Suresh Bhojraj’s research while affiliated with JSS College of Pharmacy and other places

A simple and validated liquid chromatographic–mass spectrometric method (LC-MS) for amlodipine in human plasma was quantifi ed using LC-MS (ESI). Chromatography was performed on a C₁₈ analytical column, the mobile phase used was Acetonitrile-10mM Ammonium acetate in the ratio of 90:10%v/v and the retention times were 0.829 and 1.281 min for azithromycin (Internal standard) and amlodipine respectively. The ionization was optimized using ESI (+) and enhanced selectivity was achieved. The method is validated as per FDA guidelines. The analyte was shown to be stable over the timescale of the whole procedure. The pharmacokinetic parameters such as peak plasma concentration (C_max), Time to peak Concentration (t_max), Area under the plasma concentration-time curve (AUC_0-t & AUC_0-∞), elimination rate constant (K_eli), Elimination half-life (t_½) were calculated. Log transferred values were compared by Analysis ofVariance (ANOVA) followed by classical 90% confidence interval for C_max AUC0-t.and AUC_0-∞ and was found to be within the range. These results indicated that the Test and Reference formulation is bioequivalent.

A sensitive and reproducible high performance liquid chromatography (HPLC) method has been developed and validated for the quantification of metolazone in human plasma, after solid phase extraction (SPE). A Good resolution was achieved on a reverse-phase LichroCART Purospher® C₁₈ column using the mobile phase acetonitrile–0.5% triethylamine (35:65) in isocratic elution with a total run time of 15 min. The analyte, metolazone, was detected by using high performance liquid chromatography with the support of photo diode array detector. Limit of detection and Lower limit of quantification was found to be 1 and 2.5 ng/mL. The present method was successfully applied in the pharmacokinetic study of metolazone in human plasma.

A sensitive and reproducible high performance liquid chromatography (HPLC) method has been developed and validated for the quantification of prochlorperazine sustained release tablets in human plasma, after solid phase extraction (SPE). Best chromatographic resolution wasachieved on a reverse-phase Phenomenex C₁₈ column using the mobile phase consisted of a mixture of 20 mM disodium hydrogen ortho phosphate–acetonitrile (95:5) in an isocratic elution with a total run time of 12 min. Linear plot was obtained in the concentration range of 15–300 ng/ml (r² = 0.99). Lower limit of quantification (LLOQ) was found to be 15 ng/ml. Average recovery of the analyte was found to range from 98.25 to 99.13% in plasma at the concentrations of 45, 150 and 270 ng/ml. The intra and inter-day relative standard deviations of low quality control (LQC), medium quality control (MQC) and high quality control (HQC) of prochlorperazine were found to be 2.63, 3.25, 2.83 and 3.57, 5.88 and 3.78 respectively. The present method was successfully applied in the pharmacokinetic study of prochlorperazine in human plasma.

Four successive extracts of the whole plant of Enicostemma axillare (E. axillare), were examined for in vitro antioxidant activity using nine different methods. In the 2,2 -azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid) diammonium salt (ABTS) method, all the four extracts of E. axillare showed potent antioxidant activity with IC 50 values ranging from 13.26 to 24.36 µg/ml. The chloroform extract has shown potent antioxidant activity in H 2 O 2 , nitric oxide, and hydroxyl radical using the deoxyribose and lipid peroxidation methods, with IC 50 values of 16.99 ± 0.38, 60.66 ± 0.30, 25.06 ± 0.12, and 94.66 ± 2.40 µg/ml, respectively. Potent activity was also observed for the petroleum ether extract with the deoxyribose, p-nitroso dimethyl aniline (p-NDA), and H 2 O 2 methods and for the ethyl acetate extract with the H 2 O 2 and nitric oxide methods. All extracts showed moderate total antioxidant capacity using the phosphomolybdenum method. The activity was not correlated with the total phenol content of the extracts.

The methanol extract of Careya arborea bark (MECA) was tested for antioxidant and hepatoprotective activity in Ehrlich ascites carcinoma (EAC) tumor-bearing mice. Tumor control animals inoculated with EAC showed a significant alteration in the levels of antioxidant and hepatoprotective parameters. The extract treatment at 50, 100 and 200 mg/kg body weight doses given orally caused a significant reversal of these biochemical changes towards the normal in serum, liver and kidney when compared to tumor control animals indicating the potent antioxidant and hepatoprotective nature of the standardized extract.

The present study was carried out to evaluate Ficus glomerata extracts for antioxidant and hepatoprotective properties. The methanol extract of the bark of F. glomerata showed potent in vitro antioxidant activity when compared to the root methanol extract. In the in vivo studies, the CCl(4) treated control rats showed a significant alteration in the levels of antioxidant and hepatoprotective parameters. The methanol extract of the bark when given orally along with CCl(4) at the doses of 250 and 500 mg/kg body weight showed a significant reversal of these biochemical changes towards the normal when compared to CCl(4)-treated control rats in serum, liver and kidney. The results were comparable to those observed for standard sylimarin. Histological studies also confirmed the same. The results indicated the potent hepatoprotective and antioxidant nature of the bark extract.

The Argyreia cymosa bark extracts were subjected to in vitro antioxidant activity with different methods. The petroleum ether extract has shown antioxidant activity in ABTS, nitric oxide, hydroxyl radical (by p-NDA) and lipid peroxidation methods. The ethyl acetate extract has shown antioxidant activity in DPPH, H(2)O(2) and hydroxyl radical (by deoxyribose) scavenging methods.

The effect of ethanol extract of Phyllanthus maderaspatensis (PME), a popular south Indian dietary supplement, was studied for its chemoprotective property on adriamycin (ADR)-induced toxicity and oxidative stress in mice. Adriamycin toxicity was evaluated biochemically by measuring the serum concentration of lactate dehydrogenase (LDH) and creatinine phosphokinase (CPK). Genotoxicity was evaluated by measuring the frequency of micronucleated polychromatic erythrocytes (MNPCEs) in bone marrow cells. Oxidative stress in the heart tissue was estimated by measuring the glutathione (GSH) levels in the homogenate. The treatment of mice with different doses of PME (400 mg/kg and 600 mg/kg body weight, p.o.,) for 7 days before the administration of a single i.p. dose of ADR (15 mg/kg) exhibited significant protection in a dose-dependent manner. The results clearly indicate that PME has a protective effect against ADR-induced toxicity, as revealed by the decrease in the concentrations of LDH, CPK, and the frequency of MNPCEs. The increased levels of GSH are indicative of the antioxidant property of PME.

Herbex-kid (HK), a polyherbal formulation was evaluated in various experimental allergic models of Type I hypersensitivity reactions. Compound 48/80 (C 48/80) has been shown to induce rat mesentery mast cell degranulation and HK (1.07, 10.75 and 107.5 mg ml(-1)) inhibited the mast cell degranulation in a dose dependent manner. HK (1.07, 10.75 and 107.5 mg kg(-1); p.o.) showed dose-dependent protection against C 48/80 induced systemic anaphylaxis in male Balb/C mice. In active anaphylaxis model, male Wistar rats orally administered with 10.75 and 107.5 mg kg(-1) of HK showed significant (P < 0.01) protection against mast cell degranulation, while in passive anaphylaxis model, only at 107.5 mg kg(-1) showed significant (P < 0.01) reduction in mast cell degranulation. HK at all dose levels was able to significantly decrease the time spent in nasal rubbing in Wistar rats sensitized to ovalbumin, while only at 107.5 mg kg(-1) it showed significant (P < 0.01) reduction in number of sneezes. In C 48/80-induced skin itch model, all dose levels of HK significantly (P < 0.001) decreased the time spent in itching and the number of itches. HK did not produce any significant inhibition in histamine induced contraction in guinea pig ileum. From the above findings we conclude that the HK possesses antiallergic activity mediated by reducing of the release mediators from mast cells and also by 5-HT antagonism without the involvement of histamine (H1) receptors.

Purpose: The aim of this work was to formulate sodium alginate nanospheres of amphotericin B by controlled gellification method and to evaluate the role of the nanospheres as a “passive carrier” in targeted antifungal therapy. Methods: Sodium alginate nanospheres of amphotericin B were prepared by controlled gellification method, and the particle size analysis was carried out by scanning electron microscopy. The carrier capacity of sodium alginate was evaluated in terms of drug to polymer ratio. In vitro release study was carried out on all drug loaded nanospheres by the dialysis method. Release kinetics of drug from different drug loaded nanospheres was also determined. The in vivo antifungal efficacy of nanospheres bound drug vis-à-vis the free drug was evaluated in candidiasis- induced mice models. Results: Preparation of nanospheres through controlled gellification method yielded particles with a size range of 419.6 ± 0.28 nm. Studies on drug to polymer ratio showed a linear relationship between concentration of drug and drug loading capacity. In vitro release kinetic study revealed that the release of drug from the nanospheres followed Fickian diffusion. In vivo studies showed that the nanosphere-bound drug produced a higher antifungal efficacy than the free drug. Conclusion: The formulated sodium alginate nanospheres containing amphotericin B was found to have better antifungal activity when compared to the free drug and also yielded sustained in vitro release.