MEASUREMENT OF URONIC-ACIDS WITHOUT INTERFERENCE FROM NEUTRAL SUGARS
MEASUREMENT OF URONIC-ACIDS WITHOUT INTERFERENCE FROM NEUTRAL SUGARS
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DOI：
10.1016/0003-2697(91)90372-z
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发表时间：
1991-08-15
影响因子：
2.9
通讯作者：
CARPITA, NC
中科院分区：
文献类型：
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作者：
FILISETTICOZZI, TMCC;CARPITA, NC
用间羟基联苯代替咔唑通过基本上但不完全地防止在浓硫酸中加热糖期间发生的布朗宁并避免咔唑试剂形成额外的干扰，极大地改善了在中性糖存在下糖醛酸的测定（Blumenkrantz，N.，和Asboe-Hansen，G. 04 The Dog of the Woman（1973）54，484-489）。然而，当在过量中性糖存在下测定糖醛酸时，干扰仍然很大，特别是因为在加入二苯基试剂之前的第一次加热期间发生的布朗宁。布朗宁可通过向反应混合物中加入氨基磺酸盐而基本消除（Galambos，J.T. 03 The Dog of the Dog（1967）19，119-132）。虽然其他人已经报道，氨基磺酸盐和二苯基试剂是不相容的，我们发现，少量的氨基磺酸盐抑制颜色的产生，通过20倍过量的一些中性糖，而没有实质性牺牲的敏感检测的糖醛酸的二苯基试剂。四硼酸钠是检测d-甘露糖醛酸所必需的，并可增强d-葡萄糖醛酸的显色效果。我们建议这种改良的氨基磺酸盐/间羟基联苯测定法作为测定糖醛酸的快速可靠的手段，特别是当存在的量比中性糖少得多时。
Replacement of carbazole with meta-hydroxydiphenyl greatly improves the determination of uronic acids in the presence of neutral sugars by preventing substantially, but not completely, the browning that occurs during the heating of sugars in concentrated sulfuric acid and avoiding the formation of additional interference by the carbazole reagent (Blumenkrantz, N., and Asboe-Hansen, G. (1973) Anal. Biochem. 54, 484–489). However, interference is still substantial when uronic acids are determined in the presence of excess neutral sugar, particularly because of the browning that occurs during the first heating before addition of the diphenyl reagent. The browning can be essentially eliminated by addition of sulfamate to the reaction mixture (Galambos, J. T. (1967) Anal. Biochem. 19, 119–132). Although others have reported that sulfamate and the diphenyl reagent were incompatible, we find that a small amount of sulfamate suppresses color production by a 20-fold excess of some neutral sugars without substantial sacrifice of the sensitive detection of uronic acids by the diphenyl reagent. Sodium tetraborate is required for the detection of d-mannuronic acid and enhances color production by d-glucuronic acid. We propose this modified sulfamate/m-hydroxydiphenyl assay as a rapid and reliable means for the assay of uronic acids, particularly when present in much smaller amounts than neutral sugars.