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Determination of amino acids in polymers with varying structure and charge was performed using vapor phase acid hydrolysis and subsequent precolumn derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Percent load of neutral, cationic and anionic peptide-modified synthetic polymers was accurately determined using this technique. Assay utility was shown for glycosaminoglycans and other sulfated polymers, neutral carbohydrate polymers such as agar, agarose, and cellulose, and polymers such as lipopolysaccharide and deoxyribonucleic acid. The carboxylated and sulfated molecules included chondroitin sulfate, hyaluronic acid, dermatan sulfate, and heparin, and the sulfated polymers included fucoidan, carrageenan, and dextran sulfate, as examples. Assayed cumulative amino acid concentrations (i.e. protein levels) are reported, although amino acid distribution data was also available from the analysis. Recovery was acceptable for the various compounds tested and did not correlate with structure. However, different sample sizes were necessary to achieve acceptable recovery, depending on the level of protein present in the matrix. While some matrices contained peaks in addition to the amino acids and amino sugars, they were not found to interfere using the standard gradient separation. Assayed amino acid profiles were compared for agaroses with differing electroendosmosis values and for agar samples from different parts of the globe. While the amounts of protein varied depending on source, the relative distribution of amino acids was very similar across the agar samples surveyed.

Low-level (subanomole) determination of amino acids in samples of naturally derived polymeric carbohydrates has been demonstrated using vapor-phase acid hydrolysis and subsequent precolumn derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Application has been demonstrated for neutral polysaccharide polymers such as laminarin (beta-1,3; branched), curdland (beta-1,3; unbranched), pullulan (alpha-1, 6-maltotriose), glycogen (alpha-1,4-glucan), and inulin (polyfructose). Successful determination (acceptable recovery and lack of interferences) was possible in samples which also contained up to roughly 50 micrograms amino sugars (e.g., chitosan or glucans with copurified glucosamine oligomers), although optimum utility is for samples containing up to ca. 10 micrograms total amines. The limit of quantification was roughly 20 ng protein/ mg sample, based on analysis of reagent blanks, although the limit of detection was much lower (ca. 0.1 ng protein/mg sample). Incorporation of a relatively rapid hydrolysis (150 degrees C for 1.5 h) gave similar results to use of 110 degrees C for 24 h and allowed for relatively rapid processing. The method has shown good sensitivity, linearity, ruggedness, and ease. Recovery has been optimized, although yield varied somewhat depending on polymer composition.