title: Sirolimus quantification by high-performance liquid chromatography with ultraviolet detection.
abstract: The need to adapt optimal conditions of sirolimus blood level monitoring in laboratories led us to optimize an high-performance liquid chromatography-ultraviolet method and compare the elution performances using the mobile phase A, 68% MeOH/2% acetonitrile (ACN)/30% H(2)O and mobile phase B, 30% MeOH/42% ACN/28% H(2)O. Samples were assayed with 1-chlorobutane, redissolved in MeOH/water and injected onto a C-18 column at 50 degrees C. The assay achieved sensitivity of 2.5-150 ng/ml (CV = 10.6%) and recovery of 92-103.6%. The intra- and interassay precisions ranged from 3.3% to 13% and from 5.9% to 15% for quality controls of 7.5, 60 and 120 ng/ml. The mobile phase A was unable to elute and recover sirolimus and internal standard in the expected retention time and concentration. Under our working conditions, the assay was precise, accurate and sensible, stressing the importance of establishing for the best working conditions according to the staff and demands of the laboratory.
authors.affiliation: Department of Medicine-Nephrology Division, Federal University of São Paulo, Escola Paulista de Medecina, Rua Botucatu 740, 5574-6300 São Paulo, Brazil.
authors.forename: Maria Claudina Camargo de
authors.initials: MC
authors.lastname: Andrade
authors.forename: Giovana Seno
authors.initials: GS
authors.lastname: Di Marco
authors.forename: Claudia Rosso
authors.initials: CR
authors.lastname: Felipe
authors.forename: Fernando
authors.initials: F
authors.lastname: Alfieri
authors.forename: Hélio Tedesco Silva
authors.initials: HT
authors.lastname: Júnior
authors.forename: José Osmar Medina
authors.initials: JO
authors.lastname: Pestana
authors.forename: Dulce Elena
authors.initials: DE
authors.lastname: Casarini
mesh_terms: D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007166:Immunosuppressive Agents / Q000097:blood*; D020123:Sirolimus / Q000097:blood*; D014466:Ultraviolet Rays
chemical_list: D007166:Immunosuppressive Agents; D020123:Sirolimus