VALIDATED HPLC-UV METHOD FOR SIMULTANEOUS ESTIMATION OF LINAGLIPTIN AND EMPAGLIFLOZIN IN HUMAN PLASMA

Objective: The proposed method aims to develop a simple, rapid, sensitive and validated isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of linagliptin and empagliflozin in human plasma.Methods: Chromatography was performed on waters 2695 HPLC equipped with a quaternary pump. The separation was carried using discovery C18 (250×4.6×5) column, buffer: acetonitrile (68:32) as mobile phase with 1 ml/min flow rate. The analyte detection was monitored at 218 nm.Results: Retention time of linagliptin, empagliflozin and internal standard was found at 6.421, 4.696, and 4.074 min respectively. The peaks were found to be free of interference. The method is validated over a dynamic linear range of 0.01-10.0 µg/ml for both drugs with a correlation coefficient of 0.998. The precision and accuracy of samples of six replicate measurements at lower limit of quantification (LLOQ) level were within the limit. The analytes were found to be stable in human plasma at-28 °C for 37 d.Conclusion: The stability, sensitivity, specificity and reproducibility of this method make it suitable for the determination of linagliptin and empagliflozin in human plasma

ISOLATION OF ANTIBIOTIC PRODUCING BACTERIA FROM POND SOIL, GUDLAVALLERU

Soil being a major reservoir for microorganisms it is a source of interest for isolation of antibiotic producing organisms. The emergence of antibiotic resistance and need for better, broad spectrum antibiotics is always in high demand. In the present study, antibiotic producing bacteria were isolated from a local soil sample. Total ten soil samples were collected from local pond aseptically and subjected to serial dilution. Crowded plate technique was employed for the isolation of the colony. Total five isolated were isolated which exhibited zone of inhibition around the colony. The isolated colonies were subjected to morphological, microscopical and biochemical characterization. All five colonies were found to be gram positive, non-sporulating organisms and found they belong to the Actinobacteria class. The isolated colonies were subjected to screening for antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and Yeast by perpendicular streak method. The primary screening results conclude that except one colony all have good antimicrobial activity. One colony found to be highly potential activity which had inhibition towards gram positive, gram negative, sporulating and fungal activity. This study may contribute in providing information on the antibiotic producing microorganisms in soil. Further characterization, purification, and structural elucidation are recommended to know the novelty, quality and commercial value of these antibiotics

HPLC method development and validation for the estimation of axitinibe in rabbit plasma

<div><p>ABSTRACT A rapid, sensitive, and accurate high performance liquid chromatography for the determination of axitinibe (AN) in rabbit plasma is developed using crizotinibe as an internal standard (IS). Axitinibe is a tyrosine kinase inhibitor, used in the treatment of advanced kidney cancer, which works by slowing or stopping the growth of cancer cells. The chromatographic separation was performed on a Waters 2695, Kromosil (150 mm × 4.6 mm, 5 µm) column using a mobile phase containing buffer (pH 4.6) and acetonitrile in the ratio of 65:35 v/v with a flow rate of1 mL/min. The analyte and internal standard were extracted using liquid-liquid extraction with acetonitrile. The elution was detected by photo diode array detector at 320 nm.The total chromatographic runtime is 10.0 min with a retention time for axitinibe and IS of 5.685, and 3.606 min, respectively. The method was validated over a dynamic linear range of 0.002-0.2µg/mL for axitinibe with a correlation coefficient of r2 0.999.</p></div