• Title/Summary/Keyword: HPLC detector

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Determination of Catecholamines and Their Metabolites in Rat Brain by High Performance Liquid Chromatography with Electrochemical Detector (HPLC-ECD에 의한 흰쥐 뇌 부위별 Catecholamine 및 대사산물의 신속정량법)

  • Ro, Ihl-Hyeob
    • YAKHAK HOEJI
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    • v.32 no.1
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    • pp.50-54
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    • 1988
  • A simple and sensitive method was studied for the simultaneous determination of catecholamine, indoleamine and their related metabolites by high performance liquid chromatography with electrochemical detector. Norepinephrine, dopamine, serotonin and their metabolites of 3,4-dihydroxyphenylacetic acid, homovanillic acid, 5-indoleacetic acid were resolved from rat brain tissue homogenates by separation on reversed phase $C_{18}$ column with mobile phase consisting of monochloroacetate buffer (pH2.47), 1.42mM sodium octyl sulfonate and 7% acetonitrile. Both catechols and indoles can be eluted in 15min. The sensitivities of this method are sufficient for determination of at least 100 pg of neurochemical amines in brain samples, for example, frontal cortex, olfactory bulb, striatum, septum, hippocampus, thalamus, hypothalamus, medulla & pons and cerebellum. The highest level of dopamine was observed in striatum whereas norepinephrine and serotonin were in hypothalamus.

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Discrimination between steam processed and unprocessed Tubers of Gastrodia elata Blume by HPLC

  • Zhao, Bing Tian;Song, Si Whan;Le, Duc Dat;Ma, Eun Sook;Son, Jong Keun;Woo, Mi Hee
    • Analytical Science and Technology
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    • v.32 no.6
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    • pp.217-224
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    • 2019
  • In this study, to evaluate the effectiveness and safety of oral therapy using Gastrodiae Rhizoma, a new HPLC-PDA analysis method was developed for the simultaneous quantitation of the three major components: (1) gastrodin, (2) gastrodigenin, and (3) p-hydroxybenzaldehyde, in steam processed and unprocessed tubers of Gastrodia elata Blume. The clear separation of the three components was achieved on a C18 column (250 × 4.6 mm, 5 ㎛) by gradient elution using water (including 0.1 % formic acid) and acetonitrile as the mobile phase. The flow rate was 1.0 mL/min, and the UV detector wavelength was set at 270 nm. The results demonstrate satisfactory linearity, recovery, precision, accuracy, stability, and robustness. The established HPLC-PDA method was applied to quantify three major compounds in 59 samples of G. elata Blume tubers. Finally, the steam processed and unprocessed tubers of G. elata Blume were successfully distinguished by pattern recognition analysis.

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.

Determination of eugenol in Eugenia caryophyllata by high-performance Liquid chromatography with photodiode array detection and method validation (HPLC-PDA를 이용한 정향(Eugenia caryophyllata) 중의 eugenol 분석법 확립 및 검증)

  • Yun, Hyeong-Jun;Yun, So-Mi;Lee, Myoung-Heon;Son, Seong-Wan
    • Korean Journal of Veterinary Research
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    • v.48 no.1
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    • pp.9-16
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    • 2008
  • A method for the quantification of eugenol in the medicinal herb Clove was developed and validated. For preparation of sample solutions clove was dried at $60^{\circ}C$ for 2h and ground by mixer and extracted with 95% ethanol for shaking extraction. The elutes were analyzed by HPLC system included a reversed phase column, a isocratic mobile phase of 60% methanol and PDA detector set at 280 nm. Calibration graphs were linear with very good correlation coefficients ($r^2>0.9999$) from $0.0125~1{\mu}g/ml$. The limit of detection per sample injection ($20{\mu}l$) was $0.81ng/{\mu}l$ and limit of quantification was $2.47ng/{\mu}l$. The method showed good intra-day precision (%RSD 0.08 ~ 0.27%) and inter-day precision (%RSD 0.32 ~ 1.19%).

Simultaneous Determination of Flavanone Glycosides in the Fruit of Citrus paradisi and C. grandis by HPLC-PDA

  • Piao, Xiang-Lan;Wu, Qian;Han, Saem;Kim, Hyun-Young;Lee, Sang-Hyun
    • Natural Product Sciences
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    • v.17 no.4
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    • pp.337-341
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    • 2011
  • An HPLC (high-performance liquid chromatography)-PDA (photodiode array) detection method was established for the determination of naringin, hesperidin and neohesperidin in the fruit of Citrus paradisi and C. grandis. The flavonoids were separated in less than 20 min using an YMC RP 18 column with isocratic elution using acetonitrile and water (23 : 77, v/v) at a flow rate of 1 ml/min, and a PDA detector. The levels of naringin, hesperidin and neohesperidin were 1345.92, 950.62, and 2078.82 ${\mu}g/g$, respectively, in the peel, and 102.43, 59.13, and 86.68 ${\mu}g/g$, respectively, in the flesh of C. paradisi. In C. grandis, the levels of naringin, hesperidin and neohesperidin were 3530.56, 80.00, and 5.26 ${\mu}g/g$, respectively, in the peel, and 59.59, 7.43, and 38.41 ${\mu}g/g$, respectively, in the flesh. The total content was highest in the peel, reaching 0.44% and 0.36% in C. paradisi and C. grandis, respectively, while the flesh contained only 0.025% and 0.011%, respectively. Therefore, the peels of C. paradisi and C. grandis are necessary for the processing and utilization of flavonoids.

Isolation and Quantitative Analysis of Tricin from Ears of Phragmites communis (갈대이삭으로부터 Tricin의 분리 및 함량 분석)

  • Woo, Hyun Sim;Lee, Seung-Young;Hwang, Buyng Su;Jeong, Sang-Chul;Kim, Dae Wook
    • Korean Journal of Pharmacognosy
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    • v.48 no.1
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    • pp.77-81
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    • 2017
  • The aim of this study was to investigate a validation method for determination of tricin in Phragmites communis ears. Tricin was isolated with chromatographic methods and used as the standard substances for quantitative analysis. The structural determination was characterized by comparing their NMR spectral data with values reported in the literature. For validation, the specificity, linearity, precision, accuracy, detection limits, and quantification limits of tricin was measured by HPLC. The results show that the specificity was satisfied with retention time and diode array detector (DAD) spectrum by analysis of tricin using HPLC. The limits of detection (LOD) for tricin was 0.84 mg/ml. Recovery of tricin was 98.80~108.22% with R.S.D values less than 2%. Intra-day and inter-day precisions of tricin in P. communis ears were 99.96~100.96% and 99.01~100.40%, respectively. Therefore, application of tricin was validated by an analytical method as major compound in P. communis ears.

A study on simultaneous determination of residual sulfonamides in livestock productions by high performance liquid chromatography (HPLC를 이용한 축산식품중 잔류 설폰아미드제의 동시분석법 연구)

  • 황래홍;김영수;윤은선;김기근;이규학
    • Korean Journal of Veterinary Service
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    • v.18 no.3
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    • pp.13-28
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    • 1995
  • This study was carried out to explore the most sensitive and useful method for simultaneous determination of five sulfa drugs(sulfamethazine, sulfamerazine, sulfamonomethoxine, sulfadimethoxine, sulfaquinoxaline) in livestock productions(pork muscle, bovine muscle, chicken muscle, milk ) by HPLC with UV detector and reverse phase column. The results obtained were as follows:1. For mobile phase acetonitrile-0.01M ammonium acetate (23:77) showed more applicable sensitivity and retention times than acetonitrile-1% acetic acid(23:77). Thus acetonitrile-0.01M ammonium acetate(23:77) selected and applied to the modification test, from which it was found pH 6.75 was the most adequate. 2. Optimal wavelength of UV for SMT(sulfamethazine), SMR(sulfamerazine), SMM(sulfamonomethoxine), SD(sulfadimethoxine), and SQ(sulfaquinoxaline) were 266, 266, 265, 269 and 250nm, respectively, and that for simultaneous application it was 263nm. 3. The average recovery rate by extractant(chloroform, dichloromethane, chlorform+dich-loromethane) in the classic method was not significantly different(p>0.05) but that by chloroform higher than the others. Thus chloroform was found to be adequate as extractant in this classic method. 4. The average recovery rate was 86.5% by the MSPD(matrix solid phase disperse) method, which was significantly higher than that by the classic method(p<0.05). Also the recovery rates by method were significantly different(p<0.05) in accordance with sample and type of drug. The MSPD method was much superior to classic method on clean-up.

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Simultaneous Determination of Four Compounds from Artemisia capillaris using High Performance Liquid Chromatography-Ultraviolet Detector (HPLC-UVD) and Their Quantitative Study in Artemisia Genus

  • Ko, Kiwon;Hong, In Kee;Cho, Hyun-Jong;Kim, Young Ho;Yang, Heejung
    • Natural Product Sciences
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    • v.24 no.2
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    • pp.109-114
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    • 2018
  • Artemisia capillaris Thunb. (Compositae) is a native herb of East Asian countries and has used for the treatment of jaundice, high liver fever, and digestive diseases for a long time, as well as being developed as the source of herbal preparations until now. The major components from A. capillaris were chlorogenic acid (1) and its derivatives substituted with caffeoyl moieties, such as 3,5-dicaffeoylquinic acid (2) and 4,5-dicaffeoylquinic acid (3), and coumarins, such as scoparone. In the study, four compounds, chlorogenic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and scoparone (4) in the 70% ethanolic extract of A. capillaris were simultaneously determined by using HPLC-UVD system. This method was validated with the terms of linearity, precious and accuracy according to ICH guidelines. The developed method was successfully applied for the quantitative analysis of Artemisia genus, A. capillaris, A. iwayomogi, A. princeps, and A. argyi, distributed in Korea.

The Assessment of Carbendazim, Cyazofamid, Diethofencarb and Pyrimethanil Residue Levels in P. ginseng (C. A. Meyer) by HPLC

  • Choi, Jeong-Heui;El-Aty, A.M.Abd;Park, Young-Seok;Cho, Soon-Kil;Shim, Jae-Han
    • Bulletin of the Korean Chemical Society
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    • v.28 no.3
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    • pp.369-372
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    • 2007
  • A fast and simple high-performance liquid chromatography (HPLC) method for the simultaneous determination of four pesticides having fungicide properties has been proposed for Panax ginseng, C. A. Meyer grown for 4, 5, or 6 years. Analytical separation was performed on C18 columns using ultraviolet detector under gradient conditions. Spiked blank samples were used as standards to counteract the matrix effect observed in the chromatographic determination. The HPLC response for all pesticides was linear, with determination coefficients > 0.9986. The average rate of recovery for pesticides spiked with 2 fortification levels was > 72% with relative standard deviations < 9%. The limits of quantification (LOQ) ranged from 0.03 to 0.16 ppm. These LOQs were lower than the respective maximum residue limits (MRL) established by the Korean Food and Drug Administration (KFDA), except for cyazofamid. The proposed method was used to determine pesticide residue levels in samples of ginseng obtained from Jeonnam Province (Republic of Korea). None of the pesticides were found in ginseng samples grown for 4, 5, or 6 years.

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

  • Yu, Young-Beob;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.30 no.6
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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.