International Journal of Pharmaceutical Sciences and Drug Research

308-314

Authors: D. Nagasamy Venkatesh*, H. C Kiran, S. Shashikumar, V. Indurekha, K. J Thirumalai Subramaniam, A. Uma Rani

Dendrimers are synthetic, highly branched, monodisperse macromolecules of nanometer dimensions with exact and large number of functional groups, distributed with unprecedented control, makes them a promising scaffolds, for drug delivery. Dendrimers serves as an ideal vehicle for cancer therapy, immunology, vaccines, antivirals, biosensors for diagnostics, neuron capture therapy, photodynamic therapy and photo thermal therapy. Dendrimers chemistry is one of the most fascinating and rapidly expanding areas in the field of chemistry. Prior to the dendrimer technology, nanoparticle drug delivery systems were one of the choicest systems owing to their selectivity and stability of therapeutic agents incorporated into the system. However, few drawbacks such as reticuloendothelial system (RES) uptake, drug leakage, immunogenicity, hemolytic toxicity, cytotoxicity, hydrophobicity etc., impede the usage of these nanostructures. Further, these shortcomings shall be circumvented by modifying the surface engineering, such as poly ester dendrimer, arginine dendrimer, glycol dendrimer, PEGylated dendrimers etc., Unique properties of uniform size, water solubility, modifiable surface functionality and availability of internal cavities makes them intriguing carrier for biological and drug delivery system. In the present review, we focused on the bioactive agents that can be easily encapsulated into the interior cavity (or) chemical attachment, conjugation (or) physically adsorbed on to the dendrimer surface to serve the desired properties of the carrier to cater specific needs of the active components, its characterization and application.

321-328

Authors: Karva G. S.1*, Katariya V. R.2, Shahi S. R.1

The aim of present work was to improve the dissolution rate of Ritonavir by preparing micro-particles with certain hydrophilic polymers via spray drying technique. Role of spray drying method was studied for solubility enhancement of poorly aqueous soluble model Ritonavir using solid dispersion approach. All the carriers showed dissolution improvement vis-à-vis pure drug in varying degrees. The solid dispersions were characterized in comparison with pure drug and corresponding Physical mixture (PM). The prepared solid dispersion were evaluated by drug content analysis, saturation solubility, DSC (Differential scanning calorimeter), SEM (Scanning electron microscopy), PXRD (Powder X-ray diffraction), FTIR (Fourier transform infrared spectroscopy).and in vitro drug release. Absence of pure drug peaks in PXRD suggests transformation of crystalline drug into an amorphous form. The transformation of Ritonavir from crystalline to amorphous state by spray drying and the hydrophilic coating of drug particles by the polymers are considered for improvement of Solubility and dissolution of Ritonavir.

334-339

Authors: Dharmik D Patel1*, Ketan P Modi2, Anil K Patel3

The present study was conducted to find out the changes in immunosuppressant drug prescription pattern and trends in kidney transplant patients. In this study 613 Indian transplant patients who underwent kidney transplantation between July 2004 and June 2011 were enrolled. Various data of all transplant patients including immunosuppressant drug medication, changes in the prescription, use of antibody for induction and antirejection treatment were collected during their hospital stay and ambulatory visit. Antibody use as an induction agent has increased from during the study period. Induction was used in 23.1% during the year 2005 and increased to 44.4% during the year 2009. Among induction agent ATG was most commonly preferred agent, followed by daclizumab and basiliximab (24% ATG Vs 6.9% daclizumab Vs 6.9% basiliximab). Use of tacrolimus has increased (94% on tacrolimus vs 6% on cyclosporine in 2010). Mycophenolate mofetil is most commonly used antiproliferative agent (80% on MMF vs 20% on azathioprine in 2010). Trend is towards more use of MMF though azathioprine is being used in significant number of patients. Analysis of maintenance immunosuppression after renal transplant showed 60% patient maintained their original regimen over 7 years of follow up. Sirolimus was introduced in 1.5 to 7% patients during follow up period. Anti rejection treatment was required in 22-47% renal transplant recipients and trend towards decreasing rejection episode was seen. Steroids were used in the treatment of rejection in 90-100% patients. Use of ATG for treatment of rejection has increased from 11.5% in 2005 to 40% in 2010-11. By this study we conclude that immunosuppressant drugs have passed through significant changes during the year 2005 to 2011. Use of ATG as an induction agent and for treatment of rejection has increased. Similarly MMF and tacrolimus are most commonly used in maintenance regimen for renal transplant patients.

345-348

Authors: Praveen R.P.*, Ashalatha S Nair

Myxopyrum smilacifolium is a large woody climbing shrub belonging to the family Oleaceae which is known for many medicinal properties. In the present study, initially a short term cytotoxic study (3 hours) with hepatocarcinoma cell line HepG2 using Trypan Blue Assay was carried out against methanolic extracts of root and callus and found an appreciable result for both extracts. Then the study was extended to the same cell line employing MTT assay. In this procedure extracts were subjected to 24 hours treatment and cytotoxicity was measured using a colorimetric method based on the ability of metabolic active cells to cleave the yellow tetrazolium salt MTT to an insoluble purple formazan crystal. A concentration dependent activity was observed in both Trypan Blue as well as MTT assay. The results showed that both the extracts possess potent cytotoxic activity. IC50 value from MTT assay was found to be 63.75µg/l and 98.75µg/l for root and callus extracts respectively. To the best of our knowledge, this is the first study to demonstrate the cytotoxic potential of callus extracts of Myxopyrum smilacifolium Blume.

354-360

Authors: Samir Baniya1*, Dhananjaya D.R.1, Ankit Acharya2, Bishal Dangi1, Arjun Sapkota1

The present study was designed to evaluate the cardioprotective potential of Citrus grandis (L.) Osbeck (CGO) peel extract in rats. Citrus grandis (L.) Osbeck peel extract was evaluated for protection against cyclophosphamide (200 mg/kg body wt., i.p.) and doxorubicin (15 mg/kg body wt., i.p.) induced cardiotoxicity in male albino rats. Biomarkers like lactate dehydrogenase (LDH), alanine aminotransaminase (ALT), aspartate aminotransaminase (AST), alkaline phosphatase (ALP), total cholesterol (TC), triglyceride (TG) and creatinine kinase (CK-MB) along with heart weight index and antioxidant enzymes was considered to determine the cardioprotective property. Histopathological study was also carried out on heart of experimental animals. The CGO peel extract was found to contain alkaloids, flavonoids, steroids and triterpenoids, saponins, phenolic compounds and tannin as chemical constituents. Cyclophosphamide (CYP) and doxorubicin (DOX) treated groups exhibited significant increase in LDH, ALT, AST, ALP, TC, TG and CK-MB level and decrease in catalase (CAT), superoxide dimutase (SOD) when compared to control group. Pretreatment with different doses of CGO significantly reduced the serum biomarkers and increased the tissue antioxidant level when compared to DOX and CYP alone treated groups. Moreover, treatment with CGO also improved CYP induced changes in histopathology of heart which may be due to its antioxidant property. The Citrus grandis (L.) Osbeck exerted protective effect against CYP and DOX induced cardiotoxicity in rats, which may be due its lipid lowering and antioxidant properties. These findings might be helpful to understand the beneficial effects of CGO extract against myocardial injury although further study is needed to confirm its mechanism.

365-369

Authors: Parth U. Patel*, T. Y. Pasa, Khushbu A. Thakor, Ruchita J. Chauhan

A stability indicating RP-UPLC method was developed and validated for simultaneous estimation Ofloxacin and Ornidazole in tablet dosage form. The separation was achieved under optimized chromatographic condition on a Waters® C18 Acquity UPLC BEH (100 mm × 2.1 mm, 1.7μm) column with mobile phase consist of Water: Acetonitrile: Triethylamine in the ratio of 85: 15: 0.1 v/v. An isocratic elution at a flow rate of 0.2 ml/min at ambient oven temperature was carried out with PDA detection at 300 nm. The retention time for Ofloxacin and Ornidazole was 3.9 min and 6.4 min respectively. The degradation was observed under acidic, alkali, oxidative, photolytic and thermal conditions. The linearity was found to be in the concentration range of 50-150μg/ml for Ofloxacin and 125-375μg/ml for Ornidazole. The % recoveries at 50% were found to be 100.46% & 100.22% for Ofloxacin & Ornidazole respectively. The % recoveries at 100% were found to be 99.70% & 99.83% for Ofloxacin & Ornidazole respectively. The % recoveries at 150% were found to be 99.67% & 100.30% for Ofloxacin & Ornidazole respectively. The method was validated as per ICH guideline and the values were found to be within the limits. So, the proposed method was found to be simple, linear, accurate, precise, stability indicating, robust and specific.

211-228

Authors: C. S. Barik1*, S. K. Kanungo1, N. K. Tripathy2, J. R. Panda3, M. Padhi4

Nature has been a source of medicinal agents for more than thousands of years and herbal therapy predominates in traditional systems of medicine as well as in alternative medicine practiced in various cultures such as Indian system of medicine, Chinese Traditional Medicine, Unani classical literature. The present review deals with various polyherbal formulations used by different countries of the world. Information on traditional herbal formulations was documented in the form of research and review articles in various journals. The aim of this review is to summarize the different types of herbs used for the preparation of polyherbal formulations, their therapeutic potentials including clinical and preclinical results along with their safety and efficacy. This review will facilitate to gain all about the past scientific research and the necessary information about the enormous pharmacological activities of polyherbal formulations which will insist the young researchers for future research to protect human beings from various types of diseases and may serves as a natural gold for the promotion of mankind.

235-243

Authors: Ganesh. M1, Ramesh. K2*, Madhusudan Rao. Y3, Chandra Shekar. B4

In present study, immediate release solid dispersion formulation of Entacapone with various lipids was developed using spray drying method. Entacapone is indicated for the treatment of Parkinson’s disease (PD) as an adjunct to levodopa/carbidopa therapy. Based on the process feasibility and solubility of resulting spray dried powder, formulation ENSD10 was selected for characterization and analyzed for in vitro dissolution profiles in three different pH media. The particle size of Entacapone in spray dried formulation ENSD10 was drastically reduced down to d90 of 25.86µ against d90 of as such drug substance, i.e., 124.74µ. Analysis by differential scanning calorimetry showed that the percentage of crystallinity has been significantly reduced in formulation ENSD10.The optimized solid dispersion formulation (ENSD10) by spray drying method comprising drug to lipid to lipid carrier ratio of 0.5:1:3 (Entacapone: Gelucire 50/13: Compritol) enhanced solubility nearly 6.31 fold and 3.75 fold as compared to pure drug and spray drug alone, respectively. The rate of drug release from spray dried powder was significantly improved and found to be on higher side as compared to the drug release from as such drug and spray dried drug alone. The obtained results suggested that developed Entacapone along with lipid carrier mixture by spray drying method has promising potential for oral delivery and might be an efficacious approach for enhancing the therapeutic potential of active substance.

251-258

Authors: Sija. S. L1*, Potty. V. P2, Santhoshlal. P. S1

Anacardium occidentale L. is much known for its commercially valuable kernals and also bioactive compounds like polyphenols which serve as natural antioxidant having an ability to quench cytotoxic free radicals that may prevent many diseases. The present study was carried out to evaluate the presence of anacardic acid, total phenolic compound, antioxidant activity and antibacterial activity in methanolic extract of callus and various plant parts such as flower, young leaves, shoot and cotyledon. Total phenolic contents were measured by the Folin-Ciocalteau method using gallic acid as standard compound. The maximum phenolic content was found in the methanol extract of flower (260.8 ± 5.543 mg/g) whereas the methanol extract of cotyledon (16.2 ± 3.284 mg/g) showed the lowest total phenolic content. The action of 1, 1-diphenyl-2-picrylhydrazyl (DPPH) on radical scavenging effect of the extracts was determined spectrophotometrically using ascorbic acid as standard compound. All extracts exhibited a DPPH radical scavenging activity and among the extracts, A. occidentale flower demonstrated greater antioxidant potential with a low IC50 (206.312 ± 0.38µg/ml) in comparison with those of the other extracts. Total phenolic contents showed positive correlations with the DPPH radical scavenging activity (r = 0.979; p<0.01) and negative correlations with IC50 (r = -0.984; p<0.01). The antibacterial capabilities of various extracts were also investigated against two common human pathogens of clinical importance, Escherichia coli and Staphylococcus aureus. Presence of anacardic acid in various extracts was also estimated by using HPTLC method. Pre-coated aluminum silica gel plate 60F254 was used as stationary phase and chloroform: ethyl acetate (9:1) was used as mobile phase.

263-269

Authors: Sayanti Sau*, Mukund Handral

Dalbergia sissoo (family: Fabaceae) is an Asian deciduous rosewood tree. It is the state tree of Punjab state (India) called as Shisham used for antipyretic, emesis, ulcers, leucoderma, stomach troubles and skin disease, memory enhancer etc. The aim of this study is to evaluate the learning and memory activity of ethanolic leaf extracts of Dalbergia sissoo in mice. The animals were divided into six groups of 6 each. Group I was considered as normal control, II as Standard control (Imipramine), III, IV and V was treated group (ELDS 300, 450 and 600 mg/kg respectively) and VI as scopolamine treated (negative control).The effect of ethanolic leaf extract of Dalbergia sissoo was investigated in mice for memory enhancing activity using various experimental paradigms of learning and memory viz. Transfer latency (TL) on elevated plus maze and passive avoidance. For memory and learning activity vehicle/ extracts / STD drug administered daily for first seven days, on 8th day dementia was induced by scopolamine. ELDS significantly enhanced the learning and memory activities against the scopolamine induced dementia and significant decrease in Acetylcholinesterase level in brain in animals. The memory enhanced activity as evidenced by learning and retrieval was due to cholinergic facilitatory effect in animals. These results indicate a possible memory enhancing action of Dalbergia sissoo which qualitatively comparable with that of piracetam.

275-278

Authors: Mohd Aslam1, Mohd Tariq2*, Katheem M. Farhan3, Mohd Aftab Ahmad2, Reshma Jolly4, Aisha Siddiqui1

Kushta Marjan (KM) is one well-known formulation prepared from coral used by Unani physicians for effective treatment of a various ailments. Coral is the calcareous skeleton of the minute marine organism and belongs of phylum coelenterate and is used internally in the form of kushta. It is highly efficacious in Cough, Asthma and Anorexia. Till date no scientific work has been performed to establish quality control of KM. In this study we have enlightened the preparation procedures, structural, functional and physico-chemical properties of KM by analyzing it on classical tests along with modern scientific techniques like FTIR (Fourier-transform infrared spectroscopy), TGA (Thermogravimetric Analysis) and DSC (Differential Scanning Calorimetry) to establish its quality control parameters. FTIR spectra showed peaks attributed to calcium oxide, organic matter and several other substances. Total weight loss during TGA was 30.09% and DSC curve showed peaks at 20°C, 65°C, 183°C and 261°C. Since the study has been first of its kind the results obtained might be taken as standard finger print of kushta marjan and might be helpful for further formulation and standardization of KM in routine analysis.

284-289

Authors: S. V. Kulkarni, M. C. Damle*

Embelin, a naturally occurring benzoquinone, is obtained from fruits of Embelia tsjeriam cottam, which plays a vital role in the Ayurvedic system of medicine. An approach for the stress degradation was successfully applied for the development of stability indicating HPTLC method for the determination of Embelin. It was spotted on the plates precoated with silica gel 60 F254 and developed using toluene: ethyl acetate: formic acid, (6:3.5:0.5 v/v/v) as mobile phase. Densitometry analysis was carried out at 291 nm. The method showed high sensitivity with good linearity over the concentration range of 200-1000ng/spot with correlation coefficient value 0.997. The accuracy of the method was established based on the recovery studies. The LOD and LOQ were 6.09ng/band and 18.48ng/band respectively. The system showed a peak for Embelin at Rf of 0.68 ± 0.02.The aim of our study was to observe the effect of various stress conditions on this potential drug candidate. Stress testing of Embelin was carried out according to the International Conference of Harmonization (ICH) guideline Q1A (R2). Embelin was subjected to stress conditions of acid, alkaline hydrolysis, oxidation, photolysis and thermal degradation. A significant degradation was found to occur by acid and oxidation hydrolysis and to a lesser extent under thermal stress and alkaline hydrolysis; and much lesser with photolytic stress. Stress degradation studies on embelin provide an insight into its stability.

298-303

Authors: S. Manivannan1*, P. Madhavi1, S. Bhuvaneswari2

Alpha amylase is one of the important and well known industrial enzymes that cause the breakdown of starch or glycogen. Use of microorganism for the production of amylase is economical as microbes are easy to manipulate to obtain enzymes of desired characteristics. However fungus is preferred over bacteria for enzymes production because of its filamentous nature. Agricultural wastes byproducts have great potential for the production of enzymes. Two indigenous carbon sources namely wheat bran and rice bran were used as substrate for amylase synthesis by Aspergillus flavus through solid state fermentation. Wheat bran gave the highest (670 ± 36μmol./min/mL) and Rice bran (428 ± 24μmol./min/mL) gave the lowest enzyme activity was obtained. Optimum inoculum size of 10% (660 ± 36μmol./min/mL), incubation period 120 hours (670 ± 28 μmol./min/mL), pH 6 (680 ± 42μmol./min/mL), temperature at 30°C (660 ± 46μmol./min/mL) gave highest yield of enzyme production. The highest yield of amylase production was obtained by the addition of MgSO4 0.2% and CaCl2 0.02% respectively. The results obtained in the present study suggest that the Aspergillus flavus may act as a potent strain for industrial production of α-amylase.

129-137

Authors: J. Balasubramanian1*, T. Sravanthi2, V. Sujitha2

The aim of the present review provides the relationship between therapeutic nanoparticles and mononuclear phagocytic system. Reticuloendothelial system (RES) represents a group of cells having the ability to take up and sequester inert particles and vital dyes. This includes macrophages and macrophage precursors, specialized endothelial cell lining the sinusoids of the liver, spleen and bone marrow, and the reticular cell of the lymphatic tissue (macrophages) and of the bone morrow (fibroblast). Nanoparticles are rapidly sequestered and retained by the organs comprising of the reticuloendothelial system (RES), mainly the liver, spleen and the bone marrow. Thus, targeting of the nanoparticles to the reticuloendothelial system (RES) is much simpler than to any other organ. In the liver, the particles are mainly retained by the scavenging periportal and midzonal Kupffer cells, while the hepatocytes and liver endothelial cells may play a secondary role under special pathophysiological conditions or for special physic- chemical characteristics of particles. The clearance of conventional nanoparticles has been proposed to occur by uptake of the nanoparticles by the reticuloendothelial system (RES). The mononuclear phagocytic system uptake of nanoparticles results in their rapid removal from the blood and accumulation in tissues involved in the RES, such as the liver and spleen. Uptake by the RES usually results in irreversible sequestering of the encapsulated drug in the RES, were it can be degraded. In addition, the uptake of the nanoparticles by the RES may result in acute impairment of the mononuclear phagocytic system and toxicity. Sterically stabilized nanoparticles, such as STEALTH nanoparticles, prolong the duration of exposure of the encapsulated nanoparticles in the systemic circulation. The presence of the PEG coating on the outside of the nanoparticles does not prevent uptake by the reticuloendothelial system, but simply reduces the rate of uptake.

143-150

Authors: Sameh S. Zaghlool1*, Basim A. Shehata2, Ali A. Abo-Seif1, Hekma A. Abd El-Latif3

Gastric ulcer is one of the most serious diseases over wide the world. There are many drugs used for the treatment of gastric ulcer, but most of these produce several adverse reactions. This study aims to investigate the protective effects of extracts of Ginger and Marshmallow on pyloric ligation-induced gastric ulcer in rats. Animals were divided into 5 Groups; a normal control group, an ulcer control group, a standard treatment group receiving famotidine (20 mg/kg), and two treatment groups receiving Ginger (100 mg/kg) and Marshmallow (100 mg/kg). Treatments were given orally for 14 days. On the 15th day, animals were subjected to pyloric ligation except for the normal control group. Four hours later, rat stomachs were excised and gastric juice and blood samples were collected. Pyloric ligation caused significant elevations in ulcer number, ulcer index, gastric volume, titratble acidity, acid output, mucin content and peptic activity, accompanied by significant decreases in blood superoxide dismutase activity (SOD) activity and gastric mucosal nitric oxide (NO) and glutathione (GSH) contents. In addition, elevations in gastric mucosal lipid peroxide and histamine contents were observed. Pretreatment with famotidine, ginger or marshmallow significantly corrected all blood and tissue parameters by varying degrees. Famotidine, ginger and marshmallow may protect against pyloric ligation-induced peptic ulcer in rats, being promising for further clinical trials.

157-162

Authors: Pallavi Thakur1, Raman Chawla1*, Ankita Singh Chakotiya1, Raghav Nagpal1, Ankit Tanwar1, Rajeev Goel1, Alka Narula2, Rajesh Arora3, Rakesh K. Sharma1

The herbal extracts of Camellia sinensis (Cs) and Andrographis paniculata (Ap) with standard F/P ratio (flavonoid/ polyphenol) of 0.26-0.27 were used for evaluation of anti-microbial efficacy against Vancomycin Resistant Enterococci (VRE). The bioactivity fingerprints useful for standardizing both extracts were found to be coherent with respect to their Diphenyl picryl hydrazyl radical scavenging potential (IC50 = ~ 30µg/ml). In vitro efficacy analysis of Cs and Ap extracts using exponentially growing culture of VRE revealed Minimum Inhibitory Concentration (MIC) as 200µg/mL and 50µg/mL respectively. The comparative analysis of their Minimum Bactericidal Concentration (MBC) revealed that 5X concentration of Cs as against 10X concentration of Ap was required to achieve zero microbial count. Such analysis indicated that both extracts would be useful for future analysis. In vivo pre-clinical evaluation studies are underway for utilization of identified herbal leads for trials on humans.

168-171

Authors: Meghraj Singh Baghel*, Kavita Goswami

Cancer is referred as uncontrolled growth of abnormal cell mass. Out of the several types of cancer, prostate cancer (PC) has become a major public health problem in men worldwide. Bcl-2 and p27 proteins are important regulatory molecules of cell cycle. Failure of cell cycle regulation leads to uncontrolled cell proliferation and causes cancer. For designing an effective structural based targeted drug, the assessment of protein-protein and protein-ligand interaction is indispensable. Therefore for treatment of PC, we selected a ribosome inactivating protein, Ricin, for assessment of its therapeutic nature. In the present work through CLUSTAL-W phylogenetic analysis, we found that Bcl-2 protein was found more conserved than p27. Further Bcl-2 was selected as target molecule for docking study with Ricin protein and other chemically synthetic inhibitor molecules i.e. 2-difluoromethylornithine (DFMO) and Sarcosine, as lead molecule. Through HEX5.1 docking software docking was performed between targeted receptor and lead molecules. Energy maximum (Emax= -93.12) and energy minimum (Emin= -163.07) was observed for docking complex of optimised and energy minimised structure of Bcl-2 receptor with Ricin, which in turn shows that it is highly stable interaction. On the other hand, for synthetic inhibitors, we found energy maximum (DFMO; Emax= -77.17, Emin= -117.83 and Sarcosine; Emax= -72.23, Emin= -103.00) and energy minimum, which are significant more as compared to Ricin docking complex. Due to ricin docking complex having less energies shows stable interaction with Bcl-2. We also observed that Ricin is less toxic (lesser log P value) as compared to other molecules by toxicity analysis by ADME/TOX server. These evidences show this Ricin could be better drug for PC. Further results are needed to validate by in vitro and in vivo study to make proper elucidation of drug for better PC treatment.

176-181

Authors: T. K. Laha1*, S. Sen1, G. Mishra2

A rapid and sensitive high performance liquid chromatographic method is described for simultaneous determination of duloxetine and its major metabolites, such as 4- hydroxy duloxetine (M7), Glucuronide conjugate of 5-hydroxy-6-methoxy duloxetine (M6) and Glucuronide conjugate of dihydrodiol duloxetine (M12) in rabbit plasma. HPLC analysis was carried out on a µ-Bondapak C18 column (250 mm × 4.6 mm, 5µm particle size) using methanol: phosphate buffer (pH 7.9, 50 mM) (7:3 v/v) as the mobile phase at a flow rate of 1.5 ml/min. Detection was carried out at 224 nm with an UV detector. The above metabolites present in the rabbit plasma were characterized by retro-synthesis followed by NMR and MS study for structure confirmation and finally injected separately into the HPLC system. All the three retention time matches with the metabolites present in the plasma sample.

188-192

Authors: Ayyagari Rama Murthy1*, K. Raghu Babu1, N. A. Vekariya2

Objective of the study was to develop and validate a new rapid, sensitive, reverse phase High Performance Liquid Chromatography technique for the estimation of Olanzapine in bulk and dosage form. Chromatographic separation was achieved on a BDS Hypersil C18, (150 mm × 4.6 mm, 5μm) with a mobile phase of 0.01M Tetra butyl ammonium hydrogen sulphate: Methanol (60:40) v/v. The flow rate was 1.0 ml/min, temperature of the column was maintained at ambient and detection was made at 228 nm. The run time was as short as 10.0 min. The developed method was validated according to the International Conference on Harmonization (ICH) guidelines with respect to linearity, accuracy, precision, specificity and robustness. The developed method was linear for Olanzapine from 10 - 80μg/ml and the linear regression obtained was 0.9994. Recovery of Olanzapine was found to be in the range of 99.6 -101.7%. Precision, evaluated by intra- and inter-day assays had relative standard deviation (R.S.D) values within 1.0 %. The ruggedness of the method was validated by performing the analysis by two different analysts and no significant difference in the result was found. Robustness of the method was checked by varying flow rate by 0.2 ml/min and the result was not affected significantly. The analysis of the marketed sample indicated that the method can also be applied to dosage form successfully. The method is precise, accurate, linear, robust and fast. The retention time and run time was very short; hence it is cost effective, making it more economical and rapid. Hence, this method can be used for the analysis of large number of samples.

198-204

Authors: Jasmine Karanjia*

This paper describes two sensitive, accurate and precise chemometric spectrophotometric methods for the simultaneous determination of Cinitapride hydrogen tartarate (CNT) and Pantoprazole sodium (PANTO) in bulk powder and capsules without prior separation. Multivariate calibration chemometric methods are proposed for simultaneous determination of CNT and PANTO. The chemometric methods applied are Principal Component Regression (PCR) and Partial Least Squares (PLS). These approaches are successfully applied to quantify both drugs using the information included in the absorption spectra of appropriate solutions. In these multivariate methods, calibration sets of standard samples composed of different mixtures of CNT and PANTO have been designed. The methods were validated according to The International Conference on Harmonization (ICH) guidelines. The specificity of the proposed methods was tested using laboratory-prepared mixtures. The developed methods were successfully applied for the determination of CNT and PANTO in bulk powder and dosage form combination.

01-07

Authors: V. Tischlerová1, A. Valenčáková2*

Nanotechnology, along with other fields such as theranostics, genomics or proteomics, has one of the most potential uses in prevention, diagnosis and treatment of diseases. Formulation of different nanoparticles and modification of their surface, in general, is required to lower side effects of drugs and to improve their response in human body. The use of nanoparticles as drug carriers may improve cancer therapy and reduce harmful side effects of chemotherapy and also radiotherapy. Moreover, together with imaging contrast agents nanoparticles have great perspective in cancer diagnosis. This review focuses on chemotherapeutics that are already used and studied in combination with systems and particles in nanoscale size. The most commonly used materials for nanoparticle carriers are magnetic nanoparticles, polymer drug conjugates, dendrimers, liposomes etc. The design of nanoparticles, characterized by their material composition, size, shape, flexibility, and surface properties, essentially dictates their therapeutic outcome.

13-21

Authors: Vyshnavi. V, Indira. S*, Prathima Srinivas

Loratadine is an antihistaminic drug, used in the treatment of allergic inflammation. Poor bioavailability of the drug from conventional dosage forms is especially attributable to mucociliary clearance and transient residence time. These problems can be reduced by the employment of niosomal in situ gelling system. In situ gelling of niosomal drops was developed to maintain the drug localization for extended period of time. The niosomal in situ gel formulation was transformed into gel once it is instilled into the nasal cavity. Niosomes were formulated using various surfactants (span 20, 40, 60 and 80) in different ratios using thin film hydration technique. Niosomes were evaluated for particle size, drug entrapment efficiency and in-vitro drug release. Niosomes prepared using cholesterol and span 60 in the ratio 1:1 (F3) showed higher entrapment efficiency (94.87%) and in-vitro drug release (59.90%) was optimized. The optimized niosomes were developed into in situ gel (pH induced and thermoreversible). The gels were evaluated for gelling capability, pH, viscosity, drug content and in-vitro drug release. Ex-vivo permeation was performed for optimized in situ gels (G2 and T5). The flux (Jss) and Permeability Coefficient (Kp) was found to be higher for G2. Hence niosomal in situ gelling system may have its potential applications than the conventional nasal formulations and to improve the bioavailability of the drug through its longer residence time and ability to sustain drug release with minimal loss of drug.

27-32

Authors: V. S. Nade*, L. A. Kawale

The present study aimed to evaluate the anti-amnesic potential of ethyl acetate soluble fraction of methanolic extract of Morus alba L (EASF). The effect of EASF of Morus alba on learning and memory was evaluated by using object recognition test (ORT), elevated plus maze (EPM) and water maze test (WMT). EASF was administrated in doses of 25, 50 and 100 mg/kg p.o. Scopolamine (1 mg/kg, i.p.) was used to induce impairment in memory. The effect of EASF was also studied on serotonin–induced contractions of isolated rat fundus. EASF significantly improved discrimination index (DI) in ORT. Pretreatment with EASF significantly increased transfer latency (TL) and swimming time in target quadrant in EPM and WMT respectively. The result of the in-vitro study showed that EASF inhibited serotonin-induced contractions on rat fundus, possessing anti-serotonergic activity. Thus, the present study revealed that EASF of M. alba has significantly improved learning and memory through its anti-serotonergic mechanism. Morus alba may be useful for the treatment of dementia and other cognitive disorders.

40-45

Authors: René Kamgang1*, Michel Archange Fokam Tagne2, Hortense Gonsu Kamga3, Paul Aimé Noubissi1, Marie Christine Fonkoua4, Jean Louis Essame Oyono5

Euphorbia scordifolia is traditionally used in Cameroon for the treatment of many diseases, including diarrhea. We investigated the effect of the aqueous ethanol extract of Euphorbia scordifolia (EWES) on intestinal propulsion, bacterial in vitro growth and in vivo shigellosis. To value effects of EWES on intestinal propulsion, normal rats orally received 0, 50, or 75 mg/kg bw extract and the distance covered by the charcoal meal was measured. Shigellosis was induced by oral administration of 1.2 × 109 Shigella dysenteriae type A1 (Sd1) to the rats. Diarrheic rats were treated for 6 days with 50, 75 mg/kg EWES or 20 mg/kg norfloxacin. Nitric oxide (NO) production in blood and in colonic homogenate; blood parameters, diarrheal stool weight and Sd1 density were assessed, and death rate recorded. In vitro, the minimal inhibitory and minimal bactericidal concentrations of EWES were 4000 and 12000µg/mL respectively. EWES significantly (P<0.01) reduced intestinal transit, bacterial growth, diarrheal stool weight and decreased NO production in large intestine (P<0.01). The results suggest that Euphorbia scordifolia possesses bactericidic and antidiarrheic properties, and could be a therapeutic alternative for diarrhea of bacterial etiology.

52-58

Authors: A Kalaiselvi1*, G Aadhinath Reddy2, V Ramalingam1

Nowadays, aluminium (Al) exposure has been increasing and it has the potential to be toxic in animal and humans. In recent years, ginger has become a subject of interest because of its beneficial effects on human health. The purpose of the present study to investigate the effect of ginger extract on serum biochemical parameters of aluminium chloride (AlCl3) induced male rats. 24 Wistar rats (6 in each group) distributed into 4 groups. Control group received distilled water as vehicle; In E1 group, animal received AlCl3 orally (100 mg/kg bw), E2 group received AlCl3 (100 mg/kg bw) and simultaneously with ginger extract (50 mg/kg bw) and E3 group received ginger extract alone (50 mg/kg bw) for 60 days. At the end of the experimental period, blood samples were collected for separating the serum for biochemical analyses. The results showed that oral administration of aluminium revealed a significant increase in the levels of serum glucose, total protein, globulin, albumin, urea, uric acid, creatinine, lipid profile and serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) and no change was noted in bilirubin. The extract of ginger decreased the activities serum levels of AST, ALT, ALP, lipid profile and all the parameter studied. It was concluded that the consumption of ginger protects the liver and kidney against the Aluminium toxicity. In addition, ginger is capable of improving hyperlipidemia and the impaired kidney functions.

63-67

Authors: V. Rajalakshimi1*, K. Gauthaman1, Adel Ali Alhazzani2, R. Kamalraj3, Nazima Nisar4, G. Devdass5

The polyherbal formulation of Seenthil churanam is composition of whole plant extracts of Eclipta prostata, Tinospora cordifolia and the dried powder form of Earthworm used in folk medicine. The study was conducted to evaluate the scientific figures for the treatment of anti-inflammatory and analgesic activity of ethanolic extract of Seenthil churanam by acetic acid induced writhing test and eddy’s hot plate method, and carrageenan induced paw edema method. There was significant response in analgesic and inflammatory activity at high dose (400 mg/kg) compared to low dose 200 mg/kg against the standards Analgin (500 mg/kg), Aspirin (100 mg/kg) and Diclofenac sodium (100 mg/kg) body weight of mice and rats. The results of this study show that the chronic oral administration of an ethanolic extract of Seenthil churanam at a 400 mg/kg body weight dosage be a good alternative natural medicine for analgesics and anti-inflammatory drug without side effects.

73-77

Authors: N. Sharief Mohammad1, A. Srinivasulu1, B.Chittibabu2, Uma Maheswara Rao V3*

Avicennia officinalis (AO) belongs to family Avicenniaceae. It finds a prominent place in folk medicine. Hence the present work was carried out for the identification of bioactive principles in the fruits of Avicennia officinalis and screened for the antibacterial activity against selected test cultures. The methanolic extracts has high efficacy against the test cultures. The methanolic extracts were further purified by column chromatography. With the aid of 1H NMR, 13C NMR and GC-MS analysis. The structure of the compounds were detected as Phenols namely 1, 2, 3 Benzene triol and 4, 4’-(1-methylethyldiene)bis2-methyl and both the compounds exhibited antibacterial activity. This report of ours is the first one regarding the 1, 2, 3 Benzene triol and 4, 4’-(1-methylethyldiene) bis 2-methyl in the plant Avicennia officinalis.

83-88

Authors: Ekta Sharma, Bhupinder Singh Arora*

In the present study, with the appliance of hyphenated techniques involving high performance liquid chromatography with diode array detection directly coupled with 1H nuclear magnetic resonance spectroscopy (LC-NMR) and electrospray ionization mass spectrometry (LC-ESI-MS), four known aryltetrahydronephthalene lignans (podophyllotoxin 4-O-glucopyranoside, 4’-demethylpodophyllotoxin, podophyllotxin and dehydropodophyllotoxin) have been well characterized from methanol extract of rhizomes of Podophyllum hexandrum Royle without involving the time consuming steps of isolation and purification of individual constituents of the extract. Also, a simple, sensitive and reproducible LC-MS method has been developed for the quantitative analysis of podophyllotoxin in crude methanolic extract from different parts of Podophyllum hexandrum Royle. The present work adds up to the existing significance of hyphenated techniques combining chromatographic technologies with spectroscopic techniques for both qualitative and quantitative analysis of natural product extracts or fractions without any need of reference standards for the markers.

96-99

Authors: K.P Thomford1*, M.L.K Mensah2, R.A Dickson3, M. Sakyiamah1 D.A Edoh1, K. Annan2

Quantitative chemical fingerprinting of herbal medicines provides an avenue for addressing the challenge with their quality. In this report, chemical markers that could be labelled as active markers in the polyherbal product and its component raw materials were assayed using a simultaneous RP-HPLC method. These markers were rutin (RU), quercetin (QE) and kaempferol (KA) which are well known for their antimicrobial activities. The quantitative assay of the markers using a validated HPLC method showed the herbal product (EAF-2011) contained 8.6810% (w/w) of rutin, 0.2670% (w/w) of quercetin and 0.0610% (w/w) of kaempferol. At least two (2) of these flavonoids were detected in each of the component raw plant materials: Psidium guajava [(RU-8.5860% (w/w); QE-0.5430% (w/w); KA - 0.1003% (w/w)], Alchornea cordifolia [RU-2.5540% (w/w); QE-0.0536% (w/w)], Zanthoxylum zanthoxyloides [RU-5.8060 % (w/w); KA-0.0174% (w/w)], Tridax procumbens [RU-2.4390% (w/w); QE-0.1040% (w/w); KA-0.0510% (w/w)] and Eugenia caryophyllata [QE-0.4080% (w/w); KA-0.0771% (w/w)]. The results obtained indicate that the HPLC method, selected chemical markers and chromatograms produced are ideal for the quality control of the product and its component raw materials.

105-109

Authors: Suman Pattanayak*, Ash Rani. Y

The present paper describes a reverse phase HPLC method for simultaneous estimation of Trifluoperazine and Isopropamide from their combined tablet dosage form. The proposed RP-HPLC method utilizes an Agilent Zorbax, C18 column (250 × 4.6 mm, 5μm particle size) using a mobile phase consisting of mixture of Buffer: Acetonitrile (80: 20 v/v) mobile phase flow rate of 0.8 ml/min; and UV detection at 227nm. The retention time for Trifluoperazine and Isopropamide were 2.4 and 3.62 minutes respectively. The method was linear in the range of 40-120μg/ml and 100-300μg/ml and correlation coefficients were 1.00 and 1.00 for Trifluoperazine and Isopropamide respectively. The limit of detection was found to be 2.963 and 2.985µg/ml for Trifluoperazine and Isopropamide respectively. Limit of quantification was found to be 9.877 and 9.9502 for Trifluoperazine and Isopropamide respectively. The percentage recoveries for Trifluoperazine and Isopropamide ranged from 100.03-100.36% and 100.00-100.07 % respectively. The percentage of RSD for precision of the method was found to be less than 2%. The proposed method can be used for the routine analysis for the estimation of these drugs in combined dosage form.

116-119

Authors: Kshitija Kardile, Mrinalini C. Damle*

The objective of this study was to develop stability UV method for estimation of Dronedarone Hydrochloride in tablet to compare % degradation of this drug determined by this UV method to HPTLC method reported by us earlier. Analysis of bulk as well as marketed tablet formulation by exposure to same degradative conditions was done and % degradation obtained by spectroscopy was determined. Method development and validation was done as per ICH guidelines. % degradation observed by both methods was comparable. The variation in the results of this drug estimated by HPTLC and spectrophotometric method was statistically insignificant. This suggests that the spectrophotometric method is suitable to determine stability profile. Hence a simple UV spectrophotometry can be used to monitor stability of Dronedarone Hydrochloride.

123-125

Authors: M. Neagu1*, G. Soceanu1, A. Bucur1, A. Tanase2

In the present study, an analytical HPLC method was developed for the estimation of Fipronil, Pyriproxyfen, Benzyl alcohol, Butylhydroxyanisole (BHA) and Butylhydroxytoluene (BHT) and related substances from a veterinary pharmaceutical product. The method showed linearity in the range of 0.44-200% with a correlation coefficient of 0.99. The method was validated for different validation parameters such as: specificity (peak identification and interference study), precision, accuracy, linearity, robustness, limit of detection (LOD) and limit of quantitation (LOQ), stability of standard solutions and sample solution. The results were found to be within the acceptance limits as per ICH guidelines.

253-262

Authors: Ahsas Goyal*, Neetu Agrawal, Bhupesh C. Semwal, Yogesh Murti

Hutchinson-Gilford Progeria Syndrome (HGPS) is an extremely rare genetic disorder characterized by premature aging, involving aberrant splicing of the LMNA gene, resulting in the production of a disease-causing mutant lamin A protein called progerin. Clinical manifestations are evident by the first or second year of life and include the physical characteristics usually associated with the elderly. Because neither parent carries or expresses the mutation, each case is believed to represent a sporadic, new mutation that happens most notably in a single sperm or egg immediately prior to conception. Clinical trials investigating farnesyltransferase inhibitors (FTIs), statins, and bisphosphonates as HGPS treatments are currently underway. FTIs prevent farnesylation and localization of progerin to the cell membrane but do not repair the function of the abnormal progerin protein within the cytoplasm that may result in abnormalities in cell function and DNA repair that, therefore, would not be treated with these drugs. Thus some other novel treatment strategies are required for the more effective treatment. This review summarizes the clinical characteristics of this disease, the underlying mutation in the lamin A (LMNA) gene that results in this phenotype and the recent advances in treatment strategies.

271-277

Authors: Sushma Gupta*, Tania Munjal, Amanjot Kaur, Kamaljeet S Bagga

Cefopodoxime Proxetil belongs to BCS class IV and used in treatment of upper respiratory tract and urinary tract infections. Solid dispersions (SDs) are one of the most promising strategies to improve the solubility, dissolution and ultimately oral bioavailability of such poorly water soluble drugs. The main objective of the present research was to formulate Cefopodoxime Proxetil solid dispersion employing two methods namely hot melt granulation and solvent evaporation method. The PEG 4000 and PEG 6000 were used as carrier in varied proportion (1:1, 1:2, 1:3 and 1:4 w/w). Results of FT-IR spectra revealed no potential chemical incompatibility between drug and excipients. Enhancement in the percent drug released and dissolution rate was observed in SD of PEG 6000 as to PEG 4000 and pure drug. Drug release kinetics studies revealed that the drug release from the formulations followed non-fickian diffusion and the best fitted model for drug release for Korsmeyer Peppas Model. No sharp peaks were observed in both solid dispersions (comprising PEG 4000 & PEG 6000) in PXRD spectra revealing the formation of amorphous product. Similar results were observed in DSC studies indicating disappearance of sharp fusion endothermic peak i.e. conversion of crystalline form into amorphous form. These results were further supported by SEM studies showing disappearance of crystal habit in these formulations.

283-290

Authors: Usha Rani N*, Nagoor Ankani

A stability indicating, accurate, specific, linear and sensitive reverse phase-HPLC method was developed and validated for the determination of Rosuvastatin calcium (ROS) in pharmaceutical dosage form. The chromatographic separation was performed using Eclipse XDB plus C18 Column (4.6 × 150 mm, 5μm particle size). Mobile phase was made by using Acetonitrile: Water (60:40 v/v) at a flow rate of 1.0 ml/min with the detection wavelength at 242 nm. The retention time of ROS was found to be about 1.787 min. The linearity was obtained in the concentration range of 10-50 μg/ml with a correlation coefficient of 0.999. The percentage purity of ROS was found to be 100.06 %. The percentage recovery was determined for ROS and found to be 100.53 %. The developed analytical method was validated for linearity, precision, accuracy, ruggedness, robustness, specificity and system suitability which were within the acceptance limit according to ICH guidelines. All the degradation products obtained by stress conditions were found to be well separated from the principal peak, which means that the ROS peaks were highly pure in all chromatograms obtained. The developed method was found to be applicable for routine quality control and stability analysis of ROS in pharmaceutical dosage forms.

303--309

Authors: Vineeta Khanvilkar*, Leena Patil, Vilasrao Kadam

Standardization of herbal formulation is essential in order to assess the quality, purity, safety and efficacy of drugs. World health organization (WHO) in 1999 has given a detail protocol for the standardization of herbal drugs comprising of a single content but very little literature is available for the standardization of polyherbal formulations. Chitrakadi Vati is official in ayurvedic formulary of India and it is prescribed for the treatment of irritable bowel syndrome, rheumatoid arthritis and loss of appetite. In the present work, attempt has been made to develop a chromatographic method for standardization of Chitrakadi Vati. All raw materials used were standardized by macroscopic, microscopic and physico-chemical parameters. Piperine in Piper nigrum and Piper longum; Plumbagin in Plumbago zeylanica are active components in the formulation and can be considered as marker compounds. A simple, rapid, precise, accurate and reproducible High Performance Thin Layer Chromatography (HPTLC) densitometric method was developed. The separation was performed on TLC aluminium plates precoated with silica gel 60 F254, good separation was achieved in the mobile phase of toluene: ethyl acetate: formic acid (7.5:2.5:0.5 v/v/v) and densitometry determination of piperine (0.39 ± 0.02) and plumbagin (0.70 ± 0.02) was carried out at single wavelength scanning at 280 nm. The developed method has been validated as per ICH (International conference on harmonization) guidelines. The developed and validated HPTLC method was applied for standardization of Chitrakadi Vati by detection and quantification of markers piperine and plumbagin simultaneously from in-house and marketed formulations.

323-328

Authors: AT Tchinda1*, GA Agbor1, DE Tsala2, AJG Yaya1, EN Nga3, E Talla4, J-N Wauters5, M Frédérich5

The present study evaluates the antioxidant activity of the different flavonoids of the fruits of Xylopia parviflora used in Cameroon as spice in common traditional dishes. The fruits were successively extracted with hexane and methanol. The isolation of flavonoids was guided by the DPPH-TLC technique. The methanol crude extract and isolated compounds were screened for antioxidant and free radical scavenging activities using DPPH radical-scavenging, β-carotene/linoleic acid and ferric reducing antioxidant power (FRAP) assays. The total phenolic content of the methanol crude extract was determined by Folin–Ciocalteu method. The DPPH-TLC technique led to the isolation of (+)-catechin (1), kaempferol 3-O-arabinofuranoside (2) and quercetin 3-O-arabinofuranoside (3) identified by NMR and mass spectra analysis. In the colorimetric DPPH test, compound 1 had the same activity (EC50 8.1µg/ml) as butylated hydroxytoluene (BHT) used as standard while compound 3 and the methanol crude extract were less active (EC50 17.2µg/ml). Compound 2 was completely inactive. The total phenolic content of the fruit extracts was 113.03 mg gallic acid equivalents per g of extract. In the carotene bleaching test at the highest concentration of 100µg/mL, the order of inhibition of β-carotene discoloration was BHT > crude extract > quercetin 3-O-arabinofuranoside (3) > catechin (1) > kaempferol 3-O- arabinofuranoside (2). In the reducing power assay, compound 3 was more active at concentrations 40-100µg/mL. The HPLC analysis of the methanol crude extract revealed the presence of compounds 1-3 and unidentified phenolic compounds. The antioxidant activity of the methanol extract is probably due to the presence of compounds 1 and 3.

334-340

Authors: Ajithadas Aruna*, Nandhini SR, Karthikeyan V, Bose P

To compare in-vitro antioxidant activity of Methanolic extract of Costus pictus (MECP) and its Silver nanoparticles (MECPAgNPs) by various methods. Preliminary phytochemical screening of MECP was done by standard procedure. Synthesis of silver nanoparticles from MECP was done. In-vitro anti-oxidant activity of the MECP & MECPAgNPs were studied by DPPH assay, H2O2 scavenging activity, Phosphomolybdenum Method, FRAP and reducing power assay. The total Phenolic content, Flavonoid content & Vitamin C were estimated by using Gallic acid, Quercetin and standard Ascorbic acid calibration curve respectively. Preliminary phytochemical screening showed the presence of carbohydrates, triterpenoids, proteins, alkaloids, tannins, saponins, flavonoids, sterols and volatile oil. In-vitro antioxidant methods were resulted, the extract and the nanoparticles showed a dose dependent reducing ability. The nanoparticles at the same concentration offered much better activity than the extract alone. Phenolic content, Flavonoid content and Vitamin C amount of the MECPAgNPs was higher than MECP. These determination and quantification gives the information about the amount of secondary metabolites present in the MECPAgNPs was higher than the MECP which is responsible for the therapeutic or pharmacological activity of the plant. The MECPAgNPs showed very potent anti oxidant activity as compared to MECP.

345-347

Authors: Kalpana Devi Rajesh1*, Nakulan Valsala Rajesh2, Subramani Vasantha1, Solomon Jeeva3

Ethanolic, aqueous and Petroleum ether extracts from the whole fern, Oleandra musifolia (Bl.) Presl. (Family-Oleandraceae) was investigated for anthelmintic activity against Haemonchus contortus, with varied drug concentration (25, 50, and 100) mg/ml and the time taken for paralysis and death of the worm was determined. The in-vitro anthelmintic activity was compared with standard reference drug Albendazole and control distilled water. The result showed ethanolic extract performed well to express the anthelmintic efficacy than aqueous and petroleum ether extracts and the standard Albendazole. However efficacy is better and directly proportional to the increase in varied drug concentration. The efficacy is determined as lesser time taken to cause paralysis and death of Haemonchus contortus as compared to standard reference drug.

351-355

Authors: Kirti Rani*, Vartika Mehta

In present work, Cicer arietinum amylase was encapsulated by emulsification through covalent coupling by glutaraldehyde into chemically modified bovine serum albumin. Biodegradation of coconut oil driven emulsified bovine serum albumin encapsulated Cicer arietinum amylase was carried out by the alkaline protease for its controlled and sustained release of encapsulated enzyme from prepared microparticles of encapsulated Cicer arietinum amylase and its stability increased up to 6 months as compared to free enzyme. Its biodegradation was carried out by the using different concentration of alkaline protease (5U, 10U, 15U, 20U, 25U, 30U, 35U, 40U). Further, this coconut oil driven chemically modified bovine serum albumin microparticles of encapsulated Cicer arietinum amylase with alkaline protease were used with detergents for washing of stained cloths which have rust, gel pen ink, grease and chocolate strains. These chosen strains are very commonly present on uniforms of school going children which are very tough upon drying, hence, not to be easily vanish with well known brand detergents upon in one wash. But, the mixture solution of coconut oil driven chemically modified bovine serum albumin microparticles of encapsulated Cicer arietinum amylase along with alkaline protease were used with detergents powder for washing of these dry tough strains (rust, gel pen ink, grease and chocolate strains) leads to vanishing these strains very fast with absolute clear results were found as compared to results of washing of stained cloths with detergents only.