Yu T J, Schwartz H A, Cohen S A, Vouros P, Karger B L
J Chromatogr. 1984 Oct 5;301(2):425-40. doi: 10.1016/s0021-9673(01)89216-5.
N-Acetyl-N,O,S-permethylated derivatives of oligopeptides were analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS) using a moving belt interface. A heated-gas nebulizer was employed for sample deposition, thus permitting the effective use of a water-methanol gradient covering the range from 5% to 95% water at mobile phase flow-rates of 0.5 ml/min. We demonstrate in this paper that it is possible to sequence the octapeptide derived from the C-chain of glucagon by HPLC-MS analysis of a permethylated enzymatic hydrolysate of this peptide using overlap information from the mass spectral patterns. Moreover, it is shown that peptides not readily amenable to analysis by gas chromatography-MS can be analyzed using this approach. Preliminary results suggest that N-acetyl-methyl ester derivatives of oligopeptides may in specific cases also be a useful alternative for HPLC-MS analysis of complex oligopeptide mixtures.
使用移动带接口的高效液相色谱-质谱联用仪（HPLC-MS）对寡肽的N-乙酰基-N、O、S-全甲基化衍生物进行了分析。采用加热气体雾化器进行样品沉积，从而能够在0.5 ml/min的流动相流速下有效使用覆盖5%至95%水范围的水-甲醇梯度。我们在本文中证明，通过对该肽的全甲基化酶解产物进行HPLC-MS分析，并利用质谱图中的重叠信息，可以对源自胰高血糖素C链的八肽进行测序。此外，结果表明，不易用气相色谱-质谱联用仪分析的肽也可以用这种方法进行分析。初步结果表明，在特定情况下，寡肽的N-乙酰基甲酯衍生物也可能是用于复杂寡肽混合物HPLC-MS分析的有用替代物。
