Iranian Journal of Pharmaceutical Sciences
Volume:15 Issue: 4, Autumn 2019

In Indonesia, Caesalpinia pulcherrima (L) Swartz is widely grown as an ornamental, whereas it is used as a medicinal herb in Mexico as an alternative treatment for dental and oral infections. At present, only fruit and root parts of the plant are used. The aim of this study was is evaluate the anti-microbial activity of ethanol extract of fruit, leaves, flowers and stems against microbes that cause dental and oral infections and determination of the total flavonoid and total phenolic contents of C. pulcherrima (L) Swartz. Porphyromonas gingivalis, Sterptococcus mutans, Enterococcus faecalis and Candida albicans are microbes that cause dental and oral infections. All parts of the plant were extracted by maceration using 70% ethanol for 24 hours at 150-300C, and extracts were concentrated using a reflux method and dried over a water bath. The total flavonoid contents were determined using the aluminium chloride colorimetric assay at 415 nm. The total phenolic content was determined using Folin–Ciocalteu reagent in a 1:4 ratio at a wavelength of 750 nm using a microplate reader. Antimicrobial activity was determined by the diffusion method. The total flavonoid content was expressed as milligram quercetin equivalent (QE) per gram of plant extract. The highest total flavonoid content per gram of plant extract was found in the flowers (10.06 ± 0.08 mg QE/g), and the lowest total content was found in the leaves (0.34 ± 0.02 mg QE/g). The highest phenolic content was also found in the flowers (498.52 ± 1.96 mg gallic acid equivalent [GAE]/g), and the lowest total phenolic content was detected in the leaves (216.76 ± 1.00 mg GAE/g). A 100% crude ethanol extract of the flower and fruit parts exhibited antimicrobial activity against bacteria P. gingivalis, S. mutans, E. faecalis and C. albicans) that cause dental and oral infections. A 2% crude flower extract showed the highest antimicrobial activity against P. gingivalis (14.49 ± 0.465 mm), S. mutans (9.02 ± 0.607 mm), E. faecalis (9.67 ± 0.297 mm) and C. albicans (19.44 ± 0.207 mm). The ethanol extract of the flowers had the highest contents of total flavonoids and total phenols and the most antimicrobial activity against the studied compared to the other parts. This results of the present study revealed valuable information and also support the continued sustainable use of C. pulcherrima (L) Swartz in a traditional system of medicine

In recent years, cleaning validation has achieved a position of increasing in the pharmaceutical industry. It provides assurance to the cleaning procedure that ensures equipment is consistently cleaned from the product, detergent and microbial residues to an acceptable level to avoid cross-contamination and adulteration of drug product with other active ingredients. The aim of this study was to demonstrate the applicability of reversed-phase high-performance liquid chromatography coupled with UV detector (RP-HPLC-UV) method for determining the residues of Diclofenac sodium in cleaning control swab samples from equipment surfaces after manufacturing of Diclofenac sodium injection (75mg/3ml) in order to control a cleaning procedure. Diclofenac sodium was evaluated as the worst case. This API is sparingly soluble in water and adherent to surfaces. The acceptable residue limit (ARL) of Diclofenac sodium was calculated (0.75 µg/cm²). The analytical method was validated with respect to system suitability test, specificity, linearity-range, accuracy, Repeatability, intermediate precision, limit of detection (LOD) and quantitation (LOQ). These studies were performed in accordance with established guidelines, International Conference on Harmonisation ICH Q2 (R1). The precision of the swabbing procedure and stability of Diclofenac sodium standard solutions were also investigated. The swab sampling method was developed and optimized in order to obtain a suitable recovery (˃80%) from stainless steel surfaces 316L. Alpha Texwipe® TX761 polyester Swabs were moistened with diluent (mobile phase) a mixture of Acetonitrile-Water-Orthophosphoric Acid 85 % (600:400:1, v/v/v) (pH 2.5* ± 0.2). The HPLC method was developed in isocratic mode using Hypersil OctaDecylSilyle ODS C18 (250 × 4.6 mm, 5 μm) column at 25°C. At a flow rate of 1.2 ml/min, an injection volume of 20 μl. Detection was carried out at 235 nm. The retention time of Diclofenac sodium was 4.8 min. The calibration curve was linear (the coefficient of determination R²= 0.9988) over a concentration range 0.05 μg/ml – 12.5 μg/ml. The intra-day, inter-day precision and precision of the swabbing procedure expressed as relative standard deviation were below 5%. The limit of detection and quantitation were 0.014 μg/ml and 0.05 μg/ml respectively. The average recovery of the swabbing method obtained was 87.80 %, when two swabs moistened were used. It is evident that this proposed validated RP-HPLC-UV method with the appropriate swabs Texwipe® TX761 procedure could be applicable for cleaning validation to detect traces levels of Diclofenac sodium residues on pharmaceutical manufacturing equipments.

Cytarabine is the drug of choice for treatment of leukemia. However, many formulations have been prepared, due to certain limitations they could not prove to be effective ones, therefore magnetic microspheres are formulated as they minimize the Reticuloendothelial clearance and target site specificity can be increased. The current study aimed to utilize nanotechnology to develop magnetic microspheres. Magnetic microspheres of cytarabine were prepared using two polymers are chitosan and sodium alginate by the continuous solvent evaporation method. Optimization was done and nine different formulations were prepared. Particle sizes, encapsulation efficiency, magnetic responsiveness, in vitro release of all the formulations were determined. The studies demonstrated that F7 was the best formulation and drug can thus be reached to the targeted site as magnetic responsiveness was also good. Novel magnetic microspheres were developed with less Reticuloendothelial clearance and target site specificity; however, further clinical investigations are necessary to evaluate its therapeutic effectiveness.

In this study, extraction of amoxicillin (AMOX) ions from aqueous solution by liquid-liquid extraction (LLE) was investigated. The LLE process by response surface methodology (RSM) was carried out based on central composite design (CCD). The organic phase consisted of extractant and toluene as diluent. In this process, triethylamine (TEA), di-2-ethylhexyl phosphoric acid and mono-2-ethylhexyl phosphoric acid mixture (MDEHPA) were used as extractant. The process parameters such as extractant concentration, feed acidity, extraction time and effect of diluents were optimized. The contours and 3D response surfaces of AMOX extraction efficiency were obtained. The optimum conditions for the AMOX extraction using RSM for TEA were: feed pH 2.7, extractant concentration 12.9 %( V/V) and extraction time 19 min and for MDEHPA: feed pH 11, extractant concentration 12.3 %( V/V) and extraction time 21 min. At these condition the maximum AMOX extraction was found to be 93.5 % and 96 % using TEA and MDEHPA, respectively.

A New Stability Indicating RP-HPLC Method Development and Validation for the Simultaneous Estimation of Dolutegravir and Rilpivirine in Bulk and its Dosage Forms. Abstract The objective of the work is to develop and validate a new, simple, highly sensitive, stability indicating RP-HPLC method for simultaneous estimation of Dolutegravir and Rilpivirine in bulk and its dosage forms. The method was developed on a reversed-phase Thermosil C18 (4.6 × 150 mm, 5µm) column with an isocratic elution. The Mobile phase ratio was Acetonitrile: Phosphate Buffer pH 3.5 (45:55 % v/v). Detection was done by UV-Spectroscopy at a detection wavelength of 260 nm. The flow rate was 0.8 ml/min. The mobile phase was used as a diluent. The Injection volume was 10μl. The analytical procedure was validated as per ICH guidelines. The retention time for Dolutegravir and Rilpivirine in the standard solution having the concentration of 100 μg/ml of Dolutegravir and 50 μg/ml of Rilpivirine were observed to be around 2.427 min and 4.436 min. respectively. The purity percentage values of Dolutegravir and Rilpivirine were 99.22 % w/v and 99.81 % w/v respectively. System suitability parameters were calculated and found within the acceptance criteria. The proposed method was found to have a high degree of precision and reproducibility. Calibration plots were linear (r2> 0.999) over the concentration range of 80 - 120 μg/ml for Dolutegravir and 30 - 70 μg/ml for Rilpivirine. The recovery percent were within the acceptance criteria of 98 – 102 % for Dolutegravir and Rilpivirine. The LOD was 0.044 µg/ml for Dolutegravir and 0.060 µg/ml for Rilpivirine. The LOQ was 0.134 µg/ml for Dolutegravir and 0.183 µg/ml for Rilpivirine. The method represents a fast-analytical procedure and stability indicating analytical method for the simultaneous estimation of Dolutegravir and Rilpivirine in bulk and its dosage forms. No interference from any component of pharmaceutical dosage form was observed. Validation studies revealed that the method is specific, rapid, reliable, and reproducible. The method is amenable to the routine analysis of large numbers of samples with good precision and accuracy.

In this study, we aimed to determine VEGFR-2, EGFR and PDGFR-β tyrosine kinase inhibitory activities of some pyrrolo[2,3-d]pyrimidine derivatives previously synthesized and showed potent cytotoxic and apoptotic effects against several cancer cell lines by our group and to evaluate the relationships between inhibitory activities and binding properties of the active compounds by molecular docking studies. VEGFR-2, EGFR ve PDGFR-β tyrosine kinases inhibitory activities of the tested compounds were determined using KDR Kinase Enzyme System Analysis Kit (Promega, #V2681), EGFR Kinase Enzyme System Analysis Kit (Promega, #V3831) and PDGFR-β Kinase Enzyme System Analysis Kits (Promega, #V3731) according to the manufacturer’s instructions. The molecular docking studies were performed using Autodock vina program. Compounds 9a, 9b and 11b exhibited the weak inhibitory activities against VEGFR-2, EGFR and PDGFR-β, respectively. Molecular docking studies showed that one or two hydrogen bonding interactions were found between compounds 9a, 9b, 11b and VEGFR-2, EGFR, PDGFR-β tyrosine kinases. Biological activity and molecular docking results revealed that interactions of compounds with target protein active sites are not enough to obtain potent RTK inhibitory activity. It is necessary to design some compounds showing more interactions with the target proteins to obtain better activity results.

Vincristine (VCR) is potential anti-cancer drug that is highly toxic for renal tissue. This study aimed to evaluate the protective effect of alcoholic extract of saffron stigma against vincristine renal toxicity in male rats. A total number of 50 rats were randomly divided into 10 groups. Different doses of VCR (0.25, 0.5 and 0.75mg/kg) and saffron (0.5 and 1mg/kg) were used to treat vincristine-induced renal toxicity in rats. Serum level of renal biomarkers, creatinine (Cr), uric acid and blood urea nitrogen (BUN) were measured using specific kits at the end of the experimental period. Serum total antioxidant capacity (TAC) and malondialdehyde (MDA) values were measured using Ferric reducing antioxidant of power (FRAP) and Thiobarbituric acid reaction (TBAR) methods, respectively. Administration of VCR to the experimental rats caused severe renal injury with significant increase in the levels of creatinine (Cr), uric acid and blood urea nitrogen (BUN). VCR administration (at concentration of 0.75mg/kg) also significantly increased the mean level of MDA (0.51±0.021 nmol/ml; p<0.001), while TAC value was declined significantly (243.17±16.24 μmol/l; p<0.001). These effects were dose-dependently. Treatment with saffron extract decreased the level of Cr, BUN, and MDA values in VCR-exposed rats with a significant enhancement in serum TAC content. This effect was notable for rats that received 1mg/kg plant extract. Administration of saffron, especially at higher concentration, can reduce VCR-induced renal toxicity, total antioxidant depletion and lipid peroxidation, possibly due to its antioxidative properties.

Hypericum species are known to be used in traditional therapies. H. scabrum L. is one of the Hypericum species distributed in Turkey. In this study, we evaluated the effects of Hypericum scabrum essential oil inhalation on spatial memory in scopolamine-induced amnesic rats. The essential oil was characterized by GC-FID and GC-MS system. Male wistar rats were divided into 6 groups: control; scopolamine-alone treated; diazepam-treated (positive control); tramadol-treated (positive control); scopolamine with Hypericum scabrum essential oil 1%-treated; and scopolamine with Hypericum scabrum essential oil 3%-treated group. All rats were tested behaviorally to evaluate spatial memory peformances in Y-maze task and radial-arm maze task. The most abundant component of the essential oil was identified as α-pinene (51.3%). Hypericum scabrum essential oil application significantly improved spatial memory as compared to scopolamine-alone treated rats as evidenced by increases in spontaneous alternation behavior in Y-maze task and decreases in the number of working memory errors and reference memory errors in the radial-arm maze task in this study. Hypericum scabrum essential oil inhalation could be used as a complementary therapy to reduce memory impairments in the patients with dementia and related diseases.