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A new method for simultaneous determination of catecholamines in reproductive organs from rats by high performance liquid chromatography with electrochemical detection

The aim of this research was to propose a new sensitive and fast method to the simultaneous determination of catecholamines (noradrenaline and adrenaline) in samples of reproductive organs from male rats by High Performance Liquid Chromatography with Electrochemical Detection. The method involved direct injection of acid extract on to a serum albumin dimethylocadecyl-silane (HSAC18) and the utilization of 0.013mol L-1 methanosulfonic acid (pH = 3.0): acetonitrile (96:4v/v), 0.033g heptanosulfonic acid and 0.01g EDTA as mobile phase. The detection was obtained using an electrochemical detector L-ECD-6A-Shimadzu with a potential of the 85mV. percloric acid 0.4 mol L-1 was used in the procedure of catecholamine extraction. The extraction of the analyte resulted in recovery values between 56% and 114%, and coefficient of variation between 3.8% and 23%. The quantification limit of the method was between 0.008 and 0.030ng mg-1 for noradrenaline, and 0.009 and 0.051ng mg-1 for adrenaline. In conclusion, the analytical method was validated and applied to the real samples of rats, allowing the determination of low concentrations of the catecholamines studied. The method has various advantages: fast, high precision and good selectivity and do not require sample treatment.

catecholamines; reproductive organs; column ISRP- C18; HPLC-ECD


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