Trace determination and characterization of some proteins in blood serum by high-performance liquid chromatography with a time-resolved laser fluorimetric detector |
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Authors: | Yuji Kawabata Kouji Sauda Totaro Imasaka Nobuhiko Ishibashi |
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Affiliation: | Faculty of Engineering, Kyushu University, Hakozaki, Fukuoka 812 Japan |
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Abstract: | Protein in human serum is measured by high-performance liquid chromatography with a time- resolved laser fluorimetric detector. The detection limit is 2.3 pmol for bovine serum albumin; the value is governed by fluctuation of the background fluorescence from free 1-anilino-8-naphthalene sulfonate (ANS) used as the fluorescence probe. Micropolarity of the binding site in protein is estimated by measuring the fluorescence lifetime of ANS bound to protein; the micropolarity for albumin is lower than those for α- and γ-globulins. |
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