6^th World Congress on Bioavailability & Bioequivalence: BA/BE Studies Summit August 17-19, 2015 Chicago, USA

S Raghunadha Reddy has completed his PhD from Jawaharlal Nehru Technological University, Anantapur and currently doing Postdoctoral studies from Department of Pharmaceutical Science, School of Pharmacy, University of Maryland. Previously he was worked as Head of Quality Assurance and Regulatory Affairs at Clinsync Clinical Research Pvt. Ltd. He has published 17 papers in reputed journals and has been serving as an Editorial Board Member of Journal of Comprehencive Pharmacy. He has extensive experience in Good Clinical Practice-ICH, Good Laboratory Practice, QMS (ISO9001-2008), Bioanalytical method Development and validation, Computer System Validations (21 CFR Part-11) and Regulatory Affairs.

An open-labeled, balanced, single-dose, two treatment, two period, two sequence, randomized two way crossover study was conducted in 18 healthy adult male subjects to determine the pharmacokinetic, bioavailability and bioequivalence of carvedilol phosphate 40 mg extended release capsules in comparison with Coreg CRTM extended release capsules after single dose administration under fed conditions with a wash-out period of at least 7 days was used. Each volunteer received a 40 mg capsule of the reference (or) test drug, respectively. On the day of dosing, blood samples were collected before dosing (at 0.0 hr) and 0.50, 1.00, 1.50, 2.00, 2.50, 3.00, 3.50, 4.00, 4.50, 5.00, 5.50, 6.00, 6.50, 7.00, 7.50, 8.00, 10.00, 12.00, 16.00, 24.00, 36.00 and 48.00 hours after dosing. Analysis of carvedilol and its metabolite 4-hydroxy phenyl carvedilol concentrations was performed using a validated LC-MS/MS method. The pharmacokinetic parameters were analyzed using the non-compartmental model. Drug safety and tolerability were assessed. The primary pharmacokinetic parameters at 90% CI were within the 80 to 125% interval required for bioequivalence as stipulated in the current regulations of the USFDA acceptance criteria. The geometric mean ratios (Test/Reference) between the two products of extended-release carvedilol capsule under fed condition were 114.41% (93.68%-116.74%) and 113.15% (96.67%-122.45%) for Cmax ratios, 101.54% (95.73-104.85%) and 102.72%(95.12%-113.35%) for AUC0-t ratios and 104.56% (103.24%-107.58%) and 105.73% (95.45%-110.50%) for AUC0-inf ratios of carvedilol and its metabolite 4-hydroxy phenyl carvedilol respectively. 18 volunteers had completed both treatments. There was no significant difference of the Tmax parameter between the two formulations (p>0.05). No serious adverse events related to the study drugs were found. This single dose study found that the test formulation carvedilol phosphate ER capsules is bioequivalent in terms of rate and extent of absorption to the reference formulation Coreg CRTM ER capsules of 40 mg under fed condition in healthy adult male volunteers according to the USFDA regulatory guidance.

S Raghunadha Reddy has completed his PhD from Jawaharlal Nehru Technological University, Anantapur and currently doing Postdoctoral studies from Department of Pharmaceutical Science, School of Pharmacy, University of Maryland. Previously, he was worked as Head of Quality Assurance and Regulatory Affairs at Clinsync Clinical Research Pvt. Ltd. He has published 17 papers in reputed journals and has been serving as an Editorial Board Member of Journal of Comprehencive Pharmacy. He has extensive experience in Good Clinical Practice-ICH, Good Laboratory Practice, QMS (ISO9001-2008), Bioanalytical method Development and validation, Computer System Validations (21 CFR Part-11) and Regulatory Affairs.

A fast and highly sensitive ultra-high performance liquid chromatography (UPLC) method for the determination of phenelzine in human plasma have been developed using tandem mass spectrometry (MS/MS) detection. Hydroxyzine was used as an internal standard(IS). The extraction of the phenelzine from human plasma was performed using solid phase extraction. ACE-C18 (5µm, 100x4.6 mm) reverse phase column was employed for chromatographic separation of analyte and internal standard for MS/MS detection at 0.9 ml/min flow. Detection was performed at transitions of m/z 137.258→106.906 for phenelzine and m/z 376.300→ 202.100 for hydroxyzine by positive electro-spray ionization (ESI+) in multiple reaction monitoring (MRM) mode using tandem mass spectrometry. The developed method was compared in the terms of validation parameters including linearity, sensitivity, precision and accuracy. The analysis was carried out in 3.5 min and the matrix matched calibration curves in the range of 0.508 ng/mL to 25.144 ng/mL were used for quantification with the correlation coefficients demonstrating good linearity (0.996-0.999). The mean extraction recoveries for phenelzine and IS from plasma were 96.5% and 88.3%, respectively. Matrix based samples were stable at room temperature for 12 hrs, processed samples were stable at least for 28 hrs and also stable at six freeze-thaw cycles. This method was successfully applied for determination of phenelzine in human plasma for pharmacokinetic study.

S Raghunadha Reddy has completed his PhD from Jawaharlal Nehru Technological University, Anantapur and currently doing Postdoctoral studies from Department of Pharmaceutical Science, School of Pharmacy, University of Maryland. Previously he was worked as Head of Quality Assurance and Regulatory Affairs at Clinsync Clinical Research Pvt. Ltd. He has published 17 papers in reputed journals and has been serving as an Editorial Board Member of Journal of Comprehencive Pharmacy. He has extensive experience in Good Clinical Practice-ICH, Good Laboratory Practice, QMS (ISO9001-2008), Bioanalytical method Development and validation, Computer System Validations (21 CFR Part-11) and Regulatory Affairs.

A simple, sensitive and reliable method have been developed for simultaneous quantification of Hydroxyzine and Cetirizine in human plasma by using on-line robotic HTLC–MS/MS method. Quetiapine was used as an internal standard(IS). A typical two-column setup featuring two six-port switching valves was employed for method development and Turbo flow on-line technique was applied for separation of analyte from plasma sample. The analytical procedure involves on-line coupling of sample extraction with Cyclone P (50 mm × 0.5 mm 50 µm) HTLC column by injecting 15μL sample and chromatographic separation is performed with Inertsil ODS-3(5 µm ,4.6 x100 mm.). To achieve required chromatograms with consistency we have performed different combinations of the solvents and gradient system. Finally we succeeded with the solution combinations of 5mM Ammonium Acetate:Methanol: Acetonitrile (5:5:90) in pump A, Pure methanol in pump B, 0.1% formic acid in pump C and washing solution in the ratio of 70:25:5 (methanol:water:IPA) in pump D and analyzed more than 150 samples with out overloading of the chromatographic columns with improved real throughput efficiency. Detection was performed at transitions of m/z 375.300→201.100 for Hydroxyzine, m/z 389.200→201.100 for Cetirizine and m/z 384.200→253.100 for Quetiapine by positive electro-spray ionization (ESI+) in multiple reaction monitoring (MRM) mode using tandem mass spectrometry. The calibration curves were linear over a concentration range of 0.100 ng/mL to 150.000 ng/mL of Hydroxyzine and 0.500 ng/mL to 500.000 ng/mL of Cetirizine for quantification with the correlation coefficients demonstrating good linearity (0.994-0.999). The total run time of analysis was 5 min and the lower limits of Quantification were 0.100 ng/mL for Hydroxyzine and 0.500 ng/mL for Cetirizine, respectively. The method validation was carried out in terms of specificity, sensitivity, linearity, precision, accuracy and stability. The validated method was successfully applied in bioavailability and bioequivalence study.