• Journal of Applied Biological Chemistry
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• 제55권1호
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• pp.61-65
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• 2012

Three major curcuminoids in turmeric (Curcuma longa), curcumin, demethoxycurcumin, and bisdemethoxycurcumin, were efficiently extracted by optimizing extraction condition and simultaneously analyzed and identified using reverse phase high-performance liquid chromatography, thin-layer chromatography, and liquid chromatography-mass spectrometry method. The highest yield of extraction amount 0.279 g, 9.30% was obtained by dipping method with extraction time of 7 h.

• Natural Product Sciences
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• 제24권4호
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• pp.288-292
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• 2018

High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase highperformance liquid chromatography (RP-HPLC) method was developed to isolate dihydrophaseic acid 3'-O-${\beta}$-D-glucopyranoside (DHPAG) from the extract of Nelumbo nucifera seeds. Enriched DHPAG sample (2.3 g) was separated by HPCCC using ethyl acetate/n-butanol/water system (6:4:10, v/v/v, normal-phase mode, flow rate: 4.0 mL/min) to give 23.1 mg of DHPAG with purity of 88.7%. Further preparative RP-HPLC experiment gave pure DHPAG (16.3 mg, purity > 98%). The current study demonstrates that utilization of CCC method maximizes the isolation efficiency compared with that of solid-based conventional column chromatography.

• 생약학회지
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• 제24권4호
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• pp.309-312
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• 1993

A new method for separation and quantitative determination of isoflavone glucosides in rhizomes of Belamcanda chinensis (Iridaceae) by high performance liquid chromatography was elaborated. A reverse-phase system with a Spheri-5 RP-18 column using MeOH:HOAc:$H_2O$(24 : 5 : 71) as a mobile phase was developed. The isoflavonoids were detected at 268 nm and the analysis was successfully carried out within 15 min.

• 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.

• 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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• 제25권3호
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• pp.205-214
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• 2010

역류크로마토그래피 (counter-current chromatography, CCC)는 일련의 분배과정을 한 개의 튜브 내에서 연속적으로 일어나도록 고안된 시스템으로서 컬럼으로는 polytetrafluoroethylene(PTFE) 튜브가 다층으로 감겨있는 원통형의 홀더 3개가 서로 기어를 통해 물려있으며, 홀더가 회전과 공전을 통해 튜브의 꼬임을 방지하는 rotary seal-free flow centrifuge 시스템으로 되어있다. 역상 HPLC (reverse phase HPLC)에서는 고정상이 실리카에 결합된 유기물단 (organic moiety)이 수용성 이동상 물질에 의해서 용매화 (solvated)되는 반면 CCC는 실리카 대신에 강한 중력장에 의해 분리되는 자유로운 용매가 고정상이 되며 이 고정상의 부피비율은 20-30%에 이른다. 즉 고체담체에 결합된 유기관능기 대신에 물과 섞이지 않는 hexane 같은 유기용제가 고정상으로 사용되는 것이다. 고속역류크로마토그래피 (high-performance countercurrent chromatography, HPCCC)는 CCC의 기능을 향상시킨 분리시스템으로서 높은 중력장하에서 높은 이동상 속도와 높은 분리효율과 짧은 분리시간을 특징으로 하고 있다. 특히 mg 단위에서 kg 단위로의 스케일업이 선형적으로 가능하다는 큰 장점을 지니고 있다. 이 총설에서는 현재까지 개발된 CCC의 일반적인 이론을 간략히 정리하고 최신 HPCCC 장비의 적용 예를 살펴보고 그 응용분야로서 생리활성물질의 분리 및 정제와 관련된 연구동향을 정리하였다.

• 한국약용작물학회지
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• 제9권1호
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• pp.21-25
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• 2001

인삼의 주종 사포닌인 7종 사포닌($Rb_1,\;Rb_2,\;Rc,\;Rd,\;Re,\;Rf\;and\;Rg_1$)을 고속액체크로마토그라피로 분석하는 일반적인 방법인 순상 column에서 $Rg_1$, Re 및 Rf가 명확히 분리되지 않는 문제점을 개선하기위하여 본 연구를 수행하였다. 고속액체크로마토그라피를 이웅하여 역상 ${\mu}{\beta}ondapak$ ODS컬럼으로 인삼중 주종 사포닌인 7종 ginsenosides $Rg_{1},\;Re,\;Rf,\;Rb_{1},\;RC,\;Rb_{2}$ 및 Rd를 양호하게 분리하였다. 이때 분석 조건으로 이동상 용매 조성은 (A) $H_{2}O$, (B) methyl cyanaide을 (A) 90/(B) 10에서 (A) 0/(B) 100으로 기울기 용리를 이용하였으며, 기울기 용리 제어장치를 사용하여 용리시켰다. 용매 흐름속도는 1.5ml/min, 검출기는 UV detector(203nm)이었다. 이 방법은 분리능과 재현성 및 회수율이 양호하므로, 앞으로 인삼중 ginsenosides 분석에 응용될 수 있을 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.204-210
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• 2001

Paclitaxel can be produced in high yield and with a high degree of purify from plant cell cultures of Taxus chinensis. The complete purification method was systematically established and described. This method was an efficient procedure for the purification of paclitaxel from crude paclitaxel, consisting or reverse-phase chromatography, followed by a normal-phase chromatography. The two-stage HPLC purification scheme serves as an effective and economical approach for resolving paclitaxel from complex mixtures of taxoids, with high purify (>99%) and low impurities (<0.1%). The process is readily scalable to a pilot plant and eventually to a production environment where multikilogram quantities of material are expected to be produced. The process has been optimized to minimize solvent usage, complexity, and operating costs.

• 분석과학
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• 제11권6호
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• pp.436-439
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• 1998

인삼 사포닌을 역상 고속 액체크로마토그래피 방법을 이용하여 여러 종류의 컬럼으로 분석하였다. 최적 조건은 Novapak $C_{18}$ ODS 역상 칼럼($3.9mm{\times}150mm$, $5{\mu}m$), $CH_3CN/H_2O$ 이성분 이동상 용액과 기울기 제어장치, 그리고 1.5 mL/min의 용매 흐름속도였으며, UV 검출기(203 nm)를 사용하였다. 인삼의 주성분인 ginsenoside $Rb_1$, $Rb_2$, Rc, Rd, Re, Rf 그리고 $Rg_1$ 7종을 50분 안에 양호하게 분리하였다. 각 성분에 대한 검량선은 직선성이 좋았으며, 회귀계수는 0.98~0.99이었다.

• 한국식품과학회지
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• 제29권3호
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• pp.412-416
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• 1997

7개의 국내 시판 사과 주스제품에 대해 patulin 오염수준을 분석하였다. Sep-Pak silica cartridge를 이용한 ethyl acetate 추출물의 정제과정은 HPLC 분석시 추출물내의 다른 성분으로부터 patulin의 분리를 용이하게 하였다. Patulin 검출은 J'sphere $4\;{\mu}m$ ODS-H80 column을 사용하여 reverse phase HPLC로 274 nm에서 분석하였다. 분석결과 4개의 제품은 $4{\sim}20\;{\mu}g/L$ 수준으로, 나머지 3개 제품은 $30{\sim}45\;{\mu}g/L$ 수준으로 patulin에 오염되어 있었다. 이 분석방법에서의 검출 한계농도는 $3\;{\mu}g/L$이며 $3.78{\sim}125.9\;{\mu}g/L$ 오염수준에 대해 $80{\sim}90%$의 회수율을 보였다.