HPLC

A simple, precise and sensitive reverse-phase high performance liquid chromatographic method was developed and validated for the simultaneous estimation of Ofloxacin and Flavoxate Hcl in pharmaceutical formulations. Chromatographic separation was performed on a High performance liquid chromatography equipped with auto sampler and UV detector. Good sensitivity for all analyte was observed with UV detection at wavelength of 301 nm, Separation was performed on a BDS Hypersil C18 (250 X 4.6mm) 5µm,using a mixture of 0.1% Triethyl Amine buffer pH 4 and Acetonitrile in the ratio of (40:60, v/v). The method results in excellent separation with good resolution between the two analytes. The within day variation between Ofloxacin and Flavoxate Hcl 1.72 and 1.97 % . The recovery was greater than 95 % with RSD less than 1.95 %. The method was validated according to ICH guidelines by performing linearity, accuracy, precision, limits of quantitation and selectivity. The results show the method is suitable for its intended use.

A simple, economic, selective, precise, and accurate High Performance liquid Chromatographic method for the analysis of Furazolidone in bulk drug and pharmaceutical formulations was developed and validated in the present study. The mobile phase consists of a mixture of potassium dihydrogen phosphate and acetonitrile 80:20. And Adjust pH 6.50±0.1 with dilute potassium hydroxide solution. This was found to give a sharp peak of Furazolidone at a retention time of 4.245 min. HPLC analysis of Furazolidone was carried out at a wavelength of 354 nm with a flow rate of 1.0 mL/min. The linear regression analysis data for the calibration curve showed a good linear relationship with a regression coefficient (r2) of 1 in the concentration range of 25 to 150 µg ml-1. The linear regression equation was y =46002x+40407. The developed method was employed with a high degree of precision and accuracy for the analysis of Furazolidone. The developed method was validated for accuracy, precision, robustness, detection and quantification limits as per the ICH guidelines.  The wide linearity range, accuracy, sensitivity, short retention time and composition of the mobile phase indicated that this method is better for the quantification of Furazolidone.

A simple, specific, sensitive, precise and stability-indicating reversed phase high performance liquid chromatographic method was developed and validated for the simultaneous determination of thiocolchicoside and aceclofenac, using a RP-18 column and a mobile phase composed of 0.1% trifluoroacetic acid: acetonitrile (45:55). The retention time of thiocolchicoside and aceclofenac were found to be 2.35 and 13.2 min, respectively. Linearity was established for both thiocolchicoside and aceclofenac in the range of 0.08-0.8 and 1-10 µg/ml. Both the drugs were subjected to acid, alkali and neutral hydrolysis, oxidation, dry heat, and photolytic degradation. The degradation studies indicated that both thiocolchicoside and aceclofenac were susceptible to acid, alkaline and neutral hydrolysis. The degradation products of thiocolchicoside and aceclofenac were well resolved from the pure drugs with significant differences in the retention time values. This method can be successfully employed for simultaneous quantitative analysis of thiocolchicoside and aceclofenac in bulk drugs and formulations.

Lawsonia inermis (Lythraceae) commonly called as Henna or Mailanchi is known for its cosmetic properties. Henna is an important source of phytochemicals of immense medicinal and pharmaceutical significance such as alkaloids, naphthoquinone derivatives, aliphatic components, tannins, phenolic derivatives, coumarins, xanthones, flavanoids, therefore an effort to compile all the information reported on its phytochemical and pharmacological activities, so that interest could be diverted towards this dye herb, for the treatment and relief from various ailments and diseases. The resultant extracts were analyzed by thin layer chromatography (TLC), column chromatography, ultraviolet spectroscopy (UV), Infrared (IR) spectroscopy, and High performance liquid chromatography (HPLC) techniques UV analysis was done by diluting the extracts with their respective solvents. 2 to 3 peaks were observed for extracted samples at 212 nm, 238.60 nm and 283.30 nm in methanol extract, 241 nm in chloroform extract, 238 nm in chloroform: methanol extract and 272.20 nm in ethyl acetate extract respectively. In HPLC  compounds  methanol extract is carried out for HPLC separation, and mainly three peaks were observed at 2.057, 2.522, 4.133 min retention times, which is the confirmation of presence of three separated compounds.

An accurate and precise high performance liquid chromatographic method was developed for quantitative estimation of bosentan in tablet dosage forms. Chromatographic separation of the drug was achieved on a Kromasil C18 column (150 x 4.6 mm; 5µ) by eluting with a mobile phase consisting of phosphate buffer (pH 4.0) and acetonitrile (30:70 v/v) at a flow rate of 1.0 mL/min. The drug in the eluate was monitored by U V detection at 270 nm. The retention time obtained for the drug was 3.54 min. The calibration curve plotted was linear over the range of 25-175 µg/mL of the drug. The validation of the method was done following the ICH guidelines. The proposed method could be applied for determination of bosentan in its tablet dosage forms without any interference from excipients. The method is suitable for routine quality control analysis of bosentan formulations.

An accurate, precise and reproducible high performance liquid chromatographic method was developed for quantitative estimation of emtricitabine, efavirenz and tenofovir disoproxil fumarate simultaneously in tablet dosage forms. Separation of the drugs was achieved within 15.0 min on a Xterra RP-18 column (150 x 4.6 mm; 5µ) by gradient elution using mixtures of ammonium acetate buffer and acetonitrile as the mobile phase. The analytes in the eluate were monitored at 260 nm. The retention times obtained for emtricitabine, efavirenz and tenofovir disoproxil fumarate were 4.611, 9.098 and 7.512 min respectively. The calibration curves were linear over the range of 20.11-60.33 µg/mL for emtricitabine, 60.28-180.45 µg/mL for efavirenz and 30.13-90.18 µg/mL for tenofovir disoproxil fumarate. The performance of the method was validated according to ICH guidelines. The method was found to be suitable for accurate determination of these drugs in tablet dosage forms without any interference from excipients or endogenous substances.

The neem tree has been known for its unique properties both in improving human health and against insects. Azadirachtin is a tri terpenoid limonoid obtained from various parts of the neem. Phosphatidyl choline is the most important phospholipid which helps in crossing the lipid barrier of the cell membrane and thus helps in easy absorption of the compound/drug. In the present work we are combining the isolated Azadirachtin and PC to form a complex, which can enhance the bio availability and easy absorption of the compound/drug. First of all, Azadirachtin is extracted from the dried neem kernel powder in a Soxhlet extractor using Di-Chloro methane as a solvent. Phosphatidyl choline is extracted and isolated from soya lecithin oil through reflux, using Methanol + Sodium hydroxide as solvent. The TLC, UV, HPLC, FTIR and DSC reports indicate the separation of the compounds from their crude forms. Azadirachtin and Phosphatidyl choline separated are refluxed together for 2 hours and dried in a Rota evaporator to form a 'Complex'. The absorbance's of Azadirachtin, Phosphatidyl choline and the Complex are measured at 220, 230 and 217nm. The qualitative analysis of Azadirachtin, Phosphatidyl choline and the Complex (1:1) were carried out by HPLC, on a C-18 column. Aceto nitryl: Methanol: 1% Triethyl amine pH 4 (60:40:1) was used as mobile phase at a flow rate of 1ml/min, at 210nm.The isolation of Phosphatidyl choline from soya lecithin oil used is a new method and the formation of Complex is an innovative work.

An accurate, precise and reproducible high performance liquid chromatographic method was developed for the simultaneous estimation of lamivudine, zidovudine and nevirapine in pharmaceutical dosage forms. Phenomenex C18 column (250 x 4.6 mm; 5µ) was employed for the separation of drugs. A mixture of 0.02 M trichloroacetic acid (6.8 pH) and methanol in the ratio of 40:60 v/v was used as the mobile phase and pumped at a flow rate of 1ml/min. The detection wavelength was set at 265 nm. The linearity of quantification was observed in the range of 7.5-112.5, 10-150 and 15-225 μg/ml for lamivudine, zidovudine and nevirapine respectively. The proposed method was validated according to ICH guidelines. The method was found to be suitable for simultaneous and accurate determination of these drugs in tablet dosage forms without any interference from the excipients.

A simple, selective, rapid, precise and economical reverse phase HPLC method has been developed for the determination of Metoprolol Succinate in dosage formulation. The analyte was resolved by using a mobile phase (Acetonitrile, water and 1 % ortho phosphoric acid in the ratio 70:27:3 v/v/v) at a flow rate 2.0 ml/min on an isocratic HPLC system (Agilent 1100 series with Chemstation software) consisting UV lamp detector, Aligent C-8, RP column (4.6 mm i.d x250 mm) at a wavelength of 280 nm.  The linear dynamic range for Metoprolol Succinate was 10 g/mL–200µg/mL. The limit of detection [LOD]and Limit of quantification[LOQ] for Metoprolol Succinate  was 0.0284µg/mL  and 0.094µg/mL respectively. 

An accurate, precise and reproducible high performance liquid chromatographic method was developed for quantitative estimation of abacavir, lamivudine and zidovudine simultaneously in tablet dosage forms. Separation of the drugs was achieved within 15.0 min on a Hichrom RP-select B column (250 x 4.6 mm; 5µ) by gradient elution using mixtures of 0.02M ammonium acetate and methanol as the mobile phase. The analytes in the eluate were monitored at 250 nm. The retention times obtained for abacavir, lamivudine and zidovudine were 12.172, 1.884 and 4.378 min respectively. The calibration curves were linear over the range of 25-200 µg/mL for abacavir, 12.5-100 µg/mL for lamivudine and 25-200 µg/mL for zidovudine. The performance of the method was validated according to ICH guidelines. The method was found to be suitable for accurate determination of these drugs in tablet dosage forms without any interference from the excipients or endogenous substances. Key words: Abacavir, Lamivudine, Zidovudine, Determination, HPLC, Gradient elution.