<Originals>High-performance liquid chromatographic analysis of amoxapine and 8-hydroxyamoxapine in human serum

概要

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A method of isocratic liquid chromatographic determination of amoxapine and 8-hydroxyamoxapine, which are pharmacologically active antidepressant agents, was developed. A mixture of 20 mM potassium phosphate buffer (pH 3.0,containing 0.06 vol% n-nonylamine) and acetonitrile (85/15 vol%) is used as the mobile phase at a flow rate of 0.8 ml/min (ambient) and promazine is detected at a wavelength of 210 nm in comparison with the internal standard. As pretreatment, 2 N sodium hydroxide (0.1 ml) is added to the serum, followed by addition of, the internal standard. Extraction is performed with n-butyl chloride/ethyl acetate (1/1 vol%). The upper organic layer is mixed with 0.01 N hydrochloric acid (0.2 ml), and the acidic aqueous solution is directly subjected to high-performance liquid chromatography. Accuracy of measurement was high and the minimum limit of detection was 3 ng (S/N=3). There was a significant correlation between dosage and serum concentration (amoxapine plus 8-hydroxyamoxapine) at the steady state for patients receiving amoxapine (r=0.735,n=46,P<0.05). This method proved superior in analytical conditions, pretreatment to procedure for liquid chromatography, and accuracy to the conventional method. Therefore, this method is considered applicable for routine examination.
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