Journal of Chromatography, 637 (1993) 91- 95 Elsevier Science Publishers B.V., Amsterdam CHROM. 25 003 Short Communication Determination of the average molecular size of glycosaminoglycans by fast protein liquid chromatography James Melrose* and Peter Ghosh The Ray mond Purves Bone and Joint Laboratories, Universiry of Sydney, Royal North Shore Hospital, St. Leonards, NSW 2065 (Australia) (Received January 19th, 1993) ABSTRACT The determination of the molecular masses of glycosaminoglycans (GAGS) has in the past been hampered by a lack of readily available standards. In the present study methods were determined for the fractionation of commercially available bovine tracheal chondroitin sulphate A into essentially mono-disperse GAG pools which were shown to be suitable as standards for the calibration of Superose 6 and 12 fast protein liquid chromatograph (PPLC) columns. Superose 6 PPLC was particularly suitable for assessing the size distribution of GAGs of M, 10000-40000 and the relatively high flow-rates possible with this support enabled a considerably faster analysis of samples compared to the soft gels previously used for this purpose. INTRODUCTION In 1971 Wasteson [1,2] published details of the fractionation and characterisation of glycosaminoglycans (GAGS) from ox nasal septa using Sephadex G200 gel permeation chromatog- raphy [ 1,2]. Despite the relatively polydisperse nature of the GAGS examined, discrete essen- tially mono-disperse pools were nevertheless prepared which were shown to be suitable as standards for assigning size distributions to un- known GAG samples. However, the soft gel matrices employed which necessitated slow flow- rates precluded the rapid analysis of samples. In the current study, the original methodology of * Corresponding author. Wasteson was employed to prepare chondroitin sulphate standards from bovine tracheal chon- droitin sulphate A, a commercially available source, and these were subsequently used to calibrate Superose 6 and 12 fast protein liquid chromatography (FPLC) columns. The molecu- lar mass values obtained by FPLC were compar- able to those obtained earlier with softer gel matrices [1,2], however a considerably faster analysis of samples was possible (cu. 1 h per sample), since a relatively fast flow-rate was possible with these FPLC matrices. EXPERIMBNTAL Reagents Sephadex G200, Superose 6 and 12 pre-packed HR lo/30 FPLC columns were obtained from 0021-9673/93/$06.00 0 1993 Elsevier Science Publishers B.V. All rights reserved