Year: 2020 | Month: June | Volume 10 | Issue 3
Estimation of In-vitro Plasma Protein Binding of Ampicillin in Horses Based on Spectrophotometric Method
Kondampati Kasturi Devi
Milindmitra Lonare
Saloni Singla
Suresh Kumar Sharma
Vinod Kumar Dumka
Simrat Pal Singh Saini
DOI:10.30954/2277-940X.03.2020.4
Abstract:
Estimation of plasma protein binding (PPB) is of paramount importance in the pharmacokinetics characterization of drugs, as it can cause significant change in volume of distribution, clearance and half-life of the drug. Ampicillin (α-amino benzyl penicillin) is most commonly used drug in equine practice. This study was conducted to determine the extent of PPB of ampicillin in apparently healthy horses (n=6). A simple spectrophotometric method was applied for the determination of ampicillin at 320 nm wavelength, based on acid degradation product of penicillin at 75°C in presence of citrate buffer (pH 5.2) and traces of copper salt. In the study, it was observed that this method permits the detection of ampicillin to a level not beyond 1.0 μg/ml. Various concentrations of ampicillin (3.125, 6.25, 12.5, 25, 50, 100 μg/ml) were prepared in triplicate in pooled plasma collected from healthy animals. In vitro binding of ampicillin to plasma proteins was determined by employing the equilibrium dialysis technique. The study revealed that the plasma protein binding of ampicillin was to the extent of 12.8 ± 0.07 %. Binding capacity of ampicillin to plasma protein (βi) and dissociation rate constant of protein-drug complex (Kβ) in the present study were 0.34 × 10-6 ± 0.02 × 10-6 mol.gm-1 and 0.003 × 10-9 ± 0.0003 × 10-9 mol, respectively in horses. Hence, the study concluded that usage of spectrophotometric method helps in quick, cost effective and efficient results in estimation of PPB for ampicillin.
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