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KMID : 0043319940170050360
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Archives of Pharmacal Research
1994 Volume.17 No. 5 p.360 ~ p.363
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On-line Trace Enrichment for the Determination of Insulin in Biological Samples Using Reversed-Phase High Performance Liquid Chromatography with Column Switching
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Lee Jung-Sook
Lee Hee-Yong
Lee Hye-Suk
Lee Kang-Choon
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Abstract
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Column--swtiching technique with a reversed-phase high performance liquid chromatographic method has been developed for the routine analysis of radioiodinated insulin and its degadation products in biological fluids. The diluted biological samples were loaded onto a precolumn packed with LiChrosorb RP-8 using 0.1% trifuoroacetic acid (TFA) in water as a washing solvent. After valve switching, the concentrated insulins were eluted in the back-flush mode and separated by a W-Porex column with a gradient of 0.1% TFA in water and 0.1% TFA in acetonitrile as the mobile phase. The method showed good precision, accuracy and speed with the detection limit of 20 pg/ml. Total analysis time per sample was about 40 min and the coefficients of variation were less than 8, 2%.
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KEYWORD
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Column-switching technique, RP-HPLC, Insulin, Biological samples
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