Indo American Journal of Pharmaceutical Research

ISSN NO.: 2231-6876
April 2011

RP-HPLC and chemometric assisted UV-spectrophotometric methods for simultaneous in vitro analysis of atrovastatin calcium, ezetimibe and fenofibrate in their pharmaceutical formulation

Abhishek Pathak1, Sadhana J Rajput*1, Rahul S Gamit2


1. Pharmaceutical Quality Assurance Laboratory,

2. Centre of Relevance and Excellence in Novel Drug Delivery System,

3. Shri G H Patel Pharmacy Building, Pharmacy Department,

4. The Maharaja Sayajirao University of Baroda,


One RP-HPLC and two chemometric assisted UV spectrophotometric methods were developed and validated for the simultaneous in vitro analysis of Atrovastatin calcium (ATV), Ezetimibe (EZET) and Fenofibrate (FEN) in their pharmaceutical preparation and ternary mixtures. The chromatographic separation was achieved on a reversed-phase, Hypersil BDS C8 column (250X4.6 mm i.d, 5  particle size) with a mobile phase consisting of methanol and 0.05M phosphate buffer (pH-6.3 adjusted with sodium hydroxide) in the ratio of (85:15)% v/v. The total run time was 7 min. Quantitation was achieved with UV detection at 248nm based on peak area. Linearity was observed over concentration range of 5 - 12µg mL-1 for ATV and EZET and 80 - 192 µg mL-1 for FEN. The two chemometric methods applied were inverse least square (ILS) and classical least square (CLS). These approaches were successfully applied to quantify each drug in their mixture using the information included in the UV absorption spectra of appropriate solutions in the wavelength range 220-310nm with the intervals of 5nm (Δλ = 5nm) at 19 wavelength points. For the chemometric calibration, 18 ternary solutions were prepared as training set and 10 ternary solutions were prepared as validation set. The developed methods were successfully applied for laboratory prepared mixtures as well as commercial tablet formulation for ATV, EZET and FEN concentration. The results obtained for pharmaceutical formulation by ILS and CLS methods were compared with isocratic HPLC method and a good agreement was found.


Keywords: Atrovastatin Calcium, Chemometric, Ezetimibe, Fenofibrate, Reversed Phase-High Performance Liquid Chromatography (RP-HPLC)




Rajinikar Reddy1*, V.V.Rajesham1, S.Kiran Kumar1, J.Prasanna kumari1, V.Suba2
1 Vijaya College of Pharmacy, Munaganoor, Hayathnagar, Rangareddy(Dist), A.P
2Vel’s College of Pharmacy, pallavaram, Chennai


The present study was designed to investigate anti-arthritic activity of Portulaca olaraceal. sativa leaves in rats. Arthritis was induced in rats in all groups of animals by injecting 0.05 ml of 0.5(w/v) suspension of killed Mycobacterium tuberculosis in paraffin oil into the left hind limb. Preliminary extraction was subjected to acute oral toxicity study according to the OECD guidelines no 423. Based on that, two dose levels i.e. 200 and 300mg/kg were selected for further study. Oral administration of the extract showed the significant results in Freund’s adjuvant induced arthritis in rats. The biochemical study of EEPO (200mg/kg & 300mg/kg) showed the increase WBC count was significantly suppressed and standard diclofenac sodium (P<0.01).The increased lymphocyte count in adjuvant control group was significantly restored back to normal by test and standard drug (P<0.01).There is a decreased protein content in standard and extract drug treatment group as compared to adjuvant control group.


Simultaneous Resolution of Clopidogrel Bisulphate and Aspirin in their Combined Dosage Form by Ratio First Derivative Spectrophotometry

Ruikar Dipti B*, Sadhana J Rajput

Pharmaceutical Quality Assurance Laboratory, Pharmacy Department, Faculty of Technology and Engineering, The M. S. University of Baroda, Vadodara - 390001, India


In the present work, the ratio spectra first derivative spectrophotometry is proposed for the resolution of Clopidogrel bisulphate (CLP) and Apirin (ASP) in synthetic mixtures prepared in random fashion and tablets available in 1:1 and 1:2 ratio. The calibration graph follows Beer’s law in the range of 6 to 30 μcg/ml for clopidogrel bisulphate and from 4-20 μcg/ml for aspirin. The mean recoveries & relative standard deviations were found as 99.40% & 0.442% for CLP and 100.09% and 0.905% for ASP, respectively in 1:1 ratio. Similarly the mean recoveries & relative standard deviations were found as 999.84% & 1.024% for CLP and 99.95% and 0.578% for ASP, respectively in 1:2 ratio. The graphical treatment of the overlapping spectra depends on division of the absorption spectrum of the binary mixture by a standard spectrum of one of the components and then calculating the first derivative of the ratio spectrum. These approaches were successfully applied to quantify CLP combined with ASP in tablets respectively and validated accordingly for random ratios in synthetic mixtures.


Anthelmintic activity of root of Benincasa hispida (Petha)

Chiranjib Bhattacharjee*, Subal Debnath, G. Ganesh Kumar, Santhosh Kumar C, T. Shivaraj Gouda
Srikrupa Institute of Pharmaceutical Sciences, Vil. Velkatta, Kondapak (mdl), Dist. Medak, Siddipet. Andhra Pradesh – 502 277, India


The present study was undertaken to evaluate anthelmintic activity of crude aqueous, Petroleum ether, chloroform and methanol extracts of Benincasa hispida (Petha) root using Pheretima posthuma as test worms. Single concentration (5%) of extracts were tested in the bioassay, which involved determination of time of paralysis (P) and time of death (D) of the worms. Piperazine citrate was included as standard reference and distilled water as control. The results of present study indicated that Benincasa hispida (Thumb) root extracts were exhibited anthelmintic activity significantly (***p<0.001) when compared with standard (Piperazine citrate) group. Further studies are in process to isolate the active principles responsible for the activity.



Santosh Kumar .C1*, Chiranjib Bhattacharjee1, Subal Debnath1, Atul N. Chandu
1Srikrupa Institute of Pharmaceutical Sciences, Vil. Velkatta, Kondapak (mdl), Dist. Medak, Siddipet,
Andhra Pradesh – 502 277
2 University institute of pharmacy, Pt. Ravishankar Sukla University, Raipur, C.G., India


The objective of the present study was to evaluate the synergistic antimicrobial and anti-tubercular activities of phyllanthus acidus Methanolic extract. Antibacterial study was carried out by plate hole diffusion or agar well diffusion assay to determine the growth inhibition of bacteria. Antifungal activity was performed by the use of Saubouraud dextrose agar medium (SDA). The synergistic activity study was calculated by means of cup plate method (Kirbauy bauer technique) using two wells in a plate. The Methanolic plant extract of Phyllanthus acidus (500μg/ml) was used in combination with
Oxytetracycline (500μg/ml). Anti-tubercular assay was performed using Micro plate Alamar Blue Assay (MABA) using the suspension of Mycobacterium tuberculosis H37Rv strain. The concentrations of plant extract were used are 1000 μg/ml, 500μg/ml, 250μg/ml, 125 μg/ml, 62.5μg/ml. The medicinal plant appear to have a broad antimicrobial activity spectrum, they could be useful in antiseptic and disinfectant formulation as well as in anti-tubercular activity. Among the various micro organisms, the Methanolic extract was more active against Micrococcus flavum. In antifungal activity the Methanolic
extract shows positive results for all fungus. The anti-tubercular activity was compared with standard drug Rifampicin. The Methanolic extract was having more percentage inhibition when compared to other extracts.


Simultaneous Estimation of Abacavir Sulfate and Lamivudine in their Combined Dosage Form by First Derivative Zero-Crossing and Ratio First Derivative Spectroscopic Methods

Quality Assurance Laboratory, Centre of Relevance and Excellence in Novel Drug Delivery System,
Pharmacy Department, G. H. Patel Building, Donor’s Plaza,The Maharaja Sayajirao University of Baroda, Fatehgunj,Vadodara, Gujarat, India – 390 002


Two simple, sensitive, specific and accurate spectrophotometric methods are proposed in the present work for the simultaneous estimation of Abacavir sulfate (ABA) and Lamivudine (LMV) in their binary mixture. In first derivative zero crossing technique the absorbances at 234.2 nm (zero crossing point of LMV) were plotted against the respective concentrations of ABA. Similarly the absorbances at 287.3 nm (zero crossing point of ABA) were plotted
against the respective concentrations of LMV. For the ratio spectra first derivative spectrophotometric method, the graphical treatment of the overlapping spectra depends on division of the absorption spectrum of the binary mixture by a standard spectrum of one of the components and then the first derivative amplitudes measured at 233.0nm for ABA and 236.4nm for LMV .The calibration curves follow Beer’s law in the range of 5 to 30 μg/ml for ABA and 2.5 to
15 μg/ml LMV for both the methods. The methods were successfully applied to quantify ABA combined with LMV in tablets and validated accordingly.



Nilesh P Babre1*, Santhosh Kumar.C1, Subal Debnath, M N. Arjun1, Shravan Reddy2
1Srikrupa Institute of Pharmaceutical Sciences, Vil. Velkatta, Kondapak, Dist. Medak, Siddipet. Andra Pradesh. – 502 277.
2M.S.Ramaiah College of Pharmacy, Bangalore-560054


Many hepatoprotective herbal preparations have been recommended in alternative system of medicine for the treatment of hepatic disorders. No
systemic study has been done on protective efficacy of Crossandra Undulaefolia belonging to family Acanthanceae to treat diseases. The Hydro-Ethanolic extracts of aerial parts of Crossandra Undulaefolias (ECU). The animals were weighed each and divided in group of five. Liver damage was achieved by injecting Paracetamol (2 g/kg). The treatment groups pretreated with ECU Group-IV (200mg/kg) and Group- V (400mg/kg). Silymar in was used as reference standard drug. At the end of 7 days, blood was collected, liver extracted, weighed, processed for histopathological assessments and alteration in the levels of biochemical markers of hepatic damage like serum transaminases (AST,ALT), alkaline phosphatase (ALP) , bilirubin  cholesterol, high density lipoprotein (HDL) and tissue GSH were tested in all the groups. The ECU exhibited a significant hepatoprotective effect by lowering the elevated serum levels of AST, ALT, ALP, total and direct serum bilirubin, cholesterol and significantly (p<0.05) increased HDL and moderately increased total protein and albumin in a dose dependent manner These biochemical observations were supplemented by treatment of liver toxicity in traditional medicine. Histopathological examination of liver sections. These findings suggest the use of this plant for itsHepatoprotective effects.


Formulation and Evaluation of Oro Dispersible Tablets of Famotidine using Superdisintegrants

*Vyomesh N. Raval, Gajanan J. Deshmukh, A. K. Seth. Tejas K Ghelani, Sharad Kumar, Hemangi Patel, Sachin Chauhan
Department of Pharmacy, Sumandeep Vidyapeeth, Vadodara, Gujarat, India.


The objective of the present investigation was to prepare oro dispersible tablets of Famotidine, because of their convenience in administration and suitability for patients having dysphagia. Tablets were prepared by direct compression using sodium starch glycolate and croscarmellose sodium and crospovidone as superdisintegrants. Microcrystalline cellulose pH-102 was used as diluent and mannitol, sodium saccharine to enhance the organoleptic properties of tablets. The tablets were evaluated for weight variation, hardness, friability, assay, in-vitro disintegration time, in-vitro dissolution study. Hardness and friability data indicated good mechanical strength of tablets. The results of in-vitro disintegration time indicated that the tablets dispersed rapidly in mouth within 60 seconds. It was concluded that superdisintegrants addition technique is a useful method for preparing oro dispersible tablets by direct compression method.


Polymeric Micelles: A Nanoscience Technology

B. Raja Rajeshwar1*, Anvesh Gatla1, Gaddam Rajesh2*, N. Arjun2, M. Swapna2
1 Department of pharmaceutics, CMR College of pharmacy, Hyderabad
2 Department of pharmacology, Srikrupa institute of pharmaceutical sciences, Siddipet, Medak, Andhra
Pradesh, India -502277.


One of the most widely studied subjects in nanoscience technology is related
to the creation of supramolecular architectures with well-defined structures
and functionalities. In the late 1960s, micelles drew much attention as drug
carriers owing to their easily controlled properties and good
pharmacological characteristics. These supramolecular structures are
generated as a result of self-assemblage of amphiphilic block polymers.
Self-assembly of block polymers via hydrophobic and hydrophilic effects,
electrostatic interactions, hydrogen bonding, and metal complexation has
shown tremendous potential for creating such supramolecular structures
with a wide array of applications. Polymeric micelles have gathered
considerable attention in the field of drug and gene delivery due to their
excellent biocompatibility, low toxicity, enhanced blood circulation time,
and ability to solubilize a large number of drugs in their micellar core. In
this article we have reviewed several aspects of polymeric micelles
concerning their general properties, preparation and characterization
techniques, and their applications in the areas of drug and gene delivery.
Polymeric micelles can be used as 'smart drug carriers' for targeting certain
areas of the body by making them stimuli-sensitive or by attachment of a
specific ligand molecule onto their surface.



Dhanyakumar  Chougule*, Nilofar  Naikwade 

Appasaheb Birnale College of Pharmacy, Sangli


Ribavirin is an antiviral agent commonly used for the treatment of severe acute respiratory syndrome. The present study investigated the stability of marketed oral Ribavirin syrup. The syrup was subjected to accelerated stability testing at 40°C and 75% RH for 6 months as per ICH guidelines. HPLC method to estimate Ribavirin was developed and validated. Employing a validated high-performance liquid chromatographic method, the Ribavirin content of the syrup has been demonstrated to exhibit changes throughout the storage period. No physical changes was observed in syrup suggesting that it remained physically stable, but effected the chemically stability of syrup under the stated conditions.