Development and validation of a RP-HPLC method for determination of citicoline monosodium in human plasma

A sensitive and specific high performance reversed phase liquid chromatographic method was developed for quantification of citicoline monosodium (CTM) in human plasma. The active drug was isocratically eluted at a flow rate of 1 ml/min at ambient temperature in a nucleosil C18 analytical column with a mobile phase composed of tetrabutyl ammonium hydro gen sulfate buffer (0.005 M, pH5.0): methanol (95:05, v/v). Photodiode array (PDA) was performed at 270 nm and the retention time of the drug was found to be 6.64 min. The lowest limit of quantification (LLOQ) and of detection (LOD) were found to be 30 and 10 ng/ml, respectively. The method was validated and the response was found to be linear in the drug (CTM in spiked plasma) concentration range 150-900 ng/ml. The method was found to be accurate, with ranging from 96.38 to 98.65 % and precise, with intra-day, inter-day as well as analyst-toanalyst precision. The total recoveries of the method ranged between 95.69 and 97.89 %. Stability data revealed that the drug is stable in human plasma under various test conditions and the method can be successfully used for analysis of CTM in human plasma and in pharmacokinetic studies.Colegio de Farmacéuticos de la Provincia de Buenos Aire

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Not AvailableAmong the twenty-one bacterial endophytes isolated from Holy basil (Ocimum tenuiflorum), eight were potentially inhibiting the growth of five plant pathogens. Percent suppression of Rhizoctonia solani, Sclerotium rolfsii, Alternaria alternata, Microphomina phaseolina, and Bipolaris sorokiniana radial growth was significantly higher by isolates BTL-4 (70.64%), BTL-1 (80.63%), BTL-1 (57.50%), BTL-5 (81.28%), and GTS-15 (73.81%), respectively. Based on 16S rRNA gene sequencing, these isolates were putatively identified as Bacillus altitudinis (BTL-1 and GTS-16), Bacillus tequilensis (BTL-4), Bacillus safensis (BTL-5), Bacillus haynesii (GTR-8), Bacillus paralicheniformis (GTR-11), Bacillus pacificus (GTR-12), and Bacillus siamensis (GTS-15). Selected endophytes were tested against R. solani in vivo and found to reduce sheath blight disease incidence to a varying extent. Rice plants challenged with R. solani and inoculated with Bacillus altitudinis GTS-16 exhibited the least value of percent infected tillers, recorded maximum induction of defense-related enzymes (phenyl ammonia lyase, peroxidase, and polyphenol oxidase), and enhanced dry matter accumulation. Confocal Scanning Laser Microscope imaging using LIVE/ DEAD™BacLight™ bacterial viability staining has indicated trans-genera colonization of O. tenuiflorum endophytes in rice. Exploring the potential niches like the endosphere could open the vast arena of opportunity for developing newer strains of biocontrol agents.Not Availabl

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Not AvailableBackground: Ralstonia solanacearum has the problem of losing the virulence in laboratory conditions, during prolonged experimentation. Since pure colonies of R. solanacearum contain cell fractions differing in virulence, it was considered worthwhile to find a way of selecting the cells with lower attenuation. Therefore, a methodology for inducing virulent-type colonies occurrence in Ralstonia solanacearum was developed. Methods: Nutrient gradient was created by swabbing R. solanacearum culture in a slanted KMTTC medium, and Phyllanthus emblica extract was given by well diffusion. Live–dead cell imaging using BacLight, effects of ascorbic acid on cell viability, and production of virulence factors (exopolysaccharides, cellulase, and pectinase) supported this hypothesis. The tagging of R. solanacearum with green fluorescent protein and further confocal scanning laser microscopic visualization confirmed the colonization in vascular bundles of tomato. Results: P. emblica extract suppressed R. solanacearum initially in well diffusion, but further developed virulent-type colonies around the wells. Nutrient deprivation was found to have synergistic effects with P. emblica extract. The converted fluidal (virulent type) colonies could be able to colonize vascular bundles and cause wilting symptoms. Conclusion: This method will be useful in the laboratories working on biocontrol of R. solanacearum for maintaining virulent-type colonies. Moreover, it could form the basis for studies on the stability of phenotypic conversion and cell fractions in R. solanacearumNot Availabl