Open AccessRobotic inhibition assay for determination of HMG‐CoA reductase inhibitors in human plasma
Author(s) -
Fang Wei,
Liu Lida,
Hsieh John Y.K.,
Zhao Jamie,
Matuszewski Bogdan K.,
Rogers John D.,
Dobrinska Michael R.
Publication year - 2002
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.10046
Subject(s) - simvastatin , chemistry , reproducibility , chromatography , reductase , coefficient of variation , enzyme , pharmacology , biochemistry , medicine
The cholesterol‐lowering drug simvastatin (SIMV, Zocor ® ) reduced heart attacks by 42% in patients who had high cholesterol levels and suffered from heart disease. Upon oral administration, SIMV is quickly hydrolyzed to its β‐hydroxyacid and other acid metabolites, which are potent inhibitors of 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase. A Tecan‐based enzyme inhibition assay has been developed to improve the existing Zymark‐based assay for the determination of both active and total concentrations of HMG‐CoA reductase inhibitors in human plasma. A Tecan Genesis 200 robotic workstation equipped with eight probes and customized hardware was utilized to achieve higher sample throughput and improve assay reproducibility and mechanical stability. The developed enzyme inhibition assay was validated over two concentration ranges of 0.4–20 ng equivalent/mL, and 2–50 ng equivalent/mL. Intra‐ and interday precision data (coefficient of variation (CV)) for both concentration ranges were less than 9%, with an accuracy of 93–107%. The interday precision for the determination of quality control (QC) samples was less than 2% and 8%, respectively. The respective interday QC accuracy values were 93–103% and 97–104%. Good linearity across the two concentration ranges was observed, with acceptable reproducibility. This improved enzyme inhibition assay has been utilized to analyze human plasma samples from several clinical studies. J. Clin. Lab. Anal. 16:209–215, 2002. © 2002 Wiley‐Liss, Inc.