Adamson J G, Hoang T, Crivici A, Lajoie G A
Guelph-Waterloo Centre for Graduate Work in Chemistry, Department of Chemistry, University of Waterloo, Ontario, Canada.
Anal Biochem. 1992 Apr;202(1):210-4. doi: 10.1016/0003-2697(92)90229-z.
A chromatographic assay has been developed to quantitate racemization occurring during attachment of protected amino acids to peptide synthesis resins. Acidolytic cleavage of deprotected amino acids from supports and subsequent derivatization with 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide (Marfey's reagent) gave diastereomers separable by reverse-phase HPLC using aqueous acetonitrile. The assay is reliable to 0.1% racemate with a detection limit of approximately 100 pmol. The technique was applied to several acid-labile resins and was used to evaluate the racemizing characteristics of various anchoring methods.
已开发出一种色谱分析方法，用于定量在将受保护氨基酸连接到肽合成树脂过程中发生的消旋化。从载体上酸解切割去保护的氨基酸，随后用1-氟-2,4-二硝基苯基-5-L-丙氨酸酰胺（马尔费试剂）进行衍生化，得到的非对映异构体可用反相高效液相色谱法在乙腈水溶液中分离。该分析方法对消旋体的检测可靠度为0.1%，检测限约为100皮摩尔。该技术应用于几种酸不稳定树脂，并用于评估各种锚定方法的消旋化特性。
