Validated enzyme-linked immunosorbent assay for determination of rosuvastatin in plasma at picogram level.
Drug Test Anal
; 5(5): 334-9, 2013 May.
Article
em En
| MEDLINE
| ID: mdl-21998019
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Pirimidinas
/
Sulfonamidas
/
Inibidores de Hidroximetilglutaril-CoA Redutases
/
Fluorbenzenos
Limite:
Animals
Idioma:
En
Revista:
Drug Test Anal
Assunto da revista:
FARMACOLOGIA
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
Arábia Saudita