ABSTRACT A simple, precise, rapid, specific and accurate reverse phase high performance liquid chromatography method was developed for simultaneous estimation of Paroxetine and Clonazepam in pharmaceutical dosage form. Chromatographic separation was performed on Agilent Eclipse XDB (C8) (4.6mm x 150mm, 5m) column, with mobile phase comprising of mixture of buffer (pH7, adjusted with ammonium acetate), acetonitrile in the ratio of 82:18v/v, at the flow rate 0.8 ml/min. The detection was carried out at 265 nm. The retention times of paroxetine and clonazepam were found to be 2.36 and 3.14 mins respectively with a run time of 5 mins, theoretical levels for paroxetine and clonazepam were 6753 and 4693 respectively, with a resolution of 5.10. As per ICH guidelines the method was validated for linearity, accuracy, precision, limit of detection and limit of quantitation, robustness and ruggedness. Linearity of paroxetine was found in the range of 100-300μg/mL and that for clonazepam was found to be 2-6 μg/mL. Keywords: Paroxetine, Clonazepam, RP-HPLC, Method development, Validation.

ABSTRACT The goal of the present investigation was carried out to develop a validated analytical RP-HPLC method for simultaneous separation and estimation of Lamivudine and Zidovudine for pharmaceutical formulations. The chromatographic separation was accomplished on Welchrom RP-C18 Column (250 mm X 4.6 mm; 5μm), Shimadzu LC-20AT Prominence Liquid Chromatograph and with a mixture of 10 mM Phosphate buffer (pH 3.6): acetonitrile (50:50, v/v). The flow rate was fixed at 1.2 mL/minute and the analysis was performed using Shimadzu SPD-20A Prominence UV-Visible detector at 241 nm. The Lamivudine and Zidovudine were separated within three minutes. The retention time for Lamivudine and Zidovudine was found to be 2.393 minutes and 2.7 minutes respectively. The calibration plots were linear over the concentration range of 2-10 μg/mL for Lamivudine (r2=0.9995) and 4-20 μg/mL for Zidovudine (r2=0.9991). There was no interference due to commonly used excipients. The % RSD for both the drugs were calculated and showed less than 2% which obviously indicates that the present method was said to be highly precise. Regarding accuracy of the developed method the %RSD were also found less than 2% which shows the method is completely accurate. The method was very sensitive with regard to limit of detection and limit of quantitation for Lamivudine and Zidovudine were found to be 0.181 μg/mL, 0.576 μg/mL and 0.551 μg/mL, 1.747μg/mL respectively. The mean assay values for Lamivudine and Zidovudine were arrived at 98.88% and 99.70 % respectively. The developed RP HPLC method was found to be simple, rapid, sensitive, highly precise and accurate and can be successfully employed for simultaneous estimation of Lamivudine and Zidovudine in pharmaceutical formulations. Keywords: RP HPLC Method, Simultaneous estimation, Lamivudine, Zidovudine.

ABSTRACT A simple, precise, rapid, and accurate reverse phase high performance liquid chromatography method was developed for the estimation of Bromfenac in Rabbit plasma by using Paracetamol as IS. The protein precipitation technique was used for the extraction of analyte from rabbit plasma. The chromatographic separation was performed on Agilent, Zorbas SB C18 (4.6mm×250mm×5μ) column with PDA detection at 267nm. The mobile phase comprising of mixture of methanol and buffer 0.1% ortho phosphoric acid (pH 2.6) in the ratio of 30:70v/v, at the flow rate 0.8ml/min. The analytical method described here is valid for the estimation of Bromfenac in Rabbit plasma over a range of 50μg/ml-150μg/ml. The retention times of Bromfenac and Paracetamol were found to be 3.781 and 2.470mins respectively. The correlation coefficient for Paracetamol and Bromfenac were 0.998 and 0.999 respectively. The LOD and LOQ values for Bromfenac were found to be 0.13μg/ml and 0.40μg/ml respectively. %RSD of Bromfenac and Paracetamol was obtained to be 0.41and 0.28 respectively. This method is time efficient and samples are easy to monitoring Bromfenac in plasma. Keywords: HPLC-PDA method, Rabbit Plasma, Bromfenac, Protein precipitation.

ABSTRACT

Analytical methods development must be validated to provide reliable data for regulatory submissions. Method development is the process of proving that an analytical method is acceptable for use to measure the concentration of an API in a specific compounded dosage form which allow simplified procedures to be employed to verify that an analysis procedure, accurately and consistently will deliver a reliable measurement of an active ingredient in a compounded preparation.These methods are essential for a number of purposes, including testing for quality control release, testing of stability sample, testing of reference materials and to degree of assurance and is an important process in the drug discovery. Although the drug shows good potency, lack of validated analytical method will not allow the drug to enter into the market. This is to ensure the quality and safety of he drug. This review gives ideas about various methods to check the stability of dug and various validation parameters as per various regulatory authorities. Keywords: Method Development, Drug analysis, Validation, ICH.

ABSTRACT A simple, precise, rapid, specific and accurate reverse phase high performance liquid chromatography method was developed for simultaneous estimation of Trifluoperzine and Isopropamide in pharmaceutical dosage form. Chromatographic separation was performed on Agilent zorbax SB-C18, 4.6 x 250mm, 5micros column, with mobile phase comprising of mixture of buffer (pH 6.0, adjusted with Ortho phosphoric acid, Acetonitrile in the ratio of 80:20v/v), at the flow rate 0.8ml/min. The detection was carried out at 227nm. The retention times of Trifluoperzine and Isopropamide were found to be 2.4 and 3.6mins respectively with a run time of 10mins, theoretical levels for Trifluoperzine and Isopropamide were 5194 and 6738 respectively, with a resolution of 7.6. As per ICH guidelines the method was validated for linearity, accuracy, precision, limit of detection and limit of quantitation, robustness and ruggedness. Linearity of Trifluoperzine and Isopropamide was found in the range of 30-130μg/ml and that for Isopropamide was found to be 150-250μg/ml. The correlation coefficient for Trifluoperzine and Isopropamide were 0.999 and 1 respectively. The LOD values for Trifluoperzine and Isopropamide were 2.963 and 2.9851 respectively. The LOQ values for Trifluoperzine and Isopropamide were and 9.877μg/ml and 9.9502μg/ml respectively. This demonstrates that the developed method is simple, precise, rapid, selective, accurate and reproducible for simultaneous estimation of Trifluoperzine and Isopropamide tablet dosage form.

Keywords: HPLC method, Stelbid, Validation, analytical method optimization.

ABSTRACT Analytical methods are developing rapidly, and it is considered to be a small-scale activity, but this is not always true in the case of controlling and monitoring laboratories whose number of runs performed is high. This makes an analytical laboratory comparable with the fine chemicals or pharmaceutical industry. Hence new analytical methodologies are being developed, according to green chemistry standards and these chemical processes are evaluated for their effects on the environment. Furthermore, most of these techniques can be automated and quite easily coupled with ―green‖ methods. Introducing these techniques to everyday practice intolaboratories is an important step in diminishing the negativeeffects of analytical chemistry on the environment. Keywords: Green Chemistry, Supercritical Fluid Extraction, Solid Phase Extraction.

ABSTRACT Fenofibrate which is chemically propan-2-yl 2-{4-[(4-chlorophenyl) carbonyl] phenoxy}-methyl propanoate. It is mainly used to reduce cholesterol levels in patients at risk of cardiovascular disease. Like other fibrates, it reduces low-density lipoprotein (LDL) and very low density lipoprotein (VLDL) levels, as well as increasing high density lipoprotein (HDL) levels and reducing triglycerides levels. It also appears to have a beneficial effect on the insulin resistance featured by the metabolic syndrome. . Literature survey reveals that Fenofibrate is estimated individually by UV spectrometry, RP-HPLC, LC/MS, LC/MS/MS and stability indicating HPLC. Keywords: Fenofibrate, UV spectrometry, RP-HPLC, LC/MS, LC/MS/MS.

ABSTRACT Atomic emission spectroscopy (AES) is a method of chemical analysis that uses the intensity of light emitted from a flame, plasma, arc, or spark at a particular wavelength to determine the quantity of an element in a sample. The wavelength of the atomic spectral line gives the identity of the element while the intensity of the emitted light is proportional to the number of atoms of the element. Atomic emission spectroscopy that concerns the emission of radiation by the suitably excited atomic vapors of the analyte. Here, the emitted radiation and its intensity form the basis for the qualitative and quantitative applications of the technique. It is a multi-element analytical technique that can be used for the analysis of materials in gaseous, liquid, powder or solid form. Its high detection power and wide variety of excitation sources makes it the most extensively used method for analysis. Of the various different atomic emission spectroscopic, an inductively couple plasma (ICP) acts as an atomization- excitation source; the technique being called ICP-availability of sufficiently sensitive interference free spectral lines. Keywords: Inductively coupled plasma (ICP), Atomic emission spectroscopy (AES), X-Ray flouroscence (XRF), Radio frequency (RF), Charge coupled devices (CCDs) or Charge injection devices (CIDs).

ABSTRACT

Antioxidant enzymes are known defense molecules, which eliminate reactive oxygen species (ROS) and free radicals that damage useful enzymes in the cell under various stress conditions. Significant efforts have been made to explore these antioxidant enzymes as a potential candidate for the diagnosis of certain diseases. Hypobaric hypoxia is a condition of oxidative stress leading to various diseases through the involvement of free radicals. In the present study, we have analyzed the expression of antioxidant enzymes in the plasma of male albino rats (185 ± 10 g) that were segregated into hypobaric-hypoxia susceptible (HHS) and tolerant (HHT) on the basis of their ‘gasping time’. After one week of normalization, both HHS and HHT rats were exposed to 30,000 ft for 1 h and sacrificed to collect the blood sample. Catalase (CAT) and peroxidase showed 3.9 and 2 fold increased enzymatic activity, respectively in HHT rats as compared to HHS rats. There was no significant difference in superoxide dismutase (SOD) activity in HHT when compared with HHS rats. Differential activity of CAT and peroxidase in HHS and HHT rats under hypoxia stress was also confirmed by zymography. Thus differential activity of these antioxidant enzymes in HHS and HHT may have a regulatory role under hypobaric hypoxic conditions.

Keywords: Antioxidant enzymes, Catalase, Peroxidase, Hypobaric hypoxia, SOD.