• Title/Summary/Keyword: High performance Liquid Chromatography

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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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    • v.16 no.5
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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.

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%.

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.

Pharmacokinetics of KR-30075, A Potent Phosphodiesterase III Inhibitor in Rats (포스포디에스테라제 III의 저해물인 KR-30075의 흰쥐에서의 약물속도론)

  • Lee, Kwang-Pyo;Kim, Hyo-Jin;Kwon, Kwang-Il;Cho, Song-Ja
    • YAKHAK HOEJI
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    • v.36 no.3
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    • pp.259-268
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    • 1992
  • A procedure for the determination of KR-30075 and its metabolites in plasma and urine by high performance liquid chromatography is described. For the study of pharmacokinetic properties of KR-30075, a new PDE III inhibitor, the plasma concentration and urinary excretion after an oral administration of KR-30075 (4 mg/kg) in the male rat (Sprague Dawley) were determined by high performance liquid chromatography. The best extraction efficiency of KR-30075 and KR-30072 is obtained with ethyl ether adjusted to pH 4.0. Retention times of both KR-30072 and KR-30075 were within 5 min and resolution was complete at the flow rate of 1.0 ml/min. The sensitivity and specificity of this HPLC assay appears to be satisfactory for the pharmacokinetic study of KR-30075 and its metabolites. One-compartment open model with first-order absorption was applied to evaluate the pharmacokinetic parameters of KR-30075 according to Minimum AIC Estimation. $T_{max}$ was 1 hr, $C_{max}$ was $0.789{\pm}0.31\;{\mu}g/ml$ and elimination half $T_{1/2}$ was 6.31 min after oral administration of 4 mg/kg KR-30075 to male rats.

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Simultaneous Quantitation of Nine Constituents of Fraxinus rhynchophylla using High Performance Liquid Chromatography - Diode Array Detector

  • Ahn, Jong Hoon;Hwang, Bang Yeon;Lee, Mi Kyeong
    • Natural Product Sciences
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    • v.19 no.3
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    • pp.236-241
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    • 2013
  • A high-performance liquid chromatography-diode array detector (HPLC-DAD) method was established for quantitative evaluation of nine constituents of Fraxinus rhynchophylla such as four coumarins, esculin (1), fraxin (2), esculetin (3), fraxetin (4), three lignans, syringaresinol 4,4'-O-${\beta}$-diglucoside (5), pinoresinol 4-O-${\beta}$-glucoside (6), pinoresinol (9), one secoiridoid, oleuropein (7), and one coumarinolignan, cleomiscosin C (8). The preferred chromatographic condition was obtained on Phenomenex Gemini-NX (3 ${\mu}m$, C18 110A, $150{\times}4.60$ mm) and the mobile phase was composed of water and acetonitrile using a gradient elution. The wavelength was set at 220 nm. Extraction condition of these constituents in F. rhynchophylla was also optimized through extraction time, extraction solvent and extraction method using established method. From this study, extraction at $70^{\circ}C$ with the mixture of ethanol and water for more than 12 h was suggested to be good extraction condition for these constituents. Quantitation of nine constituents in different F. rhynchophylla samples was also successfully accomplished with the newly established method.

Monitoring of Biotin Content in Frequently Consumed Foods in Korea (국내 다소비 식품의 biotin 함량 모니터링)

  • Kwon, Jihyun;Cheon, Wonyoung;Lee, Sang-Hoon;Choi, Youngmin;Kim, Younghwa
    • Journal of the Korean Society of Food Culture
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    • v.35 no.3
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    • pp.278-284
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    • 2020
  • In this study, biotin (vitamin B7) contents of frequently consumed foods in Korea were determined by using immunoaffinity column in conjunction with high-performance liquid chromatography (HPLC). The biotin contents of 24 foods of plant origin and 27 foods of animal origin were selected. The highest biotin contents in frequently consumed foods of plant origin were found in red beans (Huinguseul; 11.475 ㎍/100 g). On the other hand, biotin was not detected in any varieties of sorghum. For frequently consumed foods of animal origin, salted pollack roe (7.486 ㎍/100 g) showed the highest biotin content. However, beef and fish contained less biotin. All biotin analyses were conducted under analytical quality control. The limits of detection and limits of quantification of biotin were 0.007 and 0.023 ㎍/100 g, respectively, and the accuracy/recovery percentage was 95.35-105.02%. The precision values were 4.041% (repeatability) and 3.835% (reproducibility). Taken together, our data provide reliable data on the biotin contents of frequently consumed foods in Korea.

Determination of Aflatoxins Using High-Performance Liquid Chromatography with Optimized Fluorescence Detection (HPLC에 의한 aflatoxin 분석법에 관한 연구-형광검출의 최적조건)

  • 김종규
    • Journal of Food Hygiene and Safety
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    • v.13 no.1
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    • pp.41-46
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    • 1998
  • A postcolumn derivatization method was tried for the simultaneous determination of four major aflatoxins ($B_1,\;B_2,\;G_1,\;and\;G_2$) by high-performance liquid chromatography with fluorescence detection. As compared with a previous precolumn derivatization method, it was found that the postcolumn derivatization combined with an electrochemical cell (Kobra cell) was less time-consuming, safer, improved the sensitivity and selectivity, and provided good recoveries for aflatoxin $B_1$ (88.9%) and $G_1$ (100.5%). This method showed linearity from 10~100 ppb for aflatoxin $B_1\;and\;G_1$, and from 3~30 ppb for aflatoxin $B_2\;and\;G_2$. However, aflatoxin Bz and Gz were not detected satisfactorily although they showed good resolution.

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Induction of Salicylic Acid Production in Pepper by Yeast Cell Wall Extract (효모세포벽추출물에 의한 고추내 salicylic acid 생성유도)

  • Kang, Dae Sun;Cho, Soo Muk;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.299-302
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    • 2012
  • Yeast cell wall extract (YCWE) was treated on leaves and roots of pepper seedlings at the dosage of 4 mg/mL and salicylic acid (SA) production in pepper was detected by ultra high performance liquid chromatography (UHPLC). The SA production in pepper stem was induced by YCWE. SA was produced at the highest level of 20.29 ${\mu}g/g$ after 48 hrs of foliar spray with YCWE, which is 3.7 times higher than that of root perfusion with YCWM. SA production was gradually reduced after 72 hrs of YCWE treatment.

Determination of ${\gamma}-Aminobutyric$ Acid and Glutamic Acid in Rat Brain by High Performance Liquid Chromatography with Electrochemical Detection (HPLC-ECD를 이용한 흰쥐의 뇌 중 감마 아미노부티르산 및 글루탐산의 정량)

  • 강종성;이순철
    • YAKHAK HOEJI
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    • v.43 no.3
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    • pp.300-305
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    • 1999
  • A sensitive and efficient assay method was applied to determine the level of glutamic acid (GA) and ${\gamma}-aminobutyric$ acid (GABA) in frontal cortex and hippocampus of rat administrated with ethanol and drugs. The compounds were derivatized with ο-phthalaldehyde (OPA) and 2-mercaptoethanof for precolunm analysis. The condition for the simultaneous determination of GA, GABA and $\beta-aminobutyric$ acid (BABA) by high performance liquid chromatography with electrochemical detection was reverse phase $C_{18}$ column as stationary phase, 0.1 M phosphate buffer containing 0.1 mM $Na_4EDTA$ : methanol = 55:45 (v+v) pH 3.8 as mobile phase and 0.7V electrode voltage. The stability of reaction product of GA, GABA and BABA with OPA could be increased by adding the same volume of polyethylene glycol 400 to reaction mixture. The GABA level in frotal cortex of rat was significantly decreased by the administration of picrotoxin and diazepam, but it was significantly increased by the administration of red ginseng total saponin, N-methyl-D-glucamine and (-)-deprenyl.

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Incidence and Level of Aflatoxins Contamination in Medicinal Plants in Korea

  • Lee, Sung Deuk;Yu, In Sil;Jung, Kweon;Kim, Yeon Sun
    • Mycobiology
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    • v.42 no.4
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    • pp.339-345
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    • 2014
  • During 2011~2013, a total of 729 samples for 19 types of medicinal plant were collected from Seoulyekryungsi in Seoul, Korea, and investigated for the presence of aflatoxins. The samples were analyzed using immunoaffinity column cleanup and high-performance liquid chromatography coupled to a fluorescence detector after post-column derivatization. Aflatoxins were found in 124 out of the 729 analyzed samples: 65 containing aflatoxin B1 (AFB1), 24 with aflatoxin B2 (AFB2), 15 with aflatoxin G1 (AFG1), and 20 samples with aflatoxin G2 (AFG2). The ranges for positive samples were $0.1{\sim}404.7{\mu}g/kg$ for AFB1, $0.1{\sim}10.0{\mu}g/kg$ for AFB2, $0.1{\sim}635.3{\mu}g/kg$ for AFG1, $0.1{\sim}182.5{\mu}g/kg$ for AFG2, and $0.1{\sim}1,043.9{\mu}g/kg$ for total aflatoxins. Most of the medicinal plant samples (721, 98.9%) were below legal limits, but 8 samples exceeded the legal limits of 10 and $15{\mu}g/kg$ established by the Korean standard for AFB1 and total aflatoxins (the sum of AFB1, AFB2, AFG1 and AFG2), respectively.