Pharmaceutical Sciences
Volume:22 Issue: 2, Jun 2016

Hesperidin is a flavonoid and has strong anti-oxidant and anti-inflammatory activities. The aim of the present study is to investigate the effects of hesperidin on ischemic/reperfusion (I/R) induced injuries and arrhythmias.
Male Wistar rats were anesthetized and then the hearts were removed and cannulated immediately to a langendorff apparatus and prepared for the left coronary artery ligation. The hearts were perfused with Krebs-Henseleit Buffer Solution (KHBS; control) or KHBS plus hesperidin (1, 2.5 & 5µg/ml; treated groups) 5 minutes before coronary occlusion, during the ischemia, and reperfusion period. After reperfusion, double staining strategy (Evans blue and TTC) were used. The percentage of infarct size was determined by Image-j software. Arrhythmia in control group and treated groups were analyzed and compared. Lactate concentration was measured in samples at the end of stabilization, 30 minute after ischemia, and 60 minute after reperfusion. Western blotting was performed for evaluation of pAMPK at the end of the ischemia in the heart tissues.
The results demonstrated that hesperidin caused a significant reduction in ventricular ectopic beats (VEBs) number during ischemia and reperfusion phase (p The results of the study suggest that hesperidin has protective effects against I/R induced injuries and arrhythmias in isolated rat hearts that could be related to its effect on modulating of AMPK activity.

Several in vitro assays are used to determine Angiotensin Converting Enzyme (ACE) activity. The purpose of the present investigation, was developing a practical and extraction-free chromatographic method to determine ACE activity.
The method relies on UV-detection of Naphthoyl-glycine (NG), which is resulted from enzymatic hydrolysis of the synthetic substrate, Naphthoyl-glycyl-glycyl-glycine (NGGG), applying a reverse phase chromatographic separation. In this regard, experimental conditions for the assay such as Enzyme/Substrate (E/S) ratio and incubation time were optimized. Chromatographic separation was performed on a reverse phase C18 column (250 × 4.6 mm), using a mobile phase consisting of acetonitrile/water (20:80, v/v), pH = 5.0 with a flow rate of 2.0 mL/min and a detection wavelength of 280 nm.
The optimized Enzyme/Substrate (E/S) ratio and incubation time were 10 mU/nmol and 35 min respectively. The results indicated that the calibration curve was linear (r2 = 0.994) and the average recovery (n = 6) of NG was 99.5 ± 1.3% (mean ± RSD).
In this study, we introduced a method which is an efficient approach to determine ACE activity due to its sensitive, accurate, and reliable performance with great repeatability.

Tyrosinase is a key enzyme in melanin synthesis from tyrosine. To prevent or treat pigmentation disorders, tyrosinase inhibitors have been used increasingly for medicinal and cosmetic products. The aim of this study is to evaluate inhibitory effects of Urginea maritima (L.) Baker, Zhumeria majdae Rech.f. & Wendelbo and Physalis divaricata D.Don on mushroom tyrosinase.
The inhibitory activities of the hydroalcoholic extracts of plants against oxidation of L-DOPA (as a substrate) by mushroom tyrosinase were investigated. The amount of formed DOPAchrome was determined at 475 nm as optical density.
The extracts showed anti-tyrosinase activity weaker than positive control (Kojic acid). The inhibitory activity of tested plants: U.maritima, Z.majdae and P.divaricata against mushroom tyrosinase were 38.61, 29.70 and 25.74 % at 1.67 mg/mL, respectively.
The most tyrosinase inhibitory activity was seen for U.maritima. However more investigations on human tyrosinase, toxicological and clinical studies are needed to confirm its activity.

Analysis of aluminium (Al) in urine samples is required in management of a number of diseases including patients with renal failure. This work aimed to present dispersive liquid-liquid microextraction (DLLME) and ultrasound-assisted emulsification microextraction (USAEME) methods for the preconcentration of ultra-trace amount of aluminum in human urine prior to its determination by a graphite furnace atomic absorption spectrometry (GFAAS).
The microextraction methods were based on the complex formation of Al3 with 8-hydroxyquinoline. The effect of various experimental parameters on the efficiencies of the methods and their optimum values were studied.
Under the optimal conditions, the limits of detection for USAEME-GFAAS and DLLME-GFAAS were 0.19 and 0.30 ng mL−1, respectively and corresponding relative standard deviations (RSD, n=5) for the determination of 40 ng mL−1 Al3 were 5.9% and 4.9%.
Both methods could be successfully used to the analysis of ultra trace concentrations of Al in urine samples of dialysis patients.

A rapid stability-indicating RP-HPLC method for analysis of doxycycline in the presence of its degradation products was developed and validated.
Forced degradation studies were carried out on bulk samples and capsule dosage forms of doxycycline using acid, base, H2O2, heat, and UV light as described by ICH for stress conditions to demonstrate the stability-indicating power of the method. Separations were performed on a Perfectsil® Target ODS column (3-5µm, 125 mm×4 mm), using a mobile phase consisting of methanol-50 mM ammonium acetate buffer (containing 0.1% v/v trifluoroacetic acid and 0.1% v/v triethylamine, pH 2.5) (50:50 v/v) at room temperature. The flow rate was 0.8 mL/min.
The method linearity was investigated in the range of 25–500 µg/mL (r > 0.9999). The LOD and LOQ were 5 and 25 µg/mL, respectively. The method selectivity was evaluated by peak purity test using a diode array detector. There was no interference among detection of doxycycline and its stressed degradation products. Total peak purity numbers were in the range of 0.94-0.99, indicating the homogeneity of DOX peaks.
These data show the stability-indicating nature of the method for quality control of doxycycline in bulk samples and capsule dosage forms.

Two simple, precise and accurate multivariate calibration methods, partial least square (PLS) and principal component regression (PCR) have been applied for the simultaneous determination and dissolution profile evaluation of atorvastatin (ATV) and ezetimibe (EZT) in their binary mixtures and commercial tablets. Due to the closely overlapping spectral bands of the mentioned drugs, simultaneous determination without previous separation is not possible by conventional spectrophotometric methods. In the proposed methods (PLS and PCR) determination of chemicals was performed by the use of a full-spectrum multivariate calibration method.
The experimental calibration matrix was designed orthogonally with 16 samples composed of different mixtures of both compounds in related mediums. The simultaneous determination of ATV and EZT was accomplished in mixtures through recording the absorption spectra within a range of 210 to 300 nm.
The concentration of ATV and EZT were considered in the linear range, between 8 to 14 µg.ml-1. The specificity of the methods was evaluated by analyzing laboratory prepared mixtures of the mentioned drugs in specific proportions.
The applied methods were successfully employed in simultaneous spectrophotometric determination and dissolution profile evaluation of ATV and EZT in their prepared mixtures and pharmaceutical formulation.

Plant phenolic compounds are a main group of plant natural products and flavonoids are the largest and best studied natural phenols. These substances possess a series of biological properties and act on biological systems as antioxidants. In present research, the aim is to determine in vitro total phenolics content and antioxidant activity of the methanolic extracts of three Onobrychis species belonging to the family Fabaceae, namely O. sosnovskyi Grossh., O. viciifolia Scop. and O. melanotricha Boiss. Furthermore, an attempt was made to identify any correlations between total phenolic content of the extracts with their antioxidant activities
Total phenol and flavonoid contents of the extracts were determined by Folin-Ciocalteu and aluminum chloride methods, respectively. Antioxidant activity was evaluated by three different test systems, namely 2, 2-diphenyl-1-picrylhydrazyl free radical scavenging (DPPH), metal-chelation activity and ß-carotene/linoleic acid model.
Results indicated that O. viciifolia extract contains the highest total phenolic content (10.38 ± 0.33mg GAE/g of dry extract). However, the species are not remarkable different (P 0.55 to 0.98).
Our results showed that the examined Onobrychis extracts represent strong antioxidant activity; hence, they can be suggested as antioxidant agents for special use in future.

Palestine is a rich land with wild edible plants which used from the ancient times as food and medicine. Arum is a valuable genus of medicinal plants which is used in the ethnic medicine for treatment of cancer or consumed as integrated food. This work aimed to evaluate and compare the phytoconstituents, total phenols contents and free radical scavenging potential for Arum dioscoridis, Arum elongatum, Arum hygrophilum and Arum palaestinum a members of Palestinian flora.
Phytoconstituents screened by using standard analytical methods, total phenols determined by using Folin Ciocalteu's method and antioxidant activities were assessed by DPPH assay.
The crude extracts of Arum plant studied species revealed the presence of several biologically active phytochemicals with the highest quantity of saponin, alkaloid, phenols and flavonoids. For A. dioscoridis, A. elongatum, A. hygrophilum and A. palaestinum free radical scavenging activities were 6.7±0.75µg/ml, 19.9±0.63µg/ml, 9.9±0.49µg/ml, and 6.9±0.62µg/ml respectively, while the IC50 for Trolox was 4.8±0.39µg/ml as well as the total phenols contents for these species were 60.07 ±0.12, 27.49 ±0.32, 41.75±0.12 and 53.17±0.22 (mg GAE/g extract), ±SD respectively.
The antioxidant activities in the studied Arum plant species showed a marked correlation with their total phenols contents. A. dioscoridis and A. palaestinum had the highest antioxidant activities with high contents of total phenols and they can be used as perfect choices for manufacturing of pharmaceutical, cosmeticeuticals and nutraceutical formulations.

The suspension culture of Vigna radiata was selected for biotransformation of hydroquinone to its β-D-glucoside form (arbutin) as an important therapeutic and cosmetic compound.
The biotransformation efficiency of a Vigna radiata cell culture in addition to different concentrations of hydroquinone (6-20 mg/100 ml) was investigated after 24 hours in comparison to an Echinacea purpurea cell culture and attempts were made to increase the efficacy of the process by adding elicitors.
Arbutin was accumulated in cells and found in the media only in insignificant amounts. The arbutin content of the biomass extracts of V. radiata and E. purpurea was different, ranging from 0.78 to 1.89% and 2.00 to 3.55% of dry weight, respectively. V. radiata demonstrated a bioconversion efficiency of 55.82% after adding 8 mg/100 ml precursor, which was comparable with result of 69.53% for E. purpurea cells after adding 10 mg/100 ml hydroquinone (P>0.05). In both cultures, adding hydroquinone in two portions with a 24-hour interval increased the biotransformation efficiency. Different concentrations of methyl jasmonate (25, 50, and 100 µM) and chitosan (50 and 100 µg/ml) as elicitors increased the bio-efficiency percentage of the V. radiata culture in comparison with the flask containing only hydroquinone.
This is the first report of the biotransformation possibility of V. radiata cultures. It was observed the bioconversion capacity increased by adding hydroquinone in two portions, which was comparable to adding an elicitor.

Poisoning is a major cause of morbidity and mortality in children. Medicines and household cleaning products are responsible for the majority of cases. The aim of the present study is to analyze poisoning cases presenting to Tabriz children hospital. The present descriptive cross-sectional study, of all poisoning cases presenting to the children hospital during January 2014 to July 2015 and children from one to twelve years of age were included. The data was collected through referring to all parents of the children and using a questionnaire that included demographic and poisoning characteristic information. The demographic features included gender, age, place of residency and type of poisons was investigated. Also parents were interviewed by using structured questionnaire containing information on socio-demographic factors, parental smoking, parental education level, child's behavior, and storage practices of hazardous substances of caregivers inside homes. Children poisoning was common among low educated family with parental smoker and was higher for boys (59%) than girls (41%). However, there was no mortality. Poisoning was unintentional and most of the poisoned cases, 63 (61.7%) of the children involved were below the age of four years i.e., between 1 and 3 years of age. Pharmaceutical products were the commonest agents accounting for 50% of all cases; followed by pesticides (15.7%), poisonous mushrooms (13.7%), Petroleum products (10.8%) and household detergents (9.8%). Improvement in the socioeconomic status of parents and health education on proper/ safe storage of medicines, chemicals and household detergents will help in reducing the incidence of poisoning.

Plumbaginaceae plants family is a valuable natural insecticidal compound. This research focused on contact toxicity and chemical composition of essential oil obtained from Acantholimon scorpus. The essential oil from the aerial parts of A. scorpius was extracted by hydrodistillation method and tested for their toxicity against Oryzeaphilus mercator (Coleoptera: Silvanidae) and Tribolium castaneum (Coleoptera: Tenerbrionidae). Chemical compounds of the essential oil was analyzed by the gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID). The essential oil showed toxic effect on tested insects. This oil showed 90.0% mortality of O. mercator and 85.2% mortality of T. castaneum at a dose of 12 μl /l air after 48h of exposure. The constituents of this oil were identified, representing more than 82.9% of the total essential oil composition. Hexadecanoic acid, tetrahydrogeranyl acetone and oleic acid were the main compounds of the essential oil. According to the result the essential oil of A. scorpius showed a noticeable insecticidal activity in contact toxicity model.