• 한국식품영양과학회지
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• 제22권6호
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• pp.815-822
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• 1993

This experiment was carried out to determine the separation condition of 1-dimethylaminonaphthalene-5-sulfony(Dansyl) derivatives of amino acids by reverse-phase high performance liquid chromatography with Nova-Pak C18 column. Determined solvent system was solvent A(200mA phosphate buffer pH 6.8 15%, acetonitrile 11%, water 74%) and solvent B(acetonitrile 65%, methanol 28%, water 7%). Linear gradient of solvent B was applied from 12% to 80% for 50min. Complete separation of 20 amino acids including asparagine and glutamine which constitute protein was achieved within 50min. As the detection limit was the range of picomole, the resolution power was excellent. Reproducibility of the retention time was less than mean $\pm$0.05min. According to the above optimum chromatographic conditions, the amino acid composition of some food and human blood was examined. The most affluent amino acid was alanine in human blood, aspartic acid and glutamic acid in soy sauce, alanine and threonine in soy milk and proline in milk and yoghurt.

• Mass Spectrometry Letters
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• 제12권1호
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• pp.1-10
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• 2021

A simple, specific, and economical LC-MS/MS method was investigated for the screening of 43 prescribed antihypertensive and related drugs in human urine. The urine samples were simply prepared by diluting and mixing with internal standard before directly introduced to the LC-MS/MS system, which is fast, straightforward, and cost-effective. Fractional factorial, Box-Behnken, and I-optimal design were applied to screen and optimize the mass spectrometric and chromatographic factors. The analysis was carried out on a triple quadrupole mass spectrometer system utilizing multiple reaction monitoring with positive and negative electrospray ionization method. Chromatographic separation was performed on a Thermo Scientific Accucore RP-MS column (50 × 3.0 mm ID., 2.6 ㎛) using two separate gradient elution programs established with the same mobile phases. Chromatographic separation was performed within 12 min. The optimal method was validated based on FDA guideline. The results indicated that the assay was specific, reproducible, and sensitive with the limit of detection from 0.1 to 50.0 ㎍/L. The method was linear for all analytes with coefficient of determination ranging from 0.9870 to 0.9981. The intra-assay precision was from 1.44 to 19.87% and the inter-assay precision was between 2.69 and 18.54% with the recovery rate ranges from 84.54 to 119.78% for all drugs measured. All analytes in urine samples were stable for 24 h at 25℃, and for 2 weeks at -60℃. The developed method improves on currently existing methods by including larger number of cardiovascular medications and better sensitivity of 12 analytes.

• Journal of Ginseng Research
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• 제44권6호
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• pp.784-789
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• 2020

Background: The separation of isomeric compounds from a mixture is a recurring problem in chemistry and phytochemistry research. The purification of pharmacologically active ginsenoside Rb₃ from ginseng extracts is limited by the co-existence of its isomer Rb₂. The aim of the present study was to develop an enzymatic elimination-combined purification method to obtain pure Rb₃ from a mixture of isomers. Methods: To isolate Rb₃ from the isomeric mixture, a simple enzymatic selective elimination method was used. A ginsenoside-transforming glycoside hydrolase (Bgp2) was employed to selectively hydrolyze Rb₂ into ginsenoside Rd. Ginsenoside Rb₃ was then efficiently separated from the mixture using a traditional chromatographic method. Results: Chromatographic purification of Rb₃ was achieved using this novel enzymatic elimination-combined method, with 58.6-times higher yield and 13.1% less time than those of the traditional chromatographic method, with a lower minimum column length for purification. The novelty of this study was the use of a recombinant glycosidase for the selective elimination of the isomer. The isolated ginsenoside Rb₃ can be used in further pharmaceutical studies. Conclusions: Herein, we demonstrated a novel enzymatic elimination-combined purification method for the chromatographic purification of ginsenoside Rb₃. This method can also be applied to purify other isomeric glycoconjugates in mixtures.

• Bulletin of the Korean Chemical Society
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• 제35권2호
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• pp.539-545
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• 2014

Styrene-acrylamide co-polymer was immobilized on porous partially sub-$2{\mu}m$ silica monolith particles and inner surface of fused silica capillary ($50{\mu}m$ ID and 28 cm length) to result in ${\mu}LC$ and CEC stationary phases, respectively, for separation of anomeric D-glucose derivatives. Reversed addition-fragmentation transfer (RAFT) polymerization was incorporated to induce surface polymerization. Acrylamide was employed to incorporate amide-functionality in the stationary phase. The resultant ${\mu}LC$ and CEC stationary phases were able to separate isomers of D-glucose derivatives with high selectivity and efficiency. The mobile phase of 75/25 (v/v) acetonitrile (ACN)/water with 0.1% TFA, was used for HPLC with a packed column (1 mm ID, 300 mm length). The effects of pH and ACN composition on anomeric separation of D-glucose in CEC have been examined. A mobile phase of 85/15 (v/v) ACN/30 mM sodium acetate pH 6.7 was found the optimized mobile phase for CEC. The CEC stationary phase also gave good separation of other saccharides such as maltotriose and Dextran 1500 (MW~1500) with good separation efficiency (number of theoretical plates ~300,000/m).

• Journal of Nutrition and Health
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• 제5권4호
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• pp.145-150
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• 1972

In order to establish a new biochemical parameter to evaluate nuitrtitional status. the present paper presents a paper chromatographic procedure. which enables clear separation of the essential and nonessential free amino acids in the urine without desalting the sample prior to chromatographic analysis The children of school age. showing difference in their body physiques. from the rural and urban areas in Korea were selected in collecting the 24 hour urine specimens. with the following conclusions. (1) A new procedure utilyzing paper chromatographic analysis. without desalting the urine. was proposed to establish a new biochemical parameter for nutritional status by analyzing the ratio of the nonessential to the essential free amino acids in urine. (2) According to the pesent procedure. the ratio obtained with the urban children was proved to be $1.33{\pm}0.18$, whereas that with the rural children was $1.59{\pm}0.24$; suggesting the lower the ratio. the superior the body physiques and nutritional status. (3) The ratio had a statistically significant correlation with the body and sitting heights as well as the body weight. but not with the girth.

• Bulletin of the Korean Chemical Society
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• 제27권2호
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• pp.291-294
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• 2006

A highly sensitive high-performance liquid chromatographic-tandem mass spectrometric method (HPLC-MSMS) has been developed to quantify itraconazole in human plasma for the purpose of pharmacokinetic studies. Sample preparation was carried out by liquid-liquid extraction using loratadine as an internal standard. Chromatographic separation used a YMC $C_{18}$ column, giving an extremely fast total run time of 3 min. The method was validated and used for the bioequivalence study of itraconazole tablets in healthy male volunteers (n = 31). The lower limit of detection proved to be 0.2 ng /mL for itraconazole.

• Bulletin of the Korean Chemical Society
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• 제31권3호
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• pp.620-623
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• 2010

A brush type chiral stationary phase (CSP 2) derived from ${\alpha}$-amino ${\beta}$-lactam was prepared for the separation of the enantiomers of ${\beta}$-blockers. Compared to the CSP derived from ${\alpha}$-amino phosphonate (CSP 1), in general, the conformationally rigid CSP 2 showed greater scope and much enhanced enantioselectivity for the resolution of ${\beta}$-blockers. The effect of various salt additives on enantioseparation of ${\beta}$-blockers in the mobile phase was investigated. The unusual effect of temperature on the chromatographic behaviors was observed on CSP 2. It also afforded appreciable increases in enantioselectivity without significantly affecting resolution, as the column temperature was reduced.

• 대한화학회지
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• 제8권2호
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• pp.62-64
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• 1964

The cation exchange chromatographic studies for the analysis of transition metals have been described. The quantitative separation of a mixture of Fe(Ⅲ), Cu(II), Zn(II), Ni(II), Cd(II), Co(II) and Mn(II) has been obtained by elution, through a 28cm column of the resin, Dowex 50 ${\times}$ 4 (100∼200 mesh), using 0.45 M $NaNO_3$+0.05 M Na-tartrate solution as eluent, starting with the eluent of pH 3.5, followed stepwise by pH 4.0 and 4.5. A comparison between the calculated and the observed peak positions in the elution curve has been shown. The relative stability constants for tartrate complexes of some transition metals have been calculated by using distribution ratios obtained in this separation procedure.

• Bulletin of the Korean Chemical Society
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• 제25권9호
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• pp.1336-1340
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• 2004

The same kind of chiral stationary phase with a commercialized chiral column was used to make preparative chiral columns and was applied to resolve racemic N-acetyl-1-naphthylethylamide (3) by preparative liquid chromatography. An improved chromatographic condition to resolve racemic 3 on the CSP was examined by changing flow rate and kind of the mobile phase and the sample injection volume. The optimized separation conditions were applied to resolve racemic 1,1'-Binaphthyl-2,2'-diamine(4).

• 약학회지
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• 제30권1호
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• pp.47-50
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• 1986

The enantiomers of five amino acids (alanine, valine, threonine, leucine and phenylalanine) could be separated by gas chromatography with optically active (S)-5-isopropyll-$N^3$-phenyl-2-thiohydantoinic stationary phase, which prepared from L-valine and phenylisothiocyanate. Gas chromatographic separations on methylesterificated and N-trifluoroacetylated amino acids have been conducted in isothermal at several column temperatures (180~190, 200, $210^{\circ}C$). The separation factors were 1.29 (alanine, $190^{\circ}C$), 1.35 (valine, $190^{\circ}C$), 1.33 (threonine, $190^{\circ}C$), 1.17 (leucine, $190^{\circ}C$) and 1.05 (phenylalanine, $190^{\circ}C$) and D-isomers eluted prior to L-isomers in every instance. The result of this experiment shows that this stationary phase can be used for the separation of the other amino acids enantiomers.