The HPLC analysis of vanillil-almond, 5-hydroxiindolacetic, homovanillic and homogentisic acids in biologic fluids in clinical laboratory].

A A Dutov, Iu L Luk'ianova, E V Belen'kova, A V Sverkunova, O N Konovalova, E N Fedorova, A V Ermolina

Klinicheskaia laboratornaia diagnostika 2012 Feb

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The simple and fast HPLC technique of analysis of vanillil-almond, 5-hydroxiindolacetic, homovanillic and homogentisic acids in urine with solid-phase extraction using ultra cross-linked polystyrene (Purosep-200) is proposed. The separation on column Chromolith Performance RP-18e "Merck" 100x4.6 mm with monolithic phase-reversed silica gel in isocratic or gradient mode with ultraviolet detection under 285 nm. The isocratic mode is applied in case of ordinary analysis of vanillil-almond or homogentisic acids. The composition of eluent is isopropanol - water - TFA (1:99:0.025, v/v/v), flow speed is 1400 mkl/min; pressure is 37 bar, full separation less than in 4 min. The gradient low pressure mode is applied to analyze vanillil-almond, hydroxiindolacetic and homovanillic acids. Under this approach the switching to second eluent occurred from second minute. The composition of eluent is isopropanol-water-TFA (6:94:0.025, v/v/v), flow speed is 1400 mkl/min, pressure is 43 bar, full separation is less than in 7 min. The output (extraction share) consisted 78-113%. The simplicity reproducibility and sufficient sensitivity of technique in combination with the possibility of its application on standard chromatographic equipment (isocratic pump and ultraviolet detector) made it useful for routine clinical application.

Citation

A A Dutov, Iu L Luk'ianova, E V Belen'kova, A V Sverkunova, O N Konovalova, E N Fedorova, A V Ermolina. The HPLC analysis of vanillil-almond, 5-hydroxiindolacetic, homovanillic and homogentisic acids in biologic fluids in clinical laboratory]. Klinicheskaia laboratornaia diagnostika. 2012 Feb(2):28-31