PubMed · 13619780
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M SCHUBERT. 1958. Chondromucoprotein.. https://pubmed.ncbi.nlm.nih.gov/13619780/
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A novel analytical method for determination of total amount of chondroitin sulfate (CS) based on its conversion to desulfated chondro-disaccharide via an enzyme-catalyzed reaction, was developed. Using the in-capillary enzyme reaction, the method was also applied to the successful construction of an on-line analytical system. Within this system, electrophoretic migration was used to mix zones containing the enzyme mixture (chondroitinase ABC, chondro-4-sulfatase, chondro-6-sulfatase and 2-o-sulfatase) and the substrate (CS). The reaction was then allowed to proceed in the presence of a weak electric field and, finally, the product (desulfated chondro-disaccharide) of enzyme reaction migrated to the detector under the influence of an applied electric field. A polyvinyl alcohol-coated capillary was used to reduce protein adsorption. Desulfated chondro-disaccharide was successfully migrated toward the anode in 10 mM Tris-acetate buffer (pH 7.0) under reversed polarity and detected at 232 nm. The established method was validated and demonstrated to be applicable in the determination of total amount of CS in a commercial ophthalmic solution. No interference from the formulation excipients was observed. Good linearity was obtained, with correlation coefficients above 0.999. Recoveries and precisions ranged from 100.0 to 100.5%, and from 0.2 to 0.6% of the relative standard deviation, respectively. Good agreement was obtained between the established method and traditional photometric method based on carbazole reaction. In this study, application of the method to disaccharide compositional analysis was also performed.
The group index of the cornea, rather than the phase refractive index, is required for thickness calculations with optical coherence tomography. Recent advances with high-speed optical coherence tomography at 1.3 microm make index measurement at this wavelength of great interest. Group indices of three human corneas from an eye bank were measured in vitro with optical coherence domain reflectometry. Measurements were made in a calibrated cuvette filled with a preservation medium to maintain proper corneal hydration. Group indices were calculated from the optical path lengths measured. The corneal group index was 1.389 +/- 0.004 (average +/- standard deviation). The average group index of a balanced salt solution, an approximation to aqueous humor, was 1.343 +/- 0.001.
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