• Title/Summary/Keyword: Reversed phase $C_{18}$ column

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Determination of Costunolide from Aucklandiae Radix and Substitutive Herbs by Reversed-Phase HPLC (HPLC를 이용한 목향 및 유사 한약재에 함유된 Costunolide 비교)

  • Oh, Ju-Hee;Hong, Seon-Pyo;Choi, Ho-Young;Park, Yong-Ki;Lee, Je-Hyun
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.61-66
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    • 2008
  • Objecives : Aucklandiae Radix is a root of Aucklandia lappa which has been widely used for regulating the flow of vital energy, invigorating the spleen, alleviating pain. Aucklandiae Radix contains the costunolide which is the main ingredient. The substitutive Aucklandiae Radix are Inulae helenii Radix, Aristolchiae Radix, Vladimiriae Radix, and Inulae racemosi Radix in Korea and China. This paper is analysised and compared the costunolide and HPLC pattern in Aucklandiae Radix and substitute herbs. Methods : Chromatographic separation performed using C18 column(Luna 5 u, 250 mm ${\times}$ 4.6 mm) with a mixture of methanol and water(65:35)(v/v). The analyses detected at UV(210 nm). Results : Optimal extraction condition of costunolide was 100% methanol for 2hr. Costunolide was detected in Aucklandiae Radix and Vladimiriae Radix, but other herbs were not detected. In Korea herbal market, Aristolchiae Radix merchandise was identified as the imported Inulae helenii Radix. Conclusions : According to above results, this method was useful identified to Aucklandiae Radix and substitutive herbs. In Korea herbal market, Aristolchiae Radix was identified as Inulae helenii Radix.

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Determination of Recombinant Human Epidermal Growth factor (rhEGF) in a Pharmaceutical Formulation by High Performance Liquid Chromatography with Electrochemical Detection

  • Lee, Kang-Woo;Hwang, Kyung-Hwa;Kim, Chang-Soo;Han, Kun;Chung, Youn-Bok;Park, Jeong-Sook;Lee, Yong-Moon;Moon, Dong-Cheul
    • Archives of Pharmacal Research
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    • v.24 no.4
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    • pp.355-359
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    • 2001
  • A novel HPLC method with electrochemical detection has been developed for the determination of recombinant human epidermal growth factor (rhEGF) in pharmaceutical products. rhEGF was separated from other components in formulation on a reversed-phase C18 column with 24% acetonitrile in 0.1 M phosphate buffer (pH 4.75). The optimum electrochemical oxidation of EGF was obtained at 0.85 V vs. Ag/AgCl in a glassy carbon working electrode due to electroactive tyrosine, tryptophan, methionine, and arginine residues. The quantitation range was from 1.0 to 200 ng of rhEGF with the linear correlation coefficient greater than 0.999. The method was successfully applied for the quantitation of rhEGF in a pharmaceutical preparation.

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Evaluation of the functional phenolic compounds as to various ripening phases of the reddish rice cultivar(Oryza sativa, Superhongmi)

  • Ham, Tae-Ho;Kwon, Soon Wook;Kwon, Young In;Ryu, Su-Noh
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.272-272
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    • 2017
  • Recently, consumer's demands are increasing for new natural functional and healthful food. These demands have encouraged a better understanding of the biochemical, chemical and nutritional composition of plant products. Superhongmi rice variety was derived from a cross between CG2-3-5-2-6-2 (Heugjinju/Suwon 425) having reddish brown color and Daeribbyeo 1 having a large grain size. Superhongmi rice headed on Sep. 5th and has 94.7 cm culm length. Taxifolin, a phenolic compound in superhongmi rice extract had high ${\alpha}-glucosidase$ inhibitory activity. The ${\alpha}-glucosidase$ inhibitory activity of the superhongmi rice extracts correlated to the taxifolin content and antioxidant activity of the extracts. A new method for quantitative determination of taxifolin in superhongmi rice variety by high performance liquid chromatography was established. A reversed-phase system with Tosoh TSK-gel $C_{18}$ column using 60% methanol in water(pH 2.4) as a mobile phase was developed. Taxifolin was detected at 280 nm and the analysis was successfully carried out within 40 min. In ripening phase, the amount of taxifolin showed a mild decreasing slope. The highest contents was 59.27mg in 100g seed on the 30 days after heading(DAH). The optimum harvesting time, considering taxifolin content, maturity rate and thousand seed weight(TSW) was DAH 50. These results suggest that superhongmi rice which has high taxifolin content has the potential to contribute as a dietary supplement for controlling hyperglycemia and oxidative stress-linked diabetes complications.

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Changes on the Quercitrin Content in the Preparation for the Leaves of Cedrela sinensis (참죽나무잎의 조리시 quercitrin의 함량 변화)

  • 박종철;전순실;김성환
    • Korean journal of food and cookery science
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    • v.11 no.3
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    • pp.303-308
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    • 1995
  • When we prepared the cooked namul, bugak and kimchi by the leaves of Cedrela sinensis, the changes of quercitrin which was isolated from the leaves of this plant in each preparations were analysed by HPLC. Separation by reversed phase chromatography on u-Bondapak C/Sub 18/ column was achieved by isocratic elution with THF-dioxane-MeOH-HOAc-5% H/Sub 3/PO/Sub 4/-H/Sub 2/O (145: 125:50:20:2:658). When Kimchi was stored at 5$^{\circ}C$ for 20 days, the content of quercitrin in the methanol extract of Kimchi was 7.21% and Kimchi at 20$^{\circ}C$ was reduced by 5.78% (w/w). Contents of quercitrin in the leaves of Cedrela sinensis kimchi obsered to be gradually decreased during storage at 5$^{\circ}C$ and 20$^{\circ}C$. Contents of quercitrin of stored kimchi at 5$^{\circ}C$ was higher than that stored at 20$^{\circ}C$. The contents of quercitrin in Namul and Bugak were 13.06 and 5.03% (w/w), respectively, which were lower than control.

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Study of the Separation and Elution Behavior of Phenols as Priority Pollutants in Reversed Phase Liquid Chromatography (역상 액체 크로마토그래피에서 유기오염물질로서의 페놀류들의 분리 및 용리거동에 관한 연구)

  • Dai Woon Lee;Sun Kyung Lee;Keun Sung Yook;Won Lee
    • Journal of the Korean Chemical Society
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    • v.33 no.3
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    • pp.287-294
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    • 1989
  • The optimum condition for the separation of priority pollutant phenols using isocratic elution has been determined. The elution behavior of eleven phenols has been also studied to interpret the retention. The reversed phase liquid chromatographic methods were performed on a ${\mu}$-Bondapak $C_{18}$ column with methanol-water, acetonitrile-water, and THF water mixtures as mobile phases. The COF method, where Snyder's solvent triangle concept was combined with a mixture-design statistical technique, was used to optimize the strength and selectivity of solvents for the separation of phenols. The optimum solvent composition, which gives a complete separation of eleven phenols, was found to be $MeOH:ACN:H_2O$ = 7:40:53. The plots of ln k' vs. -${\Delta}H^{\circ}$ and ${\Sigma}{\pi}$ of phenols showed relatively good linearities. Effect of van der Waals volume, pi-energy and hydrogen bonding on the retention of phenols were investigated. The following equation with the correlation coefficient of 0.9927 for ACN-water solvent system was obtained; $log^{k'}=2.515{\times}10^{-2}VWV-1.301{\times}10^{-1}E-3.674{\times}10^{-1}$

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Sensitive Determination of Pinaverium Bromide in Human Plasma by LC-ESI-MS/MS : Applicability to Oral Bioavailability Determination (LC-ESI-MS/MS를 이용한 생체시료 중 브롬화피나베리움의 고감도 분석 및 이를 이용한 생체이용률 평가)

  • Park, Seok;Lee, Ye-Rie;Kim, Ho-Hyun;Lee, Hee-Joo;Kim, Yoon-Gyoon;Youm, Jeong-Rok;Han, Sang-Beom
    • Journal of Pharmaceutical Investigation
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    • v.34 no.6
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    • pp.513-519
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    • 2004
  • A sensitive method for quantification of pinaverium bromide in human plasma was established using liquid chromatography-electrospray ionization tandem mass spectrometry(LC-ESI-MS/MS). Glimepiride was used as internal standard. Pinaverium bromide and internal standard in plasma sample were extracted using tert-butylmethylether(TBME). A centrifuged upper layer was then evaporated and reconstituted with mobile phase of acetonitrile-5 mM ammonium formate (80/20, pH 3.0). The reconstituted samples were injected into a $C_{18}$ reversed-phase column. Using MS/MS with multiple reaction monitoring (MRM) mode, pinaverium and glimepirde were detected without severe interference from human plasma matrix. Pinaverium produced a protonated precursor ion $([M+H]^+)$ at m/z 510.3 and a corresponding product ion at m/z 228.9. Internal standard produced a protonated precursor ion $([M+H]^+)$ at m/z 491.5 and a corresponding product ion at m/z 352.0. Detection of pinaverium bromide in human plasma was accurate and precise, with limit of quantitation at 0.5 ng/ml. The method has been successfully applied to bioavailability study of pinaverium bromide tablet in Korean healthy male volunteers. Pharmacokinetic parameters such as $AUC_t,\;C_{max},\;T_{max},\;K_{el}\;and\;t_{1/2}$ were calculated.

Study on analytical method of fluoroquinolone residues in eggs by LC/MS/MS (LC/MS/MS를 이용한 식용란 중 fluoroquinolone계 항균물질의 분석법에 관한 연구)

  • Choi, You-Jeong;Yun, I-Ran;Nam, Sang-Yun;Park, Young-Ho;Kim, Byeong-Hun;Son, Seong-Gi
    • Korean Journal of Veterinary Service
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    • v.30 no.1
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    • pp.13-21
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    • 2007
  • An atmospheric pressure chemical ionization (APcI) LC/MS/MS method was developed for the simultaneous analysis of fluoroquinolones (norfloxacin, ciprofloxacin, enrofloxacin, danofloxacin) residues in eggs. The spiked and blank samples were extracted from whole eggs using 50mM phosphate buffer (pH 7.4). The extract was cleaned up by passage though $Oasis^{(R)}$ MAX extraction cartridge for solid-phase extraction followed by elution with 4% formic acid in methanol. The extract of sample was separated on a Waters $Atlantis^{TM}$ $dC_{18}$ reversed-phase column ($4.6{\times}150mm,\;5{\mu}m$) and analyzed by APcI positive mode mass spectrometry. The mobile phase consists of aqueous 0.2% nonafluoropentanoic acid (NFPA) and methanol. Multiple reaction monitoring (MRM) using the precursor to product ion combinations of m/z $320\;{\dashrightarrow}\;302,\;332\;{\dashrightarrow}\;314,\;360\;{\dashrightarrow}\;342$ and m/z $358\;{\dashrightarrow}\;340$ were used to quantify norfloxacin (NOR), ciprofloxacin (CIP), enrofloxacin (ENR) and danofloxacin (DAN), respectively. The limits of quantification (LOQ) were 7.8ppb for NOR, 8.5ppb for CIP, 8.9ppb for ENR, and 4.8ppb for DAN. Average recoveries of fortified sample at levels of 0.025 to 0.1 ppm were estimated 71.29% for NOR, 75.27% for CIP, 85.51% for ENR and 81.22% for DAN. These results could be applied for the confirmation and quantification in eggs.

Separation and Determination of Acteoside in Pedicularis resupinata var. oppositifolia by Ion Pair Liquid Chromatography (이온쌍 액체 크로마토그래피에 의한 마주송이풀 중의 Acteoside의 분리와 정량)

  • Yun, Young Ja;Yu, Gu Yong
    • Analytical Science and Technology
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    • v.8 no.2
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    • pp.161-166
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    • 1995
  • Determination of acteoside in Pedicularis resupinata var. oppositifolia has been studied using ion pair liquid chromatography. Sample was extracted with 40mL methanol for 4 hrs. The extract was cleaned up by using Sep-Pak $C_{18}$ catridge and 8mL aqueous methanol eluent(methanol 50%, water 50%, phosphate buffer pH=8.0). Its determination was performed by means of IP-HPLC with a Hamilton PRP-1 polystyrene-divinylbenzene reversed phase column($15cm{\times}4.6mm$ i. d., $5{\mu}m$) and an aqueous methanol eluent(methanol 60%, water 40% phosphate buffer pH=8.2) containing of $5.0{\times}10^{-3}M$ tetrabutylammonium bromide. The established method was applied to the sample that was collected in area of Pyung Chang gun. As a result, its content ranges showed to be 0.062~0.076%.

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Determination of Aflatoxins Using High-Performance Liquid Chromatography and Fluorescence or UV Absorbence Detection (HPLC에 의한 aflatoxin 분석법에 관한 연구 형광 및 자외선 흡광 검출의 비교)

  • 김종규;강회양;민경진
    • Journal of Environmental Health Sciences
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    • v.22 no.1
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    • pp.36-44
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    • 1996
  • A comparison was made of two detection methods(UV absorbence detection and fluorescence detection with pre-column derivatization, with trifluoroacetic acid) coupled with HPLC for the simultaneous determination of aflatoxin $B_1, B_2, G_1$ and $G_2$. A good separation of the four aflatoxins was achieved on a reversed-phase $C_{18}$ column (30 cm x 3.9 mm) with methanol-acetonitrile-water(20+20+60) for absorbence detection or acetonitrile-water(25+75) for fluorescence detection at the flow rate of 1.0 ml/min. The calibration graphs were linear over the ranges 100 ppb-1 ppm for $B_1/G_1$ and 30~300 ppb for $B_2/G_1$ with absorbence detection, and 1~500 ppb for $B_1/G_1$ and 0.3~150 ppb for $B_2/G_2$ with fluorescence detection. The correlation coefficients were greater than 0.94 and 0.99 for absorbance detection and for fluorescence detection, respectively. The detection limit was 100 ng for $B_1/G_1$ and 30 ng for $B_2/G_2$ with absorbence detection, and 1 ng for $B_1/G_1$ and 0.3 ng for $B_2/G_2$ with fluorescence detection. Recovery rates of aflatoxin $B_1, B_2, G_1$ and $G_2$ added to yeast-extract sucrose broth medium were 66.6%, 59.4%, 67.5% and 59.2%, respectively, for absorbence detection and 82.9%, 71.5%, 80.0% and 69.3%, respectively, for fluorescence detection. The four aflatoxins in culture medium were quantitatively detected by the two methods. The aflatoxins in the rice sample were not detected the absorbence detection method, but were below 10 ppb using the fluorescence detection method. Analysis of aflatoxins by both the absorbence and fluorescence methods coupled with HPLC showed acceptable linearity and good recovery. The absorbence detection was less timeconsuming and safer for treatment. The fluorescence detection was more elective and sensitive though elevated $B_1$ and $G_1$ contents were determined from the TFA-induced conversion of $B_1$ to $B_{2a}$ and $G_1$ to $G_{2a}$.

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Quantitative Determination of Diterpenoids from the Roots of Aralia cordata

  • Lee, Mi-Kyoung;Hung, Tran Manh;Min, Byung-Sun;Lee, Ik-Soo;Na, Min-Kyun;Woo, Mi-Hee;Son, Jong-Keun;Kim, Young-Ho;Choi, Jae-Sue;Bae, Ki-Hwan
    • Natural Product Sciences
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    • v.15 no.1
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    • pp.50-54
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    • 2009
  • A simple and reliable reversed phase HPLC method was developed to determine pharmacologically active compounds ent-continentalic acid, kaurenoic acid, and continentalic acid from the roots of Aralia cordata. Quantitative analysis of diterpenoids of A. cordata were performed by reverse phase C-18 column using a isocratic of acetonitrile : 0.1% trifluoroacetic acid(70:30) with UV detection at 205 nm. The correlation coefficients of each calibration curve of ent-continentalic acid, kaurenoic acid, and continentalic acid were 0.9987, 0.9983, and 0.9986, respectively. The detection limit of ent-continentalic acid, kaurenoic acid, and continentalic acid were $0.5{\sim}1.0\;{\mu}g/mL$, respectively. The contents of ent-continentalic acid (1), kaurenoic acid (2), and continentalic acid (3) of the roots of A. cordata collected from seventeen district markets in Korea were $0.09{\sim}0.75\;mg/g$, $1.09{\sim}5.43\;mg/g$, and $2.69{\sim}9.08\;mg/g$, respectively.