Development and Quality evaluation of sustained release pellets of eperisone HCl.

The objective was to develop eperisone HCl sustained-release pellets through extrusion spheronization technique and to determine the influence of different hydrophobic (polymeric based and wax-based) and hydrophilic (polymeric based) matrix former on the release of eperisone HCl (BCS class I drug) and on pellet sphericity. The pellet formulations consisted of different hydrophobic and hydrophilic matrix formers like HPMC K4M (10-20%) HPMC K15M (10%), EC (7cps) (10-20%), Carnauba wax (10-20%), Compritol ATO 888 (10-20%), Glyceryl monostearate (10%), lactose and microcrystalline cellulose. The initial burst release of the drug from matrix pellet formulations was effectively controlled by coating with 5% EC (ethylcellulose) dispersion. The dissolution profile and drug release kinetics of coated pellet formulations were determined at both acidic and basic pH medium. SEM (Scanning electron microscope) technique was used to determine the surface morphology and cross-section of F5 and F7 pellet formulation. The mechanism of drug release of coated formulation followed non-Fickian diffusion. FTIR spectroscopy was conducted and no drug and excipients interaction was observed. The results had shown that optimized coated formulation was F5 and F7 which effectively extend the drug release for 12 hours.

Development and pharmaceutical evaluation of oral fast dissolving thin film of escitalopram: A patient friendly dosage form.

The current study focused on the development, optimization and pharmaceutical evaluation of a mouth dissolving film of Escitalopram 5mg. The designing and optimization of the formulations have been carried out through Design-Expert ⌖ Ver. 9, using central composite response surface methodology. The software generated six optimized formulations have been fabricated via solvent casting method incorporated with HPMC and PEG 400 as Plasticizer. The developed formulations were assessed for various quality attributes including weight variation, drug-excipients interaction, dryness/ tack test, thickness, percent elongation, swelling index, disintegration, folding endurance, surface pH, content uniformity, assay, moisture uptake, stability, and organoleptic properties. A validated spectrophotometric method has been used to ascertain drug content. The formulations were subjected for stability studies for six months in accordance with ICH accelerated stability studies guidelines. No stability issue has been observed. All the test formulations have shown satisfactory in vitro release of escitalopram whereas most promising results have been exhibited by F5 and F6 formulations. The study concluded that a unique, novel, safe and stable formulation of oral fast dissolving thin films of escitalopram can be formulated with ease. The preparation method was simple and reproducible with scale-up tendency so that it can fulfill the need of the commercial manufacturing process.

High prevalence of DR-TB (drug-resistant tuberculosis): An Indicator of public health negligence.

Tuberculosis (TB) is among the 10 most common worldwide causes of mortality. In Pakistan, estimated 510,000 tuberculosis patients had been diagnosed with an occurrence of 276/100,000. As per most recent global TB report 2018, Pakistan is amongst the 30 countries high TB with drug-resistant Mycobacterium tuberculosis particularly MDR (multi-drug resistant strains). A retrospective study had been designed using DR-TB patients' records from January 2013 to the December 2017 year from a public sector hospital in Karachi. Overall 315 drug-resistant tuberculosis patient's data had been incorporated in the study. All data had been analyzed using SPSS version 16 software. Chi-square test had been used to analyze the data with CI (confidence interval) 95% and level of significance 5%. The study result showed that 64.1% MDR patients, 27.9% MTB rifampicin resistance, 4.8% mono-drug resistant , XDR(1.6%), 1% poly-drug resistant and only 0.6% are MDR suspects showing no association of DR-TB with gender (p-value 0.787), age group (p-value 0.757), treatment outcomes (p-value 0.549), year of registration( p-value 0.206), first line treatment history(p-value 0.643) with a 95% confidence interval. The drug resistance TB cases have been periodically rising every year. Early identification is required to reduce the percent mortality and inhibit the disease transmission.

Community onset of CTX-M extended spectrum ß-lactamases among uropathogenic E. coli and K. pneumoniae from Karachi, Pakistan.

Urinary tract infections (UTIs) are major health issue in developing countries like Pakistan, become more complicated with extended spectrum ß-lactamase (ESBL) expression in Escherichia coli and Klebsiella pneumoniae. The ground of this present study was to evaluate the incidence of cefotaxime (CTX-M) gene in Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis. The clinical isolates from various specimens were collected for one-year duration from January till December 2015. After initial screening (n=352) isolates were examined for phenotypic expression of ESBLs by double disc synergy test. Furthermore, eight-four isolates were analyzed by polymerase chain reaction for identification of Cefotaxime (CTX-M), Temoneira (TEM) and Sulfhdryl variable (SHV) genes. Among eighty-four clinical isolates CTX-M was dominant and found positive in 50 isolates (59.5%) followed by TEM in 35 (41.6%) and SHV in 11 (13%). In uropathogenic E. coli and K. pneumoniae, ESBLs gene was found in 50 and 6 isolates out of 57 and 7 respectively. Among uropathogens CTX-M was most prevalent 78% (39/50) in E. coli followed by K. pneumoniae. In uropathogenic E. coli, CTX-M was found dominant in females. The study concluded that ESBL related uropathogenic E. coli were CTX-M dominant, showed community onsets of UTIs that can be preventive and controlled with modified hygienic practices.

Pharmaceutical quality evaluation of different glimepiride brands marketed in Karachi (Pakistan): In pursuance to global issue of availability and affordability of quality medicines.

Availability of economical quality medicines is always required for chronic disease management. Price differences among multiple brands of a product do not essentially displays low quality for the more affordable brand, however in a few occurrences it appears. Glimepiride, an oral anti-diabetic drug, is produced by several national and multinational industries in Pakistan with considerable cost variation. The study aimed to evaluate the quality and economy of various Glimepiride brands available in Karachi, specifically of public sector hospitals. For this, eight glimepiride brands were collected and analyzed for the pharmaceutical quality using physical parameters, disintegration test, dissolution profile, spectrophotometric assay and content uniformity. Pharmacoeconomic assessment was also carried out such as availability, affordability and price variation. A profound discrepancy was observed among the prices of selected brands. All of the products found to be equivalent to the reference product except G5, the most inexpensive and highest consumed product of a public sector hospital. Study concludes that products with higher quality and lesser price can be used as a substitute to the costly brands while availability of a substandard product looks for consideration of pertinent authorities to assure the distribution of quality medicines.

Phenotypic expression and prevalence of multi drug resistant extended spectrum beta-lactamase producing Escherichia coli and Klebsiella pneumoniae in Karachi, Pakistan.

The aim of the present study was to evaluate the prevalence rate of ESBL producing Gram negative isolates of E. coli, K. pneumoniae and P. mirabilis, to determine the association of various factors with ESBL production and therapeutic options for the treatment. Total 352 isolates were subjected for identification of ESBL by double disc synergy test. Antimicrobial susceptibility was performed using CLSI guidelines and statistical association between ESBL/Non ESBL producers were determined by chi square at significant level of 0.05. A total of 96 isolates were ESBL positive (27%), females were 67% whereas males were 33%. E. coli was most prevalent pathogen (82%) followed by Klebsiella pneumoniae (17%). Furthermore 75% of ESBL associated infections were urinary tract infections. 95% of ESBL producing isolates were multidrug resistant and tazobactam/piperacillin combination and imipenem are good choices with 100% and 97% susceptibility respectively. E coli (OR 2.83, 95% CI 1.585-5.072, RR 2.22, p 0.0004) and K. pneumoniae (OR 0.52, 95% CI 0.285-0.952, RR 0.609, p 0.032) were significantly associated with ESBL production. The spread of ESBL producing multidrug resistant E. coli and K. pneumoniae has increased and proper screening for ESBL identification is needed because of limited therapeutic antibiotic choices.

Biowaiver studies of Metronidazole tablets (400mg): An alternative to In-vivo bioequivalence Studies.

The aim of the study was to investigate the dissolution behavior of commercially available brands of metronidazole and to provide basic tool to evaluate the comparative effectiveness and interchangeability of generic brands under biowaiver conditions. The dissolution test for six brands of metronidazole 400mg tablets was performed and physical controls were analyzed. Basket Rack methods at 100rpm were used to estimate release pattern of drug. Pharmaceutical parameters of tablets were analyzed. In order to evaluate dissolution profiles, multiple point dissolution were performed and calculated 85.96±0.41 to 90.56±0.93 % within 15 minutes in pH 1.2,85.50±1.40 to 88.99±0.80% in pH 4.5 and 85.37±1.94 to 92.79±0.89% in pH 6.8 dissolution medium respectively. Five different kinetics have been studied to predict and evaluate the acceptability level of drug release. The results show that Hixson-Crowell, first-order and Weibull demonstrated the drug release with R2> 0.95 that predicted the tablets were pharmaceutically equivalent. One-way ANOVA at p >0.05 level and similarity factors (f2) were used to estimate the discrepancy and intimacy among the brands. It is a need of time to constantly monitor the marketed generic drugs products and their release profiles to confirm their in vitro bioequivalence which can help to reduce the time, cost and unnecessary exposure of healthy subjects to medicines.

Some novel piperidine analogues having strong alpha glucosidase inhibition.

The idea of this study is based on the marvelous fact of nojirimycin and deoxy nojirimycin, naturally occurring from piperidine class and having their role as alpha glucosidase inhibitors. In the present work some hydroxyl piperidine analogues have been synthesized and analysed for their hypoglycemic effect through glucosidase inhibition owing to the structural resemblance with nojirimycin. The activity was done by spectral absorbance analysis using acarbose as standard. Two analogues (I & IV) were found to pose excellent activity having 87.4 and 54.7% inhibition respectively, hence strengthening the idea of studying piperidine analogiues as glucosidase inhibitors due to structural similarity with nojirimycin.

Prescribing pattern of angiotensin receptor blocker: A study of errors and drug-drug interactions.

Prescriptions comprising multi-drug therapy mostly illustrate the prescribing error. The phenomenon of error is bonded with human inaccuracy. The erroneous practice is observed in under developed countries like Pakistan, Bangladesh and also in developed ones. Consequently drug-drug interaction is one of the most common error associated with potentially serious adverse response even death. Accordingly the present study was conducted to assess the prevalence of prescribing errors and drug-drug interactions in out-patients receiving angiotensin receptor blockers. The study was done with population size one hundred fifty prescriptions obtained from different out-patient settings in Karachi. The prescriptions were screened for prescribing errors and risk factors for drug-drug interactions. Drug-drug interactions were recognized by Micromedex.2.0.Drug-Reax®database. The most common type of error was omission error. These errors were patient's age, weight and diagnosis found in 51.3%, 97.3% and 74% of prescriptions, respectively. The prevalence of drug-drug interaction was 38%. A total of 746 drugs were prescribed with an average of 5 drugs per prescription and 450 medication errors were detected. Majority of the interaction were moderate (19.33%), others were minor (14%) and major (6%) in severity. Patients who prescribed many drugs (more than 5 drugs in a while) had a higher risk of developing drug-drug interactions (OR=4.76; 95% CI=2.30-9.64; p=0.0001*).The study data reports the occurrence of prescribing errors in Karachi and also necessitate the need of clinical pharmacist's services in health care system. The step will help to minimize the risk factors by having the drug prescriptions reviewed by the pharmacists.

Development and validation of HPLC method for the determination of Candesartan in human plasma.

Candesartan (CAN), an ARB-blocker, antihypertensive, was analyzed in human plasma by a simple, accurate and precise RP-HPLC (reverse phase-High performance liquid chromatography assay method which was then validated for its accuracy, specificity and precision. The mobile phase has a constitution of acetone, diethylamine and distilled water, while Phosphoric acid was used to adjust the pH to 2.5±0.1. This mobile phase was run at 1.1ml/min and the fluorescence wavelength was set to 392 nm. A C-18 HPLC, column particle size (5 µm) Mediterranean Sea ® L x 1.D. 25cm x 4.6 mm (Supelcosil) , with auto sampler injection volume of 30µl ,an internal standard Valsartan was utilized for chromatographic detection. Candesartan took a retention time of 6±0.5 minutes. This method was validated by the parameters of selectivity, accuracy, precision, repeatability, reproducibility, recovery, linearity and stability. Candesartan's calibration curves were found to be linear in the range of 200ng/ml to 3.125ng/ml and the coefficient of determination (r2) was found to be 0.99. Analytical recovery obtained was above 88%. Hence, this method has been found to be useful for determining Candesartan in plasma.

Prevalence of mecA: Genotyping screening of community acquired-MRSA isolates in Karachi, Pakistan.

Among resistant nosocomial and community pathogens, MRSA has become the most serious pathogen, causing life threatening infections worldwide. In S.aureus, quick and exact recognition of methicillin (cefoxitin) resistance has become essential. The benchmark for MRSA identification among S.aureus is the detection of the mecA gene that causes the expression of protein (PBP2a) culpable for classic ß-lactam resistance. However, the utter reliance on amplification of mecA gene as a hallmark in confirmation of methicillin (cefoxitin) resistant S. aureus is the matter of distrust by some investigators. The current investigation designed to analyse the prevalence of mecA gene among phenotypically positive MRSA isolates using molecular method and to correlate its prevalence to conventional techniques. Furthermore, antimicrobial sensitivity of mecA positive staphylococci was determined by Kirby Baeuer method. For this purpose, 201 clinical staphylococcal specimens were recovered from various diagnostic laboratories in Karachi City, Pakistan. Phenotypic existence of methicillin resistance in S. aureus was observed to be 51.7%. In contrast, when organisms were subjected for amplification of mecA gene by PCR, mecA positive isolates were 36/104 (35%) MRSA isolates. Current work raise question towards the usefulness of molecular identification of mecA gene in confirmation of methicillin resistance without correlating with conventional methods. Therefore, it is essential to consider the other possible resistance mechanisms for ß-lactams that may interact with mecA gene in the development of methicillin resistance mechanism in Staphylococcus.

Development & validation of reversed phase HPLC method for quantification of water insoluble API.

In the present work a specific, accurate, precise, and reproducible UV-HPLC method was developed and validated for the analysis of Aceclofenac. This method involved elution of Aceclofenac in a mobile phase which is composed of buffer pH 6.8 (i.e. using 0.01N KH2PO4) and HPLC grade Acetonitrile (60:40). Separation of the analyte was achieved using HPLC isocratic pump attached to the UV-VIS detectorC18, guard column and C18 column. The injection volume was 20µL, detected at 274 nm; flow rate: 1mL/min. Standard calibration curve was measured and found linear from 0.1 to 40µg/ml. The validation parameters were measured according to FDA guidelines and successful results were obtained. The presented analytical method could be employed for pharmacokinetic studies.

Chromatographic method development and validation for the determination of valsartan in biological fluid.

A swift, precise and simple HPLC bioanalytical technique with UV detection was established and validated for quantitative estimation of valsartan in human plasma. The analyte was separated from plasma by protein precipitation with acetonitrile and chromatographically separated on Zorbax SB-C18 (5µm, 4.6mm × 15cm) column. The solvent mixture system consisting of acetonitrile, water and glacial acetic acid (40:59:1 v/v), was pumped using isocratic mode at 1mL/min flow rate. Samples' detection of drug was made spectrophotometrically at a wavelength of 264nm. The analyte response was instituted to be linear from 0.06 to 8µg/mL with a regression value of 0.999. The accuracy of the proposed method was ranged between 97.2-100.3% with 5% RSD. The analytical recovery (>95%) was consistently observed and satisfactory sample stability was also found at different environmental conditions. In conclusion the reported bio-analytical method is easy and robust that was successfully utilized in estimation of valsartan in a pharmacokinetic study.

MRSA: Prevalence and susceptibility pattern in health care setups of Karachi.

This assessment aims to determine the prevalence of methicillin resistance and multidrug resistance (MDR) among the clinical isolates of Staphylococcus aureus and antimicrobial susceptibility profile of methicillin resistant Staphylococcus aureus (MRSA) to the frequently prescribed antibiotics in Karachi. Isolates of MRSA, recovered from various clinical samples were included in this prospective, cross-sectional study from Jan 2015 to June 2017. Agar diffusion method was employed according to the protocols of Clinical Laboratory Standards Institute. Out of total 346 S. aureus strains, the frequency rate of MRSA was 52% (n = 180). MRSA infection was found higher among the age group 21-30 years i.e. 30% (n=54), followed by 20% (n=36) in 31-40 years. Frequency of MRSA percentage in male and female was and 70% and 30% respectively. MRSA was more frequently observed in blood 20% (n=36). MRSA showed high resistance (100%) to Oxacillin and Cefoxitin while 25% Vancomycin resistant S. aureus (VRSA) isolates and 25% Teicoplanin resistance were also reported. MRSA exhibited 16% resistance to Minocycline. It was concluded that MRSA pose a challenging threat to public health in Karachi. In addition, MDR should be periodically checked to avoid treatment failure.

Determination of Cefadroxil in Tablet/Capsule formulations by a validated Reverse Phase High Performance Liquid Chromatographic method.

An innovative, selective and rapid reversed phase High Performance Liquid Chromatographic (RP-HPLC) method for the analysis of cefadroxil in bulk material and oral solid dosage forms has been developed and validated. The chromatographic system consisted of Sil-20A auto sampler, LC-20A pump and SPD-20A UV/visible detector. The separation was achieved by C18 column at ambient temperature with a mobile phase consisting of methanol: Phosphate buffer (10: 90) at a flow rate of 1.5 ml/min. The method is reproducible, repeatable (%RSD for intra-day and inter-day ranged between 1.75-5.33% and 0.58-2.69%) and linear (R2=0.9935). The LOD and LOQ of the method were 0.5 and 1.0 µg/ml, respectively. The present RP-HPLC method was found to be sensitive, accurate, precise, rapid and cost effective that can be efficiently used in QC/QA laboratories for routine analysis of the raw materials as well as oral dosage formulations of cefadroxil.

Disintegrants combination: development and optimization of a cefadroxil fast disintegrating tablet.

Fast Disintegrating Tablets (FDTs) is a rapidly growing dosage form preferred for special population (pediatric, geriatric and psychotic patients). It is also developed with the aim of improving bioavailability and patient compliance. During the present study, cefadroxil fast disintegrating tablets formulations (n=9) were designed and optimized by central composite design with two independent variables (croscarmellose and crospovidone) using design expert® software. The effects of independent variables on formulation properties such as friability, hardness, in vitro dispersion and disintegration were assessed by drawing response surface graphs with design expert® software. Tablets were assessed for pharmacopeial and non-pharmacopeial parameters to ensure the quality of compressed tablets. Among all formulations, F3, F8 and F9 have shown better results. The formulation F9 containing 15mg croscarmellose and 33.075mg crospovidone showed good pharmacotechnical attributes as well as shelf life. F 9 showed improved dissolution with t90% of> 2 min and will lead to better bioavailability.

Incidence of medication error associated with the use of beta-blockers in Pakistan.

Medication errors (ME) are human errors, which are very frequent in cardiovascular patients and result in patient morbidity and mortality. This study was focused to evaluate the quality of prescriptions and to emphasize the placement of clinical pharmacist in health care team. This study was carried out in different outpatient settings of (in) Karachi, Pakistan. The study period was June 2011 till June 2012. Retrospective data was analyzed for the outpatients' prescription of beta blocker drugs. During the study, prescriptions (n=450) were collected from different outpatient settings of (in) Karachi, Pakistan. Prescription containing beta-blockers were analyzed for the essential elements to be mentioned in prescription. Drug-drug interactions were identified by the Micromedex.2.0 Drug-Reax database and severity of medication error was determined by NCCMERP Index. A total of 1627 medication errors were identified in 450 prescriptions. The most frequent error was not mentioning the patient's weight (95%), followed by missing diagnosis (79.4%) and drug-drug interactions (69.5%). Twenty-two prescriptions were placed in the most severe category I (4.88%). Average number of drugs per prescription was 4.76. Significant difference was observed (χ(2)=52.418, p<0.05) using SPSS 19 for those prescription orders having more than 5 drugs with Beta-blockers. This indicates that the errors in prescription such as drug-drug interactions, wrong dose etc. was significantly increased with the number of drugs per prescription. Results showed that medication errors are very frequent in prescription written in outpatient setting of various hospitals and clinics in Karachi. This shows that the irrational prescribing is a common practice in developing countries. Placement of skilled pharmacist in the health care system is the only solution for avoidance of these medication related problems.

Formulation development of intermediate release Nimesulide tablets by CCRD for IVIVC studies.

Simple and cost effective study consisting of three steps, comparison of micromeritic properties of different blends i.e. placebo without API and Nimesulide containing, Use of central composite design (CCRD) for intermediate release Nimesulide tablets and stability results of three selected Nimesulide tablet formulations which were calculated by using R Gui. Different concentrations of Avicel, hydroxypropyl methyl cellulose (HPMC) and magnesium stearate were used as variables in central composite design and two types blend i.e., with or without Nimesulide were selected for bulk density, tap density, percentage compressibility; angle of repose and Hausner's ratio. Blending rate constant was performed after applying the different mixing times like 3, 6, 9 and 12 minutes. Twenty intermediate release formulations were designed and three formulations were chosen for compression by direct compression method on the basis of compressibility index. Physicochemical properties and best release pattern in four hours in different dissolution medium were successfully measured. Relative densities, porosity of tablets were compared with tensile strength of tablet and weight variation, hardness, friability and dissolution was performed by simple experiments. Presence of Nimesulide in the bulk increased all micromeratic tests while 9 minutes was best mixing time. The hardness of NM containing tablets increased with the increase of relative density. The release pattern was further analyzed by model dependent i.e. zero order, first order and Higuchi, Korse-meyer and Pappas, Hixson Crowell and model independent kinetic model i.e., f2 value respectively. R Gui explained the F16 formulation shows the best result in stability studies with shelf life 72 months.

Predictors of Successful First-Line Helicobacter pylori Eradication with Fluoroquinolones in Pakistan: A Prospective Exploration of Demographic and Clinical Factors.

Growing antibiotic resistance complicates H. pylori eradication, posing a public health challenge. Inconclusive research on sociodemographic and clinical factors emphasizes the necessity for further investigations. Hence, this study aims to evaluate the correlation between demographic and clinical factors and the success rates of H. pylori eradication. A group of 162 H. pylori-positive patients were allocated randomly to receive either a ten-day moxifloxacin-based triple therapy or a levofloxacin-based sequential therapy. Eradication success was determined through the stool antigen test. Logistic regression analysis was utilized to figure out potential factors that contribute to H. pylori eradication success. Significantly higher H. pylori eradication rates were observed in the middle age group (COR: 3.671, p = 0.007), among females (p = 0.035), those with BMI ≥ 25 (COR: 2.011, p = 0.045), and non-smokers (COR: 2.718, p = 0.018). In multivariate analysis, age and smoking emerged as significant predictors (p < 0.05). Patients with comorbidities, excluding diabetes and hypertension (COR: 4.432, p = 0.019), dyspepsia (COR: 0.178, p < 0.001), and moxifloxacin triple therapy (COR: 0.194, p = 0.000), exhibited higher chances of eradication (p < 0.05). Further research is vital for tailored approaches to enhance eradication success.

Antimicrobial susceptibility testing of newer quinolones against gram positive and gram negative clinical isolates.

Antibiotic resistance development is an ongoing process associated with irrational antibiotic use. WHO recommends regular surveillance programs for monitoring of antibiotic resistance. The present study is a step in this direction. A total of 124 clinical isolates of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa were collected from different hospitals in Karachi. In vitro antimicrobial susceptibility studies were carried out by agar dilution method using newer quinolones that included Gatifloxacin and Levofloxacin. It was observed that 50% (n=30) isolates of Staphylococcus aureus were resistant to gatifloxacin. Gatifloxacin was more active against Pseudomonas aeruginosa (n=23) and showing complete susceptibility with MIC 1mg/L except for three very resistant strains that shown resistance at even higher concentrations. Escherichia coli (n=45) has shown 15.5% and Klebsiella pneumoniae (n=26) 34.61% resistance to gatifloxacin. Levofloxacin was more active against Staphylococcus aureus and Escherichia coli showing complete susceptibility at 0.5 mg /L concentration. Pseudomonas aeruginosa and Klebsiella pneumoniae were found to be resistant to Levofloxacin showing 36.36% and 23.08% resistance respectively. The study strongly recommends the adherence to the antibiotic policy and regular susceptibility testing to overcome the problem associated with antimicrobial resistance.