Clayton Pedersen (bowlcicada68)

Isoflavonoid phytoestrogens, referred as "dietary estrogens" are widely distributed in the plant kingdom. Formononetin, biochanin A and their active metabolites daidzein and genistein are known to be the most potent among other isoflavonoid phytoestrogens. Thus there is a growing need to determine accurately their concentration in different biological fluids. In the present work, a sensitive analytical method was developed for the quantitative determination of these compounds in human breast milk, saliva and urine. The glycoside conjugates of these compounds were enzymatically hydrolysis prior to salting-out assisted liquid-liquid extraction. Quantitative analysis was done by ultra high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. The obtained results showed high correlation coefficients (r2 > 0.998) for the linear range established for formononetine, biochanin A, daidzein and genistein. The limits of detection (LODs) and low limits of quantitation (LLOQs) wuction status in human. In the current study, Pesticide residue extraction in beeswax was carried out using a mixture of acetonitrile-ethyl acetate (13, v/v). This mixture of solvents not only enables the melting of beeswax sample at a lower temperature than when using acetonitrile only but also introduces one phase solution. The sample extract was directly injected into both GC-MS/MS, of the commonly used split-less inlets, and into LC-MS/MS. Sample preparation and clean-up were also optimized. The developed method was validated according to SANTE/11813/2017 European Union guidelines. Three spiking levels of low concentrations 20, 50, 100 µg/kg were studied for the analysis of a total of 373 pesticides. Most of the studied pesticides have acceptable recovery between 80 and 110% with good reproducibility less then 10. There are 265 and 139 pesticides having a lower limit of quantifications equal 20 µg/kg using LC-MS/MS and GC-MS/MS, respectively. Finally, the developed method was successfully applied for the analysis of real beeswax samples. Three normal phase HPLC methods were produced to separate lipid classes on a PVA-Sil stationary phase including 9 polar lipids (method 1); 13 combined polar and neutral lipids (method 2); and a combined method that further separates the neutral lipids into 2-4 subclasses based on the presence of fatty acids containing a polar functional group (e.g. hydroxyl) for a total of 20 lipid classes and subclasses separated in a single run (method 3). Polar lipids separated include the phosphoglycerolipids PG, PE, PI, PS, PC and LPC; the galactoglycerolipids MGDG and DGDG; and a sulfoglycerolipid SQDG. Neutral lipids include TAG, DAG, and MAG classes and sub-classes containing 0-3, 0-2, and 0-1 hydroxy fatty acids, respectively. The hexane/isopropanol/methanol/aqueous system separates polar lipids without the use of chloroform such that it is suitable for radioactivity analysis by in-line flow scintillation counting. Each method was optimized using the natural lipid standards comprised of diverse molecular species that were detected by ELSD. All molecular species of each lipid class eluted together as single peak detected by ELSD. The methods were demonstrated to be suitable for resolving lipid extracts from animal, microbial, and plant sources as well as application to 14C based metabolic tracing of lipid metabolism in leaves and seeds. As part of the "omics" technologies in the life sciences, metabolomics is becoming increasingly important. In untargeted metabolomics, unambiguous metabolite identification and the inevitable coverage bias that comes with the selection of analytical conditions present major challenges. Reliable compound annotation is essential for translating metabolomics data into meaningful biological information. DBr-1 Here, we developed a fast and transferable method for generating in-house MS2 libraries to improve metabolite identification. Using