Measurement of δ-Aminolevulinic Acid in Urine by Fluorometric HPLC and Colorimetric Methods

비색법과 HPLC 법에 의한 요중 δ-Aminolevulinic acid의 측정치 비교

  • Ahn, Kyu-Dong (Department of Preventive Medicine, Medical College, Soonchunhyang University) ;
  • Yeon, You-Yong (Institution of Industrial Medicine, Soonchunhyang University) ;
  • Lee, Byung-Kook (Institution of Industrial Medicine, Soonchunhyang University)
  • 안규동 (순천향대학교 의과대학 예방의학교실) ;
  • 연유용 (순천향대학교 산업의학연구소) ;
  • 이병국 (순천향대학교 산업의학연구소)
  • Published : 1994.05.31

Abstract

The urinary excretion of ${\delta}$-aminolevulinic acid has been widely used as a measure of the biological effect of lead in lead exposed workers. It is usually measured by colorimetric method based on the color reaction of ALA-pyrrole with Ehrlich's reagent. But the results of ${\delta}$-ALA in urine by this method are somewhat artificially higher than expected due to the urinary ALA-like compound such as aminoacetone. On the other hand, the recently developed fluorometric HPLC method is very sensitive and specific for the measuring urinary ALA. In order to compare the data obtained by two methods and to investigate the interrelation between two methods, 117 lead workers with different lead exposure were checked urinary ${\delta}$-ALA, blood lead and other lead exposure related indices. The results obtained are as follows; 1. Urinary excretion of ${\delta}$-ALA by colorimetric method is 2.15 times higher than HPLC method in overall, revealing 2.47 times in workers of blood lead less than $20{\mu}g/dl$, 2.53 times in workers of blood lead $21-40{\mu}g/dl$ and 1.86 times in workers of blood lead over $41{\mu}g/dl$, respectively. 2. While the correlation coefficients of ${\delta}$-ALA measured by colorimetric method with blood lead and blood ZPP was 0.571 and 0.629, those of ${\delta}$-ALA measured by HPLC with blood lead and blood ZPP were 0.6l0 and 0.637. All the correlation coefficients were statistically significant, but there was no statistical difference of correlation coefficients between two methods. 3. The correlation coefficient of urinary excretion of ${\delta}$-ALA between two method was 0.838 without any correction, but it was 0.852 with the correction of specific gravity 1.024. 4. Simple linear regression of ${\delta}$-ALA measured by HPLC method on ${\delta}$-ALA measured by colorimetric method was (ALA-UPH)=-0.245+0.536 (ALA-UCO) without any correction and it was (SP ALA)=-0.525+0.598 (SP ALA-UCO) with the correction of specific gravity 1.024. With above results, it is recommended that the diagnostic criteria of ${\delta}$-ALA for lead poisoning needed to be revised if ${\delta}$-ALA is measured by HPLC rather than colorimetric method.

연폭로자에 있어서 연에 의한 생물학적 영향을 평가하는데는 요중 ${\delta}$-ALA 배설량이 아주 좋은 지표로 이용된다. 현재 이것은 Ehrlich's reagent와 ALA-pyrrole의 증색반응으로 측정하는 비색법이 주로 사용된다. 그러나 이 방법은 aminoacetone 같은 ALA 유사물질에 의하여 요중 배설량이 과다하게 측정된다. 한편 최근 개발된 fluorometric HPLC에 의한 요중 ${\delta}$-ALA 배설량의 분석은 아주 예민하고도 특이적인 것으로 알려져 있다. 따라서 본 연구는 혈중연과 다른 연폭로 지표를 측정하여 연폭로 수준이 다른 117명의 피검자에서 비색법과 HPLC 법에 의한 ${\delta}$-ALA 배설량을 측정하여 두 방법 간의 관련성을 조사하기 위하여 시도하였으며 결과는 다음과 같다. 1. 비색법에 의한 요중의 ${\delta}$-ALA 배설량은 HPLC 법에 비하여 전체적으로 2.15배, 혈중연 $20{\mu}g/dl$ 이하에서 2.47배, $21-40{\mu}g/dl$에서 2.53배, $41{\mu}g/dl$ 이상에서는 1.86배 높은 것으로 측정되었다. 2. 비색법에 의한 요중 ${\delta}$-ALA 배설량과 혈중연 및 혈중 ZPP 농도와의 상관은 0.571, 0.629였으며, HPLC 법은 0.610, 0.637로 통계적 유의성이 있었다(p<0.01). 3. 비색법과 HPLC 법에 의한 요중 ${\delta}$-ALA 배설량 간의 상관은 0.848 이었으며, 요비중으로 보정하였을 때는 0.852였으며 통계적으로 유의하였다(p<0.01). 4. HPLC 법에 의한 요중 ${\delta}$-ALA 배설량을 종속변수로 하고 비색버버을 독립변수로 하였을 때 희기방정식은 (ALA-UHP)=-0.245+0.536 (ALA-UCO)(p<0.0001)였으며, 요비중 1.024로 보정하였을 때의 희기방정식은 (SP ALA-UHP)=-0.525+0.599 (SP ALA-UCO)(p<0.0001)였다. 상기와 같은 결과에서 현행 산업안전보건법 시행규칙에서 연중독 진단을 행하는 경우 HPLC로 요중 ${\delta}$-ALA 배설량을 측정한다면 연중독 진단 기준의 개정이 필요할 것으로 생각된다.

Keywords