Supported liquid extraction in combination with LC-MS/MS for high-throughput quantitative analysis of hydrocortisone...

A high-throughput LC-MS/MS bioanalytical method was developed and validated for the determination of hydrocortisone in mouse serum via supported liquid extraction (SLE) in a 96-well plate format. Although sample extracts from SLE result in similar matrix effects compared with conventional liquid-liquid extraction (LLE), greater analyte extraction recovery and much higher analysis throughput for the quantitative analysis of hydrocortisone in mouse serum were obtained. The current LC-MS/MS method was validated for a concentration range of 2.00-2000 ng/mL for hydrocortisone using a 0.100 mL volume of mouse serum. The intra- and inter-day precision and accuracy of the quality control samples at low, medium and high concentration levels showed < or =12.9% CV and -3.4-6.2% bias for the analyte in mouse serum.