• Title/Summary/Keyword: high performance liquid chromatography

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Mutation Analysis in β2-Adrenergic Receptor Gene by Single Strand Conformation Polymorphism (SSCP) and Denaturing High Performance Liquid Chromatography (DHPLC) (SSCP와 DHPLC에 의한 β2-교감신경수용체 유전자의 돌연변이 분석)

  • Park, Sang-Bum;Han, Sang-Man;Nam, Youn-Hyoung;Jang, Won-Cheoul
    • Analytical Science and Technology
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    • v.17 no.1
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    • pp.53-59
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    • 2004
  • Up to now, methods for the detection of genetic alterations as single strand conformation polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE) have been used. It is too labor-intensive and expensive to serve for routine analysis. Moreover, lower in its sensitivity and specificity being also strongly dependent on the experience of the investigater. To improve these problems, we analysed mutation of ${\beta}_2$-adrenergic receptor gene that controls bronchial asthma by denaturing high performance liquid chromatography (DHPLC) according to ion-pair reversed phase chromatography (IP-RPC). We extracted genomic DNA from 80 asthma patients and then amplified DNA using PCR and analysed PCR product by SSCP and DHPLC. As a result, we analysed mutation frequency is 19 (23.75%) on SSCP and 25 (31.25%) on DHPLC in ${\beta}_2$-adrenergic receptor gene. We conclude that DHPLC is a fast and simple screening method rather than SSCP analysis.

A Study on the Determination of Caffeine in Coffee, Black tea and Green Tea by high performance Liquid Chromatography (고속액체 크로마토크래피에 의한 커피, 홍차, 녹차중의 카페인 정량에 관한 연구)

  • 권익부;이윤수;우상규;이충영;서준걸
    • Journal of Food Hygiene and Safety
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    • v.5 no.4
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    • pp.213-217
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    • 1990
  • A simple and practical method for the determination of caffeine in coffee, black tea and green tea was studied. The analysis of caffeine was performed by reverse phase high perfomance liquid chromatography using a ${\mu}-Bondapak$ C18 column at isocratic condition with methanol-acetic acid-water (20: 1: 79) on UV detector at 280 nm. The extraction and clean-up of caffeine in sample is based on combing a simple pretreatment with the use of a Sep-Pak Alumina A cartridge. The average recoveries of caffeine from several samples were 95.2 -101.3%, the relative standard deviation for the whole procedure was 0.10 ~ 0.62%, and the detection limit of caffeine in sample solution was about $0.1\;\mu\textrm{g}\;per\;ml$.

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Simultaneous Determination of Creatine, Dicyandiamide and Dihydrotriazine in Dietary Supplements by High Performance Liquid Chromatography (고성능 액체크로마토그래피를 이용한 식이보충제에서 크레아틴, 디시안디아마이드, 디하이드로트리아진의 동시분석)

  • Park, Sang-Wook;Yoo, Myung-Sang;Lee, Wonjae
    • KSBB Journal
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    • v.29 no.4
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    • pp.232-238
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    • 2014
  • The simultaneous determination of creatine monohydrate (CrM), dicyandiamide and dihydrotriazine in dietary supplements using reversed-phase high performance liquid chromatography (HPLC) was developed. Chromatography was performed on a Nuclosil 100-5 SA ($4.6{\times}250mm$) column with a mobile phase of 2.3% ammonium phosphate (pH 5.5), and UV detection at 224 nm, 212 nm, and 237 nm, respectively. The performance characteristics of HPLC were determined in terms of selectivity, linearity, precision, recovery, limit of detection (LOD), and limit of quantification (LOQ). The calibration curves were linear within the concentration range of $40.0{\sim}500.0{\mu}g/mL$ for creatine, $0.1{\sim}12.8{\mu}g/mL$ for dicyandiamide, and $0.05{\sim}6.4{\mu}g/mL$ for dihydrotriazine. The detection limits of the method were 1.09, 0.01, and $0.08{\mu}g/mL$ for creatine, dicyandiamide, and dihydrotriazine, respectively. The recoveries of creatine, dicyandiamide, and dihydrotriazine were 97.2~100.9, 92.3~106.5, and 97.2~105.5%, respectively. It is expected that the chromatographic analytical method developed in this study will be usefully applicable to simultaneous determination of creatine, dicyandiamide, and dihydrotriazine contained in dietary supplements.

Simultaneous Determination of Sulfonamides in Porcine and Chicken Muscle Using High Performance Liquid Chromatography with Ultraviolet Detector

  • Shim, You-Sin;Shin, Dong-Bin;Cho, Yong-Sun;Choi, Yun-Hee;Lee, Sang-Hee
    • Food Science and Biotechnology
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    • v.18 no.6
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    • pp.1430-1434
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    • 2009
  • The present study used the liquid extraction pretreatment method and developed a liquid chromatographyultraviolet detector (LC-UV) for the simultaneous determination of 14 sulfonamides (SAs) residues in porcine and chicken muscle. Linearity within a range of $50-150\;{\mu}g/kg$ was obtained with the correlation coefficient ($r^2$) of 0.9673-0.9997. The mean recovery of SAs was 55.9-109.7% (relative standard deviations; RSDs 1.7-17.3%) in porcine muscle and 52.8-112.4% (RSDs 2.3-16.9%) in chicken muscle. The limits of detection (LODs) and limits of quantification (LOQs) were 2-32 and $7-96\;{\mu}g/kg$ in porcine muscle, and 4-32 and $13-97\;{\mu}g/kg$ in chicken muscle, respectively. These values were lower than the maximum residue limits (MRLs) established by the European Union. The sum of all SAs residues present should be less than $100\;{\mu}g/kg$.

Rapid Isolation of Cyanidin 3-Glucoside and Peonidin 3-Glucoside from Black Rice (Oryza sativa) Using High-Performance Countercurrent Chromatography and Reversed-Phase Column Chromatography

  • Jeon, Heejin;Choi, Janggyoo;Choi, Soo-Jung;Lee, Chang Uk;Yoon, Shin Hee;Kim, Jinwoong;Yoon, Kee Dong
    • Natural Product Sciences
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    • v.21 no.1
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    • pp.30-33
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    • 2015
  • Anthocyanins are water soluble plant pigments which are responsible for the blue, red, pink, violet colors in several plant organs such as flowers, fruits, leaves and roots. In recent years, anthocyanin-rich foods have been favored as dietary supplements and health care products due to diverse biological activities of anthocyanins including antioxidant, anti-allergic, anti-diabetic, anti-microbial, anti-cancer and preventing cardiovascular disease. High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase medium pressure liquid chromatography (RP MPLC) method was applied for the rapid and efficient isolation of cyanidin 3-glucoside (C3G) and peonidin 3-glucoside (P3G) from black rice (Oryza sativa L., Poaceae). The crude black rice extract (500 mg) was subjected to HPCCC using two-phase solvent system composed of tert-butyl methyl ether/n-butanol/ acetonitrile/0.01% trifluoroacetic acid (TBME/B/A/0.01% TFA, 1 : 3 : 1 : 5, v/v, flow rate - 4.5 mL/min, reversed phase mode) to give enriched anthocyanin extract (37.4 mg), and enriched anthocyanin extract was sequentially chromatographed on RP-MPLC to yield C3G (16.5 mg) and P3G (8.7 mg). The recovery rate and purity of isolated C3G were 76.0% and 98.2%, respectively, and those of P3G were 58.3% and 96.3%, respectively. The present study indicates that HPCCC coupled with RP-MPLC method is more rapid and efficient than multi-step conventional column chromatography for the separation of anthocyanins.

Analysis of Vicamine Using High Performance Liquid Chromatography and Antioxidant Activity of Vincaminor Extract (High performance liquid chromatography를 이용한 빈카민 분석 및 빈카마이너의 항산화능 측정)

  • Jung, Jong-Hee;Back, Yu-Mi;Lee, Kwang-Geun
    • Korean Journal of Food Science and Technology
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    • v.40 no.5
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    • pp.599-602
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    • 2008
  • Vincamine, one of the major indole alkaloids in vincaminor (Vinca minor L.) is commonly used for treating cerebrovascular diseases. The antioxidant activity of vincaminor extracts and vincamine were measured by 1.1-diphenyl-2-picrylhydrazyl (DPPH) and lipid malonaldehyde (MA) assay. Vincaminor leaves were pulverized and extracted with various solvents such as water, methanol, and ethanol. The antioxidant activities of the extracts varied in accordance with solvents and assays. In DPPH assay, the water extract showed the highest antioxidant activity. In lipid MA assay, However, the ethanol extract inhibited MA formation from cod liver oil by 82% at the level of 5,000 ${\mu}g/mL$. Vincamine in the extract was analyzed by high-performance liquid chromatogram and the concentration of vincamine was 0.419$\pm$0.005 ${\mu}g/mL$.

Marker compounds contents of Salvia miltiorrhiza Radix depending on the cultivation regions

  • Seong, Gi-Un;Kim, Mi-Yeon;Chung, Shin-Kyo
    • Journal of Applied Biological Chemistry
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    • v.62 no.2
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    • pp.129-135
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    • 2019
  • Salvia miltiorrhiza Radix is cultivated in Korea and China and is traditionally used to treat cardiovascular diseases. In this study, we developed and validated a quantitative analysis method for S. miltiorrhiza Radix using high-performance liquid chromatography (HPLC). Identification was performed using ultra performance liquid chromatography-tandem mass spectrometry. For quantitative analysis, we used seven marker compounds. Separation conditions for HPLC were optimized using an ODS column with gradient conditions of 1% formic acid in distilled water and 1% formic acid in acetonitrile, with a flow rate of 0.8 mL/min and a detection wavelength of 280 nm. This method showed good linearity ($R^2=0.9998$), precision (relative standard deviation ${\leq}3.3%$), accuracy (recovery of 94.16-102.89%), limit of detection ($7.53{\mu}g/mL$), and limit of quantification ($23.71{\mu}g/mL$). This approach successfully quantified marker compounds in S. miltiorrhiza Radix. The individual marker compounds were identified by comparing the molecular masses and retention times with does standard compounds. Marker compound contents of S. miltiorrhiza Radix were investigated with different cultivation regions. Seven marker compounds were detected and quantified in all samples. Among them, salvianolic acid B showed the highest contents and it ranged from 4.13 to 7.15%. The salvianolic acid B content (7.15%) of marker compound was the highest in Bonghwa, and the tanshinone IIA content (1.90%) was the highest in Pohang. The results of marker compounds and developed method were intended to provide a favorable reference for the study of S. miltiorrhiza Radix from different regions of Korea.

The Application of High Performance Thin Layer Chromatography for Herbal Formula Standardization (한약처방의 표준화를 위한 HPTLC의 활용)

  • Choi, Min-Kyung;Kim, Hyeong-Geug;Wang, Jing-Hua;Son, Chang-Gue
    • The Journal of Korean Medicine
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    • v.32 no.4
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    • pp.68-74
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    • 2011
  • Objective: The purpose of this study is to expatiate on high performance thin layer chromatography (HPTLC) as a simple, easy and scientific method for evaluation and standardization of herbal formulae. Methods: Through retrieving HPTLC application for herbal formulae in the literatures, the current situation of HPTLC application, and potential as well as limitation of HPTLC as a standardization method for multi-herbal drug was studied. Results: HPTLC is a speedy, inexpensive and well-operable tool for possessing multi-capability on component identification, separation, quantification and purification compared to other methods, such as high performance liquid chromatography (HPLC), gas chromatography (GC). Conclusions: HPTLC is considered as an available and convenient method for quality control and standardization of multi-herbal drugs. Thereby, this method could be recommended to widely applicate in the traditional Korean medicine.

Simultaneous detection for synthetic antimicrobials in muscle by high performance liquid chromatography-mass selective detector (HPLC-MSD) (HPLC-MSD 를 이용한 식육 중 합성항균제의 동시분석)

  • Hong In-Suk;Choi Yoon-Hwa;Kwon Taek-Boo;Lee Jung-Hark
    • Korean Journal of Veterinary Service
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    • v.29 no.3
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    • pp.317-330
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    • 2006
  • This study was conducted to develop the analytical method about simultaneous determination for synthetic antimicrobials in muscle by high performance liquid chromatography - mass selective detector (HPLC- MSD). Solid phase extraction (SPE), matrix solid phase dispersion (MSPD) and liquid-liquid extraction (LLE) have been adapted as pretreatment procedures for HPLC- MSD. Among various solvent tested, methanol was chosen for extraction of synthetic antimicrobials in muscles. For the optimized response, the values of various MS parameters including fragment voltage, drying gas flow, nebulizer pressure, drying gas temperature were verified. The average recovery rates using MSPD and SPE for muscles of bovine and pork were 78.9-127.1% and 78.3-121.7%, respectively. This method was verified the satisfactory performance for fourteen synthetic antimicrobials excepting carbadox in muscle of pork as detection limit of $0.05{\mu}g/g$ on API/ES SIM mode.