Quantitation of promethazine enantiomers in human serum using a chiralcel OJ-R column and mixed-mode disc solid-phase extraction.
A liquid chromatographic method was developed for the assay of R(+)- and S(-)-promethazine enantiomers from human serum. The method involves the use of the mixed mode disc solid-phase extraction technique for sample clean-up. Chromatographic resolution of the enantiomers was performed on a reversed-phase cellulose-based chiral column (Chiralcel OJ-R) under isocratic conditions using a mobile phase consisting of 0.5 M aqueous sodium perchlorate/acetonitrile (63:37, v/v) at a flow rate of 0.5 ml min-1. Recoveries in the range of 97-99% at 20 ng ml-1 levels were obtained for both promethazine enantiomers. Intra-day and inter-day precision calculated as R.S.D.% was in the 3-8% ranges for both enantiomers. Intra-day and inter-day accuracy calculated as percent error was in the 0-10 and 1-7% ranges for both enantiomers, respectively. Linear calibration curves were obtained for each enantiomer in serum in the concentration range 5-90 ng ml-1. The limit of quantitation of each enantiomer was 10 ng ml-1. The detection limit for each enantiomer in serum using UV detection at 249 nm was 2 ng ml-1 (S/N = 2).
Liu
J
J
Department of Medicinal Chemistry, College of Pharmacy, University of Georgia, Athens 30602-2352, USA.
Stewart
J T
JT
D018926:Anti-Allergic Agents / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D000889:Anti-Arrhythmia Agents / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D011398:Promethazine / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D014700:Verapamil / Q000737:chemistry
D018926:Anti-Allergic Agents; D000889:Anti-Arrhythmia Agents; D014700:Verapamil; D011398:Promethazine