bluuebunny/crossref_metadata_2025 huggingface dataset:
Otologic side effects of drugs
Authors: Lee C A , Mistry D , Uppal S , Coatesworth A P
Published: May 2005

Otolaryngologic symptoms are common and patients presenting to the otolaryngologist will often be taking drugs for the treatment of unrelated diseases. As a doctor, one must not forget the potential of these drugs to cause otologic side effects and, in some cases, to be the cause of the presenting symptom.

We performed a comprehensive search of the British National Formulary and Electronic Medical Compendium websites to classify the otologic side effects caused by drugs. Not all the data were found at both websites. We have compiled all the data together, subclassified them and produced a review of the otologic side effects of drugs, in table form.
Activation of PAR4 Induces a Distinct Actin Fiber Formation via p38 MAPK in Human Lung Endothelial Cells
Authors: Fujiwara Masakazu , Jin Enjing , Ghazizadeh Mohammad , Kawanami Oichi
Published: May 2005

Protease-activated receptors (PARs) are multifunctional G protein–coupled receptors. Among the four existing PARs, PAR4 is preferentially expressed in the human lung tissue. However, the function of PAR4 has not been defined in the lung endothelial cells. Because PAR1-mediated cellular effects are deeply related to the morphological changes, we focused on the actin fiber and p38 mitogen-activated protein kinase (MAPK) signaling involved in actin polymerization to elucidate the role of PAR4. RT-PCR and Western blot analyses identified PAR4 expression in human pulmonary artery endothelial cells and in human microvascular endothelial cells from lung. We then examined the changes in actin fibers in endothelial cells treated with PAR4-activating peptide. PAR1-activating peptide was used for comparison. Activation of PAR4 and PAR1 by their corresponding peptides induced actin fiber formation; however, the actin filaments were broadly bundled in PAR4 as compared with the ringlike actin filaments in PAR1 activation. Correspondingly, the magnitude of p38 MAPK phosphorylation was different between cells treated with PAR4 and PAR1, with PAR4-activating peptide showing a significantly higher sensitivity to p38 MAPK inhibitor, SB203580. Taken together, these results demonstrate that activation of PAR4 results in the formation of actin fiber distinct from that by PAR1 activation, suggesting PAR4 may play specific roles in the lung endothelial cells.
Establishment of Stable GH3 Cell Line Expressing Enhanced Yellow Fluorescein Protein-Growth Hormone Fusion Protein
Authors: Matsuno Akira , Mizutani Akiko , Itoh Johbu , Takekoshi Susumu , Nagashima Tadashi , Okinaga Hiroko , Takano Koji , Osamura R. Yoshiyuki
Published: May 2005

To investigate, in real time, the transport and secretion of pituitary hormone, we have developed an experimental pituitary cell line, GH3 cell, which has secretory granules of growth hormone (GH) linked to enhanced yellow fluorescein protein (EYFP). This stable GH3 cell secretes secretory granules of GH linked to EYFP on stimulation by Ca2+ influx or Ca2 release from storage. This GH3 cell will be useful for the real-time visualization of the intracellular transport and secretion of GH.
Mucin Granules are in Close Contact with Tubular Elements of the Endoplasmic Reticulum
Authors: Perez-Vilar Juan , Pedrosa Ribeiro Carla M. , Salmon Wendy C. , Mabolo Raean , Boucher Richard C.
Published: May 2005

Live cell imaging methods were used to characterize goblet cells expressing a MUC5AC domain fused to enhanced green fluorescent protein that labels the granule lumen. Golgi complex and endosome/lysosome elements largely resided in the periphery of the granular mass. On the contrary, a tubular meshwork of endoplasmic reticulum (ER) was in close contact with the mucin granules. This meshwork could be identified in fixed native human bronchial goblet cells labeled with an anti-calreticulin antibody. The potential biological significance of this ER network for mucin secretion is discussed.
Ultrastructural Studies of Human Basophils and Mast Cells
Authors: Dvorak Ann M.
Published: May 2005

Ultrastructural studies of human mast cells (HMCs) and basophils (HBs) are reviewed. Sources of HMCs include biopsies of tissue sites and in situ study of excised diseased organs; isolated, partially purified samples from excised organs; and growth-factor-stimulated mast cells that develop de novo in cultures of cord blood cells. Sources of HBs for study include partially purified peripheral blood basophils, basophils in tissue biopsies, and specific growth factor-stimulated basophils arising de novo from cord blood cells. The ultrastructural studies reviewed deal with identity, secretion, vesicles, recovery, and synthesis issues related to the biology of these similar cells.
Thermisches Verhalten und Struktur von Copolymeren der Maleinsäure‐bzw. Fumarsäurehalbester
Authors: Rätzsch M. , Steinert V. , Seiler S. , Büttner B.
Published: May 2005

Das thermische Verhalten von Maleinsäurehalbester‐Copolymeren, die durch Veresterung von Maleinsäureanhydrid‐Copolymeren hergestellt wurden, und der Copolymere der Malein‐ sowie Fumarsäurehalbester wird untersucht. Daraus werden Rückschlüsse auf die Struktur der Copolymere gezogen. Die Schwierigkeiten bei der experimentellen Durchführung der Cyclisierungsreaktion in Lösung, die zur Umsetzung in der Festphase geführt haben, werden beschrieben.
Zur Lichtstreuung an Polymerdispersionen. Teil 5. Theoretische Untersuchungen zur Auswertung von Streulichtmessungen im Bereich der Mie‐Streuung
Authors: Hartmann M. , Dautzenberg H.
Published: May 2005

Ein neues Verfahren zur Auswertung von experimentellen Streukurven im Bereich der Mie‐Streuung auf Basis der Mie‐Theorie unter Einbeziehung von Näherungsformeln der geometrischen Optik wird vorgestellt. Mit Hilfe dieser Methode ist es möglich, aus einem experimentell ermittelten Wertepaar von Streuintensität beim Streuwinkel 0° und Anfangsanstieg der Streukurve die dem gegebenen streuenden System entsprechenden Verteilungsparameter zu finden. Dies ist auch dann der Fall, wenn mehrere Zuordnungen (Mehrdeutigkeit der Mie‐Streuung) existieren.
Copolymerization of acrylonitrile with some transition metal acrylates. II. Determination of monomer reactivity ratios in radical copolymerization of acrylonitrile with cobalt, nickel and copper acrylates in dimethylformamide
Authors: Czerniawski T. , Wojtczak Z.
Published: May 2005

The results of the copolymerization of acrylonitrile with cobalt, nickel and copper acrylates in dimethylformamide solution at 60°C with AIBN as an initiator are presented. The copolymerization parameters are discussed.
Synthese und einige physiko‐chemische Eigenschaften makrovernetzter isoporöser Styrenpolymere mit Vernetzungsbrücken vom Diphenylmethantyp
Authors: Cjurupa M. P. , Lalaev V. V. , Davankov V. A.
Published: May 2005

Die makrovernetzten isoporösen Polymere wurden durch Vernetzung von Polystyren in Lösung mit Dimethylformal in Gegenwart von Friedel‐Crafts‐Katalysatoren erhalten. Es wurden die Reaktionsbedingungen ermittelt (20°C, 5 h, 1 mol SnCl4 auf 1 mol Methylal), bei denen das Vernetzungsmittel praktisch vollständig zur Bildung von Diphenylmethanbrücken verbraucht wird. Die erhaltenen Polymere können sowohl in Lösungs‐ als auch Fällmitteln für lineares Polystyren quellen. Die innere spezifische Oberfläche stark vernetzter Proben erreicht 900 m2/g.
Über das mechanische schmelzen des supergitters von blockcopolymeren
Authors: Skazka V. S. , Ručin A. E. , Frenkel S. Ja. , Vilesov A. D.
Published: May 2005

Die Temperaturabhängigkeit des photoelastischen Koeffizienten ϵ von ternären Blockcopolymeren aus Styren und Butadien mit verschiedenen Komponentenverhältnissen wird untersucht. Hierbei wird ein starker Einfluß des optischen Formeffektes auf die Abhängigkeit ϵ(T) nachgewiesen. Es wird gezeigt, daß die Störung der Regularität des Superkristallgitters des Blockcopolymers bei Temperaturen erfolgt, die erheblich oberhalb der höchsten Glastemperatur der Komponenten liegen. Dieser Prozeß wird als mechanisches „Schmelzen”︁ des Supergitters betrachtet.
Fraktographie von Polyamidfäden. I. Einfluß von Verstrecktemperatur, Verstreckgeschwindigkeit, Umgebungsmedium und Vorbehandlung
Authors: Hinrichsen G. , Altemeier G. , Renger M.
Published: May 2005

Ein unpigmentiertes Polyamid‐Filamentgarn wird unter Variation von Verstrecktemperatur, ‐geschwindigkeit und ‐medium bis zum Bruch verdehnt. Die entstandenen Bruchflächen werden mittels Rasterelektronenmikroskopie auf ihre Morphologie hin untersucht. Es werden für die jeweiligen Verstreckbedingungen charakteristische Bruchmorphologien nachgewiesen. Die Brüche starten durch ein Aufplatzen des Fasermantels.
InGaAlN and II–VI Systems for Blue–Green Light‐Emitting Devices
Authors: Matsuoka Takashi
Published: May 2005

Recent progress in research on InGaAlN and II–VI systems has been remarkable, and this paper reviews the properties of these materials from the viewpoint of making reliable light‐emitting devices. The growth of these materials by molecular beam epitaxy (MBE) and metalorganic vapor phase epitaxy (MOVPE) is also reviewed, and the current status of both materials is reviewed with respect to light‐emitting devices.
Visualization of immobilization in shear bands by NMR imaging
Authors: Weigand Frank , Wiesner Ulrich , Spiess Hans Wolfgang
Published: May 2005

A new 1H‐NMR imaging method, in which the relaxation time provides the contrast, is demonstrated to be suitable for investigating the effect of shear bands—strongly localized inhomogeneities of the plastic deformation—on the local mobility of polymeric materials in the solid state. The immobilization of a shear band in cold‐drawn polycarbonate (PC) is visualized for the first time. In addition to a description of the experiment, results for PC are reported, showing, for example, that the phenylene motion is more affected by the strain than the methyl motion.
Oligomeric arsenic nitrides
Authors: Schmid G. , Hättmann Katja
Published: May 2005

Oligomeric AsN particleshave been synthesized as a first step towards forming polymeric (AsN)x, which is expected to be the arsenic nitride with the most important applications. The synthesis is described and the characterization of the resulting oligomers by X‐ray powder diffraction, mass spectrometry and DSC is presented which leads to the proposed structure of (AsN)12(AsCl2)6Cl6shown in the Figure.

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Persistent photoinduced electron transfer from functionalized dendrimers to Buckminsterfullerene
Authors: Janssen René A. J. , Jansen Johan F. G. A. , van Haare John A. E. H. , Meijer E. W.
Published: May 2005

Functionalized poly(propylene imine) dendrimers are efficient electron donors in photoinduced electron transfer reactions to Buckminsterfullerene in solution. The photoinduced electron transfer reaction produces persistent fullerene radical anions at room temperature even with light switched off, as evidenced from ESR spectroscopy. The reaction exhibits a remarkable size‐effect: persistent electron transfer is observed only for dendrimer generations G ≥ 2.
A new approach to polybenzoid networks
Authors: Mindach Lutz , Müllen Klaus
Published: May 2005

High‐carbon materials, which contain a high density of strong, directed CC bonds, are expected to exhibit some of the outstanding properties of diamond and graphite, e.g., hardness, thermal and electrical conductivity. The thermolysis of biphenylene‐containing monomers is demonstrated to offer a promising new approach to polybenzoid networks such as that shown in the Figure.

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Peripheral nerve abnormality in multiple sclerosis
Authors: Pollock Martin , Calder Christopher , Allpress Stephen
Published: May 2005

Biopsy specimens of sural nerve from 10 patients with multiple sclerosis (MS), of whom only 1 was severely disabled, were assessed by morphometric techniques and compared to nerves from age‐matched controls. The frequency of abnormal teased fibers was increased in MS nerve, with many intenodes showing at least a 50% reduction in myelin thickness. Myelinated nerve fiber densities were not significantly different in MS and control nerves. Regression analysis of axonal area on number of myelin lamellae indicated a generalized reduction of myelin lamellae in this disorder. It is suggested that peripheral myelin may be involved in multiple sclerosis.

Supported by the New Zealand Neurological Foundation, the Medical Research Council, and the National, Otago, and Canterbury Multiple Sclerosis Societies.
“Inverse latent” macro square‐wave jerks and macro saccadic oscillations
Authors: Dell'Osso L. F. , Abel L. A. , Daroff R. B.
Published: May 2005

A patient recovering from an acute encephalopathy demonstrated several ocular motor disturbances reflecting cerebellar and brainstem dysfunction. Two of these, macro square‐wave jerks and macro saccadic oscillations, have never been reported in the same patient. The macro square‐wave jerks disappeared with monocular viewing, a previously undescribed “inverse latent” characteristic.
Diencephalic syndrome and von recklinghausen's disease
Authors: Adornato Bruce , Berg Bruce
Published: May 2005

Two infants with von Recklinghausen's disease developed signs and symptoms of the diencephalic syndrome. Pathological examination confirmed the presence of hypothalamic tumors in both. The concurrence of these two entities, although previously unreported, is not unexpected given the incidence of intracranial tumor in von Recklinghausen's disease and the dinecephalic syndrome.
The effect of two bisphosphonates on human neutrophil chemiluminescence and myeloperoxidase activity
Authors: Kowolik M. J. , Hyvönen P. M. , Sutherland R. , Raeburn J. A.
Published: May 2005

In order to assess the importance of chlorine in a drug molecule as an influence on myeloperoxidase‐mediated inflammatory cell functions, the effect of the chlorinated bisphosphonate, clodronate, on human neutrophil chemiluminescence and myeloperoxidase (MPO) activity was compared to the non‐chlorinated structural analogue, etidronate. The results suggested that the presence of chlorine may be important to the enhancement of MPO activity. In addition both drugs manifested low toxicity and both of these observations may have relevance to host defence.
Chemiluminescent assay of enzymes using proenhancers and pro‐anti‐enhancers
Authors: Kricka Larry J. , Schmerfeld‐Pruss Deborah , Edwards Brooks
Published: May 2005

Enhanced chemiluminescent assays for hydrolase enzymes have been developed using proehancer and pro‐anti‐enhancer substrates. Alkaline phosphatase is measured using disodium para‐iodophenyl phosphate (proenhancer) which is converted to para‐iodophenol and this in turn enhances the light emission from the horseradish peroxidase catalysed chemiluminescent oxidation of luminol by peroxide. An alternative strategy uses para‐nitrophenyl phosphate which is converted by alkaline phosphatase to para‐nitrophenol which inhibits the enhanced chemiluminescent reaction. The detection limit for the enzyme using the proenhancer and pro‐anti‐enhancer assays was 100 attomoles and 1 picornole, respectively. The proenhancer strategy was effective in assays for beta‐D‐galactosidase, beta‐D‐glucosidase and aryl sulfatase. A limited comparison of the proenhancer and a conventional colorimetric assay for an alkaline phosphatase label in an enzyme immunoassay for alpha‐fetoprotein showed good agreement.
Attenuation of 4‐I‐phenol‐enhanced chemiluminescence by non‐enhancer phenols
Authors: Candy Thomas E. G. , Jones Peter
Published: May 2005

The intensity of 4‐I‐phenol‐enhanced chemiluminescence from the luminol‐H2O2‐horseradish peroxidase system is markedly attenuated in the presence of low concentrations of non‐enhancer phenols. Under the conditions studied, the effect is not associated with competition between 4‐I‐phenol and non‐enhancer phenol for the enzyme intermediates, Compounds I and II, but involves a competition between non‐enhancer phenol and luminol most probably for the 4‐I‐phenoxy radical.
Fluorescence high performance liquid chromatographic determination of 3α‐hydroxysteroids in urine of 21‐hydroxylase deficiency
Authors: Kawasaki Takao , Maeda Masako , Tsuji Akio
Published: May 2005

A fluorescence high performance liquid chromatographic method using an immobilized 3α‐hydroxysteroid dehydrogenase column as a post‐column enzymatic reactor was developed for the determination of corticosteroid metabolites in the urine of subjects with congenital adrenal hyperplasia. 3α‐Hydroxysteroids, such as pregnanetriol, pregnanediol and pregnanetriolone, in the eluate from μ‐Bondapak phenyl column (300 × 3.9 mm I.D.) using 0.05% ammonium phosphate buffer (pH 7.1)–acetonitrile–methanol (100: 55: 15) as the mobile phase was mixed with NAD+ solution in the enzyme column at 30 °C to generate NADH, which was monitored by a fluorophotometric detector. Each steroid was measured at the 2.5 μg/dl at the highest sensitivity of the detector. The mean recoveries and reproducibilities were 91.5–108.2% with 0.9–6.5% (CV%).
Early identification of isovaleric aciduria using photodiode array detection for liquid chromatographic profiling of urinary carboxylic acids
Authors: Buchanan D. N. , Thoene J. G.
Published: May 2005

The 190 nm high performance liquid chromatographic photodiode array profiling of the urinary carboxylic acids of the first urine of a newborn affected with isovaleric aciduria afforded an abnormal peak at 27.8 min. This peak was greatly increased in the carboxylic acid profiling of the 14 h urine sample from the same infant. Isolation of this peak by fraction collecting; solvent extraction of the eluent; trimethylsilyl derivatization of the residue and gas chromatographic/mass spectrometric analysis identified the compound as isovalerylglycine. Correlation of the 190 nm absorbance of isovalerylglycine (y) with concentration (x) afforded a least squares curve: y = 476.4x – 13.72 (r = 0.99) run‐to‐run variation 6.92%; day‐to‐day variation 8.88%; with a minimum detectable concentration of 25 μg/ml.
Combined use of lectin affinity chromatography and endo‐β‐galactosidase to study polylactosamine sequences isolated from haemopoietic cell surfaces
Authors: Morris Andrew J. , Gallagher John T. , Dexter T. Michael
Published: May 2005

The trypsin‐sensitive glycopeptides from cell surfaces of a multipotential murine haemopoietic cell line (DE) have been studied using serial lectin affinity chromatography on columns of immobilized lentil lectin (LCA), concanavalin A (Con A), and wheat‐germ agglutinin (WGA). WGA‐binding material consisted of glycopeptides that failed to bind to LCA and Con A. Step elution from the WGA‐column with 0.01‐, 0.1‐, 0.5‐ and 1.0 M N‐acetyl‐D‐glucosamine yielded four affinity classes of glycopeptide (WGA‐W, WGA‐I, WGA‐S and WGA‐SS respectively). WGA‐W, WGA‐I and WGA‐S contained both alkali‐stable (N‐linked) and alkali‐labile (O‐linked) carbohydrate on high molecular weight glycopeptides. The WGA‐SS fraction contained only N‐linked carbohydrate. N‐linked glycopeptides isolated from each WGA‐binding class differed in molecular size, relative N‐acetylneuraminic acid content and affinity for Ricinus communis 120 agglutinin. endo‐β‐Galactosidase digestion showed that these glycopeptides contained polylactosamine‐type glycans. Gel filtration profiles of the enzyme treated materials were different for each WGA‐binding population suggesting variation in branching patterns and/or substitution with fucose residues. Affinity chromatography has shown that the WGA binding molecules are the major glycopeptide group at DE cell surfaces.
A simple quantitative HPLC assay for ifosfamide in biological fluids
Authors: Margison Jennifer M. , Wilkinson P. M. , Cerny T. , Thatcher N.
Published: May 2005

A high performance liquid chromatography method is described for measuring Ifosfamide (I) in human serum. This involves solvent extraction, reverse phase HPLC and UV detection at 190 nm. Standard curves of peak height × detector sensitivity versus I concentration in serum were linear with a lower limit of detection of 100 ng/ml. Authentic 14C‐labelled I cochromatographed with standard I and with I found in serum from treated patients. The concentration‐time curves of I determined by both HPLC and gas chromatography were indistinguishable. We conclude that this method is suitable for determining I pharmacokinetics in biological specimens.
Biosynthesis of bloodgroup I and i antigens. A sensitive and specific assay of UDP‐GlcNAc:β‐galactoside β1 → 3‐N‐acetylglucosaminyltransferase activity in hematopoietic cells by HPLC
Authors: Koenderman Anky H. L. , Loppen Piet L. , Marinus Lian A. M. , van den Eijnden Dirk H.
Published: May 2005

A sensitive HPLC method for the assay of UDP‐GlcNAc:β‐galactoside β1 → 3‐N‐acetylglucosaminyltransferase activity was developed. Using lactose as an acceptor, the formation of the product GlcNAcβ1 → 3Galβ1 → 4Glc can be determined without interference by substrates resulting from enzymatic and chemical breakdown of the donor substrate UDP‐GlcNAc. The method is very specific since products of other transferase reactions, which potentially may be formed in the incubations in vitro, elute at positions different from that of GlcNAcβ1 → 3Galβ1 → 4Glc.

By use of this assay method it could be demonstrated that normal and malignant hematopoietic cells and cell‐lines, with the exception of erythrocytes and reticulocytes, contain β1 → 3‐N‐acetylglucosaminyltransferase activity.
Measurement of penbutolol and 4‐hydroxypenbutolol in plasma or serum by HPLC
Authors: Bhamra R. K. , Flanagan R. J. , Holt D. W.
Published: May 2005

A simple HPLC method for penbutolol and 4‐hydroxypenbutolol assay has been developed. Plasma or serum (200 μl) is vortex‐mixed (30 s) with Tris solution (2 M, pH 10.6) containing an internal standard (50 μl) and methyl t‐butyl ether (200 μl). After centrifugation, the extract (100 μl) is analysed using an unmodified silica column (250 × 5 mm ID) and iso‐octane–methanol–methyl t‐butyl ether (55:25:20) containing ammonium perchlorate (10 mM, pH 5.7) as eluent and with fluorescence detection. No interference has been encountered and the limit of accurate measurement for both compounds is 5 μg/l.
A simple method for the purification of dipeptidyl aminopeptidase IV from porcine kidney using HPLC
Authors: Hutchinson D. W. , Woolfitt A. R.
Published: May 2005

Depeptidyl aminopeptidase IV (E.C. 3.4.14.5) has been purified to homogeneity using “soft‐column” chromatography on Sephadex G‐200, DEAE‐cellulose, gly‐pro‐AH‐Sepharose and finally HPLC on TSK G‐3000SW. The purification by HPLC is fast and gives a better yield of pure enzyme than procedures which have been described previously. The enzyme obtained in this manner has a high specific activity (86 U/mg using a saturating level of gly‐pro‐4‐nitroanilide as substrate) and was free of contaminating peptidase activities. Our preparation of DAP IV is suitable for peptide sequencing studies by fast atom bombardment mass spectrometry.
Solid phase extraction and isocratic separation of urinary porphyrins by HPLC
Authors: Kotal Petr , Jirsa Milan , Martásek Pavel , Kordač Václav
Published: May 2005

A method for determining urine porphyrins by HPLC is described. In the preliminary step, porphyrins are purified in high yields and concentrated by low pressure reverse‐phase chromatography on C18 (octadecylsilane bonded silica) cartridge. Porphyrins are stable for 10 days after adsorption on C18 cartridge. The separation of porphyrin esters is performed on an aminopropyl‐bonded silica column with an eluting system containing n‐heptane and ethyl acetate. The system enables rapid isocratic separation of porphyrin methyl esters with high selectivity. The simplicity and reproducibility of the whole procedure allows its application to the routine analysis of urinary porphyrins in the clinical laboratory.
Red cell zinc protoporphyrin and protoporphyrin by HPLC with fluorescence detection
Authors: Rossi E. , Garcia‐Webb P.
Published: May 2005

A high performance liquid chromatographic (HPLC) method was developed for the determination of zinc protoporphyrin (ZnPP) and protoporphyrin (PP) in whole blood. After adding the blood to dilute acetic acid, ZnPP and PP were extracted with dimethyl sulfoxide–acetone containing mesoporphyrin as internal standard. Following evaporation of the acetone, the haemin‐free extract was analysed by HPLC. ZnPP and PP were separated on a reversed‐phase column and quantitated by measuring fluorescence peak areas. The extraction method is simple, and applicable to batch analysis, and the HPLC separation is rapid and repoducible. The coefficient of variation for ZnPP was 5.6% and 3.3% for total red cell porphyrin levels of 3.5 and 10.2 μmol per litre RBC respectively. Results are discussed in patients with erythrohepatic protoporphyria, lead exposure, iron‐deficiency and non‐specifically elevated total red cell porphyrins.
HPLC determination of orthophosphate coexisting with large amounts of organophosphates in biological samples
Authors: Koshiishi Ichiro , Imanari Toshio
Published: May 2005

A simple and accurate HPLC method for the determination of Orthophosphate in the presence of large amounts of organophosphates is described. The method is based on the formation and separation of the molybdenum Orthophosphate complex. In order to prevent the hydrolysis of organophosphates, the sample was deproteinized with silicotungstate in acetate buffer (pH 4.0) under ice‐cooling and then treated with ammonium molybdate in maleate buffer (pH 7.0). The sample was injected onto Styragel 60 Å column (5 mm ID × 100 mm) with 38% (v/v) acetonitrile containing 0.3 M sulfuric acid as eluent. Detection was at 310 nm. The method was applied to the determination of Orthophosphate in liver, kidney, spleen and mouse blood.
Determination of free and total catecholamines in human urine by HPLC with fluorescence detection
Authors: Nohta Hitoshi , Mitsui Akane , Umegae Yoshihiko , Ohkura Yosuke
Published: May 2005

A simple and highly sensitive method for the determination of free and total (free + conjugated) catecholamines (norepinephrine, epinephrine and dopamine) in human urine is described which employs HPLC with fluorescence detection. Conjugated catecholamines (sulfate form) are hydrolyzed by a sulfatase‐mediated reaction to the corresponding free amines. After cation exchange chromatography on a Toyopak IC‐SPS cartridge, catecholamines and isoproterenol (internal standard) in urine samples were converted into the corresponding fluorescent compounds by reaction with 1, 2‐diphenylethylenediamine. These compounds were separated within 8 min on a reversed phase column with isocratic elution using a mixture of water, methanol and acetonitrile containing a Tris‐hydrochloric acid buffer (pH 7.0). The detection limit for each catecholamine is ca 2 fmol per 100 μL injection volume.
Analysis of p‐aminobenzoic acid and its metabolites in urine and plasma by ion pair HPLC in the NBT‐PABA pancreatic function test
Authors: Hung C. T. , Perrier D. G. , Schicker A. M. , Zoest A. R.
Published: May 2005

An ion pair HPLC method which can simultaneously detect the major metabolites of an exocrine pancreatic function testing agent (NBT‐PABA) in plasma and urine has been developed. This assay has been applied to a pharmacokinetic study of PABA and its metabolites in 3 healthy adult volunteers following the oral administration of 1 g NBT‐PABA. The 6 h testing period currently used to collect urine following the NBT‐PABA ingestion is adequate for the recovery of PABA and its metabolites. Plasma determination of these compounds may provide an improved evaluation of the pancreatic performance in patients with abnormal liver or kidney function.
Simultaneous determination of gastrodin and its metabolite by HPLC
Authors: Liu Ke‐Xin , Han Guo‐Zhu , Chang Ya‐Lun , Su Cheng‐Ye , Tang Nan‐Yuan , Chen Yu‐Ren
Published: May 2005

A rapid and specific HPLC method for the simultaneous determination of gastrodin and its metabolite gastrodigenin (p‐hydroxybenzyl alcohol) in rat plasma, bile, liver, urine and faeces is described. The separation was achieved by using a reversed phase column (YWG‐C18) eluted with methanol‐water (2.5:97.5 v/v). Phloroglucinolum was used as internal standard and the peaks were detected at UV 221 nm. The protein precipitation with ethanol was a very simple and rapid method for sample preparation. The gastrodin and gastrodigenin were quantitated by measuring the peak‐height ratios. There was a linear concentration range of 10–320 μg/mL in the assay for both compounds. The coefficients of variation (within‐day) for samples spiked with gastrodin and gastrodigenin were 2.94% and 3.08%, respectively. The method demonstrated a high specificity and was suitable for use in pharmacokinetic studies.
Rapid purification of arrowhead proteinase inhibitors by high performance hydrophobic interaction chromatography on a PEG bonded phase column
Authors: Chang Jen‐Ping , Yuan Yun
Published: May 2005

A new hydrophobic interaction HPLC column is used for the rapid purification of proteinase inhibitors isolated from arrowhead. The inhibitors, partially purified by DEAE‐cellulose column chromatography, are resolved into three components with a mobile phase gradient of decreasing salt concentration from 1.1 M ammonium sulfate in 0.01 M phosphate buffer to phosphate buffer alone. This new HPLC column is found to be very useful for rapid, semipreparative purification of hydrophobic protein and sample loading of up to 1.6 mg of inhibitors can be fully resolved on an analytical column.
HPLC measurement of penticainide and desalkylpenticainide in biological fluids
Authors: Walker S. , Flanagan R. J. , Holt D. W.
Published: May 2005

A simple HPLC method has been developed to measure the antiarrhythmic agent penticainide and its N‐desalkyl metabolite in biological fluids. Solvent extraction of a small (200 μL) sample volume with direct analysis of the extract is used to measure the plasma and urinary concentrations of these compounds attained during chronic therapy, although a larger sample volume (1.0 mL) and prior concentration of the extract are required for single oral dose work. In each case chromatographic analysis is performed using a microparticulate (5 μm) silica column and methanolic ammonium perchlorate (10 mM, pH 6.7) as eluent with UV detection (260 nm). No endogenous sources of interference have been encountered and potential interference from other drugs is minimal.
Determination of adriamycin and its fluorescent metabolites in human plasma by HPLC
Authors: Mahdadi R. , Lhermitte M. , Lafitte J. J.
Published: May 2005

We describe a method for measuring adriamycin and its metabolites, adriamycinol and adriamycinone in plasma, using reversed phase HPLC and fluorescence detection. The lower limit of detection is approximately 1 ng/mL for each compound. An extraction technique for serum is described which is capable of an almost equal recovery (>93%) of adriamycin and metabolites without interference from endogenous components of plasma and from other common drugs. Within‐day and day to day coefficients of variation are estimated.
The use of bond‐elut for the estimation of serum bile pigments bonded covalently to albumin
Authors: McKavanagh S. M. , Billing B. H.
Published: May 2005

A simple and precise method has been devised for the quantitation of biliprotein (δ‐bilirubin or albumin bound bilirubin) in serum. In the presence of caffeine/benzoate, Bond‐Elut (C8, 200 mg) extracts unconjugated and conjugated bilirubin but not pigments that are covalently bonded to albumin which pass through the column and can be quantitated by a standard diazo method. Following elution from the Bond‐Elut column with methanol‐acetonitrile (50: 50, v/v) unconjugated and conjugated bilirubin can be quantitated either as total pigments or individually by HPLC.
Identification of isoprenoid‐type components in human expired air: A possible shunt pathway in sterol metabolism
Authors: O'Neill Hugh J. , Gordon Sydney M. , Krotoszynski Boguslaw , Kavin Hymie , Szidon Jan P.
Published: May 2005

Expired air samples have been analyzed from three groups of human subjects (normal, liver dysfunction, lung cancer) and the baboon (Papio anubus). Of the several hundred compounds present, three compounds were of particular interest due to their structural relationship to the isoprenoid‐type intermediates in the sterol pathway. These compounds were 1‐methyl‐4‐(1‐methyl‐ethenyl)‐cyclohexene, 6‐methyl‐5‐hepten‐2‐one, and 6,10‐dimethyl‐5,9‐undecadien‐2‐one. Hydroxyacetone was also found in all samples screened. The relationship of these compounds to the non‐sterol pathway of mevalonate metabolism is discussed.
Evaluation of vaginal malodor and efficacy of treatment by high performance ion exchange chromatography
Authors: Stanek Ronald , Glover D. D. , Mya Sandra , Larsen Bryan
Published: May 2005

High performance ion exchange chromatography was employed to evaluate the presence of short chain organic acids in the vaginal fluid of a woman troubled by persistent foul vaginal odor, but who did not have typical bacterial vaginosis. The vaginal secretions from this patient were collected on a weighed cotton swab and eluted into water and extracted by acidified ether. Salts of the acids were back‐extracted into aqueous solution and chromatographed on an H‐form resin column and compared to commercially available standards. A strikingly large amount of caproic acid was found. The caproic acid disappeared after metronidazole therapy, and a subsequent follow‐up chromatogram showed a predominance of lactic acid. The success of this technique in evaluating the present case suggests that such a method may prove useful in other types of vaginal infection.
Computerized chromatographic peak detection using the trigg tracking signal. An application devised for use with an online analog to digital converter connected between an amino acid analyser and a personal computer
Authors: Hartley T. F. , Philcox J. C. , Beesley J. F. , Robson J. B.
Published: May 2005

Manual integration of the amino acid peaks from physiological samples produced by conventional anion exchange liquid chromatography is a time‐consuming process. This paper describes a combined unit, consisting of an analog to digital converter and a personal computer, which was connected in parallel with the chart recorder and the analyser's 570 nm channel of the colorimeter. The computer was programmed to log the digitized data, detect the start, maximum and end of each chromatographic peak, calculate the area under the peak and its retention time and provide a printoct of the results at the end of the elution program. The computer program successfully exploited the Trigg Tracking Signal as both a peak detection and as a moving baseline monitoring device. This approach proved to have an equivalent performance to the manual method for 17 out of the 19 amino acids normally quantitated in physiological samples. The automated detection and quantitation of arginine was unsatisfactory due to its characteristically low profile peak shape, and proline was not measured because the device was not connected to the 440 nm channel of the colorimeter. The automated system provided economic and analytically acceptable solutions to the problem of providing an online integrator versatile enough to be used with the 255 min long amino acid analysis of physiological fluids.
Simultaneous liquid chromatographic determination of vanillylmandelic acid, homovanillic acid, and 5‐hydroxy‐3‐indoleacetic acid in urine, using isocratic elution and electrochemical detection
Authors: Richards D. A. , Titheradge A. C.
Published: May 2005

A method for the simultaneous measurement of vanillylmandelic acid, homovanillic acid and 5‐hydroxyindoleacetic acid in urine is described. Based on reversed‐phase liquid chromatography with electrochemical detection, the procedure employs isocratic elution, thus making it suitable for use in the less well‐equipped clinical or research laboratory. A simple extraction of the acids from acidified urine into ethyl acetate, is followed by evaporating to dryness a portion of the organic layer, and redissolving the residue in chromatographic mobile phase. Up to 20 samples can be analysed in a single working day. The method is validated and the results obtained are compared with refereènce methods. The cause of contamination of the glassy carbon surface of the working electrode is investigated, and a simple electrochemical pretreatment is described that overcomes this problem. Finally, the extra clinical information that can be derived from multi‐metabolite assays is considered.
Studies on the chromatographic fractionation of metabolites of benzo[a]pyrene in faeces and urine from germfree and conventional rats
Authors: Egestad Börje , Pettersson Per , Sjövall Jan , Rafter Joseph , Hyvönen Kaija , Gustafsson Jan‐Åke
Published: May 2005

Rats, germfree and conventional, were dosed with 14C‐labelled benzo[a]pyrene. Faeces and urine were collected. Metabolites in faces were effectively extracted with a new method using a combination of solvents and solid sorbents. Metabolites in urine were extracted with octadecylsilane‐bonded silica. The metabolites were fractionated into groups by chromatography on a cation exchanger (SP‐LH‐20 or SP‐Sephadex C‐25) and an anion exchanger (TEAP‐LH‐20). Some of the groups were further purified by column chromatography and analysed by HPLC and TLC. The analyses show a complex pattern of metabolism. A large part of the metabolites (9–24% depending on animal type and route of excretion) had amphoteric properties, e.g. like glutathione and cysteine conjugates. The abundance of conjugates sensitive to β‐glucuronidase and sulphatase was low. The relative amount of acidic conjugates in faeces was much higher in the germfree than in the conventional rats indicating the influence of the intestinal flora on the metabolism. The results support the view that the mercapturic acid pathway is a quantitatively important metabolic route for benzo[a]pyrene in rats. The methods of extraction and group fractionation were designed to be generally applicable to the analysis of lipophilic xenobiotics and their metabolites.
Rapid and simple HPLC determination of 5,5‐dimethyl‐2,4‐oxazolidynedione in rabbit serum
Authors: Gaździk Wiesław R. , Filipek Marian , Kmiotek Witold
Published: May 2005

A new method for the determination of 5,5‐dimethyl‐2,4‐oxazolidynedione (DMO) in rabbit serum by reversed phase‐HPLC with UV detection is described. The determination of DMO is performed without derivatisation. The internal standard is 5,5‐diethylbarbituric acid (barbital). The method is rapid and simple with sensitivity limit of 20 ng/mL, high recovery (above 92%) and is suitable for pharmacokinetic studies.
Study of doxorubicin photodegradation in plasma, urine and cell culture medium by HPLC
Authors: Le Bot M. A. , Riche C. , Guedes Y. , Kernaleguen D. , Simon S. , Begue J. M. , Berthou F.
Published: May 2005

An HPLC method was used to monitor the chemical stability of doxorubicin‐HCI (DXR) to light in plasma, urine and cell culture medium at room temperature. The results indicated that DXR was very unstable in cell culture medium and urine when exposed to light. It was more stable in plasma under the same conditions. In all the cases, the decrease in the amount of DXR is greatly dependent on light intensity (no noticeable degradation was observed after 8 h in the dark). These observations may be important for the correct interpretation of the effects and the toxicity of doxorubicin on cells incubated in cell medium, and for determination of urinary or plasma pharmacokinetic parameters.
Conditions for Quantitation of Dolichyl Phosphate, Dolichol, Ubiquinone and Cholesterol by HPLC
Authors: Elmberger P. Göran , Eggens Ivan , Dallner Gustav
Published: May 2005

Conditions for the isolation and quantitation of dolichyl phosphate, dolichol, cholesterol, and ubiquinone by reversed phase high performance liquid chromatography were investigated. A simple and fast sample preparation procedure using prepacked mini columns was employed. The UV spectra of the fractions obtained were examined and, in the case of dolichol compounds, the maximum absorbance around 205 nm was shown to be linearly dependent on the number of double bonds present in the isoprenolog. The analytical procedure described shows a very broad range of linearity (five orders of magnitude) and detects single dolichyl phosphate isoprenologs in amounts as small as 0.1 ng. The lowest overall recovery, that for dolichyl phosphate, is 77%. Use of isoprenolog 23 and ergosterol as internal standards reduced the variation in the method to 2.5, 4.0 and 5.5% for cholesterol, dolichyl phosphate and dolichol, respectively. The method described was employed to study the lipid composition of rat organs and biological variations in these compositions.
Separation of some antihistamines on impregnated TLC silica plates
Authors: Bhushan R. , Reena Chauhan  , Chauhan R. S.
Published: May 2005

Different metal salts have been tried as impregnating reagents for developing TLC separation schemes for some antihistamines on silica gel ‘G’ plates, using a new solvent system benzene + dimethyl formamide + acetic acid (30:10:7). Spots were visualized by spraying with a solution of Dragendroff's reagent.
Reversed phase HPLC of different membrane‐bound variant surface glycoprotein preparations from Trypanosoma brucei brucei
Authors: Tetaert Daniel , Hublart Marylègne , Gomès Véronique , Boersma Arnold , Degand Pierre
Published: May 2005

The chromatographic behaviour of the membrane‐attached variant surface glycoprotein (mfVSG) of bloodstream forms of Trypanosoma brucei brucei AnTat 1.1 A preparations were studied by reversed phase high performance liquid chromatography (RP‐HPLC). Among the different preparation procedures used, only the trifluoroacetic acid extraction gave a mfVSG preparation which was eluted from the RP‐HPLC column. As well as for the soluble variant surface glycoprotein (sVSG) it was found that different mfVSG forms were eluted at different organic solvent concentrations from the RP‐HPLC column. In addition, in preliminary studies, we have attempted to characterize the factors responsible for the trypanosome surface coat assembly. Using chromatographic techniques (RP‐HPLC and thin layer chromatography), the results suggest a close relationship between the content of lipid‐containing species and the heterogeneity of mfVSG preparation.
Determination of halofuginone in bovine plasma by competing‐ion high performance liquid chromatography after solid phase extraction
Authors: Kinabo L. D. B. , McKellar Q. A. , Murray M.
Published: May 2005

A high performance liquid chromatographic (HPLC) method for the determination of the anticoccidial and antitheilerial drug halofuginone in bovine plasma was developed. Samples were diluted with acetic acid (10%, v/v) and cleaned up on a Bond Elut C8 column. The analyte was eluted from the extraction column and chromatographed by reversed‐phase HPLC using decylamine as a competing‐ion reagent. Detection was by UV at 243 nm. Recovery from plasma was 75%, and within‐day and between‐day coefficients of variation were 5.23 and 6.35% respectively. The specificity and sensitivity of this method (limit of detection in plasma, 1 ng/mL) were sufficiently high to enable us to characterize the time course of the drug in plasma after oral administration of therapeutic doses to cattle.
High‐performance liquid chromatographic determination of β‐adrenoceptor blocking agents in body fluids
Authors: Šoltés Ladislav
Published: May 2005

Several reports have been published reviewing high‐performance liquid chromatographic (HPLC) methods for the determination of β‐adrenoceptor blocking agents (β‐blockers) in biological materials (Flouvat et al., 1981; Mehta, 1983; Marko and Šoltés, 1984; Ahnoff et al., 1985; Tkaczyková and Šafařík, 1987). Of these, the paper by Mehta (1983) briefly summarizes the interrelationship between physicochemical properties of β‐blockers with prechromatographic treatment of biological samples, as well as with the HPLC methods used for the determination of 12 β‐adrenoceptor blocking drugs. The work by Ahnoff et al. (1985) concerning the monitoring of cardiovascular drugs also deals with HPLC assays of 18 β‐blockers in plasma.

The Appendix to this report presents the great majority of HPLC methods for determining 30 β‐blockers in various body fluids. HPLC methods providing resolution and determination of individual β‐blocker enantiomers have not been included since this topic is being covered by Walle and Walle (1989). The Appendix is just a guide to the methods reviewed for the HPLC determination of parent β‐blockers as well as some of their metabolites co‐assayed in various body fluids. It does not include details such as the internal standard, recovery, setting of the detector, limit of determination, etc., given in the individual methods listed. The isolation technique of the drug(s) from the given body fluid represents the main step in the sample work‐up procedure. Along with this information, only the type of the HPLC column packing and the detection principle used by each method's developers are given.
High performance liquid chromatographic analysis of time‐dependent changes in urinary excretion of indoleamines following tryptophan administration
Authors: Tsuchiya Hironori , Ohtani Shigeru , Takagi Nobuhiko , Hayashi Tokishi
Published: May 2005

Analytical conditions of prepurification extraction and HPLC separation were optimized for determination of urinary serotonin and tryptamine. Under optimal conditions, serotonin, tryptamine and an internal standard were extracted with 15% v/v n‐propanol in diethyl ether from urine samples alkalized with a phosphate buffer (0.75 mol/L, pH 10.0), and then they were re‐extracted into an HCI solution (0.1 mol/L). Purified indoleamines were simultaneously separated by reversed‐phase ion‐pair HPLC with native fluorescence detection. Urinary serotonin and tryptamine were selectively determined within about 45 min per sample for the whole procedure. Analytical recovery, reproducibility and detection sensitivity were satisfactory for pursuing time‐dependent changes in indoleamine levels. Urinary excretion profiles of serotonin and tryptamine in subjects dosed with L‐tryptophan were successfully analyzed by our method.
Determination of total and free concentration of propranolol in human plasma by displacement electrophoresis in a two‐layer polyacrylamide gel using fluorimetric detection
Authors: Zelikman Ilya , Hjertén Stellan
Published: May 2005

A new method based on displacement electrophoresis has been developed for the determination of the total and free concentration of propranolol, a β‐adrenergic blocker drug, in plasma. To determine the total concentration the drug is extracted from human plasma into a chloroform + heptane mixture in the presence of ammonia. After evaporation of the solvent mixture the residue is submitted to displacement electrophoresis in a glass tube containing a two‐layer polyacrylamide gel. When the propranolol electrophoretically leaves the gel column it is transferred by a buffer flow to the cuvette of a fluorimeter for continuous detection and quantification. The concentration of the free non‐protein bound drug can be determined by the same displacement electrophoresis technique following extraction of the plasma sample into the above organic solvent mixture in the absence of ammonia. Alternatively the extraction procedure can be exchanged for dialysis for 1 h. To decrease the risk that large size material might comigrate with propranolol two layers of polyacrylamide are used, one having small pores to retard such material. In addition, we use fluorimetric detection which means that many possible contaminants are not recorded and therefore do not affect a quantitative determination of propranolol.