VALIDATED STABILITY INDICATING HPLC APPROACH FOR QUANTIFYING TRICHOLINE CITRATE AND CYPROHEPTADINE SIMULTANEOUSLY IN SYRUP FORMS

Objective: This investigation demonstrates a stability-indicating and reliable “high-performance liquid chromatography” method to simultaneously quantify tricholine citrate (TEC) and cyproheptadine (CRH) in the syrup form and bulk form. Methods: Successful separation was accomplished using C18 “Agilent column (250 mm × 4.6 mm, 5 μm)” with isocratic type of elution using mobile phase containing 0.1 M NaH2PO4 buffer and acetonitrile at 55% volume and 45% volume ratio, respectively with 1.0 ml/min flow rate. The wavelength sensor was attuned at 263 nm to quantify TEC and CRH. Results: TEC and CRH peaks were eluted with fine resolution at retention times 1.837 min and 2.936 min, respectively. In the 137.5-412.5 μg/ml and 1-3 μg/ml concentration ranges for TEC and CRH, the calibration graphs were linear, with regression coefficients of 0.9999 and 0.9998, respectively. The suggested "high-performance liquid chromatography" approach has been shown as sensitive, precise, robust, accurate, specific and stability indicating through the resolution of TEC and CRH from its degradation-based compounds. Conclusion: The established high-performance liquid chromatography technique was effectively extended to the evaluation of TEC and CRH in the combined syrup form and the test results appeared satisfactory

Genetic diversity assessment of wild and cultivated varieties of Jatropha curcas (L.) in India by RAPD analysis

The present study deals with evaluation of genetic diversity and pedigree analysis through RAPD analysis. A total number of 40 Jatropha curcas accessions collected from different geographical regions and 43 random decamer primers were screened to assess polymorphism. 10 primers were amplified and 94 polymorphic bands were found out of 125 scored. Accounting for 75.2 % polymorphism across the genotypes 12.5 bands per primer, out of 9.4 were polymorphic. Jaccard’s coefficient of similarity varied from 0.00 to 1.00 indicative of high levels of genetic variation among the genotypes studied. Cluster analysis of data using UPGMA algorithm placed the 40 accessions into 2 main clusters, with cluster II divided into six sub-clusters. The result provides valid guidelines for the collection, conservation and characterization of Jatropha curcas genetic resources

Synthesis, spectral studies and antibacterial activity of iron(III) complexes with hydrazone functionalized ligands: X-Ray structure determination of a novel five coordinate complex containing labile ligands

608-615Iron(III) complexes having general formula FeLCl2 [where, L = 2-acetylpyridine acetoylhydrazone (APAH), 2-acetylpyridine benzoylhydrazone (APBH), 2-acetylthiophene acetoylhydrazone (ATAH), and 2-acetythiophene benzoylhydrazone (ATBH)] have been synthesized and characterized based on molar conductivity, electronic and IR spectroscopy. The structure of iron(III) complex with APBH ligand is determined using single crystal X-ray crystallography. The complex crystallizes in monoclinic space group P21/n with a = 7.8490(6) Å, b = 15.1018(11) Å, c = 13.2263(10) Å, α = 90°, β=100.183(3)°, γ = 90°, V = 1543.1(2) Å3 and Z = 4 with central Fe(III) ion coordinated by one tridentate APBH ligand. The iron is involved in 5-coordinate bonding with one organic (hydrazone) unit and two labile chloride ligands. The ligand acts as NNO–tridentate donor system. Iron is coordinated to pyridine ring nitrogen, azomethine nitrogen and benzoyl oxygen atoms and the two chloride ligands bind with metal completing distorted square pyramidal structure. The ligands and iron complexes are screened for their anti-bacterial activities against Pseudomonas aureoginos and Bacillus cereus. Among ligands, acetoyl hydrazones show more activity than the corresponding benzoyl hydrazones. The hydrazones having a methyl/ and pyridine groups show higher antibacterial activity. The iron complexes show higher activity than the metal free ligands

EVALUATING THE BEST POLYETHYLENE GLYCOL AS SOLID DISPERSION CARRIER BY TAKING ETORICOXIB AS A MODEL DRUG

Objective: The main objective of the current research is focused in discovering the best polyethylene glycol (PEG) as solid dispersion carrier using etoricoxib (ECB) as a model drug. Methods: Varieties of PEG, namely PEG - 3350, PEG - 4000, PEG - 6000, PEG - 8000, and PEG - 20000, were evaluated as a carrier for making ECB solid dispersions. ECB:PEG was taken in the ratios of 1:1, 1:2, 1:4, and 1:6. The solid dispersions were prepared by microwave fusion method and compressed using 8 station tablet compression machine. The fabricated solid dispersion tablets were tested for physicochemical characteristics and drug release rates. The release of ECB from the prepared solid dispersions was further analyzed kinetically using the first order and Hixson-Crowell’s plots. Results: All the solid dispersion batches were shown satisfactory physicochemical characteristics. ECB solid dispersion batches with PEG - 6000 showed good solubility in distilled water (up to 2.29±0.01 μg/ml) and in 0.1 N HCl (up to 2.18±0.01 μg/ml) when compared with ECB alone (0.21±0.01 μg/ml and 0.32±0.01 μg/ml). The prepared solid dispersions with PEG 6000 are shown good ECB release. Conclusion: Among PEG carriers, PEG - 6000 was found to be the best carrier for increasing the solubility and release rate of ECB form the solid dispersions compared to PEG - 3350, PEG - 4000, PEG - 8000, and PEG - 20000

VALIDATED GRADIENT STABILITY INDICATING RP-HPLC METHOD FOR THE SIMULTANEOUS QUANTIFICATION OF 11 RELATED SUBSTANCES IN THE COMBINED DOSAGE FORMS OF LAMIVUDINE AND TENOFOVIR DISOPEOXIL FUMARATE

Objective: Development of a stability-indicating reverse phase liquid chromatographic (RP-HPLC) method for the simultaneous quantification of 11 impurities in the combined dosage forms of lamivudine and tenofovir disoproxil fumarate drug substances.Methods: Efficient chromatographic separation of all analytes was achieved on a Waters X-terra RP18 column (150 x 4.6 mm, 3.5 mm) using mobile phase A (ammonium acetate buffer, pH adjusted to 5.0±0.05 with dilute orthophosphoric acid) and mobile phase B (mixture of methanol and ammonium acetate buffer in the ratio of 20:80) with the flow rate of 1.0 ml/min in gradient elution mode at 260 nm.Results: The method was validated in terms of the limit of detection, limit of quantification, linearity, accuracy, precision and robustness according to the international conference on harmonisation (ICH Q2R1). Regression analysis showed that the correlation coefficient (r2) is greater than 0.997 for individual active drug substances as well as their related substances. The method has proven very accurate (94.6 % to 108.2 % with % RSD not more than 4.9), highly precise (% RSD of the Intra-day and the inter-day study was not more than 8.9) and robust enough to deliver accurate results, when the chromatographic conditions were altered intentionally. Forced degradation studies were conducted in acidic, basic, thermal, photolytic, humid and peroxide stress conditions, where all the degradation peaks were monitored. Highest degradation of lamivudine was observed under oxidative stress condition and tenofovir was more susceptible to degradation under acidic and alkaline conditions.Conclusion: The present method is able to separate all the related compounds with each other and with the main drug substances with the resolution more than 2.0. The test solution was found to be stable in diluent up to 24 h. The mass balance of forced degradation of formulations, close to 99 %, made this method as a stability indicating method

METHOD DEVELOPMENT AND VALIDATION OF ERYTHROMYCIN AND OLAPARIB IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY–TANDEM MASS SPECTROMETRY

The liquid chromatography–tandem mass spectrometry (LC-MS)/MS methodology was used to develop and validate a method for detecting erythromycin and olaparib in human plasma. Antibiotics such as erythromycin and olaparib fall into this category. Liquid chromatography is used to separate stationary and mobile phases based on differences in their affinities as well as to remove unwanted contaminants. It improves repeatability, sensitivity, resilience, and low-level protein detection. A C18 (C18, 5 m, 100×4.6 mm) column is utilized for high resolution and peak area. The calibration curve is created using linear regression. Internally, telmisartan is utilized as a benchmark. The flow rate of the mobile phase is 0.5 mL/min. Erythromycin and olaparib have mass-to-charge ratios of 735.43–115.97 and 435.08–102.04, respectively. Erythromycin in combination with olaparib resulted in a 98% recovery rate. The precision and accuracy of the results determined by interday QC samples are within acceptable limits. There was no evidence of instability

METHOD DEVELOPMENT AND VALIDATION STUDY FOR QUANTITATIVE DETERMINATION OF GENOTOXIC IMPURITY AND ITS PRECURSOR IN FLUCONAZOLE SAMPLE BY LIQUID CHROMATOGRAPHY–TANDEM MASS SPECTROMETRY

Objective: The objective of this work is method development and validation study for quantitative determination of 1-[2-(2,4-difluorophenyl)-2,3-epoxypropyl]-1H-1,2,4-triazole, a genotoxic impurity and its precursor in a fluconazole drug sample by liquid chromatography–tandem mass spectrometry.Methods: LC-MS/MS analysis of these impurities was performed on Hypersil BDS C18 (100 mm x 4.0 mm, 3 µm) column. 5 mmol ammonium acetate and acetonitrile in the ratio of 65:35 (v/v) was used as the mobile phase with a flow rate of 0.4 ml/min. The developed method was accomplished with a short run time of 10 min. Triple quadrupole mass detector coupled with positive electrospray ionization was used for the quantification of genotoxic impurities in multiple reaction monitoring (MRM).Results: The method was validated as per International Conference on Harmonization (ICH) guidelines. The method was linear in the range of 0.30 µg/g to 11.37 µg/g for impurity A and 0.30 µg/g to 11.34 µg/g for impurity B with a correlation coefficient of 0.999. The accuracy of the method was in the range of 98.25 % to 100.53 % for both impurities.Conclusion: A specific, selective, highly sensitive and more accurate analytical method using LC-MS/MS coupled with positive electrospray ionization has been developed for the quantification of genotoxic impurity (1-[2-(2,4-difluorophenyl)-2,3-epoxypropyl]-1H-1,2,4-triazole) and its precursor (1-(2,4-difluorophenyl)-2-[1,2,4]triazol-1-yl-ethanone) at 0.3 µg/g with respect to the 5.0 mg/ml of fluconazole

PREVALENCE AND SEVERITY OF POSSIBLE DRUG-DRUG INTERACTIONS AMONG THE GERIATRIC PATIENTS AT AN INDIAN TERTIARY CARE TEACHING HOSPITAL

Objective: The objective of this study was to study the prevalence and severity of possible drug-drug interactions (DDIs) among the geriatric patients.Methods: The present study was a retrospective cross-sectional study. Case records of geriatric inpatients from the medical records department were included in the study and the case records of all the remaining age group inpatients were excluded from the study. All the collected cases were subjected to check for the DDIs using the software Micromedex 2.0 and were categorized into minor, moderate, and major based on the severity.Results: In this study, a total of 85 cases were screened for possible DDIs, and among them, 54 cases were found to be with 179 possible DDIs. The prevalence was observed to be 63.5%. Most of the possible DDIs were found to be with moderate severity (65.4%) followed by major (25.7%). Majority of the possible DDIs were observed in the Department of General Medicine (83.2%) followed by chest and tuberculosis (7.8%).Conclusion: Majorly, the severity of interactions was found to be moderate in this study. To reduce the DDIs, rationale prescriptions have to be prescribed by considering the risk-benefit ratio. Geriatrics should be prescribed very cautiously because the age-related pharmacokinetics plays a significant role. By taking all the above aspects into consideration, clinical pharmacist should play a crucial role in the prevention and management of DDIs, especially in geriatrics.Â

A STUDY ON THE PREVALENCE AND SEVERITY OF POSSIBLE DRUG-DRUG INTERACTIONS IN PEDIATRICS DEPARTMENT AT AN INDIAN TERTIARY CARE TEACHING HOSPITAL

Objective: To study the prevalence and severity of possible drug-drug interactions in the department of pediatrics.Methods: Case records of the in-patients of the pediatrics department from the medical records department were included and the records of the ambulatory patients were excluded from the study. All the collected cases were subjected to check for the drug-drug interactions by using the software micromedex 2.0 and the interactions were categorized based on the severity into minor, moderate and major.Results: A total of 142 cases were screened for possible drug-drug interactions (DDIs) and among them 76 cases were observed to be with possible DDIs.  The prevalence was found to be 53.5% in this study. Majority of the cases with possible DDIs were observed to be in females. Results of the age wise categorization revealed that majority of the possible DDIS were observed in children (2-12 y) followed by the infants (1 mo–2 y). The drug combinations amikacin+ampicillin, paracetamol+phenytoin and ofloxacin+ondansetron were found to be the frequently observed possible DDIs of minor, moderate and major severities respectively.Conclusion: Majority of the possible DDIs were of moderate severity followed by major. Clinical pharmacists should take the responsibility in assisting the pediatricians for screening the possible DDIs in the prescriptions there by preventing them and providing a better pharmaceutical care for the pediatric population. Â

Cloning and functional validation of early inducible Magnaporthe oryzae responsive CYP76M7 promoter from rice

Cloning and functional characterization of plant pathogen inducible promoters is of great significance for their use in the effective management of plant diseases. The rice gene CYP76M7 was up regulated at 24, 48, and 72 hours post inoculation (hpi) with two isolates of Magnaporthe oryzae Mo-ei-11 and Mo-ni-25. In this study, the promoter of CYP76M7 gene was cloned from rice cultivar HR-12, characterized and functionally validated. The Transcription Start Site of CYP76M7 was mapped at 45 bases upstream of the initiation codon. To functionally validate the promoter, 5′ deletion analysis of the promoter sequences was performed and the deletion fragments fused with the β-glucuronidase (GUS) reporter gene were used for generating stable transgenic Arabidopsis plants as well as for transient expression in rice. The spatial and temporal expression pattern of GUS in transgenic Arabidopsis plants and also in transiently expressed rice leaves revealed that the promoter of CYP76M7 gene was induced by M. oryzae. The induction of CYP76M7 promoter was observed at 24 hpi with M. oryzae. We report that, sequences spanning -222 bp to -520 bp, with the cluster of three W-boxes, two ASF1 motifs and a single GT-1 element may contribute to the M. oryzae inducible nature of CYP76M7 promoter. The promoter characterized in this study would be an ideal candidate for the overexpression of defense genes in rice for developing durable blast resistance rice lines