Guoxiu Wei, Susan A Jansen, Sheara Williamson and Michael Brown
Aim: Prostanoids and female sex hormones play an important role in the progression of hypertension. However, their close-related structures make simultaneous separation and analyses difficult. A method was developed to separate and analyze seventeen important bioactive compounds including nine prostanoids and eight sex hormones in a short time. By applying the method to quantify prostanoids and female sex hormones in urine samples from hypertension patients, it becomes possible to find effective biomarkers for clinical diagnosis, prevention and treatment of hypertension.
Materials and methods: A reversed phase high performance liquid chromatography is used to develop the separation method. A combination of up to nine prostanoids (PGD2, PGE2, 6-keto PGF1α, PGF2α, 11-dehydro TXB2, 8-iso PGF2α, 13,14-dihydro-15-keto PGA2, 13,14-dihydro-15-keto PGE2 and 15-deoxy Δ12,14 PGJ2) and eight female sex hormones (E1, E2, E3, progesterone, 2-OHE1, 4-OHE1, 16α-OHE1, and 2-MeOE1) are chosen as analytes. The separation is performed on Symmetry C18 4.6x250 mm column with 5 μm particle size. A UV detector is selected to determine the levels of analytes in a single 25-minute run.
Results: The method is the first we are aware of successfully demonstrating the separation of the two different groups of bioactive compounds prostanoids and female sex hormones. It has been shown to have excellent selectivity, sensitivity, and accuracy over biologically relevant concentration ranges and has been tested on urine samples from hypertension patients.
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