Measurement of Total Plasma Homocysteine in Patients with Chronic Renal Failure Using HPLC/FLD

만성신부전증환자에서 HPLC/FLD를 이용한 혈장 Homocysteine의 측정

  • Kyung-Ok Lee (Department of Biochemistry, Seoul Medical Science Institute, Seoul Clinical Laboratories (SCL)) ;
  • Bo-Kyung Kang (Department of Biochemistry, Seoul Medical Science Institute, Seoul Clinical Laboratories (SCL)) ;
  • Hyung-Jin Roh (Department of Biochemistry, Seoul Medical Science Institute, Seoul Clinical Laboratories (SCL)) ;
  • Kwang-Suk Ryoo (Department of Biochemistry, Seoul Medical Science Institute, Seoul Clinical Laboratories (SCL)) ;
  • Jeong-Yeon Yoo (Department of Biochemistry, Seoul Medical Science Institute, Seoul Clinical Laboratories (SCL)) ;
  • Man-Jeong Paik (Department of Biochemistry, Seoul Medical Science Institute, Seoul Clinical Laboratories (SCL)) ;
  • Kang-Hyeob Lee (Department of Biochemistry, Seoul Medical Science Institute, Seoul Clinical Laboratories (SCL))
  • Published : 1997.06.01

Abstract

Cardiovascular disease has been the leading cause of death among patients with chronic renal failure. Many reports have been described that homocysteine is one of the independent risk factor to the occulsive vascular disease. In this study, HPLC/FLD (high performance liquid chromatography-fluorescence detector) technique was used to measure homocysteine level in patients with chronic renal failure and normal control group. The detection limit and recovery of total plasma homocysteine using HPLC/FLD were 98.6$\pm$5.8% and 0.2 nmol/ml, respectively. The linearity of this method was established in concentration range of 2~50 nmol/ml (correlation coefficient=0.9997). The concentrations of total plasma homocysteine were 6.81$\pm$1.54 nmol/ml and 27.28$\pm$14.94 nmol/ml in normal control (n=20) and patient group (n=90), respectively (p<0.05). In this study, the HPLC/FLD method showed high sensitivity and reproducibility for a routine clinical laboratory testing. Moreover determination of homocysteine level in plasma might be useful for a biochemical marker for predicting the cardiovascular diseases and for monitoring of therapeutic effect of lowering homocysteine in patients with chronic renal failure.

본 연구에서는 한국인 만성신부전증 환자에서 심혈관계 질환의 조기진단을 위한 생화학적 표지 자로서 homocysteine의 임상적 유용성을 검토하기 위하여, 만성신부전증 환자와 건강인에서 HPLC/FLD (high performance liquid chromatography/fluorescence detector)를 이 용하여 혈장 homocysteine 농도를 측정하고 그 결과를 비교하였다. 본 실험방법의 회수율은 98.6$\pm$5.8%를 나타내었으며, 0.2 nmol/L 보다 낮은 농도까지 측정이 가능하였고, 2~50 nmol/ml까지 직선성이 성립하였다 (correlation coefficient =0.9997). 한국인 중 건강한 정상인 (20명)과 만성신부전증 환자 (90명)에서 혈장 homocysteine은 각각 6.81$\pm$1.54 nmol/ml과 27.28$\pm$14.94 nmol/ml이었으며 , 환자군에서는 정상인군에 비하여 약 4배정도 높은 수치를 나타내었다 (p<0.05).본 실험의 결과로 볼 때 HPLC/FLD를 이용한 homocysteine 측정은 예민도와 재현성이 높아 routine 실험실 방법으로 유용성이 높을 것으로 생각되며, 또한 혈장 homocysteine의 측정은 만성신부전증 환자의 주요한 사망원인이 되는 폐쇄성 동맥질 환을 조기에 진단하거나, 혹은 homocysteine 농도를 저하시키는 치료를 실시 한 후 치료효과를 판단하기 위한 생화학적 marker로 활용될 수 있을 것으로 기대된다.

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