Palace G P, Fitzpatrick R, Tran K V, Phoebe C H, Norton K
Analytical Chemistry and Chemistry Departments, GelTex Pharmaceuticals, Incorporated, Waltham, MA 02451, USA.
Biochim Biophys Acta. 1999 Nov 16;1472(3):509-18. doi: 10.1016/s0304-4165(99)00154-3.
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.
采用气相酸水解法及随后用6-氨基喹啉-N-羟基琥珀酰亚胺基甲酸酯（AQC）进行柱前衍生化，对结构和电荷各异的聚合物中的氨基酸进行测定。使用该技术可准确测定中性、阳离子和阴离子肽修饰的合成聚合物的负载百分比。该分析方法对糖胺聚糖和其他硫酸化聚合物、中性碳水化合物聚合物（如琼脂、琼脂糖和纤维素）以及聚合物（如脂多糖和脱氧核糖核酸）均有效。羧化和硫酸化分子包括硫酸软骨素、透明质酸、硫酸皮肤素和肝素，硫酸化聚合物包括岩藻依聚糖、卡拉胶和硫酸葡聚糖等。尽管分析中也可获得氨基酸分布数据，但报告的是测定的累积氨基酸浓度（即蛋白质水平）。对所测试的各种化合物而言，回收率是可接受的，且与结构无关。然而，根据基质中蛋白质的含量，需要不同的样品量才能实现可接受的回收率。虽然有些基质除了氨基酸和氨基糖外还含有峰，但使用标准梯度分离法未发现它们有干扰。比较了具有不同电渗值的琼脂糖以及来自全球不同地区的琼脂样品的测定氨基酸谱。虽然蛋白质含量因来源而异，但在所调查的琼脂样品中，氨基酸的相对分布非常相似。
