螢光法による血漿11-Hydoroxy corticosteroid測定法に関する研究

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In the past several years a number of methods have been developed for measuring corticosteroid levels in human plasma. De Moor et al. have used fluorescent technique to measure the free plasma 11-OHCS in human plasma. The procedure described here is based on their method, but minor modifications were necessary before satisfactory results were obtained. Using a domestic inexpensive fluorometer, the exciting light was provided by a mercury lamp with a 436mμ filter and a narrow band secondary filter of 530mμ was also used. All the reagents, available in our, country, were used without any further preliminary purification. 4ml plasma samples were needed for a valid determination, and it seems likely that specificity is improved when fluorescence is measured ten minutes after the addition of the reagent. It was demonstrated that the extraction with methylene chloride must be performed by gentle inversion, avoiding vigorous shaking. The reliability of the method has been assessed by criteria suggested by Brown et al. The estimates of precision and sensitivity are based on duplicate determination. The degree of precision compared with that obtained by de Moor et al., and the smallest measurable value which can be distinguished from zero was 1.7 μg/dl, if duplicate samples were used. When known amounts of cortisol were added to normal plasma, the mean recovery was 93%. The non-specific fluorescence of plasma samples was calculated according to the method of Moncloa et al., the mean value corresponded to 1.8 μg cortisol/dl plasma. The present method has been used successfully in studies on diurnal variation of plasma corticosteroid levels, in ACTH stimulation tests and in cortisol disappearance tests. It is therefore concluded that this method is a practical and reliable method for the determination of plasma 11-OHCS levels for routine clinical application.
千葉大学の論文
1967-01-28

螢光法による血漿11-Hydoroxy corticosteroid測定法に関する研究

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