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Human serum albumin (HSA) has pseudoesterase activity. So far on gel specific detection of such property of HSA is never reported. Moreover, protein binding dyes are non-specific for albumin. However, many of such dyes are used for HSA detection. So, dye-based albumin detection on the gel is expected to generate false-positive results for HSA. In this context, we have discovered that Fast Blue BB (FBBB, 0.12%) stains specifically HSA pseudoesterase activity with 2 Naphthyl acetate (2NA) as an ester substrate. Further, neostigmine has not inhibited the pseudoesterase activity associated with HSA. Neostigmine is a known inhibitor of many true esterases like acetylcholinesterase. So, neostigmine addition offers specificity to the method developed for staining of HSA. Additionally, 2NA stains HSA better than bovine serum albumin (BSA). Exploring all these novel findings, we have devised a simple method of HSA detection on the gel, accurately where other esterases are not detected. To the best of our knowledge, our method is the first to detect HSA pseudoesterase activity specifically on gel without getting interfered by any other esterase activity. The method detects HSA better than BSA. We feel that this method will go a long way for the specific detection of HSA on the gel. It is also relevant for understanding the purity of donor human milk matrix and pharmaceutical preparation of HSA. Our method can detect 7 μM of added HSA in human urine. Therefore, our method can be proceeded further for microalbuminuria detection in days to come.In developing countries, people mainly depend on rice as their primary source of calories. However, the thiamine content of rice is below minimal requirements. Biofortification, via genetic engineering, is a cost-effective strategy to increase thiamine content in rice. We report on the optimization of a matrix-specific method, including extensive optimization of the sample preparation to ensure maximal sensitivity and stability. The LC-MS/MS method was fully validated for the simultaneous quantification of thiamine, its precursors 4-methyl-5-(2-hydroxyethyl) thiazole (HET) and 4-amino-2-methyl-5-hydroxymethylpyrimidine (HMP) and its diphosphate derivative (TDP) in both polished and unpolished rice. Bias was below 9% for all analytes and total imprecision (CV%) was within pre-set acceptance criteria (≤15%) for both QCs and real samples. Thiamine monophosphate (TMP), for which no labeled analogue was available at the time of analysis, was determined without internal standard. Although both accuracy and precision criteria were met (bias and CV less then 12%), the determination of TMP was considered semi-quantitatively. Moreover, TMP was found to be only a minor thiamine form ( less then 1% of total thiamine in all lines analyzed, both wild-type and genetically engineered), with measurable levels only present in unpolished rice. Finally, the validity and applicability of the procedure were demonstrated via its successful application on rice lines, genetically engineered to enhance thiamine content. Consequently, this method allows to evaluate the success of biofortification strategies in rice.Conventional and sparse partial least squares-discriminant analysis (PLS-DA and sPLS-DA) have been successfully tested in order to authenticate avocado samples in terms of three different geographical origins and six kinds of cultivar. selleck chemicals llc For this, lipid chromatographic fingerprints of different avocado fruits have been acquired using gas chromatography coupled with flame ionization detector (GC-FID) and employed for building classification models. In addition, classification models concatenating strategy has been applied, which has proved to be successful to resolve multiclass problems in food authentication. Finally, fine performance metrics around of 0.95 were obtained for both multivariate classification methods.Among the physiological and pathological sulfur-containing species, cysteine (Cys) is the most typical one which is an important component of the REDOX system in vivo. Monitoring the level of Cys from other competing species seems quite important in pre-clinical diagnosis and therapeutic evaluation. Herein, we developed a selective fluorescent probe, BPCys, for Cys from the benzo[a]phenazin backbone which had the potential of anti-cancer potency. BPCys suggested advantages including high specificity (40 fold over other species), high sensitivity (detection limit 18 nM), wide pH adaptability (6.0-11.0) and in particular, the anti-cancer effect. Biological assays and in silico simulation hinted the potency of the detecting product on Topoisomerase I/II. In brief, this study raised a practical strategy for monitoring the Cys level in living cells, especially in cancer models with its anti-cancer potential, thus opened the mind of exploring more specific tool for specific applications.Because of the isomeric heterogeneity that is ubiquitous in analytical science, a formidable analytical challenge is to fully discriminate multiple isomers, especially those candidate isomers with various biological functions. Ion mobility mass spectrometry (IM-MS) has gained impressive advances for gaining molecular conformations, whereas coexisting structurally similar isomers often make unambiguous discrimination impossible due to the limited IM resolution of commercially available instruments. Herein, we demonstrate an energy-resolved collision-induced fingerprint (CIF) method to fully discriminate isomeric monosaccharide derivatives that differ in terms of composition, connectivity and configuration without complex instrument modifications. By simply increasing the collisional energy in the trap cell, the full width at half maximum (FWHM) of IM peaks can be markedly narrowed by at least 2-fold. Given the excellent reproducibility of CIF measurements, the full discrimination of isomers can benefit from their unique feature values and root-mean square deviation (RMSD) in CIF spectra. Moreover, rapid discrimination of each monosaccharide derivate isomer from binary mixtures is demonstrated. This strategy will expand the horizons of IM-MS platform in the rapid differentiation of a wider range of isomers more than monosaccharide derivatives in complex systems, which facilitates the identification and evaluation of innovative isomer candidates with unexplored functions.
My Website: https://www.selleckchem.com/products/paeoniflorin.html
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