Thalidomide enantiomers: determination in biological samples by HPLC and vancomycin-CSP.
Thalidomide is a racemate with potentially different pharmacokinetics and pharmacodynamics of the component (+)-(R)- and (-)-(S)-thalidomide enantiomers. As part of a project on the adjunctive effects of thalidomide and cytotoxic agents, a method for the chiral separation and quantitation of thalidomide was developed and validated. Thalidomide in relevant serum and tissue homogenate samples was stabilized by buffering with an equal volume of citrate-phosphate buffer (pH 2, 0.2M), and stored at -80 degrees C pending assay. The thalidomide enantiomers, extracted from the samples with diethyl ether, were well separated on a chiral HPLC column of vancomycin stationary phase and a mobile phase of 14% acetonitrile in 20 mM ammonium formate adjusted to pH 5.4; their concentrations were determined with phenacetin as internal standard at 220 nm detection. Over a thalidomide concentration range of 0.1-20 microg/ml, assay precision was 1-5% (CV) for both enantiomers, and calibration curves were linear with all correlation coefficients being >0.99. The estimated limit of quantification for both enantiomers was 0.05 microg/ml with 0.2-0.6 ml serum samples. Thalidomide in rat and human serum, acidified and stored as described above, was found to be chemically and chirally stable over 1 year. The method has been successfully applied to serum samples from human patients undergoing thalidomide treatment for mesothelioma, and to serum, blood and tissue samples from a laboratory rodent model using transplanted 9l gliosarcoma. Enantioselectivity in thalidomide pharmacokinetics has been found, thereby reinforcing the need for considering the relevance of chirality in thalidomide pharmacology.
Murphy-Poulton
Susan F
SF
Bill Walsh Cancer Research Laboratories, Department of Medical Oncology, University of Sydney at Royal North Shore Hospital, St. Leonards, N.S.W. 2065, Australia.
Boyle
Frances
F
Gu
Xiao Qing
XQ
Mather
Laurence E
LE
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods*; D004355:Drug Stability; D005260:Female; D006801:Humans; D051381:Rats; D011916:Rats, Inbred F344; D015203:Reproducibility of Results; D013237:Stereoisomerism; D013792:Thalidomide / Q000032:analysis* / Q000302:isolation & purification / Q000493:pharmacokinetics; D014640:Vancomycin / Q000737:chemistry*
D013792:Thalidomide; D014640:Vancomycin