• Bulletin of the Korean Chemical Society
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• v.26 no.5
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• pp.763-768
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• 2005

An improved tandem time-of-flight (TOF) mass spectrometer for the photodissociation (PD) study of ions generated by matrix-assisted laser desorption ionization (MALDI), MALDI-TOF-PD-TOF, has been designed and constructed. Recording a full spectrum with better than unit mass resolution even in low mass range has been achieved without reflectron voltage stepping which was needed in the previous version. Other aspects of the improvement, such as those in the data system which now allow 10-100 times faster spectral acquisition than with the previous instrument, are described. Rationale for the ideas which have led to the improvements is presented also.

• Nuclear Engineering and Technology
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• v.54 no.2
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• pp.437-441
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• 2022

In this study, we developed a neutron time-of-flight (nTOF) measurement system for a 1.7-MV tandem proton accelerator with a target covered with 300-nm-thick lithium (Li) layer. With implementation of beam chopping module after its ion source, the accelerator is configured to operate in pulsed-beam mode with a pulse width <50 ns at 20-kHz repetition rate. This enables the gamma flash-type nTOF measurement system to identify the neutron generated with 3-MeV proton beam energy. The nTOF system consists of a 30" cylindrical NaI(Tl) and four stilbene scintillation detectors. The NaI(Tl) scintillator is placed 50 cm from the Li target to measure the time of beam irradiation on the target, and the stilbene detectors are placed 2 and 2.4 m away to measure nTOF at each location. The nTOF system successfully measured the generated neutron energy at irradiated proton energies of 2.6 and 3.0 MeV with an average energy resolution of 15%.

• Bulletin of the Korean Chemical Society
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• v.31 no.1
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• pp.92-99
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• 2010

Photoexcitation of a precursor ion inside a cell floated at high voltage installed in a tandem time-of-flight (TOF) mass spectrometer provides triple tandem mass spectrometric information and allows kinetic and mechanistic studies. In this work, the factors affecting, or downgrading, the performance of the technique were identified. Ion-optical and computational analyses showed that an optimum instrument could be designed by utilizing a reflectron with linear-plus-quadratic potential inside. Theoretical predictions were confirmed by tests with instruments built with different ion-optical layout. With optimized instruments, masses of intermediate ions in the consecutive dissociation of a precursor ion could be determined with the maximum error of $\pm5$ Da. We also observed excellent agreement in dynamical parameters (critical energy and entropy) for the dissociation of a model peptide ion determined by instruments with different ion-optical layout operated under optimum conditions. This suggests that these parameters can be determined reliably by the kinetic method developed previously when properly designed and operated tandem TOF instruments are used.

• Bulletin of the Korean Chemical Society
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• v.25 no.12
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• pp.1791-1800
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• 2004

Angiotensin I, a model decapeptide, was glycosylated and partially hydrolyzed with HCl (6 N, 80 $^{\circ}C$, 4 h), aminopeptidase, and carboxypeptidase Y. A single peptide mass map obtained from truncated peptides in the partial acid hydrolysate of angiotensin and its glycosylation product mixture by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry enabled sequencing of angiotensin by a combinatorial procedure. MALDI-TOF and electrospray ionization (ESI) tandem mass spectrometric results indicate that both the N-terminal amino group of aspartic acid and the guanidinium group of the second residue arginine are glycosylated.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.6 no.2
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• pp.134-143
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• 2005

Collagen is the important structural proteins in mammals with various peptide composition and cross-linkings. The direct analysis of collagen protein was not suitable because of its structural complexity and diversity. In this study, we suggest the simple way of collagen analysis by introducing matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS) to identify the collagen and its trypsin-digested fragments, and by subsequent time-of-flight tandem mass spectrometry(Q-TOF MS/MS) to analyze the amino acid sequences of identified fragments. Using the collagen samples extracted from the tail of mouse, 10 separated bands were found in SDS-PAGE, and the masses of most bands could be more finely determined by MALDI-TOF MS. When each 10 separated proteins was tryptic digested and introduced to MALDI-TOF, the Gly1056-Arg1073 fragment from $\alpha_1$-chain was identified in four bands, and the Gly1056-Arg1073 fragment from $\alpha_2$-chain was identified in five bands, both in type I collagen. Although few fragments were found because of the cross-linkings left in digested collagen sample, it could be determined that the type I collagen existed at least in 7 separated bands. When the amino acid sequences of two identified fragments were analyzed by Q-TOF MS/MS, both sequences were identical with those determined by MALDI-TOF MS. It suggested that the two peaks in MALDI-TOF MS caused by the fragments identified in this work could be used as the fingerprint to simply identify type I collagen in protein samples.

• Applied Chemistry for Engineering
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• v.28 no.3
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• pp.263-271
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• 2017

The application of mass spectrometry to polymer science has rapidly increased since the development of MALDI-TOF MS. This review summarizes current polymer analysis methods using MALDI-TOF MS, which has been extensively applied to analyze the average molecular weight of biopolymers and synthetic polymers. Polymer sequences have also been analyzed to reveal the structures and composition of monomers. In addition, the analysis of unknown end-groups and the determination of polymer concentrations are very important applications. Hyphenated techniques using MALDI-tandem MS have been used for the analysis of fragmentation patterns and end-groups, and also the combination of SEC and MALDI-TOF MS techniques is recommended for the analysis of complex polymers. Moreover, MALDI-TOF MS has been utilized for the observation of polymer degradation. Ion mobility MS, TOF-SIMS, and MALDI-TOF-imaging are also emerging technologies for polymer characterization because of their ability to automatically fractionate and localize polymer samples. The determination of polymer characteristics and their relation to the material properties is one of the most important demands for polymer scientists; the development of software and instrument for higher molecular mass range (> 100 kD) will increase the applications of MALDI-TOF MS for polymer scientists.

• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.952-959
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• 2007

His-His-Leu (HHL), a tripeptide derived from a Korean soybean paste, is an angiotensin-I-converting enzyme (ACE) inhibitor. We report here a method of producing this tripeptide efficiently by expressing tandem multimers of the codons encoding the peptide in E. coli and purifying the HHL after hydrolysis of the peptide multimers. The HHL gene, tandemly multimerized to a 40-mer, was ligated with ubiquitin as a fusion gene (UH40). UH40 was inserted into vector pET29b; the UH40 fusion protein was then produced in E. coli BL21. The recombinant UH40 protein was purified by cation-exchange chromatography with a yield of 17.3mg/l and analyzed by matrixassisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry and protein N-terminal sequencing. Leucine aminopeptidase was used to cleave a 405-Da HHL monomer from the UH40 fusion protein and the peptide was purified using reverse-phase high-performance liquid chromatography (HPLC) on a C18 HPLC column, with a final yield of 6.2mg/l. The resulting peptide was confirmed to be HHL with the aid of MALDI-TOF mass spectrometry, glutamine-TOF mass spectrometry, N-terminal sequencing, and measurement of ACE inhibiting activity. These results suggest that our production method is useful for obtaining a large quantity of recombinant HHL for functional antihypertensive peptide studies.

• KSBB Journal
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• v.31 no.4
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• pp.263-269
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• 2016

For decades, the microorganisms in arctic soils have been newly discovered according to the climate change and global warming. In this study, the chemical structure of a lipid A molecule from Pseudomonas sp. strain PAMC 28615 which was newly discovered from arctic soils was characterized by mass spectrometric approaches such as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and MALDI multi-stage tandem mass spectrometry (MS). First, lipopolysaccharide (LPS) from Pseudomonas sp. strain PAMC 28615 was extracted and subsequently hydrolyzed to obtain the lipid A. The parent ion peak at m/z 1632 was determined by MALDI-TOF MS, which also can validate our lipid A purification method. For detailed structural determination, we performed the multiple-stage tandem mass analysis ($MS^4$) of the parent ion, and subsequently the abundant fragment ions in each MS stage are tested. The fragment ions in each MS stage were produced from the loss of phosphate groups and fatty acyl groups, which could be used to confirm the composition or the position of the lipid A components. Consequently, the mass spectrometry-based lipid A profiling method could provide the detail chemical structure of lipid A from the Pseudomonas sp. strain PAMC 28615 as an arctic bacterium from the frozen arctic soil.

• Mass Spectrometry Letters
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• v.7 no.2
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• pp.45-49
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• 2016

An UHPLC-ESI-qTOF-MS analytical method was developed for cyclopeptide alkaloids in the seeds of Ziziphus jujuba var. spinosa (Semen Ziziphi Spinosae), which is a commonly used herb in Chinese and Korean traditional medicines. Considering the basicity of cyclopeptide alkaloids, a specific separation method was developed for an UHPLC system. The compounds were detected by DAD and ESI-qTOF-MS, and their fragmentation patterns were also acquired by MS^E technologies. Peak-picking of major compounds was performed with nine previously isolated compounds and two reference standard compounds. Tandem MS fragmentation behaviors of type-Ia and -Ib cyclopeptide alkaloids were investigated with the acquired data to develop a strategy for dereplication of other cyclopeptide alkaloid compounds in Z. jujuba var. spinosa. Two more cyclopeptide alkaloids were tentatively identified with UHPLC-ESI-qTOF-MS using this method.

• Bulletin of the Korean Chemical Society
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• v.32 no.4
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• pp.1183-1186
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• 2011

The unique Br signature was utilized for C-terminal amino acid sequencing of model peptides. C-terminal carboxyl group was selectively derivatized in peptides, containing side chain carboxyl group, using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) and Br was introduced using 4-bromophenylhydrazine hydrochloride (BPH) in a one pot reaction. Matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) tandem mass spectra were obtained carrying the Br signature in the y-series ions. The Br signature facilitated C-terminal sequencing and discrimination of C-terminal carboxyl groups in the free acid and amide forms.