Development and Validation RP-HPLC Method for Simultaneous Estimation of Dapagliflozin and Saxagliptin in Bulk and Pharmaceutical Dosage Form

A simple, precise, sensitive, and rapid reverse phase high performance liquid chromatographic method was developed and validated for simultaneous estimation of Dapagliflozin (DAPA) and Saxagliptin (SAXA) in bulk as well as in tablet formulation according to the ICH guidelines. The chromatographic phase consisted by phosphate buffer: Acetonitrile, pH 4.0 adjusted by glacial acetic acid. The flow rate was adjusted to 0.8 ml/min and UV detection was carried out at 220 nm. Retention time was found to be 2.144 and 3.156 min; respectively. The detector was showed that linear responses over the concentration range 10–24 μg/ml for SAXA and 12–40 μg/ml for DAPA a good correlation coefficient of 0.999. This proposed method is highly sensitive, precise, and accurate which reduces cost of analysis; hence, recommended for routine quality analysis in laboratories

Development and Validation RP-HPLC Method for Simultaneous Estimation of Bilastine and Montelukast in Bulk and Pharmaceutical Dosage Form

A simple, precise, sensitive, and rapid reverse phase high-performance liquid chromatography method was developed and validated for simultaneous estimation of bilastine and montelukast in bulk as well as in tablet formulation according to ICH guidelines. The chromatographic phase consisted by methanol and acetonitrile (70:30) at pH 3 adjusted by 0.1% orthophosphoric acid. The flow rate was adjusted to 1 ml/min and ultraviolet detection was carried out at 260 nm. The retention time for of bilastine and montelukast were found to be 3 and 7 min, respectively. The detector was showed linear responses over the concentration range 25–150 μg/ml for bilastine and 5–30 μg/ml for montelukast a good correlation coefficient of 0.999. This proposed method is highly sensitive, precise, and accurate which reduces cost of analysis, hence recommended for routine quality analysis in laboratories

A Simple HPLC-UV Method for the Determination of Glutathione in PC-12 Cells

A highly sensitive and simple HPLC-UV method was developed and validated for the assay of glutathione (GSH) in PC-12 cells. Glutathione is a major intracellular antioxidant having multiple biological effects, best known for its cytoprotective effects against cell damage from reactive oxygen species and toxic reactive metabolites and regulating the cellular redox homeostasis. Due to its own sulfhydryl (SH) group, GSH readily reacts with Ellman’s reagent to form a stable dimer which allows for quantitative estimation of GSH in biological systems by UV detection. The separation was achieved using a C8 column with a mobile phase consisting of phosphate buffer adjusted to pH 2.5 (mobile phase A) and acetonitrile (mobile phase B), running in a segmented gradient manner at a flow rate of 0.8 mL/min, and UV detection was performed at 280 nm. The developed HPLC-UV method was validated with respect to precision, accuracy, robustness, and linearity within a range of 1–20 μg/mL. Limit of detection (LOD) and limit of quantification (LOQ) were 0.05 and 0.1 μg/mL, respectively. Furthermore, the method shows the applicability for monitoring the oxidative stress in PC-12 cells

Detection of Enterobacteriaceae producing CTX-M extended spectrum β-lactamases from a tertiary care hospital in south India

A total of 23 clinical isolates (15 Escherichia coli and 8 Klebsiella pneumoniae ), resistant to cefotaxime and ceftazidime recovered during 2002 and 2003, were investigated for production of CTX-M extended spectrum β-lactamase (ESBL) by phenotypic and molecular methods. The presence of ESBL was tested by NCCLS phenotypic confirmatory test using cephalosporin/clavulanate combination discs and E-test ESBL strips. Determination of MIC of cefotaxime and ceftazidime was done with and without the presence of clavulanic acid by agar dilution technique. Polymerase chain reaction revealed the presence of CTX-M type ESBLs in 19 isolates. Further sequencing resulted in identification of CTX-M-15 ESBLs. This is the first report identifying CTX-M type ESBL from clinical isolates of E. coli and K. pneumoniae from a tertiary care hospital in south India

Evaluation on the Use of β-Lactamase and Aminoglycoside Modifying Enzyme Gene Sequences as Markers for the Early Detection of Antibiotic Resistance Profile of Pseudomonas aeruginosa

Pseudomonas aeruginosa is one of the major causes of infections including the hospital acquired (Nosocomial) infections. Detection of them and their antibiotic resistance profile by conventional method takes about three days. Recently, DNA based diagnostic methods are being used for the identification of the pathogens. Hence we have tested a rapid and sensitive method using DNA sequences as markers for detecting the presence of three genes coding for the enzymes that inactivate the two most commonly used Anti-pseudomonadal drugs such as β-lactam antibiotics (Penicillin, and its derivatives) and Aminoglycosides such as Gentamicin, Tobramycin, Amikacin, Streptomycin. The internal region of these genes were used for designing and synthesizing primers and these primers were used in Polymerase Chain Reaction (PCR) to screen for the presence of these genes in the clinical isolates and to label them non-radioactively with Biotin. They in turn were used to detect the presence of the antibiotic resistance genes in the clinical isolates by hybridization. The specificity (ratio of positive results obtained in both methods and the sensitivity (the minimum amount of sample DNA and the labeled probe required for the tests) were evaluated

Microwave Assisted Synthesis of 1-[5-(Substituted Aryl)-1H-Pyrazol-3-yl]-3,5-Diphenyl-1H-1,2,4-Triazole as Antinociceptive and Antimicrobial Agents

Purpose: An efficient technique has been developed for microwave assisted synthesis of 1-[5-(substituted aryl)-1H-pyrazol-3-yl]-3,5-diphenyl-1H-1,2,4-triazole as antinociceptive and antimicrobial agents. Methods: The desired compounds (S1-S10) were synthesized by the microwave irradiation via cyclization of formerly synthesized chalcones of 3,5-diphenyl-1H-1,2,4-triazole and hydrazine hydrate in mild acidic condition. All newly synthesized compounds were subjected to study their antinociceptive and antimicrobial activity. The analgesic potential of compounds was tested by acetic acid induced writhing response and hot plate method. The MIC values for antimicrobial activity were premeditated by liquid broth method. Results: The compounds S1, S2, S4, S6 and S10 were found to be excellent peripherally acting analgesic agents when tested on mice by acetic acid induced writhing method and compounds S3, S6 and S1 at dose level of 100 mg/kg were exhibited superior centrally acting antinociceptive activity when tested by Eddy’s hot plate method. In antimicrobial activity compound S10 found to be broad spectrum antibacterial agent at MIC value of 15.62 μg/ml and compound S6 was exhibited antifungal potential at 15.62 μg/mL on both fungal strains. Conclusion: Some novel pyrazoles clubbed with 1,2,4-triazole derivatives were synthesized and evaluated as possible antimicrobial, centrally and peripherally acting analgesics

Polymeric ocular hydrogels and ophthalmic inserts for controlled release of timolol maleate

Background: Ophthalmic drug delivery systems are the challenging subject for the researchers because of delicate nature of ocular membrane and preventive barriers leading to less than 1 % of Bioavailability. Reasons for reduced bioavailability are due to rapid pre corneal elimination, tear turnover, lacrimal drainage, blinking and degradation by enzymes. Less bioavailability causes short duration of action and increased frequency of administration. Materials and Methods: Timolol maleate was used as model drug. Dynamic drug release studies were used to study the polymeric hydrogels and ophthalmic inserts. Rheological studies were carried out by Brookfield Viscometer LVDV- II+. Result and Discussion: Viscosity value lies in the range of 4.08 to 31.8 cps. Drug release data was fitted to various kinetic equations such as First order plots, Higuchi plots, Peppa′s exponential plots. The results shows fairly linear curve and the slope value of the Peppa′s equation is less than 0.5 and hence follows the fickian diffusion. Conclusion: The developed hydrogels and inserts were therapeutically effacious, stable, non irritant and provide a sustained release of drug over 8 hours time period

Stimuli-sensitive hydrogels: A novel ophthalmic drug delivery system

<b>Background:</b> Stimuli-sensitive hydrogels are three-dimensional, hydrophilic, polymeric networks capable of imbibing large amounts of water or biological fluids on stimulation, such as pH, temperature and ionic change. <b>Aim:</b> To develop hydrogels that are sensitive to stimuli, i.e. pH, in the cul-de-sac of the eye for providing a prolonged effect and increased bioavailability with reduction in frequency of administration. <b>Materials and Methods:</b> Hydrogels were formulated by using timolol maleate as the model drug, polyacrylic acid as the gelling agents, hydroxyl ethyl cellulose as the viscolizer and sodium chloride as the isotonic agent. Stirring of ingredients in pH 4 phosphate buffer at high speed was carried out. The dynamic dialysis technique was used for drug release studies. <i>In vivo</i> study for reduction in intraocular pressure was carried out by using albino rabbits. <b>Statistical Analysis:</b> Drug release studies data were used for statistical analysis in first-order plots, Higuchi plots and Peppas exponential plots. Student t-test was performed for <i>in vivo</i> study. <b>Results:</b> Viscosity of the hydrogel increases from 3.84 cps to 9.54 cps due to change in pH 4 to pH 7.4. The slope value of the Peppas equation was found to be 0.3081, 0.3743 and 0.2964. Up to 80&#x0025; of drug was released in an 8 h drug release study. Sterile hydrogels with no ocular irritation were obtained. <b>Conclusions:</b> Hydrogels show increase in viscosity due to change in pH. Hydrogels were therapeutically effacious, stable, non-irritant and showed Fickian diffusion. <i>In vivo</i> results clearly show a prolonged reduction in intraocular pressure, which was helpful for reduction in the frequency of administration

Achieving asepsis of banana leaves for the management of toxic epidermal necrolysis

Background: Banana leaf is used in many centers in India during the care of patients with toxic epidermal necrolysis (TEN) and other extensive blistering disorders. Sepsis is an important cause of death in TEN patients and use of banana leaf may be a source of such infection. Aims: We conducted this study to detect the bacterial flora of the banana leaf and to examine various methods of rendering the leaf aseptic. Methods: Five pieces of banana leaf, 2 x 2 cm in size, were cultured separately in blood agar as follows: One piece was heated over a flame and one was soaked in boiling water and one was autoclaved. Methylated spirit was applied over one piece and ignited. One piece was placed on the media, 'as is.' The Petri dishes were incubated examined after 48 h. Results: All the pieces except the autoclaved specimen of the leaf grew coagulase-negative staphylococci (CONS) when aseptic precautions were not maintained and aerobic spore bearers when all aseptic measures were subsequently instituted during the procedure. Conclusion: We recommend measures to prevent possible transmission of bacterial infection by the leaf. Autoclaved and aseptically handled banana leaves may be used to reduce chance of infection in the treatment of TEN