Ogura H
Department of Urology, Wakayama Medical College.
Nihon Hinyokika Gakkai Zasshi. 2000 Jun;91(6):547-55. doi: 10.5980/jpnjurol1989.91.547.
The determinations of oxalate in urine and plasma are important in the evaluation and treatment of patients with calcium oxalate nephrolithiasis. Although many analytical methods for determining oxalate have been developed, most of them need complicated sample preparation, and are expensive for routine examination. Especially for estimation of plasma oxalate, much more sensitive measurement is required because of the extremely low concentration. A simple and rapid assay for oxalate in urine and plasma by capillary electrophoresis has been described here, and utilized for assessment of renal oxalate clearance. In addition, simultaneous determination of urinary oxalate and citrate was developed.
A Waters Quanta 4000E system was used with a detection at 185 nm. Separation was obtained on a fused silica capillary, 60 cm long x 75 microns and 100 microns (i.d.) for urine and plasma samples respectively. Urine samples were diluted with 60 mM hydrochloric acid, and ultrafiltrates of plasma were acidified and diluted with 300 mM boric acid and 50 mM phosphoric acid.
The intraassay coefficient variation was 2.7-4.0% for urinary oxalate, and 1.3-3.9% for citrate. The mean recovery ratio of 0.2 mM oxalate and 1.0 mM citrate added to 10 samples were 99.0% (92.6-107.4%) and 98.4% (91.2-103.9%), respectively. In the determination of plasma oxalate, the minimum detectable limit was 0.9 microM, the coefficient variation was 5.8-16.0%, and the recovery rate was 101.5% (87.8-125.6%). The plasma oxalate levels in 8 adult males were 2.39 +/- 1.46 microM (Mean +/- SD). Renal oxalate clearances with one hour method were 72.9 +/- 20.0 ml/min in 6 healthy controls and 83.2 +/- 27.8 ml/min in 8 stone formers. Oxalate/creatinine clearance ratios in each groups were 0.70 +/- 0.16 and 1.11 +/- 0.34 respectively.
The simultaneous determination of urinary oxalate and citrate was satisfactory. Capillary electrophoresis is suited for routine examination of urinary oxalate and citrate with the advantage on simplicity and economy. The assay of plasma oxalate by this method was also acceptably sensitive, specific under a low temperature and an acidification.
测定尿液和血浆中的草酸盐对于草酸钙肾结石患者的评估和治疗具有重要意义。尽管已经开发出许多测定草酸盐的分析方法,但大多数方法需要复杂的样品制备,且用于常规检查成本较高。特别是对于血浆草酸盐的测定,由于其浓度极低,需要更灵敏的测量方法。本文描述了一种通过毛细管电泳简单快速测定尿液和血浆中草酸盐的方法,并用于评估肾脏草酸盐清除率。此外,还开发了同时测定尿草酸盐和柠檬酸盐的方法。
使用Waters Quanta 4000E系统,检测波长为185nm。分别在60cm长、内径75μm和100μm的熔融石英毛细管上对尿液和血浆样品进行分离。尿液样品用60mM盐酸稀释,血浆超滤物用300mM硼酸和50mM磷酸酸化并稀释。
尿草酸盐的批内变异系数为2.7 - 4.0%,柠檬酸盐为1.3 - 3.9%。向10个样品中添加0.2mM草酸盐和1.0mM柠檬酸盐的平均回收率分别为99.0%(92.6 - 107.4%)和98.4%(91.2 - 103.9%)。在血浆草酸盐测定中,最低检测限为0.9μM,变异系数为5.8 - 16.0%,回收率为101.5%(87.8 - 125.6%)。8名成年男性的血浆草酸盐水平为2.39±1.46μM(平均值±标准差)。6名健康对照者用一小时法测得的肾脏草酸盐清除率为72.9±20.0ml/min,8名结石形成者为83.2±27.8ml/min。各组的草酸盐/肌酐清除率分别为0.70±0.16和1.11±0.34。
同时测定尿草酸盐和柠檬酸盐的结果令人满意。毛细管电泳适用于尿液草酸盐和柠檬酸盐的常规检查,具有简单经济的优点。用该方法测定血浆草酸盐在低温和酸化条件下也具有可接受的灵敏度和特异性。