Dual-column high-performance liquid chromatographic urinary organic acid profiling |
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Authors: | DN Buchanan JG Thoene |
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Institution: | Genetic Screening Laboratory, Michigan Department of Mental Health, Hawthorn Center, 18471 Haggerty Road, Northville, Michigan 48167 USA |
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Abstract: | Two-dimensional high-performance liquid chromatography (HPLC) is increasingly used for the separation and identification of compounds in biological matrices. Conventional two-dimensional HPLC involves either heart-cutting or column-switching techniques. These techniques work well but are very time consuming when the analysis involves many compounds and requires more than a few minutes to completely elute from the column. We have modified the column-switching technique by utilizing two columns in series, an Aminex HPX-87H organic acid column followed by a 15-cm 5-μm C18 analytical column. Both columns are compatible with the isocratic pH 2.5 H2SO4 mobile phase employed in the organic acid profiling. The dual-column system affords better resolution of urinary organic acids than does either column separately. Reversing the column order does not dramatically affect the elution pattern (peak shape, peak height, and Rf values are approximately the same). The dualcolumn HPLC configuration works well as a rapid means of screening urinary carboxylic acids prior to subsequent definitive analyses of abnormal samples. |
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