• 분석과학
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• 제5권4호
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• pp.465-469
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• 1992

유당분해효소의 활성도를 정량하기 위하여 고속액체크로마토그래피(HPLC)를 사용하는 방법을 확립하였다. Aminex HPX-87C 컬럼 및 RI 검출기를 사용하여 유당, 글루코스 및 갈락토스를 12분 이내에 정량적으로 분석할 수 있었다. 컬럼 온도는 $85^{\circ}C$로 고정하였으며 이동상으로는 탈이온화된 증류수를 사용하였다. ONPG(ortho-nitrophenol-${\beta}$-D-galactopyranoside)를 이용하는 발색법의 결과와 비교하여, 고속액체크로마토그래피에 의한 유당분해효소 활성도 측정법의 타당성을 고찰하였으며, 그 결과 ONPG 방법과 고속액체크로마토그래피 방법에 의한 효소활성도 측정의 실험결과에는 큰 차이가 없었으며, 고속액체크로마토그래피를 사용하는 경우에는 420 nm에 대한 방해 작용이 있는 기질에서도 유당분해효소의 활성도를 측정할 수 있는 장점을 가짐을 확인하였다.

• 생약학회지
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• 제49권1호
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• pp.76-83
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• 2018

A traditional herbal formula, Gyeji-tang has been used to treat the early colds, headache, chills, and fever in Asian countries. In this study, we were performed simultaneous determination of the 14 bioactive marker compounds, gallic acid, spinosin, paeoniflorin, albiflorin, liquiritin apioside, liquiritin, 6'''-feruloylspinosin, liquilitigenin, coumarin, cinnmamic acid, benzoylpaeoniflorin, cinnamaldehyde, glycyrrhizin, and 6-gingerol in Gyeji-tang using an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS). Analytical column was used a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) and maintained at $45^{\circ}C$ with a flow rate of 0.3 mL/min. The mobile phase consists of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was conducted using multiple reaction monitoring in the positive and negative modes by a Waters ACQUITY TQD LC-MS/MS system. The calibration curves of 14 bioactive marker compounds showed linearity with correlation coefficients ${\geq}0.9798$. The limits of detection and quantification values were in the range of 0.11-6.66 ng/mL and 0.34-19.99 ng/mL, respectively. As a result of the analysis using the established LC-MS/MS method, the amounts of tested 14 compounds in the lyophilized Gyeji-tang sample were detected up to $85.7{\mu}g/g$. These results may be useful for quality assessment of a traditional herbal formulas.

• Natural Product Sciences
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• 제18권3호
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• pp.158-165
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• 2012

A UPLC method for the simultaneous determination of seven compounds was established for the quality control in H. cordata. The UPLC was performed on a $C_{18}$ HSS T3 $2.1{\times}100$ mm, 1.8 ${\mu}m$ column during a 13 minute gradient elution of 0.2% aqueous acetic acid and acetonitrile with the flow rate of 0.2 mL/min at $30^{\circ}C$. The UPLC method was validated according to the ICH guideline of analytical procedures with respect to precision, accuracy, and linearity. The limit of determination and quantitation for the seven compounds were 0.01-0.09 and 0.03-0.28 ${\mu}g/mL$, respectively. The calibration curves of all seven compounds showed good linearity ($r^2$ > 0.999). The intra-day and inter-day the RSD values used to evaluate the precision of analysis were less than 0.9%. The recoveries of quantified compounds ranged from 98.63 to 103.85%. The developed UPLC method was found to be effective, convenient and sensitivity for quantitative analysis of seven compounds in H. cordata. This work could be provided a baseline source for quality control of H. cordata.

• Mass Spectrometry Letters
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• 제10권1호
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• pp.18-26
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• 2019

We developed a bioanalytical method for simultaneous determination of nine NBOMe derivatives (25H-NBOMe, 25B-NBOMe, 25E-NBOMe, 25N-NBOMe, 25C-NBOH, 25I-NBOH, 25B-NBF, 25C-NBF, and 25I-NBF) in human plasma using liquid chromatography tandem mass spectrometry (LC-MS/MS). Human plasma samples were pre-treated using solid-phase extraction. Separation was achieved on a C18 column under gradient elution using a mobile phase containing 0.1% formic acid in acetonitrile and 0.1% formic acid in water at a flow rate of 0.3 mL/min. Mass detection was performed in the positive ion mode using multiple reaction monitoring. The calibration range was 1-100 ng/mL for all quantitative analytes, with a correlation coefficient greater than 0.99. The intra- and inter-day precision and accuracy varied from 0.85 to 6.92% and from 90.19 to 108.69%, respectively. The recovery ranged from 86.36 to 118.52%, and the matrix effects ranged from 27.09 to 99.72%. The stability was acceptable in various conditions. The LC-MS/MS method was validated for linearity, accuracy, precision, matrix effects, recovery and stability in accordance with the FDA guidance. The proposed method is suitable for reliable and robust routine screening and analysis of nine NBOMe derivatives in forensic field.

• Journal of Applied Biological Chemistry
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• 제59권2호
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• pp.165-169
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• 2016

The quantitative analytical method for the bioactive substance, 3-cyano-4-methoxy-N-methyl-2-pyridone (ricinine) and an index compound, ricinoleic acid in castor plant (Ricinus communis) extract or oil was developed. For the determination of a pyridone alkaloid compound, ricinine, successive cartridge cleanup method combined with ultra-performance liquid chromatography was set up with $ENVI-Carb^{TM}$ (0.5 g) and $C_{18}$ SPE cartridges. Accuracy and precision were evaluated through fortification studies of one biopesticide (PE) at 10 and $100mg\;kg^{-1}$. Mean recoveries of ricinine were 98.7 and 96.0 % associated with less than 10 % RSD, respectively. For the determination of ricinoleic acid in castor extract and oil, saponification and methylation were optimized using gas chromatography-time of flight mass spectrometry. Recovery was more than 84.8 % associated with 6.2 % RSD after derivatization procedure. Both methodologies developed were applied to analyze real samples including three castor oil products and six commercially available biopesticides containing R. communis, collected at Korean market. The contents of ricinine and ricinoleic acid in most commercial biopesticides were less than the oil or extract contents indicated by label.

• 생약학회지
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• 제40권1호
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• pp.18-24
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• 2009

GCSB-5 preparation is a purified extract from a mixture six herbal medicines (Acanthopanacis Cortex, Achyranthis Radix, Saposhnikoviae Radix, Cibotii Rhizoma, Glycine Semen Nigra, Eucommiae Cortex) that have been widely used in traditional medicine to treat various bone disorders. This study was carried out to obtain the HPLC analysis method that can be used to establish quantitative analysis of Glycine Semen Nigra and Eucommiae Cortex for standardization of GCSB-5 preparation. HPLC analysis methods for the simultaneous determination of genistin (Glycine Semen Nigra) and geniposide (Eucommiae Cortex) were established for the quality control of herbal medicinal raw material and preparation. And validation of HPLC analysis methods were conformed for verification of HPLC methods by check to specificity, linearity, intra-day precision, inter-day precision and accuracy following ICH guideline. As the result of quantitative analysis, the contents of genistin and geniposide in the raw material of GCSB-5 preparation were 0.0426-0.0427 mg/g and 0.431-0.432 mg/g. And GCSB-5 preparation contained genistin of 0.0202-0.0203 mg/capsule and geniposide of 0.211-0.212 mg/capsule, respectively.

• 한국수정란이식학회지
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• 제28권4호
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• pp.343-348
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• 2013

Cell-free fetal RNA has been highlighted as useful tools for the fetal sex determination or other genetic inherent disorder. However, there is no knowledge about the sex determination using cell free fetal RNA in bovine field. Thus, the present study aimed to evaluate the presence of transcripts of DDX3Y, USP9Y and ZRSR2Y genes in maternal plasma of pregnant cows to determine the sex of the fetus using real-time quantitative polymerase chain reaction assay, and verify its accuracy, sensitivity and specificity compared with the molecular testing and the calf sex at birth. Transcripts of USP9Y and DDX3Y genes were expressed in the all plasma of males and females both the control group and the experimental group. However, ZRSR2Y gene was matched up with the molecular testing and the true sex in control group and has an overall accuracy of 82.6%, a sensitivity of 75%, and a specificity of 100% in experimental group. Therefore, these results indicated that real time PCR technique, as a noninvasive and cost-efficient method, is possible to determination fetal sex in the bovine species using circulating cell free RNA in maternal plasma and especially ZRSR2Y gene could be a good candidate for the RNA based sex determination work.

• 대한화학회지
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• 제56권2호
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• pp.228-235
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• 2012

$N_2O_2$계 시프염기 리간드인 N,N'-bis(4-methoxysalicylidene)phenylendiamine(4-$CH_3O$-salphen)을 합성하였다. 합성한 4-$CH_3O$-salphen을 이용하여 분광광도법으로 수용액 중의 Cu(II)이온 정량실험을 시도하였다. Cu(II)이온 정량을 위한 최적 실험조건을 구한 결과, 4-$CH_3O$-salphen 농도는 $2.0{\times}10^{-4}\;mol/L$, 용매 DMSO와 물의 비율은 50/50(v/v), pH는 5.5에서, 온도는 $55^{\circ}C$에서 한 시간정도를 물중탕하고, 시료의 흡광도는 388 nm였고, 그 조건에서 검량곡선을 작성하였다. 작성된 검량곡선(${\varepsilon}=3.6{\times}10^4\;mol^{-1}cm^{-1}$)은 $R^2$=0.9963의 상관계수 값을 나타내었다. 이상의 최적화된 실험조건을 이용하여 온천수, 반도체 공장폐수 및 하수 처리장의 처리수를 채취하여 Cu(II)이온을 각각 정량 분석한 결과는 측정 평균값이 기준 값에 대하여 0.6~5.4% 범위에서 잘 일치 하였고, 정량한계는 31.77 ng/mL($5.0{\times}10^{-7}\;mol/L$)이었다.

• 대한의생명과학회지
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• 제18권2호
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• pp.96-103
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• 2012

Kv 4.2 ion channel protein has an ability to open at subthreshold membrane potentials and to recover quickly from inactivation. That is very important for neuronal signal transmission in vertebrate brain. In order to purify Kv 4.2 protein, the novel purification methods were experimented. The purification procedure utilized chromatography on DE-52 ion exchange column and affinity chromatography on a WGA-Sepharose 4B, and Kv 4.2 affinity column chromatography. It was found that 0.5% (wt./vol.) Triton X-100 detergent in lysis buffer worked well for Kv 4.2 protein solubilization from rat brain membrane. Protein quantitative determination was conducted by BCA method at 562 nm for each purification step to avoid determination interference of protein at 280 nm by detergent. The confirmation of Kv 4.2 existence and amount is performed using by SDS-PAGE/immunoblotting or 96-well dot blotting. The Kv 4.2 without interacting protein that contains carbohydrate, was purified from novel biochemical 3-steps purification method for further research.

• 생약학회지
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• 제41권1호
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• pp.62-66
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• 2010

In result of analysis for the content determination of ursolic acid(UA) and oleanolic acid(OA) in extract of Eriobotryae folium using HPLC with UV detector, UA in chloroform extract of Moo-mok variety was showed highest content(2.7843 mg/g). And OA in ethyl acetate extract of Dae-bang variety was showed highest content(0.5898 mg/g). These result suggest that direct extraction using organic solvent(chloroform or ethyl acetate) was useful method for rapid quantitative analysis of UA and OA without preprocessing such as drying or fractionation.