Nilesh Jain

Evaluation of wound-healing activity of Acorus calamus Linn.

The aim of the present study was to assess the wound-healing activity of ethanolic extracts of Acorus calamus leaves. A wound was induced by an excision- and incision-based wound model in rats of either sex. The mature green leaves of A. calamus were collected and authenticated. Extractions of dried leaves were carried out with 80% ethanol in a soxhlet apparatus. For wound-healing activity, the extracts were applied topically once daily in conc. of 40% w/w and 20% w/w in the form of ointment and compared with a standard drug (povidion-iodine). The healing of the wound was assessed by the rate of wound closure, period of epithelialisation, tensile strength and weight of the granulation tissue, hydroxyproline content and histopathology of the granulation tissue. The ethanolic extract of A. calamus promoted wound-healing activity significantly in both the wound models studied. The histological study of the granulation tissue with 20% A. calamus extract ointment-treated animals showed a larger number of inflammatory cells and lesser collagen when compared with the 40% A. calamus extract ointment-treated animals. However, this was better than the control group of animals. Enhanced wound contraction, decreased epithelialisation time, increased hydroxyproline content and histological characteristics suggest that A. calamus extract may have therapeutic benefits in wound healing.

The RP-HPLC method for simultaneous estimation of esomeprazole and naproxen in binary combination

Objective: A simple, precise, reliable, rapid, sensitive and validated RP-HPLC method has been developed to determine esomeprazole magnesium trihydrate (ESO) and naproxen (NAP) in synthetic mixture form. Materials and Methods: Chromatographic separation achieved isocratically on Phenomenex, Luna C18 column (5 μm, 150mm × 4.60mm) and acetonitrile: phosphate buffer (pH 7.0) in the ratio of 50:50 (v/v) as the mobile phase, at a flow rate of 0.5 ml/min. Detection was carried out at 300 nm. The retention times for NAP and ESO was found to be 2.67 ±0.014 and 5.65 ±0.09 min respectively. Parameters such as linearity, precision, accuracy, recovery, specificity and ruggedness are studied as reported in the ICH guidelines. Results: The method was linear in the concentration range of 50-250 μg/ml for NAP and 2-10 μg/ml for ESO with correlation coefficient of 0.999 and 0.998 respectively. The mean recoveries obtained for NAP and ESO were 100.01% and 97.76 % respectively and RSD was less than 2. The correlation coefficients for all components are close to 1. Conclusions: Developed method was found to be accurate, precise, selective and rapid for simultaneous estimation of NAP and ESO.

Development of Spectrophotometric Method for Quantitative Estimationof Amlodipine Besylate, Olmesartan Medoxomil and Hydrochlorthiazide inTablet Dosage Form

A new, simple, accurate, precise and reproducible UV spectrophotometric method is being developed for the simultaneous estimation of amlodipine besylate, olmesartan medoxomil and hydrochlorthiazide in tablet dosage form. The stock solutions were prepared in methanol. The λ max for amlodipine besylate, olmesartan medoxomil and hydrochlorthiazide were 238.5nm, 256.5nm and 271.5nm respectively. The amlodipine besylate, olmesartan medoxomil and hydrochlorthiazide obeyed Beer’s law in concentration range of 5-25μg/ml, 6-30μg/ml and 5-25μg/ ml respectively. Results of analysis of simultaneous equation method were analyzed and validated for various parameters according to ICH guidelines.

Development and validation of reversed phase-high-performance liquid chromatography method for determination of paracetamol and lornoxicam in tablet dosage form.

A simple, precise, reliable, rapid and reproducible reversed phase–high-performance liquid chromatography method was developed and validated for the simultaneous estimation of Paracetamol (PCM) and Lornoxicam (LOX) present in tablet dosage forms. Chromatographic separation achieved isocratically on Luna C18 column (5 μm, 150 × 4.60 mm) and methanol/phosphate buffer (60:40, v/v, pH 7.0) as mobile phase, at a flow rate of 1 ml/min. Detection was carried out at 260 nm. Parameters such as linearity, precision, accuracy, recovery, specificity and ruggedness are studied as reported in the ICH guidelines. The retention times for PCM and LOX was found to be 2.06±0.013and 4.38±0.07 min, respectively. Linearity for PCM and LOX was in the range of 10-50 mg/ml and 8-40 mg/ml, respectively. The mean recoveries obtained for LOX and PCM were 100± 0.16 and 99.50± 0.43%, respectively, and relative standard deviation (RSD) was less than 2. The correlation coefficients for all components are close to 1. The RSDs for three replicate measurements in three concentrations of samples in tablets are always less than 2%. Developed method was found to be accurate, precise, selective and rapid for simultaneous estimation of PCM and LOX in tablets.

Mixed Hydrotropy Solubilization Approach for Quantitative Estimation of Eprosartan Mesylate and Hydrochlorthiazide by UV Spectrophotometer

Two simple, accurate, novel, safe and precise methods were developed for the simultaneous estimation of poorly water-soluble drugs Eprosartan Mesylate and Hydrochlorthiazide in tablet dosage form using 2M Sodium acetate and 8M Urea solution (50:50% W/V) as a mixed hydrotropic solution. Eprosartan Mesylate and Hydrochlorthiazide show maximum absorbances at 267.5 and 271.5 nm respectively. Sodium acetate and Urea solution did not show any absorbance above 240 nm and thus no interference in the estimation of drugs was seen. Eprosartan Mesylate and Hydrochlorthiazide follows the Beer’s law in the concentration range of 15-75 and 5-25 μg/ml (r 2 = 0.9994 and 0.9996). Method-A employs a simultaneous equation method using 267.5 and 271.5 nm as two analytical wavelengths, Method-B, an absorption ratio method, uses 271.5 and 277 nm as two analytical wavelengths for estimation of Eprosartan Mesylate and Hydrochlorthiazide. The optimized methods showed good reproducibility and recovery with ranging from 95.08±0.086 to 99.82±0.097 EPS and HCZ respectively. The developed methods were validated according to ICH guidelines and values of accuracy, precision and other statistical analysis were found to be in good accordance with the prescribed values therefore the both methods can be used for routine monitoring of EPS and HCZ in industry in the assay of bulk drug and tablets.

Eco friendly spectrophotometric method for quantitative estimation of lomefloxacin using hydrotropic approach

The present work describes a novel, accurate, sensitive and economic safe spectrophotometric method was developed by application of hydrotropy, using 8 M Urea solution as hydrotropic solubilizing agent, for the quantitative determination of poorly watersoluble lomefloxacin HCl in tablet dosage form. There were more than 43 times enhancements in the solubility of lomefloxacin HCl increases in hydrotropic solution as compared to solubilities in distilled water. Lomefloxacin HCl shows maximum absorbance at 281 nm. Urea and other tablets excipents did not show any absorbance above 230 nm, and thus no interference in the estimation was seen. Lomefloxacin HCl was obeyed Beer,s law in the concentration range of 5 to 25μg/ml (r2= 0.9998) in hydrotropic solvent with mean recovery ranging from 98.03±0.65 to 98.59±0.32%. Proposed method is new, simple, economic, safe, rapid, accurate and reproducible. The developed methods were validated according to ICH guidelines and values of accuracy, precision and other statistical analysis were found to be in good accordance with the prescribed values. The method can be used for routine analysis in both research laboratories, and pharmaceutical and chemical industries to analyze the drugs without the use of organic solvents thus make the environment eco-friendly.

Bioanalytical Method Development and Validation for the Determination of Levocetirizine in Pharmaceutical Dosage Form and Human Plasma by RP-HPLC -

This study describes the development of a rapid, selective, precise and sensitive reverse phase high-performance liquid chromatography method for the quantitative determination of Levocetirizine Dihydrochloride (LCD) in human plasma and pharmaceutical dosage form. Extraction of drug from plasma was done by employing optimized liquid-liquid extraction procedure. The sample was analyzed using Acetonitrile: Methanol: 20mM Ammonium Acetate Buffer pH-5 (25:55:20 % v/v/v) as mobile phase. Chromatographic separation was achieved on Prontosil C-18 column (4.6 x 250mm, 5μ particle size) as stationary phase using isocratic elution (at a flow rate of 1 mL/min). The peak was detected using UV-PDA detector set at 232 nm and the total time for a chromatographic separation was 8 min. The calibration curve obtained was linear (r2= 0.9998) over the concentration range of 2-10 μg/mL. Method was validated for precision, robustness and recovery. The limit of detection (LOD) and limit of quantitation (LOQ) was 0.0057 and 0.174 µg/mL respectively. There was no significant difference between the amount of drug spiked in plasma and the amount recovered and plasma did not interfere in estimation. Thus, the proposed method is suitable for the analysis of LCD in tablet dosage forms and human plasma.

Simultaneous Estimation of Metoprolol Succinate and Lacidipine in Binary Combination Using High Performance Liquid Chromatographic Method = تقييم في وقت واحد للجمع بين ثنائي السكسينات ميتوبرولول و لاسيدبين باستخدام طريقة الكروماتوجرافي السائلة عالية الأداء

A simple, reliable, rapid, precise, sensitive, rugged and validated RP-HPLC method has been developed to determine Metoprolol succinate and Lacidipine in synthetic mixture form. Chromatographic separation achieved isocratically on Prontosil C18 column (5µm, 250mm × 4.60mm) and acetonitrile: 20mM Phosphate buffer (pH 3.6) in the ratio of 60:40 (v/v) as the mobile phase, at a flow rate of 1 mL/min. Detection was carried out at 278 nm. The mean retention times for metoprolol succinate and lacidipine was found to be 5.25±0.5 and 6.72±0.5 min, respectively. No interference was found by the excipients in the synthetic mixture. Linearity for metoprolol succinate and lacidipine was in the range of 5-25μg/mL and 4-20μg/mL, respectively. The mean recoveries obtained for metoprolol succinate and lacidipine were 100.03 and 99.67 % respectively and RSD was less than 2. The correlation coefficients for all components are close to 1. The proposed method was validated in terms of linearity, range, accuracy, precision, specificity, robustness and the method is successfully applies to the estimation of metoprolol succinate and lacidipine in bulk and in a synthetic mixture.

DEVELOPMENT AND VALIDATION OF A GREEN BIOANALYTICAL METHOD FOR THE DETERMINATION OF SPARFLOXACIN IN PHARMACEUTICAL DOSAGE FORM AND HUMAN PLASMA BY RP - HPLC

This study describes the development of an i nnovative, green , rapid, precise, selective and sensitive reverse phase high - performance liquid chromatography method for the quantitative determination of Sparfloxacin ( SPR ) in human plasma and pharmaceutical dosage form . Extraction of drug from plasma was done by employing optimized liquid - liquid extraction procedure . The sample was analyzed using Methanol : Water ( pH - 7 with triethylamine ) ( 60 : 40 % v / v ) as mobile phase . Chromatographic separation was achieved on Prontosil C - 18 column ( 4 . 6 x 250mm, 5μ particle size ) as stationary phase using isocratic elution ( at a flow rate of 1 ml / min ). The peak was detected using UV - PDA detector set at 254 nm and the total time for a chromatographic separation was 9 min . The calibration curve obtained was linear ( r 2 = 0 . 9998 ) over the concentration range of 5 - 25 μg / ml . Method was validated for precision, robustness and recovery . The limit of detection ( LOD ) and limit of quantitation ( LOQ ) was 0 . 573 and 1 . 542 µg / ml respectively . There was no significant difference between the amount of drug spiked in plasma and the amount recovered and plasma did not interfere in estimation . Other important factors is that the method uses less amounts of organic solvent, produces low levels of was te and does not use buffer solution, minimizing effluent treatment, which contributes to the environment and implements methods aimed green chemi stry, making economic for the industry . Thus, the proposed method is suitable for the analysis of SPR in tablet dosage forms and human plasma .

A Novel Approach using Hydrotropic Solubalization Technique for Quantitative Estimation of Entacapone in Bulk Drug and Dosage Form

PURPOSE Analysis of drug utilized the organic solvent which are costlier, toxic and causing environment pollution. Hydrotropic solution may be a proper choice to preclude the use of organic solvents so that a simple, accurate, novel, safe and precise method has been developed for estimation of poorly water soluble drug Entacapone (Water Solubility-7.97e-(02) g/l). METHODS Solubility of entacapone is increased by using 8M Urea as hydrotropic agent. There was more than 67 fold solubility enhanced in hydrotropic solution as compare with distilled water. The entacapone (ENT) shows the maximum absorbance at 378 nm. At this wavelength hydrotropic agent and other tablet excipients do not shows any significant interference in the spectrophotometric assay. RESULTS The developed method was found to be linear in the range of 4-20 μg/ml with correlation coefficient (r(2)) of 0.9998. The mean percent label claims of tablets of ENT in tablet dosage form estimated by the proposed method were found to be 99.17±0.63. The developed methods were validated according to ICH guidelines and values of accuracy, precision and other statistical analysis were found to be in good accordance with the prescribed values. CONCLUSION As hydrotropic agent used in the proposed method so this method is Ecofriendly and it can be used in routine quantitative analysis of drug in bulk drug and dosage form in industries.