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Separation of epimeric monohydroxylated metabolites of testosterone, androstenedione and thep-nitrobenzyl esters of hydroxylated 5β-cholanic acid using the same HPLC conditions

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Summary

The separation of authentic reference standards of epimeric monohydroxylated derivatives of both testosterone and lithocholic acidp-nitrobenzyl esters on a 10 μm Partisil-10 column with a single mobile phase of hexane-tetrahydrofuran-isopropanol (80∶15∶5) is reported. Baseline separation of all epimeric derivatives of the bile acid PNB-esters was achieved, and the principal testosterone metabolites (6β-, 7α-, 16β-, and 15β- and 16α-hydroxy-testosterones) were separated from one another and the parent compound. This system was used to separate metabolites of both androstene-dione and [14C]-lithocholic acid.

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Abbreviations

PNBDI:

O-p-nitrosobenzyl-N,N′-diisopropylisourea

PNB:

p-nitrobenzyl

HPLC:

High-performance liquid chromatography

THF:

Tetrahydrofuran

IPA:

Isopropanol

ITLC-SA:

Instant thin-layer chromatography — silic acid

S9:

9000 xg supernatant fraction

TLC:

Thin-layer chromatography

AUFS:

Absorbance units at full scale

tR :

Retention time

RRT:

Relative retention time

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Author information

Author notes

  1. J. C. Kawalek

    Present address: Division Veterinary Medicine, Food and Drug Administration, 20705, Beltsville, MD, USA

  2. K. -K. Hwang

    Present address: Microbiological Associates, Inc., 5221 River Rd., 20016, Bethesda, MD, USA

  3. M. I. Kelsey

    Present address: Division of Cancer Cause and Prevention, National Cancer Institute, Landow Building, 3C-37, Bethesda, MD, 20205, USA

Authors and Affiliations

  1. Chemical Carcinogenesis Program, Frederick Cancer Research Center, 21701, Frederick, MD, USA

    J. C. Kawalek, K. -K. Hwang & M. I. Kelsey

Authors
  1. J. C. Kawalek
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  2. K. -K. Hwang
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  3. M. I. Kelsey
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Kawalek, J.C., Hwang, K.K. & Kelsey, M.I. Separation of epimeric monohydroxylated metabolites of testosterone, androstenedione and thep-nitrobenzyl esters of hydroxylated 5β-cholanic acid using the same HPLC conditions. Chromatographia 14, 633–637 (1981). https://doi.org/10.1007/BF02291102

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  • DOI: https://doi.org/10.1007/BF02291102

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