• Journal of Applied Biological Chemistry
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• 제64권1호
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• pp.57-61
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• 2021

Saikosaponin derivatives such as saikosaponins A, B1, B2, B3, B4, C, and D present in Bupleurum falcatum were analyzed by a high performance liquid chromatograph equipped with an evaporative light scattering detector, using different extraction solvents (water and 70% ethanol). The samples were injected into a YMC Pack Pro C18 column and separated using a gradient elution system with a mobile phase composed of acetonitrile and water at a flow rate of 1.1 mL/min. The content of saikosaponin derivatives was higher in 70% ethanol extract than in water extract. This study provides an efficient analytical method for determining the optimal conditions for extraction of saikosaponin derivatives, which can be used as a basis for development of functional foods and pharmaceutical products from B. falcatum.

• Journal of Applied Biological Chemistry
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• 제64권1호
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• pp.13-17
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• 2021

Qantitative analysis of six compounds: (+)-catechin, benzoic acid, gallic acid methyl ester, paeonol, paeoniflorin, and albiflorin from Paeonia lactiflora extracts was performed using high-performance liquid chromatography and an ultraviolet (UV) detector, following different extraction methods. A reverse-phase column was used in a gradient elution system, and UV detection was performed at 280 nm. The results showed that the quantity of paeoniflorin was the highest in ethanol and water extracts (73.89 and 57.87 mg/g, respectively) among the six compounds. This study contributes a good analysis method for the contents of P. lactiflora and would be propitious for developing medicines and functional foods.

• 한국물환경학회지
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• 제25권5호
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• pp.720-726
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• 2009

We investigated the most effective pre-treatment processes and LC/MS/MS condition for microcystins analysis. With a step-by-step pre-treatment, efficiencies of several established methods were compared. At the level of cell burst, sonication method was found to be the most efficient. As a mycrocystins first extraction solvent, 5% acetic acid showed the highest efficiency. An isolation and recovery rate of mycrocystins of ODS Sep-Pak $C_{18}$ cartridge was higher than HLB SPE cartridge. As a final elution solvent from cartridge, 100% MeOH had a better efficiency than others. Using a LC/MS/MS, effective analytical methods were established. C18 reverse column was used and gradient elution was performed with using acetonitrile, 0.1% formic acid as a mobile phase. We analysed to 0.8 mL/min flow rate fit to the $5{\mu}m$ particle size column and $55^{\circ}C$ housing temperature. The validity of established analytical method was evaluated that MDL as average $0.050{\pm}0.014{\mu}g/L$ and LOQ as average $0.160{\pm}0.045{\mu}g/L$ had a good sensitivity over 40 magnification rather than $2{\mu}g/L$ detection limit of HPLC.

• 분석과학
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• 제19권6호
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• pp.519-528
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• 2006

본 연구에서는 뇨시료를 고체상 추출법으로 추출한 후 HPLC/DAD/ESI-MS(high-performance liquid chromatograph/diode array detector/electro-spray ionitation mass spectrometry)를 사용하여 분석하였다. 7종의 thyroid hormones의 HPLC 분리조건은 Hypersil ODS(octadecylsilica) 컬럼(4.6mm I.D., 100 mm length, particle size $5{\mu}m$)을 사용하고 ammonium formate buffer와 acetonitrile을 이동상으로 하여 기울기 용리한 결과 완전 분리가 가능하였으며, UV spectra 및 질량스펙트럼을 확인할 수 있었다. 고체상 추출법에 의한 전처리 최적 조건을 조사한 결과 시료를 pH 3으로 한 후 C18 고체상을 사용하여 4 mL의 methanol/ammonium hydroxide(9:1) 혼합용액으로 용리할 경우 회수율이 89.0-113.1%로 나타났다. HPLC/DAD를 이용하여 10-1000 ng/mL범위에서 검량선을 작성한 결과 $r^2$값은 0.992-0.998 으로 나타났으며 검출한계는 2-4 ng/mL(3.8-13.0 pmol/mL)로 계산되었다.

• 한국식품과학회지
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• 제50권4호
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• pp.377-382
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• 2018

상지추출물을 식약처 개별인정형 건강기능식품 기능성원료로 개발하기 위한 연구의 일환으로 상지추출물의 표준화를 위한 기능성지표성분 ORT 분석법 설정과 분석법에 대한 확인을 실시한 결과는 다음과 같다. 먼저 HPLC를 이용하여 0.05% 인산이 첨가된 물과 아세토나이트릴을 이동상으로 하고 YMC-Pack Pro $C_{18}$ column ($5{\mu}m$, $4.6{\times}250mm$)을 사용하여 기울기용출(gradient elution)으로 ORT 분석법을 확립한 후 분석법에 대하여 특이성, 직선성, 정확성, 회수율과 정밀성을 각각 측정하였다. ORT 검량선의 상관계수($R^2=1$)은 좋은 선형성을 보였으며 LOQ와 LOD는 각각 1.0와 $0.3{\mu}g/mL$ 나타났다. 정확도는 일내분석과 일간분석에서 각각 99.44-103.47%, 97.60-105.81%로 나타내었으며, 상대표준편차(RSD)는 각각 3.52, 4.70% 이었다. 회수율은 97.64-103.69% 이었으며, RSD는 0.89-1.14%으로 나타나서 RSD 2.00% 이하로 정밀성이 있음을 확인할 수 있었다. 위의 결과로부터 HPLC를 이용한 ORT의 분석법은 상지추출물의 기능성원료 표준화를 위한 지표성분 분석법으로 적합함을 확인하였다. 본 시험법에 따라 3회 제조한 상지추출물의 ORT 함량은 평균 3.74%로 나타내었다. 따라서 본 연구를 통하여 확립된 ORT의 분석법이 향후 상지추출물의 기능성원료 표준화와 규격화를 위한 기초자료로서 크게 활용될 것으로 기대된다.

• Mass Spectrometry Letters
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• 제4권3호
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• pp.47-50
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• 2013

An ultra-fast generic LC-MS/MS method was developed for high-throughput quantification of discovery pharmacokinetic (PK) samples and its reliability was verified. The method involves a simple protein precipitation for sample preparation and the analysis by ultra-fast generic LC-MS/MS with the ballistic gradient program and selected reaction monitoring (SRM) mode. Approximately 290 new chemical entities (NCEs) (over 10,000 samples) from 5 therapeutic programs were analyzed. The calibration curves showed good linearity in the concentration range of 1, 2 or 5 to 2000 ng/mL. No significant ion suppression was observed in the elution region of all the NCEs. When approximately 300 plasma samples were continuously analyzed, the peak area of internal standard was constant and reproducible. In the repeated analysis of samples, the plasma concentrations and the area under the curve (AUC) were consistent with the results from the first analysis. These results showed that the present ultra-fast generic LC-MS/MS method is reliable in terms of selectivity, sensitivity, and reproducibility and could be useful for high-throughput quantification and other bioanalysis in drug discovery.

• 미생물학회지
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• 제21권4호
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• pp.221-228
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• 1983

Saccharomyces cerevisiae로 부터 glucose-6-phosphate dehydrogenase을 신속하고 간편하게 정제하는 과정을 affinity chromatography를 이용하여 개발하였다. 이 효소를 정제하는데 적절한 affinity medium을 조사해 본 결과, $NADP^+ -agarose$와 Affi-gel Blue(Cibacron Blue F3GA)가 Affi-gel Red(Procion Red HE-3B), AMP-agarose, ATP-agarose, 그리고 $NADP^+ -agarose$보다 유용함이 밝혀졌다. 이 두가지 affinity media에 흡착된 효소를 분리 하는데 가장 적합한 elution 조건을 조사하였는데 KCI gradient( (0-1.OM)가 효소의 순도 및 수회율을 가장 높일 수 있는 적합한 방법이었다. 특히 Affi-gel Blue를 사용할 경우, KCI gradient로 효소를 용출시키기 전에 NAD-(15mM)로NAD+에 친화역을 갖는 효소들을 제거하는 것이 enzyme의 순도를 높이는데 매우 효과적이었다. 그 결과 glucose-6-phosphate dehydrogenase를 baker's yeast로 부터 기존의 간단한 정제 과정과 affinity chromatography를 병행한 방식을 샤용하여 분리 하였는데, affinity medium으로 Affi-gel Blue를 사용했을 때는 180배 정도, NADP+-agarose를 사용했을 때는 2,000배 정도로 정제 되었다. 대량으로 glucose-6-phosphate dehydrogenase를 정제하는 경우, Affi-gel Blue를 사용하던 효소의 순도는 NADP+-agarose보다 낮으나, 효소의 회수율은 훨씬 더 높았다. 또한 G-6-P dehydrogenase에 대한 affinity medium의 capacity도 Affi-gel Blue가 NADP+-agarose보다 5배정도 높았으며 더우기 Affi-gel Blue는 여러번 반복적으로 사용될 수 있고, 그 제조 과정도 NADP+-agarose보다 간단하며 경비도 적게 들었다.

• Journal of Microbiology and Biotechnology
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• 제7권5호
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• pp.310-317
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• 1997

Enzymatic synthesis of glucosyl-sugar alcohols using various transglycosylating enzymes, such as cyclodextrin glucanotransferase (CGTase), ${\alpha}$-amylase, ${\alpha}$-glucosidase, and pullulanase was investigated using various sugar alcohols, such as sorbitol, xylitol, inositol, maltitol, and lactitol as glucosyl acceptors. CGTase showed the highest transglycosylating activity to sugar alcohols compared to other transglycosylating enzymes, and inositol and maltitol were the most suitable glucosyl acceptors. Soluble starch, extruded starch, cyclodextrins, and maltooligosaccharides were also identified to be adequate glucosyl donors for transglycosylation reaction of CGTase to sugar alcohols. The synthesis of glucosyl-maltitol in the reaction system using extruded starch as the glucosyl donor and maltitol as the glucosyl acceptor showed the best results showing the highest transglycosylation yield. The transglycosylation products were purified by activated carbon column chromatography with ethanol gradient elution. Chemical structures of above transglucosylated products were analyzed by nuclear magnetic resonance spectroscopy, and two products were identified to be maltotritol and maltotetraitol, in which one or two glucose molecules attached to the parent maltitol molecule by a ${\alpha}$-l,4-glucosidic bond, respectively.

• Food Science and Biotechnology
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• 제17권2호
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• pp.430-433
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• 2008

A high performance liquid chromatography (HPLC) method has been successfully developed to identify and quantify major ginsenosides in Korean ginseng seeds and roots. Using gradient elution of acetonitrile and water without buffer, the 6 major ginsenosides ($Rb_1,\;Rb_2$, Rc, Rd, Re, and $Rg_1$) were identified. Compared with ginseng roots, the amount of ginsenoside Re and Rd in ginseng seeds were significantly higher than those in ginseng roots (p<0.05). In ginseng seeds, the content of protopanaxtriol (PPT) was higher than that of protopanaxdiol (PPD) and the ratio of PPT and PPD was approximately 2.2 : 1. However, the content of PPT was lower than that of PPD in ginseng roots. It should be mentioned that both content of PPT and PPD in ginseng seeds were much higher than those in ginseng roots.

• 혜화의학회지
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• 제23권1호
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• pp.93-104
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• 2014

Glycyrrhizae Radix et Rhizoma has been extensively used by human beings as a medicinal herb as well as natural sweetener. In this study, we performed quantification analysis of three major constituents including liquiritin, glycyrrhizin, and glycyrrhetinic acid in the 70% ethanol extracts of non-processed Glycyrrhizae Radix et Rhizoma and processed Glycyrrhizae Radix et Rhizoma using a high-performance liquid chromatography coupled with photodiode array detector. The analytical column for separation of the 3 constituents used a Gemini C18 column kept at $40^{\circ}C$ by the gradient elution of two mobile phase. The amounts of liquiritin, glycyrrhizin, and glycyrrhetinic acid in non-processed Glycyrrhizae Radix et Rhizoma were 2.57%, 3.52%, and not detected. After processing by roasting, the best roasting temperature and time of iquiritin, glycyrrhizin, and glycyrrhetinic acid were $160^{\circ}C$-15 min (2.46%), $160^{\circ}C$-15 min (3.67%), and $240^{\circ}C$-15 min (0.76%), respectively.