• 대한한의학방제학회지
• /
• 제21권1호
• /
• pp.177-185
• /
• 2013

Objectives : Leejung-tang (Lizhong-tang) has been used for treatment of gastrointestinal disorders in Korea. In this study, we performed quantification analysis of five marker components, liquiritin, ginsenoside Rb1, ginsenoside Rg1, glycyrrhizin, and 6-gingerol in Leejung-tang using a ultra performance liquid chromatography- electrospray ionization-mass spectrometer (UPLC-ESI-MS). In addition, we evaluated antioxidant activity of Leejung- tang. Methods : The column for separation of five constituents used a UPLC BEH C18 ($100{\times}2.1mm$, $1.7{\mu}m$) maintained at $45^{\circ}C$. The mobile phase consisted of two solvent systems, 0.1% (v/v) formic acid in H2O (A) and CH3CN (B) by gradient flow. The flow rate was 0.3 mL/min with detection at mass spectrometer. The antioxidative activities conduct an experiment on 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of Leejung-tang. Results : Calibration curves of five marker compounds were acquired with r2 values > 0.99. The amount of the five compounds in Leejung-tang were 0.07 - 0.84 mg/g. The concentration required for 50% reduction (RC50) against ABTS radical was 119.02 ug/mL. In addition, the scavenging against DPPH radical of Leejung-tang was 11.4%, 14.5%, 19.8%, 29.6%, and 49.2% at 25 ug/mL, $50{\mu}g/mL$, $100{\mu}g/mL$, $200{\mu}g/mL$, and $400{\mu}g/mL$, respectively. Conclusions : The established LC-MS/MS method will be helpful to improve quality control of Leejung-tang. In addition, Leejung-tang is a potential antioxidant therapeutic agent.

• Mass Spectrometry Letters
• /
• 제6권4호
• /
• pp.116-119
• /
• 2015

Globotriaosylsphingosine (lyso-Gb3) is considered as one of the biological marker for Fabry disease. To date, a reliable biomarker that reflects disease severity and progression has not been discovered to guide the management of Fabry disease. A new method included a simple protein precipitation with acetonitrile in 100 μL of plasma following analyte separation on an Phenomenex Kintex- C18 column using a gradient elution (0.1% formic acid in 5-90% acetonitrile). Total run time was within 12 min including sample preparation and MS/MS analysis. The limit of detection and limit of quantitation were 1 ng/mL and 2 ng/mL, respectively. The calibration curve was linear over the concentration range of 2.0-200.0 ng/mL (r² = 0.9999). Inter-day accuracy and precision at 7 level were 93.4-100.7% with RSD of 0.55-5.97%. Absolute recovery was 97.6-98.6%. The method was applied to human and mice plasma, proved the suitability for quantification of lyso-Gb3 for screening, diagnosis and therapeutic monitoring of Fabry disease patients.

• Natural Product Sciences
• /
• 제21권1호
• /
• pp.20-24
• /
• 2015

Cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) were isolated by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water/trifluoroacetic acid (1 : 3 : 1 : 5 : 0.01, v/v) to give pure C3G (34.1 mg) and C3R (14.3 mg) from 1.5 g crude mulberry fruit extract. Using the pure C3G and C3R, a reliable high-performance liquid chromatography (HPLC) method was developed and validated to determine the C3G and C3R contents in mulberry fruit. C3G and C3R were separated simultaneously using an Eclipse XDB-C18 column ($4.6{\times}250mm$ I.D., $5{\mu}m$) coupled with a photodiode array detector (PDA). The gradient elution of the mobile phase consisting of acetonitrile (0.5% formic acid) and water (0.5% formic acid) was applied (1.0 mL/min), and the detection wavelength was 520 nm. The calibration curves of C3G and C3R showed good linearity (both with $r^2=0.9996$) in the concentration range $15.625-500{\mu}g/mL$, and the relative standard deviations (RSD%) of intra- and inter-day variability were in the ranges 2.1 - 8.2% and 4.1 - 17.1%, respectively. The accuracies were ranged 96.5 - 102.6% for C3G and C3R, respectively. The developed HPLC method was used to determine the contents of C3G and C3R in newly harvested mulberry from eight different provinces of Korea.

• Bulletin of the Korean Chemical Society
• /
• 제34권6호
• /
• pp.1684-1688
• /
• 2013

A rapid and sensitive method for determining usnic acid of Lethariella cladonioides in rat was established using high performance liquid chromatography (HPLC) quadrupole time-of-flight (QTOF) tandem mass (MS/MS). Rat plasma was pretreated by mixture of acetonitrile and chloroform to precipitate plasma proteins. Chromatographic separation was achieved on a column ($50{\times}2.1$ mm, $5{\mu}m$) with a mobile phase consisting of water (containing $5{\times}10^{-3}$ M ammonium formate, pH was adjusted to 3.0 with formic acid) and acetonitrile (20:80, v/v) at a flow rate of 0.3 mL/min. A tandem mass spectrometric detection with an electrospray ionization (ESI) interface was conducted via collision induced dissociation (CID) under negative ionization mode. The MS/MS transitions monitored were m/z 343.0448 ${\rightarrow}$ m/z 313.2017 for usnic acid and m/z 153.1024 ${\rightarrow}$ m/z 136.2136 for protocatechuic acid (internal standard). The linear range was calculated to be 2.0-160.0 ng/mL with a detection limit of 3.0 pg/mL. The inter- and intra-day accuracy and precision were within ${\pm}7.0%$. Pharmacokinetic study showed that the apartment of usnic acid in vivo confirmed to be a two compartment open model. The method was fully valid and will probably be an alternative for pharmacokinetic study of usnic acid.

• 한국환경농학회지
• /
• 제35권3호
• /
• pp.166-174
• /
• 2016

농경지 토양에서 대상항생제 6종(amoxicillin, ampicillin, chlortetracycline, enrofloxacin, oxytetracycline, tetracycline을 아세트산 함유 아세토니트릴, Na₂Cit.5H₂O, Na₃Cit.2H₂O과 Na₂-EDTA로 추출 후 C₁₈으로 정제하여 LC-MS/MS로 분석하는 모니터링을 수행하였다. 그 중 2종의 항생제, chlortetracycline과 enrofloxacin이 국내 농경지 토양에서 검출되었다. 제안한 분석법은 토양 중 잔류 항생제 모니터링을 위한 빠르고 간편한 시험법이었고 다양한 활용이 가능할 것으로 보인다.

• 분석과학
• /
• 제8권4호
• /
• pp.683-689
• /
• 1995

Electrochemical reduction of carbon dioxide under high pressure on Fe electrodes and a gas diffusion electrode containing Pt catalyst (Pt-GDE) had been investigated. Formic acid was formed on Fe electrode with a faradaic efficiency of 60% at a current density of $120mA\;cm^{-2}$ under 30 atm of $CO_2$. Hydrocarbons such as $CH_4$, $C_2H_6$, $C_3H_6$, $1-C_4H_8$, and $n-C_5H_{12}$ are also formed. The distribution of hydrocarbons followed well the Schultz-Flory distribution. $CH_4$ was formed efficiently as the main reduction product on Pt-GDE.

• 한국식품영양과학회지
• /
• 제41권11호
• /
• pp.1554-1558
• /
• 2012

고창 복분자의 개별인정형 건강기능식품 기능성 원료로 개발하기 위하여 지표성분 표준화를 위한 ellagic acid의 분석법 설정과 분석법에 대한 밸리데이션을 실시하였다. 1% formic acid가 첨가된 water와 acetonitrile을 이동상으로 하고 Symmetry$^{(R)}$ (C18, $4.6{\times}250mm$, $5.0{\mu}m$) 칼럼을 사용하여 기울기용리(gradient elution) 방법으로 분석하였다. 본 연구에서는 분석법을 확립하고 분석법에 대하여 특이성, 직선성, 정확성과 정밀성 그리고 회수율에 대하여 확인하였다. Ellagic acid의 검량선은 $R^2=0.9996$으로 좋은 선형성을 보였으며 LLOQ와 LOD는 각각 $1{\mu}g/mL$와 $0.3{\mu}g/mL$였다. 일내와 일간 분석에서 상대표준편차(RSD)는 각각 2.28%와 2.84%미만으로 나왔다. 회수율 측정결과에서는 89.17~97.92%로 나왔고, RSD는 0.05~0.14%였다. 그러므로 HPLC를 이용한 ellagic acid의 분석법이 고창 복분자 추출물의 기능성원료 표준화를 위한 지표성분 분석을 위한 적합한 시험법임이 검증되었다. 본 시험법에 따라 분석한 고창 미숙과 복분자 추출물 내의 ellagic acid의 함량은 세 lot를 3번씩 분석하였을 때 약 $1.92{\mu}g/mg$(0.192%)이 나왔으며 RSD값은 2.36% 이하였다. 따라서 본 연구를 통하여 확립된 ellagic acid의 분석법이 고창 복분자의 개별인정형 건강기능식품 기능성 원료 개발을 위한 기초자료로 활용될 것으로 본다.

• 한국작물학회지
• /
• 제53권4호
• /
• pp.359-368
• /
• 2008

검정콩 품질평가의 한 기준이 되는 안토시아닌의 함량평가를 위한 분석기술 개발의 일환으로 검정콩 종피 함유 안토시아닌의 최적 HPLC 분석조건을 확립하여 검정콩 함유 안토시아닌의 작물학적 연구와 검정콩 함유 안토시아닌의 이용도 증진 및 상업적 활용에 기초 자료를 제공하고자, 검정콩에 함유된 3종의 안토시아닌 성분을 동시에 분석할 수 있고, 재현성 및 검출 감도가 증진된 최적의 HPLC 분석 조건을 검토한 결과를 요약하면 다음과 같다. 1. 검정콩에 함유된 3종의 안토시아닌 D3G, C3G 및 Pt3G를 동시에 분석할 수 있는 최적의 HPLC 분석 조건을 검토한 결과 컬럼은 YMC-pak ODS-AM 303($4.6{\times}250\;mm$, $5\;{\mu}m$) 컬럼을 이용하고, 검출파장은 520 nm, 컬럼온도를 $30^{\circ}C$, 분당유속은 0.7 mL, 이동상 용매로는 A용매로 5% 포믹산 - 증류수, B용매로 5% 포믹산 - 아세토니트릴을 사용하여 농도구배 조건으로 분석하는 것이 가장 분리능이 향상된 효율적 분석법으로 조사되었다. 2. 검정콩에 함유된 3종의 안토시아닌 D3G, C3G 및 Pt3G를 동시에 분석할 수 있는 최적 HPLC 분석 조건의 재현성 검정을 실시한 결과 머무름시간(Rt.)의 변이계수는 최대 1.24%, peak height의 변이계수는 최대 2.04%, peak area의 변이계수는 최대 0.21% 미만을 나타내므로 확립된 검정콩 함유 안토시아닌 동시분석을 위한 최적 HPLC 분석 조건은 극히 높은 분석 재현성을 나타내었다. 3. 확립된 최적 HPLC 분석조건의 최소 검출한계(LOD, Limit of Detection)를 평가하기 위해 검출기의 signal/noise ratio = 3인 조건에서 최소 검출한계를 조사한 결과 3종의 검정콩 종피 함유 안토시아닌 색소의 최소 검출한계는 최소 10 ppb(0.01 ppm, 10 ng/mL) 이하에서도 완벽하게 검출되는 양상을 나타내므로 확립된 최적의 HPLC 분석조건은 검정콩 함유 안토시아닌의 작물학적 특성 평가용으로 활용하기에 전혀 무리가 없는 분석법으로 조사되었다.

• 한국지하수토양환경학회지:지하수토양환경
• /
• 제18권1호
• /
• pp.67-77
• /
• 2013

To improve the energy efficiency of conventional thermal treatment, soil remediation with microwave has been studied. In this study, the remediation efficiency of contaminated soil with semi-volatile organic contaminants were evaluated with microwave oven and several additives such as water, formic acid, iron powder, sodium hydroxide (NaOH) solution, and activated carbon. For the experiment, loamy sand and sandy loam collected from Imjin river flood plain were intentionally contaminated with hexachlorobenzene and phenanthrene, respectively. The contaminated soils were treated with microwave facility and the mass removals of organic contaminants from soils were evaluated. Among additives that were added to increase the remediation efficiency, activated carbon and NaOH solution were more effective than water, iron powder, and formic acid. When 10 g of hexachlorobenzene (142.4 mg/kg-soil) or phenanthrene (2,138.8 mg/kg-soil) contaminated soil that mixed with 0.5 g iron powder, 0.5 g activated carbon and 1 ml 6.25 M NaOH solution were treated with microwave for 3 minutes, more than 95% of contaminants were removed. The degradation of hexachlorobenzene during microwave treatments with additives was confirmed by the detection of pentachlorobenzene and tetrachlorobenzene. Naphthalene and phenol were also detected as degradation products of phenanthrene during microwave treatment with additives. The results showed that adding a suitable amount of additives for microwave treatments fairly increased the efficiency of removing semi-volatile soil organic contaminants.

• Applied Biological Chemistry
• /
• 제13권1호
• /
• pp.65-72
• /
• 1970

Bence Jones 단백질중(蛋白質中) ${\lambda}$형(型)의 N-말단(末端) 및 그 주변(周邊)의 아미노산배열(酸配列)을 결정(決定)하기 위하여 본(本) 실험(實輸)이 시도(試圖)되었던바 그 결과(結果)는 다음과 같다. 1) Bence Jones 단백질(蛋白質)을 Pronase와 Chymotrypsin으로 분해(分解)하여 얻은 Peptide중에서 Im Pr-M 및 Im Ch-M와 Ik Ch-M을 Dowex $50{\times}2$ column $1{\times}20$cm)와 Dowex $1{\times}2$ column $(0.9{\times}50{\;}cm)$을 사용(使用)하여 분리(分離)하였다. 2) ${\lambda}$형(型) Bence Jones단백질(蛋白質)의 N-말단(末端)은pyrroglutamic acid로 되어 있음을 alkali반응(反應)과 고압여지전기영동법(高壓濾紙電氣泳動法)으로 확인(確認)하였다. 3) 농염산(濃鹽酸)(12N) 반응(反應)($27^{\circ}C$, 15시간(時間))을 이용(利用)하며 Peptide중(中)의 Serine부(部)를 선택적(選擇的)으로 절단(切斷)할 수 있었다. 4) 이들 Peptide의 아미노배열순서(配列順序)는 Edman의 PTC법(法)과 소거법(消去法) 및 CarboBypeptidase A를 사용(使用)하여 결정(決定)하였다. 5) 분리(分離)한 Peptide의 아미노산배열순서(酸配列順序)는 다음과 같았다. $Im\;Ch-M\;PCA{\cdot}Ser{\cdot}Val{\cdot}Leu$ $Ik\;Ch-M\;PCA{\cdot}Ser{\cdot}Ala{\cdot}Leu1$