Citation: YANG Xiang-Fen, ZHANG Ke, WANG Shi-Cong, YU Juan, NIU Xiao-Ya, CHEN Wei, WU Nian, SONG Qing-Qing, SONG Yue-Lin. Rapid Tryptic Peptide Mapping of Human Hemoglobin Using Direct Infusion Coupled to MS/MS^ALL[J]. Chinese Journal of Analytical Chemistry, ;2023, 51(3): 383-389. doi: 10.19756/j.issn.0253-3820.221384 shu

Rapid Tryptic Peptide Mapping of Human Hemoglobin Using Direct Infusion Coupled to MS/MS^ALL

YANG Xiang-Fen ¹ ,
ZHANG Ke ¹ ,
WANG Shi-Cong ² ,
YU Juan ² ,
NIU Xiao-Ya ¹ ,
CHEN Wei ¹ ,
WU Nian ¹ ,
SONG Qing-Qing ^1,, ,
SONG Yue-Lin ^1,,

1.
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China;
2.
Zhangzhou Pien-Tze-Huang Pharmaceutical Co., Ltd., Zhangzhou 363000, China

Corresponding author: SONG Qing-Qing, SONG Yue-Lin,
Received Date: 28 July 2022
Revised Date: 4 November 2022

Fund Project: Supported by the National Natural Science Foundation of China (Nos. 81973444, 82003911).

Peptide mapping currently serves as the primary means for quality control of protonic drugs through employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) as the workhorse. Although being versatile, LC-MS/MS still suffers from shortcomings in terms of time-consuming, solvent-intensive and sophisticated instrumentation. Because of the extraordinary merit namely high throughput, the potential of direct infusion (DI) coupled to tandem mass spectrometry towards the tryptic peptide mapping was evaluated, and particularly, a new technique termed as MS/MS^ALL program was implemented to acquire MS² spectrum for each nominal m/z value, attributing to the involvement of the robust gas phase ion fractionation technique. Human hemoglobin (Hb) was utilized as a proof-of-concept target molecule. After thorough tryptic digestion, the peptide pool was directly injected into the ion source of mass spectrometer through an infusion pump, and MS/MS^ALL was programmed to record all desired MS² spectra, because the whole ion population was fractionated into sequential ion currents with 1 Da mass window. After matching the acquired MS¹ and MS² spectral information, 20 over 21 theoretical peptides that were suggested by Skyline software, were successfully captured, whilst all 21 peptides were detected by the well-defined LC-MS/MS program. Moreover, similar MS¹ spectra (average spectrum for LC-MS/MS) occurred between the two means, primarily including quasi-molecular ions bearing the charged states amongst 1-4. Either analytical method dominantly gave out characteristic y⁺ ions. Thereafter, comparable potential towards tryptic peptide mapping was yielded from DI-MS/MSALL in comparison with LC-MS/MS, and the DI-MS/MSALL exhibited many advantages including high throughput, solvent-saving and low-costing measurement. The results demonstrated that the DI-MS/MSALL could be an alternative analytical tool for LC-MS/MS.
- Tryptic peptide mapping,
- Direct infusion-MS/MS^ALL,
- Human hemoglobin,
- Proteinic drugs
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Rapid Tryptic Peptide Mapping of Human Hemoglobin Using Direct Infusion Coupled to MS/MSALL

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Rapid Tryptic Peptide Mapping of Human Hemoglobin Using Direct Infusion Coupled to MS/MS^ALL