• Title/Summary/Keyword: HPLC quantification

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Simultaneous Analysis of Cold Medicine Component by High-Performance Liquid Chromatography(HPLC) (고성능 액체크로마토그래피(HPLC)를 이용한 Cold Medicine 성분의 동시 분석)

  • Wonju Lee;Seung-Tae Choi;Keun-Sik Shin;Jin-Young Park;Jae-Ho Sim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.6
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    • pp.867-873
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    • 2023
  • In this study, for the purpose of standardized quality control of a cold medicine, we simultaneous analyzed four main chemical components of a cold medicine: acetaminophen, caffeine, methyl paraben, and propyl paraben. The sample was subjected to quantitative analysis using high performance liquid chromatography (HPLC), after pretreatment of four components. The experiment was carried out by using Isocratic elution at wavelength of 270nm. Acetonitrile and water (H2O) were used as a mobile phase at a flow rate of 1.0mL/min in a commercial C18 reversed-phase column. A volume of 10uL cold medicine were injected into the column with column oven temperature at 35℃. As a result of the experiment, the values of Resolution were 4.983, 1.596, 5.519, and 1.678 respectively-well over Rs >1.5, which indicates that the separation of four components were efficient. In addition, value of symmetry factor of the components was 1.056, 1.069, 1.032, and 1.133 respectively, to show its symmetrical stability. The calibration curve of all four components exhibits good linearity with R2 >0.9995 to 0.9999. Furthermore, the limit of detection(LOD) were between 0.0118 to 1.5973 mg/mL, while the limit of quantification (LOQ) were between 0.0353 to 4.7919 ㎍/mL with the recovery rate of 79.6% ~ 120.5%. The results of this study showed an efficient quality evaluation of a simultaneous analysis method for cold medicine components.

Validation of Precolumn HPLC Oxidation Method for Analysis of Paralytic Shellfish Poison (마비성패류독소 분석을 위한 Precolumn HPLC Oxidation 법의 유효성 검증)

  • Mok, Jong-Soo;Song, Ki-Cheol;Lee, Ka-Jeong;Kim, Ji-Hoe
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.46 no.2
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    • pp.147-153
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    • 2013
  • To prevent paralytic shellfish poisoning (PSP) due to the consumption of shellfish contaminated with PSP toxins, the quantitative analysis of these toxins is very crucial. The AOAC International mouse bioassay (MBA) has been used widely for the routine monitoring of PSP toxins for more than 50 years. However, this method has low sensitivity and high limit of quantification (LOQ) and interferences from other components in the extract, and it cannot determine toxic profiles. Ethical problems also exist with the continued use of this live mouse assay. To establish an alternative method to the MBA used for PSP toxins analysis, we attempted to optimize the analysis conditions of a precolumn high-performance liquid chromatography (HPLC) oxidation method and succeeded in validating its accuracy and precision in quantifying PSP toxins. A clear peak and the isolation of PSP toxins were obtained by injecting the working standards of Certified Reference Materials using HPLC. The LOQ of the precolumn HPLC oxidation method for PSP toxins was about $0.1002{\mu}g/g$, which represented an approximately fourfold improvement in detection capability versus the AOAC MBA. The intra-accuracy and precision for PSP toxins in oysters were 77.0-103.3% and 2.0-5.7%, respectively, while the respective inter-accuracy and precision were 77.3-100.7% and 2.4-6.0%. The mean recoveries of PSP toxins from oysters were 75.2-112.1%. The results of a comparison study showed good correlation between the results of the precolumn HPLC oxidation method and those of MBA, with a correlation factor of 0.9291 for mussels. The precolumn HPLC oxidation method may be used as an alternative to, or supplementary method with, MBA to monitor the occurrence of PSP toxins and to analyze the profiles of these toxins in shellfish.

Simultaneous Quantification of Sulfonamide and Tetracyclines in Fish Muscle Tissue by Matrix Solid Phase Dispersion (MSPD) Extraction and HPLC (MSPD와 HPLC를 이용한 어류의 잔류 설파제와 테트라사이클린계 항생물질의 동시분석)

  • 하대식;김종수;김곤섭
    • Journal of Food Hygiene and Safety
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    • v.12 no.2
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    • pp.117-124
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    • 1997
  • A simple, rapid and simultaneous analytical method is described for the detection of Sulfonamide and Tetracycline residues, i.e., Sulfamerazine (SMR), Sulfamethazine (SMT), Sulfamonomethoxine (SMM), Sulfadimethoxine (SDM), Sulfaquinoxaline (SQN), Oxytetracycline (OXY), Tetracycline (TC), Chlortetracycline (CTC). Blank control and sulfonamide and tetracycline fortified fish muscle samples (0.5 f) were blended with octadecylsilyl (C18, 40 ${\mu}{\textrm}{m}$, 21% load, 60$\AA$) derivatized silica packing material (2 g). Blended fish samples were washed with hexane, then, benzene and dichloromethace were used for the elution of tetracycline and sulfonamide were analyzed by HPLC. Correlation coefficients of standard curves for individual sulfonamide and tetracycline isolated from fortified samples were linear (0.9993$\pm$0.0003~0.9997$\pm$0.0003, 0.9493$\pm$0.078~0.9753$\pm$0.036), respectively. The average percentage recoveries of sulfonamide and tetracycline ranged as 80.86~96.52% to 85.88~92.23%, and 30.01~37.12% to 65.89~73.40%, for the concentration range (0.1~1.0 ppm) examined, respectively. Limit of detection for sulfonamide was 0.0012 ppm for SMR in Paralichthys Olivacleus and 0.0020 ppm for SMR, 0.015 ppm for SMM in Cyprinus Carpio. The applicability of this procedure is demonstrated by separation and detection of incurred tetracycline and sulfonamide residues in fish muscle tissue.

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Simultaneous Determination of Four Bioactive Constituents in Galgeun Tang by HPLC/DAD. (HPLC-DAD를 이용한 갈근탕 중 4종 성분의 동시분석법 확립)

  • Won, Jin-Bae;Jeon, Won-Kyung;Ma, Jin-Yeul;Ma, Choong-Je
    • Korean Journal of Pharmacognosy
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    • v.40 no.3
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    • pp.224-228
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    • 2009
  • For the quality control of traditional herbal medicine, Galgeun tang, simultaneous determination of glycyrrhizin, paeoniflorin, puerarin, 6-gingerol was established by using a high performance liquid chromatographic (HPLC) method with diode array detector. To separate five four constituents, DIONEX $C_{18}$ column ($5{\mu}m$, $120{\AA}$, $4.6\;mm{\times}150\;mm$) was used with gradient elution system of water and methanol. Validation of the chromatography method was evaluated by linearity, recovery, and precision test. Calibration curve of standard components showed excellent linearity ($R^2$>0.9906). Limits of detection (LOD) and limits of quantification (LOQ) varied from 0.15 to $0.52{\mu}g$/ml and 0.27 to $0.80{\mu}mg$/ml, respectively. The relative standard deviations (RSDs) of data of the intra-day and inter-day experiments were less than 2.88% and 1.21%, respectively. The results of recovery test were ranged from 96.71 to 106.29% with RSD values 0.01-0.80%.

Lignan contents in Acanthopanax senticosus by HPLC (고속액체크로마토그래피를 이용한 가시오갈피의 리그난 함량)

  • Kim, Hye-Min;Cho, Seon-Haeng;Kook, Soon-Ja;Lee, Sang-Hyun
    • Korean Journal of Agricultural Science
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    • v.38 no.3
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    • pp.479-484
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    • 2011
  • A reverse-phase system of HPLC using a linear gradient of acetonitrile and deionized water was developed for the quantification lignans, eleutherosides B and E, in Acanthopanax senticosus. The HPLC system consisted of linear gradient of acetonitrile and deionized water, and UV/VIS detection was set at 210 nm. Both eleutherosides B and E contents in different parts of A. senticosus were determined. As a result, the contents of eleutherosides B and E were measured in the leaves (trace amounts and 0.029 mg/g, respectively), stems (0.107 and 1.015 mg/g, respectively), roots (0.026 and 0.390 mg/g, respectively), and fruits (0.022 and 0.043 mg/g, respectively). Moreover, eleutherosides B and E in the water extract were found 0.011 and 0.171 mg/g, respectively.

The Use of the Online Two-dimensional Liquid Chromatography Coupled with a Universal Detector for the Screening of Non-volatile Potential Migrants in Food Packaging Materials (식품포장재내 비휘발성 잠재 이행물질들의 스크리닝을 위한 이차원크로마토 그래피와 범용검출기의 이용)

  • Yoon, Chan-Suk;Lee, Keun-Taik
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.16 no.1
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    • pp.9-18
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    • 2010
  • For screening test of the non-volatile compounds which migrate from food packaging materials into foodstuffs, the traditional high performance liquid chromatography (HPLC) systems suffer from the lack of universal detector with high sensitivity and universality and high efficiency HPLC separation column which provides complete separation of complex mixtures into all individual substances. In this work, the use possibility of online two-dimensional liquid chromatography (2D-LC) system coupled with a charged aerosol detector (CAD), a universal detector, was reviewed. 2D-LC system permits to improve peak capacity and resolving power for complex mixtures. Charged aerosol detector (CAD) offers a new feasibility for detection of any non-volatile compounds with high sensitivity and constant response factor in a calibration range. The combination of size exclusion chromatography (SEC) and normal phase HPLC (NP-HPLC) is most frequently used for the separation of the natural and synthetic polymers which are mainly used as raw materials for the manufacture of food packaging materials. However, there is no commercial software available for data acquisition and handling and therefore the quantification in 2D-LC analysis is still rare.

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The Effective Preparation of Flavonoids from Scutellaria baicalensis GEORGI by Diaion HP-20 Resin

  • Yu, Young-Beob
    • Korean Journal of Plant Resources
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    • v.27 no.6
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    • pp.635-641
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    • 2014
  • Scutellaria baicalensis $G_{EORGI}$ (Scutellariae Radix) has been used to clear heat and to dry dampness in the stomach or intestines, which manifests as diarrhea or dysenteric disorder. In this study, we investigated the effective preparation of active components in Scutellariae Radix using the methods of solvent extraction and absorption fractionation for the development of new functional food or pharmaceuticals. The marker substances, baicalin, baicalein, wogonoside, and wogonin were directly isolated from the Scutellariae Radix. There chemical structures were elucidated by spectroscopic analysis. The Scutellariae Radix was extracted with hot water. To enhance yield of flavonoids in Scutellariae Radix, the hot water extract was dissolved in ethanol with concentration dependent manner. The precipitates were separated using centrifugal techniques at 10,000 rpm. Supernatant liquid was applied to the HPLC for quantification of major compounds. Separately, the hot water extract was absorbed on Diaion HP-20 resin. And then, the absorbed fraction was eluted with methanol for HPLC. The contents of baicalin, baicalein, wogonoside and wogonin in different treatment methods were analyzed by HPLC. Total amount of four major components were 16.9% in 50% ethanol extract, 21.7% in 70% ethanol extract, 20.5% in 90% ethanol extract, and 39.3% in absorbed fraction of Diaion HP-20 resin. In these results, we found that resin absorption method is suitable for the extraction of enriched flavonoids from Scutellariae Radix.

Quantitation of Mevinolinic Acid in Human Plasma by HPLC (고속액체크로마토그래피를 이용한 사람 혈장중 메비놀린 산의 정량)

  • Oh, Han-Suk;Park, Dong-Young;Seo, Sung-Hoon;Kim, Young-Gwan;Hong, Seon-Pyo;Choi, Young-Wook;Lee, Kyung-Tae
    • Journal of Pharmaceutical Investigation
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    • v.30 no.4
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    • pp.279-282
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    • 2000
  • Simple and precise high-performance liquid chromatographic (HPLC) assay was developed and validated for the determination of a HMG-CoA reductase inhibitor, $lovastatin^{TM}$ and its active metabolite (mevinolinic acid) in human plasma. The method involved solid phase extraction of mevinolinic acid and internal standard using Sep-Pak Cartridge. Samples were analyzed by reversed-phase HPLC using $Capcell-Pak\;C_{18}$ column with ultraviolet detection at 238 nm. The quantitation limit of mevinolinic acid was 2 ng/ml and the calibration curve was linear over the range of 2-50 ng/ml $(r^2>0.999)$ with human plasma. The analyses of quality control samples indicated that the normal values could be predicted with an accuracy >97%. The intra- and inter-day coefficients of variation for the analyses were <10%. The average recoveries were similar (79%) for mevinolinic acid and methylmevinolinic acid. The method described has been successfully applied to the quantification of mevinolinic acid in about 1,000 human plasma samples over six-month period.

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Validation of a Simple HPLC Method for Determination of Ciprofloxacin Hydrochloride in Human Plasma (인체 혈장에서 염산시프로플록사신(시프로플록사신으로서 250 mg) 정량을 위한 HPLC 분석법의 유효성검토)

  • Ha, Yong-Hwa;Cho, Sung-Hee;Chun, Sung-Kuk;Seo, Seong-Hoon;Rew, Jae-Hwan;Choi, Young-Wook;Lee, Kyung-Tae
    • Journal of Pharmaceutical Investigation
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    • v.34 no.4
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    • pp.327-331
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    • 2004
  • A simple HPLC method with ultraviolet detection of ciprofloxacin in human plasma was developed and validated. After protein precipitation with trichloroacetic acid, chromatographic separation of ciprofloxacin in plasma was achieved at $50^{\circ}C$ with a $C_{18}$ column and methanol-phosphate mixture (pH 2.5), as mobile phase. Quantitative determination was performed by ultraviolet detection at 278 nm. The method was specific and validated with a limit of quantification of 100 ng/ml. The intra- and inter-day coefficients of variation were between 1.67% and 10.55% and accuracy between 92.01 % and 106.09%. The method has been successfully applied in a bioavailability study of 250 mg ciprofloxacin hydrochloride tablet.

Antioxidant Activity of Vitex rotundifolia Seeds and Phytochemical Analysis Using HPLC-PDA

  • Hyejin Cho;Hak-Dong Lee;Jae Min Chung;Sanghyun Lee
    • Natural Product Sciences
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    • v.29 no.1
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    • pp.1-9
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    • 2023
  • This study assessed in vitro antioxidant activity (ABTS+ and DPPH) of Vitex rotundifolia seeds collected from two different regions in Korea (Jungjang City and Sindu City). Three extraction methods using ethanol, methanol, and water were prepared separately and subjected to quantification by reverse-phase high-performance liquid chromatography-photodiode array (HPLC-PDA) analysis as well as antioxidant testing. Among them, the water-based extract exhibited superior activity in the ABTS+ compared with the ethanol- and methanol-based extracts, while the DPPH assay analysis, revealed that the methanol-based extract had very low antioxidant activity. The concentrations of vanillic acid (1), luteolin (2), vitexicarpin (3), and artemetin (4) were quantified using HPLC-PDA analysis. Vanillic acid (1) was identified as the main antioxidant in V. rotundifolia seeds. Combining the antioxidant activity and quantitative analysis results, the water-based extract was considered to have the highest antioxidant activity. Furthermore, vanillic acid (1) was detected in the leaves and stems of V. rotundifolia plants from different regions, indicating that this species has the potential for use in future antioxidant-applications.