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首页> 外文期刊>Life sciences >Radioimmunoassay of N-acetyl-N-formyl-5-methoxykynuramine (AFMK): a melatonin oxidative metabolite.
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Radioimmunoassay of N-acetyl-N-formyl-5-methoxykynuramine (AFMK): a melatonin oxidative metabolite.

机译：N-乙酰基-N-甲酰基-5-甲氧基犬尿嘧啶（AFMK）的放射免疫分析：褪黑素氧化代谢产物。

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摘要

N-acetyl-N-formyl-5-methoxykynuramine (AFMK) is a melatonin metabolite identified in rat brain by Hirata et al. (The Journal of Biological Chemistry 249 (1974) 1311). Since no assay has been described for its routine measurement, we have developed and validated such a radioimmunoassay.We synthesized AFMK and N-acetyl-5-methoxykynuramine (AMK), in order to produce anti-AFMK antibodies and to standardize the assay. The tracer [3H]-AFMK was obtained from [3H]-melatonin. The assay was preceded by a chromatographic step on Celite microcolumn in order to increase its specificity.The assay was suitable for the measurement of AFMK levels ranging from 59 to 1894 pmol/L. The detection limit of the assay was routinely set at 65 pmol/L. The intra- and inter-assay coefficients of variation were 3.5% and 11% respectively. Investigation of the 24 h plasma pattern in healthy volunteers did not reveal any AFMK levels in plasma samples. In rats, plasma AFMK showed a peak after melatonin injection, which confirmed the invivo AFMK production as a melatonin metabolite.This AFMK assay is suitable for studies on melatonin metabolism.
机译：N-乙酰基-N-甲酰基-5-甲氧基kyururamine（AFMK）是由Hirata等在大鼠脑中鉴定的褪黑激素代谢产物。 （Journal of Biological Chemistry 249（1974）1311）。由于没有描述用于常规测量的检测方法，因此我们开发并验证了这种放射免疫检测方法。我们合成了AFMK和N-乙酰基-5-甲氧基尿嘧啶（AMK），以产生抗AFMK抗体并使检测标准化。示踪物[3H] -AFMK获自[3H]-褪黑激素。为了提高特异性，该方法在Celite微柱上进行了色谱步骤，适用于测量59- 1894 pmol / L的AFMK水平。该方法的检测极限通常设置为65 pmol / L。批内和批间变异系数分别为3.5％和11％。对健康志愿者的24小时血浆模式进行的调查未发现血浆样品中的任何AFMK水平。在大鼠中，褪黑素注射后血浆AFMK出现一个峰值，这证实了体内AFMK作为褪黑素代谢产物的产生。该AFMK分析法适合进行褪黑素代谢研究。

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