McKay D B, Tanner G P, Maclean D J, Scott K J
Department of Biochemistry, University of Queensland, St. Lucia, Australia.
Anal Biochem. 1987 Sep;165(2):392-8. doi: 10.1016/0003-2697(87)90287-9.
A new and more sensitive cuprammonium reagent containing strong base has been developed for the colorimetric detection of polyols and sugars. A reagent blank with an absorbance at 280 nm, less than half that of previous methods, was achieved by reducing the cupric ion and ammonia concentrations to 1 mM (from 5 mM) and 0.4 M (from 2.25 M), respectively, and adding 0.2 M sodium hydroxide. Polyols are particularly well suited to the new method, showing up to an eightfold increase in sensitivity. The new reagent is readily applicable to the postcolumn detection of polyols and sugars after high-performance liquid chromatography and is linear over the range 40-180 nmol for D-glucitol (sorbitol) and 10-300 nmol for glucose, using standard HPLC equipment.
一种新型且更灵敏的含强碱的铜氨试剂已被开发出来,用于比色检测多元醇和糖类。通过将铜离子浓度从5 mM降至1 mM,氨浓度从2.25 M降至0.4 M,并添加0.2 M氢氧化钠,获得了在280 nm处吸光度比以前方法低一半以上的试剂空白。多元醇特别适合这种新方法,灵敏度提高了八倍。这种新试剂很容易应用于高效液相色谱后多元醇和糖类的柱后检测,使用标准的高效液相色谱设备,对于D-葡萄糖醇(山梨醇)在40 - 180 nmol范围内呈线性,对于葡萄糖在10 - 300 nmol范围内呈线性。