• Analytical Science and Technology
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• v.8 no.4
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• pp.859-864
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• 1995

Mono- and oligosaccharides are derivatized with p-aminobenzoic ethyl ester (ABEE), strongly absorbs UV light at 254 nm, in the presence of sodium cyanoborohydride. C18-bonded silica column is used for the separation of sugar-ABEE derivatives in an isocratic mode. RP-HPLC conditions are optimized by using ternary mixture as mobile phase and $45^{\circ}C$ as a column temperature. Sugar-ABEE derivatives are separated well within a short run time (ca. 25 min) by reverse-phase partition chromatographic mode. The ($1{\rightarrow}6$) linkage type of dihexose-ABEE derivatives has shorter retention time than ($1{\rightarrow}4$)-linkage type. After acid hydrolysis of glycoproteins with 2M trifluoroacetic acid, monosaccharide composition and contents are determined. This procedure is very useful for the simultaneous analysis of neutral and amino sugars in a single chromatographic step using RP-HPLC without reacetylation of deacetylated amino sugars, which are produced by acid hydrolysis.

• Journal of Pharmaceutical Investigation
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• v.26 no.2
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• pp.83-89
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• 1996

A reverse-phase HPLC method to determine DL-Carnitine Hydrochloride in pharmaceutical preparation is described. UV absorption derivatives of DL-Carnitine Hydrochloride were formed with p-Bromophenacyl Bromide in an essentially quantitative manner using crown ether as catalyst. The DL-Carnitine-bromophenacyl ester absorbed UV radiation strongly at 254nm, allowing the detection of as small a quantity as 12.5ng of DL-Carnitine Hydrochloride. A linear defection range was $5\;{\times}\;10-8 \;{\sim}\;5\;{\times}\;10-7M$ of DL-Carnitine Hydrochloride. And the linear regression at various drug concentration was =0.999 (n=10). The DL-Carnitine Hydrochloride in pharmaceutical preparation was successfully derivatized and separated from other constituents by reverse phase HPLC with detection at 254 nm.

• Bulletin of the Korean Chemical Society
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• v.34 no.6
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• pp.1745-1750
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• 2013

A novel green aqueous mobile phase modified with room temperature ionic liquids (RTILs) was employed in the absence of volatile organic solvents or ion-pairing reagents to analyze thiamine, a very polar compound, by reverse phase high performance liquid chromatography (RP-HPLC). Due to its strongly hydrophilic nature, thiamine was eluted near the column dead time ($t_0$) using a mobile phase without adding RTILs or ion-pairing reagents, even if a 100% aqueous mobile phase, which has weak elution power under reverse phase conditions, was used. Thus, 1-ethyl-3-methyl-imidazolium hexafluorophosphate ([EMIM][$PF_6$]), which has the strongest chaotropic effect, was selected as a mobile phase additive to improve retention and avoid baseline disturbances at $t_0$. Various mobile phase parameters such as cation moiety, chaotropic anion moiety, pH and concentration of RTILs were optimized to determine thiamine at the proper retention time. Method validation was performed to assess linearity, intra- and inter-day accuracy and precision, recovery and repeatability; all results were found to be satisfactory. The developed method was also compared to the current official United States Pharmacopoeia (USP) and Korean Pharmacopoeia (KP) methods using an organic mobile phase containing an ionpairing reagent by means of evaluating various chromatographic parameters such as the capacity factor, theoretical plate number, peak asymmetry and tailing factor. The results indicated that the proposed method exhibited better efficiency of thiamine analysis than the official methods, and it was successfully applied to quantify thiamine in pharmaceutical preparations.

• KSBB Journal
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• v.15 no.5
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• pp.537-540
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• 2000

In developing a HPLC purification process, it was hoped that a single chromatographic system would be sufficient to abtain pure paclitaxel in high yield. However, no such system was found, due in part to the complex taxoid profile of crude paclitaxel and to the rigorous nature of the product specification. A two step HPLC purification was adopted using reverse-phase separation on C(sub)18 as a first step, and normal-phase separation on silica as the final polishing step. Impurity profiles were established and maintained for paclitaxel, which identified and quantified each impurity observed in purified paclitaxel from these two steps, all impurities at or above 0.1% were identified. Results provide information for improving the quality control of paclitaxel production.

• Korean Journal of Food Science and Technology
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• v.31 no.3
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• pp.848-854
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• 1999

Inhibitory compounds of angiotensin converting enzyme (ACE) were separated from Doenjang (traditional Korean fermented soybean paste). Water extracts from Doenjang which showed ACE inhibitory activity were separated with gel permeation chromatography (GPC), in which two fractions with high ACE inhibitory activities were obtained. The first fraction from GPC was further isolated by semi-preparative reverse phase preparative-HPLC (high performance liquid chromatography) and 2-dimensional electrophoresis/thin layer chromatography (TLC). The purified spot had molecular weight of 759 daltons and ninhydrin-positive non-peptide. The second fraction from GPC was also further isolated by semi-preparative reverse phase HPLC and $NH_2-column$ HPLC. One fraction with high ACE inhibitory activity was purified and characterized. Molecular weight of this fraction by LC-MS was 272.34 daltons. The active fraction was identified as Arg-Pro with ACE $IC_{50}$ of $92\;{\mu}M$.

• Korean Journal of Environmental Agriculture
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• v.23 no.4
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• pp.245-250
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• 2004

An analytical method was developed to determine monocrotophos residues in apple, citrus, and soil using high-performance liquid chromatography (HPLC) with ultraviolet absorption detection. Monocrotophos was extracted with acetone from apple, citrus and moist soil samples. The extract was concentrated, added with saline water, and subjected to n-hexane washing to remove nonpolar co-extractives. Dichloromethane partition was then followed to recover monocrotophos from the aqueous phase. Silica gel column chromatography was employed to further purify the extract prior to HPLC determination. Reverse-phase HPLC using an oct-adecylsilyl column was successfully applied to separate and quantitate the monocrotophos residue in sample extracts at the wavelength of 230 nm. Overall recoveries of monocrotophos from fortified samples averaged $95.3{\pm}2.1%$ (n=6), $970{\pm}0.7%$ (n=6), and $92.8{\pm}4.3%$ (n=12) for apple, citrus, and soil, respectively. The proposed method was quite reproducible and sensitive enough to replace the troublesome gas-liquid chromatographic analysis for monocrotophos residues.

• Korean journal of food and cookery science
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• v.2 no.2
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• pp.16-20
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• 1986

The major pungent component from Korean radish roots was purified by reverse phase high performance liquid chromatography (RPHPLC), and characterized as 4-methylthio-3-butenyl isothiocyanate on the basis of the sensory test (pungency), UV spectrum and mass spectrum analysis. The purified isothiocyanate moved as a single peak(retention time, 5.2 min) in RP-HPLC analysis, and as a single spot(Rf, 0.9) in TLC analysis.

• KSBB Journal
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• v.7 no.1
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• pp.21-26
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• 1992

In order to examine the presence of brassinosteroid substance in sea mustard(Undaria pinnatifida), leaves of sea mustard were extracted with MeOH. The extract was purified by slovent fractionation, counter current distribution, silica gel adsorption chromatography, charcoal adsorption chromatography, Bondesil chromatography, and reverse phase HPLC, successively. The activity was monitored by the rice inclination test and its prescence could be confirmed in each purification step. Although sea mustard contained a less amount of the active substance than the vegetative tissue of higher plants, brassinosteroid was clearly present endogenously in sea mustard. We acknowledge that our work is probably the first publication reporting the presence of brassinosteroid in marine algae plants.

• Journal of the Korean Applied Science and Technology
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• v.15 no.4
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• pp.1-9
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• 1998

The lipids from the seeds of Pinus koraiensis mostly composed of triacylglycerols (TGs), in which linoleic acid (46.2 mol%) and oleic acid (25.6 mol%) are present as main components in the fatty acid composition. Surprisingly, they also have unusual fatty acids with ${\Delta}^5$-double bond systems such as ${\Delta}^{5.9.12}-C_{18:3}$ (16.0 mol%), ${\Delta}^{5.9}-C_{18:2}$ (2.3 mol%) and ${\Delta}^{5.11.14}-C_{20:3}$ (0.8 mol%). Saturated fatty acids of palmitic, stearic and arachidic acid were present in less than 8.0 mol%. TG was resolved into 17 fractions by reverse-phase HPLC according to so-called partition number (PN) suggested by Plattner, in which TG molecules with ${\Delta}^{5}$-NMDB acyl chains eluted later than did those with ${\Delta}^{9}$-MDB acyl radicals. $Ag^+$-HPLC separated the TG into 14 fractions more clearly than did those with ${\Delta}^{9}$-MDB acyl radicals. $Ag^+$-HPLC separated the TG into 14 fractions more clearly than did reverse-phase HPLC, and the complexity of ${\Delta}^{5.9.12}-C_{18:3}$ moiety with silver ion impregnated in the column bed was in the level between ${\Delta}^{9.12.15}-C_{18:3}$ ($C_{18:3{\omega}3}$) and $C_{18:2{\omega}6}$ (${\Delta}^{9.12}-C_{18:2}$). In the $Ag^+$-HPLC, it was found that the molecular species having a given-numbered double bonds widely spreaded in the acyl chains eluted earlier than those concentrated in one acyl chain. The main molecular species are $(C_{18:2{\omega}6})_2/{\Delta}^{5.9.12}-C_{18:3}$ (14.8 mol%), $C_{18:1{\omega}9}/C_{18:2{\omega}6})_2$ (12.8 mol%) and $C_{18:1{\omega}9}/C_{18:2{\omega}6}/{\Delta}^{5.9.12}-C_{18:3}$ (10.9 mol%).

• YAKHAK HOEJI
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• v.48 no.4
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• pp.231-235
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• 2004

A new high performance liquid chromatographic method was applied for the determination of hyperin in Acanthopanax senticosus and A sessiliflorus. The stationary phase used was $\beta$-Bondapak $C_{18}$ reverse-phase column and a mobile phase program was a gradient of acetonitrile and distilled water at a flow rate of 1.0 ml/min. Hyperin was detected at 210 nm, and the analysis was successfully carried out within 20 min. Hyperin was detected in the one year-grown and two years-grown stem of A senticosus (0.47 and 0.13 mg/g, respectively) and A sessiliflorus (0.14 and 0.03 mg/g, respectively). Hyperin was detected in the main and branch root of A sessiliflorus (0.30 and 0.09 mg/g, respectively). But there is no detection of hyperin in the main and branch root of A senticosus.