• Journal of Pharmaceutical Investigation
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• v.34 no.4
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• pp.327-331
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• 2004

A simple HPLC method with ultraviolet detection of ciprofloxacin in human plasma was developed and validated. After protein precipitation with trichloroacetic acid, chromatographic separation of ciprofloxacin in plasma was achieved at $50^{\circ}C$ with a $C_{18}$ column and methanol-phosphate mixture (pH 2.5), as mobile phase. Quantitative determination was performed by ultraviolet detection at 278 nm. The method was specific and validated with a limit of quantification of 100 ng/ml. The intra- and inter-day coefficients of variation were between 1.67% and 10.55% and accuracy between 92.01 % and 106.09%. The method has been successfully applied in a bioavailability study of 250 mg ciprofloxacin hydrochloride tablet.

• Analytical Science and Technology
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• v.19 no.3
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• pp.189-193
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• 2006

Chromatographic separation of glycerol monostearate, glycerol distearate and glycerol tristearate (GMS, GDS, and GTS) has been performed by normal phase HPLC method utilizing a Zorbax silica ($250{\times}4.6mm$, $5{\mu}m$) column and hexane-hexane, IPA and ethyl acetate mixtures as the eluent within 20 min. The observed reproducibility was less than 5% RSD, Suggesting that ELSD was an effective tool for detection of the glycerol stearates of low volatility without chromophore. The detection limits were in the concentration range of 0.3~2 mg/L, and the calibration curves (the log-log plots) were linear in the range of 4~1000 mg/L (with the slopes of 1.06~1.32). The application of the analytical procedure without pretreatment demonstrated that the proposed chromatographic method would be practical for a routine analysis of commercial products.

• Bulletin of the Korean Chemical Society
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• v.7 no.1
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• pp.45-50
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• 1986

Separation of optical isomers of some derivatives of amino acids by reversed-phase HPLC has been accomplished by adding a chelate of an optically active amino acid to copper(Ⅱ) to the mobile phase. Cu(Ⅱ) complexes of L-proline and L-hydroxyproline in the mobile phase showed different degrees of separation. Optical isomers of DNS derivatives of amino acids are selectively separated, but those of several other derivatives are not at all. The kinds of buffer agents, the pH, and the concentrations of acetonitrile and the Cu(Ⅱ) ligand all affect the separations. The elution behavior between D and L DNS-amino acids appears to depend on the alkyl side chain of the amino acids. A chromatographic mechanism is proposed that is based on a stereospecificity of the formation of ternary complexes by the D, L-DNS-amino acids and the chiral additive associated with the stationary phase. The steric effects of the ligand exchange reactions are related with the feasibility of cis and/or trans attack of the amino acids to the binary chiral chelate retained on the stationary phase.

• Bulletin of the Korean Chemical Society
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• v.13 no.3
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• pp.285-289
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• 1992

Enantiomeric separation of underivatized amino acids using N-alkyl-L-proline (octyl, dodecyl or hexadecyl) coated HPLC has been accomplished. The anchoring N-alkyl groups of L-proline provides a permanent adsorption of there solving chiral agent on the hydrophobic interface layer of a reversed phase. The factors controlling retention and enantioselectivity such as the Cu(II) concentration, pH of the eluent, the type and concentration of organic modifier in the hydroorganic eluent, and extent of coating were examined. The elution orders between D- and L-amino acids were consistent, L-forms eluting first, except histidine and asparagine. The extremely high enantioselectivity $(\alpha$ upto 13 for proline) is observed. The retention mechanism for the chiral separation can be illustrated by a complexation and hydrophobic interaction.

• Korean Chemical Engineering Research
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• v.46 no.4
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• pp.681-685
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• 2008

Chiral separation of racemic mandelic acid was achieved on a Kromasil KR100-5CHI-TBB column. Some chromatographic parameters (resolution, number of theoretical plates, capacity factor) are calculated under different separation conditions such as changes of mobile phase compositions (hexane/t-BME = 85/15 - 10/90) as well as formic acid concentrations for adjusting pH (0.1, 0.5, 1.0 v/v%). Flow rate versus number of theoretical plates was compared to evaluate column efficiency. To determine the adsorption isotherms, PIM (Pulse Input Method) was carried out. At the concentrations of racemic mandelic acid between 0.1 and 0.3 mg/ml, L- and D-mandelic acids have the same retention times of 8.8 and 9.4 min respectively. Mandelic acid isotherms show a linear form under the concentrations of 0.3 mg/ml with eluent (hexane/t-BME = 75/25). As the concentrations of mandelic acids increase, nonlinear Langmuir isotherms were observed as $C_{S,L}=3.358C_{M,L}/(1+0.0897C_{M,L})$ for L-mandelic acid and, $C_{S,D}=3.692C_{M,D}/(1+0.1457C_{M,D})$ for D-mandelic acid.

• Molecular & Cellular Toxicology
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• v.3 no.4
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• pp.273-281
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• 2007

Ceramide is involved in cell death as a lipid mediator of stress responses. In this study, we developed an improved method of ceramide quantification based on added synthetic ceramide and thin layer chromatography (TLC) separation, and applied to biological samples. Lipids were extracted from samples spiked with N-oleoyl-D-erythro-sphingosine ($C_{17}$ ceramide) as an internal standard. Ceramide was resolved by TLC, complexed with fatty-acidfree bovine serum albumin (BSA), and deacylated by ceramidase (CDase). The released sphingosine was derivatized with o-phthalaldehyde (OPA) and measured by high performance liquid chromatography (HPLC). The limit of detection for ceramide was about 1-2 pmol and the lower limit of quantification was 5 pmol. Ceramide recovery was approximately 86-93%. Ceramide concentrations were determined in biological samples including cultured cells, mouse tissues, and mouse and human plasma. TLC separation of ceramide provides HPLC chromatogram with a clean background without any interfering peaks and the enhanced solubility of ceramide by BSAceramide complex leads to the increased deacylation of ceramide. The use of an internal standard for the determination of ceramide concentration in these samples provides an accurate and reproducible analytical method, and this method can be applicable to diverse biological samples.

• Bulletin of the Korean Chemical Society
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• v.26 no.4
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• pp.529-536
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• 2005

Measurement of toxicologically relevant polychlorinated biphenyl (PCB) congeners such as non-ortho(IUPAC#) 77, 81, 126, 169 and mono-ortho 105, 114, 118, 123, 156, 157, 189 and di-ortho 170, 180 and polybrominated diphenyl ethers (PBDEs) such as 47, 66, 85, 99, 100, 138, 153, 154 in environmental samples become almost mandatory in several countries now. However, most of the available methods involve expensive instrumentations such as HRGC-HRMS or ECNI-LRMS, apart from expensive extraction and clean-up (with large volume of solvents) steps. A method has been devised combining the analytical separation power of PYE [2-(1-pyrenyl)ethyldimethysilylated silica] column HPLC and high-resolution gas chromatographic techniques including micro-electron capture detection (ECD) and two dimensional gas chromatograpy-ECD techniques to determine these eco-toxic substances at parts-per-trillion (ppt) levels. This combination resolves co-elution of congeners that occur in disproportionate ratios (e.g. CB-110 and -77) and allows accurate congener-specific determination of target compounds. This method is cost effective as it requires only hexane, that in small quantities (10 mL) and GC-ECD. The elution and analysis time are optimized to less man hours. This method is effectively utilized in the analysis of co-planar PCBs and PBDEs from archived solvent extracts of samples previously analyzed for pesticides and PCBs. Structure based separation of contaminant classes improves GCECD determination at ppt levels.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.5
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• pp.712-718
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• 2004

This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

• Natural Product Sciences
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• v.28 no.4
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• pp.168-180
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• 2022

Ephedra is a genus of the Ephedraceae family and is found in temperate regions, such as Central Asia and Europe. Among the various ephedra species, Ma Huang (Ephedra herb) is derived from the aerial parts of Ephedra sinica S tapf, Ephedra equisetina Bunge, and Ephedra intermedia Schrenk & C.A. Mey. Ma Huang contains various ephedra alkaloids, including (-)-ephedrine, (+)-pseudoephedrine, (-)-norephedrine, (+)-norpseudoephedrine, (-)-methylephedrine, and (+)-methylpseudoephedrine, which are found naturally as single enantiomers, although they can be prepared as racemates. Although the use of Ma Huang in foods is prohibited in Korea, products containing Ma Huang can be imported, and so it is necessary to develop a suitable analytical technique for the detection of Ma Huang in foods. Herein, we report the development of analytical methods for the detection of ephedra alkaloids in products containing Ma Huang. Following sample purification by solid phase extraction, quantitative analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). Additionally, the enantiomers were successfully separated using HPLC-DAD. We successfully analyzed various food samples, where the ephedra alkaloids were qualitatively and quantitatively determined, and the enantiomers were separated. It is expected that these methods may contribute toward preventing the distribution of illegal products containing Ma Huang.

• The Korean Journal of Food And Nutrition
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• v.14 no.6
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• pp.521-526
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• 2001

A method for the simultaneous and precise determination of lactone form and acid form monacolin K in red yeast rice by HPLC was developed in this study. The standard of acid form monacolin K was prepared by alkaline hydrolysis of its lactone form, which was purchased from Sigma company. The optimum HPLC system for the separation and quantification of acid form and lactone form monacolin K is based on the reversed-phase column, and the acidified mobile phase consisting of acetonitrile : 0.1% trifluoroacetic acid (TFA) water soln : 62 :38, the low limit detection amount was 5 ng (i.e.10 $\mu$l injection of 0.5 $\mu\textrm{g}$/ml) . And the optimal extracting system for monacolins in red rice was also presented here.