Yerba Mate (Ilex paraguariensis) Processing and Extraction: Retention of Bioactive Compounds.

Ilex paraguariensis is a native tree from South America known for the presence of bioactive compounds, and its processed leaves are consumed as hot and cold infusions. After harvest (step 1), the leaves are subjected to flame blanching to inactive the enzymes (step 2), followed by drying and milling (step 3). The impacts of I. paraguariensis processing on leaf composition were investigated by extracting the major compounds (chlorogenic and isochlorogenic acids (3-CQA, 4-CQA, 5-CQA, 3,4-DQA, 3,5-DQA and 4,5-DQA), p-coumaric acid, caffeine and rutin) using different ratios of ethanol and water as extraction solvent (EW 25:75, 50:50, and 75:25 (w/w)). The solvent ratio of EW 50:50 was more effective in extracting the chlorogenic acids isomers, with retention of chlorogenic acids of 3463, 9485, and 9516 µg mL- 1 for steps 1, 2, and 3, respectively. Rutin and p-coumaric acid exhibited similar behavior with the increment of processing steps; however, p-coumaric acid was only detected in steps 2 and 3 for the solvent ratios EW 50:50 and 25:50. The caffeine extraction from I. paraguariensis varied from 936 to 1170 µg mL- 1 for all processing steps, with emphasis on its concentration extracted in step 1. The evolution of processing steps led to a higher retention of phenolic compounds from I. paraguariensis, which was not observed when using different solvent ratios, and the solvent ratio EW 50:50 was more effective for the extraction of chlorogenic acids. The successful extraction of chlorogenic acids from I. paraguariensis in this study proved to be a promising alternative for the use of yerba mate beyond the cuia cup.

Development and validation of liquid chromatography-tandem mass spectrometry method to quantify dasatinib in plasma and its application to a pharmacokinetic study

Abstract Dasatinib, a potent oral multi-targeted kinase inhibitor against Src and Bcr-Abl, can decrease inflammatory response in sepsis. A simple and cost-effective method for determination of an effective dose dasatinib was established. This method was validated in human plasma, with the aim of reducing the number of animals used, thus, avoiding ethical problems. Dasatinib and internal standard lopinavir were extracted from 180 uL of plasma using liquid-liquid extraction with methyl tert-butil ether, followed by liquid chromatography coupled to triple quadrupole mass spectrometry in multiple reaction monitoring mode. For the pharmacokinetic study, 1 mg/kg of dasatinib was administered to mice with and without sepsis. The method was linear over the concentration range of 1-98 ng/mL for DAS in mice and human plasma, with r2>0.99 and presented intra- and interday precision within the range of 2.3 - 6.2 and 4.3 - 7.0%, respectively. Further intra- and interday accuracy was within the range of 88.2 - 105.8 and 90.6 - 101.7%, respectively. The mice with sepsis showed AUC0-t = 2076.06 h*ng/mL and Cmax =102.73 ng/mL and mice without sepsis presented AUC0-t = 2128.46 h*ng/mL. Cmax = 164.5 ng/mL. The described analytical method was successfully employed in pharmacokinetic study of DAS in mice.

Pretreatment and non-specific binding in ultrafiltration device: Impact on protease inhibitor quantification.

INTRODUCTION: Ultrafiltration (UF) is used to separate unbound drugs; however, non-specific binding (NSB) may be a limiting factor of this technique. Pretreatment of UF devices has been suggested to reduce NSB. Therefore, the pretreatment methodologies for UF devices were evaluated in order to test their effectiveness in reducing NSB of protease inhibitors (PIs). METHODOLOGY: Two PIs (lopinavir-LPV and ritonavir-RTV) were tested. UF devices were pretreated with ultrapure water, Tween-20 or Tween-80. To evaluate the NSB, after UF devices being pretreated, ultrafiltrate solutions containing the analytes at two concentrations (low and high) were used. Samples were quantified by LC-MS/MS. RESULTS: UF devices pretreated with Tween-5% had the lowest NSB for both analytes. NSB values varied between 7 and 11% at low concentration 16-34% at high LPV concentration, respectively. For RTV, NSB was approximately 6% for low concentration and 18% for high concentration. Failure to completely remove Tween in UF devices could results in an overestimation of NSB. CONCLUSION: Pretreatment of UF device with Tween and subsequent removal proved to be effective in reducing NSB of PI.

The Yin and Yang of Tyrosine Kinase Inhibition During Experimental Polymicrobial Sepsis.

Neutrophils are the first cells of our immune system to arrive at the site of inflammation. They release cytokines, e.g., chemokines, to attract further immune cells, but also actively start to phagocytose and kill pathogens. In the case of sepsis, this tightly regulated host defense mechanism can become uncontrolled and hyperactive resulting in severe organ damage. Currently, no effective therapy is available to fight sepsis; therefore, novel treatment targets that could prevent excessive inflammatory responses are warranted. Src Family tyrosine Kinases (SFK), a group of tyrosine kinases, have been shown to play a major role in regulating immune cell recruitment and host defense. Leukocytes with SFK depletion display severe spreading and migration defects along with reduced cytokine production. Thus, we investigated the effects of dasatinib, a tyrosine kinase inhibitor, with a strong inhibitory capacity on SFKs during sterile inflammation and polymicrobial sepsis in mice. We found that dasatinib-treated mice displayed diminished leukocyte adhesion and extravasation in tumor necrosis factor-α-stimulated cremaster muscle venules in vivo. In polymicrobial sepsis, sepsis severity, organ damage, and clinical outcome improved in a dose-dependent fashion pointing toward an optimal therapeutic window for dasatinib dosage during polymicrobial sepsis. Dasatinib treatment may, therefore, provide a balanced immune response by preventing an overshooting inflammatory reaction on the one side and bacterial overgrowth on the other side.

Atividade antioxidante e antimicrobiana de Calceolaria chelidonioides Humb. Bonpl. & Kunth. / Antioxidant and anti-microbial activity from Calceolaria chelidonioides Humb. Bonpl. & Kunth.

A espécie Calceolaria chelidonioides (Scrophulariaceae), até então inédita nas citações científicas, foi estudada sob o ponto de vista farmacológico buscando-se identificar possíveis atividades antimicrobiana e antioxidante em metodologia in vitro. As partes aéreas dessa espécie demonstraram atividade antioxidante em modelo usando o radical livre DPPH. As flores de C. chelidonioides mostraram grande potencial antibacteriano frente à bactéria Staphylococcus aureus resistente a meticilina MRSA, um dos principais responsáveis em casos de infecção hospitalar.

The species Calceolaria chelidonioides (Scrophulariaceae), not scientific described so for, was studied in pharmacological aspects aiming to identify some anti-microbial and antioxidant activity. The aerial parts showed antioxidant activity using in vitro DPPH model. The flowers from C. chelidonioides showed strong antibacterial potential against meticiline resistant Staphylococcus aureus (MRSA) strains the main responsible for hospital infection complications.