Advanced Pharmaceutical Bulletin
Volume:2 Issue: 2, Dec 2012

The aim of this study was to develop a simple, rapid and sensitive reverse phase high performance liquid chromatography (RP-HPLC) method for quantification of sirolimus (SRL) in pharmaceutical dosage forms. The chromatographic system employs isocratic elution using a Knauer- C18, 5 mm, 4.6 × 150 mm. Mobile phase consisting of acetonitril and ammonium acetate buffer set at flow rate 1.5 ml/min. The analyte was detected and quantified at 278nm using ultraviolet detector. The method was validated as per ICH guidelines. The standard curve was found to have a linear relationship (r2 > 0.99) over the analytical range of 125–2000ng/ml. For all quality control (QC) standards in intraday and interday assay, accuracy and precision range were -0.96 to 6.30 and 0.86 to 13.74 respectively, demonstrating the precision and accuracy over the analytical range. Samples were stable during preparation and analysis procedure. Therefore the rapid and sensitive developed method can be used for the routine analysis of sirolimus such as dissolution and stability assays of pre- and post-marketed dosage forms.

The aim of the present research was to formulate a transparent microemolsion as a topical delivery system for tretinoin for the treatment of acne. Microemulsion formulations prepared by mixing appropriate amount of surfactant including Tween 80 and Labrasol, co-surfactant such as propylene glycol (PG) and oil phase including isopropyl myristate – transcutol P (10:1 ratio). The prepared microemolsions were evaluated regarding their particle size, zeta potential, conductivity, stability, viscosity, differential scanning calorimetry (DSC), scanning electron microscopy (SEM), refractory index (RI) and pH. The results showed that maximum oil was incorporated in microemolsion system that was contained surfactant to co-surfactant ratio (Km) of 4:1. The mean droplets size range of microemulsion formulation were in the range of 14.1 to 36.5 nm and its refractory index (RI) and pH were 1.46 and 6.1, respectively. Viscosity range was 200-350 cps. Drug release profile showed 49% of the drug released in the first 8 hours of experiment belong to ME-7. Also, Hexagonal and cubic structures were seen in the SEM photograph of the microemulsions. physicochemical properties and in vitro release were dependent upon the contents of S/C, water and, oil percentage in formulations.Also, ME-7 may be preferable for topical tretinoin formulation.

Long term L-DOPA therapy in Parkinson’s disease is associated with troublesome motor fluctuations such as L -DOPA Induced dyskinesia and wearing off effect. Our recent study showed that activation of 5-HT1A receptors could improve the anti-cataleptic effect of L-DOPA in parkinsonian rats. In this study we investigated the effect of fluoxetine on anti-parkinsonian effect of L-DOPA in 6-hydroxydopamine (6-OHDA)-lesioned rats. Catalepsy and motor incoordination were induced by unilateral injection of 6-OHDA (8µg/2µl/rat) into the central region of the sabstantia nigra pars compacta (SNc). After 3 weeks as a recovery period, these rats injected intraperitoneally (i.p.) L-DOPA (15 mg/kg) twice daily for 20 consecutive days, and anti-parkinsonian effect of L-DOPA was investigated by bar-test and rotarod on days 5, 10, 15 and 20. The results showed that L-DOPA is able to improve motor coordination in rotarod only until day 15 and these effects of L-DOPA were abolished on the day 20. On day 21, rats were co-injected with fluoxetine (0.1, 0.5 and 1mg/kg, i.p.) and L-DOPA (15 mg/kg, i.p.). Fluoxetine increased anti-cataleptic effect of L-DOPA at the dose of 1 mg/kg, while fluoxetine had not any impact on the effect of L-DOPA in rotarod test. The effect of fluoxetine (1 mg/kg, i.p.) on anti-cataleptic effect of L-DOPA (15 mg/kg, i.p.) was reversed by 1-(2-methoxyphenyl)-4-(4-phthalimidobutyl) piperazine hydrobromide (NAN-190; 0.5 mg/kg, i.p.), as a 5-HT1A receptor antagonist. According to the results, it may be concluded that fluoxetine improves 6-OHDA-induced catalepsy and motor imbalance in L-DOPA- treated rats through activation of 5-HT1A. Further studies should be designed to clarify the precise mechanism of interaction between 5-HT1A and dopaminergic neurons.

Fluoxetine is the most prescribed antidepressant drug worldwide. In this work, a new dispersive liquid–liquid microextraction (DLLME) method combined with spectrofluorimetry has been developed for the extraction and determination of FLX in pharmaceutical formulations and human urine. For FLX determination, the pH of a 10 mL of sample solution containing FLX, was adjusted to 11.0. Then, 800 µL of ethanol containing 100 µL of chloroform was injected rapidly into the sample solution. A cloudy solution was formed and FLX extracted into the fine droplets of chloroform. After centrifugation, the extraction solvent was sedimented and supernatant aqueous phase was readily decanted. The remained organic phase was diluted with ethanol and its fluorescence was measured at 292±3 nm after excitation at 234±3 nm. Some important parameters influencing microextraction efficiency were investigated. Under the optimum extraction conditions, a linear calibration curve in the range of 10 to 800 ng/mL with a correlation coefficient of r2 = 0.9993 was obtained. Limit of detection (LOD) and limit of quantification (LOQ) were found to be 2.78 and 9.28 ng/mL, respectively. The relative standard deviations (RSDs) were less than 4%. Average recoveries for spiked samples were 93–104%. The proposed method gives a very rapid, simple, sensitive, wide dynamic range and low–cost procedure for the determination of FLX.

In the present study, effects of preischemic administration of natural honey on cardiac arrhythmias and myocardial infarction size during zero flow global ischemia were investigated in isolated rat heart. The isolated hearts were subjected to 30 min zero flow global ischemia followed by 120 min reperfusion then perfused by a modified drug free Krebs-Henseleit solution throughout the experiment (control) or the solution containing 0.25, 0.5, 1 and 2% of natural honey for 15 min before induction of global ischemia (treated groups), respectively. Cardiac arrhythmias were determined based on the Lambeth conventions and the infarct size was measured by computerized planimetry. Myocardial infarction size was 55.8±7.8% in the control group, while preischemic perfusion of honey (0.25, 0.5, 1 and 2%) reduced it to 39.3±11, 30.6±5.5 (P<0.01), 17.9±5.6 (P<0.001) and 8.7±1.1% (P<0.001), respectively. A direct linear correlation between honey concentrations and infarction size reduction was observed (R2=0.9948). In addition, total number of ventricular ectopic beats were significantly decreased by all used concentrations of honey (P<0.05) during reperfusion time. Honey (0.25, 0.5 and 1 %) also lowered incidence of irreversible ventricular fibrillation (P<0.05). Moreover, number and duration of ventricular tachycardia were reduced in all honey treated groups. Preischemic administration of natural honey before zero flow global ischemia can protect isolated rat heart against ischemia/reperfusion injuries as reduction of infarction size and arrhythmias. Maybe, antioxidant and free radical scavenging activities of honey, reduction of necrotized tissue and providing energy sources may involve in these cardioprotective effects of honey.

The present study was designed to explore the effect of intraperitoneal administration of cisplatin in germinal epithelium of mice. There are few reports on the side effect of cisplatin on spermatogenesis when are used as anticancer drug. Therefore, in the present study the effect of cisplatin on spermatogenesis was evaluated by electron microscopy. Twenty balb/c mice aging 6-8 weeks was used in this study. The mice were divided into two groups, control and cysplatin treated. cysplatin was injected for five days as 2.5 mg /kg. The mice were sacrificed after 5 weeks and testicular specimens were removed, fixed in boueins, formaldeyd fixative and 2.5% Glutaraldehide then prepared for light and electron microscopic study. Observation with optic microscope in treated group thickness of germinal epithelium was reduced a lot and increased the number of apoptotic cells. In some seminiferous tubules only sertoli cells were observed and nucleus of spermatogony cells was hetrochromatin. The electron microscopic observations showed some irregularity waviness and thickening in basal layer. Also myoid cells of this group were thick and contracted. In this group many apoptotic cells and damaged organelles were seen. It was indicated that cisplatin affected testicular germinal epithelium by both cytotoxic effect and induction of apoptosis.

The objective of the present study was to evaluate the protective effect of methanol extract of Prosopis cineraria (MPC) against N-nitrosodiethylamine (DEN, 200mg/kg) induced Phenobarbital promoted experimental liver tumors in male Wistar rats. The rats were divided into four groups, each group consisting of six animals. Group 1 served as control animals. Liver tumor was induced in group 2, 3, and 4 and Group 3 animals received MPC 200mg/kg and Group 4 animals received MPC 400mg/kg. Administration of DEN has brought down the levels of membrane bound enzymes like Na+/ K+ ATPase, Mg2+ ATPase and Ca2+ATPase which were later found to be increased by the administration of Prosopis cineraria (200 and 400mg/kg) in dose dependent manner. The MPC extract also suppressed the levels of glycoproteins like Hexose, Hexosamine and Sialic acid when compared to liver tumor bearing animals. Our study suggests that MPC may extend its protective role by modulating the levels of membrane bound enzymes and suppressing glycoprotein levels.

Dry powder inhaler (DPI) is generally formulated as a powder mixture of coarse carrier particles and micronized drug with aerodynamic diameters of 1–5 μm. Carrier particles are used to improve drug particle flowability, thus improving dosing accuracy, minimizing the dose variability compared with drug alone and making them easier to handle during manufacturing operations. Lactose is the most common and frequently used carrier in DPIs formulations and nowadays various inhalation grades of lactose with different physico-chemical properties are available on the market. Therefore, the purpose of this manuscript is to review evolution of lactose as a carrier in inhalable formulations, their production and the impact of its physico-chemical properties on drug dispersion. This review offers a perspective on the current reported studies to modify lactose for better performance in DPIs.

Results of our previous study revealed that preischemic perfusion of honey before zero flow global ischemia had cardioprotective effects in rat. The present study investigated potential resistance to reperfusion injury following short term postischemic administration of natural honey in globally ischemic isolated rat heart. Male Wistar rats were divided into five groups (n=10-13). The rat hearts were isolated, mounted on a Langendorff apparatus, allowed to equilibrate for 30 min then subjected to 30 min global ischemia. In the control group, the hearts were reperfused with drug free normal Krebs-Henseleit (K/H) solution before ischemia and during 120 min reperfusion. In the treatment groups, reperfusion was initiated with K/H solution containing different concentration of honey (0.25, 0.5, 1 and 2%) for 15 min and was resumed until the end of 120 min with normal K/H solution. In the control group, VEBs number was 784±199, while in honey concentration of 0.25, 0.5, 1 and 2%, it decreased to 83±23 (P<0.001), 138±48 (P<0.01), 142±37 (P<0.001) and 157±40 (P<0.01), respectively. Number and duration of VT and time spent in reversible VF were also reduced by honey. In the control group, the infarct size was 54.1±7.8%, however; honey (0.25, 0.5, 1 and 2%) markedly lowered the value to 12.4±2.4, 12.7±3.3, 11.3±2.6 and 7.9±1.7 (P<0.001), respectively. Postischemic administration of natural honey in global ischemia showed protective effects against ischemia/reperfusion (I/R) injuries in isolated rat heart. Antioxidant and radical scavenging activity, lipoperoxidation inhibition, reduction of necrotized tissue, presence of rich energy sources, various type of vitamins, minerals and enzymes and formation of NO-contain metabolites may probably involve in those cardioprotective effects.

Ginger, the rhizome of Zingiber officinale, is one of the most widely used spices for various foods and as an herbal medicine in Asian countries. It has been shown that ginger has antioxidant power. Gentamicin is an aminoglycoside antibiotic with a very broad spectrum against microbial pathogens, especially the gram-negative. Many studies revealed that gentamicin induces an oxidative stress-status in the testis by increasing free radical formation and lipid peroxidation. The present study was designed to investigate on the effects of Ginger as a natural anioxidant on testis apoptosis after treatment with gentamicin in rats. In order to study the recovery effects of ginger on testis apoptosis after treatment with gentamicin 40 adult Wistar male rats were selected and randomly divided into four groups. Normal salin control (group I) (n=10), gentamicin control (group II), ginger control (group III) and gentamicin + ginger (group IV) each 10 rats. There was observation of negative effect of Gentamicin on testis histology in rats. The results revealed that there was a significant increase in apoptosis in group III when compared with other groups (P<0.05).However, ginger could decrease apoptosis in group IV that received 100mg/kg/rat of Ginger. Regarding the results, it is recommended that administration of ginger with gentamicin might be beneficial in men who receive gentamicin to treat infections.

Atrial fibrillation (AF) is one of the most common types of sustained dysrhythmia and there are some disagreements about its treatment. The goals of AF treatment include the control of ventricular rate, the establishment of sinus rhythm and the prevention of thromboembolic events. In this study, the effect of verapamil was compared to digoxin on heart rate control in patients with AF. This descriptive study was conducted in an emergency department (ED) in Iran. Sixty patients with a new onset AF and rapid ventricular response receiving digoxin or verapamil were included and observed. Two thirty-patient groups receiving verapamil or digoxin were evaluated. The heart rate was significantly decreased in both groups (p = 0.002); however, the cardioversion was not noticed in both of them. The best rate control in verapamil and digoxin groups was observed after ​​5.9 mg (46.7%) and 0.6 mg (36.7%), respectively. Administration of verapamil in comparison with digoxin has no difference to control the heart rate in AF patients. It should be taken into consideration that prospective randomized studies should be conducted to identify the efficacy and select the best of these two drugs to treat AF patients.

In this study, three important medicinal plants (Enhydra fluctuans Lour, Clerodendrum viscosum Vent and Andrographis peniculata Wall) of Bangladesh were investigated to analyze their antimicrobial and cytotoxic activities against some pathogenic microorganisms and Artemia salina (brine shrimp nauplii). The coarse powder material of leaves of each plant was extracted separately with methanol and acetone to yield methanol extracts of leaves of Enhydra fluctuans (MLE), Clerodendrum viscosum (MLC) and Andrographis peniculata (MLA), and acetone extracts of leaves of Enhydra fluctuans (ALE), Clerodendrum viscosum (ALC) and Andrographis peniculata (ALA). The disc diffusion method and the method described by Meyer were used to determine the antimicrobial and cytotoxic activities of each plant extract. Among the test samples, MLE and ALE showed comparatively better antimicrobial activity against a number of bacteria and fungi with inhibition zones in the range of 06-15 mm and according to the intensity of activity, the efficacy against microorganisms were found in the order of Enhydra fluctuans > Andrographis peniculata > Clerodendrum viscosum. In cytotoxicity assay, all samples were found to be active against brine shrimp nauplii (Artemia salina) and ALA produced lowest LC50 value (7.03 μg/ml). Enhydra fluctuans and Andrographis peniculata possesses significant antimicrobial and cytotoxic activities.

An efficient method has been described for synthesis of 6-(substituted aryl)-4-(3, 5-diphenyl-1H-1,2,4-triazol-1-yl)-1, 6-dihydropyrimidine-2-thiol, as a beneficial antimicrobial, anticonvulsant and anticancer agents. The clalcones of title compounds were synthesized in three steps and subsequently these chalcones were further reacted with thiourea in the presence of KOH in ethanol, which led to the formation of dihydropyrimidine derivatives (4a-j). Compounds 4a-j were screened for their in vitro antimicrobial activity by agar well method and their anticonvulsant activity by the MES model. Anticancer activity of two newly synthesized heterocycles were evaluated at National Cancer Institute (NCI) Maryland, USA against 60 cell lines of different human tumor at a single dose of 10-5M. Compound 4b, 4c, 4d, 4i and 4j were exhibited significant antimicrobial potential against tested strains at 50μg/ml and 100μg/ml concentrations. Out of the ten compounds studied 4a, 4b, 4c, 4h and 4j showed comparable MES activity to Phenytoin and Carbamazepine after 0.5h. Tested compounds did not showed to be more potent than standard drugs after 4h. Compound 4a and 4d were found active on Non-Small Cell Lung Cancer (HOP-92) Ten novel dihydropyrimidine analogues has been synthesized, characterized and found to be promising antibacterial, anticonvulsant and antitumor agents.

Breast cancer is the most common cause of cancer-related death in women world wide. Therefore, there is an urgent need to identify and develop therapeutic strategies against this deadly disease. This study is the first to investigate the cytotoxic effects and the mechanism of cell death of Scrophularia oxysepala extracts in MCF-7 human breast cancer cells. Three extracts of Scrophularia oxysepala including the n-hexane, dichloromethane and methanol extracts were examined. MTT (3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and Trypan-blue assays were performed in MCF-7 cells as well as Human umbilical vein endothelial cells (HUVEC) to analyze the cytotoxic activity of the extracts of Scrophularia oxysepala. Further, the apoptosis inducing action of the extracts was determined by TUNEL (terminal deoxy transferase (TdT)-mediated dUTP nick- end labeling) test and cell death assay. The results showed that the n-hexane extract had no cytotoxic effects but dichloromethane and methanol extracts significantly inhibited cell growth and viability in a dose and time dependent manner without inducing damage to non-cancerous cell line HUVEC. In addition, Cell death assay and DNA fragmentation analysis using TUNEL indicated induction of apoptosis by dichloromethane and methanol extracts of Scrophularia oxysepala in MCF-7 cells. Our studies suggest that this plant may contain potential bioactive compound(s) for the treatment of breast cancer.

This study was performed to investigate the incidence of thrombophilic gene mutations in repeated assisted reproductive technology (ART) failures. The prevalence of mutated genes in the patients with a history of three or more previous ART failures was compared with the patients with a history of successful pregnancy following ARTs. The study group included 70 patients, 34 with three or more previously failed ARTs (A) and control group consisted of 36 patients with successful pregnancy following ARTs (B). All patients were tested for the presence of mutated thrombophilic genes including factor V Leiden (FVL), Methylenetetrahydrofolate reductase (MTHFR) and Prothrombin (G20210A) using real-time polymerase chain reaction (RT- PCR). Mutation of FVL gene was detected in 5.9% women of group A (2 of 34) compared with 2.8% women (1 of 36) of control group (P = 0.6). Mutation of MTHFR gene was found in 35.3% (12 cases) as compared with 50% (18 cases) of control (35.3% versus 50%; P = 0.23). Regarding Prothrombin, only control group had 5.6% mutation (P = 0.49). No significant differences were detected in the incidences of FVL, Prothrombin and MTHFR in the study group A compared with the control group B. The obtained results suggest that thrombophilia does not have a significant effect in ART failures.

The purpose of the present work was to assess iontophoretic permeation of Lisinopril at different current densities and concentrations for development of patient-controlled active transdermal system. In vitro iontophoretic transdermal delivery of Lisinopril across the pigskin was investigated at three different drug concentrations and three different current densities (0.25- 0.75 mA/cm2) in the donor cell of the diffusion apparatus, using cathodal iontophoresis along with the passive controls. For passive permeation, the steady state flux significantly increased with the increasing of donor drug concentration. At all concentration levels, iontophoresis considerably increased the permeation rate compared to passive controls. Iontophoretic transport of Lisinopril was to be found increase with current densities. Flux enhancement was highest at the lowest drug load and lowest at the highest drug load. The obtained results indicate that permeation rate of Lisinopril across the pigskin can be considerably enhanced, controlled or optimized by the use of Iontophoresis technique.

The increasing use of herbal drugs and their easy availability have necessitated the use of mutagenicity test to analyze their toxicity and safety. The aim of this study was to evaluate the genotoxicity of Supermint herbal medicine in DNA breakage of rat hepatocytes in comparison with sodium dichromate by single cell gel electrophoresis technique or comet assay. Hepatocytes were prepared from male wistar rats and were counted and kept in a bioreactor for 30 minutes. Then cells were exposed to the Supermint herbal medicine at doses of 125, 250 and 500 µl/ml. Buffer 4 (incubation buffer) and sodium dichromate were used as negative and positive control for one hour respectively. Then cell suspension with low melting point agarose were put on precoated slides and covered with agarose gel. Then lysing, electrophoresis, neutralization and staining were carried out. Finally the slides were analyzed with fluorescence microscope. The parameter under this analysis was the type of migration which was determined according to Kobayashi pattern. With increased dose of Supermint herbal medicine the DNA damage was slightly increased (P<0001). Conlusion: In overall compared to the positive control significant differences is observed which convinced that the crude extract of Supermint in vitro did not have mutagenic effect.

Patients with increased intracranial pressure (ICP) are prone to severe cardiac and or cerebral complications following emergence from general anesthesia and especially post-extubation phase. Administering beta blockers including esmolol is believed to be helpful in providing a stable hemodynamic at the end of the surgery and recovery stages and reducing recovery phase length. In a double-blind prospective randomized clinical trial, 60 adult patients with ASA (American Society of Anesthesiologist) class of I-II scheduled to undergo elective neurosurgery operations were randomly divided into two groups receiving esmolol (n=30) and placebo (n=30) as IV infusion within four minutes prior to extubation continued by an IV infusion for 10 minutes after extubation. There was a significant difference between two groups regarding the changes of systolic blood pressure and heart rate at all studied stages after extubation (P≤0.05). However, no significant difference existed between esmolol and control groups regarding recovery and extubation times emphasizing the fact that esmolol is of excellent early recovery and extubation profiles. Esmolol is advised to be used in preventing hyperdynamic status throughout extubation phase without extending recovery phase length.

Direct tableting has been renewed as a preferable process by simply mixing and compressing powder to save time and cost in comparison with granule tableting. Direct compression tableting as a technique has been successfully applied to numerous drugs on the industrial scale, although the success of any procedure, and resulting mechanical properties of tablets, is strongly affected by the quality of the crystals used. Good flowability, packability and compactability are prerequisite for drug to be prepared by direct tableting. When the mechanical properties of the drug particles are inadequate a primary granulation is necessary. The use of spherical crystallization as a technique appears to be an efficient alternative for obtaining suitable particles for direct tableting. Spherical crystallization is a particle design technique, by which crystallization and agglomeration can be carried out simultaneously in one step and which has been successfully utilized for improvement the micromeritic properties of crystalline drugs. In this review, we will discuss how the micromeritic properties of the particles such as flowability, packability and compactability can be improved by spherical crystallization technique.

This study is aimed at determining chemical constituents and antimicrobial activities of a common brand of black tea (Lipton®) in Nigeria. Standard methods were employed for testing carbohydrates, tannins, saponins, flavonoids, alkaloids, steroids and terpenes in the tea. Antimicrobial activities of methanolic and aqueous extracts of the tea on four standard strains of organisms: Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis were also determined by standard methods. Results showed that the tea contains tannin and reducing sugar. Concentrations of 1%, 2%, 4%, 6%, 8% and 10% aqueous and methanolic extract of black tea were prepared and their zones of inhibition determined against the four test organisms using the cup plate method. This was compared with zones for standard disc Gentamicin (10 µg) and Erythromycin (15 µg). Pseudomonas aeruginosa was sensitive to 2% to 10% aqueous extracts and intermediate to 6%, 8% and 10% methanolic extracts. E. coli was intermediately sensitive to 6%, 8% and 10% aqueous extract and 2% to 10% methanolic extracts. B. subtilis was intermediately sensitive to 4%, 6% and 8% aqueous extract and 4% to 10% methanolic extract but sensitive to 10% aqueous extract. Staph.aureus was intermediately sensitive to 4% to 10% aqueous extracts and 2% to 10% methanolic extracts. B. subtilis had the lowest MIC values of both aqueous and methanolic extracts. In conclusion, this study has shown that Lipton® has antimicrobial properties on E.coli, Staph.aureus, B.subtilis and Ps.aeruginosa and contains tannin and reducing sugar.