Detection of hemoglobin-based oxygen carriers in human serum for doping analysis: confirmation by size-exclusion HPLC.

Abstract:

BACKGROUND:Hemoglobin-based oxygen carriers (HBOCs) are being developed as potential substitutes for the oxygen-carrying functions of erythrocytes, but athletes may obtain and experiment with HBOCs as an illicit means of enhancing oxygen transport. An electrophoretic technique has been developed to screen for the presence of HBOCs in blood samples (Lasne et al. Clin Chem 2004;50:410-5). Interest has focused on complementary methods that can provide legally defensible scientific evidence for the presence of HBOCs in blood samples collected for doping control. METHODS:The aim of this research was to develop a size-exclusion SEC-HPLC technique to identify in plasma or serum samples the presence of HBOCs that are currently under development. This method was also used to detect a polymerized bovine hemoglobin (Hemopure) after infusion in 12 healthy males. RESULTS:The chromatograms of all HBOCs tested were clearly separated from the 54-min peak associated with human hemoglobin dimers. It was possible to differentiate between the different HBOC products based solely on their chromatographic profiles, provided they were at high concentrations. Differences were discernible not only based on the presence (or absence) of peaks, but also the separation between respective peaks. The profiles for serum samples collected from the men immediately after infusion of Hemopure showed a distinctive profile. The shape of the chromatographic profile remained consistent for at least 48 h. CONCLUSIONS:Under the analytical conditions reported here, SEC-HPLC was able to separate native hemoglobin from the modified hemoglobin molecules present in each of the HBOC products studied. In tandem with electrophoretic screening, SEC-HPLC provides evidence of the presence of HBOCs and can therefore be regarded as a method that satisfies the criteria for use in an antidoping control setting.

journal_name

Clin Chem

journal_title

Clinical chemistry

authors

Varlet-Marie E,Ashenden M,Lasne F,Sicart MT,Marion B,de Ceaurriz J,Audran M

doi

10.1373/clinchem.2003.026591

subject

Has Abstract

pub_date

2004-04-01 00:00:00

pages

723-31

issue

4

eissn

0009-9147

issn

1530-8561

pii

clinchem.2003.026591

journal_volume

50

pub_type

杂志文章
  • High urinary concentrations of activin A in asphyxiated full-term newborns with moderate or severe hypoxic ischemic encephalopathy.

    abstract:BACKGROUND:Hypoxic ischemic encephalopathy (HIE) is a major cause of permanent neurological disabilities in full-term newborns. We measured activin A in urine collected immediately after birth in asphyxiated full-term newborns, and assessed the ability of the measurements to predict the occurrence of perinatal encephal...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2005.062604

    authors: Florio P,Luisi S,Moataza B,Torricelli M,Iman I,Hala M,Hanna A,Petraglia F,Gazzolo D

    更新日期:2007-03-01 00:00:00

  • Clinical applications of protein determinations in biological fluids other than blood.

    abstract::Over the past decade, significant improvements in immunochemical and electrophoretic techniques have enabled collection of heretofore unavailable data on proteins in biological fluids, greatly increasing our understanding of protein physiology in the various body compartments and providing the foundation for clinical ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Killingsworth LM

    更新日期:1982-05-01 00:00:00

  • Low bias in assayed values of lipoprotein antigens--lipoprotein(a) and apolipoproteins A-I and B--in midday postprandial blood specimens compared with morning fasting specimens.

    abstract::Two-hour postprandial specimens have a -14% proportional bias for lipoprotein(a) [Lp(a)], a -0.035 g/L systematic bias for apolipoprotein (apo) A-I, and a -9% proportional bias for apo B, compared with values in 12-h fasting specimens. Although a physiological hemodilution appears to account for a portion of these bia...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Emancipator K

    更新日期:1992-03-01 00:00:00

  • Leukocyte counts in cerebrospinal fluid with the automated hematology analyzer CellDyn 3500 and the urine flow cytometer UF-100.

    abstract:BACKGROUND:The counting of leukocytes and erythrocytes in cerebrospinal fluid (CSF) is still performed microscopically, e.g., using a chamber in most laboratories. This requires sufficient practical experience, is time-consuming, and may constitute a problem in emergency diagnostics. Specific automated systems for CSF ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Ziebig R,Lun A,Sinha P

    更新日期:2000-02-01 00:00:00

  • Trace metal analysis by laser-enhanced ionization in flames.

    abstract::We review atomic flame spectrometry of metals based on the selective laser-enhancement of the rate of atomic ionization. A discussion of basic principles, prior literature, and instrumentation is followed by an overview of analytical figures of merit for the technique, and a look at its present status and areas for fu...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Travis JC,Turk GC,DeVoe JR

    更新日期:1983-09-01 00:00:00

  • Concepts for the rational selection of assays to be used in monitoring therapeutic drugs.

    abstract::As the domain of monitoring therapeutic drugs rapidly expands from experimental investigations to everyday patient care, all clinical laboratory scientists, but especially clinical chemists, must learn to deal with their involvement in this expansion. First, the theoretical concepts underlying clinical pharmacology mu...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Werner M,Sutherland EW 3rd,Abramson FP

    更新日期:1975-09-01 00:00:00

  • Fluorometric assay for phospholipase A2 in serum.

    abstract::In this fluorometric assay for phospholipase A2 (EC 3.1.1.4) in serum we use the fluorescent phospholipid analog 1-octacosanyl-2-(pyren-1-yl)hexanoyl-sn-glycero-3-phosphatid yl monomethyl ester as substrate. The optimized conditions are: 28 mumol of the substrate per liter of Tris buffer (20 mmol/L, pH 7.4). The hydro...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Thuren T,Virtanen JA,Lalla M,Kinnunen PK

    更新日期:1985-05-01 00:00:00

  • Degree of unsaturation of the fatty acid chains of phospholipids influences the fluorescence polarization: implications for evaluating fetal lung maturity.

    abstract::To study the effect of fatty acid chain saturation on the fluorescence polarization assay as a measure of fetal lung maturity, we used purified phospholipids isolated from human amniotic fluid and various commercial phospholipids. We found that the fluorescence polarization value decreased as the concentration of unsa...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Dohnal JC,Bowie LJ,Burstein HJ

    更新日期:1986-03-01 00:00:00

  • Assay of carboxypeptidase N activity in serum by liquid-chromatographic determination of hippuric acid.

    abstract::We describe conditions for determining carboxypeptidase N (EC 3.4.17.3) activity by liquid chromatography. Serum (10 microL) is mixed with the artificial substrates hippuryl-L-arginine (30 mmol/L) and hippuryl-L-lysine (100 mmol/L) in 50 mmol/L 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer solution...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Hendriks D,Scharpé S,van Sande M

    更新日期:1985-12-01 00:00:00

  • Laboratory costs in the context of disease.

    abstract:BACKGROUND:To determine the contribution of laboratory costs to the overall costs of managing hospital patients with different diseases, we studied the costs of laboratory testing overall and in relation to the other costs incurred during hospitalization. METHODS:We used a database developed by the University HealthSy...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Young DS,Sachais BS,Jefferies LC

    更新日期:2000-07-01 00:00:00

  • Role of nitric oxide in cardiovascular disease: focus on the endothelium.

    abstract::Nitric oxide is a soluble gas continuously synthesized by the endothelium. This substance has a wide range of biological properties that maintain vascular homeostasis, including modulation of vascular dilator tone, regulation of local cell growth, and protection of the vessel from injurious consequences of platelets a...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Cannon RO 3rd

    更新日期:1998-08-01 00:00:00

  • Comparison of cystatin C and creatinine as predictors of cardiovascular events in a community-based elderly population.

    abstract:BACKGROUND:Reduced renal function is an established risk factor for cardiovascular events. We compared 3 measures of renal function--serum cystatin C, serum creatinine, and calculated creatinine clearance--as predictors of subsequent cardiovascular events in a community-based population of elderly individuals. METHODS...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2009.135962

    authors: Beilby J,Divitini ML,Knuiman MW,Rossi E,Hung J

    更新日期:2010-05-01 00:00:00

  • Centrifugal analyzer determination of ascorbate in serum or urine with Fe3+/Ferrozine.

    abstract::Measurement of serum ascorbate may be useful in long-term population studies because of the possible influence of ascorbate on numerous physiological factors. We describe an automated method for determining ascorbate in serum and urine by using the reduction of ferric iron by ascorbate and the formation of a color bet...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Butts WC,Mulvihill HJ

    更新日期:1975-09-01 00:00:00

  • Identification of alpha(1)-antitrypsin variants in plasma with the use of proteomic technology.

    abstract:BACKGROUND:Proteomic technology permits the investigation of genetic metabolic diseases at the level of protein expression. Changes in the expression, polypeptide structure, and posttranslational modification of individual proteins can be detected in complex mixtures of proteins. METHODS:We used high-resolution two-di...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Mills K,Mills PB,Clayton PT,Johnson AW,Whitehouse DB,Winchester BG

    更新日期:2001-11-01 00:00:00

  • Comparison of specific gravity and creatinine for normalizing urinary reproductive hormone concentrations.

    abstract:BACKGROUND:Specific gravity (SG) may perform as well as creatinine (CR) correction for adjusting urinary hormone concentrations, as well as offer some advantages. We compared the two methods and applied them to US and Bangladeshi specimens to evaluate their use in different populations. METHODS:Pearson correlations be...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2004.032292

    authors: Miller RC,Brindle E,Holman DJ,Shofer J,Klein NA,Soules MR,O'Connor KA

    更新日期:2004-05-01 00:00:00

  • Coagulation and bleeding disorders: review and update.

    abstract::Hemostasis is initiated by injury to the vascular wall, leading to the deposition of platelets adhering to components of the subendothelium. Platelet adhesion requires the presence of von Willebrand factor and platelet receptors (IIb/IIIa and Ib/IX). Additional platelets are recruited to the site of injury by release ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Triplett DA

    更新日期:2000-08-01 00:00:00

  • ISSAID/EMQN Best Practice Guidelines for the Genetic Diagnosis of Monogenic Autoinflammatory Diseases in the Next-Generation Sequencing Era.

    abstract:BACKGROUND:Monogenic autoinflammatory diseases are caused by pathogenic variants in genes that regulate innate immune responses, and are characterized by sterile systemic inflammatory episodes. Since symptoms can overlap within this rapidly expanding disease category, accurate genetic diagnosis is of the utmost importa...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1093/clinchem/hvaa024

    authors: Shinar Y,Ceccherini I,Rowczenio D,Aksentijevich I,Arostegui J,Ben-Chétrit E,Boursier G,Gattorno M,Hayrapetyan H,Ida H,Kanazawa N,Lachmann HJ,Mensa-Vilaro A,Nishikomori R,Oberkanins C,Obici L,Ohara O,Ozen S,Sarkisian T

    更新日期:2020-04-01 00:00:00

  • Immunoluminometric assay for measurement of the C-terminal endothelin-1 precursor fragment in human plasma.

    abstract:BACKGROUND:Endothelin-1 (ET-1), a potent vasoconstrictor, is difficult to measure because of its instability and its binding to receptors and plasma proteins. We report a rapid, robust way to indirectly quantify ET-1 release by measuring the C-terminal ET-1 precursor fragment (CT-proET-1) without an extraction step. M...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2005.065581

    authors: Papassotiriou J,Morgenthaler NG,Struck J,Alonso C,Bergmann A

    更新日期:2006-06-01 00:00:00

  • Highly sensitive enzyme immunoassay of proinsulin immunoreactivity with use of two monoclonal antibodies.

    abstract::A highly sensitive two-site sandwich ELISA measuring total proinsulin immunoreactive material in serum or plasma was developed. The assay was based on two monoclonal antibodies, an anti-C-peptide antibody bound to a microtest plate and a biotin-labeled anti-insulin antibody. The detection limit (3 SD above zero value)...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Kjems LL,Røder ME,Dinesen B,Hartling SG,Jørgensen PN,Binder C

    更新日期:1993-10-01 00:00:00

  • Enzyme-linked immunosorbent assay for 17 alpha-hydroxyprogesterone in dried blood spotted on filter paper.

    abstract::In this rapid, cost-effective, enzyme-linked immunosorbent assay for 17 alpha-hydroxyprogesterone (17-OHP) eluted from dried blood spotted on filter paper, second antibody is coated onto the microwell plate and horseradish peroxidase (EC 1.11.1.7) is the label enzyme. Antiserum to 17-OHP was prepared by using 4-(2-car...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Maeda M,Arakawa H,Tsuji A,Yamagami Y,Isozaki A,Takahashi T,Haruki E

    更新日期:1987-06-01 00:00:00

  • Thyroxine-binding globulin capacity and concentration evaluated from blood spots on filter-paper in a screening program for neonatal hypothyroidism.

    abstract::We describe a simple method for evaluating thyroxine-binding globulin capacity and concentration from a single 1-cm blood spot on filter-paper used in a screening program for neonatal hypothyroidism. This method permits prompt diagnosis of about 90% of the infants with thyroxine-binding globulin deficiency in our abno...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Dussault JH,Morissette J,Letarte J,Guyda H,Laberge C

    更新日期:1980-03-01 00:00:00

  • Estrogen and progesterone receptors in breast cancer microsamples simultaneously quantified by enzyme-ligand immunoassay.

    abstract::A new method (enzyme-ligand immunoassay, ELIA) is described for the estimation of estrogen (ER) and progesterone (PR) receptors in microsamples of human breast cancer tissue. The technique, based on the nonisotopic measurement of receptor-bound estradiol and progesterone, involves three steps: (a) simultaneous saturat...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Krambovitis E,Hatzidakis G,Hatzoglou A,Romain S,Durand A,Stefanakis A,Castanas E

    更新日期:1995-01-01 00:00:00

  • Improved liquid chromatographic/immunoassay of digoxin in serum.

    abstract::This HPLC/immunoassay procedure measures digoxin in serum with no interference from digoxin metabolites or digoxin-like factors. We used solid-phase (C18 and Diol) extraction, a C18 column with a tetrahydrofuran/water mobile phase, and final quantification by fluorescence polarization immunoassay. Deslanoside and gito...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Stone JA,Soldin SJ

    更新日期:1988-12-01 00:00:00

  • Automated analysis for plasma epinephrine and norepinephrine by liquid chromatography, including a sample cleanup procedure.

    abstract::We describe a fully automated method of analysis for plasma epinephrine and norepinephrine by use of a two-column system of "high-performance" liquid chromatography. Catecholamines in deproteinized plasma are purified on the first (preparation) column, then transferred automatically to the second (analytical) column i...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Yamatodani A,Wada H

    更新日期:1981-12-01 00:00:00

  • Proposed reference method for iron in serum used to evaluate two automated iron methods.

    abstract::The manual Reference Method of the Centers for Disease Control for serum iron (CDC/RM/Fe) and a semiautomated adaptation of it were used to evaluate two working methods: one, a detergent solubilization procedure for the Roche Cobas-Bio analyzer, the other, the Kodak Ektachem 700 procedure, based on dry-film technology...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Valcour AA,Krzymowski G,Onoroski M,Bowers GN Jr,McComb RB

    更新日期:1990-10-01 00:00:00

  • Measurement of thyroglobulin by liquid chromatography-tandem mass spectrometry in serum and plasma in the presence of antithyroglobulin autoantibodies.

    abstract:BACKGROUND:Measurement of serum thyroglobulin (Tg) is used to monitor patients after treatment for differentiated thyroid carcinoma (TC). Difficulty in using Tg as a biomarker of the recurrence of TC in many patients stems from the presence of endogenous anti-Tg autoantibodies (Tg-AAbs), which can interfere with immuno...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2012.195594

    authors: Kushnir MM,Rockwood AL,Roberts WL,Abraham D,Hoofnagle AN,Meikle AW

    更新日期:2013-06-01 00:00:00

  • Production of murine hybrid-hybridomas secreting bispecific monoclonal antibodies for use in urease-based immunoassays.

    abstract::To produce bispecific antibodies (BiAbs) for enzyme immunoassay (EIA) to replace antibody-enzyme conjugates, we developed a panel of 8-azaguanine/ouabain-resistant anti-urease variant hybridoma cell lines for use in hybridoma-hybridoma fusions. These variants represent mouse immunoglobulin subclasses IgM, IgG1, IgG2a,...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Takahashi M,Fuller SA

    更新日期:1988-09-01 00:00:00

  • Enzymic inhibition assay for methotrexate with a discrete analyzer, the ABA-100.

    abstract::We adapted an inhibition assay for methotrexate, involving dihydrofolate reductase from bovine liver, for use with a discrete analyzer (the ABA-100). The analyzer was used both for dilution and a 5-min pre-incubation of the sample with NADPH--enzyme reagent, and for the assay itself. The standard curve was linear betw...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Brown LF,Johnson GF,Witte DL,Feld RD

    更新日期:1980-02-01 00:00:00

  • Seven different assays of hyaluronan compared for clinical utility.

    abstract::To compare six assays of hyaluronan (hyaluronic acid; HYA) in serum, developed in different laboratories, we analyzed 10 samples from each of three groups: healthy persons, patients with primary biliary cirrhosis, and patients with rheumatoid arthritis. All the assays are based on the use of affinity proteins specific...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Lindqvist U,Chichibu K,Delpech B,Goldberg RL,Knudson W,Poole AR,Laurent TC

    更新日期:1992-01-01 00:00:00

  • Relation of lipoprotein(a) in 11- to 19-year-old adolescents to parental cardiovascular heart disease.

    abstract::We studied several risk factors in relation to parental cardiovascular heart disease: total cholesterol, triglycerides, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, apolipoprotein (apo) A-I, apo B, and lipoprotein(a) [Lp(a)] were determined in 322 serum samples (43 from subjec...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Vella JC,Jover E

    更新日期:1993-03-01 00:00:00