A liquid chromatography-mass spectrometry method for the quantitative analysis of urinary endogenous estrogen metabolites

Nat Protoc. 2007;2(6):1350-5. doi: 10.1038/nprot.2007.176.


The ability to measure estrogen metabolites (EMs) quantitatively is important for investigating their individual roles in cancer screening, treatment and prevention, as well as in a host of other hormone-related disorders. In this protocol we describe a method that is capable of quantitating 15 distinct EMs in urine. Endogenous EMs are quantitatively measured using a liquid chromatography-tandem mass spectrometry method in which the spectrometer operates in a selected reaction monitoring mode. This method is capable of quantifying estrone and its 2-, and 4- and 16alpha-hydroxy and its 2-, 4-methoxy derivatives, and 2-hydroxyestrone-3-methyl ether; 17beta-estradiol and its 2-hydroxy, and 2- and 4-methoxy derivates, and estriol, 16-epiestriol, 17-epiestriol, and 16-ketoestradiol. The method requires only 0.5 ml of urine and approximately 60 urine samples can be quantitatively analyzed per week.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural

MeSH terms

  • Animals
  • Chromatography, High Pressure Liquid / methods*
  • Estrogens / chemistry
  • Estrogens / metabolism*
  • Estrogens / urine*
  • Female
  • Humans
  • Mass Spectrometry / methods*
  • Molecular Structure
  • Reproducibility of Results
  • Sensitivity and Specificity


  • Estrogens