Archives of Pharmacal Research

The Pharmaceutical Society of Korea (대한약학회)
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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

  • Lee, Jung-Sook (Screening and Toxicology Center, Korea Research Institute of Chemical Technology) ;
  • Lee, Heeyong (Screening and Toxicology Center, Korea Research Institute of Chemical Technology) ;
  • Lee, Hye-Suk (Screening and Toxicology Center, Korea Research Institute of Chemical Technology) ;
  • Lee, Kang-Choon (College of Pharmacy, Sungkyunkwan University)
  • Published : 1994.10.01
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Abstract

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 $(25-40{\;}{\mu}m)$ 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 $C_{18}$ 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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