• 대한본초학회지
• /
• 제30권4호
• /
• pp.11-20
• /
• 2015

Objectives : Ongyeong-tang (OGT) is a traditional herbal formula used to cure gynaecological disorders. OGT consists of 12 herbal medicines containing various bioactive components. Therefore, the development of suitable analytical method for the marker compounds is necessary for the quality control of OGT. Methods : Determination of the 18 marker compounds in OGT preparations was quantitatively performed by high-performance liquid chromatography-photodiode array detection analysis. The marker compounds were separated on a reversed-phase C ₁₈ column and the analytical method was successfully validated, which was applied to compare OGT extracts from laboratory preparation and commercial OGT granules. Results : Limit of detection and limit of quantification values were in the ranges of $0.001-0.016{\mu}g/mL$ and $0.003-0.047{\mu}g/mL$, respectively. Precision was 0.03-3.71 % within a day and 0.03-3.81 % over four consecutive days. Recovery of marker compounds ranged from 90.63-108.26 %, with relative standard deviation (RSD) values < 4.0 %. Reproducibility was < 2.5 % of the RSD value. The 18 marker compounds were stable within 16 h at $10^{\circ}C$, with the RSD value < 3.5 %. Quantitative analysis results showed that the quantities of the 18 marker compounds varied among OGT samples. Pearson coefficient evaluation and principal component analysis demonstrated that an OGT water extract produced by a laboratory method clearly differed from commercial OGT granules. Conclusions : The developed analytical method was simple, precise, and reliable. Therefore, it can be used for the quality assessment of OGT preparations.

• Asian-Australasian Journal of Animal Sciences
• /
• 제31권8호
• /
• pp.1315-1324
• /
• 2018

Objective: An experiment was conducted to investigate the effects of different diet formulations: F1 (Two complicated basal diets containing different crude protein levels plus tested feedstuff) vs F2 (A simple corn soybean meal [SBM] basal diet plus tested feedstuff) combined with total collection (TC) or chromic oxide ($Cr_2O_3$) marker or acid-insoluble ash (AIA) marker method, and freeze-dry or oven-dry (OD) technique on estimation of nutrient digestibility in diets fed to growing pigs. Methods: In F1, twelve barrows were allocated to two $6{\times}4$ Youden Squares. The treatment diets included a high protein basal (HPB) diet, a low protein basal (LPB) diet, a corn diet and a wheat bran (WB) diet formulated based on the HPB diet, and a SBM diet and a rapeseed meal (RSM) diet formulated based on the LPB diet. In F2, eight barrows were allocated to two $4{\times}4$ Latin Squares. The treatment diets included a corn basal diet, a SBM basal diet formulated based on the corn diet, and a WB diet and a RSM diet formulated based on the SBM diet. Results: Concentration of digestible (DE) and metabolizable energy (ME), and the apparent total tract digestibility of gross energy, ash, neutral detergent fibre, and acid detergent fibre determined by $Cr_2O_3$ marker method were greater than those determined by TC and AIA marker methods in HPB, LPB, and RSM diets formulated by F1 and in corn diet formulated by F2 (p<0.05). The DE values in WB and both DE and ME values in SBM and RSM estimated using F1 were greater than those estimated using F2 (p<0.05). Conclusion: From the accuracy aspect, the AIA marker or TC method combined with OD technique is recommended for determining the energy concentration and nutrient digestibility of components in diets fed to growing pigs.

• 생약학회지
• /
• 제48권4호
• /
• pp.320-328
• /
• 2017

Yongdamsagan-tang has been used to treat the urinary disorders, acute- and chronic-urethritis, and cystitis in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was established for simultaneous analysis of the 20 bioactive marker compounds, geniposidic acid, chlorogenic acid, geniposide, liquiritin apioside, acteoside, calceolarioside B, liquiritin, nodakenin, baicalin, liquiritigenin, wogonoside, baicalein, glycyrrhizin, wogonin, glycyrrhizin, wogonin, saikosaponin A, decursin, decursinol angelate, alisol B, alisol B acetate, and pachymic acid in traditional herbal formula, Yongdamsagan-tang. Chromatographic separations of all marker compounds were conducted using a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was performed using a Waters ACQUITY TQD LC-MS/MS coupled with an electrospray ionization source in the positive and negative modes. The flow rate was 0.3 mL/min and injection volume was $2.0{\mu}L$. The correlation coefficient of 20 marker compounds in the test ranges was 0.9943-1.0000. The limits of detection and quantification values of the all marker components were 0.11-6.66 and 0.34-19.99 ng/mL, respectively. As a result of the analysis using the optimized LC-ESI-MS/MS method, three compounds, geniposidic acid (from Plantaginis Semen), alisol B (from Alismatis Rhizoma), and pachymic acid (from Poria Sclerotium), were not detected in this sample. While the amounts of the 17 compounds except for the geniposidic acid, alisol B, and pachymic acid were $0.04-548.13{\mu}g/g$ in Yongdamsagan-tang sample. Among these compounds, baicalin, bioactive marker compound of Scutellariae Radix, was detected at the highest amount as a $548.13{\mu}g/g$.

• 생약학회지
• /
• 제46권2호
• /
• pp.123-132
• /
• 2015

In this study, we carried out quantification analysis of the six marker components, geniposidic acid, chlorogenic acid, geniposide, pinoresinol diglucoside, liriodendrin, and genipin in the 70% ethanol extracts of non-processed Eucommiae Cortex and processed Eucommiae Cortex using a high-performance liquid chromatography coupled with photodiode array detector. The six components were separated on Gemini C₁₈ column (5 μm, 4.6×250 mm) by the gradient elution with 1.0% (v/v) acetic acid in water and 1.0% (v/v) acetic acid in acetonitrile as mobile phase. The flow rate was 1.0 mL/min and the injection volume was 10 mL. The amount of geniposidic acid, chlorogenic acid, geniposide, pinoresinol diglucoside, liriodendrin, and genipin in non-processed Eucommiae Cortex were 1.31, 0.31, 0.66, 0.46, 0.46, and 0.03%, respectively, while the amount of the six compounds in non-processed Eucommiae Cortex were 0.04-0.78, 0.01-0.14%, 0.05-0.63%, 0.01-0.37%, 0.15-0.42%, and not detected, respectively. After processing treatment, the contents of three iridoids, two lingnan, and one phenylpropanoid decreased in Eucommiae Cortex.

• 한국균학회지
• /
• 제20권4호
• /
• pp.315-323
• /
• 1992

재현성 있는 RAED marker들의 증폭을 위해서 PCR에 영향을 주는 조건들에 대해 조사를 하였다. 그 결과 약 15 ng의 DNA가 효과적인 PCR에 적합한 양이었으며 PCR에 사용되는 성분(reaction component)들의 농도가 PCR 결과에 있어서 상호의존관계에 있었고 DNA 용액에 포함되어 있는 RNA가 DNA 증폭을 방해하는 작용을 하였다. $25\;{\mu}l$의 PCR 반응용액에 30ng의 10-mer primer, $200\;{\mu}M$ dNTP, 0.001% gelatin, 1.5 mM $MgCl_2$, 10 mM Tris-Cl(pH 8.8), 50 mM KCl, 0.1%, Triton X-100, 2units의 Taq DNA polymerase, 그리고 RNA를 제거한 15 ng의 DNA를 사용한 결과 가장 재현성있는 RAPD marker들이 증폭되었다.

• 한국식품위생안전성학회지
• /
• 제37권1호
• /
• pp.29-37
• /
• 2022

본 연구에서는 발효노니를 건강기능식품 소재로 활용 시 기초자료로 제공하고자 발효노니 추출물 및 지표성분의 면역활성 증진 효과를 평가하였다. RAW 264.7 대식세포에서 발효노니 추출물 및 지표성분 6종을 처리하여 XTT 세포독성평가, Nitric Oxide 생성 측정, Cyokine 생성 측정, immune marker genes 발현분석 수행하였다. 뿐만 아니라 양성대조군으로 LPS와 기능성 원료로 사용되고 있는 발효홍삼 추출물을 사용하였다. 그 결과 모든 처리 농도 및 처리군에서 세포독성이 관찰되지 않았으며, 지표성분 6종 중 SCP 및 ASE에서 NO 생성이 증가됨을 확인하였다. 뿐만 아니라 ASE 처리군에서는 IL-6 및 IL-1β의 생성이 증가되었으며, iNOS 및 TNF-α의 immune marker genes 발현이 증가됨을 확인하였다. 발효노니 추출물 효능 평가에서는 발효시 NO 생성 및 IL-6, IL-1β의 생성, COX2의 발현이 증가되는 것으로 나타났다. 이러한 연구 결과는 발효노니 추출물 및 지표성분의 선천면역 활성증가를 타나내며 노니 및 발효노니 표준화 연구에서 지표성분으로 사용 가능성을 제시한다. 따라서 발효노니는 면역증진 활성을 갖는 제품 개발에 있어서 유용한 기능성 식품소재로써 사용될 수 있으며, 우수한 효능을 나타내는 지표성분은 유용성분으로 이용이 가능할 것으로 사료된다.

• 생약학회지
• /
• 제46권4호
• /
• pp.352-364
• /
• 2015

The aim of this study was to quantitatively analyze for quality assessment of eighteen marker compounds, including homogentisic acid, 3,4-dihydroxybenzaldehyde, spinosin, liquiritin, hesperidin, ginsenoside Rg1, liquiritigenin, ginsenoside Rb1, glycyrrhizin, 6-gingerol, atractylenolide III, honokiol, costunolide, dehydrocostuslactone, atractylenolide II, nootkatone, magnolol, and atractylenolide I, in Hyangsayukgunja-tang using an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer. The column for separation of eighteen marker components were used a UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}$) and kept at $45^{\circ}C$ by gradient elution with 0.1% (v/v) formic acid in water and acetonitrile as mobile phase. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}l$, respectively. The correlation coefficient of all marker compounds was ${\geq}0.9914$, which means good linearity, within the test ranges. The limits of detection and quantification values of the all analytes were in the ranges 0.04-1.11 and 0.13-3.33 ng/mL, respectively. As a result, five compounds, homogentisic acid, 3,4-dihydroxybenzaldehyde, spinosin, liquiritigenin, and atractylenolide I, in this sample were not detected and the amounts of the 13 compounds except for the 5 compounds were $8.10-6736.37{\mu}g/g$ in Hyangsayukgunja-tang extract.

• Journal of Microbiology and Biotechnology
• /
• 제14권4호
• /
• pp.783-788
• /
• 2004

Marker proteins of genetically-modified organisms (GMO) and their antibodies were prepared and characterized as major components of an analytical system. We selected two GMO markers, neomycin phosphotransferase II and 5- enolpyruvylshikimate-3-phosphate synthase, and produced them from E. coli employing genetic recombination technology. After purification, their structural conformation and binding affinities to the respective antibodies were characterized. The results showed that the recombinant proteins were identical with commercially obtained reference proteins. We further used them as immunogens to raise polyclonal antibodies capable of discriminating GMO containing protein from non-GMO. Well-characterized marker proteins and antibodies will be valuable as immunoreagents in constructing analytical systems such as biosensors and biochips to measure quantities of GMO.

• Journal of Plant Biotechnology
• /
• 제36권2호
• /
• pp.106-114
• /
• 2009

Genetic components introduced into approved GM crops are a key subject for safety assessment and provide a basis for the development of detection methods for GM crops. In order to understand the genetic components in approved GM crops comprehensively, we screened the genetic vector maps of GM crops that had been approved for commercialization around the world. A total of 64 varieties from 5 major GM crop species (maize, canola, cotton, soybean, and tomato) were subjected to analysis. The genetic components included genes, promoters, terminators, and selection marker. This survey may be useful for researchers who develop GM crops and methods for detecting GM crops.

• 약학회지
• /
• 제58권3호
• /
• pp.158-164
• /
• 2014

Dangguisu-san is a well-known traditional Korean herbal medicine prescription and has been widely used to treat ecchymosis, blood stagnation, and pain resulting from physical shock in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer method was established for the simultaneous determination of the 17 biomarker components in Dangguisu-san. All analytes were separated on an UPLC BEH $C_{18}$ ($100{\times}2.1$ mm, $1.7{\mu}m$) column and maintained at $45^{\circ}C$. The mobile phase consisted of two solvent systems, 0.1% (v/v) formic acid in water (A) and acetonitrile (B) by gradient flow. The injection volume was $2.0{\mu}l$ and the flow rate was 0.3 ml/min with detection at mass spectrometer. Calibration curves of the 17 biomarker components were acquired with $r^2$ values ${\geq}0.9951$. The values of limit of detection and quantification of all analytes were 0.02~6.32 ng/ml and 0.05~18.95 ng/ml, respectively. The amounts of the 17 components in Dangguisu-san sample were $3.17{\sim}13,224.50{\mu}g/g$.