Carne T J, Huber R E, Davitt P, Edwards L A
J Chromatogr. 1986 Oct 3;367(2):393-403. doi: 10.1016/s0021-9673(00)94860-x.
Separation of tyrosine, fluorotyrosine, monoiodotyrosine and diiodotyrosine was achieved by reversed-phase high-performance liquid chromatography (HPLC) using a gradient of acetonitrile with water and using trifluoroacetic acid for ion pairing. No derivatization of the amino acids, prior to separation, was needed. The spectral properties of Tyr and its fluorine and iodine derivatives and the dependence of their absorbance maxima on pH, made it possible to analyze and differentiate between these derivatives in the free amino acid form or in peptides. This analysis was accomplished by adjusting the post column HPLC eluate from two identical runs to different pH values and then comparing the spectra of the peaks from these two runs with a diode array detector. Hydrolysis in 6 M hydrochloric acid was totally destructive to mono- and diiodotyrosine. However, base hydrolysis in 13.5 M sodium hydroxide for 30 min at 121 degrees C in an autoclave caused no destruction and allowed excellent recovery of all of the Tyr derivatives. This is the first report of simple methods for the detection and analysis of these amino acids and of a hydrolytic method which protects against their loss. A method of storage was also proposed.
通过反相高效液相色谱法（HPLC），利用乙腈与水的梯度洗脱，并使用三氟乙酸进行离子配对，实现了酪氨酸、氟酪氨酸、单碘酪氨酸和二碘酪氨酸的分离。分离前无需对氨基酸进行衍生化处理。酪氨酸及其氟化物和碘化物的光谱特性，以及它们的最大吸收波长对pH值的依赖性，使得能够以游离氨基酸形式或肽形式对这些衍生物进行分析和区分。该分析通过将两次相同运行的柱后HPLC洗脱液调节至不同的pH值，然后使用二极管阵列检测器比较这两次运行中峰的光谱来完成。在6 M盐酸中水解对单碘酪氨酸和二碘酪氨酸具有完全破坏性。然而，在高压釜中于121℃用13.5 M氢氧化钠进行30分钟的碱水解不会造成破坏，并且能很好地回收所有酪氨酸衍生物。这是关于这些氨基酸检测和分析的简单方法以及防止其损失的水解方法的首次报道。还提出了一种储存方法。
