Arjun Patra

Experimental techniques for screening of antiosteoporotic activity in postmenopausal osteoporosis.

Abstract Postmenopausal osteoporosis, a silent epidemic, has become a major health hazard, afflicting about 50% of postmenopausal women worldwide and is thought to be a disease with one of the highest incidences in senile people. It is a chronic, progressive condition associated with micro-architectural deterioration of bone tissue that results in low bone mass, decreased bone strength that predisposes to an increased risk of fracture. Women are more likely to develop osteoporosis than men due to reduction in estrogen during menopause which leads to decline in bone formation and increase in bone resorption activity. Estrogen is able to suppress the production of proinflammatory cytokines like interleukin (IL)-1, IL-6, IL-7 and tumor necrosis factor (TNF-α). This is why these cytokines are elevated in postmenopausal women. In this review article we have made an attempt to collate the various methods and parameters most frequently used for screening of antiosteoporotic activity in postmenopausal osteoporosis. Pertaining to ovariectomized animal model, this is the most appropriate model for studying the efficacy of different drugs to prevent bone loss in postmenopausal osteoporosis.

Estrogenic activity of Punica granatum L. peel extract

Abstract Objective To evaluate the estrogenic effect of alcoholic extract of Punica granatum peel (ALPG) in ovariectomized (OVX) rats. Methods Female wistar rats were divided into sham control and ovariectomized (OVX), OVX rats receiving standard drug, raloxifene (5.4 mg/kg) and groups treated with 500 mg/kg and 750 mg/kg of ALPG daily for 90 days. The vaginal cornification, uterine weight, bone loss, biomechanical, biochemical and histopathological observation were carried out to ascertain the effect of test drug in post menopausal syndrome. Results The experimental animals treated with the ALPG showed dose dependent activity. The significant increase in uterine weight, femur BMD, femur hardness was observed. In addition, increased levels of calcium and phosphorus in serum and significant decrease in urine, were observed as compared to control OVX group. The histopathological results also confirm the protective effect of ALPG. Conclusions The present findings strongly suggest that the peel of P. granatum possess potent estrogenic activity in ovariectomized rats and substantiate the ethnic use in treatment of postmenopausal osteoporosis.

Stability Indicating High Performance Thin Layer Chromatographic Method for the Determination of Tramadol Hydrochloride in Pharmaceutical Formulation

The proposed stability indicating method for the determination of tramadol hydrochloride by high-performance thin-layer chromatography in the pharmaceutical formulations was found to be specific, precise, and validated. The stationary phase employed was a precoated silica gel 60 F254 aluminum TLC plate. Several mobile phase combinations of varying polarity of solvent were tried for the method development, as well as for the resolution of degradation products from the parent densitogram of drug. Finally, the mobile phase containing a mixture of ethyl acetate:methanol:ammonia (9:0.8:0.5 v/v/v) was found to be satisfactory for the resolution of degradation products from the parent drug. Densitometric analysis of tramadol hydrochloride was carried out in the reflectance–absorbance mode at 271 nm. The Rf value of the drug was obtained at 0.64 ± 0.02 with sharp symmetrical peak. The degraded products formed under acidic, oxidative, and an alkali conditions were strongly retained. Linear relationships between concentration of analyte and corresponding peak area were observed over the range of 1000–6000 ng at the selected wavelength with an R2 value of 0.999 ± 0.0007. The validation for specificity, precision, robustness, and recovery of the method was also performed. The present method effectively separated the tramadol hydrochloride from its degradation products.

Simultaneous estimation of genistein and daidzein in Pueraria tuberosa (Willd.) DC by validated high-performance thin-layer chromatography (HPTLC) densitometry method

ABSTRACT The present study was performed to estimate the concentration of genistein and daidzein in ethanol extract of tubers of Pueraria tuberosa (Indian kudzu or Vidarikanda) and its various fractions (n-hexane, ethyl acetate, n-butanol, and aqueous) by high-performance thin-layer chromatography (HPTLC). The separation of bioactive compounds was performed using mobile phase, toluene:ethyl acetate:acetone:formic acid (20.0:4.0:2.0:1.0) and detected at wavelength 269 nm. The method was validated for linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), etc. by International Conference on Harmonization guidelines. The calibration range was found to be 100–600 ng/band for both the bioactive compounds. Daidzein was separated with an Rf value of 0.39 ± 0.02 and genistein with an Rf value of 0.54 ± 0.02. Average recovery was 99.96 and 99.90% for genistein and daidzein, respectively. The LOD and LOQ were 14.786 and 44.805 ng, respectively, for genistein, and 9.607 and 29.114 ng, respectively, for daidzein. Both the phytoconstituents were found in ethanol extract and its ethyl acetate fraction only. The developed HPTLC method was simple, precise, robust, specific, rapid, and cost effective and could be used for quality control analysis and quantification of genistein and daidzein in different herbal formulations containing the plant species. GRAPHICAL ABSTRACT

Pharmacognostical standardization of leaves of Xanthium strumarium Linn.

Abstract Xanthium strumarium L. (Compositae) (Hindi: Chota-gokhru), is a gregarious weed found abundantly throughout India. The whole plant is used as a diaphoretic, sedative, sudorific, diuretic and sialagogue. The ethanolic extract of the leaves exhibits significant anti-inflammatory, analgesic, antitrypanosomal and anti-microbial effects. The present study deals with the pharmacognostical evaluation of leaves of Xanthium strumarium Linn. Macromorphology and microscopy (transverse section, powder microscopy and quantitative microscopy) were studies to establish the salient diagnostic features. The preliminary phytochemical analysis and thin layer chromatography has also been performed. The results of this study could be useful in setting some diagnostic indices for identification and preparation of the monograph of the plant.

Formulation and evaluation of mixed polymeric micelles of quercetin for treatment of breast, ovarian, and multidrug resistant cancers

Background Quercetin (QCT), a naturally occurring flavonoid has a wide array of pharmacological properties such as anticancer, antioxidant and anti-inflammatory activities. QCT has low solubility in water and poor bioavailability, which limited its use as a therapeutic molecule. Polymeric micelles (PMs) is a novel drug delivery system having characteristics like smaller particle size, higher drug loading, sustained drug release, high stability, increased cellular uptake and improved therapeutic potential. In the present study, we have formulated and characterized mixed PMs (MPMs) containing QCT for increasing its anticancer potential. Methods The MPMs were prepared by thin film hydration method, and their physicochemical properties were characterized. The in vitro anticancer activity of the MPMs were tested in breast (MCF-7 and MDA-MB-231, epithelial and metastatic cancer cell lines, respectively), and ovarian (SKOV-3 and NCI/ADR, epithelial and multi-drug resistant cell lines, respectively) cancer. Results The optimal MPM formulations were obtained from Pluronic polymers, P123 and P407 with molar ratio of 7:3 (A16); and P123, P407 and TPGS in the molar ratio of 7:2:1 (A22). The size of the particles before lyophilization (24.83±0.44 nm) and after lyophilisation (37.10±4.23 nm), drug loading (8.75±0.41%), and encapsulation efficiency (87.48±4.15%) for formulation A16 were determined. For formulation A22, the particle size before lyophilization, after lyophilization, drug loading and encapsulation efficiency were 26.37±2.19 nm, 45.88±13.80 nm, 9.01±0.11% and 90.07±1.09%, respectively. The MPMs exhibited sustained release of QCT compared to free QCT as demonstrated from in vitro release experiments. The solubility of QCT was markedly improved compared to pure QCT. The MPMs were highly stable in aqueous media as demonstrated by their low critical micelle concentration. The concentration which inhibited 50% growth (IC50) values of both micellar preparations in all the cancer cell lines were significantly less compared to free QCT. Conclusion Both the MPMs containing QCT could be used for effective delivery to different type of cancer and may be considered for further development.

Antioxidant and anticancer activities of green synthesized silver nanoparticles using aqueous extract of tubers of Pueraria tuberosa

Abstract In the present study, we have synthesized silver nanoparticles (AgNPs) using Pueraria tuberosa aqueous extract (PTAE) in a straightforward manner without involvement of toxic chemicals. Various reaction parameters (reaction time, AgNO3 concentration, pH, reaction temperature and PTAE concentration) were optimizeded for the synthesis of AgNPs through visual observation of colour change and absorption peak by UV–Vis spectroscopy. The green synthesized AgNPs were analyzed by DLS, FTIR, SEM, TEM, EDS and XRD for their bio-physical characteristics. The AgNPs were screened for their antioxidant activity by determining total antioxidant capacity (TAC) and anticancer potential by MTT assay. The average particle size of AgNPs was 162.72 ± 5.02 nm with zeta potential of –30.14 ± 2.08 mV. The spherical shape of the AgNPs was confirmed by SEM and TEM. FTIR spectra showed the involvement of different phytoconstituents as capping and stabilizing agents in the synthesis of AgNPs. The TAC of the optimized synthesized AgNPs was more than PTAE. The IC50 of synthesized AgNPs against MCF-7, MDA-MB-231, SKOV-3, U-87 and NCI/ADR cell lines was 3.859, 1.128, 29.36, 6.053 and 25.49 µg/ml, respectively. The green synthesized AgNPs has potential for use in the treatment of different types of cancer.