title: Simultaneous determination of tramadol and O-desmethyltramadol in human plasma using HPLC-DAD.
abstract: A sensitive and accurate method for the extraction and quantification of tramadol (T) and its active metabolite, O-desmethyltramadol (ODT) in human plasma with high-performance liquid chromatography-diode array detection was developed and validated. The analytes were extracted from plasma samples by tert-butylmethyl ether in the presence of ammonium hydroxide as alkaline medium and back extraction with 1.0 M hydrochloric acid. Propranolol was used as internal standard. The extraction efficiencies of T and ODT were 83.51 and 78.72%, respectively. The calibration curves were linear (r(2) > 0.99) in the concentration range of 250-2000 ng/mL for T and ODT. Limits of detection and quantification were 125 and 250 ng/mL for both analytes. Intra- and interassay precision for T and ODT were ranged from 1.89 to 10.91% and 2.16 to 5.85%, respectively. Intra- and interassay accuracy for T and ODT were ranged from -13.07 to 4.99% and -2.03 to -6.98%, respectively. The method was successfully applied to quantify T and ODT from authentic plasma samples received from Hospital Sohag University. The method was completely validated and can be of interest to clinical and forensic laboratories.
authors.affiliation: Forensic Medicine and Clinical Toxicology Department, Faculty of Medicine, Sohag University, Sohag, Egypt.
authors.forename: Maha A
authors.initials: MA
authors.lastname: Hilal
authors.affiliation: Assiut Chemical Laboratory, Medico-Legal Department, Ministry of Justice, Justice, Egypt khaled.masoud@yahoo.com.
authors.forename: Khaled M
authors.initials: KM
authors.lastname: Mohamed
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D002021:Buffers; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D010710:Phosphates / Q000737:chemistry; D017680:Potassium Compounds / Q000737:chemistry; D011433:Propranolol / Q000097:blood; D015203:Reproducibility of Results; D014147:Tramadol / Q000031:analogs & derivatives* / Q000097:blood*
chemical_list: D000097:Acetonitriles; D002021:Buffers; D010710:Phosphates; D017680:Potassium Compounds; C080580:O-demethyltramadol; D014147:Tramadol; D011433:Propranolol; C013216:potassium phosphate; C032159:acetonitrile