Optimized fluorometric determination of urinary delta-aminolevulinic acid by using pre-column derivatization, and identification of the derivative.
Optimized fluorometric determination of urinary delta-aminolevulinic acid by using pre-column derivatization, and identification of the derivative.
复制标题
使用柱前衍生优化尿液 δ-氨基乙酰丙酸的荧光测定，并鉴定衍生物。
DOI：
10.1093/clinchem/36.8.1494
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发表时间：
1990
影响因子：
9.3
通讯作者：
R. Miura
中科院分区：
文献类型：
--
作者：
A. Okayama;S. Fujii;R. Miura
我们以前报道了有用的荧光法测定尿中δ-氨基乙酰丙酸（ALA）的浓度，通过使用柱后衍生监测铅暴露的影响。我们进一步改进了该方法，通过引入柱前衍生化，利用ALA与乙酰丙酮和甲醛的反应。血液中铅浓度低至162微克/升时，造血系统对铅暴露的反应现在可以很容易地检测出来。采用高效液相色谱法在十八烷基硅胶柱上分离了具有荧光的ALA衍生物2-亚甲基氨基-3，5-二乙酰基-4，6-二甲基苯基丙酸，用荧光光度计测定了其荧光强度。对12份铅作业工人尿样和对照组尿样的回收率为91.9-110.2%。柱前衍生法与柱后衍生法测定结果一致。新方法将检测限的灵敏度提高到每毫升尿液中10微克的δ-氨基乙酰丙酸，并且足够简单，可用于常规监测暴露于低浓度铅的生物效应。
We previously reported the usefulness of a fluorometric method to determine urinary delta-aminolevulinic acid (ALA) concentrations by using post-column derivatization to monitor the effect of lead exposure. We have further improved the method by introducing pre-column derivatization by using reaction of ALA with acetylacetone and formaldehyde. Response of the hematopoietic system to lead exposure can now be easily detected at blood lead concentrations as low as 162 micrograms/L. The fluorescent ALA derivative, a new aromatic product, 2-methylideneamino-3,5-diacetyl-4,6-dimethylphenylpropionic acid, is separated on octadecyl silica column by high-performance liquid chromatography and the fluorescence intensity is detected with a fluorophotometer. Sample recoveries for 12 urine samples from workers exposed to lead and unexposed controls were 91.9-110.2%. The results obtained by the pre-column derivatization method agreed with those by the post-column derivatization method. The new method increases the sensitivity to a detection limit to 10 micrograms of delta-aminolevulinic acid per milliliter of urine and is simple enough to be used for routine monitoring of the biological effect of exposure to low concentrations of lead.