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标题：A Modified Fluorescence Polarization Immunoassay Method Incorporating Fat Emulsion (FE-FPIA) to Determine Cyclosporin A Concentrations in Rat Skin
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作者：Yuji KUROSAKI ; Tomoya ASADA ; Nanae YASUMOTO 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：1999
卷号：22
期号：5
页码：539-542
DOI：10.1248/bpb.22.539
出版社：The Pharmaceutical Society of Japan
摘要：We have developed a simple, sensitive and reliable assay procedure for cyclosporin A (CyA), a modified fluorescence polarization immunoasay method incorporating fat emulsion (FE-FPIA), to determine the CyA content in rat skin. The conventional fluorescence polarization immunoassay (FPIA) method for CyA using a commercially available FPIA kit, TDX^[○!R] cyclosporine monoclonal whole blood, was modified. A fat emulsion for intravenous infusion, Intralips^[○!R], was incorporated for dissolving the CyA extracted from the skin tissue, and a mixture of MeOH/purified water was used as the sample pretreatment medium instead of the precipitation reagent in the conventional FPIA kit intended for whole blood samples. These modifications enabled us to produce a reliable and the sensitive assay of CyA in skin tissue. The reproducibility (coefficient of variation), detection limit, and assay time for FE-FPIA were below 2%, 25 ng/ml, and about 24 min/24 samples, respectively, and were comparable with those for the whole blood samples determined by the conventional FPIA. Pre-purification of samples required by the HPLC assay is not needed in the FE-FPIA method. The usefulness of the FE-FPIA method in evaluating the topical pharmacokinetics of CyA in skin is discussed.
关键词：cyclosporin A;fluorescence polarization immunoassay;fat emulsion;topical pharmacokinetics