• 제목/요약/키워드: high-performance liquid chromatography (HPLC)

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HPLC-RTS (high performance liquid chromatography-retention time shift)를 이용한 암모늄 이온의 안정동위원소 측정방법의 개선 (Stable Isotope Measurement of Ammonium Using HPLC-RTS (high performance liquid chromatography-retention time shift))

  • 안순모;이지영
    • 한국해양학회지:바다
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    • 제18권1호
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    • pp.47-52
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    • 2013
  • 생태계 질소 순환 연구에서 동위원소 추적자를 이용한 실험은 그 유용성에도 불구하고, 무기 질소의 안정동위원소 측정이 어려워 자료의 생산이 활발하지 못한 형편이다. Gardner et al. (1996)은 HPLC (high performance liquid chromatography)를 이용하여 비교적 간편하고 정밀하게 암모늄의 동위원소 비를 측정하는 방법을 개발해 질소순환 연구에 활발하게 활용하였다. 그러나 일반적이지 않은 syringe 펌프가 필요해 다른 실험실에 이 시스템이 도입되지는 못하였다. 본 연구에서는 최신 HPLC 펌프 기술을 활용하여 Dr. Gardner 그룹 이후 최초로 암모늄 동위원소비 측정 시스템을 구축하였다. 표준시료 측정 결과 RTS (retention time shift)는 안정동위원소 비와 좋은 상관관계를 보였다. 이 시스템은 측정 자료를 디지털 형태로 변환하는 장치까지 갖추어 컴퓨터 프로그램을 이용한 다양한 자료처리가 가능하다. 특히 Matlab$^{(R)}$ 이용하여 retention time 계산에 peak 면적을 이용함으로써 측정의 정확성을 높이고, 직선성을 향상할 수 있었다. 영산강 하구 해수를 가지고 실시한 $^{15}N$-암모늄 첨가 실험에서 $^{15}N/^{14}N$ 비율 변화를 HPLC-RTS법을 통해 측정하였으며, 식물성 플랑크톤에 의한 암모늄 섭취와 재광물화 과정을 각각 독립적으로 측정하는 것이 가능했다.

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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    • 제9권1호
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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 Glutamate, Glycine, and Alanine in Human Plasma Using Precolumn Derivatization with 6-Aminoquinolyl-N-hydroxysuccinimidyl Carbamate and High- Performance Liquid Chromatography

  • Li, Qing Zhong;Huang, Qing Xian;Li, Shu Cui;Yang, Mei Zi;Rao, Bin
    • The Korean Journal of Physiology and Pharmacology
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    • 제16권5호
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    • pp.355-360
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    • 2012
  • A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method has been validated for determining concentrations of glutamate, glycine, and alanine in human plasma. Proteins in plasma were precipitated with perchloric acid, followed by derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Simultaneous analysis of glutamate, glycine, and alanine is achieved using reversed-phase HPLC conditions and ultraviolet detection. Excellent linearity was observed for these three amino acids over their concentration ranges with correlation coefficients (r)>0.999. The intra- and inter-day precision were below 10%. This method utilizes quality control samples and demonstrates excellent plasma recovery and accuracy. The developed method has been successfully applied to measure plasma glutamate, glycine, and alanine in twenty volunteers.

Quantitative and Qualitative Analysis of Alkaloids in Coptis chinensis (Coptidis Rhizoma) by LC-DAD and LC-ESI/MS

  • Yu, Young-Beob;Bae, Chang-Hyu
    • 한국자원식물학회지
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    • 제30권6호
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    • pp.693-698
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    • 2017
  • The quality control of natural products is principal key to guarantee the Good Manufacturing Practices (GMP) and Good Clinical Practices (GCP) for the functional food, pharmaceuticals and cosmeceuticals in the industry. In this study, we examined the quantitative analysis of berberine as marker substance of Coptidis Rhizoma by high performance liquid chromatography-photodiode array detector (HPLC-DAD). The HPLC method was validated and met all the requirements for the quality control analysis recommended by FDA and ICH. The berberine was separated on a Xterra $C_{18}$ column ($5{\mu}m$, $4.6{\times}250mm$) using mobile phase consisting of distilled water and acetonitrile with $KH_2PO_4$ (3.4 g) and $Na_2SO_4$ (1.7 g). Calibration curve of berberine has been estimated (y = 42293.47x-41589 with the correlation coefficient 0.9999). The amount of berberine was calculated as 4.25%. And berberastine, palmatine, columbamine, jatrorrhizine, epiberberine, berberine and coptisine in the Coptidis Rhizoma were identified by high performance liquid chromatography - electrospray ionization-mass spectrometer (HPLC-ESI-MS) method.

LC/MS에 의한 원료생강 및 생강 페이스트 중의 Gingerol 화합물 분석 (Analysis of Gingerol Compounds of Raw Ginger (Zingiber officinale Roscoe) and Its Paste by High Performance Liquid Chromatograph-Mass Spectrometry (LC/MS))

  • 조길석
    • 한국식품영양과학회지
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    • 제29권5호
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    • pp.747-751
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    • 2000
  • 본 연구에서는 생각의 매운맛을 주요성분인 gingerol 화합물의 분석방법을 개선할 목적으로, 원료 생각을 아세톤으로 추출한 후 그 추출물을 TLC 및 LiChrosorb RP-18 column으로 장착된 HPLC를 사용하여 3가지 gingerol 동족체 화합물을 분리하였고, LC/MS 및 NMR로 확인한 결과 생강의 동족체 화합물은 [6]-, [8]-, [10]-gingerol로 확인되었다. 본 방법으로 추출한 gingerol 표준품을 사용하여 시판 원료생강 및 생각 페이스트 중의 gingerol 함량을 측정한 결과, 전자의 경우 [6]-, [8]-, [10]-gingerol 함량은 635.5 mg%, 206.6 mg%, 145.7 mg%이었고, 후자의 경우는 418.2 mg%, 142.6 mg%, 103.3 mg%로 각각 구성되어 있었다.

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Simple and Sensitive Determination of Baclofen in Human Plasma by Column-Switching and Semi-Micro High-Performance Liquid Chromatography

  • Ban, Eun-Mi;Ko, Hye-Ran;Kim, Chong-Kook
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.284.2-284.2
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    • 2003
  • Using a column-switching technique. highly sensitive and selective semi-micro high-performance liquid chromatographic (HPLC) method has been developed for the determination of baclofen in human plasma. Following precipitation of plasma sample containing baclofen with zinc sulfate-acetonitrile, samples were directly injected on to the system. (omitted)

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소변 중 뮤콘산의 친환경 HPLC 분석방법을 이용한 벤젠 노출 생물학적 노출평가 (Environmentally friendly determination of urinary trans, trans-muconic acid for biological monitoring of benzene exposure by green high-performance liquid chromatography)

  • 양동혁;이미영;원용림
    • 분석과학
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    • 제25권6호
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    • pp.460-466
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    • 2012
  • 벤젠 노출의 생물학적 노출인 인체 소변 중 뮤콘산(trans, trans-muconic acid) 분석을 위한 녹색 HPLC (Highr performance liquid chromatography) 분석방법을 개발하고 검증하였다. 이온교환수지 고상추출 카트리지를 사용하여 소변 중 뮤콘산을 추출하고, MF-Ph1 SG80 ($150mm{\times}2.0mm$ I.D., 5 ${\mu}m$) 칼럼과 이동상(10 mM $KH_2PO_4$+0.1% $H_3PO_4$)을 뮤콘산의 HPLC 분석에 사용하였으며 검출은 자외가시부검출기 259 nm에서 실시하였다. 0.1-5.0 mg/L 범위에서 상관계수 ($r^2$) 0.9992 이상의 양호한 직선성을 나타내는 검량선을 얻었다. 일내 및 일간 정밀성(RSD)은 각각 0.9-8.5% 및 3.1-4.5%였으며, 평균 회수율은 97.5-101.7% 범위였다. 본 연구에서는 유기 용매를 전혀 사용하지 않지만 기존 방법과 동일한 분석 성능을 가지고 있는 친환경 분석법을 개발하였다. 확립된 분석법은 벤젠 노출 평가를 위한 근로자의 생물학적 모니터링 방법으로 활용될 수 있을 것이다.

Phytochemical constituents of Lactuca serriola leaves and their content analysis by HPLC-UV

  • Kim, Juree;Lee, Hak-Dong;Choi, Jungwon;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • 제65권3호
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    • pp.153-158
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    • 2022
  • This study aimed to identify the phytochemical constituents of Lactuca serriola leaves and perform quantitative analysis of the methanol (MeOH) extract of L. serriola, L. indica, L. raddeana, L. sativa, and L. triangulata. Six compounds were isolated from the MeOH extracts of L. serriola using open column chromatography and identified as protocatechuic acid (1), caffeic acid (2), cynaroside (3), apigenin 7-glucuronide (4), luteolin (5), and apigenin (6) using 1H-, 13C-nuclear magnetic resonance, and mass spectrometry. Quantitative analysis of the six compounds was performed on the MeOH extract of Lactuca species using high-performance liquid chromatography (HPLC) and an ultraviolet (UV). A reverse-phased column was used, and the UV absorbance was set to 280 nm. The contents of compounds 2 and 3 were the highest (1.58 and 2.64 mg/g ext., respectively) in L. serriola extracts. However, compounds 4 and 6 were higher (1.46 and 0.40 mg/g ext., respectively) in L. triangulata. These results provide quantitative data for the application of Lactuca species in the pharmaceutical and functional food industries.

Simultaneous determination of bisphenol A, chlorophenols and alkylphenols by solid-phase extraction and HPLC

  • Lee, Taejoon;Park, Keun-Young;Pyo, Dongjin
    • 분석과학
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    • 제30권1호
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    • pp.20-25
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    • 2017
  • An analytical method for determining potential endocrine disruptors (bisphenol A, 2-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, pentachlorophenol, p-t-butylphenol, p-pentylphenol, p-hexylphenol, p-t-octylphenol, p-heptylphenol, nonylphenol) by solid-phase extraction (SPE) and High Perfomance Liquid Chromatography(HPLC) equipped with fluorescence and variable wavelength detector has been developed. The SPE process for sample concentration was performed on a commercially available Oasis HLB cartridge packed with polymeric sorbents. The effect of elution solvent and elution volume on the recoveries of the analytes were investigated with HPLC. Average recovery of >85% was achieved with 60mg sorbents using 5mL of methanol as elution solvent. Phenolic compounds in canned drinks, beverages and water samples were surveyed by this proposed method.

고속액체크로마토그라피에 의한 은행잎중 Flavonoid Glycoside의 확인 및 정량 (Identification and Quantitative Analysis of Flavonol Glycosides from Ginkgo biloba Leaves by High Performance Liquid Chromatography)

  • 강삼식;김주선;곽의종;김기협
    • 생약학회지
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    • 제21권2호
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    • pp.148-152
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    • 1990
  • Seven flavonol glycosides from the EtOAc fraction of Ginkgo biloba leaves were identified by high performance liquid chromatography. Separation by reversed phase chromatography on $Lichrosorb^{\circledR}$ RP-18 column was achieved by isocratic elution. The content of the major acylated flavonol glycoside, kaempferol 3-0-[$6'-O-{p}-coumaroyl-{\beta}-_D-glucosyl(1{\rightarrow}2)-{\alpha}-_L-rhamnoside$] was about 8.0% and 0.55% for EtOAc fraction and MeOH extract, respectively.

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