THE ANALYSIS OF TRAMADOL AND ITS MAJOR METABOLITE O-DESMETHYL
Unit 2, Kelvin Campus, West of Scotland Science Park, Glasgow. G20 0SP
TRAMADOL, FROM HUMAN PLASMA USING POSITIVE TURBOIONSPRAY LC-MS/MS Lindsay Best, Caroline Wood, Jim Hillier I N T R O D U C T I O N
Tramadol is an analgesic used to treat moderate to moderately severe pain. A specific analytical
Back Calculated Concentrations from the Calibration Curve for Tramadol
method has been established and validated for the determination of tramadol and its major
Figure 1: Representative Calibration Figure 2: Representative Calibration
metabolite o-desmethyl tramadol in human plasma. The method was validated over the range 5.00
Curve for Tramadol Concentration of Tramadol (ng/ml) Curve for O-Desmethyl Tramadol M E T H O D O L O G Y Sample Preparation and Extraction - Plasma Samples
Control human plasma samples were fortified with solutions containing tramadol and o-desmethyltramadol. These plasma samples were then extracted by liquid/liquid extraction using methyl tert
butyl ether (MTBE). The resultant supernatant was evaporated to dryness followed by reconstitutionin mobile phase. Back Calculated Concentrations from the Calibration Curve for O-Desmethyl Tramadol Sample Analysis Concentration of O-Desmethyl Tramadol (ng/ml) METHODOLOGY
Applied Biosystems MDS SCIEX API 3000 TandemMass Spectrometer with the TurboIonSprayTM Source
Agilent HP1100 Series Quaternary Pump andDegasser.
20mM Ammonium Acetate/Acetonitrile (50/50; v/v)
Intra-Occasion Statistics for Tramadol Figure 3: Representative Chromatogram from a Calibration Standard Sample containing Concentration of Tramadol (ng/ml) 5.00ng/ml of Tramadol and O-Desmethyl Tramadol V A L I D A T I O N R E S U L T S
Back-calculated concentrations for the plasma calibration samples are shown in Tables 1 and 2.
The calibration curves showed good linearity over the range 1.00 - 500 ng/ml. Typical calibration
curves are displayed in Figure 1 and 2. Examples of chromatography from a calibration standardcontaining 1.00 and 500 ng/ml of each analyte are presented in Figures 3 and 4.
Intra-day statistics are presented in Tables 3 and 4.
The inter-day (three occasions) bias for tramadol was 0.8, 0.0, -0.4, -1.0 and -2.2% at 5.00, 12.5,
Intra-Occasion Statistics for O-Desmethyl Tramadol
50.0, 400 and 500ng/ml respectively. The inter-batch CV for tramadol at these concentrations was7.3, 4.7, 3.2, 2.8 and 4.0% respectively. The inter-batch (three occasions) bias for o-desmethyl
Concentration of O-Desmethyl Tramadol (ng/ml) Figure 4: Representative Chromatogram from a Calibration Standard Sample containing
tramadol was 3.8, 0.8, 2.2, -3.3 and -6.2% at 5.00, 12.5, 50.0, 400 and 500ng/ml respectively. The
500ng/ml of Tramadol and O-Desmethyl Tramadol
inter-batch CV for o-desmethyl tramadol at these concentrations was 6.9, 6.2, 5.5, 5.9 and 5.8%respectively. These results demonstrate good accuracy and precision over the range 1.00 to 500
Both analytes were also shown to be stable in plasma for 24hours at room temperature, over 3
freeze/thaw cycles and plasma extracts containing the analytes demonstrated stability for up to 72hours when stored at 4°C±3°C. C O N C L U S I O N
A sensitive and versatile bioanalytical LC-MS/MS method was developed and validated to
determine concentrations of tramadol and o-desmethyl tramadol in human plasma samples.
UMA PUBLICAÇÃO DA DYNAMO ADMINISTRAÇÃO DE RECURSOS LTDA. – 4º TRIMESTRE DE 2003 www.dynamo.com.br As cotas do Dynamo Cougar apre- IBX, o que não acontecia desde 1998. Aentre os pontos de análise para escolha deciaram-se 20,9% no quarto trimestre, atin- valorização do real frente ao dólar provo-nossos investimentos aqui na Dynamo. Trata-gindo uma valorização de 58,6% em 2003.
The Elmhurst SVCs Dynamic reactive power support for Northern Illinois In November of 2008 Siemens Energy, Inc. was awarded a turnkey contract for two (2) 138 kV Static Var Compensators (SVC’s) by Commonwealth Edison Company (ComEd) for the Elmhurst Substation, located a few miles south of the O’Hare International Airport in Chicago, Illinois. The installation of the Elmhurst SVCs