• Analytical Science and Technology
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• v.24 no.3
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• pp.173-182
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• 2011

This research examined prostanozol and its metabolites in urine of women who took the medicine (prostanozol). Prostanozol and its metabolites were successfully separated and detected by using LC/ESI/MS and GC/TOF-MS. Mass spectrum of LC/ESI/MS estimated molecular weight of Prostanozol and its metabolites and that of GC/TOF-MS verified them. For M1, carbon number 17 of Prostanozol substituted to a keto group and it is called 17-keto-Prostanozol. M2 turned out to be hydroxy-17-keto-Prostanozol. It came from substitution of one hydroxyl group of pyrazole nucleus and A-ring of M1. Substitution of one hydroxyl group of B-ring or C-ring became M3, hydroxy-17-keto-Prostanozol. M4 was found to be a hydroxy-17-keto-Prostsnozol transposed from one hydroxyl group to a D-ring. M5 has a hydroxyl group of carbon number 17. One hydroxyl group is substituted from B-ring or C-ring and it is assumed to be hydroxy-17-hydroxy-Prostanozol. M6 was turned out to be dihydroxy-17-keto-Prostanozol transposed from one hydroxyl group to pyrazole nucleus or A-ring and to B-ring or C-ring. Like M6, M7 has a keto group at carbon number 17 and was identified as dihydroxy-17-keto-Prostanozol. M7 has one hydroxyl group at pyrazole nucleus or A-ring and also at D-ring. At last M8 was found to be dihydroxy-17-hydroxy-Prostanozol. Pyrazole nucleus or A-ring has got one hydroxyl group and other rings were substituted to another hydroxyl group. From above, M5, M7 and M8 were verified as new metabolites that were not discovered yet. Prostanozol and all of the 8 metabolites formed glucuronic conjugates as a result of conjugation reaction test in human body. Some of 8 metabolites were excreted without forming conjugates. Particularly M6 and M7 were excreted as sulfate conjugates.

• Mass Spectrometry Letters
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• v.8 no.4
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• pp.114-118
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• 2017

While saliva can be considered as good biological fluid for monitoring biomarkers due to many advantages including its communication with blood and the non-invasive nature during its sampling, its applications to that purpose is still limited. As a part of efforts to expand the applications of saliva to the protein biomarker research, we carried out global absolute quantitation of proteins in human whole saliva (WS) by bottom-up proteomics techniques mainly based on nLC-Q-IMS-TOF employing $MS^E$. From the analyses of a pooled WS sample collected from 22 healthy Korean volunteers, 93 proteins ranging from $5.89{\times}10^1ng/mL$ (immunoglobulin heavy chain) to $1.59{\times}10^4ng/mL$ (${\alpha}-amylase$ 1) were confirmed. For the validation of the present results, human serum albumin in the same sample was quantitated by ELISA and its result was compared with that from the nLC-Q-IMS-TOF study. As a result, there was no significant difference between two results from individual approaches ($1.18{\times}10^4{\pm}0.03{\times}10^4 ng/mL$ from nLC-Q-IMS-TOF experiments vs. $1.23{\times}10^4{\pm}0.07{\times}10^4ng/mL$ from ELISA experiments, n=3, p=0.309). To our knowledge, this is the first global absolute quantitation of proteins in human whole saliva and information from the present study can be widely used as the first level reference for the discovery of new protein biomarkers from human whole saliva as well as for quantitative applications of human whole saliva proteins.

• YAKHAK HOEJI
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• v.57 no.4
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• pp.250-257
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• 2013

Biosimilars are important drugs in medicine and contain many glycosylated proteins. Thorough analysis of the glycosylated protein is a prerequisite for evaluation of biosimilar glycan drugs. A method to assess the diversity of N-glycosylation sites and N-glycans from biosimilar glycan drugs has been developed using two separate methods, LC-MS/MS and MALDI-TOF MS, respectively. Development of sensitive, accurate, and efficient methods for evaluation of glycoproteins is still needed. In this study, analysis of both N-glycosylation sites and N-glycans of glycoprotein was performed using the same LC-MS/MS with two different nano-LC columns, nano-C18 and nano-porous graphitized carbon (nano-PGC) columns. N-glycosylated proteins, including RNAse B (one N-glycosylation site), Fetuin (three sites), and ${\alpha}$-1 acid glycoprotein (four sites), were used, and small amounts of each protein were used for identification of N-glycosylation sites. In addition, high mannose N-glycans (one type of typical glycan structure), Mannose 5 and 9, eluted from RNAse B, were successfully identified using nano-PGC-LC MS/MS analysis, and the abundance of each glycan from the glycoprotein was calculated. This study demonstrated an accurate and efficient method for determination of N-glycosylation sites and N-glycans of glycoproteins based on high sensitive LC-MS/MS using two different nano-columns; this method could be applied for evaluation of the quality of various biosimilar drugs containing N-glycosylation groups.

• Journal of the Korean Magnetic Resonance Society
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• v.22 no.2
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• pp.26-33
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• 2018

Xanthium sibiricum is used as a traditional folk medicine for the treatment of cancer, fever, headache, nasal sinusitis, and skin pruritus. This study aimed to identify components from Xanthium sibiricum extracts using an SPE-800MHz NMR-MS hyphenated system. The simultaneous acquisition of MS and NMR spectra from the same chromatographic peaks significantly increases the depth of information acquired for the compound and allows the elucidation of structures that would not be possible using MS or NMR data alone. LC -NMR analysis was conducted using a HPLC separation system coupled to 800 MHz spectrometer equipped with a cryoprobe, and a SPE unit was used to automatically trap chromatographic peaks using a HPLC pump. LC-MS analysis was conducted with a Q-TOF MS instrument using ESI ionization in the negative ion mode. Using the hyphenated analysis, several secondary metabolites were identified, such as 3',5'-O-dicaffeoylquinic acid, 1',5'-O-dicaffeoyl- quinic acid, and ethyl caffeate. These results demonstrate that the SPE-800MHz NMR-MS hyphenated system can be used to identify metabolites within natural products that have complex mixtures.

• Reproductive and Developmental Biology
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• v.37 no.4
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• pp.247-253
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• 2013

Here, we present an approach of blood plasma proteome profiling and their comparisons between the young and the adult pigs as prerequisite for the identification of bio-markers related to the health conditions, growth performance and meat quality. To profile the proteome in porcine plasma, blood samples were collected from 19 young piglets and 20 adult male barrows and the plasma was retrieved. Then, protein profiling was initiated using one and two-dimensional electrophoresis. Proteins were spotted and then identified by MALDI-TOF-TOF and LC-MS-MS. In the results, more than thirty-six and twenty eight protein spots were selected in young piglets and adult pigs, respectively and twenty three proteins were identified. The proteome profile images were compared between those ones using Image Master Version 7.0. The image of expressed proteome showed that most of proteins from plasma of young piglet separated clearly and concentrated in 2DE display compared to ones from adult. Image analysis in detail was carried out to look for the specific proteins related to age progression. It demonstrated that the characteristics of proteome expression could be distinct to their age stages. Further investigations needed to proceed to understand the age dependent change of protein conformation and biological meaning of those differences in proteome expression between young and mature adult pigs.

• Biomedical Science Letters
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• v.23 no.4
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• pp.395-401
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• 2017

Various preclinical and clinical trials have been conducted the efficacy of celastrol. In data presented in the current manuscript is the first trial to inhibit Helicobacter pylori with celastrol. In this study, the quantitative change of various H. pylori proteins including CagA and VacA by the anti-bacterial effect of celastrol was determined. The anti-H. pylori effects of celastrol was investigated by performing 2-dimensional electrophoresis and additional supporting experiments. After 2-dimensional electrophoresis analysis, spot intensities were analyzed and then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). The results show that celastrol has multiple effects on protein expression in H. pylori.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.4
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• pp.286-291
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• 2015

Equol has beneficial effects on human health. Fermented soy products contain equol, and many microbes participate in the equol production process. This study investigated fermented Korean soybean paste, doenjang. Thirty seven doenjang samples collected from different manufacturers were examined. Equol was detected in 3 samples (D2, D13, and D19) at the maximum content of 507 ng/100 g by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fifteen microbial species were isolated and identified by 16S rRNA gene sequence analysis and by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Bacillus spp, Paenibacillus spp, Tetragenococcus spp, Stapylococcus spp, and Clostridium species were the predominant bacteria in equol containing doenjang samples.

• Journal of Sensor Science and Technology
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• v.21 no.1
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• pp.13-20
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• 2012

Due to its high conductivity, pyrylium has been frequently used in electron transfer reactions or in the synthesis of various organic materials. It has also been used as a sensor material. Traditionally, the compounds have been synthesized using various methods; mostly in a multiple steps. In this study, two pyrylium salts, 2, 6-di-(4'-methylphenyl) pyrylium fluoroborate and perchlorate were synthesized. The synthesis of these products was confirmed by 1H-NMR, LC/TOF-MS and FT-IR analyses while their photo-properties were analyzed using UV/VIS spectrophotometry. In addition, the electron transfer capacities of the salts were analyzed with a conductivity meter, it was found that their electron conductivities were high. When the synthesized compounds were dissolved in acetone, a green fluorescent material was observed to form. The fluorescent material can be used as a sensitizer in the electrical industry.

• Journal of Food Hygiene and Safety
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• v.33 no.5
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• pp.339-346
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• 2018

This study was conducted to monitor the current status of pesticide residues in a total of 297 agricultural products for raw juice from January 2017 to September 2017. Pesticide residues in samples were analyzed by multiresidue method for the determination of multiclass pesticide with an aim to detect 220 pesticides using GC/ECD, GC/NPD, TOF/MS, LC/PDA, LC/FLD, and LC/MS-MS. The analysis revealed the presence of nine pesticides in 14 samples. The detected pesticides were chlorpyrifos, cyflufenamid, chlorothalonil, pendimethalin, azoxystrobin, pyridalyl, imazalil, fludioxonil and procymidone. The kinds of detected crops were mandarin, carrot, apple, celery, chicory, orange and grape. Risk assessment evaluated human health exposure with the ratio of EDI (Estimated daily intake) to ADI (Acceptable daily intake) of pesticides detected and the range of %ADI (the ratio of EDI to ADI) values was 0.0011~6.7087% and safety level.

• Bulletin of the Korean Chemical Society
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• v.34 no.9
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• pp.2663-2670
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• 2013

The purpose of stability testing is to provide evidence about how the quality of a drug varies with time under the influence of a variety of environmental factors. In this study, erythropoietin (EPO) was analyzed under different pH (pH 3 and pH 9) and temperature ($25^{\circ}C$ and $40^{\circ}C$) conditions according to current Good Manufacturing Practice (cGMP) and International Conference on Harmonisation (ICH) guidelines. The molecular weight difference between intact EPO and deglycosylated EPO was determined by SDS-PAGE, and aggregated forms of EPO under thermal stress and high-pH conditions were investigated by size exclusion chromatography. High pH and high temperature induced increases in dimer and high molecular weight aggregate forms of EPO. UPLC-ESI-TOF-MS was applied to analyze the changed modification sites on EPO. Further, normal-phase high-performance liquid chromatography was performed to identify proposed glycan structures and high pH anion exchange chromatography was carried out to investigate any change in carbohydrate composition. The results demonstrated that there were no changes in modification sites or the glycan structure under severe conditions; however, the number of dimers and aggregates increased at $40^{\circ}C$ and pH 9, respectively.