Background: The search for new natural or synthetic products with antioxidant activity is commonly based on methods that involve reduction of either 2,2-diphenyl-1-picrylhydrazyl (DPPH) or 2-2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS). However, the reported values of the effective concentrations are highly variable, even in controls. Herein, we optimize and validate both methods of determining antiradical activity. Methods: Optimization was carried out using both a fractionated factorial design and a basic sequential simplex method, by monitoring the reduction percentage. Quercetin or Trolox were used as positive control. Furthermore, for each method, linearity, precision, accuracy, robustness, plate uniformity, signal variability, and Z factor, were established. Results: The optimized conditions for the DPPH method were: DPPH 280 µM in ethanol and 15 min of reaction time in the dark. The linear range was between 7 and 140 µM with an R2 value of 0.9987. The optimized conditions for the ABTS method were: ABTS adjusted to 0.7 absorbance units, 70% concentration in ethanol, and a reaction time of 6 min in the dark. The linear range was found to be between 1 and 70% with an R2 = 0.9991. For both methods, the accuracy and precision were within limits and the Z factor value was higher than 0.89. The applicability of each method was assessed by analyzing eight plant extracts. Conclusion: The DPPH and ABTS reduction methods were optimized and validated on a microscale and could be expected to be implemented in any laboratory.
Solid-phase microextraction (SPME) is a sample preparation technique with many applications that is being continuously developed. In this technique, the type of fiber coating plays a crucial role for extraction efficiency. Currently available commercial coatings have certain drawbacks that have been overcome by the development of new coatings based on novel materials; these have improved the efficiency of extraction, selectivity and stability of commercial coatings. Pharmaceutical and personal care products (PPCPs) are one of the most important groups of emerging contaminants; however, some studies suggest that these compounds can cause adverse health effects. No official monitoring protocols for these compounds are currently available, so the establishment of analytical methods that allow their determination in environmental samples is required. The complexity of environmental samples together with the low concentration levels of these compounds makes necessary the use of sample preparation techniques capable of removing interferences, as well as preconcentrated analytes, and SPME is a very promising alternative to achieve this. This review describes the recent developments in SPME with classical and novel coatings and its applications for PPCP determination in environmental samples.
The usefulness of traditional plants in Mexico to treat human ailments has been known since ancient times. This work evaluated the antimicrobial, anticoagulant, antioxidant, cytotoxic, and anti-inflammatory potential of ethanolic extracts of Aloe vera, Equisetum arvense, Mimosa tenuiflora, Lippia graveolens, and Syzygium aromaticum. The antimicrobial activity of the extracts was evaluated against Streptococcus mutans and Streptococcus sorbinus; a significant inhibitory effect of the L. graveolens extract on both bacteria was observed at concentration levels of 250 µg/mL and greater. The anticoagulant activity was evaluated in terms of prothrombin time (PT) and activated partial thromboplastin time (APTT), A. vera and M. tenuiflora extracts showed no significant difference (p ˂ 0.05) in PT compared with the control, and for APTT the extracts of A. vera, L. graveolens, and S. aromaticum decreased the APTT significantly (p ˂ 0.05) compared with the control. The antioxidant potential by DPPH assay indicated that the E. arvense extract behaved statistically the same as the control. The cytotoxic activity was evaluated in HGF-1 cells using the fluorometric microculture cytotoxicity assay technique, and none of the extracts was toxic at 125 and 250 µg/mL concentrations. Finally, the anti-inflammatory activity was evaluated using ELISA, where the A. vera extract showed the best anti-inflammatory capacity. Further research on the search for bioactive metabolites and elucidation of action mechanisms of the most promising extracts will be carried out.
7 February 2017 in Journal of Liquid Chromatography & Related Technologies
A new analytical temperature-assisted ionic liquid-based dispersive liquid–liquid microextraction (TA-IL-DLLME) method was developed for glyphosate and aminomethylphosphonic acid determination in water samples. Extracted analytes were derivatized using 9-fluoroenylmethylchloroformate and quantified by liquid chromatography with fluorescence detection. For the TA-IL-DLLME method, two strategies for phase solubilization were evaluated; in approach 1, the ionic liquid and aqueous matrix sample were mixed and then heated, while in approach 2, the aqueous sample was first heated and then the ionic liquid was injected. For both approaches, optimization included parameters that significantly affect extraction efficiency: ionic liquid type and volume, solubilization temperature and time, cooling and centrifugation time. Among the evaluated ionic liquids, 1-decyl-3-methylimidazolium tetrafluoroborate showed the best performance for TA-IL-DLLME and was selected for the two solubilization approaches; with approach 2, slightly better results were obtained. Thus, sample analyses were performed using a procedure based on approach 2. An important matrix effect, attributed to the presence of salts and metals in real water samples was observed. Sample acidification before derivatization allowed this problem to diminish, with recoveries ranging from 75 and 99%, and enrichment factors between 57 and 76 for target analytes. GRAPHICAL ABSTRACT
Eudragit® polymers have proven their potential as a means to control the release of aqueous insoluble drugs in various delivery systems as polymer nanoparticles (PNPs). The size (S) and polydispersity index (PDI) of PNPs are crucial factors for their interaction with biological systems from a pharmaceutical standpoint. This study aimed to determine the impact of the volumes of the organic phase (OP) and aqueous phase (AP), as well as the polymer amount (PA), on the size and PDI of PNPs prepared using the nanoprecipitation method for encapsulating quercetin (Qr). The study also evaluated the toxic effects of PNPs on human erythrocytes. The PNPs were prepared using preformed polymers derived from methacrylic acid and polyvinyl alcohol (PVA) as a surfactant. The nanoprecipitation technique enabled the production of particles smaller than 200 nm with a PDI lower than 0.2, and the study established the significant impact (p < 0.05) of the three variables related to the polymers and solvents. The selected PNPs contained 5 mg of Qr and 50 mg of Eudragit polymers (1:10 w/w Eudragit® EPO, E100, L100, and Eudragit L100-55) and diverse concentrations of PVA. The study found that including PVA in the AP increased the Qr encapsulation by up to 98%. The hemolytic potential of Eudragit® PNPs and Qr was assessed in human erythrocytes, with no significant cytotoxic activity observed (p < 0.001) compared with the control. In conclusion, via the nanoprecipitation technique, preparing PNPs with defined and homogeneous S to entrap the flavonol Qr efficiently is possible.
1 November 2018 in International Journal of Biological Macromolecules