technique. We identified eleven astaxanthin monoesters, which accounted for 78.8% of the total astaxanthin pool, six astaxanthin diesters (20.5% of total), while free astaxanthin represented the smallest fraction (0.7%). Astaxanthin monoesters (C16:2, C16:1, C16:0), which were the major bioactive compounds in the H. pluvialis samples studied, ranged from 10.2 to 11.8 mg g-1 DW. Astaxanthin diesters (C18:4/C18:3, C18:1/C18:3) were detected in the range between 2.3 and 2.6 mg g-1 DW. All three solvents were found to be effective for extraction, but MTBE and hexane-isopropanol extracted the greatest amount of free bioactive astaxanthin. Furthermore, MTBE extracted more low-chain astaxanthin monoesters (C16), and hexane-isopropanol extracted more long-chain monoesters (C18 and above) and more diesters. We can conclude that MTBE is the solvent of choice for the extraction of monoesters and hexane-isopropanol for diesters.

Simultaneous determination of phthalates, their metabolites, alkylphenols and bisphenol A using GC-MS in urine of men with fertility problems.

A GC-MS method was successfully applied to measure simultaneously the concentrations of endocrine disrupting compounds (5 dialkyl phthalates, 9 phthalate monoesters, 3 alkylphenols and bisphenol A) in 136 male urine samples. In the present study the method was validated and concentrations of EDCs were determined. The results were compared with results from other studies. Correlations between endocrine disrupting compounds and also correlations of endocrine disrupting compounds with two semen quality parameters are presented and evaluated. Significant positive correlations were found between almost all the endocrine disrupting compounds. The parameter sum of DEHP (SUM DEHP) was positively correlated to all the endocrine disrupting compounds but negatively to two semen quality parameters. Negative correlations between the endocrine disrupting compounds and the semen quality parameters could indicate that endocrine disrupting compounds could cause reproductive problems by decreasing the semen count and quality. This research will have helped to evaluate human exposure to endocrine disrupting compounds.

Are urinary bisphenol A levels in men related to semen quality and embryo development after medically assisted reproduction?

OBJECTIVE: To evaluate whether urinary bisphenol A (BPA) levels in men adversely influence semen quality and embryo development after medically assisted reproduction. DESIGN: Prospective, cohort study. SETTING: University-based tertiary care center. PATIENT(S): A total of 149 couples undergoing their first or second IVF or intracytoplasmic sperm injection (ICSI) procedure. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Semen quality and embryo development parameters until the blastocyst stage after the IVF or ICSI procedure. RESULT(S): Bisphenol A was detected in 98% (n = 146) of the samples with 0.1 ng/mL limit of detection. The geometric mean BPA concentration was 1.55 ng/mL. After the adjustment for potential confounders using linear regression models, an increase of natural logarithm transformed urinary BPA concentration was associated with lower natural logarithm transformed sperm count (ß = -0.241, 95% confidence interval [CI] -0.470 to -0.012), natural logarithm transformed sperm concentration (ß = -0.219, 95% CI -0.436 to -0.003), and sperm vitality (ß = -2.660, 95% CI -4.991 to -0.329). The embryo development parameters from oocyte fertilization to the blastocyst formation stage were not affected by BPA exposure. CONCLUSION(S): Urinary BPA concentrations in male partners of subfertile couples may influence semen quality parameters, but do not affect embryo development up to the blastocyst stage after medically assisted reproduction.

Characterization of Slovenian wines using Multidimensional data analysis from simple enological descriptors.

Determination of the product's origin is one of the primary requirements when certifying a wine's authenticity. Significant research has described the possibilities of predicting a wine's origin using efficient methods of wine components' analyses connected with multivariate data analysis. The main goal of this study was to examine the discrimination ability of simple enological descriptors for the classification of Slovenian red and white wine samples according to their varieties and geographical origins. Another task was to investigate the inter-relations available among descriptors such as relative density, content of total acids, non-volatile acids and volatile acids, ash, reducing sugars, sugar-free extract, SO2, ethanol, pH, and an important additional variable - the sensorial quality of the wine, using correlation analysis, principal component analysis (PCA), and cluster analysis (CLU). 739 red and white wine samples were scanned on a Wine Scan FT 120, from wave numbers 926 cm(-1) to 5012 cm(-1). The applied methods of linear discriminant analysis (LDA), general discriminant analysis (GDA), and artificial neural networks (ANN), demonstrated their power for authentication purposes.