Klompmakers A A, Hendriks T
Anal Biochem. 1986 Feb 15;153(1):80-4. doi: 10.1016/0003-2697(86)90064-3.
The absorption spectrum of the dye 1,9-dimethylmethylene blue shifts if complexed with sulfated glycosaminoglycans. The present method uses the decrease in A633 rather than the increase in A535, described in a recent method, to measure the sulfated glycosaminoglycan content of biological samples. A conventional spectrophotometer was used to estimate the levels of sulfated glycosaminoglycan in papain extracts from intestinal wall tissue, by measuring both the A535 and the A633 and comparing them with a chondroitin sulfate standard: a highly significant correlation (r = 0.974, n = 17) was obtained. Also, interference by substances like RNA, DNA, and hyaluronic acid was similar for both methods. These results allowed us to employ a laser densitometer with a helium/neon laser emitting at 633 nm to improve the sensitivity and the capacity of the assay. The combination of a small reaction volume and a high-intensity light source allows the detection of less than 0.1 microgram chondroitin sulfate, a 40-fold improvement in sensitivity as compared with the original method. A very significant correlation (r = 0.885, n = 17) existed between results obtained with the macroassay, using a spectrophotometer, and those found by employing the microassay, using the laser densitometer. The use of microtiter plates and the screening potential of the densitometer yields an assay which is fast, very sensitive, and suitable for processing large numbers of samples.
染料1,9 - 二甲基亚甲蓝与硫酸化糖胺聚糖络合时，其吸收光谱会发生偏移。本方法利用A633的降低而非最近一种方法中所述的A535的增加来测量生物样品中硫酸化糖胺聚糖的含量。使用传统分光光度计通过测量A535和A633并与硫酸软骨素标准品进行比较，来估计肠壁组织木瓜蛋白酶提取物中硫酸化糖胺聚糖的水平：得到了高度显著的相关性（r = 0.974，n = 17）。此外，两种方法中RNA、DNA和透明质酸等物质的干扰情况相似。这些结果使我们能够使用发射波长为633 nm的氦氖激光的激光密度计来提高检测的灵敏度和检测能力。小反应体积与高强度光源的结合使得能够检测到低于0.1微克的硫酸软骨素，与原方法相比灵敏度提高了40倍。使用分光光度计进行的宏观检测结果与使用激光密度计进行的微观检测结果之间存在非常显著的相关性（r = 0.885，n = 17）。使用微量滴定板和密度计的筛选潜力产生了一种快速、非常灵敏且适合处理大量样品的检测方法。
