Determination of 4-Chloro-2-chloromethyl-5-(4-chlorophenyl)thieno[2,3-d]- pyrimidine in Rabbit Serum by HPLC and its Application in a Pharmacokinetic Study

Abstract:

In drug development, the analytical method development including the bioanalytical method development is essential and useful for the pharmacokinetic study of the lead molecule and is an important step. A sensitive method for the quantification of potential anti lipid lowering 4-chloro-2-chloromethyl-5-(4- chlorophenyl)thieno[2,3-d]pyrimidine 5 (SCOP/2007/1) in rabbit serum using high-performance liquid chromatography (HPLC) was developed. Sample pretreatment involved a simple protein precipitation by the addition of 1.5 ml of acetonitrile to 0.5 ml serum sample volume. Separation was achieved on Agilent- C18 (5μm,150 mm×4.6mm) reversed-phase column at 40°C with acetonitrile: water (80:20, v/v) at a flow rate of 0.6 ml/min. The calibration curve of 5 in serum showed good linearity over the concentration range of 25–6400 ng/ml. The limit of detection and limit of quantification were 7.81 ng/ml and 23.69 ng/ml, respectively. Intra and inter-day precisions in all samples were within 15%. The validated method was successfully applied to a preclinical pharmacokinetic study of 5 in rabbits. After oral administration of 100 mg/kg 5 to rabbits, the main pharmacokinetic parameters T_max , C _max , T_1/2 Kel, Ka and AUC were 2 h, 1297.28 ng/ml-1, , 0–t 0.495 h,1.4 h-1, 3.99 h-1 and 2930.5 ng h-1 L-1, respectively.

Keywords: Antihyperlipideamic; SCOP/2007/1; Bioanalytical; Pharmacokinetics; Rabbit Serum; Oral administration