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。     

L-proline assay in biological material by capillary zone electrophoresis

Conditions for determination of L-proline were optimized without prederivation by capillary zone electrophoresis. The pH value of buffer electrolyte, temperature, and the time of sample injection into the capillary were examined for their impact on the results of assay of L-proline. The latter was estimated in biological fluid samples. The duration of the assay was not more than 13 minutes.     

Separation of serum proteins by automated capillary zone electrophoresis.

Capillary zone electrophoresis (CZE) of serum proteins is increasingly gaining impact in clinical laboratories. Two automated multichannel instruments dedicated to the separation of serum proteins have become available over the last 6 years, the Paragon CZE 2000 (Beckman Coulter, CA, USA) and, more recently, the Capillarys (Sebia, France). This review focuses on the performance of these commercial instruments to separate serum proteins in a clinical laboratory setting. The utility of CZE to recognize various dysproteinemias and to detect and identify monoclonal proteins will be described and systematically reviewed. The reader will be provided with a summation and an understanding of CZE-specific interference.     

毛细管区带电泳测定血浆中丙戊酸钠浓度

目的建立毛细管区带电泳测定血浆中丙戊酸钠的方法。方法血浆标本经HCL酸化,正己烷萃取,再以NAOH溶液反提取丙戊酸钠。采用75ΜM(ID)×37CM石英毛细管柱分离。电泳缓冲液为15MMOL/L PH5.7水杨酸钠溶液(含0.5MMOL/L CTAB,15%乙醇),分离电压20KV,电泳时间6MIN,温度20℃,阴极压力进样5S,阳极检测,波长214NM。结果血药浓度线性范围25~200ΜG/ML,R=0.999,最低检测浓度0.35ΜG/ML(S/N=3),萃取回收率87.4%,方法回收率100.1%,日内、日间变异系数(CV)分别小于4%、6%。结论本方法可以简便、快速、灵敏地测定血浆中丙戊酸钠浓度。     

Determination of carbohydrate-deficient transferrin using capillary zone electrophoresis

BACKGROUND: Current methods for carbohydrate-deficient transferrin (CDT) often suffer from low precision, complexity, or risk of false positives attributable to genetic variants. In this study, a new capillary zone electrophoresis (CZE) method for CDT was developed. METHODS: CZE was performed on a P/ACE 5000 using fused-silica capillaries [50 microm (i.d.) x 47 cm] and the CEOFIX CDT buffer system with addition of 50 microL of anti-C3c and 10 microL of anti-hemoglobin. Native sera were loaded by high-pressure injection for 3 s, separated at 28 kV over 12 min, and monitored at 214 nm. RESULTS: CDT was completely resolved by differences in migration times (di-trisialotransferrin, 9.86 +/- 0.05 min; monosialotransferrin, 9.72 +/- 0.05 min; asialotransferrin, 9.52 +/- 0.04 min), with a CV of 0.15%. The number of theoretical plates was 312,000 +/- 21,000 for the mono- and 199 000 +/- 6500 for the di-trisialylated transferrin. Genetic CB and CD variants showed prominent peaks with migration times of 10.12 +/- 0.06 and 9.89 +/- 0.03 min, respectively, and the carbohydrate-deficient glycoprotein syndrome could be detected, excluding false-positive results. CZE results (as a percentage; y) correlated with the Axis %CDT TIA (x) values by Deming regression analysis: y = 1.92x - 7.29; r = 0.89. CDT values in 130 healthy nonalcoholics were determined. The 2.5th and 97.5th percentiles were 1.84% and 6.79%. CONCLUSIONS: CZE without sample pretreatment can determine CDT with good precision, allows detection of variants, and correlates with ion-exchange chromatography.     

应用高效毛细管电泳技术快速分离尿肌酐

目的　建立 1种快速测定尿中肌酐浓度的毛细管电泳方法。方法　用pH2 .5、0 .1mmol/L磷酸作为电泳缓冲液 ,利用非涂层石英毛细管 48.5cm× 5 0 μm(id) ,采用外标法定量 ,检测波长 191nm。 结果　本法在34 .5～ 8840 μmol/L浓度范围内呈良好的线性 (r =0 .998) ,日内和日间变异系数分别为 3.5 %、6 .2 %,平均回收率 94.7%。与苦味酸法有良好的相关性 (Y =1.0 0 4X 0 .0 0 4,r =0 .998,n =45 )。结论　本法线性范围宽、简单、快速 ,可应用于临床样品的检测。     

Determination of folic acid by capillary zone electrophoresis with indirect chemiluminescence detection

A capillary electrophoresis method with on-line inhibited chemiluminescence (CL) detection was first used to determine folic acid (FA). This method was established based on the quenching effect of FA on the CL reaction of luminol with a Ag(iii) complex in alkaline medium. The separation was conducted with a 20.0 mM sodium borate buffer containing 1.0 mmol L⁻¹ luminol. Under optimized conditions, FA was baseline separated and detected in less than 10 min. The limit of detection of FA was 1.3 mg L⁻¹, with a linear range of 5.0–150.0 mg L⁻¹ (r = 0.9953). The RSD value was 2.8% for intra-day precision and 5.4% inter-day precision. The recoveries of the standard addition of tablets and human urine ranged from 90.3% to 107.5% and from 82.0 to 105.7%, respectively. The proposed method was successfully applied to determine FA contents in commercial pharmaceutical tablets and human urine samples. Results suggested that this method was simple and robust.

A capillary electrophoresis method with on-line inhibited chemiluminescence detection was first used to determine folic acid.     

应用高效毛细管电泳技术快速分离尿肌酐

目的建立1种快速测定尿中肌酐浓度的毛细管电泳方法.方法用pH2.5、0.1 mmol/L磷酸作为电泳缓冲液,利用非涂层石英毛细管48.5 cm×50 μm(id),采用外标法定量,检测波长191 nm.结果本法在34.5～8 840 μmol/L浓度范围内呈良好的线性(r=0.998),日内和日间变异系数分别为3.5%、6.2%,平均回收率94.7%.与苦味酸法有良好的相关性(Y=1.004X+0.004,r=0.998,n=45).结论本法线性范围宽、简单、快速,可应用于临床样品的检测.     

毛细管区带电泳对M蛋白的免疫分型

目的 探讨毛细管区带电泳免疫削减法(ISCZE)在M蛋白免疫分型中的应用价值.方法 运用血清免疫固定电泳(IFE)对645例患者血清进行免疫分型,将其阳性结果与ISCZE法免疫分型结果进行比较.结果 ISCZE法出现M蛋白的阳性率为20.3%,IFE为22.6%,分型结果基本一致.结论 对于特征性的、大量的单克隆免疫球蛋白,毛细管区带电泳可有效完成对M蛋白的免疫分型.     

Diagnosis of congenital disorders of glycosylation by capillary zone electrophoresis of serum transferrin

BACKGROUND: Congenital disorders of glycosylation (CDG) are usually diagnosed by isoelectric focusing (IEF) of serum transferrin (Tf). The aim of this study was to evaluate capillary zone electrophoresis (CZE) as a diagnostic alternative to IEF. METHODS: We performed 792 CZE analyses of Tf, using the CEofix(TM)-CDT (carbohydrate-deficient transferrin) assay. Peak identification was based on relative migration times (RMTs) to reduce migration variability. RESULTS: Tf profiles comprised three main groups (A-C). Groups A and B were characterized by one or two dominant tetrasialo-Tf peaks, whereas group C showed a widely variable Tf isoform composition. Group A was composed of four subgroups: a major group with a typical Tf profile (considered as reference group), two minor groups with decreased or moderately increased trisialo-Tf isoform, and a group showing the presence of unknown compounds with RMTs similar to mono- and disialo-Tf. However, these compounds were absent on IEF. Group C contained all profiles from patients with confirmed as well as putative CDG. From the reference group, 99% confidence intervals were calculated for the RMTs of the Tf isoforms, and percentiles representing the Tf isoform distributions were defined. CONCLUSIONS: All patients with abnormal IEF results and confirmed CDG were identified by CZE; thus, this method can be used as a diagnostic alternative to IEF in a manner suitable for automation. Because whole serum is analyzed, it should be kept in mind that CZE profiles can show substances other than Tf.     

Carbohydrate-deficient transferrin isoforms measured by capillary zone electrophoresis for detection of alcohol abuse

BACKGROUND: Measurements of carbohydrate-deficient transferrin (CDT) are used as markers of alcohol abuse. We developed a capillary zone electrophoresis (CZE) method aimed at improving accuracy of CDT testing. METHODS: We studied 111 alcohol abusers with Alcohol Use Disorders Identification Test scores >11 and 50 teetotalers. CZE was performed with a P/ACE 5500, fused-silica capillaries, and a CEofix CDT reagent set. After iron saturation, sera were loaded by low-pressure injection, separated at 28 kV, and monitored at 214 nm. We identified the transferrin isoforms by migration times, treatment with 100 U/L neuraminidase, and immunosubtraction with anti-human transferrin and anti-C-reactive protein antibodies. We compared CZE results with current biological markers of alcohol abuse, including the %CDT turbidimetric immunoassay. RESULTS: Migration times of the isoforms were identical in both populations. Asialotransferrin was missing in teetotalers but present in 92% of alcohol abusers. Disialotransferrin was higher in those who consumed excessive amounts of alcohol, whereas mean trisialotransferrin concentration was not affected by alcohol abuse. At cutoffs to maximize sensitivity and specificity, these values were 0.92 and 1 [mean ROC area (MRa), 0.96; 95% confidence interval (CI), 0.93-0.99] for asialotransferrin; 0.84 and 0.94 for the sum of asialo- + disialotransferrin (MRa, 0.94; 95% CI, 0.91-0.98); 0.79 and 0.94 for disialotransferrin (MRa, 0.89; 95% CI, 0.84-0.94); 0.62 and 0.53 for trisialotransferrin (MRa, 0.58; 95% CI, 0.49-0.68); 0.79 and 0.82 for a 3% %CDT; and 0.83 and 0.69 for a 2.6% cutoff (MRa, 0.87; 95% CI, 0.81-0.92). Current markers lack sensitivity (<0.65). Transferrins were not significantly correlated with serum enzymes and mean erythrocyte volume. CONCLUSIONS: CZE-isolated desialylated transferrin isoforms allowed differentiation between chronic alcohol abusers and teetotalers.