高效毛细管电泳法同时测定尿液清蛋白和肌酐

目的建立同时测定尿液清蛋白和肌酐的高效毛细管电泳法。方法对缓冲系统、pH值、缓冲液的离子强度、检测波长、表面活性剂、电压及电流和进样方法等进行研究,建立同时测定清蛋白和肌酐的毛细管电泳法。结果选择20mmol/L、pH9.3硼砂-NaOH(含6mmol/LSDS)缓冲液作为毛细管区带电泳运行液,利用外标法定量,检测波长214nm,同时测定肌酐和清蛋白。非涂层毛细管有效长度47.5cm,内径75μm;电流79μA,压力进样1psi、4s,电泳时间为12min。肌酐和清蛋白的线性范围分别为4.32～8840μmol/L和1.95～1000mg/L,肌酐和清蛋白最低检出限分别为0.977μmol/L和0.285mg/L。本法的日内和日间变异系数均小于4.8%。肌酐和清蛋白的平均回收率分别为99.6%和102.4%。结论建立的方法线性范围宽,测定简单快速,可用于临床检测。     

Measurement of the albumin content of urinary protein using dipsticks.

An albumin selective urine strip based on bis (3',3'-diiodo-4', 4'-dihydroxy-5',5'-dinitrophenyl)-3,4,5,6-tetrabromo sulfonphthalein dye (DIDNTB) dye was examined in populations with clinical proteinuria. The relationship of albumin to the sum concentration of all protein in urine was found to vary widely even though the albumin concentration generally increased with the total protein concentration. The albumin reagent strips correlated well with immuno-nephrometric assays for albumin on specimens from hypertensives, diabetics, and renal disease which tended to have albumin contents of >/= 50.0%. High proteinuria concentrations of > 250 mg/l, with low albumin contents of 相似文献   

Clinical analysis by microchip capillary electrophoresis

Clinical analysis often requires rapid, automated, and high-throughput analytical systems. Microchip capillary electrophoresis (CE) has the potential to achieve very rapid analysis (typically seconds), easy integration of multiple analytical steps, and parallel operation. Although it is currently still in an early stage of development, there are already many reports in the literature describing the applications of microchip CE in clinical analysis. At the same time, more fully automated and higher throughput commercial instruments for microchip CE are becoming available and are expected to further enhance the development of applications of microchip CE in routine clinical testing. To put into perspective its potential, we briefly compare microchip CE with conventional CE and review developments in this technique that may be useful in diagnosis of major diseases.     

Rapid heteroduplex analysis by capillary electrophoresis

The very important parameters for mutation screening are short time of analysis and high throughput. The analytical platform which fulfills these criteria most satisfactorily is capillary electrophoresis. Here we show the influence of several parameters such as temperature, presence of glycerol, capillary length and polymer concentration on the electrophoretic properties of DNA duplexes and evaluate their contribution to the overall time of analysis. The careful optimization of analyzed conditions allowed us to significantly decrease the time required for the detection of the 185delAG and 4153delA mutations by heteroduplex analysis. It enabled us to analyze these typical BRCA1 gene deleterious mutations in several minutes only by using very popular and widely accessible capillary electrophoresis instrumentation.     

Quantification of urinary allantoin by capillary zone electrophoresis during recombinant urate oxydase (rasburicase) therapy

OBJECTIVES: Rasburicase (Fasturtec) is used to prevent or treat hyperuricemia associated with chemotherapy. We developed a capillary zone electrophoresis method to measure urinary allantoin, the degradation product of uric acid by rasburicase. DESIGN AND METHODS: Electrophoresis was performed using a P/ACE 5500 system (Beckman) with a fused silica capillary tube and a UV-visible detector set at 214 nm. Urine samples from 10 patients with non-Hodgkin's lymphoma were analyzed to validate the technique. RESULTS: Using a sodium tetraborate running buffer, urinary allantoin was separated from related compounds and internal standard in less than 30 min. The method was linear up to 1.25 g/L (quantification limit: 30 mg/L); precision was below 10%. The total amount of allantoin excreted in patients treated by rasburicase ranged from 1.5 g to 7.9 g/4 days. CONCLUSION: This CZE assay is a simple, rapid and reproducible method to measure allantoin in urine. Different elimination profiles have been found in patients treated with rasburicase.     

毛细管电泳法和微芯片电泳法在糖类分析中的应用

糖类是三大生命物质之一,它参与多种生物代谢和其他生命活动.由于糖类的结构复杂,对它的研究方法要求较高.毛细管电泳以其快速、高效和高灵敏度、所需样品少等特点被广泛应用于糖类化合物的分析.而近几年快速发展的微芯片技术也在糖类分析中得到应用.本文就毛细管电泳法和微芯片电泳法在各类糖中的应用进行了阐述.     

毛细管电泳法分析糖基化血红蛋白

目的：探讨毛细管电泳在分析糖基化血红蛋白水平方面的临床应用价值。方法该文利用长30 cm、内径25μm 的未涂层熔融石英毛细管,利用血红蛋白在415 nm 波长处的特异性吸收峰,二极管阵列（PDA）法检测健康人与糖尿病及高危人群中的糖基化血红蛋白（HbA1c）水平。在研究过程中,对精密度和准确度进行了精确分析。结果 HbAlc 和非 HbAlc 成功分离,峰型清晰锐利且能对 HbAlc 进行定量分析。结论毛细管电泳法具有特异性好、精密度和准确度高等优点,因此,该法应该在临床上被推广应用。     

血清蛋白的毛细管电泳分析

采用毛细管电泳技术建立了血清蛋白毛细管自由区带电泳分析方法，并与醋酸纤维薄膜电泳法相比较。利用ｐＨ９．８、１００ｍｍｏｌ／Ｌ硼酸缓冲液作电泳缓冲液，未涂渍毛细管２１ｃｍ×５０μｍ（ｉ．ｄ），检测波长２００ｎｍ，当电压为０．５ｋＶ／ｃｍ时，在５分钟内可将血清蛋白分为五条蛋白区带。利用ｐＨ８．５、２５ｍｍｏｌ／ＬＳＤＳ、２０ｍｍｏｌ／Ｌ硼酸缓冲液作样品稀释液，未涂渍毛细管４５ｃｍ×５０μｍ（ｉ．ｄ），在电压２２．５ｋＶ条件下，可将血清蛋白分离１０条区带。该方法稳定、重复性好。结果表明毛细管自由区带电泳可作为临床实验室分析血清蛋白常规分析方法     

毛细管区带电泳法拆分氨甲蝶呤手性对映体

目的建立一种快速拆分氨甲蝶呤手性对映体的方法。方法用毛细管区带电泳技术，以中性环糊精作为氨甲蝶呤拆分选择剂，对环糊精浓度、缓冲液浓度、pH值、电泳温度和改性剂浓度等分离条件进行优化。结果用含25mmol／L的羟丙基-8-环糊精、10％的异丙醇、80mmol／L(pH7．4)的磷酸盐缓冲液，在18kV分离电压，毛细管温度25℃时，可使氨甲蝶呤对映体在25min内达到基线分离，分离度为1．51。结论高效毛细管区带电泳法拆分氨甲蝶呤手性对映体简单、快速、经济。     

Iohexol in serum determined by capillary electrophoresis.

Iohexol, a nonionic compound used as a contrast medium for angiography and as a measure of the glomerular filtration rate, was quantified in serum by capillary electrophoresis. Comparable results were obtained for serum samples deproteinized with acetonitrile or analyzed directly after 50-fold dilution with borate buffer. Serum samples were electrophoresed for 2.6 min at 12 kV in a borate buffer with detection at 254 nm and with 3-isobutyl-1-methylxanthine as internal standard. Acetonitrile deproteinization gave a greater sensitivity than did sample dilution. Between-run CVs were between 4.7% and 6.7%, and within-run CVs were between 2.5 and 3.2%. Analytical recoveries were 95-105%. Results of the method compared well with those by high-performance liquid chromatography (slope 0.96, intercept 0.005 g/L). This method demonstrates the potential of capillary electrophoresis for rapid and simple quantification of small molecules.     

Routine differential diagnosis of proteinurias by capillary electrophoresis.

Capillary electrophoresis is a relatively new analytical technique that begins to have an impact on both routine and research in clinical laboratories. Recently, a fully automated system has become commercially available (Paragon CZE 2000, Beckman, USA) for the analysis of human serum proteins. Urine protein analysis, on the other hand, is currently accomplished by electrophoresis of concentrated urine specimens. The method is used to distinguish the glomerular from the tubular proteinuria and for the identification of Bence-Jones proteins. The procedure is labor-intensive and technically demanding. We developed a technique for the serum capillary electrophoresis instrument that can also be applied routinely to the differential diagnosis of proteinurias. Overriding the programmed dilution step of the instrument, we were able to distinguish different types of proteinurias without concentration of specimens with a total protein content of 150-200 mg/l as determined by sulfosalicylic acid. The different electrophoretic patterns obtained by the capillary electrophoresis system for various specimens correlated well with established techniques (Hydragel Proteinurie Kit, Sebia, France). The method is applicable for routine analysis of urinary proteins. It is reliable, less expensive and faster than the conventional methods (electrophoretic or immunonephelometric) used today for the differentiation of proteinurias, and it can be used as a quick screening test.     

Determination of serum valproic acid drugs by capillary electrophoresis

Monitoring of valproates is caused by the nonlinear pharmacokinetics of the drugs and by the clinical effect-concentration dependence. Rapid and simple procedure has been developed to determine valproic acid in the serum and spinal fluid by capillary electrophoresis. Under the chosen conditions, the calibration curve is linear in the range of concentrations of 10 to 150 mg/l. The detection limit of valproate is 1.0 mg/ml. The accuracy of valproate determination was confirmed by the addition method. The findings were compared by the results of ion chromatographic determination that had been chosen as the reference technique. More than 80 serum samples from patients with epilepsy were analyzed.     

毛细管电泳法分离血清中乳酸脱氢酶同工酶

目的利用还原型辅酶I（NADH）在340nm波长处特异性吸收峰，建立用毛细管区带电泳在线检测人血清中乳酸脱氢酶（LDH）同工酶的方法。方法实验中采用未涂层毛细管，长60cm，内径75μm，pH9．4（23℃）二氨基二甲基1，3丙二醇（AM2P）缓冲液含20mmol/L氧化型辅酶I（NAD^＋）、51，6mmol/L乳酸锂。结果在25min内完成电泳分离，乳酸脱氢酶同工酶1（LDH1）、乳酸脱氢酶同工酶5（LDH5）纯品各自均可得到较好峰形，而且LDH1、LDH2纯品的混合物亦可得到完全分离，在分析正常人血清与肝癌患者血清时，其结果与美国Helena公司REP全自动琼脂糖电泳结果一致，取得满意效果。结论该法快速、低耗，具有较好的推广应用价值。