de Faria L C, Pasquini C, de Oliveira Neto G
Instituto de Química, Universidade Estadual de Campinas CEP 13081, São Paulo, Brazil.
Analyst. 1991 Apr;116(4):357-60. doi: 10.1039/an9911600357.
A flow injection method was developed, aimed at the determination of urea in human serum. The system makes use of the naturally immobilized urease present in Canavalia ensiformis DC (jack bean). A column is filled with small pieces of this bean, and the sample (50 microliters) containing urea passes through it carried by a 1% NaCl solution. On leaving the column the stream is merged with an alkaline reagent (0.5 mol dm-3 NaOH; 0.5% disodium dihydrogen ethylenediaminetetraacetate). The ammonium ions, arising from the enzymatic reaction that occurs inside the column, are changed into the molecular form, which permeates a polytetrafluoroethylene membrane and is received in a de-ionized water acceptor stream. The ammonia ionizes causing an increase in the conductance, which is proportional to the urea content of the sample. About 40 samples can be processed in 1 h with negligible carry-over and with a relative standard deviation of 1% or less. The results are in agreement with those obtained by a standard spectrophotometric method.
开发了一种流动注射法,用于测定人血清中的尿素。该系统利用了刀豆(Canavalia ensiformis DC)中天然固定化的脲酶。将一根柱子填充上这种豆子的小块,含有尿素的样品(50微升)在1%氯化钠溶液的带动下通过柱子。离开柱子后,液流与一种碱性试剂(0.5摩尔/立方分米氢氧化钠;0.5%乙二胺四乙酸二钠)合并。在柱子内部发生的酶促反应产生的铵离子转变为分子形式,该分子形式透过聚四氟乙烯膜并被去离子水接收流接收。氨发生电离导致电导率增加,电导率与样品中的尿素含量成正比。1小时内可处理约40个样品,残留可忽略不计,相对标准偏差为1%或更低。结果与通过标准分光光度法获得的结果一致。