•HA, CS, DS and heparin were separated.•A new buffer system was employed.•A fast and steady plasma GAGs extraction method was established.This study reports the use of diethylenetriamine as background electrolyte for the simultaneous separation of hyaluronan acid, chondroitin sulfate, dermatan sulfate and heparin. The analytes were baseline separated by using an uncoated fused silica capillary at 37 °C with a run time of 23 min. The migration order, with hyaluronan acid at first and heparin at last, was related to the sulfation degree. The effect of salt concentration on resolution and migration order was also investigated. The developed method was applied to the simultaneous determination of hyaluronan acid and chondroitin sulfate in mouse plasma. Interferences in plasma were removed by protein precipitation and glycosaminoglycans were further purified by ethanol precipitation. The method was validated over the concentration range from 50 to 600 μg/mL for hyaluronan acid and 500 to 6000 μg/mL for chondroitin sulfate in mouse plasma. Results from assay validations showed that the method was selective and robust.

An electrophoretically mediated microanalysis (EMMA) protocol for the determination of different chondroitin sulfate (CS) origins based on the difference in the content of unsaturated disaccharides produced by degradation with chondroitinase ABC was developed. Separations were performed in an uncoated fused silica capillary (total length: 60.2 cm, effective length: 50 cm, 50 μm i.d.) at 20 kV and 37 °C. The influences of various parameters, such as different kinds of separation buffers, substrate concentration and incubation time, on separation were investigated. The optimum conditions were as follows: separation buffer, 25 mM tetraborate buffer (pH 9.5); incubation buffer, 50 mM Tris-60 mM acetate buffer (pH 8.0); sample injection, 5 s at 0.5 psi; CS concentration, 500 μg mL−1; incubation time, 8 min. Nonsulfated, monosulfated, disulfated and trisulfated Δ-disaccharides were separated well under the above optimal conditions. The developed method was used to determine the amounts of disaccharides in CS from different sources and the results were compared with those obtained by offline analysis. The results indicated that the developed method could successfully distinguish CS with minor differences and could obtain good coherence results compared with the traditional method.

Chondroitin sulfate (CS) is an example of a type of acidic mucopolysaccharides, which consist of repeating disaccharide units of glucuronic acid and galactosamine. Since CS has no UV chromophore it is usually detected by its terminal absorption at a wavelength of 200 nm due to the N-acetyl function, resulting in lower sensitivity. Raman spectroscopy (Raman) and near-infrared spectroscopy (NIR) coupled with partial least squares (PLS) can provide rapid, simple, reproducible and non-destructive quantitative analysis of CS, and no sample pre-treatment or pre-separation are required. In the present study we predicted the CS content of tablets using Raman and NIR combined with PLS. Our results showed that the predicted values obtained by NIR were in good agreement with the actual values, with a correlation coefficient of 0.994. In Raman spectroscopy studies, when the CS content of tablets was within the range 7–39% the correlation coefficient and root mean square error of calibration (RMSEC) were 0.998 and 0.578, respectively. When the CS content of tablets was within the range 41–67% the correlation coefficient and RMSEC were 0.994 and 0.742, respectively. High accuracy could thus be achieved within a wider concentration range by use of NIR, whereas relatively high accuracy could be obtained only within a limited concentration range by the use of Raman spectroscopy.

Chondroitin sulfate (CS) is an example of a type of acidic mucopolysaccharides, which consist of repeating disaccharide units of glucuronic acid and galactosamine. Since CS has no UV chromophore it is usually detected by its terminal absorption at a wavelength of 200 nm due to the N-acetyl function, resulting in lower sensitivity. Raman spectroscopy (Raman) and near-infrared spectroscopy (NIR) coupled with partial least squares (PLS) can provide rapid, simple, reproducible and non-destructive quantitative analysis of CS, and no sample pre-treatment or pre-separation are required. In the present study we predicted the CS content of tablets using Raman and NIR combined with PLS. Our results showed that the predicted values obtained by NIR were in good agreement with the actual values, with a correlation coefficient of 0.994. In Raman spectroscopy studies, when the CS content of tablets was within the range 7–39% the correlation coefficient and root mean square error of calibration (RMSEC) were 0.998 and 0.578, respectively. When the CS content of tablets was within the range 41–67% the correlation coefficient and RMSEC were 0.994 and 0.742, respectively. High accuracy could thus be achieved within a wider concentration range by use of NIR, whereas relatively high accuracy could be obtained only within a limited concentration range by the use of Raman spectroscopy.