title: Validated enzyme-linked immunosorbent assay for determination of rosuvastatin in plasma at picogram level.
abstract: In this study, a highly sensitive enzyme-linked immunosorbent assay (ELISA) has been developed and validated for the determination of rosuvastatin (ROS) in plasma samples at picogram level. The assay employed a polyclonal antibody that specifically recognizes ROS with high affinity, and ROS conjugate of bovine serum albumin (ROS-BSA) immobilized onto microplate wells as a solid phase. The assay involved a competitive binding reaction between ROS, in plasma sample, and the immobilized ROS-BSA for the binding sites on a limited amount of the anti-ROS antibody. The bound anti-ROS antibody was quantified with horseradish peroxidase-labelled second anti-rabbit IgG antibody (HRP-IgG) and 3,3`,5,5`-tetramethylbenzidine (TMB) as a substrate for the peroxidase enzyme. The concentration of ROS in the sample was quantified by its ability to inhibit the binding of the anti-ROS antibody to the immobilized ROS-BSA and subsequently the colour intensity in the assay wells. The assay limit of detection was 25 pg ml(-1) and the effective working range at relative standard deviations (RSD) of ≤ 5% was 40-2000 pg ml(-1). Analytical recovery of ROS from spiked plasma was 96.2 - 104.8 ± 2.12 - 5.42%. The precision of the assay was satisfactory; RSD was 2.47 - 4.46 and 3.24 - 5.27% for the intra- and inter-assay precision, respectively. The analytical procedure is convenient, and one can analyze ~ 200 samples per working day, facilitating the processing of large-number batch of samples. The proposed ELISA has a great value in routine analysis of ROS for its pharmacokinetic studies.
authors: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Saudi Arabia. idarwish@ksu.edu.sa
authors: Ibrahim A
authors: IA
authors: Darwish
authors: Abdul-Rahman M
authors: AR
authors: Al-Obaid
authors: Hamoud A
authors: HA
authors: Al-Malaq
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis; D001667:Binding, Competitive; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005464:Fluorobenzenes / Q000097:blood* / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000032:analysis; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000097:blood* / Q000737:chemistry; D055767:Immobilized Proteins / Q000737:chemistry; D007074:Immunoglobulin G / Q000032:analysis; D057230:Limit of Detection; D011743:Pyrimidines / Q000097:blood* / Q000737:chemistry; D011817:Rabbits; D000068718:Rosuvastatin Calcium; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013449:Sulfonamides / Q000097:blood* / Q000737:chemistry
chemical_list: D000906:Antibodies; D005464:Fluorobenzenes; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; C051538:IGG-horseradish peroxidase; D055767:Immobilized Proteins; D007074:Immunoglobulin G; D011743:Pyrimidines; D013449:Sulfonamides; D012710:Serum Albumin, Bovine; D000068718:Rosuvastatin Calcium; D006735:Horseradish Peroxidase