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Analyses were performed according to the modified method of Prasain et al. [11]. A C18 column 150 �� 4.6?mm (5?��m) (LichroCART, Merck, Germany) was maintained at 40��C, with a flow rate of 1?mL/min and detector fixed at 270?nm. The eluents were H2O acidified to pH 4.0 by 10?mM ammonium formate and formic acid (A) and acetonitrile (B). The following linear solvent gradient was applied: 100% A in 5?min, from 100% A to 80% A in 5?min, and a plateau of 10?min, to 60% A in 40?min. After separation, the HPLC effluent was delivered into a single quadrupole mass spectrometer (Agilent Technologies, USA) via orthogonal atmospheric pressure ionization-electrospray (API-ES) interface mode at 100�C700 m/z and step size at 0.2. The optimum electrospray ionization (ESI) conditions were as follows: ionization http://www.selleckchem.com/products/Thiazovivin.html mode, positive (4,000?V), negative (3,500?V); nebulizer pressure, 60?psi; drying gas flow rate, Icotinib 13?L/min; drying gas temperature, 320��C. 2.11. ABTS Assay The ABTS+ decolorization assay was performed according to the method described by Luximon-Ramma et al. [12], based on the ability of an antioxidant to scavenge the preformed ABTS+ radicals relative to that of the standard trolox. Results were expressed in mg trolox equivalents/g of extract in dry weight (mg TE/g dry wt). 2.12. DPPH Assay The DPPH radical scavenging assay was done according to the method of Brand-Williams et al. [13], based on the reduction of the stable radical, DPPH, to the formation of a nonradical form in the presence of hydrogen donating antioxidant. DPPH values were calculated using a linear regression equation of trolox YES1 standards, and the relative scavenging capacity of the extracts was expressed as mg trolox equivalent/g of extract in dry weight (mg TE/g dry wt). 2.13. FRAP Assay The FRAP assay was performed according to Luximon-Ramma et al. [12]. The reaction mixture was read at 740?nm every 5?min in 1st h, every 10?min in 2nd h, and every 20?min in 3rd h. Results were expressed as mg trolox equivalents/g of extract in dry weight (mg TE/g dry wt). 2.14. Statistical Analysis All the assays were performed in triplicate, and the results were expressed as mean �� SD from the three sets of observations. Comparisons were performed by one way ANOVA followed by Fisher's least significant difference (LSD) test using the SPSS 16.0 software (SPSS Inc., Chicago, IL, USA). P