Use of cyclodextrins for prepurification of progesterone and testosterone from human serum prior to determination with isotope dilution gas chromatography/mass spectrometry.
A new application of cyclodextrins is presented, being their use for sample purification. This new application is exemplified with a method for the determination of progesterone and testosterone in human serum based on isotope dilution gas chromatography/mass spectrometry. The whole procedure consists of a traditional solvent extraction with hexane for progesterone, respectively dichloromethane for testosterone, followed by a back-extraction of the steroids into a cyclodextrin-water solution. After washing of the aqueous phase with hexane, the steroids are finally extracted with toluene (progesterone) and dichloromethane (testosterone). Different cyclodextrins and cyclodextrin derivatives in various concentrations are investigated. A solution of 150 mmol/L hydroxypropyl beta-cyclodextrin in water is selected. The recovery of progesterone and testosterone is 70 and 80% respectively. The purification effectiveness, accuracy, and precision of the new method are equivalent to a traditional method using gel chromatography for sample purification.
Thienpont
L M
LM
Laboratoria voor Medische Biochemie en Klinische Analyse, University of Ghent, Belgium.
De Brabandere
V I
VI
Stöckl
D
D
De Leenheer
A P
AP
D003505:Cyclodextrins / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry; D006586:Hexanes; D006801:Humans; D007553:Isotope Labeling; D008752:Methylene Chloride; D011374:Progesterone / Q000097:blood* / Q000302:isolation & purification; D013739:Testosterone / Q000097:blood* / Q000302:isolation & purification
D003505:Cyclodextrins; D006586:Hexanes; D013739:Testosterone; D011374:Progesterone; D008752:Methylene Chloride