Ackermann B L, Coutant J E, Chen T M
Marion Merrell Dow R&D, Cincinnati, Ohio 45215-6300.
Biol Mass Spectrom. 1991 Jul;20(7):431-40. doi: 10.1002/bms.1200200707.
The ability to acquire structurally informative daughter ion spectra for individual peptides undergoing separation and analysis by continuous flow fast atom bombardment (CF FAB) is demonstrated. To illustrate the potential of this methodology, tryptic and chymotryptic digests of the 29-residue peptide glucagon were analyzed by CF FAB using mass spectrometric and tandem mass spectrometric detection in consecutive analyses. Daughter ion spectra were recorded using B/E linked scans for the major hydrolysis products observed by liquid chromatography/mass spectrometry. The peptide mixtures were separated by gradient capillary high-performance liquid chromatography with the FAB matrix being added post-column using a coaxial flow interface between the column and flow probe. The entire effluent (3 microl min(-1)) was sampled by the mass spectrometer. Results obtained using less than 300 pmol of digested glucagon indicated several advantages to tandem mass spectrometric detection including the ability to confirm identities for products of enzymatic digestion and the potential use of this method for tandem sequence analysis of peptide mixtures.
本文展示了通过连续流快速原子轰击（CF FAB）对正在进行分离和分析的单个肽段获取具有结构信息的子离子谱的能力。为了说明这种方法的潜力，使用CF FAB对由29个氨基酸残基组成的胰高血糖素肽进行胰蛋白酶和胰凝乳蛋白酶消化，并在连续分析中采用质谱和串联质谱检测。使用B/E联动扫描记录通过液相色谱/质谱观察到的主要水解产物的子离子谱。肽混合物通过梯度毛细管高效液相色谱进行分离，FAB基质在柱后使用柱与流动探头之间的同轴流动接口添加。质谱仪对全部流出物（3 μl min⁻¹）进行采样。使用少于300 pmol的胰高血糖素消化产物所获得的结果表明，串联质谱检测具有几个优点，包括能够确认酶消化产物的身份以及该方法在肽混合物串联序列分析中的潜在用途。
