Open Access Original Research Article

Antifungal Properties and Chemical Analysis of Essential Oil from Vitex negundo Seeds

Hong-Wei Ai, Yue-Xin Kang, Yang Cao, Cheng-Jian Zheng

Journal of Pharmaceutical Research International, Page 541-548
DOI: 10.9734/BJPR/2014/7079

Aims: To investigate the constituents and antimicrobial activity of essential oil from Vitex negundo seeds.
Study Design: The essential oil of Vitex negundo seeds was prepared by hydrodistillation. GC/MS technique was used to determine the volatile constituents in the oil. A total of 13 isolates of bacteria and fungi were employed to evaluate the antimicrobial activity of the oil.
Place and Duration of Study: Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, between April 2010 and September 2010.
Methodology: Essential oil from Vitex negundo seeds were obtained by hydrodistillation, with constituents analyzed by GC-MS. Antimicrobial activity of the oil was evaluated against both Gram-positive and Gram-negative bacteria and fungi.
Results: A total of forty-two components, representing 91.36% of the oil, were identified. n-Hexadecanoic acid (17.68%), eudesm-4(14)-en-11-ol (12.39%) and caryophyllene oxide (10.79%) were found to be the major constituents. The essential oil exhibited significant antifungal activity against Candida albicans at MIC 4.0 μg/ml.
Conclusion: The essential oil of Vitex negundo seeds showed potential antifungal activity and can be of potential use in pharmaceutical fields.

Open Access Original Research Article

Solubilization of Poorly Water Soluble Drug Using Mixed Solvency Approach for Aqueous Injection

Love Kumar Soni, Shailendra Singh Solanki, Rajesh Kumar Maheshwari

Journal of Pharmaceutical Research International, Page 549-568
DOI: 10.9734/BJPR/2014/4281

Aim: 1) To increase water solubility of indomethacin drug using mixed solvency approach.
2) To employ use of non toxic solubilizers for increasing solubility of a poorly water soluble drug.
Study Design: Trial and error based experimental study.
Place and Duration of Study: School of Pharmacy, Devi Ahilya Vishwavidyalaya, UTD, Takshashila Campus, Indore, India and College of Pharmacy, IPS Academy, Indore, India between Jan. 2011 and June. 2012.
Methodology: By making mixed solvent (40%) blends of selected water soluble substances from the hydrotropes (urea, sodium benzoate, sodium citrate, nicotinamide); water-soluble solids (PEG- 4000, PEG-6000); and co-solvents (propylene glycol, glycerine, PEG-200, PEG-400, PEG 600) solubility studies were performed with indomethacin (model drug). On the basis of solubility studies formulation was developed. The solubilized drug and prepared formulation were characterized by ultraviolet and infrared techniques. Various properties of solution such as pH, viscosity, specific gravity and surface tension were studied. The developed formulation was studied for physical and chemical stability.
Result: Aqueous solubility of drug in case of selected blends ranged from 14.55 mg/ml – 19.96 mg/ml (as compared to the solubility in distilled water 0.0464 ± 0.007 mg/ml). The enhancement in the solubility of drug in a mixed solvent containing 10% sodium benzoate, 5% sodium citrate and 25 % S cosolvent (25% S cosolvent contains PEG200, PEG 400, PEG600, Glycerine and Propylene glycol) was more than 400 fold. Prepared formulation F10 show appreciable physical and chemical stability.
Conclusion: The results of this study provide guidance for developing an injectable product and strategies for improving solubility as well as stability of poorly water soluble drug using the concept of mixed solvency. The proposed technique is economical, convenient and safe. The application of mixed solvency approach in the development of formulations shall prove a boon for pharmaceutical industries.

Open Access Original Research Article

Phytochemical Screening and Antipyretic Effect of Curcuma zedoaria Rosc. (Zingiberaceae) Rhizome

Md. Golam Azam, Md. Shibly Noman, Md. Mamun Al-Amin

Journal of Pharmaceutical Research International, Page 569-575
DOI: 10.9734/BJPR/2014/7225

Aims: To investigate the antipyretic effect of ethanol extract of Curcuma zedoaria Rosc. (Zingiberaceae) rhizome in animal model.
Study Design: Extraction of plant constituents and evaluation of elevated body temperature lowering activity.
Place and Duration of Study: Department of Pharmacy, North South University, Dhaka between October 2012 and August 2013.
Methodology: We have performed phytochemical screening and evaluated antipyretic activity of ethanol extract of the rhizome of Curcuma zedoaria by yeast-induced pyresis method. Ethanol extract of Curcuma zedoaria rhizome was administered to healthy rats.
Results: The results showed that the ethanol extract of Curcuma zedoaria significantly reduced yeast-induced elevated body temperature in rats in a dose dependent manner and the antipyretic effect at a dose of 750 mg/kg was comparable to that of the standard antipyretic drug paracetamol (10 mg/kg). Phytochemical screening of ethanol extract showed presence of tannins, flavonoids, saponins, alkaloids, terpinoids, carbohydrates and steroids as main constituents in Curcuma zedoaria extract some of which may possess antipyretic activity.
Conclusion: The results justify the traditional use of the ethanol extract of Curcuma zedoaria in the treatment of fever.

Open Access Original Research Article

Antidepressant-like Activity of Celastrus paniculatus Seed Oil in Mice Subjected to Chronic Unpredictable Mild Stress

Rekha Valecha, Dinesh Dhingra

Journal of Pharmaceutical Research International, Page 576-593
DOI: 10.9734/BJPR/2014/5677

Aims: The present study was done to evaluate the antidepressant-like effect of Celastrus paniculatus seed oil in Swiss young albino mice subjected to chronic unpredictable mild stress.
Study Design: Prospective.
Place and Duration of Study: Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science and Technology, Hisar, Haryana, India, between August 2012 to February 2013.
Methodology: The behavioral models such as forced swim test (FST) and sucrose preference test were used to evaluate the effect of seed oil on depression in mice. The oil (50, 100 and 200 mg/kg, p.o.) and fluoxetine (20 mg/kg, p.o.) per se were administered for 14 successive days to unstressed and stressed mice. The effect of oil on locomotor activity of mice was also evaluated. In addition, the effects of oil on brain monoamine oxidase-A, malondialdehyde levels, reduced glutathione and catalase activities; and plasma corticosterone and nitrite levels were also assessed.
Results: The oil significantly decreased immobility period of both unstressed and stressed mice in FST as compared to the control. The oil also prevented the stress-induced decrease in sucrose preference, indicating significant antidepressant-like activity. There was no significant effect on locomotor activity of mice by the oil. Antidepressant-like activity of the oil was comparable to the standard drug, fluoxetine. The oil also significantly decreased monoamine oxidase-A (MAO-A) activity, malondialdehyde levels in both unstressed and stressed mice; and significantly prevented stress-induced decrease in reduced glutathione and catalase activities. It also significantly attenuated stress induced-increase in plasma nitrite and corticosterone levels.
Conclusion: Celastrus paniculatus seed oil showed significant antidepressant-like activity in both unstressed and chronic unpredictable mild stressed mice probably due to inhibition of MAO-A activity, decrease in plasma nitrite levels; and through scavenging of free radicals. In addition, the oil also showed antidepressant-like activity in stressed mice probably through decrease in plasma corticosterone levels.

Open Access Original Research Article

Antiplasmodial Efficacy of Stem Bark Extracts of Pseudocedrela kotschyi in Mice Infected with Plasmodium berghei berghei

A. Dawet, D. P. Yakubu

Journal of Pharmaceutical Research International, Page 594-607
DOI: 10.9734/BJPR/2014/2809

Aims: To evaluate the antiplasmodial efficacy of stem bark extracts of P. kotschyi against P. berghei berghei in mice.
Study Design: Extraction and administration of plant extracts and evaluation of daily parasitaemia of infected mice.
Place and Duration of Study: Department of Pharmacognosy; Animal House. Department of Pharmacology and Department of Zoology, University of Jos, Jos, Nigeria, between June, 2008 and November, 2011.
Methodology: Dry zone cedar (Pseudocedrela kotschyi) Family: Meliaceae, stem bark powder was successively extracted using ethyl acetate, ethanol and aqueous solvents. A total of one hundred and twenty mice for each extract (ethanol, ethyl acetate and aqueous) were divided into three groups of forty mice per each test (suppressive, curative and prophylactic). They were inoculated with drug sensitive NK 65 Plasmodium berghei berghei. In each test animals were divided into five groups, each consisted of eight animals and treated separately with one of the following: 50, 100, and 200 mg/kg extracts, chloroquine / pyrimethamine and normal saline. Blood films were prepared and examined, and the changes in percentage parasitaemia were evaluated.
Results: The ethanol, ethyl acetate and aqueous crude extracts of P. kotschyi at 200 mg/kg significantly (P=.05) inhibited the parasitaemia by 39.43%, 26.99% and 28.36% respectively in the suppressive test. Ethanol and ethyl acetate crude extracts also showed significant (p=.05) cure rate of 29.17 % and 20.28 % respectively. However there was no significant (p>.05) reduction in parasitaemia load in the prophylactic tests.
Conclusion: The results of the study showed that P. kotschyi stem bark indeed has antiplasmodial property.

Open Access Original Research Article

Angiogenesis and Antioxidant Activity of in vitro and in vivo Tuber of Ceropegia pusilla Wight and Arn

R. Prabakaran, K. Kalimuthu, C. Vani, C. Brindha

Journal of Pharmaceutical Research International, Page 608-616
DOI: 10.9734/BJPR/2014/7481

Background: Angiogenesis refers to the formation of new blood vessels. This biological process lays an important role in embryonic development. Ceropegia pusilla (Asclepiadaceae) used in folk medicine system to treat various health problems. Present work deals with the analysis of antioxidant and antiangiogenic properties of in vivo and in vitro tuber extracts of C. pusilla by Chicken Egg Chorioallantoic Membrane Assay (CAM) in vivo.
Methods: C. pusilla tubers of both in vivo and in vitro were extracted with ethanol and methanol. In vivo anti angiogenesis effect was studied by Chicken Egg Chorioallantoic Membrane (CAM) assay. The antioxidant properties were analysed using 1, -diphenyl-2-picryl- hydrazyl free radical scavenging assay and reducing power activity.
Result: Ethanolic extract of both in vivo and in vitro tuber showed higher angiogenic activities 58.3±13.6 and 43.2±12.2 respectively than the methanolic extract. The average number of vessels in in vivo and in vitro ethanolic extracts treated CAM was 6.48 and 5.43. In vivo and in vitro extracts exhibits comparable DPPH radical scavenging activity and better reducing power activity.
Conclusion: C. pusilla tuber extracts of both in vivo and in vitro inhibited angiogenesis by blocking normal vascularisation in chick embryo and also having better DPPH and reducing power activity.

Open Access Original Research Article

Differential Spectrophotometric Method for Determination of Cefquinome Sulphate

S. Shantier, E. Gadkariem

Journal of Pharmaceutical Research International, Page 617-625
DOI: 10.9734/BJPR/2014/5734

Differential first derivative (ΔD1) method is developed for the quantitative analysis of cefquinome sulphate (CS) in bulk and dosage form. The method is based on measuring the ΔD1 of CS in alkaline solutions against its neutral aqueous solutions as blanks. The proposed method is sensitive and highly specific, since the interference of the excipients or impurities is nullified. Beer’s law was obeyed over the concentration range 4-20 µg/mL with a limit of detection and limit of quantification 1.5 µg/mL and 4.8 µg/mL respectively.

Open Access Original Research Article

In vitro Antioxidant Activity of Lectin from Different Endophytic Fungi of Viscum album L.

T. S. Sadananda, M. Govindappa, Y. L. Ramachandra

Journal of Pharmaceutical Research International, Page 626-643
DOI: 10.9734/BJPR/2014/6702

Objective: To evaluate the antioxidant potential of lectin from the endophytic fungi of Viscum album.
Place and duration of study: Department of Biotechnology, Shridevi Institute of Engineering and Technology campus, Tumkur, Karnataka, India between November 2012 and March 2013.
Methodology: Isolation, identification of endophytic fungi from Viscum album, mass production, lectin isolation, purification using affinity chromatography. In vitro antioxidant activity of lectin using total antioxidant assay, DPPH, FRAP, Fe3+ reducing assay and H2O2 scavenging activity.
Result: Different endophytic fungi were isolated and identified as Aspergillus flavus, Fusarium oxysporum, Fusarium moniliforme, Trichothecium sp. Isolated lectin was capable of agglutinating A+ve erythrocytes, various in vitro antioxidant assays were carried out. Total antioxidant activity showed that (418µg/mg) of ascorbic acid equivalent in the Viscum album extract, among endophyhtes higher activity observed in Fusarium oxysporum (348.54 µg/mg). DPPH assay showed endophytic lectin significantly reduced DPPH free radicals with the low IC50 value observed in Aspergillus flavus (127.9µg/ml) in compared to Viscum album (92.4µg/ml). Highest FRAP value was observed in the location from Viscum album of (180.12µg/ml) among endophytes Aspergillus flavus showed (133.22µg/ml) of the ascorbic acid equivalent of Fe2+/ mg of listing. In Fe3+ reducing ability showed concentration dependent reduction in Fe3+ to Fe2+, low IC50 value was observed in endophytic lectin Fusarium oxysporum (143.4 µg/ml). These values are slightly higher in relation to Viscum album (120.3µg/ml). In H2O2 scavenging activity there was concentration dependent scavenging activity was observed, IC50 value in Fusarium moniliforme was (171.2 µg/ml), Viscum album is listing showed (183.4 µg/ml).
Conclusion: Lectin isolated from endophytic fungi relieved that it was capable of scavenging the free radicals, antioxidant activity was comparatively higher in endophytes than compared with Viscum album. Endophytic lectin could be a novel source for scavenging the free radicals having pharmaceutical value.

Open Access Original Research Article

In vitro and in vivo Antitrypanosomal Effects of Methanol and Aqueous Extracts of Picralima nitida

A. C. Ene, N. G. Edeh, C. Bonny-Okoli, O. A. Ojiako, C. O. Ujowundu, C. U. Igwe

Journal of Pharmaceutical Research International, Page 644-653
DOI: 10.9734/BJPR/2014/6628

The methanol and aqueous extracts of the leaves, fruits, seeds, stem bark and roots of Picralima nitida were studied in vitro and in vivo for activity against Trypanosoma brucei brucei in Swiss albino mice. Phytochemicals studies were also conducted for all the plant extracts The methanol extracts showed appreciably high in vitro and in vivo antitrypanosomal activities compared to the aqueous extracts of the plant. The methanol extract of the root exhibited the highest in vitro antitrypanosomal activity followed by the methanol extract of seed of Picralima nitida. Motility of Trypanosoma brucei brucei was stopped by the methanol extract of the root after 10 min, while the methanol extract of the seed of Picralima nitida stopped the motility of Trypanosoma brucei brucei at 15 min. The methanol extract of the root of Picralima nitida showed the highest in vivo antitrypanosomal activity at 100 mg/kg body weight. The extract cleared the parasite completely from the T. brucei brucei infected Swiss albino mice after day 3 of treatment. There was a statistically significant difference (p<0.05) when the level of parasitemia of the animals treated with the methanol extract of the root of Picralima nitida were compared with the other treatment groups and the untreated control. The phytochemicals detected in these extracts are tannins, flavonoids, alkaloids, steroids, terpenoids, saponins and cyanide glycosides. The in vitro and in vivo antitrypanosomal activity exhibited by these extracts might be attributed to these phytochemicals.

Open Access Original Research Article

Novel Pharmaceutical Gels Containing Glyccerihizic Acid Ammonium Salt for Chronic Wounds

Usama F. Aly, Heba F. Mansour

Journal of Pharmaceutical Research International, Page 654-668
DOI: 10.9734/BJPR/2014/7591

Aims: This study aimed at preparing pharmaceutical gels containing Glyccerihizic Acid Ammonium Salt (GAM) for healing of chronic wounds followed by in-vitro and in-vivo characterization of the prepared gels to ensure their suitability, efficacy and safety.
Place and Duration of Study: Department of Pharmaceutics, faculty of Pharmacy (El-Minia University) and Department of Histology, Faculty of medicine, El-Minia University, from September 2011 to June 2012.
Methodology: Preformulation studies including infra-red and differential scanning calorimetry were performed on the drug and the polymers to confirm their physicochemical compatibility. Pluronic F-127, Carbomer 940, sodium alginate and hydroxypropyl methyl cellulose were used as gelling agents. The gel formulations were evaluated for in-vitro release and characterized for their physicochemical properties. The base that exhibited the highest release rate was subjected to further histological studies in wounded rats to demonstrate the effect of drug concentration on the histopathological changes post applications.
Results: Physicochemical compatibility studies demonstrated the absence of interaction between the drug and the polymers. The physical appearance of the prepared gel was accepted. Microscopic examinations confirmed the absence of any particles or grittiness. Sodium alginate-based gels exhibited the highest drug release rate. Statistical analysis showed high significant differences as compared to other formulations (P<0.001). Kinetic studies showed that all formulations exhibited first-order release rate. Histopathological studies demonstrated that wound closure was faster in animals treated with 3% (w/w) drug as compared to those treated with 1% (w/w) drug. No signs of irritation were noticed among all treated animals
Conclusion: GAM prepared in sodium alginate base gave the highest in-vitro release pattern, which followed first-order release rate. A concentration of 3% w/w GAM of this base would give better healing effect in a shorter time.