Reversed-phase high-performance liquid chromatographic determination of 7 alpha-hydroxy-4-cholesten-3-one in human serum.
A method for the determination of 7 alpha-hydroxy-4-cholesten-3-one in human serum has been developed. The method is based upon solid-phase (C18) extraction of serum after addition of 7-beta-hydroxy-4-cholesten-3-one as internal standard. The extract is analysed by reversed-phase HPLC using acetonitrile-water as a mobile phase. The effluent is monitored at 241 nm. The overall recoveries of the method range from 96% to 105%. The coefficient of variation for the within-run precision is 3.2% (n = 20, mean = 13.0 ng/ml) and for the between-run precision 3.8% (n = 32, mean = 13.3 ng/ml). The limit of detection is 3 ng at a signal-to-noise ratio of 3:1, which corresponds to 1 ng/ml using 3 ml of serum. The median value of 7 alpha-hydroxy-4-cholesten-3-one found in blood donors (n = 27) was 8.9 ng/ml (range 2-35 ng/ml).
Department of Clinical Chemistry, Mälarsjukhuset, Eskilstuna, Sweden.
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Pettersson
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Eriksson
D000097:Acetonitriles; D000328:Adult; D002783:Cholestenones / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D008875:Middle Aged; D011786:Quality Control; D012016:Reference Values; D012680:Sensitivity and Specificity; D014867:Water
D000097:Acetonitriles; D002783:Cholestenones; D014867:Water; C002656:7 alpha-hydroxy-4-cholesten-3-one; C032159:acetonitrile