Burke R A, Shockley T R
Baxter Healthcare, Renal Division, McGaw Park, Il 60085, USA.
J Chromatogr B Biomed Sci Appl. 1998 Oct 23;718(1):115-20. doi: 10.1016/s0378-4347(98)00363-6.
A method using gel permeation chromatography was evaluated for the quantitation of dextran 70 in dialysate samples containing polyglucose. Dialysate samples containing dextran 70 and polyglucose were pretreated using the enzyme alpha-amylase to selectively hydrolyze the alpha(1-4)-linked polyglucose, while leaving the alpha(1-6)-linked dextran 70 intact. Following sample deproteinization with trichloroacetic acid, dextran 70 was quantitated using gel permeation chromatography with refractive index detection. This method was evaluated for accuracy, precision, specificity, linearity, range, and analyte stability. Adequate method linearity with a correlation of >0.999 was established over the range of dextran 70 concentration from 1 to 0.025 mg/ml. Method precision was approximately 2% R.S.D. and accuracy (% recovery) was approximately 98-100% in the typical sample concentration range (1-0.5 mg/ml). This method was applied to the determination of intraperitoneal fluid kinetics in continuous ambulatory peritoneal dialysis (CAPD) patients administered daily night-time intraperitoneal exchanges with either 7.5% polyglucose or 4.25% dextrose. Dextran 70 was added to the dialysis solutions to yield an initial concentration of 1 mg/ml. Dialysate samples were collected at various times over a 10-h dwell-time and assayed for dextran 70. Intraperitoneal volume profiles based on dextran 70 concentrations and drain volumes were then calculated for each dialysis solution.
评估了一种使用凝胶渗透色谱法对含有聚葡萄糖的透析液样品中的右旋糖酐70进行定量的方法。含有右旋糖酐70和聚葡萄糖的透析液样品用α-淀粉酶进行预处理,以选择性地水解α(1-4)连接的聚葡萄糖,同时使α(1-6)连接的右旋糖酐70保持完整。用三氯乙酸对样品进行脱蛋白处理后,使用带有折光指数检测的凝胶渗透色谱法对右旋糖酐70进行定量。对该方法的准确性、精密度、特异性、线性、范围和分析物稳定性进行了评估。在1至0.025 mg/ml的右旋糖酐70浓度范围内建立了足够的方法线性,相关性>0.999。在典型样品浓度范围(1至0.5 mg/ml)内,方法精密度约为2%相对标准偏差,准确度(回收率%)约为98 - 100%。该方法应用于对接受每日夜间腹腔内交换7.5%聚葡萄糖或4.25%葡萄糖的持续性非卧床腹膜透析(CAPD)患者的腹腔液动力学进行测定。向透析液中加入右旋糖酐70,使其初始浓度为1 mg/ml。在10小时的驻留时间内的不同时间收集透析液样品,并测定其中的右旋糖酐70。然后根据每种透析液的右旋糖酐70浓度和排出体积计算腹腔容积曲线。