• Proceedings of the Korean Society of Toxicology Conference
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• 2006.11a
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• pp.65-74
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• 2006

Modem drug discovery requires rapid pharmacokinetic evaluation of chemically diverse compounds for early candidate selection. This demands the development of analytical methods that offer high-throughput of samples. Naturally, liquid chromatography / tandem mass spectrometry (LC-MS/MS) is choice of the analytical method because of its superior sensitivity and selectivity. As a result of the short analysis time(typically 3-5min) by LC-MS/MS, sample preparation has become the rate- determining step in the whole analytical cycle. Consequently tremendous efforts are being made to speed up and automate this step. In a typical automated 96-well SPE(solid-phase extraction) procedure, plasma samples are transferred to the 96-well SPE plate, internal standard and aqueous buffer solutions are added and then vacuum is applied using the robotic liquid handling system. It takes only 20-90 min to process 96 samples by automated SPE and the analyst is physically occupied for only approximately 10 min. Recently, the ultra-high flow rate liquid chromatography (turbulent-flow chromatography)has sparked a huge interest for rapid and direct quantitation of drugs in plasma. There is no sample preparation except for sample aliquotting, internal standard addition and centrifugation. This type of analysis is achieved by using a small diameter column with a large particle size(30-5O ${\mu}$m) and a high flow rate, typically between 3-5 ml/min. Silica-based monolithic HPLC columns contain a novel chromatographic support in which the traditional particulate packing has been replaced with a single, continuous network (monolith) of pcrous silica. The main advantage of such a network is decreased backpressure due to macropores (2 ${\mu}$m) throughout the network. This allows high flow rates, and hence fast analyses that are unattainable with traditional particulate columns. The reduction of particle diameter in HPLC results in increased column efficiency. use of small particles (<2 urn), however, requires p.essu.es beyond the traditional 6,000 psi of conventional pumping devices. Instrumental development in recent years has resulted in pumping devices capable of handling the requirements of columns packed with small particles. The staggered parallel HPLC system consists of four fully independent binary HPLC pumps, a modified auto sampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The process of Bioanalytical Method Validation is required by the FDA to assess and verify the performance of a chronlatographic method prior to its application in sample analysis. The validation should address the selectivity, linearity, accuracy, precision and stability of the method. This presentation will provide all overview of the work required to accomplish a full validation and show how a chromatographic method is suitable for toxirokinetic sample analysis. A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method developed to quantitate drug levels in dog plasma will be used as an example of tile process.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.679-682
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• 2008

A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

• Korean Journal of Pharmacognosy
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• v.44 no.4
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• pp.350-361
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• 2013

Asarum sieboldii has been used for treatment of fever, pain, common cold, and chronic sinusitis in Korea. In this study, we performed quantification analysis of seven major constituents including aristolochic acid I, aristolochic acid II, ${\alpha}$-asarone, ${\beta}$-asarone, elemicin, methyl eugenol, and safrole in the 70% ethanol extract of Asarum sieboldii and its solvent fractions, n-hexane, ethylacetate, n-butanol, and water ones using a ultra-performance liquid chromatography-electrospray ionization-mass spectrometer(UPLC-ESI-MS) and gas chromatography-mass spectrometer(GC-MS). Regression equations of seven components were acquired with $r^2$ values >0.99. The values of limit of detection(LOD) and quantification(LOQ) were 0.1-3.9 ng/mL and 0.3-11.7 mg/mL, respectively. The amount of the seven compounds in Asarum sieboldii were not detected -143.66 mg/g. The established LC-MS/MS and GC-MS methods will be helpful to improve quality control of Asarum sieboldii.

• Food Science and Preservation
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• v.19 no.3
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• pp.438-442
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• 2012

To prepare calcium-binding peptides as calcium supplement, barley proteins were hydrolyzed using Flavourzyme for 18 h and the hydrolysates were ultra-filtered under 3 kDa as a molecular weight. The resultant filtered peptides were fractionated using ion exchange and normal-phase high performance liquid chromatography. Then each fraction that was obtained was determined for its calcium-binding activity to isolate the calcium-binding peptides. As a result, the highest calcium-binding peptide fraction was obtained, and the results suggest that barley protein hydrolysates can be used as a calcium supplement.

• Mass Spectrometry Letters
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• v.7 no.1
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• pp.12-15
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• 2016

Results of free and bound myo-inositol in infant formula (IF) are presented. Inositol was analyzed by HILIC ultra-performance liquid chromatography coupled with mass spectrometer. The levels of free myo-inositol in 27 Australian and 4 EU originated IF samples were 300-600 mg/kg of powder or 1.6-3.1 mg/100 kJ. The amount of bound inositol in lipid fraction of IF was, on average, 10% of free myo-inositol.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.05a
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• pp.5-5
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• 2010

Selection of specific medicinal sources as well as bioactive compounds is important for the preparation of medicine and related products with good quality. It is necessary to pay close attention for choosing correct medicinal sources, particularly in case of medicinal plants, because of their diversity, which can affect the quality and efficacy of medicine. Discrimination of plants based on morphological or genetic characteristics has been used as a conventional classification method of pharmaceutical sources so far; however, more need demands more general methods for accurate quality assessment of medicinal plants. In this study, ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) technique applied to this metabolic profiling is a powerful tool due to its higher sensitivity, resolution, and speed compared to conventional HPLC technique. The metabolite profiling of several medicinal plants including Panax ginseng was carried out using UPLC/Q-TOF MS and total metabolites were then subsequently applied to various statistical tools to compare the patterns. The developed metabolomics tool with UPLC/Q-TOF MS successfully identified and classified the samples tested according to their origins.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.245-251
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• 2021

Codonopsis lanceolata (Deoduk) was grown in East Asia, including Korea, China, Japan, and Russia, and the roots of C. lanceolata have been used as functional foods and traditional medicine to treat symptoms of cough, bronchitis, asthma, tuberculosis, and dyspepsia. The phytochemicals of C. lanceolata have been reported such as phenylpropanoids, polyacetylenes, saponins, and flavonoids that are involved in pharmacological effects such as anti-obesity, anti-inflammation, anti-tumor, anti-oxidant, and anti-microbial activities. Selecting marker substances of the main producing area by MS-based metabolomics analysis is important to ensure the beneficial effect of C. lanceolata without side-effects because differences in cultivated areas of plants were related not only to the safety of medicinal plants but also to changes in chemical composition and biological efficacy. In our present study, ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry combined with multivariate statistical analysis was applied to recognize the main producing area of C. lanceolata in South Korea. As a result of Principal Component Analysis and loading plot analysis of three groups, Inje (Kangwon-do), Hoengseong (Kangwon-do), and Muju (Jeonlabuk-do), several secondary metabolites of C. lanceolata including tangshenoside I, lancemaside A, and lancemaside G, were suggested as potential marker substances to distinguish the place of main producing area of C. lanceolata.

• Analytical Science and Technology
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• v.32 no.1
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• pp.24-34
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• 2019

The safety of food is occasionally questionable, as there have been some reports of products contaminated with illegal adulterants. In this study, the presence of 16 sedative-hypnotics and sleep inducers in dietary supplements was determined by ultra-high-performance liquid chromatography with UV detection (UPLC-UV) and quadruple Orbitrap mass spectrometry (Q-Orbitrap-MS). The UPLC method was validated, providing a linearity (R2) of more than 0.999, and LODs and LOQs that ranged from 0.2 to 0.5 and 0.6 to $1.5{\mu}g\;mL^{-1}$, respectively. The repeatabilities were 0.2-8.4 % (intra-day) and 0.3-4.5 % (inter-day), and the accuracies were 89.0-117.0 % (intra-day) and 87.8-111.9 % (inter-day). The mean recoveries of the spiked samples ranged from 98.7 to 107.3 %. The relative standard deviation (%RSD) of the stability was less than 2.4 %. Using the developed method, one sedative-hypnotic compound, phenobarbital, was detected in one of the nineteen samples tested. In addition, the major characteristic fragment ions of each target compound were confirmed using Q-Orbitrap-MS for higher accuracy. Monitoring the presence of these 16 sedative-hypnotics and sleep inducers in dietary supplements should be pursued in the interest of human health, and the results of this study confirmed that the developed method has value for this application.

• Journal of the Korean Society of Marine Environment & Safety
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• v.27 no.2
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• pp.340-354
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• 2021

The aim of this study was to investigate the spatial and temporal distribution of phytoplankton biomass and community composition in Jinhae Bay on the southern coast of Korea. Phytoplankton pigment analysis was conducted using ultra performance liquid chromatography (UPLC) were conducted from April to December 2019 at seven stations. Temperature, salinity, and dissolved oxygen (DO) and inorganic nutrients (dissolved nitrogen, dissolved phosphorus, and orthosilicic acid) were measured to investigate the environmental factors associated with the structure of phytoplankton community. Phytoplankton biomass (Chl-a) was the highest in July (mean 15.4±4.3 ㎍/L) and the lowest in December (mean 3.5±0.6 ㎍/L). Fucoxanthin was the most abundant carotenoid and showed a similar variation pattern to Chl-a, peridinin, and Chl-b. Phytoplankton community composition analysis showed that diatoms were a predominant group with an average abundance of 70 % whereas chlorophytes, cryptophytes, and dinoflagellates often appeared with lower averages. Further, the dominance of diatoms was closely correlated with water temperature and N:P ratio, which might be influenced by high temperatures in the summer and nutrient loading from the land. Additionally, freshwater and nutrient input by rainfall was estimated to be the most important environmental factor. Hence, the spatial and temporal variations in the composition of phytoplankton pigments and phytoplankton community were correlated with physicochemical and environmental parameters.

• Journal of Applied Biological Chemistry
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• v.65 no.4
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• pp.343-348
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• 2022

This study was performed to establish an analytical method for the standardization of Angelicae Dahuricae Radix as a functional ingredient. We established six compounds including oxypeucedanin hydrate (1), byakangelcol (2), oxypeucedanin (3), imperatorin (4), phellopterin (5) and isoimperatorin (6) as marker compounds of Angelicae Dahuricae Radix. An analytical method using Ultra Performance Liquid Chromatography (UPLC) was established and validated for marker compounds of Angelicae Dahuricae Radix. The specificity was confirmed by the chromatogram from UPLC and the value of coefficient determination was also higher than 0.999, indicating high linearity. The relative standard deviation (RSD) and recovery of marker compounds were less than 5% and in the range of 90- 110%, respectively, which means that this method has high accuracy and precision. Therefore, this analytical method could be used as basic data for the development of functional ingredients for health functional food of Angelicae Dahuricae Radix.