Peng Z, Wang C, Li C, Gao Q, Wang Z, Zhang G, Zhang J, Li X, Peng J
Central Hospital of Jingzhou City, Hubei Province, Jingzhou, 434100.
Se Pu. 1998 Mar;16(2):146-8.
In this paper a sensitive, rapid and simple HPLC method for the simultaneous determination of creatinine (Cr), pseudouridine (Pu) and uric acid(UA) has been established. We have evaluated clinical value of the method in the early diagnosis of diabetic nephropathy (DN). Separation was obtained using Shim-pack CLC-ODS 15 x 0.6 cm column and mobile phase of buffer solution of phosphate (pH 3.0, 0.02 mol/L) with flow rate of 1 mL/min. Detection was performed with UV detector at an automatic adjustment of wavelength. The recoveries of Cr, Pu and UA were 101.4%, 104.9% and 105.0% respectively. The calibration curves were linear within the concentration range of 8.6-274.6 mumol/L for Cr, 0.72-22.93 mumol/L for Pu and 19.1-612.7 mumol/L for UA (n = 6, r > 0.999, p < 0.01). The CV for within day and between day measurements were < 2.5% and < 5.0% respectively. In addition, Pu, Cr and UA were simultaneously determined in serum and urine of 39 patients with diabetic mellitus (DM). 15 patients with nephrotic syndrome (NS) and 53 normal subjects by the method. Results include: 1. the levels of serum Pu in all DM patients (100%) with microalbuminuria (MiAU) and macroalbuminuria (MaAU) were greater than maximum of normal subjects (X + 2S). In addition, in 9 DM patients (41%) with normal albuminuria (NAU) the levels were also greater than the maximum. 2. In patients with DM, there was no correlation between the serum Pu and the urinary albumin excretion (UAE), whereas the serum Pu correlated closely with the serum Cr. However, the incidence for serum Pu increase was significantly greater in patients. 3. The levels of serum Pu in patients with NS were greater than those in control subjects and patients with DM. Conclusion is that the determination of serum Pu could be used as sensitivity index for the early diagnosis of DN.
本文建立了一种灵敏、快速且简便的高效液相色谱法，用于同时测定肌酐（Cr）、假尿苷（Pu）和尿酸（UA）。我们评估了该方法在糖尿病肾病（DN）早期诊断中的临床价值。采用Shim - pack CLC - ODS 15×0.6 cm色谱柱，以磷酸盐缓冲溶液（pH 3.0，0.02 mol/L）为流动相，流速为1 mL/min进行分离。使用紫外检测器在自动波长调节下进行检测。Cr、Pu和UA的回收率分别为101.4%、104.9%和105.0%。Cr的校准曲线在8.6 - 274.6 μmol/L浓度范围内呈线性，Pu在0.72 - 22.93 μmol/L浓度范围内呈线性，UA在19.1 - 612.7 μmol/L浓度范围内呈线性（n = 6，r > 0.999，p < 0.01）。日内和日间测量的变异系数分别< 2.5%和< 5.0%。此外，采用该方法同时测定了39例糖尿病（DM）患者、15例肾病综合征（NS）患者和53例正常受试者的血清及尿液中的Pu、Cr和UA。结果如下：1. 所有微量白蛋白尿（MiAU）和大量白蛋白尿（MaAU）的DM患者（100%）血清Pu水平均高于正常受试者最大值（X + 2S）。此外，9例正常白蛋白尿（NAU）的DM患者（41%）血清Pu水平也高于最大值。2. 在DM患者中，血清Pu与尿白蛋白排泄率（UAE）无相关性，而血清Pu与血清Cr密切相关。然而，患者血清Pu升高的发生率显著更高。3. NS患者血清Pu水平高于对照组和DM患者。结论是血清Pu的测定可作为DN早期诊断的敏感指标。
