• Title/Summary/Keyword: reversed phase-HPLC

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Simultaneous Analysis of Both Lactone Form and Acid Form Monacolin K in Red Yeast Rice by RP-HPLC (역상 HPLC에 의한 홍국 중의 락톤 및 산성 모나콜린 K의 동시분석법)

  • Moon, Young-ja;Wang, Qi-jun;Xu, Bao-jun;Li, Chang-tian;Kim, Jae-hoon;Mo, En-kyeng;Baek, Seoung-young;Il Kwon;Sung, Chang-keun
    • The Korean Journal of Food And Nutrition
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    • v.14 no.6
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    • pp.521-526
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    • 2001
  • A method for the simultaneous and precise determination of lactone form and acid form monacolin K in red yeast rice by HPLC was developed in this study. The standard of acid form monacolin K was prepared by alkaline hydrolysis of its lactone form, which was purchased from Sigma company. The optimum HPLC system for the separation and quantification of acid form and lactone form monacolin K is based on the reversed-phase column, and the acidified mobile phase consisting of acetonitrile : 0.1% trifluoroacetic acid (TFA) water soln : 62 :38, the low limit detection amount was 5 ng (i.e.10 $\mu$l injection of 0.5 $\mu\textrm{g}$/ml) . And the optimal extracting system for monacolins in red rice was also presented here.

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Separation and Elution Behaviors of Some Metal-2-hydroxyarylazopyrazolone Chelates by Reversed Phase High Performance Liquid Chromatography (I) (역상 액체 크로마토그래피에 의한 금속-2-hydroxyarylazopyrazolone 유도체 킬레이트의 용리거동 및 동시분리에 관한 연구 (I))

  • Lee, Chang-Heon;Kang, Chang-Hee;Kim, Eun-Kyung;Lee, Won
    • Analytical Science and Technology
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    • v.7 no.1
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    • pp.103-114
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    • 1994
  • The elution behaviors of Ni(II), Cu(II), Co(II), and Cr(II) in 1-phenyl-3-methyl-4(2-hydroxy-5-X-phenylazo)-5-pyrazolone, [Pm(2-OH)(5-X)PaPz](X=H, $CH_3$, $NO_2$, Cl) chelates have been studied by reversed phase HPLC. Thirteen metal-[Pm(2-OH)(5-X)PaPz] chelates were prepared and characterized by UV, IR, MS, and ICP spectroscopic methods. These metal-2-hydroxyarylazopyrazolone chelates were successfully separated on Novapak-$C_{18}$ column using methanol/water mixtures as a mobile phases. It was found that the chelates were eluted properly in an acceptable range of the capacity factor value($0{\leq}log\;k^{\prime}{\leq}1$). The dependence of the capacity factor(log k') on the volume fraction of water in the binary mobile phase showed a good linearity. Also, there was a good linear dependence of the capacity factor on the liquid-liquid extraction distribution ratio($D_c$) in methanol-water/n-pentadecane extraction system by the batch method. It suggested that the retention of the chelates in the reverse phase liquid chromatographic system be largely due to the solvophobic effect.

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Determination of Chloramphenicol in Milk by High Performance Liquid Chromatography (HPLC를 이용한 우유중의 클로람페니콜의 정량 분석)

  • 김경례;김정한;최경숙
    • YAKHAK HOEJI
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    • v.29 no.1
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    • pp.50-54
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    • 1985
  • Seven different sorbents were evaluated for their adsorptivity and desorptivity of antibiotic, chloramphenicol. Among the sorbents studied, Carbopak B was found to be the most efficient in enriching the chloramphenicol from dilute aqueous solution. Interfering components in the milk matrix could be washed off by water and petroleum ether from Carbopak B column, while the chloramphenicol was retained on the surface of Carbopak B. The method of simple and efficient purification and enrichment of chloramphenicol using Carbopak B, followed by quantitative analysis employing $C_{18}$ reversed phase high performance liquid chromatography has been applied to the determination of chloramphenicol in milk.

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Resolution of the Triacylglycerols Containing Conjugate Trienoic Acids into Their Molecular Species by HPLC in the Reversed-phase and Silver Ion Mode (Reversed-phase 및 $Ag^{+}$-HPLC에 의한 Conjugate Trienoic Acid 함유(含有) Triacylglycerol 분자종(分子種)의 상호분리(相互分離))

  • Kim, Seong-Jin;Woo, Hyo-Kyeng;Joh, Yong-Goe
    • Journal of the Korean Applied Science and Technology
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    • v.18 no.3
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    • pp.197-213
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    • 2001
  • Conjugate trienoic acids (CTA) occurred in triacylglycerols (TGs) of the seed oils of Trichosanthes kirilowii, Momordica charantia and Aleurites fordii, and they were easily converted to their methyl esters in a mixture of sodium methoxide-methanol without any structural destruction. The main fatty acids in triacylglycerol (TG) fraction of the seed oils of Trichosanthes kirilowii are $C_{18:2{\omega}6}$ (32.2 mol %), $C_{18:3{\;}9c.11t,13c}$ (38.0 mol %) and $C_{18:1{\omega}9}$ (11.8 mol %), followed with $C_{16:0}$ (4.8 mol %) and $C_{18:0}$ (3.1 mol %). The TG fraction was resolved into 20 TG molecular species according to the partition number (PN) by reversed-phase (RP)-HPLC. The main TG species were $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$, of which amounts reached 63 mol % of total TG molecular species. The TG sample was fractionated into 11 fractions according to the number of double bond in the molecule by $Ag^{+}-HPLC$ and the species of $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$ were also eluted as main components. The TG species containing CTA showed unusual behaviours in the order of elution by HPLC ; first, TG moleular species of $DT_{c2}$ (D; dienoic acid, $T_{c}$; punicic acid, $T_{ci}$; ${\alpha}-eleostearic$ acid, M ; monoenoic acid, $S_{t}$; stearic acid) was eluted earlier than $Mt_{c2}$, although they have the same PN number of 40, and, secondly, the species of $DT_{ci2}$ with eight double bonds was eluted earlier than that of $D_2T_{ci}$ with seven double bonds. Intact TG of the seed oils of Momordica charantia contained mainly fatty acids such as $C_{18:3{\omega}9c,11t,13t}$ (57.7 mol %), $C_{18:1{\omega}9}$ (17.4 mol %), $C_{18:0}$ (12.3 mol %) and $C_{18:2{\omega}6}$ (10.6 mol %), and was classified into 13 fractions by RP-HPLC. The main TG species were as follows ; $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.1 mol %] and $S_{t}T_{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_2$, 33.9 mol %] comprising about 73 mol % of total TG species, accompanied by $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 7.3 mol %], $D_{2}T_{ci}$ [$ (C_{18:2{\omega}6})_{2}(C_{18:3\;9c,11t,13t})$, 3.6 mol %] and $MDT_{ci}$ [$(C_{18:1{\omega}9})(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})$, 3.5 mol %]. Simple TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_3]$ was present in a small amount of 1.4 mol %, but other simple TG species were not detected. The TG was also resolved into 11 fractions according to the number of double bond by $Ag^{+}-HPLC$, and the species were mainly occupied by $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.4 mol %] and $S_tT-{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_{2}$, 35.4 mol %] $DT_{ci2}$ species with eight double bonds was also developed faster than $D_2T_{ci}$ one with seven double bonds as indicated in the analysis of TG of the seed oils of T. kirilowii, and $MT_{ci2}$ species with cis, trans, trans-configurated double bond was eluted earlier than $MT_{c2}$ species with cis, trans, cis-configurated double bond. The main components of fatty acid in total TG fraction isolated from the seed oils of of Aleurites fordii were in the following order ; $C_{18:3\;9c,11t,13t}$ (81.2 mol %)> $C_{18:2{\omega}6}$ (8.5 mol %)> $C_{18:1{\omega}9}$ (5.4 mol %)$. With resolution of the TG by RP-HPLC, eight fractions such as $T_{ci3}$, $Dt_{ci2}$, $D_{2}T_{ci}$, $MT_{ci2}$, $PT_{ci2}$ (P; palmitic acid), $PMT_{ci}$, $PDT_{ci}$ and $S_{t}T_{ci2}$ ($S_{t}$; stearic acid) were isolated, respectively. TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_{3}$, 54.2 mol %], $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 15.0 mol %] and $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3 9c,11t,13t})_{2}$, 14.8 mol %] were present as main species.

HPLC Separation of Isoquinoline Alkaloids for Quality Control of Corydalis species

  • Kim, Eun-Kyung;Jeong, Eun-Kyung;Han, Sang-Beom;Jung, Jee-H.;Hong, Jong-Ki
    • Bulletin of the Korean Chemical Society
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    • v.32 no.10
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    • pp.3597-3602
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    • 2011
  • A simple and rapid analytical method was developed for the determination of eight isoquinoline alkaloids in Corydalis species. Eight isoquinoline alkaloids, including 2 aporphine alkaloids (isocorydine and glaucine) and 6 protoberberine alkaloids (coptisine, palmatine, berberine, canadine, corydaline, and tetrahydrocoptisine) were used as chemical markers which have a various biological activity and determined for quality control of Corydalis (C.) species (C. ternata, C. yanhusuo, and C. decumbens). To evaluate the quality of these herbal medicines, LC chromatographic separation of alkaloids were preferentially investigated on reversed-phase C18 column with pH variation and composition of mobile phase. In addition, the separation of these alkaloids in herbal extracts was found to be significantly affected on mobile phase composition using gradient elution. Especially for C. yanhusuo extract, berberine was seriously interfered with other alkaloid extracted from sample matrix when mobile phase composition was not optimized. As results, these compounds were successfully separated within 28 min using 10 mM ammonium acetate containing 0.2% triethylamine (adjusted at pH 5.0) as a mobile phase with gradient elution. On the basis of optimized HPLC conditions, 23 different Corydalis species samples were analyzed for the determination of alkaloid levels. In addition, principal component analysis (PCA) combined with the chromatographic data could be successfully classified the different geographic origin samples.

Supercritical Fluid Extraction of Catecnin Compounds from Green Tea (녹차에서 카테킨 화합물의 초임계 유제 추출)

  • 나영진;이윤우;김재덕;노경호
    • KSBB Journal
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    • v.16 no.4
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    • pp.327-331
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    • 2001
  • Catechin compounds from the Green tea cultivated in Bosung (Chollanamdo) were extracted using supercritical fluid and various additives, and analyzed by reversed-phase high performance liquid chromatography (RP-HPLC). The chromatographic column was packed with LiChrospher 100RP-18(15 $\mu$m), and water was used as the mobile phase with 0.05% phosphoric acid and acetonitrile. Gradient election was applied to separate EGCG by changing the mobile phase compositions. Comparing the extraction yield of three different types of supercritical fluids, pure CO$_2$, with additives of water and ethanol (5 wt,%), the extraction amount of EGCG was most abundant in the suprcritical CO$_2$with ethanol. However, more was extracted and pure higher purity was achieved by solvent extraction using ethanol.

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Impurity Profiling and Quantification of Sudan III Dyes by HPLC-selective UV Detection

  • Yang, Ki Ryeol;Hong, Ji Yeon;Yoon, Soo Hwan;Hong, Jongki
    • Bulletin of the Korean Chemical Society
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    • v.35 no.3
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    • pp.765-769
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    • 2014
  • An analytical methodology was developed for qualitative and quantitative impurity profiling of the coloring agent Sudan III by high-performance liquid chromatography (HPLC)-diode array detection (DAD). The impurities in commercial Sudan III were characterized by comparison of their retention times and UV spectra with those of authentic standards. Four impurities regulated by International Committees in Sudan III were quantified by HPLC-selective UV detection. The impurities in Sudan dye were successfully separated on a reversed phase C18-column within 25 min and sensitively detected by UV spectrometry at two selective wavelengths. Method validation was conducted to determine linearity, precision, accuracy, and limit of quantification (LOQ). The linear dynamic range extended from 0.002 to 4.0%, with a correlation coefficient (R2) greater than 0.995. The LOQs of the impurities ranged from 8.04 to $54.29{\mu}g/mg$. Based on the established method, the levels of regulated impurities in five commercial Sudan III dyes were determined.

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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Identification of IY81149 and Its Metabolites in the Rat Plasma Using the On-Line HPLC/ESI Mass Spectrometry

  • Myung, Seung-Woon;Min, Hye-Ki;Jin, Chang-Bae;Kim, Myung-Soo;Lee, Seung-Mok;Chung, Gi-Ju;Park, Seong-Jun;Kim, Dong-Yeon;Cho, Hyun-Woo
    • Archives of Pharmacal Research
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    • v.22 no.2
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    • pp.189-193
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    • 1999
  • Reversed-phase high-performance liquid chromatography/mass spectrometry (HPLC/MS) with an eletcrospray ionization (ESI) interface was applied to the identification of metabolites of IY81149 in the rat plasma. Fragments obtained using collision-induced dissociation (CID) in both positive and negative modes were utilized to elucidate the structure of metabolites. The eluent from the conventional HPLC column was split and directly introduced into an ESI-mass spectrometer for the identification of the structures. the CID technique allowed the sensitive identification of sulfonyl-IY81149 and hydroxy-IY81149 from the rat plasma.

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