HPLC

A new, simple, specific, accurate and precise RP-HPLC method was developed for determination of Baclofen. In the present study, stress testing of Baclofen was carried out according to ICH guidelines Q1A (R2). Baclofen was subjected to stress conditions of hydrolysis, oxidation, photolysis and neutral decomposition. Extensive degradation was found to occur in acidic, condition. Mild degradation was observed in basic and at thermal conditions. Successful separation of drug from degradation products formed under stress conditions was achieved on a Hypersil BDS C18 column (250 mm × 4.6 mm, 5.0 μ particle size) using acetonitrile: acetate buffer (pH 3.7 ± 0.05) (50:50 v/v), at a flow rate of 1.0 mL/min and column was maintained at 40˚C. Quantification and linearity was achieved at 272 nm over the concentration range of 5 - 100 μg/mL for Baclofen. The method was validated for specificity, linearity, accuracy, precision, LOD, LOQ and robustness.

A simple, selective, linear, precise and accurate HPLC method was developed and validated for rapid assay of Oxazepam in Bulk and Pharmaceutical tablet Formulation. Isocratic elution at a flow rate of 1.0 ml/min was employed.  The chromatographic analysis was performed on a ODS 5 μm (4.6 mm x 15 cm) or equivalent column at ambient temperature. The mobile phase consisted of Methanol: Water in the ratio of 65:35v/v. The UV detection wavelength was 230nm and 100µl sample was injected. Flow rate was found to be 1.0.  The retention time for Oxazepam was identified. The Average percentage recovery of the method was in the range of 0.5.  The method was validated as per the ICH guidelines. The method was successfully applied for routine quality control analysis of pharmaceutical formulation.

A reverse phase HPLC method is developed for the determination of Azilsartan medoximil and chlorthalidone in pharmaceutical dosage forms. Chromatography was carried out on a C18 column 4.6 x 100mm, 5µm, Make: BDS using a mixture of potassium Di hydrogen ortho phosphate buffer and acetonitrile (35:65 v/v) as the mobile phase at a flow rate of 1.0 ml/min. Detection was carried out at 273 nm . The retention time of Azilsartan Medoxomil and Chlorthalidone was 2.59±0.1 mins and 3.85±0.5 min respectively. The linearity was observed In range of 2.5-15 µg/ml and 10-60 µg/ml with a correlation coefficient of Azilsartan medoximil and chlorthalidone were 0.996 and 0.999.the proposed method was validated for its linearity, accuracy, precision and robustness. The proposed method was found to be simple, rapid, accurate and precise. It was found to be economical and suitable for simultaneous determination of Azilsartan Medoxomil and Chlorthalidone in pharmaceutical dosage form.

The present study describes screening for antimicrobial and anthelmintic activity of the petroleum ether and alcoholic extracts of Corchorus depressus Linn. (Tiliaceae). The plant has been used in the traditional Indian system of medicine as an antibacterial, antifungal and anthelmintic drug, as a tonic, cooling medicine in fevers; its mucilage is prescribed in gonorrhea. Root is rubbed on stone and smeared over forehead to get relief in migraine; extract of plant is applied as a paste in healing of wounds. The HPLC chromatograms of the methanolic extract shows a single large peak with a retention time 6.451 min along with other components.  The Anthelmintic activity was studied on adult Indian earthworms, ‘Pheretima posithuma’. The methanolic extract produced more significant Anthelmintic activity than petroleum ether extract and the activities are comparable with the reference drug Piperazine citrate. The methanolic whole plant extract exhibited significant anti bacterial, antifungal activity, comparable to the standard drug tetracycline. The present study confirms the folklore claim of the plant utilized for its antimicrobial activity

A rapid, sensitive, precise, and accurate HPLC method coupled with a photodiode array detector was developed, optimized, and validated for the estimation of everolimus in bulk and tablet dosage forms. The chromatographic separation was achieved using a kromosil C8 column (200 mm × 4.6 mm i.d., 5 μm particle size) at 30°C temperature. The isocratic mobile phase consisted of 0.1M dipotassium hydrogen phosphate and methanol (60:40, v/v). The mobile phase was delivered at 1.0 ml/min and the analyte was monitored at 277 nm. The method was successfully validated in accordance to International Conference on Harmonization. The retention time of everolimus was found to be 3.496 min, and the calibration curve was linear in the concentration range of 50-150 μg/ml (R2 = 0.9992). The limit of detection and the limit of quantitation were found to be 0.109 μg/ml and 0.364 μg/ml, respectively.  The proposed method is accurate (percent recoveries were in the range of 99.56-100.09%) and precise (percent relative standard deviation - 0.047 %). No chromatographic interferences from the tablet excipients and components of mobile phase were found. The proposed method was found to be suitable for the quantitative determination of everolimus in tablet dosage forms.

A simple, precise, and rapid high performance liquid chromatography (HPLC) method for the determination of cefazolin level in human plasma using ceftriaxone as an internal standard (IS) was developed and validated. 0.25ml plasma samples containing cefazolin were mixed with 17.5 µg of the IS. After adding 0.3 ml of cold methanol kept in fridge at 4 °C, the mixture was vortexed for two min and then centrifuged for 15 min at 16000 rpm at room temperature. The clear supernatant was transferred into an auto-sampler vial and 100 µl was injected into the HPLC system with a run time of 18.0 min. The compounds of interest were efficiently separated on 4.6 x 150 mm Atlantis dC18-5µm steel column, using a Guard Pak pre-column module with Nova-Pak C185-µm insert, and detected using Waters 2998 photodiode array detector set at 270 nm. The mobile phase consisted of a mixture of 0.02 M of cetyltriethyl ammonium bromide and 0.01 M dipotassium hydrogen phosphate (pH = 6.5, adjusted with phosphoric acid) and acetonitrile, 60:40 (v:v) for 6 min, 50:50 for 6 min, and 60:40 for 6 min. It was delivered at a flow rate of 1.5 ml/min. No interference in blank plasma or by commonly used drugs was observed; and the detection limit of cefazolin was 0.1 µg/ml. The relationship between cefazolin concentration in plasma and peak area ratio of cefazolin/IS was linear (r2 ≥ 0.9979) in the range of 0.2–200 µg/ml. Intra- and inter-day coefficient of variation (CV) was ≤ 7.4%  and  ≤ 5.7%, with the corresponding bias of ≤ 4.9% and ≤ 5.1%. Mean extraction recovery of cefazolin and IS were 97%, respectively. Using the method, cefazolin was found to be stable in processed samples for 48 hrs at -20°C ( ≥ 96%), and in unprocessed samples for 8 weeks at -20°C (≥ 98%), Further, the method was successfully used to measure cefazolin level in plasma samples.

An accurate, simple, rapid and sensitive HPLC method has been developed for the determination of mebhydroline napadisylate in the tablet. The Chromatography was performed on a reversed phase C-18 column(150 mm × 4.6 mm id, 5μm) by isocratic elution, using a mobile phase of acetonitrile : ammonia 25% (80 : 20 v/v) at ambient temperature 25±5 °C and UV detection operates at 320 nm in an overall analysis time of about 10 min.The total retention time was 1.612 min with a flow rate of 1.0 ml/min. % 0f RSD values for precision is found to be 0.293 (< 2).The limits of detection (LOD) and quantification (LOQ) were 0.03µg/ml and 0.096133µg/ml, respectively. The correlation coefficient for Mebhydroline Napadisylate 0.9972 indicates linearity of the methods within the limits. The linear range of determination for Mebhydroline Napadisylate was 100-500μg/ml. However, the change in flow rate and column temperature also did not show any significant variance. The % recovery was found to be 99.70%-99.41%.  As per ICH guidelines the proposed method is fully validated and found to be linear over a workable drug concentration, accurate, precise and robust.  This HPLC method is selective, precise, and accurate and can be used for routine analysis of pharmaceutical preparation in industrial quality-control laboratories.

The analysis of improved HPLC-UV detector method for the separation and quantification of Acetaminophen, Dextromethorphan hydrobromide and Doxylamine succinate is described. Samples are analysed by means of reverse phase (RP-HPLC) using a Zodiac C-18, (250 × 4.6 mm, 5 µ), and the mobile phase consists of two Channels A and B. Channel-A pH 6.0 Buffer: Methanol (85:15) and Channel-B pH 3.0 Buffer: Acetonitrile (70:30). The flow rate is 1.0 ml/min. The column temperature was maintained at 30˚C and sample temperature was maintained at ambient and wavelength fixed at 245nm UV-detection. It is found that the method of RP-HPLC with UV-detection system for the analysis of Acetaminophen, Dextromethorphan hydrobromide and Doxylamine succinate is straight forward and applied in qualitative and quantitative analysis. The developed LC method was validated with respect to specificity, precision, linearity, ruggedness, stability of analytical solution and robustness. Validation study compared as per ICH guideline.

Compendia methods from the USP (United States pharmacopoeia) are widely used in Pharmaceutical drug product testing. However USP methods are under uninterrupted revision to improvement. Donepezil USP monograph is having two methods to quantify donepezil Related substance. The present research work is a single, simple and economic stability indicating impurity profiling method has been developed for scrutiny of Donepezil hydrochloride. Successfully Chromatographic separation has been achieved on an Inertsil C8 3v (150mm x 4.6mm) 3μm with buffered mobile phase consisting of solvent A (mixture of 0.1M phosphate (pH 2.8) buffer and methanol in the ratio 90: 10 (v/v); respectively) and Solvent B (mixture of 0.1 molar (M) phosphate ( pH 2.8) buffer , Acetonitrile and methanol in the ratio 20:20: 60 (v/v); respectively) delivered at flow rate of 1.0 mL/min and the detection wavelength is 215 nm. The drug was subjected to the stress conditions. Donepezil hydrochloride was found to deteriorate significantly in basic, oxidative stress conditions and stable other degradation conditions. One degradant was observed in the stability studies, Which is crossing Identification threshold .the same was isolated and structural elucidation was carried out by H-NMR, Mass spectroscopy. The developed method was corroborated as per ICH guidelines.

A reverse phase HPLC method is developed for the determination of Esomeprazole and Domperidone in pharmaceutical dosage forms. Chromatography was carried out on a C18 column [4.6 x 100mm, 5mm, Make: BDS] using a mixture of potassium di hydrogen ortho phosphate buffer and acetonitrile (65:35 v/v) as the mobile phase at a flow rate of 1.3 ml/min. Detection was carried out at 298 nm .The retention time of Domperidone and Esomeprazole was 2.788 min and 3.485 min. The linearity was observed In range of 50-130 µg/ml and 60-140 µg/ml with a correlation coefficient of Domperidone and Esomeprazole were 0.999 and 0.999.the proposed method was validated for its linearity, accuracy, precision and robustness .The proposed method is simple, accurate, precise, and reproducible hence it can be applied for routine quality control analysis of Esomeprazole and Domperidone in pharmaceutical dosage form.