Application of experimental design for the optimization of forced degradation and development of a validated stability-indicating LC method for luliconazole in bulk and cream formulation

AbstractA stability-indicating LC method was developed and validated for the determination of luliconazole in bulk and cream formulation. Luliconazole was exposed to acid, alkali and water hydrolysis, oxidation effect by hydrogen peroxide, dry heat and photolytic conditions. Full factorial design was used during forced degradation experiments and the factors/combination of factors which were most likely to effect degradation of luliconazole under various conditions were identified and further were optimized using surface response curve. Drug was found to be stable under wet heat and dry heat conditions, but substantial degradation was observed under acid, alkali, oxidative and photolytic conditions. Drug and its degradation products were optimally resolved on HiQ sil C-18HS (250×4.6mm, 5μm) column with the mobile phase consisting of methanol and water (80:20, v/v) at a flow rate of 1mL/min, detection was performed at 296nm. The procedure was validated for specificity, linearity, accuracy and precision. There was no interference of excipients and degradation products in the determination of active pharmaceutical ingredient. The method was accurate and precise and the response was found to be linear in the range of 2–14μg/mL. The method was found to be simple and fast by making use of experimental design

Optimization of Forced Degradation Using Experimental Design and Development of a Stability-Indicating Liquid Chromatographic Assay Method for Rebamipide in Bulk and Tablet Dosage Form

A novel stability-indicating RP-HPLC assay method was developed and validated for quantitative determination of rebamipide in bulk and tablet dosage form. Rebamipide (drug and drug product) solutions were exposed to acid and alkali hydrolysis, thermal stress, oxidation by hydrogen peroxide and photodegradation. Experimental design has been used during forced degradation to determine significant factors responsible for degradation and to obtain optimal degradation conditions. In addition, acid and alkali hydrolysis was performed using a microwave oven. The chromatographic method employed the HiQ sil C-18HS (250 × 4.6 mm; 5 μm) column with mobile phase consisting of 0.02 M potassium phosphate (pH adjusted to 6.8) and methanol (40:60, v/v) and the detection was performed at 230 nm. The procedure was validated for specificity, linearity, accuracy, precision and robustness. There was no interference observed of excipients and degradation products in the determination of the active pharmaceutical ingredient. The method showed good accuracy and precision (intra and inter day) and the response was linear in a range from 0.5 to 5 μg mL−1. The method was found to be simple and fast with less trial and error experimentation by making use of experimental design. Also, it proved that microwave energy can be used to expedite hydrolysis of rebamipide

DEVELOPMENT AND VALIDATION OF A BIOANALYTICAL RP-HPLC METHOD FOR AZILSARTAN MEDOXOMIL WITH LIQUID-LIQUID EXTRACTION

Objective: There are many analytical methods available for estimation of Azilsartan medoxomil in biological samples and for pharmaceutical preparations. However, no specific RP-HPLC method with UV detection based on liquid-liquid extraction technique is available for estimation of Azilsartan medoxomil in human plasma. Methods: A simple, rapid and accurate RP-HPLC with UV detection method was developed and validated as per US-FDA guidelines for the estimation of Azilsartan medoxomil in spiked human plasma using liquid-liquid extraction technique. Results: Azilsartan medoxomil was well resolved from human plasma interference and internal standard (Aceclofenac) using C 18 (250 × 4.6 mm, 5 μ) column with methanol: 20 mm phosphate buffer (pH 3.0), (70: 30 %, v/v) as mobile phase, at a flow rate of 1 ml/min. The detection was performed at 249 nm. The calibration curve was found linear in the range of 500-16000 ng/ml. During calibration experiments, the heteroscedasticity was minimized by using weighted least square regression model with weighing factor 1/x². In accuracy and precision studies, intra-day and inter-day, % relative error was found between±15 and % RSD was less than 15 %. Stability experiments indicated that the drug remained stable after three freeze-thaw cycles. Conclusion: A simple, rapid and accurate RP-HPLC method with UV detection was developed and validated for estimation of Azilsartan medoxomil based on liquid-liquid extraction technique. The developed method meets the requirements of US-FDA guidelines. Also the developed method does not require expensive chemicals and solvents and does not involve complex instrumentation, hence it is economic.Â

Cytomorphometric Analysis of Oral Mucosa in Diabetic Patients in Bhopal Region an In-Situ Study

Background: Diabetes is a common endocrine metabolic disorder and prevalence is increasing worldwide. In condition like diabetes, premalignant lesions and iron deficiency anemia; oral exfoliative cytology may be more appropriate as the invasive techniques lose viability. Aim: The study was conducted to analyze the cytomorphometric changes in exfoliated cells of oral mucosa as an adjunct to diagnosis of diabetes. Method: Samples were collected from buccal mucosa and divided into 2 groups; 100 diabetic patients (study) and 100 healthy individuals (control). The smears were stained with rapid Papanicolaou stain (PAP). Nuclear area (NA), cytoplasmic area (CA) and cytoplasmic to nuclear ratio (CNR) were evaluated in 50 cells in each smear using Image analysis software (Magnus pro™) and research microscope (Lawrance & Mayo™). Results: Mean NA was significantly higher (p < 0.001) in study group whereas mean CA didn’t exhibit any statistically significant difference (p > 0.001). The mean CNR was significantly lower in study group (p < 0.001). Conclusion: Clinical observations and results suggested morphologic and functional alterations in oral epithelial cells in diabetic patients; detectable by microscopic and cytomorphometric analysis using exfoliative cytology and can be used in diagnosis. &nbsp

What does quality of care mean for maternal health providers from two vulnerable states of India? Case study of Bihar and Jharkhand

Background: Quality instillation has its own challenges, facilitators and barriers in various settings. This paper focuses on exploration of quality components related to practices, health system challenges and quality enablers from providers\u2019 perspectives with a focus on maternal health studied through a pilot research conducted in 2012\u20132013 in two states of India\u2014Bihar and Jharkhand\u2014with relatively poor indicators for maternal health. Methods: Qualitative data through in-depth interviews of 49 health providers purposively selected from various cadres of public health system in two districts each from Bihar and Jharkhand states was thematically analysed using MAXQDA Version 10. Results: Maternity management guidelines developed by the National Health Mission, India, were considered as a tool to learn instillation of quality in provision of health services in various selected health facilities. Infrastructure, human resources, equipments and materials, drugs, training capacity and health information systems were described as health system challenges by medical and paramedical health providers. On a positive note, the study findings simultaneously identified quality enablers such as appreciation of public-private partnerships, availability of clinical guidelines in the form of wall posters in health facilities, efforts to translate knowledge and evidence through practice and enthusiasm towards value of guidelines. Conclusions: Against the backdrop of quality initiatives in the country to foster United Health Care (UHC), frontline health providers\u2019 perspectives about quality and safety need to be considered and utilized. The provision of adequate health infrastructure, strong health management information system, introduction of evidence-based education and training with supportive supervision must constitute parallel efforts

Characterization of CDK(5) Inhibitor, 20-223 (aka CP668863) for Colorectal Cancer Therapy

Colorectal cancer (CRC) remains one of the leading causes of cancer related deaths in the United States. Currently, there are limited therapeutic options for patients suffering from CRC, none of which focus on the cell signaling mechanisms controlled by the popular kinase family, cyclin dependent kinases (CDKs). Here we evaluate a Pfizer developed compound, CP668863, that inhibits cyclin-dependent kinase 5 (CDK5) in neurodegenerative disorders. CDK5 has been implicated in a number of cancers, most recently as an oncogene in colorectal cancers. Our lab synthesized and characterized CP668863 – now called 20-223. In our established colorectal cancer xenograft model, 20-223 reduced tumor growth and tumor weight indicating its value as a potential anti-CRC agent. We subjected 20-223 to a series of cell-free and cell-based studies to understand the mechanism of its anti-tumor effects. In our hands, in vitro 20-223 is most potent against CDK2 and CDK5. The clinically used CDK inhibitor AT7519 and 20-223 share the aminopyrazole core and we used it to benchmark the 20-223 potency. In CDK5 and CDK2 kinase assays, 20-223 was ~3.5-fold and ~65.3-fold more potent than known clinically used CDK inhibitor, AT7519, respectively. Cell-based studies examining phosphorylation of downstream substrates revealed 20-223 inhibits the kinase activity of CDK5 and CDK2 in multiple CRC cell lines. Consistent with CDK5 inhibition, 20-223 inhibited migration of CRC cells in a wound-healing assay. Profiling a panel of CRC cell lines for growth inhibitory effects showed that 20-223 has nanomolar potency across multiple CRC cell lines and was on an average \u3e2-fold more potent than AT7519. Cell cycle analyses in CRC cells revealed that 20-223 phenocopied the effects associated with AT7519. Collectively, these findings suggest that 20-223 exerts anti-tumor effects against CRC by targeting CDK 2/5 and inducing cell cycle arrest. Our studies also indicate that 20-223 is a suitable lead compound for colorectal cancer therapy

Search for top squarks in the four-body decay mode with single lepton final states in proton-proton collisions at s \sqrt{s} = 13 TeV

A search for the pair production of the lightest supersymmetric partner of the top quark, the top squark (t∼1), is presented. The search targets the four-body decay of the t∼1, which is preferred when the mass difference between the top squark and the lightest supersymmetric particle is smaller than the mass of the W boson. This decay mode consists of a bottom quark, two other fermions, and the lightest neutralino (χ∼01), which is assumed to be the lightest supersymmetric particle. The data correspond to an integrated luminosity of 138 fb−1 of proton-proton collisions at a center-of-mass energy of 13 TeV collected by the CMS experiment at the CERN LHC. Events are selected using the presence of a high-momentum jet, an electron or muon with low transverse momentum, and a significant missing transverse momentum. The signal is selected based on a multivariate approach that is optimized for the difference between m(t∼1) and m(χ∼01). The contribution from leading background processes is estimated from data. No significant excess is observed above the expectation from standard model processes. The results of this search exclude top squarks at 95% confidence level for masses up to 480 and 700 GeV for m(t∼1) − m(χ∼01) = 10 and 80 GeV, respectively

Measurement of the Dependence of the Hadron Production Fraction Ratios fs/fuf_s / f_u and fd/fuf_d / f_u on BB Meson Kinematic Variables in Proton-Proton Collisions at s=13TeV\sqrt{s} = 13 TeV

The dependence of the ratio between the $B^0_s$ and $B^+$ hadron production fractions, $f_s/f_u$, on the transverse momentum $(p_T)$ and rapidity of the $B$ mesons is studied using the decay channels $B^0_s→J/ψϕ$ and $B^+→J/ψK^+$. The analysis uses a data sample of proton-proton collisions at a center-of-mass energy of 13 TeV, collected by the CMS experiment in 2018 and corresponding to an integrated luminosity of $61.6  fb^{−1}$. The $f_s/f_u$ ratio is observed to depend on the $B$ $p_T$ and to be consistent with becoming asymptotically constant at large $p_T$. No rapidity dependence is observed. The ratio of the $B^0$ to $B^+$ meson production fractions, $f_d/f_u$, is also measured, for the first time in proton-proton collisions, using the $B^0→J/ψK^{*0}$ decay channel. The result is found to be within 1 standard deviation of unity and independent of $pT$ and rapidity, as expected from isospin invariance