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Coenzyme Q10 (CoQ10) is a cofactor in the respiratory chain and a potent endogenous antioxidant. Abnormally low levels of CoQ10 in the circulatory system are often involved with pathological states such as heart failure, mitochondrial and muscular diseases, and cancer. The development of simple, rapid, and highly sensitive methods capable of quantifying low CoQ10 levels in plasma is, therefore, increasingly required. In this work, we developed a miniaturized HPLC-UV system for the determination CoQ10 in human plasma using an XTerra microcolumn (50 mm × 2.1 mm i.d., 3.5 µm particle size), methanol:water (98:2, v/v) as mobile phase, 275 nm, flow rate of 0.3 mL/min, 30 °C column temperature, and 2 µL injection volume. The extraction procedure consists of a plasma precipitation with 1-propanol and evaporation under nitrogen to allow a 1.5-fold enrichment. The chromatographic analysis was accomplished in 7 min. As a result, it was possible to quantitate CoQ10 in small sample volumes down to 0.07 µM and detect as low as 0.02 µM in real plasma, with good accuracy and precision, even in pathological conditions. The proposed method was found to be suitable for routine CoQ10 determination in clinical laboratories. [ABSTRACT FROM PUBLISHER]

A simple, fast, and sensitive HPLC method with UV detection has been developed for the quantitation of omeprazole (OMZ) and major related substances in raw material and pharmaceutical formulation (paste) using a column of reduced length (50 mm) and diameter (2.1 mm) packed with hybrid particles. Chromatographic conditions were: 25°C, 1 µl injection volume, and UV detection at WV of 280 nm. The flow rate was 0.3 mL/min using methanol-phosphate buffer (pH 7.6) (40:60) as the mobile phase. Chromatographic purity was also determined with the same chromatographic conditions. The method was validated according to international guidelines (ICH guidelines) for specificity, linearity, LOD, LOQ, precision, accuracy, and robustness. The HPLC-UV method was found to be suitable for the quality control and stability studies of OMZ in a pharmaceutical formulation. [ABSTRACT FROM AUTHOR]

A simiple, rapid, and highly sensitive fast microbore HPLC method with UV detection has been developed for the quantitation of coenzyme Q10 (Co-Q10) and related substances in raw material, pharmaceutical, and cosmetic formulations using a column with hybrid particles. Chromatographic conditions were: 30ºC column temperature, 1 µL injection volume at 275 nm. The Co-Q10 analysis was performed within 2.5 minutes with a flow rate of 0.3 mL/min. with a mobile phase consisted of 100% methanol, and the related substances were separated with a gradient elution based on methanol-water. The developed method was validated for selectivity, linearity, precision, accuracy, and robutness. The method was found to be suitable for the quality control of Co-Q10 in pharmaceutical and cosmetic products, as well as the stability indicating studies. [ABSTRACT FROM AUTHOR]

Smallanthus sonchifolius (Poepp. & Endl.) H. Robinson, commonly known as yacon, is a medicinal plant belonging to the Asteraceae family used in traditional folk medicine. Its roots and leaves have been used by people suffering from diabetes or from various digestive or renal disorders. This study aimed at evaluating the in vitro potential genotoxic effects of the aqueous extract of yacon in order to determine its safety and at characterizing its phytochemical composition. The aqueous extract of S. sonchifolius was prepared in a similar way to that commonly used in popular medicine as tea bags. Thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC-MS/MS) were used to identify the main compounds. The MTT test was performed to determine the range of doses and the Cytochalasine B-blocked micronucleus (Cytome assay) was used to assess geneotoxicity. The chemical analysis of the aqueous extract revealed the presence of the sesquiterpene lactones (STLs) enhydrin and the dimer enhydrofolin, as the main compounds together with phenolic compounds. Increasing concentrations of the extract induced a cytotoxic effect on CHO–K1 and HepG2 cells. A statistically significant increase in the frequency of MNi, NBUDs and NPBs was observed in CHO–K1 cells, while in HepG2 cells a statistically significant frequency increase was observed with three of the four tested doses for MNi and only with the highest dose for NPBs and NBUs (genotoxic effect). Results demonstrated the inability of the metabolic system to counteract the genetic instability, allowing the safe consumption of the leaves as a 2% tea infusion in quantities of up to 250 mL/day. Image 1 [ABSTRACT FROM AUTHOR]

Extracts of Smilax campestris Griseb (Smilacaceae) have been employed in the treatment of several inflammatory diseases as a traditional herbal medicine. However, the cellular and molecular mechanisms involved in the observed effects remain elusive. Macrophages are known to play a central role in inflammatory responses. These cells are activated in response to a diversity of danger signals and produce several mediators of inflammation that eventually regulate the immune response. For all the above mentioned, scientific evidence is required to support the popular use of S. campestris. We aimed to investigate the anti-inflammatory effect of S. campestris aqueous extract (SME) in activated THP-1 human macrophages, on the production of some mediators of inflammation and oxidative stress in order to provide scientific support for its popular use. The characterization of SME was assessed by HPLC-MS/MS. The production of the pro-inflammatory cytokines and chemokines was evaluated by ELISA. The activity of metalloproteases was evaluated by zymography. The subcellular localization of the NF-κB transcription factor was analysed by Western blot. The superoxide anion and glutathione levels were assessed by flow cytometry. The cytotoxicity induced by SME in THP-1 macrophages was also investigated by the LDH release test. In the present study, we have identified catechin and glycosylated derivatives of quercetin (quercetin-3-O-glucoside, quercetin-3-O-galactoside, rutin and quercetin-3-rhamnoside) as major components of the aqueous SME. We found that SME significantly decreased the production of the pro-inflammatory cytokines tumour necrosis factor (TNF)- α, interleukin (IL)-1β, IL-6, IL-8 and monocyte chemoattractant protein (MCP)-1 and the activity of the metalloproteinase (MMP)-9, in lipopolysaccharide-activated macrophages derived from the monocytic cell line THP-1. Furthermore, SME diminished the expression of NF-κB p65 subunit in the nuclear fraction. In addition, SME decreased the production of superoxide anion in THP-1 macrophages, without altering the levels of reduced glutathione. These results suggest that SME exerts its anti-inflammatory effects in human activated macrophages by inhibiting the production of pro-inflammatory cytokines, matrix metalloproteinases and the NF-κB transcription factor pathway along with a reduction of oxidative stress mediators. Moreover, catechin and glycosylated derivatives of were identified by HPLC-MS/MS in SME. Our findings provide scientific support for the traditional use of the S. campestris extracts. Image 1 [ABSTRACT FROM AUTHOR]