Development of Solid-Phase Extraction and HPLC Method for Simultaneous Estimation of Ilaprazole and Glimepiride in Rat Plasma: Application to Pharmacokinetic Studies.

A novel, simple and mass spectrometry (MS) compatible high-performance liquid chromatography (HPLC) method is reported for the simultaneous estimation of ilaprazole (ILA) and glimepiride (GLM) in rat plasma. The bio-analytical procedure involves extraction of ILA, GLM and internal standard (IS) from rat plasma with a solid-phase extraction (SPE) process. The chromatographic analysis was performed on Waters-600 system using an isocratic mobile phase comprising methanol:water (80:20 % v/v) with pH of water modified to three using formic acid at a flow rate of 1.0 mL/min and Kinetex C18 column maintained at 30 ± 1°C. The signals were monitored using a PDA detector set at 225 nm. IS, ILA and GLM eluted at 2.04, 4.7 and 7.4 min, respectively, and the total run time was 10 min. Method validation was performed as per US Food and Drug Administration guidelines and the results met the acceptance criteria. The calibration curve was linear over a concentration range of 10-600 ng/mL (r2 = 0.999). The intra- and inter-day precisions for ILA and GLM were (%RSD values) in the range of 1.52-9.74 and 1.52-11.76%, respectively, in rat plasma. The method was successfully applied in pharmacokinetic studies followed by oral administration of GLM and ILA in rats.

Evaluation of Wound Healing Activity of Ethanolic Extract of Pongamia pinnata Bark.

Present study evaluate wound healing activity of ethanolic extract of stem bark of Pongamia pinnata (PP). Evaluation of wound healing activity, 2 wound models were used I. e., incision and excision wounds were perform in this study on Albino wistar rats (150-200 g). The rats were been treated with 10% and 5% ointment base formulation at dose 15 µl/wound topically. The parameters studied were breaking strength in case of incision wounds, epithelization period and wound area in case of excision wound. The ethanolic extract treated group showed a significant (P < 0.01) reduction in the wound breaking strength in incision type of wound model and significant increase in epithelization period and reduction in percentage of wound area in excision type of wound model as compared to control group. Extract treated groups showed significant (P < 0.01) improvement in all the wound healing parameters of incision and excision wound models as compared to control. This study justify the traditional use of ethanolic extract of Pongamia pinnata stem bark shows wound healing property.

Evaluation of drug interaction of glimepiride with phosphodiesterase inhibitors type V in diabetic nephropathy.

The present study investigate the drug interaction of Glimepiride (GLIM) with Sildenafil Citrate (SIL) in Diabetic Nephropathy (DN) rats. Diabetes was induced in rats by administering Streptozotacin i. e. STZ (60 mg/kg). In present investigation GLIM (0.5 mg/kg, P.O.) and SIL (2.5 mg/kg, P.O.) were given for 6 weeks after the confirmation of DN. Pharmacodynamic and kinetic parameters were estimated after 1(st) dose and at the end of 6(th) week of drug administration. There was significant (p<0.001) increase in the bioavailability of GLIM in the presence of SIL in DN after the 1(st) dose of administration of the drug as compared to GLIM treated rat, whereas at the end of 6(th) week of drug administration, there were significant (p<0.0001) decrease in the bioavailability of GLIM+SIL treated DN rats compared to GLIM treated rats. There was significant (P<0.01) reduction of blood glucose level in GLIM+SIL treated group as compare to only GLIM treated group on 1(st) dose of drug administration but after continuous treatment for next 6 weeks, GLIM treated group showed significant hypoglycemia which was found to be reduced in GLIM+SIL treated group significantly. The present study concluded that GLIM+SIL treatment reduces the hypoglycemic condition which was there for GLIM treated DN rat.

Development of health supplement probiotic in relation to physical strength and biological activity in experimental animal.

A probiotic nutritive health supplement having a healthy nutritive value was formulated to enhance body function. A 90 days study proved the formulated health supplement to improve the body weight with a maintained normal level of cholesterol and triglycerides in blood which is needful for the proper cardiac function. The supplement also maintained the blood parameters to avoid any illness in the body. In the present study, the health supplement was able to maintain the Haemoglobin level, RBC Count, WBC Count, Platelet Count in Sprague Dawley rats. It is also found that the supplement maintained the cardiac rhythm in the stressful condition and increased in the time of swimming in the force swimming test which indicates an increase in the physical strength.

Study of stress degradation behavior of rasagiline mesylate under hydrolytic conditions by high performance liquid chromatography.

A precise and rapid reverse phase HPLC method has been developed for the estimation of Rasagiline Mesylate in bulk drug under stressed hydrolytic conditions according to ICH guidelines Q1(A)R2. The stressed samples of Rasagiline Mesylate were analyzed using Prontosil ODS column. The mobile phase consists of methanol and Phosphate buffer. The pH of the phosphate buffer was adjusted to 3.0 using orthophosphoric acid. The composition of mobile phase Methanol: Phosphate buffer (75:25 v/v) was kept constant up to 1 minute and then changed to 90:10 v/v in next 9 min by HPLC gradient programming. The flow rate was maintained at 1.0 ml/min and detection was carried out at 210 nm using 996 PDA detector. The method was validated in terms of linearity, accuracy, precision specificity and selectivity. The method was found to be linear in the range of 5-15 µg/ml with a correlation coefficient of 0.993.

Gradient HPLC-DAD determination of paracetamol, phenylephrine hydrochloride, cetirizine in tablet formulation.

Present work describes the development and validation of a simple and reliable high-performance liquid chromatography-diode array detection (HPLC-DAD) procedure for the analysis of phenylephrine hydrochloride (PHE), paracetamol (PAR) and cetirizine dihydrochloride (CET), in pharmaceutical mixture. The method was applied successfully on tablet dosage form. Effective chromatographic separation of PHE, PAR and CET was achieved using a Kinetex-C18 (4.6, 150 mm, 5 mm) column with gradient elution of the mobile phase composed of 10 mM phosphate buffer (pH 3.3) and acetonitrile. The elution was a 3 step gradient elution program step-1 started initially with 2% (by volume) acetonitrile and 98% phosphate buffer (pH 3.3) for first 2 min. In step-2 acetonitrile concentration changed linearly to 20% upto 12 min the analysis was concluded by step-3 changing acetonitrile to 2% upto 20 min. The reliability and analytical performance of the proposed HPLC procedure were statistically validated with respect to linearity, ranges, precision, accuracy, selectivity and robustness. Calibration curves were linear in the ranges 5-15, 250-750 and 2.5-7.5 µg/ml for PHE, PAR and CET, with correlation coefficients >0.9996. The validated HPLC method was applied to a pharmaceutical mixture of a marketed preparation tablet in which the analytes were successfully quantified with good recovery values with no interfering peaks from the excipents.

Development and validation of RP-HPLC method for sildenafil citrate in rat plasma-application to pharmacokinetic studies.

Sildenafil citrate (SIL) is used in the treatment of erectile dysfunction and other chronic disorders. For the pharmacokinetic investigation of SIL we developed a simple and sensitive method for the estimation of SIL in rat plasma by reverse phase high-performance liquid chromatography (RP-HPLC). The drug samples were extracted by liquid-liquid extraction with 300 µl of acetonitrile and 5 ml of diethyl ether. Chromatographic separation was achieved on C18 column using methanol:water (85:15 v/v) as mobile phase at a flow rate of 1 ml/min and UV detection at 230 nm. The retention time of SIL was found to be 4.0 min having a separation time less than 5 min. The developed method was validated for accuracy, precision, linearity and recovery. Linearity studies were found to be acceptable over the range of 0.1-6 µg/ml. The method was successfully applied for the analysis of rat plasma sample for the application in pharmacokinetic study, drug interaction, bioavailability and bioequivalence.

Development and validation of RP-HPLC method for simultaneous estimation of glimepiride and sildenafil citrate in rat plasma-application to pharmacokinetic studies.

A simple and sensitive method was developed for simultaneous estimation of Glimepiride (GLIM) and Sildenafil citrate (SIL) in rat Plasma by reverse phase high performance liquid chromatography (RP-HPLC). The drug samples were extracted by liquid-liquid extraction with 300 µl of acetonitrile and 5 ml of diethyl ether. Chromatographic separation was achieved on C18 column using methanol: water (85:15 v/v) as mobile phase at a flow rate of 1 ml/min and UV detection at 230 nm. The retention time of GLIM and SIL was found to be 2.5 and 4.0 min respectively with total run time of 7 min. The developed method was validated for accuracy, precision, linearity and recovery. The method was linear and found to be acceptable over the range of 100-12 000 ng/ml. The method was successfully applied for the analysis of rat plasma sample for application to pharmacokinetic.