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Microsoft word - revfinalhormome…lc-1 tiff-1.doc
Deepti Varma & Dr. Susan Jansen, Temple University,
Philadelphia, PA and David J. Tognarelli, Jasco Inc., Easton, MD
Rapid Profiling of
Human Sex Hormones
Rapid separation of human sex
hormones in Biological Samples using
the JASCO X-LC system
Hypertension is a serious problem among African
breast cancer risk 3. Testosterone is a steroid hormone
American population, especially women.
from the androgen group which is present at much
According to the American Heart Association
higher concentration in men than in women.
(AHA), around 42.6% of African American menabove the age 20 and 46.6% of African Americanwomen have hypertension1. According to another
Figure 1: Conversion of Progesterone to Estrogenic
study conducted by AHA, hypertension was the
cause of death of 5,762 African American menand 6,664 African American women in 2004.1Men
and postmenopausal women are known to havegreater incidences of hypertension compared to
premenopausal women. This suggests that thefemale sex hormones have an effect on bloodpressure2. Estrone (E1), estradiol (E2), estriol (E3)
and progesterone are the main female sexhormones in the body. Out of all these, estrone is
assumed to be the only one present in variablequantities in postmenopausal women. 2-
hydroxyesterone (2-OHE1), 16_-hydroxyestrone(16_ -OHE1) and 4- steroid hormone from the
androgen group which is present at much higherconcentration in men than in women.
Determining the link between sex hormones withthe pathogenesis of hypertension is complex.
This application note describes a 6.1 min methodfor identifying a mixture of 8 sex hormones using
Figure 2: Metabolism of Estrogenic Compounds
Jasco X-LC. Separation of these compounds using
conventional HPLC typically takes around 16
minutes. hydroxyestrone (4-OHE1) are the majormetabolites of estrone. The ratio of 2-
hydroxyesterone (2-OHE1) to 16_-hydroxyestrone
(16_ -OHE1) is considered to be an index ofbreast cancer risk 3. HPLC typically takes around
16 minutes. hydroxyestrone (4-OHE1) are the
major metabolites of estrone. The ratio of 2-hydroxyesterone (2-OHE1) to 16_-hydroxyestrone(16_ -OHE1) is considered to be an index of
Results and Discussion
Figure 2 is an X-LC chromatogram of a standard
All the hormones were dissolved in 1 ml methanol
mixture of hormones (50 ng each). The compounds
such that the stock mass concentration for all was
are separated by a gradient elution of water and
1mg/ml.Working standards for the analytes were
acetonitrile on a Restek Pinnacle DB Biphenyl 1.9_m
prepared by serial dilution using ACN. 2-methoxy
(50 x 2.1 mm) at a flow rate of 0.165 ml/min.
Estrone, 4-hydroxy estrone were obtained from
Wavelength 220 nm was used for the detection of
these compounds. Resolution of testosterone from
hydroxyestrone were obtained from Steraloids,
estrone, is 1.25 in this method, acceptable for
estrone, E1; estradiol,E2; estriol, E3; were
quantitative determinations of both species.
X-LC chromatogram of a standard
XLC system and operating conditions:
mixture of hormones.(critical pair
Mobile Phase A: WaterMobile Phase B: ACNFlow Rate: 0.165 ml/min
1. American Heart Association- Statistical Fact S
2. Khalil AR, Hypertension, 2005, 46, 249-254.
3. Ursin G et al, Environ Health Perspect, 1997, 105,
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