Friman C, Juvani M
Scand J Rheumatol. 1977;6(2):87-91. doi: 10.3109/03009747709095426.
Plasma glycosaminoglycans (GAG) were isolated from 10 ml of plasma by a modification of the method of Calatroni et al. (3). DE-52 anion-exchange cellulose was used in the isolation of the fraction operationally defined as free GAG. Chondroitin sulphate and heparin added to plasma were quantitatively recovered in this fraction. After proteolysis with papain the fraction operationally defined as bound GAG was isolated using anion-exchange resin AG 1 X 2. GAG were measured as hexuronate with the m-hydroxydiphenyl method of Blumenkrantz and Asboe-Hansen (7) which was superior to various modifications of the carbazole/borate carbazole procedures. In 15 healthy females and in 15 healthy males the concentrations of the free GAG (mean +/- S.D., expressed as microgram per 10 ml of plasma) were: 12.2 +/- 2.8 and 16.8 +/- 3.8 (P less than 0.001); of the bound GAG 40.4 +/- 7.7, and 40.2 +/- 11.6; and of the total GAG 52.7 +/- 9.0 and 57.0 +/- 10.4, respectively. With the isolation procedures used, plasma GAG were obtained in sufficient quantity for their electrophoretic characterization. Assay of plasma GAG can be performed with satisfactory accuracy and precision within two days by the present method. In clinical chemistry its application to the study of proteoglycan and GAG metabolism in various diseases may prove valuable.
采用对卡拉特罗尼等人(3)的方法进行改良的方式,从10毫升血浆中分离血浆糖胺聚糖(GAG)。在分离操作上定义为游离GAG的组分时使用DE - 52阴离子交换纤维素。添加到血浆中的硫酸软骨素和肝素在该组分中被定量回收。用木瓜蛋白酶进行蛋白水解后,使用阴离子交换树脂AG 1 X 2分离操作上定义为结合GAG的组分。采用布卢门克兰茨和阿斯博 - 汉森(7)的间羟基联苯法将GAG测定为己糖醛酸,该方法优于咔唑/硼酸盐咔唑法的各种改良方法。15名健康女性和15名健康男性的游离GAG浓度(均值±标准差,以每10毫升血浆中的微克数表示)分别为:12.2±2.8和16.8±3.8(P<0.001);结合GAG浓度分别为40.4±7.7和40.2±11.6;总GAG浓度分别为52.7±9.0和57.0±10.4。采用所使用的分离程序,可获得足够量的血浆GAG用于其电泳表征。通过本方法可在两天内以令人满意的准确度和精密度进行血浆GAG的测定。在临床化学中,其在各种疾病中蛋白聚糖和GAG代谢研究中的应用可能具有重要价值。