Post‐chromatographic Detection of Progesterone Using Terbium‐Sensitized Luminescence

ABSTRACT

This report deals with a time‐resolved fluorometric determination of progesterone by reversed‐phase high‐performance liquid chromatography. The method was simple to use and performed on a C18 column, with acetonitrile/water 80:20 as mobile phase (flushed at 1 mL · min⁻¹), and a 0.03 mol · L⁻¹ Tb(NO₃)₃, 0.005 mol · L⁻¹ SDS methanolic miscellaneous solution as post‐column reagent (added at 0.8 mL · min⁻¹). The validation of the assay was performed for a 200‐µL injection, and it showed that this method presented no interferences with estrogens, and that it was linear in the range 5 × 10⁻⁶–75 × 10⁻⁶ mol · L⁻¹ (correlation coefficient of 0.99), repeatable (3.7% intra‐day relative standard deviation (RSD) and 5.6% inter‐day RSD, at 40 × 10⁻⁶ mol · L⁻¹), with a detection limit at 2.5 × 10⁻⁶ mol · L⁻¹, and a quantification limit at 5 × 10⁻⁶ mol · L⁻¹.

Keywords:

• High‐performance liquid chromatography
• Luminescence
• Progesterone
• Terbium
• Complexes

ACKNOWLEDGEMENT

The authors are grateful to Mrs. Pierre‐Eugène for technical assistance.