Sérgio Scherrer Thomasi

Human serum albumin-specific recognition of the natural herbal extract of Stryphnodendron polyphyllum through STD NMR, hyphenations and docking simulation studies

Over the last two decades, new and more advanced strategies that help in the rapid screening and identification of new ligands for a specific macromolecule have become an important domain. From this viewpoint, the effectiveness of STD NMR, Tr-NOESY, and STD-TOCSY has been utilized to evaluate the binding potential of the natural extract of Stryphnodendron polyphyllum, used as a herbal medicine in Brazil, towards human serum albumin. Moreover, 1D-DOSY experiments have also been carried out for the discriminations of different molecular weight compounds present in this extract. Following the STD, Tr-NOESY, and TOCSY analysis, a hyphenated system comprising LC-SPE-NMR was utilized to see the complete structural assignments through 2D spectra. The combined results from NMR spectroscopy and separation methods provided myricetin-3-O-rhamnopyranoside (1), quercetin-3-O-glucopyranoside (2), quercetin-3-O-xylopyranoside (3), and quercetin-3-O-rhamnopyranoside (4) as the active site blockers. Moreover, epitope results and additional Tr-NOESY cross peaks suggested the presence of the flattened conformations of these ligands within the ligand–HSA complex through the edge protons. Similarly, STD competition studies with the ligand–HSA complex were demonstrated by varying the concentration of spy molecule that selectively binds with Sudlows site II. Finally, docking simulations targeting both Sudlow sites (I and II) were performed, which interestingly mimic the STD competition results and showed that these compounds (1–4) are more prone towards binding site-1 inhibition. Therefore, we suggest that the sequence of techniques presented in this study can be considered as a simple and fast analytical tool for screening natural extracts to get better leads against any specific target.

Use of diffusion-ordered NMR spectroscopy and HPLC–UV–SPE–NMR to identify undeclared synthetic drugs in medicines illegally sold as phytotherapies

The informal (and/or illegal) e‐commerce of pharmaceutical formulations causes problems that governmental health agencies find hard to control, one of which concerns formulas sold as natural products. The purpose of this work was to explore the advantages and limitations of DOSY and HPLC–UV–SPE–NMR. These techniques were used to identify the components of a formula illegally marketed in Brazil as an herbal medicine possessing anti‐inflammatory and analgesic properties. DOSY was able to detect the major components present at higher concentrations. Complete characterization was achieved using HPLC–UV–SPE–NMR, and 1D and 2D NMR analyses enabled the identification of known synthetic drugs. These were ranitidine and a mixture of orphenadrine citrate, piroxicam, and dexamethasone, which are co‐formulated in a remedy called Rheumazim that is used to relieve severe pain, but it is prohibited in Brazil because of a lack of sufficient pharmacokinetic and pharmacodynamic information. Copyright

A validated (1)H NMR method for quantitative analysis of α-bisabolol in essential oils of Eremanthus erythropappus.

α-Bisabolol is a natural terpene produced by Eremanthus erythropappus and is widely used in cosmetics and pharmaceuticals due to its anti-inflammatory, antibacterial and antimycotic properties. Due to these applications, a control of composition and authenticity of commercial oils rich in this terpene is required, resulting in a demand for new methodologies for quality control. In this work a rapid and efficient method for quantification of α-bisabolol in the essential oil of E. erythropappus (candeia) using 1H NMR was developed, validated and compared to gas chromatography (GC) method. The quantification of α-bisabolol by 1H NMR was successfully achieved for most of the essential oil samples of E. erythropappus evaluated, except for those with a more complex composition. To circumvent this limitation a 2D NMR COSY contour map was used. This method proved to be a fast and efficient alternative, providing results with standard deviations SD<0.3%. All evaluated parameters (selectivity, linearity, accuracy/precision, repeatability, robustness and stability of analyte and internal standard in solution) gave satisfactory results. Using the 1H NMR signals at 5.36 and 5.13ppm, the limit of detection (LOD) and limit of quantification (LOQ) were 0.26 and 2.59mg, respectively. The results obtained by the 1H NMR method presented SD=0.59%, smaller than the value found for GC (SD=1.18%). Tukey tests have shown that the results obtained by 1H NMR and COSY methodology are similar to the obtained by the traditional GC-FID technique using external and internal standardization and normalization with 95% confidence.

Secondary metabolites from an infusion of Lippia gracilis Schauer using the LC-DAD-SPE/NMR hyphenation technique

The genus Lippia comprises approximately 200 species of herbs, shrubs and small trees, distributed throughout the South and Central America and tropical Africa. The species Lippia gracilis Schauer is an endemic aromatic plant of the Brazilian Northeast normally found in the states of Bahia, Sergipe and Piaui. The traditional communities of Caatinga, a semi-arid region of the Brazilian Northeast, use its leaves to treat throat and mouth infections, cutaneous diseases, ulcers, vagina disorders, acne, Pityriasis alba, dandruff, burns, wounds, sinusitis, bronchitis, nasal congestion, headache, jaundice and paralysis. Considering that the main investigations into the L. gracilis Schauer species are focused on its volatile constituents, in this report, we describe the isolation, by liquid chromatography with diode array detector-solid phase extraction/nuclear magnetic resonance (LC-DAD-SPE/NMR), of five flavonoids besides free and glycosidically bound carvacrol from an infusion of the leaves of a genotype of this species.

Bio- and Chemicalcontrol of Sclerotinia sclerotiorum using Stachybotrys levispora and its secondary metabolite griseofulvin

Sclerotinia sclerotiorum is responsible for the white mold of soybeans, and the difficulty to control the disease in Brazil is causing million-dollar damages. Stachybotrys levispora has shown activity against S. sclerotiorum. In our present investigation, we analyzed the chemical basis of this inhibition. Eight compounds were isolated, and using spectroscopic methods, we identified their structures as the known substances 7-dechlorogriseofulvin, 7-dechlorodehydrogriseofulvin, griseofulvin, dehydrogriseofulvin, 3,13-dihydroxy-5,9,11-trimethoxy-1-methylbenzophenone, griseophenone A, 13-hydroxy-3,5,9,11-tetramethoxy-1-methylbenzophenone, and 12-chloro-13-hydroxy-3,5,9,11-tetramethoxy-1-methylbenzophenone. Griseofulvin inhibited the mycelial growth of S. sclerotiorum at 2 μg mL-1. Thus, the antagonistic effect of S. levispora to S. sclerotiorum may well be due to the presence of griseofulvins. Our results stimulate new work on the biosynthesis of griseofulvins, to locate genes that encode key enzymes in these routes and use them to increase the production of these compounds and thus potentiate the fungicide effect of this fungus. S. levispora represents an agent for biocontrol, and griseofulvin represents a fungicide to S. sclerotiorum.

Synthetic acylsugars and their effects on the control of arthropod pests

One of the main problems facing agriculture is the loss of production as a result of the attack of agricultural pests. Alternative ways to work around this problem are being sought. There are substances called acylsugars that are naturally produced by the wild tomato species S. pennellii and affect arthropod pests. The objectives of this work were to synthesize two acylsugars and assess the biological effect of these on the arthropod pests Bemissia tabaci and Tetranycus urticae. The syntheses were performed via the reactions of glucose and sucrose (saccharose) with acetic anhydride using sodium acetate as the catalyst. The products of these reactions were sucrose octaacetate and glucose penta-acetate, the structures of which were confirmed by spectroscopic techniques. In a resistance test against the mite, a linear correlation between the concentration of the synthesized substances, and the dislocation of the mite was obtained. A delay in the hatching of the arthropod eggs was observed, causing a mortality rate of approximately 95% in the 1st instar larvae of mites that was confirmed in adults. In the biological tests with Bemisia tabaci, there was a low rate of hatching and emergence, and the effect on the nymphs was the deformation of the emergent adults.

Preparative Separation and Structural Identification of Impurities of a New α2-Adrenoceptor Agonist Using Stacking Injection, LC-MSn and LC-SPE-NMR

n ) and liquid chromatography-solid phase extraction-nuclear magnetic resonance (LC-SPE-NMR). The purification step was conducted using a semi-preparative liquid chromatography and stacked injections as a new approach to drug purification. As a result, a total yield of 75% of the pure LPSF-PT-31 and 2.9 L day -1 in solvent reduction was obtained. The combination of semi-preparative stacking injection, LC-MS n , and LC-SPE-NMR, demonstrated to be efficient to purify active drugs and unambiguously identify its impurities. In addition, isolation and identification of drug impurities in the early stages of development can improve the synthetic pathway, preventing the formation of impurities or minimizing this formation to minimum levels. n , LC-SPE-NMR, impurity identification