• Title/Summary/Keyword: HPLC-ELSD detector

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Quantitative Analysis of Platycodin D from Platycodon grandiflorum by HPLC-ELSD (HPLC-ELSD법에 의한 길경의 platycodin D 정량분석)

  • Kim, Geum-Soog;Kim, Hyun-Tae;Seong, Jae-Duck;Park, Hee-Saeng;Kim, Soo-Dong
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.3
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    • pp.200-205
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    • 2002
  • Platycodin D was isolated from n-butanol extract of Platycodi radix(Platycodon grandiflorum and identified by the spectroscopic analysis using $^1H$ and $^{13}C$ NMR techniques. A new method of analysis of platycodin D by high performance liquid chromatography(HPLC) was established using a reversed phase system with YMC-Pack ODS-AM( 250 X 4.6 mm) column and 30% acetonitrile as a mobile phase. Evaporative light scattering detector was used as detector. The kinds of extraction solvents and methods were examined to determine the efficient extraction condition and HPLC analysis was carried out to establish the optimum drying condition for the root of Platycodon grandiflorum. The contents of Platycodin D was highest as 0.083% when platycodon roots were dried at $60^{\circ}C$ using dry oven.

Development and Validation of a Unique HPLC-ELSD Method for Analysis of 1-Deoxynojirimycin Derived from Silkworms (누에에 함유된 1-Deoxynojirimycin의 분석을 위한 HPLC-ELSD 분석법 밸리데이션)

  • Hyejin Cho;Sullim Lee;Myoung-Sook Shin;Joohwan Lee;Sanghyun Lee
    • Korean Journal of Pharmacognosy
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    • v.54 no.1
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    • pp.38-43
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    • 2023
  • A simple and accurate assay was developed for the quantitative analysis of 1-deoxynojirimycin (1-DNJ) derived from the silkworm (Bombyx mori). Normal-phase high-performance liquid chromatography coupled with an evaporative light scattering detector (HPLC-ELSD) and a hydrophilic interaction liquid chromatography column was used. Various parameters were applied to optimize the analysis method. The limits of detection and quantification of 1-DNJ were 2.97 × 10-3 and 9.00 × 10-3 mg/mL, respectively. The calibration curve showed good linearity results. The concentration range and the r2 value were 0.0625-1.0 mg/mL and 0.9997, respectively. The accuracy test demonstrated a significantly high recovery rate (89.95-103.22%). The relative standard deviation was ≤ 1.00%. Thus, a method for the accurate identification and quantitative analysis of 1-DNJ in silkworms was developed. Moreover, in this procedure, the process of derivatization of 1-DNJ, which was required in previous experiments, could be eliminated. This technique may be actively utilized for the development of pharmaceuticals and health functional foods using 1-DNJ.

Determination of Phosphatidylcholine in Korea Functional Foods Containing Lecithins using HPLC with Evaporative Light-Scattering Detector (ELSD) (ELSD를 이용한 레시틴중의 포스파티딜콜린의 분석)

  • Lee Chang-Hee;Bahn Kyeong-Nyeo;Cho Tae-Yong;Lee Ju-Yeon;Lee Young-Ja;Chae Gae Yong
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.267-271
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    • 2005
  • Lecithin is a naturally occurring group of phospholipids found in nearly every living cell and has been widely used as the ingredient of functional foods. Lecithin has high content of phosphatidylcholine(PC), pharmaceutical material which promotes metabolism through the cell membrane. This study was carried out to improve the present inconvenient analytical method of PC in law for health & functional foods. The commodities used in this experiment, were two kinds of egg yolk and eight kinds of soybean lecithin functional foods. PC was separated with isocratic elution with hexane : isopropanol : D.W (30:60:8) through silica column (2.1$\times$150 mm) by HPLC with Evaporative Light-Scattering Detector (ELSD). The flow rate of the eluent was 0.5 ml/mim and infect volume was 10ul. The neubilizer temperature of detector was $60^{\circ}C$, drift tube temperature of that was $75^{\circ}C$ and gas flow was 30 psi. Quantification was carried out by external standardization. Limit of quantification was 0.15ppm. Lecithin contents of egg yolk and soybean Products were > $66\%$ and > $81\%$), respectively. Phosphatidylcholine contents of egg yolk and soybean products were > $74\%$ and > $18\%$, respectively.

A Convenient HPLC/ELSD Method for the Quantitative Analysis of Betaine in Lycium chinense

  • Lee, Sang-Myung;Park, Chae-Kyu;Cho, Byung-Goo;Cho, Kyoung-Shim;Min, Byung-Sun;Bae, Ki-Hwan
    • Natural Product Sciences
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    • v.17 no.2
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    • pp.104-107
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    • 2011
  • In order to facilitate the quality control of betaine from the fruits of Lycium chinense, we have developed a rapid and simple method for quantitative determination. Determination was achieved on a Discovery C18 column with an isocratic solvent system of 0.32% perfluoropentanoic acid aqueous-acetonitrile at a flow-rate of 0.5 mL/min and detected an ELSD. The method was reproducible with intra- and inter-day variations of less than 6% (R.S.D). The recoveries were in the range of 90.01~100.05%. The method turned out to be fast and simple, furthermore, to have a good selectivity and sensitivity for the quantity determination of betaine in the fruits of L.chinense.

Determination and Ethylene Oxide(EO) Separation of Polysorbates by RP-HPLC (역상 HPLC에 의한 polysorbates의 산화에틸렌(EO) 분리 및 정량분석)

  • Lee, Yong-Hwa
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.4
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    • pp.585-593
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    • 2012
  • The EO separation and quantitative determination of polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80 was carried out by reversed phase HPLC. The water/acetonitrile was used for the mobile phase of gradient conditions. An YMC Pack Ph ($250mm{\times}4.6mm$ i.d., $5{\mu}m$) and Phenomenex C4 ($250mm{\times}4.6mm$ i.d., $5{\mu}m$) and the selected ELSD detector was applied. The analysis results of HPLC showed good linearity with correlation coefficient of $r^2$=0.997 in the rage of $180.2{\sim}980.5{\mu}g/mL$ and detection limit.

Determination of Isomaltooligosaccharides in Yoghurts by Using HPLC-ELSD (HPLC-ELSD를 이용한 발효유 제품 중의 Isomaltooligosaccharides 분석법 개발)

  • Ko, Jinhyouk;Lee, Moon-Seok;Kwak, Byung-Man;Ahn, Jang-Hyuk;Park, Jong-Su;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.417-424
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    • 2013
  • A rapid and simple analytical method for the determination of 9 isomaltooligosaccharides (IMO) species in yoghurts was developed using dispersive solid phase extraction (dSPE) clean-up technic and high performance liquid chromatography with evaporative light-scattering detector (HPLC-ELSD). In this study, 9 IMO were extracted from samples simply with chemical reagent using ISO22662 IDF198 method and additional dSPE clean-up. The optimum instrument conditions for the determination were used carbohydrate ES $5{\mu}$ column with gradient elution of water and acetonitrile and ELS detector. The linearity of this method was expressed as the correlation coefficient ($r^2$), the results of IMO 9 species were shown in 0.9999. LOD and LOQ were respectively 7.9-22.1 mg/kg, 25.9-72.8 mg/kg. The accuracy of intra- and inter-day measurements were in the range from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$, and the preceision of the intra- and inter-day measurements were in the range from 0.8 to 7.7%. The recoveries were from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$. The determination results of IMO 9 species for the 9 yoghurts circulated in the market were in the range from $0.317{\pm}0.007$ to $1.624{\pm}0.050$ g/100 g. The newly developed method is appropriate for the determination of IMO in yoghurts, is a rapid and simple method with excellent resolution in compared with previous method.

Sturdy of analysis of Xanthan gum as watersoluble-polymer in cosmetics (화장품 중 수용성 고분자인 Xanthan gum 분석연구)

  • Lee, Yong-Hwa;Yang, Jae-Chan
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.4
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    • pp.664-671
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    • 2013
  • Determination of xanthan gum as watersoluble-polymer in commercial cosmetic samples was carried out by High Perfomance Liquid Chromatography(HPLC). An $C_{18}$ reversed-phase column and the selected ELSD detector was applied. The 25mM ammonium acetate/acetonitrile was used for the mobile phase of gradient conditions. The analysis results of HPLC showed good linearity with correlation coefficient of $r^2=0.9993$ in the rage of $50.3{\sim}604.1{\mu}g/ml$ and detection limit of $12.0{\mu}g/ml$.

Validation on the Analytical Method of Ginsenosides in Red Ginseng

  • Cho B. G.;Nho K. B.;Shon H. J.;Choi K. J.;Lee S. K.;Kim S. C;Ko S. R.;Xie P. S.;Yan Y. Z.;Yang J. W.
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.491-501
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    • 2002
  • A cross-examination between KT&G Central Research Institute and Guangzhou Institute for Drug Control was carried out in order to select optimum conditions for extraction, separation and determination of ginsenosides in red ginseng and to propose a better method for the quantitative analysis of ginsenosides. The optimum extraction conditions of ginsenosides from red ginseng were as follows: the extraction solvent, $70\%$ methanol; the extraction temperature, $100^{\circ}C;$ the extraction time, 1 hour for once; and the repetition of extraction, twice. The optimum separation conditions of ginsenosides on the SepPak $C_{18}$ cartridge were as follows: the loaded amount, 0.4 g of methanol extract; the washing solvents, distilled water of 25 ml at first and then $30\%$ methanol of 25 ml; the elution solvent, $90\%$ methanol of 5 ml. The optimum HPLC conditions for the determination of ginsenosides were as follows: column, Lichrosorb $NH_2(25{\times}0.4cm,$ 5${\mu}m$, Merck Co.); mobile phase, a mixture of acetonitrile/water/isopropanol (80/5/15) and acetonitrile/water/isopropanol (80/20/15) with gradient system; and the detector, ELSD. On the basis of the optimum conditions a method for the quantitative analysis of ginsenosides were proposed and another cross-examination was carried out for the validation of the selected analytical method conditions. The coefficient of variances (CVs) on the contents of ginsenoside-$Rg_{1}$, -Re and $-Rb_1$ were lower than $3\%$ and the recovery rates of ginsenosides were $89.4\~95.7\%,$ which suggests that the above extraction and separation conditions may be reproducible and reasonable. For the selected HPLC/ELSD conditions, the CVs on the detector responses of ginsenoside-Rg, -Re and $-Rb_1$) were also lower than $3\%$, the regression coefficients for the calibration curves of ginsenosides were higher than 0.99 and two adjacent ginsenoside peaks were well separated, which suggests that the above HPLC/ELSD conditions may be good enough for the determination of ginsenosides.

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Mobile Phase Compositions for Ceramide III by Normal Phase High Performance Liquid Chromatography

  • Hong, Seung-Pyo;Lee, Chong-Ho;Kim, Se-Kyung;Yun, Hyun-Shik;Lee, Jung-Heon;Row, Kyung-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.1
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    • pp.47-51
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    • 2004
  • Ceramide III was prepared by the cultivation of Saccharomyces cerevisiae. Ceramide III was partitioned from the cell extracts by solvent extraction and analyzed by Normal Phase High Performance Liquid Chromatography (NP-HPLC) using Evaporative Light Scattering Detector (ELSD). We experimentally determined the mobile phase composition to separate ceramide III with NP-HPLC. Three binary mobile phases of n-hexane/ethanol, n-hexane/lsoprophyl Alcohol(IPA) and n-hexane/n-butanol and one ternary mobile phase of n-hexane/IPA/methanol were demonstrated. For the binary mobile phase of n-hexane/ethanol, the first mobile phase composition, 95/5(v/v), was step-increased to 72/23(v/v) at 3 min. In the binary mobile phase, the retention time of ceramide III was 7.87min, while it was 4.11 min respectively in the ternary system, where the mobile phase composition of n-hexane/IPA/methanol, 85/7/8(v/v/v), was step-increased to 75/10/15(v/v/v) at 3 min. However, in the ternary mobile phase, the more peak area of ceramide III was observed.

Simultaneous Determination of Triterpenoid Saponins from Pulsatilla koreana using High Performance Liquid Chromatography Coupled with a Charged Aerosol Detector (HPLC-CAD)

  • Yeom, Hye-Sun;Suh, Joon-Hyuk;Youm, Jeong-Rok;Han, Sang-Beom
    • Bulletin of the Korean Chemical Society
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    • v.31 no.5
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    • pp.1159-1164
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    • 2010
  • Several triterpenoid saponins from root of Pulsatilla koreana Nakai (Ranunculaceae) were studied and their biological activities were reported. It is difficult to analyze triterpenoid saponins using HPLC-UV due to the lack of chromophores. So, evaporative light scattering detection (ELSD) is used as a valuable alternative to UV detection. More recently, a charged aerosol detection (CAD) has been developed to improve the sensitivity and reproducibility of ELSD. In this study, we developed and validated a novel method of high performance liquid chromatography coupled with a charged aerosol detector for the simultaneous determination of four triterpenoid saponins: pulsatilloside E, pulsatilla saponin H, anemoside B4 and cussosaponin C. Analytes were separated by the Supelco Ascentis$^{(R)}$ Express C18 column (4.6 mm ${\times}$ 150 mm, 2.7 ${\mu}m$) with gradient elution of methanol and water at a flow rate of 0.8 mL/min at $30^{\circ}C$. We examined various factors that could affect the sensitivity of the detectors, including various concentrations of additives, the pH of the mobile phase, and the CAD range. Linear calibration curves were obtained within the concentration ranges of 2 - 200 ${\mu}g$/mL for pulsatilloside E, anemoside $B_4$ and cussosaponin C, and 5 - 500 ${\mu}g$/mL for pulsatilla saponin H with correlation coefficient ($R^2$) greater than 0.995. The limit of detection (LOD) and quantification (LOQ) were 0.04 - 0.2 and 2 - 5 ${\mu}g$/mL, respectively. The validity of the developed HPLC-CAD method was confirmed by satisfactory values of linearity, intra- and inter-day accuracy and precision. This method could be successfully applied to quality evaluation, quality control and monitoring of Pulsatilla koreana.