Supported liquid extraction in combination with LC-MS/MS for high-throughput quantitative analysis of hydrocortisone in mouse serum.
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.
Department of Drug Metabolism and Pharmacokinetics, Novartis Institutes for Biomedical Research, One Health Plaza, East Hanover, NJ 07936, USA.
Shari
S
Wu
Wenkui
W
Li
Tapan
T
Mujamdar
Tom
T
Smith
Matthew
M
Bryant
Francis L S
FL
Tse
D000818:Animals; D001769:Blood; D005591:Chemical Fractionation / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006854:Hydrocortisone / Q000097:blood*; D051379:Mice / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D006854:Hydrocortisone