ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF NADIFLOXACIN BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Objective: In the present work, a rapid, precise and sensitive HPLC Method with UV detection (237 nm) for analysis of Nadifloxacin in Bulk was developed. Methods: Chromatography was performed with a mobile phase containing a mixture of 0.05 %v/v trifluoro acetic acid and acetonitrile (65:35 v/v) with flow rate 1.2 ml min-l. The proposed method was validated as per the standard guidelines. Result: The retention time was found to be 12.3 min. In the range of 0.03-5 ppm, the linearity of Nadifloxacin shows a correlation co-efficient of 0.9997. Percentage recovery of the drug was found to be good (98-102%). Validation of the developed method was successful for precision, robustness, specificity and selectivity and ruggedness. Conclusion: The developed HPLC method was found to be simple, sensitive, precise, accurate and reproducible and can be successfully used for the quantitative estimation of Nadifloxacin in bulk

STABILITY INDICATING METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS QUANTIFICATION OF SORAFENIB AND REGORAFENIB DRUG SUBTANCES BY USING RP-UPLC

Objective: The aim of the research work is to develop and validate a novel, sensitive, specific, rapid, accurate, precise and stability indicating gradient reverse phase ultra-performance liquid chromatography (RP-UPLC) method for the quantitative determination of sorafenib and regorafenib drug substances. Methods: Liquid chromatographic method used for the analysis of the anti-cancer drug substances like sorafenib and regorafenib and method was developed and validated by using efficient chromatographic separation method and was achieved with the use of acquity UPLC system was used consisting of quaternary pump, photodiode array detector an auto injector and on line degasser. Results: The separation was achieved using acquity UPLC BEH C18, 1.7 µm.2.1×50 mm analytical column at 30 °C employing a gradient elution. Empower software was used for data acquisition. During method validation all the parameters were evaluated as per ICH guidelines, which remained well within acceptable limits. Degradation of the drug substances was found to be stable to acidic, aqueous, basic hydrolysis, thermal hydrolysis and photolytic stress condition and the tests solution of the drug substance was found to be stable up to 24 h. Conclusion: The results of linearity, precision accuracy and specificity were proved to be within the limits. This method can be employed in routine analysis for simultaneous estimation of sorafenib andregorafenib drug substances in quality formulations and dissolution studies

ANALYTICAL APPLICATIONS OF SAFRANIN O

A simple and sensitive spectrophotometric method has been developed for the estimation of Mycophenolic acid. The method is based on the formation of   Ion-Association complex with MYCO formed an ion -association complex with basic dye, SafraninO. The cationic form of the dye SAFO involves in the formation of neutral coloured ion-association complexe with negative charge (acid groups in the drug) which is extractable into chloroform and behaves as a single unit being held together by electrostatic attraction. The absorption maxima were found to be at lMax 520 nm. The method obeys Beer's law within the limits 10-40µg/ml and gives reproducible results. Molar absorptivity value is obtained as8.424x104 L mol-1 cm-1 and recovery was found to be 99.17 ± 0.92 to 99.62 ± 0.27. Interferences of the other ingredients and excipients were not observed. The proposed method can be used for the determination of MYCO both in pure and pharmaceutical formulations. KeyWords: Mycophenolicacid (MYCO), SafraninO (SAFO), Ion- association Comple

A FAST AND SENSITIVE SPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF HYDRAZINE IN ATAZANAVIR SULFATE DRUG SUBSTANCE

Objective: To develop a fast and sensitive UV spectrophotometric method for the quantitative estimation of Hydrazine in Atazanavir Sulfate drug substances and validate as per ICH guidelines. Methods: The method was based upon the observation, that a characteristic colour results upon addition of a solution of p-Dimethylaminobenzaldehyde in ethyl alcohol and hydrochloric acid to hydrazine and estimated at absorbance maximum  458 nm in Atazanavir drug substance. Results: The developed method resulted in Hydrazine exhibiting linearity in the range 0.2 to 2.7 µg/g. The Intraday and interday precision is exemplified by relative standard deviation of 0.959 % and 0.947%. Percentage Mean recovery was found to be in the range of 97â€101%, during accuracy studies. The limit of detection (LOD) and limit of quantitiation (LOQ) were found to be 0.2 µg/g and 0.6 µg/g respectively. Conclusion: The present work was aimed to develop a visible spectrophotometric method, which is simple, sensitive, accurate and cost effective to evaluate the quality of the bulk and pharmaceutical formulations

DEVELOPMENT AND VALIDATION OF SEVEN PHENYL HYDRAZINE CHLORO ESTER ISOMERS (PGIs) BY RP-HPLC-UV METHOD IN ANTICOAGULANT DRUG SUBSTANCE; APIXABAN

Objective: The objective of this work was to develop and validate a simple and sensitive reverse-phase high-pressure liquid chromatography method for the determination of seven potential genotoxic impurities in Apixaban drug substance. Methods: The optimized separation was achieved by using ACE 3 C18 PFP (150 mm×4.6 mm, 3 µm) HPLC column. The mobile phase-A was a degassed mixture of 0.01M Ammonium acetate buffer(PH adjusted 4.9±0.05 with diluted glacial acetic acid) and mobile phase-B was a degassed mixture of Acetonitrile, Isopropyl alcohol and Buffer PH 4.9 in the ratio of 60:20:20 v/v/v. The gradient program was operated at a flow rate of 1.0 ml/min and UV detection was at 330 nm. Results: The method was superior at linearity for seven impurities and correlation coefficient values were larger than 0.999, moreover, in the separation point of view, this method further achieved no matrix interference through chromatography by better resolution of the other impurities from the Apixaban drug substance and its related impurities for the accurate analysis of seven potential genotoxic impurities. The established limits of detection (LOD), limits of quantification (LOQ) values for the seven mutagenic impurities were each of 5 ppm (0.015µg/ml) and15 ppm (0.045µg/ml) respectively. The developed method was validated as per ICH guidelines and applied as a generic method to determine these seven potential genotoxic impurities for the pharmaceutical process control and drug material release. Conclusion: Validation of this analytical method was carried out including stability, selectivity, linearity, accuracy, system precision, method precision and intermediate precision thus proving that the described RP-HPLC method could be employed for fast and simple analysis of sevenphenyl hydrazine chloro ester isomers in Apixaban drug substance

Brief note on infestation of Diplectanum sp. in Asian seabass

Recently open sea cage farming has emerged as an alternative and additional income source for fishermen and fish farmers in India. Asian seabass, Lates calcarifer is widely used in open sea cage culture due to its high market demand. In cage farming, high stocking densities and poor water quality enhance the parasite loads of the cultured fishes. In this study, 47 specimens of Asian seabass collected from cages located in Naganathwada, Sunkeri, Ankola were analysed with the aim of identifying the parasites prevalent among this species. All external and internal organs of each fish were examined separately under microscope for parasites. The collected monogenean parasites were washed in a 0.85% saline solution and fixed in 70% ethanol and identified. Most of the infected fishes had dark coloration of the body and postmortem findings revealed gills with excessive mucus secretion and sticking of the gill tips with greyish coloration

Argulus quadristriatus infestation in cage cultured Asian seabass

In Indian waters, genus Netuma is represented by two species namely N. bilineata and N. thalassina (Order: Siluriformes, Family: Ariidae). Rounded shout, thin lips, inconspicuous median longitudinal groove, and higher anal fin ray count (16-19) are characters of N. bilineata while N. thalassina has conical snout, clearly visible median longitudinal groove and lower anal fin ray count (13-15). Prior to the erection of N. bilineata (earlier considered as synonym of N. thalassina) as valid species, Indian workers had difference of opinion regarding the representation of species under this genus from Indian waters

Report on Amyloodinium spp. cysts infection in clownfish

A study was undertaken to record the occurrence of parasitic infections in ocellaris clownfish, Amphiprion ocellaris. Of a total eight A. ocellaris maintained in hatchery, three were found infected with different developmental stages of Amyloodinium spp. and were kept under observation

Compensatory growth and production economics of Silver pompano, Trachinotus blochii (Lacepede, 1801), fingerlings stunted by feed and space deprivation

The effect of stunting by feed and space deprivation on compensatory growth (CG) in Silver pompano, Trachinotus blochii, was investigated. A commercial pellet feed (45% protein and 10% fat) was fed two times a day, throughout the entire experiment. The 270-day experiment consisted of an initial 60-day stunting phase and a 60-day post-stunting phase carried out in 4 × 2 × 2 m3 galvanized iron (GI) rectangular cages, and a 150-day grow-out phase carried out in 3-m diameter circular GI cages. During the stunting phase, the normal fish (in triplicates) were stocked at lower stocking density (17 fish/m3) and fed at 10% of body weight (BW), while stunted fish (one replication) were stocked at about three times higher stocking density (56 fish/m3) and fed at a three times lower feeding rate (3% of BW). The stunted and normal fish were reared in triplicates during the post-stunting phase, at uniform stocking density (15 fish/m3) with feeding at a higher rate (10% of BW) for stunted fish and normal feeding rate (8% of BW) was adopted for normal fish. During the grow-out stage, each replication from the post-stunting phase was shifted to 3-m circular cages with the same feeding rates. The lag in growth in stunted fish (5.56 g against 9.43 ± 0.13 g of normal) during the stunting phase was compensated during the post-stunting phase (36.88 ± 2.23 g against 38.13 ± 1.48 g of normal) by higher feeding rate. There were no significant (p > 0.05) differences in final harvest, biometry, morphometry, dressing yield, carcass nutritional composition, and serum biochemical markers at the end of grow-out stage. Because of the significant difference (p < 0.05) in the total feed provided (5.2 kg for stunted fish against 22.8 kg for normal fish) and the lesser unit cost for the production of stunted fingerling (USD 0.087 for stunted fish against USD 0.106 for normal), the farming of stunted fish brought about a higher net operational revenue and benefit:cost ratio