A simultaneous liquid chromatographic analysis of niclosamide and bicalutamide in rat plasma by protein precipitation extraction.

OBJECTIVE: The aim of the study was to develop a simple, sensitive and cost-efficient bioanalytical method for the simultaneous determination of niclosamide and bicalutamide in rat plasma with appropriate validation. MATERIAL AND METHODS: Real-time estimation was carried out using Ultra Flow Liquid Chromatography. The solvent system consisting of 20mM sodium acetate buffer at pH 4.0 and acetonitrile (65: 35% v/v) is used as the mobile phase. The analytes were extracted by protein precipitation with acetonitrile and separated on an Eclipse plus C18 Column (25cm×5cm×4.6µm) with L1 packing in isocratic mode with a flow rate of 1ml/min at 254nm. The developed method was validated on the basis of the bioanalytical guidelines of the US American FDA. RESULT: The retention times of niclosamide and bicalutamide were 4.19min and 8.61min, respectively, with a running time of 15minutes. The calibration ranges are 50-600ng/ml for niclosamide and 100-1200ng/ml for bicalutamide. Regression equations for niclosamide and bicalutamide were y=55746x - 1E+06 and y=22051x-576888 with regression coefficient (R2) 0.9952 & 0.9982 using the unweighted and weighted linear regression with a weighting factor of 1/xo, 1/x, 1/√x, and 1/x2. The accuracy of niclosamide and bicalutamide were 46.01ng/ml to 584.10ng/ml and 82.30ng/ml to 1149.13ng/ml, respectively. The percentage recoveries for niclosamide and bicalutamide were 86.51-90.29% & 88.64-93.99%, respectively. CONCLUSIONS: The result of the present study show that the method developed is a simple, fast and precise method and is used in bioavailability and bioequivalence studies as well as during routine therapeutic drug monitoring of niclosamide and bicalutamide.

Formulation and evaluation of semisolid dosage forms of an anti-inflammatory drug.

Curcumin is one of the commonly used dietary supplements with wide pharmacological activities but the main hindrance in the commercial exploitation of curcumin is the issue with its solubility and stability. Hence, the aim of the present study was to formulate curcumin silver nanoparticles (CSNP) by chemical reduction method and to evaluate its solubility, stability as well as diffusion properties. The CSNP was combined with potent anti-inflammatory drug Diclofenac sodium (DS) to prepare gels (F1-F7), cream (F8) and ointment (F9). The DS-CSNP was subjected to in vitro and in vivo anti-inflammatory activity by albumin denaturation method and carrageenan-induced paw oedema method using albino rats, respectively. Results of the present study revealed that CSNP possess good solution stability in different pH solutions compared to pure curcumin, and the particle size as well as zeta potential were found to be 115 nm and - 5.69 mV, respectively for CSNP. Based on the results of in vitro release study and in vitro anti-inflammatory activity, formulation F4, F5, F9 and marketed product were subjected for in vivo anti-inflammatory activity. Among the three formulations, formulation F9 showed the maximum inhibition of the oedema (82.25%) at the end of 90 min followed by F5 and F4. In addition, formulations F4, F5, and F9 exhibited better in vivo anti-inflammatory activity in comparison with the marketed product which might be due to synergistic effect of the combination of curcumin silver nanoparticles and DS. In conclusion, CSNP-incorporated DS semisolid preparations were stable and can yield promising anti-inflammatory activity compared to marketed formulation; hence, these formulation can be exploited commercially in the preparation of anti-inflammatory dosage forms.

Evaluation of phenolic content and antioxidant potency in various parts of Cassia auriculata L.: a traditionally valued plant.

Presence of polyphenolic content in various part of the plant exhibit wide pharmacological activities including antioxidant activity. The present study was designed to evaluate the phenolic contents (total phenols, flavonoid and tannins) and antioxidant properties of ethanolic extracts of flower, leaf, pod, bark and root obtained from Cassia auriculata. Ethanolic extracts of various parts of C. auriculata obtained by sonication extraction techniques are studied for their phenolic contents and DPPH (2,2-diphenyl-1-picrylhydrazine) radical scavenging assay as well as total antioxidant assays using UV visible spectrophotometer. Among the various parts of the plant studied, bark showed significant content of phenolics, flavonoids and tannins followed by the root, leaf, flower and pod. Even bark extract exhibited highest antioxidant capacity in DPPH assay followed by root, leaf, flower and pod with a value of 766.7, 679.3, 644.9, 572.5 and 474.7 mg vitamin C equivalent antioxidant capacity (mg VCEAC)/sample, respectively. In addition, mg VCEAC values obtained from the total antioxidant assay was in the increasing order of bark > root > leaf > flower > pod. Moreover, a strong correlation was also found between phenolic contents and antioxidant values indicating their influence in the found antioxidant activity, hence the bark extract can be employed as an ideal candidate for herbal based pharmaceutical product. Results of the present study also emphasize variation in the chemical composition as well as biological activity ensuring the importance of proper selection of particular part of the plant to evaluate their therapeutic potency.

The free radical scavenging and antioxidant activities of pod and seed extract of Clitoria fairchildiana (Howard)- an underutilized legume.

We evaluated the phenolic content and antioxidant capacities of pod and seed extracts (in methanol, ethanol, and water) of an underutilized legume, Clitoria fairchildiana (Howard). The antioxidant capacity of the extracts was determined using the ferric reducing antioxidant potential assay, and the free radical-scavenging capacity was evaluated using 2,2-diphenyl-1-picrylhydrazyl radical-scavenging and ABTS assays. In addition, the total flavonoids, flavonols, and tannin contents were also determined. Overall, the methanol extracts of the pod contained high concentration of phenolics and showed high antioxidant capacities compared to seed extracts. In addition, a positive correlation was found between total phenol and tannin versus antioxidant capacity. Results of the present study indicate pods and seeds of C. fairchildiana to possess rich amount of natural antioxidants, and can be further explored for their possible use as a natural additive in food or in pharmaceutical industries.

Influence of sonication treatments and extraction solvents on the phenolics and antioxidants in star fruits.

The present study was conducted to examine the effects of sonication treatments (time intervals of 0, 15, 30, 45 and 60 min.) on phenolics and other antioxidant compounds in starfruits extracted in methanol and water. Overall, methanolic extracts exhibited significantly higher extractability, percentage inhibition of DPPH radicals, ferric reducing antioxidant property (FRAP) value, antioxidant capacity, flavonoids, total phenolics and tannins (p < 0.05) compared to control (0 min) and aqueous extracts. Methanolic extract obtained after 30 min of sonication proved to be the best treatment with regard to various parameters evaluated. Results of the present study clearly indicated sonication treatments to be effective in enhancing the antioxidant compounds in starfruit extracts and could be further explored for commercial purposes to benefit the consumers.

Influence of sonication on the phenolic content and antioxidant activity of Terminalia catappa L. leaves.

AIM: This study was designed to evaluate the phenolic content and antioxidant activity of ethanolic extracts from T. catappa leaves obtained by different intervals of sonication. METHODS: Three commonly used methods were followed to evaluate phenolic content and four in vitro methods like 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging assay, ferric reducing antioxidant potency (FRAP), and total antioxidant capacity assays for measuring the antioxidant activities. Antioxidant values of these assays were expressed in terms of milligrams vitamin C equivalent (VCE) antioxidant activities. RESULTS: This study showed that extract obtained with 40 minutes of sonication possessed significant (P < 0.05) polyphenolic contents compared to 20 and 60 minutes sonication and control (24 hour maceration). Moreover, sonication of T. catappa leaf above 40 minutes was found to be unsuitable for extracting out phenolic contents. Even the results of antioxidant assays showed that 40 minutes of the sonicated extract exhibited significant (P < 0.05) VCE values compared to extracts obtained at different intervals of sonication and control. CONCLUSIONS: In sonication extraction method 40 minutes is an ideal time to obtain extract enriched with high polyphenolic content with good antioxidant activity from T. catappa leaves.