• Title/Summary/Keyword: HPLC Separation

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Separation and Determination of Saikosaponins in Bupleuri Radix with HPLC (HPLC에 의한 시호(柴胡) Saponin의 분리 및 정량)

  • Han, Dae-Suk;Lee, Dug-Keun
    • Korean Journal of Pharmacognosy
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    • v.16 no.3
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    • pp.175-179
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    • 1985
  • The optimal condition for the determination of saikosaponin a and d, the major pharmacologically active saponins of the roots of Bupleurum falcatum, was studied with the conversion of these saponins into diene saponins $(saikosaponin\;b_1\;and\;b_2)$. The complete separation and quantitative analysis of these saponins were performed by the method of high performance liquid chromatography using $NH_2$ column. The conversion of saikosaponin a and d into diene saponins under gastric pH was calculated. Thirty-three percent of saikosaponin a was converted to saikosaponin $b_1$ and 63 percent of saikosaponin d was converted to saikosaponin $b_2$.

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Determination of Terbutaline Enantiomers in Human Plasma by Coupled Achiral-Chiral High Performance Liquid Chromatography

  • Kim, Kyeong-Ho;Kim, Hyun-Ju;Hong, Seon-Pyo;Shin, Sang-Deok
    • Archives of Pharmacal Research
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    • v.23 no.5
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    • pp.441-445
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    • 2000
  • Achiral-chiral column switching HPLC assay was developed to allow the separation and quantification of the enantiomers of terbutaline in human plasma by means of fluorescence detection. Plasma samples were prepared by solid-phase extraction with sep-pak silica, followed by HPLC assay. The enantiomers of terbutaline and the internal standard were separated from the biological matrix on a silica column, and the two enantiomers were resolved and quantified on a Sumichiral OA-4900 column. The two columns were connected by a switching valve equipped with silica trap column, The trap column was used to concentrate the terbutaline in the eluent from the achiral column before back flushing onto the chiral phase. For each enantiomers, the assay was linear between 2.5-125 ng/$m\ell$ (r=0.9999) and detection limit was 1.0 ng/$m\ell$ .

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Investigation of Enantiomer Separation Using Chiral Crown Ethers as Chiral Selectors

  • Lee, Wonjae
    • Journal of Integrative Natural Science
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    • v.9 no.1
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    • pp.28-34
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    • 2016
  • A number of chiral selectors have been developed and applied for enantiomer separation of a variety of chiral compounds. Among these chiral selectors are chiral crown ethers, a class of synthetic host polyether molecules that bind protonated chiral primary amines with high selectivity and affinity. In this paper, two important chiral crown ethers as chiral selectors of bis-(1,1'-binaphthyl)-22-crown-6 and (18-crown-6)-2,3,11,12-tetracarboxylic acid (18-C-6-TA) are focused. They have been widely used to resolve the enantiomers of chiral compounds containing a primary amino moiety using chiral stationary phases (CSPs) or chiral selectors by high-performance liquid chromatography (HPLC), capillary electrophoresis (CE) and so on in chirotechnology. Also, it was described that the commercially available covalent type HPLC CSPs derived from (+)- and (-)-18-C-6-TA have been developed and successfully applied for the resolution of various primary amino compounds including amino acids.

HPLC Method for the Determination of Nicorandil in Human Plasma

  • Park, Sun-Hee;Shin, In-Chul
    • Biomolecules & Therapeutics
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    • v.16 no.2
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    • pp.168-172
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    • 2008
  • The present study is to determine of sensitive nicorandil analysis method using HPLC and measure the pharmacokinetics parameters (bioavailability, $C_{max}$, $T_{max}$, Ke, $T_{1/2}$) of nicorandil (5 mg, Tab; Choongwae Pharma Corporation). Plasma (500 ul) was mixed with furosemide (internal standard, 500 ug/ml). Detection wavelength was 256 nm. The mixture of 0.01 M ammonium acetate and acetonitrile 80:20 (v/v) was used mobile phase. The HPLC separation was accomplished on ODC reverse HPLC column. The nicorandil was analyzed by a HPLC system, which consists of CAPCELL PAK C18 column (5 ${\mu}$m, 4.6 × 150 mm) and a chromatography data analysis S/W, using a isocratic mobile phase (mixture of 0.01 M ammonium acetate and acetonitrile 80:20 ) at 1.0 ml/min. Its sensitivity, selectivity, accuracy and precision must be adequate for the bioavailabilty study of nicorandil, and the linearity ($r^2$ ≥ 0.9994) of nicorandil was also proved in the range of 0.05 ug/ml . 3 ug/ml. The pharmacokinetic parameters of nicorandil (5 mg) tablets were measured as the follow. AUC: 0.19 ug/ml·hr, $C_{max}$: 0.14 ug/ml, $t_{max}$: 0.58 hr, Ke: 0.11 hr., $t_{1/2\beta}$: 6.76 hrs. This method is simple and sensitive HPLC method using UV detector for determination of nicorandil in human plasma.

Quantitative Analysis of Bioactive Marker Compounds from Cinnamomi Ramulus and Cinnamomi Cortex by HPLC-UV

  • Jeong, Su Yang;Zhao, Bing Tian;Moon, Dong Cheul;Kang, Jong Seong;Lee, Je Hyun;Min, Byung Sun;Son, Jong Keun;Woo, Mi Hee
    • Natural Product Sciences
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    • v.19 no.1
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    • pp.28-35
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    • 2013
  • In this study, quantitative and pattern recognition analysis for the quality evaluation of Cinnamomi Ramulus and Cinnamomi Cortex using HPLC/UV was developed. For quantitative analysis, three major bioactive compounds were determined. The separation conditions employed for HPLC/UV were optimized using an ODS $C_{18}$ column ($250{\times}4.6$ mm, 5 ${\mu}m$) with gradient conditions of acetonitrile and water as the mobile phase, at a flow rate of 1.0 mL/min and a detection wavelength of 265 nm. This method was fully validated with respect to linearity, accuracy, precision, recovery, and robustness. The HPLC/UV method was applied successfully to the quantification of three major compounds in the extract of Cinnamomi Ramulus and Cinnamomi Cortex. The HPLC analytical method for pattern recognition analysis was validated by repeated analysis of thirty eight Cinnamomi Ramulus and thirty five Cinnamomi Cortex samples. The results indicate that the established HPLC/UV method is suitable for quantitative analysis.

Quantitative and Pattern Recognition Analyses for the Quality Evaluation of Magnoliae Flos by HPLC

  • Fang, Zhe;Shen, Chang Min;Moon, Dong-Cheul;Son, Kun-Ho;Son, Jong-Keun;Woo, Mi-Hee
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3371-3381
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    • 2010
  • In this study, quantitative and pattern recognition analysis for the quality evaluation of Magnoliae Flos using HPLC/UV was developed. For quantitative analysis, eleven major bioactive lignan compounds were determined. The separation conditions employed for HPLC/UV were optimized using ODS $C_{18}$ column ($250{\times}4.6\;mm$, $5\;{\mu}m$) with isocratic elution of acetonitrile and water with 1% acetic acid as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 278 nm. These methods were fully validated with respect to the linearity, accuracy, precision, recovery, and robustness. The HPLC/UV method was applied successfully to the quantification of eleven major compounds in the extract of Magnoliae Flos. The HPLC analytical method for pattern recognition analysis was validated by repeated analysis of twenty one reference samples corresponding to seven different species of Magnoliae Flos and nine samples purchased from market. The results indicate that the established HPLC/UV method is suitable for the quantitative analysis and quality control of multi-components in Magnoliae Flos.

Quantitative and Pattern Recognition Analyses for the Quality Evaluation of Cimicifugae Rhizoma by HPLC

  • Fang, Zhe;Moon, Dong-Cheul;Son, Kun-Ho;Son, Jong-Keun;Min, Byung-Sun;Woo, Mi-Hee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.1
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    • pp.239-246
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    • 2011
  • In this study, quantitative and pattern recognition analysis for the quality evaluation of Cimicifugae Rhizoma using HPLC/UV was developed. For quantitative analysis, three major bioactive phenolic compounds were determined. The separation conditions employed for HPLC/UV were optimized using ODS $C_{18}$ column ($250{\times}4.6mm$, $5{\mu}M$) with isocratic elution of acetonitrile and water with 0.1% phosphoric acid as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 323 nm. These methods were fully validated with respect to the linearity, accuracy, precision, recovery, and robustness. The HPLC/UV method was applied successfully to the quantification of three major compounds in the extract of Cimicifugae Rhizoma. The HPLC analytical method for pattern recognition analysis was validated by repeated analysis of twelve reference samples corresponding to five different species of Cimicifugae Rhizoma and seventeen samples purchased from markets. The results indicate that the established HPLC/UV method is suitable for the quantitative analysis and quality control of multi-components in Cimicifugae Rhizoma.

Enantiomeric Purity Test of Bevantolol by Reversed-Phase High Performance Liquid Chromatography after Derivatization with 2,3,4,6-tetra-O-acetyl-$\beta$-D-glucopyranosyl Isothiocyanate

  • Kim, Kyeong-Ho;Heo, Sung-Young;Hong, Seon-Pyo;Lee, Beom-Chan
    • Archives of Pharmacal Research
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    • v.23 no.6
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    • pp.568-573
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    • 2000
  • A reversed-phase high-performance liquid chromatographic method was developed to determine the optical purity of bevantolol enantiomers. (S)-(-)-Menthyl chloroformate((-)-MCF), (S)-(-)-$\alpha$-methylbenzyl isocyanate((-)-MBIC) and 2,3,4,6-tetra-O-acetyl-$\beta$-D-glucopyranosyl isothiocyanate(GITC), which can react with the secondary amine group of bevantolol were investigated as chiral derivatization reagents. Among them indirect chiral HPLC method using CITC gave the best result. The derivatization proceeded quantitatively within 20 min at room temperature. Separation of the enantiomers as diastereomers was achieved by reversed-phase HPLC within 20min using ODS column. Different ratios of (S)-(-)-bevantolol and (R)-(+)-bevantolol were prepared. Enantiomeric separation of these mixtures took place on a chiralcel OD column or, after derivatization with GITC, on a ODS column. No racemization was found during the experiment. This method allowed determination of 0.05% of either enantiomer in the presence of its stereoisomer and method validation showed adequete linearity over the required range.

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Studies on the Separation of Major Bile Acids in Commercial Crude Bile Drugs (시판(市販) 담즙(膽汁) 생약(生藥)중 주요(主要) 담즙산(膽汁酸)의 분리(分離)에 관한 연구(硏究))

  • Park, Jong-Dae;Yoo, Seung-Jo
    • Korean Journal of Pharmacognosy
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    • v.15 no.3
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    • pp.139-146
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    • 1984
  • High performance liquid chromatographic separation is described for the analysis of bile acids after hydrolysis in seven commercial crude bile drugs and ox and pig galls. They are simultaneously separated with HPLC mobile phase of acetonitrile/0.5% ammonium carbonate (pH 6.7) (25.5 : 74.5) at a flow rate $(1.0{\rightarrow}1.5ml/min.)$ and differential refractometer. The linearity of calibration curve and recovery test are good by using the method. The analysis of major bile acids in seven commercial crude bile drugs using the described method is presented. Sample no. 1 of them is similar to separation pattern of ox gall. Sample no. 6 of them is supposed to be genuine bear gall on the basis of identification of ursodeoxycholic acid. Sample $no.\;2{\sim}5$ and 7 of them are supposed to be pig gall on the basis of identification of hyodeoxycholic acid which is a characteristic component of pig gall.

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HPLC SEPARATION AND QUANTITATIVE DETERMINATION OF GINSENOSIDES FROM PANAX GINSENG, PANAX QUINQUEFOLIUM AND FROM GINSENG DRUG PREPARATIONS

  • Soldati F
    • Proceedings of the Ginseng society Conference
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    • 1980.09a
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    • pp.59-69
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    • 1980
  • A new HPLC-method for separation and quantitative determination of ginsenosides in Panax ginseng, Panax quinquefolium and in pharmaceutical drug preparations is elaborated. A reversed-phase-system with ${\mu}Bondapak\;C_{18}$ column (3.9 mm $I.D.{\times}30\;cm$) using acetonitrile-water (30:70) 2 ml/min and acetonitrile-water (18:82) 4 ml/min is suitable for the base-line separation of $Rb_1,\;Rb_2,\;Rc,\;Rd,\;Rf,\;Rg_2,\;respectively\;Re,\;Rg_1$ in 30 minutes. The ginsenosides are directly detected at 203 nm (without derivatization) with the LC-55 or LC-75 spectrophotometer (Perkin-Elmer) at $100\%$ transmission. Detection limit is 300 ng at a signal-to-noise ratio of 10:1. The ginsenosides-peak identification is carried out with HPTLC (high performance thin layer chromatography), with MIR-IR (multiple internal reflection-IR-spectros-copy) and with FD-MS (field desorption mass spectrometry). The calibration curve of each ginsenoside has a correlation coefficient very near to 1. Relative standard deviation for quantitative determinations depends upon the amount of ginsenosides and is approximately 1\%$ for ginsenoside contents of 1\%$. This method is adaptable for routine analysis in quality control laboratories.

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