Autoanalysis of Urinary Stone-rlated Facors and Urinary Saturation Lebeles of Stone Compositions

尿路结石相关因素和尿路结石成分饱和度的自动分析

• 批准号: 09671644
• 负责人: HIRANO Atsuyuki
• 金额: $ 2.3万
• 依托单位: Wakayama Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671644/
• 关键词: renal calculus; capillary electrophoresis; urinary oxalate; urinary citrate; plasma oxalate; renal oxalate clearance; urinary cations; 蓚酸; クエン酸; 過蓚酸尿症; 家族発生

Urinary oxalate and citrate are the important promoter and inhibitor for urinary stone formation. Simultaneous dermination of both anions was established by capillary electrophoresis (CE) in this study. A Waters Quanta 4000E was used. Data was acquired on a microcomputer with Millennium soft ware from Waters. The electrolyte solution contained 25mM de-sodium hydrogen phosphate and 0.5mM anion BT. The coefficient variation and the recovery were excellent. The measurements of both anions by CE agreed with the values of oxalate by inochromatography and citrate by enzymatic procedure.Oxalate metabolism has been noted to play an important role on urinary stone formation. We tried, therefore, the determination of plasma oxalate using CE. The determination of plasma oxalate needs a high sensitivity because of its extremely low concentration, and to minimize oxalogenesis in vitro from ascorbic acid and glyoxylate. In our method, a larger caliber of capillary, a longer injection time of sample … More and a lower hold of sample dilution were adopted than in urinary oxalate determination. To minimize oxalogenesis, blood samples were ultrafiltrated under the lower temperature, then ultrafiltrates were rapidly acidified with a solution of 300mM boric acid and 50mM phosphoric acid (pH 2.0). The minimum detectable limit of plasma oxalate was 0.9μM, the coefficient variation was 5.8-16.0% and the recovery rate was 102% (88-126%). The plasma oxalate levels in 8 adults males were 2.39±1.46μM (Mean±SD), which agreed with the values in the recent repots.As a practical application of plasma oxalate determination, we measured renal oxalate clearance (Cox). Cox 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 (Cox/Ccr) ratios in each group were 0.70±0.16 and 1.11±0.34 respectively. Cox with 24 hours method were significantly higher than with one hour method, possibly because of an influence of dietary oxalate. Therefore, one hour method is advisable for Cox measurements. Which the Cox/Ccr ratios were higher than 1.0, suggests that oxalate would be excreted from the renal tubules.Although the determination of urinary anions by CE had been already established, we tried to measure NH4ィイD1+ィエD1, KィイD1+ィエD1, CaィイD1++ィエD1, NaィイD1+ィエD1, MgィイD1++ィエD1 in urine by this method. Differed from the oxalate and citrate determination, the system was used with a negative supply. The electrolyte solution contained 2mM crown ether/18-crown-6, 65mM HIBA and 5mM UV Cat-1. Separations of each cation peak in 1000 holds diluted urine samples with distilled water were satisfactory. Recoveries of each added 50μM cation were 90.0%, 88.1%, 75.6%, 102.0% and 100.8%. Less

尿草酸盐和柠檬酸盐是泌尿系结石形成的重要促进剂和抑制剂。建立了毛细管电泳法同时测定两种阴离子的方法。使用沃茨Quanta 4000 E。在具有来自沃茨的Millennium软件的微型计算机上获得数据。电解质溶液含有25 mM磷酸氢二钠和0.5mM阴离子BT。变异系数和回收率良好。这两种阴离子的测定值与草酸根离子色谱法和柠檬酸根酶法测定值一致，草酸根代谢在尿结石形成中起重要作用。因此，我们尝试用CE测定血浆草酸盐。血浆草酸的测定需要高灵敏度，因为它的浓度极低，并尽量减少体外抗坏血酸和乙醛酸的草酸生成。在我们的方法中，毛细管口径越大，进样时间越长， ...更多信息 采用较低的样品稀释度。为了最大限度地减少草酸生成，在较低温度下对血液样品进行超滤，然后用300 mM硼酸和50 mM磷酸（pH 2.0）的溶液快速酸化超滤。血浆草酸的最低检测限为0.9μM，变异系数为5.8-16.0%，回收率为102%（88-126%）。8例成年男性血浆草酸浓度为2.39±1.46μM（Mean±SD），与文献报道的结果一致，并测定了肾草酸清除率（考克斯）。1小时法考克斯曲线6例正常对照组为72.9±20.0 ml/min，8例结石组为83.2±27.8 ml/min。各组草酸/肌酐清除率（考克斯/Ccr）比值分别为0.70±0.16和1.11±0.34。24小时法的考克斯显著高于1小时法，这可能与日粮中草酸的影响有关。因此，考克斯测定宜采用一小时法。虽然尿阴离子的CE测定法已建立，但我们尝试用该方法测定尿中考克斯_4 ~（2-）D_1 + N_（2-）D_1、K ~（2-）D_1 + N_（2-）D_1、Ca ~（2-）D_1 + N_（2-）D_1、Na ~（2-）D_1 + N_（2-）D_1、Mg ~（2-）D_1 + N_（2-）D_1。与草酸盐和柠檬酸盐测定不同，该系统使用负电源。电解质溶液含有2 mM冠醚/18-冠-6，65 mM HIBA和5 mM UV Cat-1。用蒸馏水稀释尿样1000次后，各阳离子峰的分离令人满意。各50μM阳离子的回收率分别为90.0%、88.1%、75.6%、102.0%和100.8%。少