The Development of an Extraction Method for Simultaneously Analyzing Fatty Acids in Macroalgae Using SPE with Derivatization for LC-MS/MS.

Fatty acid analysis is an essential step in evaluating the potential of macroalgae for biodiesel production. An extraction method was developed to simultaneously analyze up to five types of biodiesel-fuel-related fatty acids (myristic acid, palmitic acid, cis-palmitvaccenic acid, stearic acid, and oleic acid) in macroalgae using liquid chromatography and tandem mass spectrometry (LC-MS/MS). Lypophilization and solid-phase extraction (SPE) techniques were applied to improve the extraction efficiency and effectively purify samples. The optimal conditions for SPE were set by comparing the recoveries according to the various solvent conditions for each step (loading, washing, and elution). In addition, the introduction of trimethylaminoethyl (TMAE) derivatives to the hydroxyl group of the target analyte increased the ionization efficiency and sensitivity. The derivatized samples were analyzed using the LC-MS/MS method with electrospray ionization in the positive and multiple-reaction monitoring modes. The target analytes were separated and detected within 13.5 min using a CAPCELL PAK C18 MGII S3 column. Gradient elution was performed using distilled water and acetonitrile containing 5 mM ammonium acetate. This method offers a reliable and sensitive tool for the analysis of macroalgae samples for their potential use in biodiesel production. To the best of our knowledge, this is the first report on the simultaneous determination of fatty acids in macroalgae using LC-MS/MS with TMAE derivatization.

Exploration of Zero-Valent Iron Stabilized Calcium-Silicate-Alginate Beads' Catalytic Activity and Stability for Perchlorate Degradation.

Perchlorate contamination in groundwater poses a serious threat to human health, owing to its interference with thyroid function. The high solubility and poor adsorption of perchlorate ions make perchlorate degradation a necessary technology in groundwater contaminant removal. Here, we demonstrate the perchlorate degradation by employing nano zero-valent iron (nZVI) embedded in biocompatible silica alginate hybrid beads fabricated using calcium chloride (1 wt%) as a crosslinker. The concentration of precursors (sodium alginate, sodium silicate) for bead formation was standardized by evaluating the thermal stability of beads prepared at different sodium silicate and alginate concentrations. Thermal degradation of silica alginate hybrid samples showed a stepwise weight loss during the thermal sweep, indicating different types of reactions that occur during the degradation process. The formation of the silica alginate hybrid structure was confirmed by FT-IR spectroscopy. Scanning electron microscopy (SEM) data revealed the surface morphology of silica alginate hybrid changes by varying sodium silicate and alginate concentrations. nZVI-loaded alginate-silicate polymer bead (nZVI-ASB) exhibited excellent perchlorate degradation efficiency by degrading 20 ppm of perchlorate within 4 h. Our study also showed the perchlorate degradation efficiency of nZVI-ASB is maximum at neutral pH conditions.

Perspectives for biocatalytic lignin utilization: cleaving 4-O-5 and Cα-Cß bonds in dimeric lignin model compounds catalyzed by a promiscuous activity of tyrosinase.

BACKGROUND: In the biorefinery utilizing lignocellulosic biomasses, lignin decomposition to value-added phenolic derivatives is a key issue, and recently biocatalytic delignification is emerging owing to its superior selectivity, low energy consumption, and unparalleled sustainability. However, besides heme-containing peroxidases and laccases, information about lignolytic biocatalysts is still limited till date. RESULTS: Herein, we report a promiscuous activity of tyrosinase which is closely associated with delignification requiring high redox potentials (>1.4 V vs. normal hydrogen electrode [NHE]). The promiscuous activity of tyrosinase not only oxidizes veratryl alcohol, a commonly used nonphenolic substrate for assaying ligninolytic activity, to veratraldehyde but also cleaves the 4-O-5 and Cα-Cß bonds in 4-phenoxyphenol and guaiacyl glycerol-ß-guaiacyl ether (GGE) that are dimeric lignin model compounds. Cyclic voltammograms additionally verified that the promiscuous activity oxidizes lignin-related high redox potential substrates. CONCLUSION: These results might be applicable for extending the versatility of tyrosinase toward biocatalytic delignification as well as suggesting a new perspective for sustainable lignin utilization. Furthermore, the results provide insight for exploring the previously unknown promiscuous activities of biocatalysts much more diverse than ever thought before, thereby innovatively expanding the applicable area of biocatalysis.

Conversion of levulinic acid to 2-butanone by acetoacetate decarboxylase from Clostridium acetobutylicum.

In this study, a novel system for synthesis of 2-butanone from levulinic acid (γ-keto-acid) via an enzymatic reaction was developed. Acetoacetate decarboxylase (AADC; E.C. 4.1.1.4) from Clostridium acetobutylicum was selected as a biocatalyst for decarboxylation of levulinic acid. The purified recombinant AADC from Escherichia coli successfully converted levulinic acid to 2-butanone with a conversion yield of 8.4-90.3 % depending on the amount of AADC under optimum conditions (30 °C and pH 5.0) despite that acetoacetate, a ß-keto-acid, is a natural substrate of AADC. In order to improve the catalytic efficiency, an AADC-mediator system was tested using methyl viologen, methylene blue, azure B, zinc ion, and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) as mediators. Among them, methyl viologen showed the best performance, increasing the conversion yield up to 6.7-fold in comparison to that without methyl viologen. The results in this study are significant in the development of a renewable method for the synthesis of 2-butanone from biomass-derived chemical, levulinic acid, through enzymatic decarboxylation.

Association between precocious puberty and some endocrine disruptors in human plasma.

Endocrine disruptors that mimic natural hormones and inhibit the action of hormones have recently attracted attention as one of the main cause of precocious puberty. In this study, the levels of 7 EDCs and 3 isoflavones that exhibit estrogen-like actions were measured in the plasma of precocious puberty patients and compared to control subjects to determine if there is an association between the onset of precocious puberty and the levels of EDCs in the plasma. EDCs examined in this study were bisphenol-A (BPA), di(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP), mono(2-ethylhexyl) phthalate (MEHP), monobutyl phthalate (MBP), n-nonyl phenol (n-NP), and t-octylphenol (t-OP), and whereas the isoflavones were equol, genistein, and diadzein. The level of MBP in the plasma of patients was 1.3 times higher than that of the controls. The levels of t-OP and n-NP in the plasma of patients were respectively 1.15 and 1.2 times higher than those of the control group. Finally, the levels of the diadzein, equol and genistein were 1.37, 1.3 and 2.67 times higher than those of the control group, and genistein showed a statistically meaningful result (P = 0.0008). The results suggest that these six substances (MBP, t-OP, n-NP, daidzein, equol, and genistein) have an effect on precocious puberty.