Open Access Original Research Article

Simultaneous Quantification of Mesalamine and Its Metabolite N-Acetyl Mesalamine in Human Plasma by LC-MS/MS and Its Application to a Bioequivalence Study

Kanchanamala Kanala, Nagiat T. Hwisa, Babu Rao Chandu, Fathi H. Assaleh, K. Mukkanti, Prakash Katakam, Bala Sekhara Reddy Challa

Journal of Pharmaceutical Research International, Page 1568-1590
DOI: 10.9734/BJPR/2014/5350

Liquid chromatography–tandem mass spectrometry (LC–MS/MS) was used for simultaneous quantification of mesalamine and its metabolite N-acetyl mesalamine in human plasma with N-acetyl mesalamine D3 as an internal standard (IS). Chromatographic separation was performed on a Thermo, HyPURITY C18 (150 x 4.6 mm, 5 ï­m) column with an isocratic mobile phase composed of 10 mM ammonium acetate and methanol in the ratio of 85:15 (%v/v), at the flowrate of 0.6 mL/min. The drug, metabolite and internal standard were extracted by liquid-liquid extraction. The method was validated over a linear concentration range of 2-1500 ng/mL for mesalamine and 10-2000 ng/ml for N-acetyl mesalamine, which demonstrated intra and inter-day precision ranging from 1.60 to 8.63% and 2.14 to 8.67% for mesalamine and 0.99 to 5.67% and 1.72 to 4.89% for N-acetyl mesalamine respectively. Similarly, the intra- and inter-day accuracy varied from 102.70 to 105.48% and 100.64 to 103.87% for mesalamine, 99.64 to 106.22% and 100.71 to 104.27% for N-acetyl mesalamine respectively. Both analytes were found to be stable throughout freeze–thawing cycles, bench top and postoperative stability studies. The method was successfully applied to support a bioequivalance study of healthy subjects.

Open Access Original Research Article

Biosynthesis of Silver Nanoparticles Using Callus Extract of Catharanthus roseus var. alba and Assessment of Its Antimicrobial Activity

Md. Abul Barkat, Mohd. Mujeeb, Md. Samim, Shashikant Verma

Journal of Pharmaceutical Research International, Page 1591-1603
DOI: 10.9734/BJPR/2014/7785

Objective: To express a cost efficacious and environment friendly method for the green synthesis of silver nanoparticles (AgNPs) from Catharanthus roseus var. alba (C. roseus var. alba) callus extract.
Methodology: The aqueous solution of sliver nitrate containing Ag+ ions (1 mM) are integrating 100 μL of aqueous extract of callus of C. roseus var. alba and 10 mL of 1% w/v aqueous solution of polyethylene glycol 4000 to 90 mL. This was then alkalized with 0.1 NaOH (20 μL) and treated in a microwave oven (800 W) for 40 sec for the reduction of metal ion and the reaction takes place at room temperature (250C). The reduction of the Ag+ ions by aqueous callus extract of C. roseus var. alba in the solutions was monitored by UV–Visible spectrum and further characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM) and x-ray diffraction (XRD). Antibacterial activity was assessed on bacterial strain of Staphylococcus aureus and Pseudomonas aeruginosa (S. aureus and P. aeruginosa) by using the disc-diffusion assay method.
Results: Characterization of AgNPs was done DLS, TEM and XRD methods. The Dynamic Light Scattering (DLS) showed the particle size distribution of the AgNPs synthesized from C. roseus var. alba callus extract was found 86.95 nm with polydispersity index (PDI) value of = 0.304. TEM images showed the formation of AgNPs with an average size of 10 nm to 20 nm. And the XRD analysis showed that the AgNPs were crystalline in nature with face-centered-cubic (FCC). For the assessment of antibacterial activity the concentration of AgNPs 25 μL, 50 μL, 100 μL and 150 μL were used against both the bacterial strain S. aureus and P. aeruginosa, the zone of inhibition found 4 mm, 7 mm, 16 mm and 23 mm as well as 5 mm, 9 mm, 19 mm and 26 mm respectively.
Conclusions: Aseptically engendered callus of C. roseus var. alba demonstrates vigorous potential for synthesis of silver nanoparticles by rapid reduction of Ag+ to Ag. The biologically synthesized AgNPs showed more preponderant antibacterial effect against S. aureus and P. aeruginosa.

Open Access Original Research Article

Adherence of Irradiated Slime Producing Bacterial Pathogens to Biomaterial Surface and their Antimicrobial Susceptibility Associated with Catheter Infection in Bladder Cancer Patients

Hala Abd Allah Farrag, Al Zahraa A-Karam El-Din, Zeinab Galal Mohamed El-Sayed, Mona Mohamed Kamal

Journal of Pharmaceutical Research International, Page 1604-1628
DOI: 10.9734/BJPR/2014/9908

Aims: To detect the prevalence of biofilm producers among Gram negative bacilli and Gram positive cocci bacterial pathogens along with their antimicrobial susceptibility pattern. Growth and adherence on catheter eluates and in the presence of antibiotics.
Methodology: From laboratory of microbiology, one hundred samples (100 urinary catheters and 100 urine samples from the attached drainage bags) of bladder cancer patients collected in National Cancer Institute in Cairo, Egypt, were identified to species level. Slime production was investigated by the quantitative and qualitative methods. Qualitative method was carried out by tube method. Adherence assay and quantitation of biofilm was performed by spectrophotometric method by measuring the optical densities of stained bacterial films adherent to plastic tissue culture plates. Hydrophobicity was evaluated by adhesion to P-xylene. Identification and minimum inhibitory concentration (MICs) of 26 antimicrobial agents against gram negative and 24 against gram positive bacterial isolates were determined using microscan walk away 96 SI system. Plasmid profile analysis was carried out by plasmid isolation kit. Scanning electron microscopy studies for growth, adherence and biofilm formation. Impact of gamma irradiation at a dose level of 24.41Gy was studied.
Results: From the processing of 100 samples, 98 cases were positive. Out of them 110 isolates of gram negative bacilli and 13 of gram positive cocci. They were belonging to 15 and 6 species respectively. Among them, 117 isolates showed positive results for adherence assay and biofilm/slime production. They were identified as; Escherichia coli, Klebsiella spp., Enterobacter spp., Acinetobacter, Proteus spp., Citrobacter, Alcaligenes, Empedobacter (104 strains) Staphylococcus spp. and Enterococcus (13 stains). The results obtained by different methods correlated well with strain to strain variation. Gamma irradiation resulted in changes in slime production and adherence ability for all the tested strains. Cell surface hydrophobicity (CSH) showed a hydrophobic reaction and these with increase in its value after irradiation in case of Escherichia coli. On the other hand, Staphylococcus epidermidis was moderate hydrophobic before irradiation changed to strictly hydrophilic after irradiation. All the slime producers showed reduced susceptibility to majority of antibiotics. They exhibited highest percentage susceptibility before and after in vitro gamma irradiation at a dose level 24.41Gy for both Amikacin and Imipenem. Scanning electron microscopy (SEM) confirmed growth and biofilm formation in the presence of catheter eluates only with halos surrounding the cells and visible erosion zones on catheter surfaces. The antimicrobial and adherence activity of Amikacin and Imipenem at the MIC level showed marked abnormalities in cells shape and size with significant reduction in adherence ability. Plasmid profile analysis of irradiated strains showed more extra-plasmid bands and / or difference in molecular weight.
Conclusion: The biofilm assay strategy applied in this study may constitute a tool in biomaterial related infection and antimicrobial resistant research for further studies for biomaterial modification. Early detection of biofilm forming organisms can help in appropriate antibiotic choice.

Open Access Original Research Article

Simultaneous Estimation of Luteolin and Apigenin in Methanol Leaf Extract of Bacopa monnieri Linn by HPLC

Aiyalu Rajasekaran, Nagulsamy Sarathikumar, Vivekanandan Kalaiselvan, Muthusamy Kalaivani

Journal of Pharmaceutical Research International, Page 1629-1637
DOI: 10.9734/BJPR/2014/9009

Aims: To develop, validate and quantify the content of flavonoids luteolin and apigenin in aerial parts of methanol leaf extract of Bacopa monnieri (B. monnieri) by reverse phase liquid chromatography.
Study Design: High Performance Liquid Chromatography
Place and Duration of Study: Department of Pharmaceutical Analysis, KMCH College of Pharmacy, Coimbatore and Indian Pharmacopoeia Commission, Ghaziabad, India from 1.7.2010 to 30.6.2011.
Methodology: Separation and quantification of flavonoids was performed on a C18 column (5µm, 200mm×4.6mm, id.) using potassium dihydrogen phosphate buffer (20mM, pH 3.5 adjusted with ortho phosphoric acid, v/v) and acetonitrile as mobile phase with a flow rate of 1ml/min. The column effluents were monitored at 348nm with column temperature kept at 30±1ºC.
Results: A validated method for simultaneous estimation of luteolin and apigenin was developed, where limits of detection and quantification was found to be 0.03 and 0.91µg/ml for luteolin, 0.041 and 0.13µg/ml for apigenin respectively. The percentage of these phytoconstituents recovered was in the range of 98.07-99.71 (%RSD<2%).
Conclusion: The developed validated HPLC method was found to be accurate, precise and robust and may be used for analysis of luteolin and apigenin in the extracts of B. monnieri.

Open Access Original Research Article

Evaluation of the Anxiolytic Effect of Methanolic Leaves Extract of Paullinia pinnata Lin in Mice

M. Aliyu, J. A. Anuka, A. H. Yaro, M. G. Magaji

Journal of Pharmaceutical Research International, Page 1638-1646
DOI: 10.9734/BJPR/2014/10990

This study was designed to evaluate the anxiolytic effect of methanolic leave extract of Paullinia pinnata L. in mice. The elevated plus-maze and staircase paradigm were used to assess the anxiolytic activity of the methanolic leaf extract of Paullinia pinnata and diazepam. The results of the elevated plus-maze test showed that the extract at the dose of 50 mg/kg and diazepam significantly (P<0.05 and P<0.005) increased both the number of entries and time spent in the open arm by mice. In staircase paradigm, the extract produced a significant (P<0.05-P<0.0005) dose dependent decrease in the number of steps ascended and number of rearing events compared to the control mice. Diazepam significantly (P<0.0005) reduced the number of rearing events compared to control. The result of the present preliminary study suggests that methanolic leaf extract of Paullinia pinnata may possess an anxiolytic activity.

Open Access Original Research Article

Drying Kinetics of Gum Karaya (Sterculia urens) Using Convective Air Dryer

Jaspreet Singh, Balwinder Singh

Journal of Pharmaceutical Research International, Page 1647-1656
DOI: 10.9734/BJPR/2014/10104

The present study was aimed to investigate the drying kinetics of gum karaya (S. urens). Three grades of gum (I, II, III) were dried using a cabinet-type convective dryer. Particle size equivalent to US Sieve size-6 and air temperatures of 50, 60 and 70°C were used for the drying experiments. The experimental drying data was fitted to Page’s model to predict the drying kinetics. Investigations with constant air velocity revealed that for grade-I gum, drying rate constant (k) varied between 0.2744-0.3742 (h-1), for grade-II gum between 0.3208-0.4439 (h-1) and for grade-III gum between 0.4098-0.4639 (h-1). The dimensionless number (n) was always more than 1 and minimum value of the Coefficient of determination (R2) was 0.967. Increase in air temperature enhanced drying process and drying rate. For each particular temperature, the values of drying rate constant were minimum for grade-I, maximum for grade-III and intermediate for grade-II gum.

Open Access Original Research Article

Quality Assessment of Some Brands of Tetanus Toxoids Marketed in Open Markets in South-Eastern Nigeria

V. U. Chigozie, P. M. Eze, C. R. Chukwunwejim, C. C. Abba, C. S. Nworu, C. O. Esimone

Journal of Pharmaceutical Research International, Page 1657-1667
DOI: 10.9734/BJPR/2014/10901

Aims: This study was designed to verify the immunogenicity (potency) and the safety of tetanus toxoid vaccines marketed in three large open drug markets in South-Eastern Nigeria.
Methodology: Tests for Sterility, formaldehyde concentrations, specific toxicity, endotoxin, and immunogenicity (potency) were conducted on three different brands of tetanus toxoids (Brand α - from Ariaria Drug Market, Aba-Abia State; Brand β - from Ogbete Drug Market, Enugu State; and Brand γ - from Bridge-Head Drug Market, Onitsha-Anambra State).
Results: All vaccine brands studied passed the sterility testing, but did not comply with the 2011 BP specifications on free formaldehyde concentration, which stipulates that the free formaldehyde concentration should not exceed 0.02%. The three vaccine brands did not show specific, abnormal, or general toxicity, but contained different amounts of endotoxins. The result of the potency testing showed that the three brands were immunogenic and elicited specific antibodies against tetanus toxin; but brand γ was the most immunogenic since it elicited the highest titers of total IgG, IgG1, and IgG2a followed by brand α, and then brand β. 
Conclusion: Generally, the quality control tests carried out on these three commercial brands of tetanus toxoids marketed in Nigeria showed that they do not comply with all the pharmacopeial standards on quality and safety required for vaccines of this nature. Therefore, we conclude that some of the tetanus toxoids marketed in open drug markets in Nigeria are substandard and may be responsible for the failures of these vaccines used for immunization in the country.

Open Access Original Research Article

An Assessment of the Level of Awareness, Attitudes, Opinions of Pharmacy Students Concerning HIV and AIDS in Malaysia

Rohit Kumar Verma, Shradha Batuk Bavisi, Syed Imran Ahmed, Ankur Barua

Journal of Pharmaceutical Research International, Page 1668-1676
DOI: 10.9734/BJPR/2014/7455

Aims: HIV and AIDS spreading wide and causing serious threats and deaths among Malaysian residents. A nationwide cross-sectional survey was conducted to assess the awareness, attitudes and opinions about HIV and AIDS among pharmacy students.
Methods: A total of 316 pharmacy students of year three and onwards took part in the survey. Students were asked to fill in questionnaires with consent forms. The results were analyzed by using SPSS version 17.
Results: The data indicated that awareness about HIV and AIDS was moderate. High level of awareness was seen for major routes of HIV transmissions, but lower level of awareness was seen for other modes of transmission like circumcision, visiting barbers, and blood splashes on outer body surface. Only 19.3% and 13.3% of respondents were aware about HIV prevention by sex abstinence and by staying faithful to one partner respectively. The respondents had doubts in keeping HIV and AIDS patients in close vicinity to them and their family.
Conclusion: According to the findings, the respondents had a few misconceptions about HIV transmission and prevention. Data from this survey may be useful to hold programs and campaigns designed to convey accurate information about HIV transmission and prevention. Talks and media campaigns should also be carried out to change their attitudes and opinions about HIV and AIDS.

Open Access Original Research Article

Pharmacological Activities of Blumea lacera (Burm. f) DC: A Medicinal Plant of Bangladesh

Abul Khair, Mohammed Ibrahim, Qamrul Ahsan, Zilly Homa, Md. Ruhul Kuddus, Ridwan Bin Rashid, Mohammad Abdur Rashid

Journal of Pharmaceutical Research International, Page 1677-1687
DOI: 10.9734/BJPR/2014/10001

Aims: The crude methanol extract of whole plant of Blumea lacera (Burn.f.) DC. has been investigated for anti-diarrheal, antimicrobial, anxiolytic, anti-atherothrombosis, membrane stabilizing and alpha-amylase inhibitory activities.
Place and Duration of Study: The study was carried out in 2013 in the Department of Pharmacy, Southern University Bangladesh, Chittagong, Bangladesh.
Methodology: Test for anti-diarrheal activity was carried out by castor oil-induced diarrhea in mice. The preliminary antimicrobial activity was determined by the agar disc diffusion method. The anxiolytic activity was examined in mice by using the hole board test and open field test (OFT). The anti-atherothrombosis activity was evaluated using standard streptokinase. The membrane stabilizing activity was assessed by using hypotonic solution induced hemolysis of human erythrocyte. The plant extract was also assessed for anti-diabetic ability using In vitro α-amylase inhibitory potential. The α-amylase inhibitory activity of B. lacera was measured using the starch-iodine method.
Results: The crude extract of B. lacera showed anti-diarrheal activity in dose-dependent manner. In antimicrobial assay, this extract showed better activity against the tested fungi compared to the bacteria used in the screening. Significant anxiolytic activity was found for this plant extract. In the In vitro anti-atherothrombosis test, the extract exhibited 46.17% clot lysis as compared to the standard, streptokinase (81.53%). In membrane stabilizing activity test, the plant extract at 1.0mg/ml inhibited the heat-induced hemolysis of RBCs by 52.27% whereas the standard acetyl salicylic acid (ASA) demonstrated 81.72% inhibition of hemolysis. Our results revealed that the extract had dose dependent prevention of digestion of carbohydrates by inhibiting α-amylase. The ability of B. lacera to inhibit thermal-and hypotonic-enzyme activity was found to be statistically significant (p=0.05).
Conclusion: These results demonstrated that B. lacera may be used in pharmaceutical applications because of its effective pharmacological properties.

Open Access Original Research Article

Comparative Quality Control Evaluation of Atenolol Tablets Marketed in Kuala Lumpur, Malaysia

Senthil Rajan Dharmalingam, Muhammad Azizi, Suresh Shanmugham, Venkata Srikanth Meka, Wong Pei Se

Journal of Pharmaceutical Research International, Page 1688-1695
DOI: 10.9734/BJPR/2014/8813

Background: The emergent of many pharmaceutical companies producing their own generic type of drugs after the patent of innovator drugs expired can improve the general healthcare delivery systems as well as decreasing the healthcare costs. But it also raises a few issues with one of it is the widespread of substandard and counterfeit product. Post-surveillance study to assess product parameter of various generics drug marketed is crucial. This kind of monitoring reduces a country’s economical burden on health issues from diseases due to fraudulent and substandard drugs usage.
Purpose: The main objective of this study is to perform a comparative evaluation of the physicochemical properties of five commercially available leading brands of Atenolol tablets marketed in Kuala Lumpur.
Method: The quality control parameters of five different brands of atenolol tablets were atenolol tablet assessed included uniformity of content, uniformity of weight, friability, crushing strength, disintegration and dissolution tests as well as content uniformity of the tablets. All the tablets were assessed for conformity with British Pharmacopoeia (BP) standards.
Results: All the five brands of the tablets passed the British Pharmacopoeia (BP) standards for weight uniformity, disintegration, friability, content uniformity and hardness tests.
Conclusion: The quality control parameters of all five top selling brands of atenolol tablets marketed in Kuala Lumpur analyzed passed all the BP and USP quality specifications and were physically and chemically equivalent.