• Title/Summary/Keyword: Human plasma

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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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A Sensitive and Rapid LC-MS/MS Method for Determination of Berberine in Human Plasma

  • Kim, Jae Hee;Mai, Xuan-Lan;Kim, Kyu Yeon;Sim, Min-Seok;Lee, So-Yeon;Seo, Hee-Won;Lee, Gunhee;Kim, Dae-Jung;Kim, Kyeong Ho
    • Mass Spectrometry Letters
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    • v.10 no.2
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    • pp.56-60
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    • 2019
  • Coptidis Rhizoma (CR) has been used widely in traditional medicine to treat common diseases. This study aimed to develop a high-sensitivity liquid chromatography-tandem mass (LC-MS) spectrometry method for the evaluation of the pharmacokinetics of a new natural product that contain CR extract with the main bioactive compound, berberine, at trace concentrations. Human plasma samples were pretreated with methanol by a protein precipitation method. Berberine was analyzed on a Kinetex C18 column ($2.1mm{\times}50mm$, $100{\AA}$, $1.7{\mu}m$) using a mobile phase of 10 mM ammonium formate/0.1% formic acid in water (A) and acetonitrile (B) (50:50, v/v) with a flow rate of 0.25 mL/min. The analyte was detected by using electrospray ionization in positive mode with multiple reaction monitoring (MRM). The method was sensitive, with a lower limit of quantification of 1 pg/mL, which has not been previously obtained. The method was validated (over the range of 1-50 pg/mL) and applied successfully for the pharmacokinetic study of human plasma samples.

Cold Ethanol Fractionation and Heat Inactivation of Hepatitis A Virus During Manufacture of Albumin from Human Plasma

  • Kim, In-Seop;Park, Yong-Woon;Lee, Sung-Rae;Sung, Hark-Mo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.1
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    • pp.65-68
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    • 2004
  • The purpose of the present study was to examine the efficacy and mechanism of fraction IV cold ethanol fractionation and pasteurization (60$^{\circ}C$ heat treatment for 10 h), involved in the manufacture of albumin from human plasma, in the removal and/or inactivation of the hepatitis A virus (HAV). Samples from the relevant stages of the production process were spiked with HAV and the amount of virus in each fraction then quantified using a 50% tissue culture infectious dose (TCID$\_$50/). HAV was effectively partitioned from albumin during the fraction IV cold ethanol fractionation with a log reduction factor of 3.43. Pasteurization was also found to be a robust and effective step in inactivating HAV, where the titers were reduced from an initial titer of 7.60 log TCID$\_$50/ to undetectable levels within 5 h of treatment. The log reduction factor achieved during pasteurization was $\geq$4.76. Therefore, the current results indicate that the production process for albumin has sufficient HAV reducing capacity to achieve a high margin of virus safety.

Simplified HPLC Method for the Determination of Mirtazapine in Human Plasma and Its Application to Single-dose Pharmacokinetics

  • Gwak Hye-Sun;Lee Na-Young;Chun In-Koo
    • Biomolecules & Therapeutics
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    • v.14 no.1
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    • pp.40-44
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    • 2006
  • Mirtazapine is an antidepressant agent with dual action on both the noradrenergic and serotonergic neurotransmitter systems. A simple high performance liquid chromatographic method has been developed and validated for the quantitative determination of mirtazapine in human plasma. A reversed-phase Cl8 column was used for the determination of mirtazapine with a mobile phase composed of 0.01M ammonium acetate solution (pH 4.2) and acetonitrile (75:25, v/v%) at a flow rate of 1.2 mL/min. Terazosin hydrochloride was used as an internal standard. The fluorescence detector was set at excitation and emission wavelengths of 290 and 350 nm, respectively. Intra- and inter-day precision and accuracy were acceptable for all quality control samples including the lower limit of quantification of 3 ng/mL. Mirtazapine was stable in human plasma under various storage conditions. This method was used successfully for a pharmacokinetic study using plasma samples after oral administration of a single 30 mg dose as mirtazapine base to 8 healthy volunteers. The maximum plasma concentration of mirtazapine was $64.1{\pm}28.0ng/mL$ at 1.8 h, and the area under the curve and elimination half-life were calculated to be $674.1{\pm}218.5ng\;h/mL\;and\;23.4{\pm}3.8h$, respectively.

Assay of Midazolam in Human Plasma by Gas-Liquid Chromatography with Nitrogen-Phosphorus Detection

  • 신호상;홍춘표;Yun-Suk Oh-Shin;강보경;이경옥;이규범
    • Bulletin of the Korean Chemical Society
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    • v.19 no.5
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    • pp.524-526
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    • 1998
  • A sensitive and specific method is described for the determination of midazolam in human plasma. The drug was extracted from 1 mL of carbonate buffered plasma (pH 9.6) with 8 mL of diethyl ether. Famprofazone was used as internal standard. The organic phase was evaporated to dryness. The residue was dissolved in methanol for the direct analysis by gas chromatograph-nitrogen phosphorus detector system. In the concentration range of 1-5000 ng/mL, the calibration curve was linear. The coefficients of variation from the precision test were <6% at the range of the concentration of 0.10-2.00 μg/mL and the detection limit for midazolam in 1 mL of plasma was o.5 ng. This assay is more sensitive, selective, simple and rapid than earlier methods. Plasma midazolam concentrations were determined by this method after administration of midazolam.

Partitioning and Inactivation of Viruses by Cold Ethanol Fractionation and Pasteurization during Manufacture of Albumin from Human Plasma

  • Kim, In-Seop;Eo, Ho-Gueon;Chang, Chon-Geun;Lee, Soung-Min
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.858-864
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    • 2000
  • The purpose of the present study was to examine the efficacy and mechanism of the fraction IV cold ethanol fractionation and pasteurization ($60^{\circ}C$ heat treatment for 10h) steps, involved in the manufacture of albumin from human plasma, in the removal and/or inactivation of blood-born viruses. A variety of experimental model viruses for human pathogenic viruses, including the Bovine viral diarrhoea virus (BVDV), Bovine herpes virus (BHV), Murine encephalomyocarditis virus (EMCV), and Porcine parvovirus (PPV), were selected for this study. Samples from the relevant stages of the production process were spiked with the viruses, and the amount of virus in each fraction was then quantified using a 50% tissue culture infectious dose ($TCID_{50}$). The mechanism of reduction for the enveloped viruses (BHV and BVDV) during fraction IV fractionation was inactivation rather than partitioning, however, it was partitioning in the case of the non-enveloped viruses (EMCV and PPV). The log reduction factors achieved during fraction IV fractionation were ${\geq}6.9$ BHV, $\geq5.2$ for BBDV, 4.9 for EMC, and 4.0 for PPV. Pasteurization was found to be a robust and effective step in inactivating the enveloped viruses as well as EMCV. The log reduction factors achieved during pasteurization were $\geq7.0$ for BHV, $\geq6.1$ for BVDV, $\geq6.3$ for EMCV, and 1.7 for PPV. These results indicate that the production process for albumin has sufficient virus-reducing capacity to achieve a high margin for virus safety.

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Rapid Quantitative Analysis of Vancomycin in Human Plasma and Urine Using LC-MS/MS (LC - MS/MS를 이용한 혈장과 뇨중에서 Vancomycin의 빠른정량분석)

  • Kim, Hohyun;Roh, Hyeongjin;Han, Sang-Beom
    • Analytical Science and Technology
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    • v.15 no.5
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    • pp.410-416
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    • 2002
  • In this study, a new quantitative analytical method has been developed for the rapid determination of vancomycin in human plasma and urine using liquid chromatography/tandem mass spectrometry (LC - MS/MS). Chromatography was carried out on a $C_{18}$ XTerra MS column ($2.1{\times}30mm$) with a particle size of $3.5{\mu}m$. The mobile phase was 0.25% formic acid in 10% acetonitrile and the flow rate was $250{\mu}L/min$. Vancomycin and caffeine (internal standard) were detected by MS/MS using multiple reaction monitoring (MRM). Vancomycin gives a predominant doubly protonated precursor molecule ($[M+2H]^{2+}$) at m/z 725.0 and a corresponding product ion of m/z 100.0. Detection of vancomycin was good, accurate and precise, with a limit of detection of 1 nM in plasma. The calibration curves for vancomycin in human plasma was linear in a concentration range of $0.01{\mu}M$ - $100{\mu}M$ for plasma. This method has been successfully applied to determine the concentration of vancomycin in human plasma and urine from pharmacokinetic study and relative studies.

Effects of Human Plasma Lipid Transfer Protein on the Distribution of Lipids Between Lipoprotein Fractions of Rat Plasma (인체의 혈장에서 분리한 지질전이단백질이 흰쥐의 혈장 Lipoprotein 의 지질분포에 미치는 영향)

  • 최영선
    • Journal of Nutrition and Health
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    • v.19 no.5
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    • pp.296-303
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    • 1986
  • Rat lipoprotein-deficient plasma possessed a lipid tramsfer inhibitory activity when it was added to purified human plasma lipid transfer protien, while it lacked a lipid transfer activity. Incubation of whole rat plasma with partially purified human lipid transfer protein resulted in big changes in lipid distribution of rat plasma lipoproteins. There w was a 4-fold increase in cholesteryl ester(CE) and 4 47 % reduction in triglyceride(TG) in very low density lipoproteins after 2싹lour incubation. In high density lipoprotein $2(HDL_2)$ there was a 9­fold increase in TG and 33 % reduction in CEo HDL3 had 82 % reduction in CE. The result indi­c cates that the absence of the lipid transfer activity in rat plasma can be ascribed not to the inability of rat lipoproteins to serve as substrates but to the lack of 야Ie lipid transfer protein in rat plasma. Th­e erefore, species differences in lipid transfer betwe­e en lipoproteins should be taken into consideration to interpret results of studies on lipoprotein m.eta­b bolism using rats.

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MTHFR Polymorphism and Folate Status of Korean Women of Childbearing Age

  • Choi, Jeong-Hwa;Kim, Hee-Ah;Lim, Hyeon-Sook
    • Nutritional Sciences
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    • v.9 no.1
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    • pp.35-41
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    • 2006
  • It should be concerned to the women with mutated genotype of methylenetetrahydrofolate reductase (MTHFR), C677T or A1298C, since they need more folate than those with wild genotypes. In this study, we evaluated the folate status of Korean women of childbearing age according to their MTHFR polymorpiysm. Dietary folate intakes, plasma and erythrocyte folate concentrations, plasma homocysteine concentrations, and urinary excretions of para-aminobenzoylglutamate (pABG) and para-acetoamidobenzoylglutamate (ApABG) of twenty-five subjects aged between 19 and 35 years old were determined Folate intakes seemed to be inadequate, being only three-quarters of the Korean RDA of folate. More than one-quarter of the subjects was exposed to folate deficiency risk as determined by erythrocyte folate concentration and almost one-quarter of the subjects showed hyperhomocysteinemia, although they had normal plasma folate concentrations. Urinary excretions of pABG and ApABG seemed to be low and ApABG constituted more than $85\%$ of total folate catabolites. There were no significant differences in dietary folate intakes, plasma concentrations of folate and homocysteine, and urinary excretions of pABG and ApABG among the geneotypes of both C677T and A1298C. However, the subjects with 1298AC genotype had significantly lower erythrocyte folate concentration than those with 1298AA. Erythrocyte folate concentration showed an inverse relationship with plasma homocysteine concentration and positive relationships with urinary excretions of pABG and ApABG. The results of this study imply that mutations of 677C$\rightarrow$T and 1298A$\rightarrow$C in the study were not associated with decreased plasma folate and raised plasma homocysteine concentrations. A1298C polymorphism night be, however, more influential on erythrocyte folate concentration than C677T polymorphism, and urinary excretions of folate catabolites, pABG and ApABG, might be reliable indexes of folate nutritional status like plasma homocysteine concentrations.

Advanced Method for Determination of Omeprazole in Plasma by HPLC

  • Kang, Won-Ku;Kim, Dong-Sup;Kwon, Dwang-Il
    • Archives of Pharmacal Research
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    • v.22 no.1
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    • pp.86-88
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    • 1999
  • An advance d and sensitive high-performance liquid chromatographic (HPLC) method for determination of omeparzole in human plasma has been developed. After omeprazole was extracted from plasma with diethylether, the organic phase was transferred to another tube and trapped back with 0.1 N NaOH solution. The alkaline aqueous layer was injected into a reversed-phase C8 column. Lansoprazole was used as an internal standard. The mobile phase consisted of 30% of acetonitrile and 70% of 0.2 M $ KH_{2}PO_{4}$, pH 7.0. Recoveries of the analytes and internal standard were >75.48%. The coefficients of variation of intra- and inter-day assay were <5.78 and 4.59% for plasma samples. The detection limit in plasma was 2 ng/ml. The clinical applicability of this assay method was evaluated by determining plasma concentration-time courses of the respective analytes in 24 healthy volunteers after oral administration 40 mg of omeprazole. The present assay is considered to be simple, accurate, economical and suitable for the study of the kinetic disposition of omeprazole in the body.

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