反相离子对液相色谱法快速分析丙酮醛双（脒腙）

Rapid analysis for methylglyoxal bis(guanylhydrazone) by reversed-phase ion-pair liquid chromatography.

作者信息

Gildersleeve D L, Tobes M C, Natale R B

出版信息

Clin Chem. 1985 Dec;31(12):1979-84.

Abstract

We describe a "high-performance" reversed-phase ion-pair liquid-chromatographic procedure for measuring methylglyoxal bis(guanylhydrazone) (MGBG) in plasma, urine, and bone-marrow leukocytes. Specimens of plasma and bone-marrow leukocytes are deproteinized with perchloric acid, then neutralized with KOH. Urinary MGBG is isolated by liquid-solid extraction in a C18 Sep-Pak. The chromatographic system consists of a 45 X 4.6 mm (i.d.) octadecylsilyl (C18, 5-microns particle) column and a mobile phase consisting of methanol/sodium acetate buffer (200 mmol/L, pH 4.5), 2/3, by vol. The acetate buffer also contains 20 mmol of 1-octanesulfonate and 40 mg of sodium azide per liter. The column effluent is monitored at 283 nm. At a flow rate of 3.0 mL/min, MGBG is eluted in 1.67 min. The detection limit is 20 nmol/L, and peak height varies linearly with concentration from 0.02 to 40 mumol/L. Analytical recovery exceeds 99%. Within-day CVs ranged from 0.9% to 2.9%, between-day CVs from 4.2% to 6.2%.

摘要

我们描述了一种用于测定血浆、尿液和骨髓白细胞中甲基乙二醛双（脒腙）（MGBG）的“高性能”反相离子对液相色谱法。血浆和骨髓白细胞标本用高氯酸脱蛋白，然后用氢氧化钾中和。尿中的MGBG通过在C18 Sep-Pak柱上进行液固萃取分离。色谱系统由一根45×4.6 mm（内径）的十八烷基硅烷（C18，5微米粒径）柱和一种流动相组成，流动相为甲醇/醋酸钠缓冲液（200 mmol/L，pH 4.5），体积比为2/3。醋酸盐缓冲液每升还含有20 mmol的1-辛烷磺酸盐和40 mg的叠氮化钠。在283 nm处监测柱流出物。流速为3.0 mL/min时，MGBG在1.67分钟内洗脱。检测限为20 nmol/L，峰高在0.02至40 μmol/L的浓度范围内与浓度呈线性关系。分析回收率超过99%。日内变异系数范围为0.9%至2.9%，日间变异系数范围为4.2%至6.2%。