• 분석과학
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• 제8권4호
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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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• 제26권2호
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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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• 제34권6호
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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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• 제15권5호
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• pp.537-540
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• 2000

일반적으로 paclitaxel과 유사한 구조를 가진 복잡한 유도체의 분리나 최종 제품에서의 엄격한 규정 등으로 인하여 한 단계의 HPLC system으로 정 제하는 것은 상당히 어렵다. 이러한 이유로 두 단계 HPLC system으로 식물세포 배양액으로 부터 paclitaxel의 대량 정제를 수행하였다. 즉, 첫번째 단계에 서 reverse-phase HPLC column, 두번째 단계에서 normal­p phase HPLC column을 사용하여 최종 제품을 얻었다. 또한 최종 정 제 된 paclitaxel 내 에 포함되어 있는 불순물 profile 확 언, 함량 분석, 그리고 이들 물질들에 대한 분리, 정제 및 동 정을 실시하였다 이들 불순물들 중에서 0.1% 이상되는 것은 모두 6종 (side chain derivative, baccatin III, 10-deacetyltaxol, cephalomannine, taxane derivative, 7-epitaxol)이며 NMR과 MS를 사용하여 화학구조를 분석한 결과 이미 알려진 pacli­t taxel 유도체 또는 전구체와 동일한 물칠로 밝혀 졌다. 본 연구 결과는 결국 paclitaxel 생산을 위한 품질관리 (quality con­t tr이)와 허가를 위한 자료로 유용하게 사용되어 진다.

• 한국식품과학회지
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• 제31권3호
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• pp.848-854
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• 1999

전통된장으로부터 안지오텐신전환효소(angiotensin converting enzyme; ACE)를 저해하는 물질을 추출하여 그 구조를 밝혀냈다. ACE 저해물질을 열수추출한 다음 gel permeation chromatography (GPC)를 통하여 ACE 저해작용이 큰 두 개의 큰 획분을 받았다. 앞획분은 90%와 70%의 ACE 저해효과를 나타내었으나 단일물질로 분리되지 않아 계속하여 역상 HPLC를 통하여 순수 분리를 하였다. 그러나 앞획분은 순수분리되지 않아 결국 2차원 전기영동/TLC를 통하여 분리한 결과 분자량이 759.63인 아미노기를 갖고 있는 비펩타이드 물질임이 밝혀졌다. 뒷획분은 다른 조건의 HPLC(reverse column과 $NH_2$, column)를 이용하여 순수분리에 성공하였다. 이중 ACE 저해효과가 큰 물질은 분자량 271.33인 dipeptide인 arginine-proline임을 밝혀냈다. 이물질의 ACE $IC_{50}$은 $92\;{\mu}M$이었다. 본 연구 결과는 대부분 ACE 저해물질이 3개 내지 7개 등의 긴 펩타이드임을 감안할 때, 짧은 dipeptide로 ACE 저해펩타이드가 한국의 전통된장에서 생산할 수 있음을 보여주고 있다.

• 한국환경농학회지
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• 제23권4호
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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.

• 한국식품조리과학회지
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• 제2권2호
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• pp.16-20
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• 1986

무우 속의 매운 맛 성분을 용매추출법 ($CCl_4$,사용)으로 분리하여 RP-HPLC법에 의해 정제한 후에 관능검사에 의해 pungency test를 하고, UV spectrum분석, GC/MC 분석을 수행한 결과 4-methylthio-3-butenyl isothiocyanate 임을 확인하였다. RP-HPLC에서 mobile Phase로 70% acetonitrile을 사용하였는데, 정제된 4-methylthio-3-butenyl isothiocfanate는 단일 peak로 분리되어 나왔고 이때의 retention time은 5.2분이였다. 아울러 RP-TLC에서 Rf치는 0.9이었다.

• KSBB Journal
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• 제7권1호
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• pp.21-26
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• 1992

미역엽상부의 MeOH추출물에 포함되어 있는 brassinosteroid활성물질을 검색하기 위하여 용매분획, CCD, silica gel흡착 chromatography, Sephadex LH-20 chromatography, charcoal 흡착 chromatography, Bondesil chromatography, 그리고 reverse phase의 HPLC등의 수법을 이용하여 분리 정제하고 각 단계에서 생물검정법으로 활성을 검정한 결과, 해조류인 미역에도 brassinosteroid활성물질의 존재가 인정되었는데, 그 함량은 타 식물의 영양조직에 비교하여 낮은 수준이었다.

• 한국응용과학기술학회지
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• 제15권4호
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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%).

• 약학회지
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• 제48권4호
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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.