Monitoring of solid phase peptide synthesis by an automated spectrophotometric picrate method.
Monitoring of solid phase peptide synthesis by an automated spectrophotometric picrate method.
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通过自动分光光度苦味酸盐方法监测固相肽合成。
DOI：
10.1016/0003-2697(75)90509-6
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发表时间：
1975
影响因子：
2.9
通讯作者：
R. B. Merrifield
中科院分区：
文献类型：
--
作者：
R. Hodges;R. B. Merrifield
构建了一种自动监测固相肽合成过程的仪器。其测量Nα保护基脱保护后的总肽链以及偶联反应后未反应肽链的数量。它是基于分光光度苦味酸盐法测定树脂结合的氨基。对于不含组氨酸的肽，游离氨基用苦味酸在二氯甲烷中转化为苦味酸盐，而含组氨酸的肽用组氨酸上的Nim-DNP保护基合成，游离氨基用咪唑鎓苦味酸盐转化为苦味酸盐。用二异丙基乙胺置换苦味酸盐，在362 nm处定量测定其吸光度。该方法的范围为约0.4-4000 μ mol的氨基。对于含组氨酸的肽，偶联的估计值仅约为± 1%，但对于不含组氨酸的肽，偶联的估计值约为± 0.2%。通过与Schiff碱法和色谱法的比较，对该方法的准确度进行了评价，并将其应用于几种小肽的合成。
An instrument was constructed which automatically monitors the progress of solid phase peptide syntheses. It measures total peptide chains, after deprotection of the Nαprotecting group, and the number of unreacted peptide chains after the coupling reaction. It is based on the spectrophotometric picrate method for the determination of resin-bound amino groups. For peptides not containing histidine the free amino groups are converted to picrate salts with picric acid in dichloromethane, while peptides containing histidine are synthesized with Nim-DNP protecting groups on histidine and the free amino groups are converted to picrate salts with imidazolium picrate. The picrate is displaced by diisopropylethylamine, and its absorbance is measured quantitatively at 362 nm. The range of the method was approximately 0.4–4000 μmoles of amino groups. With histidine-containing peptides the estimate of coupling was only about ±1%, but with peptides not containing histidine it was approximately ±0.2%. The method was evaluated for accuracy by comparison with Schiff base and chromatographic methods and was applied to the synthesis of several small peptides.