• Title/Summary/Keyword: Human plasma

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Determination of Eupatilin in Human Plasma by Liquid Chromatography/Electrospray Ionization Tandem Mass Spectrometry

  • Lee, Hye-Won;Ji, Hye-Young;Kim, Hui-Hyun;Kim, Sook-Jin;Kim, Soon-Hoe;Kim, Won-Bae;Lee, Hye-Suk
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.223.2-223.2
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    • 2003
  • A rapid, sensitive and selective liquid chromatography-tandem mass spectrometric (LC/MS/MS) method for the determination of eupatilin in human plasma was developed. Eupatilin and internal standard, (S)-[N-3-(4-(2-(1-methyl-5-tetrazolyl)-pyridine-5-y1)- 3-fluorophenyl)-2-oxo-5-oxazolidinyl]methyl acetamide (DA-7867) were extracted from human plasma by liquid-liquid extraction and analyzed on a phenyl-hexyl column with the mobile phase of acetonitrile-ammonium formate (10 mM, pH 3.0) (60:40, v/v). (omitted)

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Comparison of the Plasma and Urinary Carnitine Profiles between Omnivore and Vegetarian Female College Students

  • Cha, Youn-Soo;Cho, Sang-Woon;Sung, Mi-Kyung
    • Journal of Community Nutrition
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    • v.4 no.2
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    • pp.78-82
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    • 2002
  • This study compared the effect of Korean vegetarian and omnivorous diets on plasma carnitine concentrations and urinary carnitine excretion. Twenty lactoovovegetarian and twenty omnivorous female college students consented to participate in this study. Daily nutritional intake and plasma and urinary nonesterified carnitine (NEC), acid-soluble acylcarnitine (ASAC), acid-insoluble acylcarnitine (AIAC), and total carnitine (TCNE) were determined. Daily protein, fat, retinol, vitamin B$_2$and vitamin B$\_$12/ intakes were significantly lower for vegetarians, however, fiber, carbohydrate, $\beta$-carotene, folic acid and vitamin C consumptions were much higher for vegetarians than omnivores. There were no differences in plasma NEC, ASAC, AIAC and TCNE concentrations between the two groups. Urinary carnitine excretion was lower in vegetarians, but only the differences in ASAC and TCNE excretions were statistically significant. These results suggest that the lower excretion of ASAC in vegetarians may be a reflection of their lipid metabolic state and that Korean vegetarian diets may accommodate lower carnitine intakes through efficient urinary conservation of carnitine.

Validation of Kinetic Method for the PKA Assay in Plasma-Derived Products

  • Shin, In-Soo;Hong, Choong-Man;Koh, Hyun-Chul;Hong, Seung-Hwa
    • Biomolecules & Therapeutics
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    • v.13 no.1
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    • pp.59-63
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    • 2005
  • A kinetic assay was carried out in order to compare the ability of detection for prekallikrein activator(PKA) in plasma-derived products with that of an endpoint assay and a commercial method. Using these methods, 9 human albumin preparations were assayed and compared to each other. The coefficient of variation between the Kinetic assay and the end point assay was found within 6.6% and this result showed that two methods were highly correlative and the end point assay could act as a replacement of the kinetic assay. Another important goal of this study was to investigate the reproducibility among laboratories on the kinetic assay. A collaborative study was performed to validate the kinetic method with intra and inter assays. The coefficient of variation for the intra assay of each laboratory was less than 4% and that for between individuals in the inter assay was 4.1%. These results revealed that the kinetic assay showed good reproducibility. The contents of PKA in plasma-derived products were also determined by the kinetic assay. As a result, it was found that trace amounts of PKA were present in 32 human immunoglobulin preparations, however the average concentration of PKA in 171 albumin preparations was 5.8 IU/mL.

Determination of Clotiazepam in the Plasma Using Gas Chromatography/Mass Spectrometry with an Ion-Trap Detector and its Application to Pharmacokinetics in Healthy Volunteers

  • Kwon, Oh-Seung;Kim, Seung-Yong;Chung, Youn-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.2
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    • pp.123-129
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    • 2006
  • A method determining the plasma concentration of clotiazepam was developed by using gas chromatography/mass spectrometry with an ion-trap detector and was validated for applying pharmacokinetics to human volunteers orally taken 5 mg dose of clotiazepam. The detection limit was 1 ng/ml and the limit of quantitation was 5 ng/mt. Intraday reproducibility and accuracy bias % were less than 8.2 and 10.2% with inter-day variations for those being within 7.0 and 13.8%, respectively. The recovery of clotiazepam was higher than 87%. The principal pharmacokinetic parameters were determined from the plasma concentration-time plot by non-compartmental or two-compartmental analysis. In non-compartmental analysis, the elimination half-life of 10.4 hr and the area under the curve of 651.3 ng hr/ml were determined, and the maximal concentration (158.6 ng/ml) in the plasma was obtained at 0.56 hr post-dose. The developed method can be appropriate to apply pharmacokinetics and bioequivalence of clotiazepam.

Capillary Size-exclusion Chromatography as a Gel-free Strategy in Plasma Proteomics

  • Cho, Man-Ho;Wishnok, John S.;Tannenbaum, Steven R.
    • Molecular & Cellular Toxicology
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    • v.1 no.2
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    • pp.87-91
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    • 2005
  • Although 2D-PAGE has been widely used as the primary method for protein separation, difficulties in displaying proteins with an extreme values of isoelectric paint (pI), molecular size and hydrophobicity limit the technique. In addition, time consuming steps involving protein transfer and extraction from the gel-pieces can result in sample loss. Here, we describe a novel protein separation technique with capillary size-exclusion chromatography (CSEC) for rapid protein identification from human plasma. The method includes protein fractionation along with molecular size followed by in-solution tryptic digestion and peptide analysis through reversed phase liquid chromatography (RPLC) coupled to nanoflow electrospray-tandem mass spectrometry (ESI-MS/MS). Tryptic peptides are applied an a $100\;{\mu}m\;i.d.{\times}10mm$ length pre-column and then separated on a $75\;{\mu}m{\times}200mm$ analytical column at -100 nL/min flaw rate. Proteins were identified over the wide ranges of pI (3.7-12.3) when this technique was applied to the analysis of $1-2\;{\mu}L$ of human plasma. This gel-free system provides fast fractionation and may be considered a complementary technique to SDS-PAGE in proteomics.

Pharmacokinetics rind Tissue Distribution of a Recombinant truman Erythropoietin, GC-rhEPO (유전자 재조합 사람형 erythropoietin, GC-rhEPO의 약물동태 및 조직분포)

  • 김선돈;한성규;이호성;김성남;정원휘;백대현;조은성;허재욱;류판동
    • Biomolecules & Therapeutics
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    • v.8 no.2
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    • pp.171-178
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    • 2000
  • To evaluate the pharmacokinetic properties and tissue distribution of a newly developed recombinant human erythropoietin (GC-rhEPO), we analyzed the plasma and tissue levels of erythropoietin by an ELISA after intravenous (IV) and subcutaneous (SC) adminstration to the male rats at the doses of 20, 100, 500 or 2,500 unit/kg. After single IV bolus injection of GC-rhEPO, the plasma concentration was rapidly increased and decreased with two phases with half-lives of 13.4 min and 2.94 hours. AUC was increased dose- dependently but plasma half-lives remained constant regardless of GC-rhEPO doses. Following SC administration, the plasma concentration increased slowly with half-life of 9.2 hours and reached peak at 8 hours. Mean residence time and bioavailability were 18.2 hours and 44%, respectively. After single IV dose of 100 unit/kg, tissue GC-rhEPO level was higher in bone marrow and spleen, while the depletion rate was slower in liver and bone marrow, indicating the higher affinity of GC-rhEPO to bone marrow. Taken together, the experimental results indicate that GC-rhEPO contained the typical pharmacokinetic properties and the tissue distribution patterns inherent to human erythropoietin.

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Development of Nonthermal Bioplasma Source Applicable to Human Liquid Fluids

  • Min, Boo-Ki;Oh, Hyun-Joo;Song, Ki-Baek;Uhm, Han-Sup;Kang, Seung-Oun;Choi, Eun-Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.08a
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    • pp.327-327
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    • 2011
  • A nonthermal bioplasma source was developed for application to human liquid fluids by making use of nano-size tungsten tips. Characteristics of the plasma source are investigated. Here we have used the AC voltage system. The bioplasma source generated by a tungsten tip with quartz tube and ground electrode is a low-temperature plasma without making any noticeable damage to cells at a low power operation. The breakdown voltage and current signals are measured by high voltage probe (Tektronix P6015A) and current probe (P6021). Variation of breakdown temperature near the tip electrode is larger than that in the neighborhood of ground electrode. Bubble formation during discharge has been recorded and investigated by using the high speed camera. The existence and behavior of hydroxyl and superoxide radicals are detected and measured by spectrometers. The electrical and optical properties of breakdown characteristics are also investigated.

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Bioavailability of Cefaclor Capsules Using an Improved Analytical Method of Cefaclor in Human Plasma (개선된 사람 혈장중 세파클러 농도 정량법을 이용한 세파클러 캡슐의 생체이용률 측정)

  • Kim, Tae-Wan;Song, Ok-Kyoung;Han, Sun-Young;Cao, Qing-Ri;Park, Mi-Jin;Kang, Sung-Ha;Shin, Kwan-Seog;Cui, Jing-Hao;Lee, Beom-Jin
    • Journal of Pharmaceutical Investigation
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    • v.35 no.2
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    • pp.117-122
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    • 2005
  • After establishing improved HPLC analytical method of cefaclor in human plasma samples, a bioavailability study of cefaclor capsules was conducted according to the guidelines of Korea Food and Drug Administration (KFDA). The standard calibration curve using an HPLC with UV detector was constructed in a range of $0.0324{\sim}16\;{\mu}g/ml$. The 6% perchloric acid instead of 6% trichloroacetic acid was used to precipitate plasma protein. The HPLC chromatograms were precisely and accurately resolved when spiked with human plasma spiked with cefaclor and cephalexin (internal standard). Twenty healthy male Korean volunteers received two commercial cefaclor capsules, $Neocef^{\circledR}$ capsule (Jinyang Pharm. Co., Ltd) or $Ceclor^{\circledR}$ capsule (Lilly Korea. Co., Ltd.) at the 250 mg cefaclor in a $2{\times}2$ crossover study. There was a one-week washout period between the doses. Plasma concentrations of cefaclor were monitored for 8 hours after oral drug administration. $AUC_t$ the area under the plasma concentration-time curve from time zero to 8 hr (13 points), was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_t$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the cross-over design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for $Neocef^{\circledR}/Ceclor^{\circledR}$ were $0.9049{\leq}{\delta}{\leq}1.226$, respectively. These values were within the acceptable bioequivalence intervals of 0.80-1.25. Thus, our study demonstrated the bioequivalence of $Neocef^{\circledR}/Ceclor^{\circledR}$ with respect to the extent of absorption.

Evaluation of Pasma Folate and Total Homocysteine in Korean Alcoholics

  • Hyesun Min;Kim, Chang-Su;Jungsook Seo
    • Journal of Community Nutrition
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    • v.1 no.1
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    • pp.60-65
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    • 1999
  • Chronic alcoholism often leads to folate deficiency. In recent years it has been reported that mild elevation of plasma homocysteine (Hcy) is associated with an increased risk of coronary artery disease. In the present study we investigated the effects of chronic ethanol consumption on folate status and the relation between plasma folate and Hcy. A human study was conducted to determine plasma folate, total Hcy, cysteine(Cys), total cholesterol and hemoglobin(Hb) concentrations in 44 Korean alcoholics(men aged 30 to 50yr) and 45 Korean non-alcoholic subjects(men aged 30 to 50 yr). In alcoholic subjects, 52.6% were folate deficient and 34.2% were marginally deficient, which suggested that most alcoholics were subnormal in folate status. Plasma total Hcy concentration of alcoholics was twice as high as in control subjects (p<0.001). We found a negative correlation between plasma folate and plasma total Hcy(r=-0.271, p<0.05) and a positive correlation between plasma folate and plasma Cys(r=0.249, p<0.05) in total subjects. Hb concentrations in alcoholics was significantly lower than in control subjects, but there was no difference in total cholesterol concentration between alcoholics and controls. These results suggest that chronic alcohol consumption may impair the disposal of Hcy by interfering with folate metabolism.

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Plasma Aminogram and Urinary Excretion of Free Amino Acids in Adult Vegetarians Compared with AGe-Matched Omnivores in Korea

  • Park, Tae-Sun;Chang, Jun-Sung;Sung, Mi-Kyung
    • Preventive Nutrition and Food Science
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    • v.3 no.4
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    • pp.368-373
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    • 1998
  • Plasma amino acid concentration and Urinary exretion of free amino acids were measured in health female vegetarians(n=20, 19.9 $\pm$0.43 years old ) and age-mateched imnivores(n=20, 21.9$\pm$0.38years old) in Korean. differences infasting plasma amino acid concentrations and plasma aminogram pattern were not spectacular between the vegetarian and omnivore controls. Compared to the omnivores, vegetarians showed significantly lower plasma levels of methionine , phenylalanine, $\alpha$-aminobutyrate, citrulline, phosposerine and tarurine, and significantly higher plasma concentrations of arginine, $\alpha$-aminobutyrate, cirtrulline, phosphosierine and taurine, and significantly higher plasma concentrations of arginine, $\alpha$-aminoadipate, asparagine, aspartate, glutamate and ornithine. Although these differences were statistically significant, they were all within the normal range (21~70% differences )for human adults. Most of the urinary amino acids (nmol/mg creatinine or $\mu$mol/24 hr urine) were excreted to significantly lesser degree in vegetarians than was the case in omnivore controls. For almost every individual free amino acid, plasma concentration did not significantly correlate with urinary excretion level. These results indicate that vegetarians excreted less amino acids in their urine than did dominivores, most probably in an effort to maintain amino acid homeostasis to an altered dietary protein intake level and/or amino acid composition of their diets.

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