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A sensitive and simple liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method for determination of dasatinib in rat plasma using one-step protein precipitation was developed. After addition of carbamazepine as internal standard (IS), protein precipitation by acetonitrile was used as sample preparation. Chromatographic separation was achieved on an SB-C18 (2.1 mm × 150 mm, 5 μm) column with methanol-0.1% formic acid as mobile phase with gradient elution. Electrospray ionization (ESI) source was applied and operated in positive ion mode; selective ion monitoring (SIM) mode was used to quantification using target fragment ions m/z 488.2 for dasatinib and m/z 338.7 for the IS. Calibration plots were linear over the range of 10–1000 ng mL−1 for dasatinib in rat plasma. Lower limit of quantification (LLOQ) for dasatinib was 10 ng mL−1. Mean recovery of dasatinib from plasma was in the range 82.2%–93.6%. Relative standard deviation (RSD) of intra-day and inter-day precision were both less than 8%. This developed method is successfully used in pharmacokinetic study of dasatinib in rats.

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Summary

Rapid high-performance liquid chromatographic methods with evaporative light scattering detection (HPLC-ELSD) and electrospray ionization multistage mass spectrometry (HPLC-ESI-MSn) have been established and validated for simultaneous qualitative and quantitative analysis of eight steroidal saponins in ten batches of Gongxuening capsule (GXN), a widely commercially available traditional Chinese preparation. The optimum chromatographic conditions entailed use of a Kromasil C18 column with acetonitrile-water (30:70 to 62:38, υ/υ) as mobile phase at a flow rate of 1.0 mL min−1. The drift tube temperature of the ELSD was 102°C and the nebulizing gas flow rate was 2.8 L min−1. Separation was successfully achieved within 25 min. LC-ESI-MSn was used for unequivocal identification of the constituents of the samples by comparison with reference compounds. The assay was fully validated for precision, repeatability, accuracy, and stability, then successfully applied to quantification of the eight compounds in samples. The method could be effective for evaluation of the clinical safety and efficacy of GXN.

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Acta Alimentaria
Authors: R.D. Wang, Y.J. Deng, L.J. Sun, Y.L. Wang, Z.J. Fang, D.F. Sun, Q. Deng, and R. Gooneratne

Growth and haemolytic activity of several pathogenic Vibrio species were compared in egg-fried-rice with different egg ratios. Egg-fried-rice preparations with rice-to-egg ratios of 4:1, 1:1, and 1:4 were inoculated with either Vibrio parahaemolyticus, V. cholerae, V. vulnificus, or V. alginolyticus and incubated for 24 h. Cell number, thermostable direct haemolysin (TDH) activity, and total haemolytic activity were determined. The cell number and total haemolytic activity increased in all Vibrio strains after 24 h, and these were most marked in egg-fried-rice with the highest egg content (1:4 (rice:egg) ratio; P<0.05). V. alginolyticus exhibited the maximal growth and V. parahaemolyticus the highest haemolytic activity, but only V. parahaemolyticus ATCC 33847, V. alginolyticus CAMT 21162, and V. alginolyticus HY 91101 showed TDH activity. Results suggest that lowering egg content in egg-fried-rice could reduce growth and virulence of Vibrio pathogens.

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Shewanella putrefaciens supernatant was found to increase the virulence factors of Vibrio parahaemolyticus by efficiently degrading its acylhomoserine lactone (AHL). To further reveal the regulation mechanism and its key degrading enzyme, a potential AHL-degrading enzyme acylase (Aac) from S. putrefaciens was cloned, and the influences of temperature, pH, protein modifiers, and metals on Aac were tested. Aac was significantly influenced by temperature and pH, and exhibited the highest AHL-degrading activity at temperatures of 37 °C and pH of 8. Mg2+ and Fe2+ can further increase the AHL-degrading activity. 10 mM EDTA inhibited its activity possibly by chelating the co-factors (metals) required for Aac activity. Tryptophan and arginine were identified as key components for Aac activity that are critical to its AHL-degrading activity. This study provides useful information on Aac and for V. parahaemolyticus control.

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In this study, a new substitution line, 12-5-1, with 42 chromosomes that was derived from BC3F2 descendants of the hybridization between Triticum aestivum cv. CN19 and Aegilops biuncialis was created and reported. The 12-5-1 was immune to both powdery mildew and stripe rust and has stable fertility. Multi-color fluorescence in situ hybridization indicated that 12-5-1 was a substitution line 1Mb(1B). The seed storage protein electrophoresis showed that 12-5-1 presented high molecular weight glutenin subunits (2 + 12) of CN19 and a new subunit designated as M which apparently originated from parent Ae. biuncialis, and absent 7 + 8 subunits. Additionally, the flour quality parameters showed that the protein content, Zeleny sedimentation value, wet gluten content, and grain hardness and mixing time of 12-5-1 were signifiantly higher than those of its parent CN19. Moreover, 5 pairs of the chromosome 1Mb-specifi polymerase chain reaction-based landmark unique gene markers, TNAC1021, TNAC1026, TNAC1041, TNAC1-02 and TNAC1-04, were also obtained. The new substitution line 1Mb(1B) 12-5-1 could be a valuable source for wheat improvement, especially for wheat end product quality and resistance to disease.

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