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PubMed2004

Utility of endogenous creatinine clearance as a measure of renal function in mice.

BACKGROUND: The use of endogenous plasma creatinine levels and creatinine clearance as a tool to evaluate renal function in mice has come under scrutiny as prior studies have reported that the Jaffé alkaline picrate method grossly overestimates true plasma creatinine in mice. As members of the NIDDK Animal Models of Diabetic Complications Consortium (AMDCC), we evaluated the performance and feasibility of an alternative high-performance liquid chromatography (HPLC)-based method for standard determination of plasma creatinine and creatinine clearance in mice. Our purpose was to develop a simple method that provides a reliable, reproducible, and sensitive assay for small volumes (<25 microL) of mouse plasma and sera. METHODS: We compared creatinine clearance measured by HPLC with the Jaffé method and HPLC creatinine clearance with inulin clearance [fluoroscein isothiocyanate (FITC) inulin in an osmotic pump implanted in mouse] in C57BL/6J mice. Different groups of mice underwent either one of two protocols. Protocol A included dietary intervention with normal, low salt plus enalapril, or high salt. Protocol B induced diabetes using streptozotocin. RESULTS: First, mean plasma creatinine levels were significantly lower (P < 0.0001) by HPLC (0.128 +/- 0.026 mg/dL) vs. Jaffé (0.4 +/- 0.12 mg/dL) for mice on a normal diet. Urine creatinine concentrations measured by HPLC were 10% lower than by Jaffé (P < 0.01). Second, mean creatinine clearance by HPLC for mice on a normal diet was 255 +/- 68 microL/min. Mice on low salt diet plus enalapril had reduced creatinine clearance (72.8 +/- 24.2 microL/min) while mice on high salt diet had an elevated creatinine clearance (355 +/- 105 microL/min). Third, diabetic mice (19 to 24 weeks of diabetes) exhibited hyperfiltration as creatinine clearance was 524 +/- 214 microL/min whereas nondiabetic age/gender-matched mice showed a mean creatinine clearance of 206 +/- 41 microL/min. Finally, significant correlation was demonstrated for creatinine clearance by HPLC vs. inulin clearance (R= 0.643; P < 0.001). CONCLUSION: HPLC is highly accurate, much more sensitive and specific than the Jaffé method for plasma creatinine measurements in mice. Creatinine clearance in mice measured by HPLC reflects changes in renal function induced by diet and diabetes.

Animals↗
PubMed1980

An improved continuous flow method for serum creatinine using the Jaffé reaction.

A method for the measurement of creatinine is described in which serum is dialysed directly into a combined alkaline picrate reagent. Baseline noise and drift are eliminated, and precision is improved by avoiding the need to add further reagents after dialysis. This may be of particular interest to laboratories that perform this assay on first-generation AutoAnalyzer equipment.

Autoanalysis↗
PubMed1987

Investigations into negative interference by jaundiced plasma in kinetic Jaffé methods for plasma creatinine determination.

Negative interference by jaundiced plasma in kinetic Jaffé methods for plasma creatinine determination is caused by both conjugated and unconjugated bilirubin. The amount of interference observed is method- and temperature-dependent and is directly related to bilirubin concentration but is independent of the creatinine concentration. Bilirubin also significantly reduces the precision of the creatinine estimation. A mechanism of bilirubin interference in kinetic Jaffé methods is proposed and the influence of picrate concentration, hydroxide concentration and temperature on the magnitude of bilirubin interference has been investigated.

Bilirubin↗
PubMed1982

Studies on chemotherapy of parasitic helminths (VI) effects of various neuropharmacological agents on the motility of Dipylidium caninum.

Effects of various neuropharmacological agents on the motility of Dipylidium caninum was studied. 5-HT stimulated the motility of D. caninum, while other drugs such as GABA, adrenaline, phenylephrine, isoproterenol, dibenamine, and propranolol showed little effect. The stimulatory action of 5-HT was antagonized by tryptophol. Stibnal caused paralysis which was blocked by the pretreatment with 5-HT. Paralytic effects were caused by eserine, dichlorovos, ACh, carbachol and DMPP, but not by pilocarpine and McN-A-343. d-Tubocurarine, hexamethonium, and atropine all showed little effect on the motility and on the paralytic action of eserine. Though guanidine showed little effect, strychnine remarkably stimulated the motility. The action of strychnine was stimulated by 5-HT but partially inhibited by tryptophol. Morphine and picrate slightly stimulated the motility, but showed little influence on the paralytic action of eserine. These two agents stimulated the paralyzed preparation by tryptophol transiently and sustainedly, respectively. These results on the cholinergic and serotonergic drugs basically showed good agreement with those reported in trematodes such as Schistosoma mansoni and Fasciola hepatica.

Animals↗
PubMed1987

Specific creatinine determination in laboratory animals using the new enzymatic test kit "Creatinine-PAP".

The new Creatinine-PAP test kit from Boehringer Mannheim GmbH was evaluated for the determination of creatinine in dogs, rabbits, guinea pigs, rats, and mice. In comparison with methods based on the reaction with alkaline picrate, the enzymatic method gave the lowest serum creatinine values. The differences are caused by serum substances (non-creatinine chromogens) reacting with picric acid. Noticeable differences between the standard inulin clearance and creatinine clearance (e. g., in rats) arise because the non-creatinine chromogens interfere differently in the picric acid methods, and serum and urine contain different concentrations of interfering substances. The concentrations of non-creatinine chromogens are different in the various laboratory animals. Since these substances cannot be entirely removed (e. g., in rats, dogs) by the adsorption procedure with Fuller's earth, we recommend the peroxidase-coupled enzymatic test as a practical and specific method for routine measurements of creatinine in laboratory animals.

Animals↗
PubMed2001

Selective transport of silver ion through a supported liquid membrane using hexathia-18-crown-6 as carrier.

A facile supported liquid membrane (SLM) system for the selective and efficient transport of silver ion is introduced. The SLM used is a thin porous polyvinyldifluoride membrane impregnated with hexathia-18-crown-6 (HT18C6) dissolved in nitrophenyloctyl ether. HT18C6 acts as a specific carrier for the uphill transport of Ag+ ion as its picrate ion paired complex through the SLM. In the presence of thiosulfate ion as a suitable stripping agent in the strip solution, transport of silver occurs almost quantitatively after 4 h. The selectivity and efficiency of silver transport from aqueous solutions containing other Mn+ cations such as Mg2+, Ca2+, Co2+, Ni2+, Cu2+, Zn2+, Pb2+, Cd2+, Hg2+, Fe3+ and Cr3+ ions were investigated.

Chemical Phenomena↗
PubMed1988

Evaluation of several creatinine methods in search of a suitable secondary reference method: report from the subcommittee on reference method for creatinine, Nordic Society for Clinical Chemistry.

Several methods for creatinine have been examined by the subcommittee on reference method for creatinine. These include the use of picric acid to precipitate proteins followed by alkalization, or the use of alkaline picrate after protein precipitation and adsorption to Fuller's earth. The first approach was not found to be suitable as a reference method since it was susceptible to matrix effects, fluctuations in temperature and wavelengths. The second approach was less sensitive to these effects but showed variations with different batches of Fuller's earth. The method using high performance liquid chromatography (HPLC) had poor precision, probably because endogenous compounds co-eluted either with creatinine or with the internal standard. The enzymatic method based on the determination of hydrogen peroxide had good accuracy and precision. However, the main disadvantage is that the method is sold as a kit and detailed specifications concerning all the ingredients are not available to permit independent studies using the basic components.

Chromatography, High Pressure Liquid↗
PubMed1992

Oxidation markedly reduces bilirubin interference in the Jaffé creatinine assay.

Bilirubin causes underestimation of serum creatinine in the Jaffé alkaline picrate assay. We report an approach for preventing bilirubin interference by pretreating serum samples with peroxidase and H2O2. The dissociation of bilirubin from albumin and its subsequent oxidation markedly reduces the bilirubin interference and enables accurate determination of creatinine concentrations by the Jaffé reaction even in hyperbilirubinemic sera. Within-run CVs were 2.6%, 4.0%, and 3.8% at mean creatinine concentrations of 88, 165, and 349 mumol/L, respectively (n = 20). Day-to-day CVs were 4.0%, 6.3%, and 5.8% for mean creatinine concentrations of 87, 168, and 364 mumol/L, respectively (n = 12). Average recovery of creatinine added to serum in the presence of 600 mumol/L bilirubin was 97% (n = 15). This method requires only small serum volumes (70 microL) and is easily applicable to automated analyzers that can be programmed to add three reagents consecutively.

Bilirubin↗
PubMed1989

Nitromethane interferes in assay of creatinine by the Jaffé reaction.

A patient who ingested a methanol/nitromethane mixture (model airplane fuel) showed an apparent plasma creatinine concentration of 8.0 mmol/L by the Jaffé reaction when the actual creatinine concentration, as measured by a specific enzymatic method, was 0.09 mmol/L. This effect was due to nitromethane in the plasma. Interference by nitromethane with the determination of creatinine by reaction with alkaline picrate (the Jaffé reaction) has not previously been reported. When nitromethane was added to plasma, the apparent creatinine measured was linearly related to the amount of nitromethane added. Comparison of spectral changes occurring during creatinine/picrate and nitromethane/picrate reactions show substantial similarity, suggesting a similar structure for the products. Although the findings are of interest for both their toxicological and analytical implications, the main interest lies with their suggestion of a model system for future investigation of the Jaffé reaction.

Adult↗
PubMed1987

Interference by the new antibiotic cefpirome and other cephalosporins in clinical laboratory tests, with special regard to the "Jaffé" reaction.

The new cephalosporin, cefpirome, was investigated for its possible interference in clinical laboratory tests, especially the determination of creatinine. Tests for bilirubin, cholesterol, protein, urea, and uric acid were also studied. A selection of commercially available compounds such as cefoperazone, cefazolin, cefamandole, cefoxitin, latamoxef, ceftriaxon, cefotiam, cephaloridine, cephalotin, cefotaxime, cefazedone and cefuroxime, and cefodizime (a compound under development) were also included in these investigations. Interference was found only with the "Jaffé method" for the determination of creatinine. Pronounced interactions with alkaline picrate were observed with cefpirome, cefoxitin, cephalotin and cephaloridine, whereas the other compounds showed only weak reaction or no reaction at all. The cephalosporins did not interfere in any of the other tests mentioned above. Care must therefore be taken in the determination of creatinine in samples from patients under treatment with these drugs. The assay methods of choice at present are enzymatic or specific HPLC assays.

Blood Chemical Analysis↗
PubMed1977

Positive interference with the Jaffé reaction by cephalosporin antibiotics.

Cephaloglycin, cephalothin, cephaloridine, cefoxitin, and cephacetrile are cephalosporin antibiotics that react with alkaline picrate solution to give a creatinine-like response. The mechanism of this reaction is not known, and several structurally similar compounds do not react under these same conditions. Because large quantities of a cephalosporin antibiotic may be administered to a patient with limited ability to excrete the drug (as occurs with compromised renal function), the magnitude of this interference can be appreciable. It does not appear possible to avoid the cephalosporin-caused interference by the use of "kinetic" creatinine methods.

Cephalosporins↗
PubMed1996

[Method selected for the determination of creatinine in plasma or serum. Choice of optimal conditions of measurement].

The method selected by the SFBC (Société française de biologie clinique) is derived from the colorimetric reaction of creatinine with alkaline picrate, measured kinetically, without any pretreatment step. The key parameters of the reaction determining the quality of the results are studied, with special regard to samples including known interferents. The aims of the study were to gain an optimal analytical sensitivity and to reduce main interferences (acetoacetate, bilirubine, glucose, protein) which plague the Jaffé reaction, through a comprehensive study of the reagents, of their concentrations and of the analytical procedures. The selected concentrations (in the test) are: 150 mmol/L sodium hydroxide, 10 mmol/L picric acid and 2 g/L sodium dodecyl sulfate. Ten millilitres of a BRIJ solution (30% volvol) are added to the reagent. The operating procedures are as follow: sample ratio 0.07 to 0.08; wavelength 505 to 510 nm; temperature 37 degrees C; incubation of the specimen with the alkaline reagent 5 mn (at least), before starting the reaction with picric acid. A seric calibrator is recommended. The first measurement is taken 20 to 40 s after starting the reaction. Total measurement time is 120 to 150 seconds.

Blood Chemical Analysis↗
PubMed1991

Oxoaporphine alkaloids: conversion of lysicamine into liriodendronine and its 2-O-methyl ether, and antifungal activity.

Pschorr reaction of diazonium salt 7 in aqueous methanolic sulfuric acid afforded, besides lysicamine 2, the orange colored sulfate of oxodibenzopyrrocoline (8). The structure is fully supported by an X-ray analysis of its picrate salt. Selective ether cleavage of lysicamine (2) with 48% HBr afforded a hydrobromide of 9, and free betaine 9 on treatment with pyridine-water. Both compounds methylated on treatment with etherial diazomethane on nitrogen to give the known 2-O,N-dimethylliriodendronine (11). Liriodendronine (10) was obtained from lysicamine (2) on heating with pyridine HBr at 189 degrees C, and treatment with pyridine-water, as a dark violet betaine. Betaine 12 was obtained by heating 11.HCl to 200 degrees C. The quaternary salts of lysicamine, lysicamine methiodide (3) and lysicamine methosulfate (4) were comparable in anticandidal activity to liriodenine (1), but were not as active as liriodenine methiodide (13).

Alkaloids↗
PubMed2005

A new type of migrating zone boundary in electrophoresis: 1. General description of boundary behavior based on electromigration dispersion velocity profiles.

Till now two principal types of electrophoretic boundaries have been recognized: a steady-state one showing self-sharpening properties and an unsteady-state one showing electromigration dispersion (EMD). In this contribution, the existence of a third (hybrid) type of electrophoretic boundary is revealed, that shows simultaneously: (i) steady-state character with self-sharpening properties in a certain part of the boundary, demarcated by a certain range of its composition; (ii) unsteady-state character with EMD in the resting part. A new theoretical approach to the study of the structure and properties of electrophoretic boundaries is presented, based on EMD velocity profiles representing the dependence of the EMD velocity on the composition in any point of the transition region across the boundary. According to this approach, the linearity or nonlinearity of the EMD velocity profile is recommended as the criterion for distinguishing the actual type of the boundary in question. It is shown that the new type of electrophoretic boundary is not an exotic case but may be observed even in simple systems as, e.g., formed by adjacent zones containing mixtures of current species such as picrate and acetate. Computer simulations are presented which confirm the theoretical conclusions and illustrate the three types of electrophoretic boundaries.

Electrophoresis, Capillary↗
PubMed1990

High resolution scanning electron microscopy of stereocilia in the cochlea of normal, postmortem, and drug-treated guinea pigs.

The morphology of hair bundles has been studied by high resolution scanning electron microscopy using a variety of fixatives, including glutaraldehyde, glutaraldehyde-picrate, glutaraldehyde-tannic acid, glutaraldehyde followed by post-fixation in osmium tetroxide, and the osmium thiocarbohydrazide technique. Critical evaluation of several metal coatings, gold, gold-palladium, and platinum has been carried out. Both the surface texture of stereocilia and their cross-links are sensitive to fixation and metal coating. We are of the opinion that glutaraldehyde gives the best general quality of fixation and preservation for all types of cross-links. We have described three major sets of cross-links: first, lateral links connecting stereocilia within the same row; second, lateral links connecting stereocilia of adjacent rows; and third, upward-pointing links, one per stereocilium, connecting the tip of each shorter stereocilium to the lateral surface of the adjacent taller stereocilium. Current physiological and anatomical evidence suggests that the lateral links couple the individual stereocilia within the hair bundle so that they function as a single mechanical unit. The upward-pointing tip links are ideally placed to respond to mechanical deformation of the hair bundle, being stretched when the stereocilia are deflected in the excitatory direction towards the tallest row and relaxed when deflected in the opposite, inhibitory direction. Postmortem morphological changes are detected within 15 minutes of cardiac arrest and become progressively more pronounced in time. These results enabled us to distinguish specific drug-induced changes which could not be attributed simply to cell death. Effects of cisplatin and kanamycin upon hair bundles are described. The work reported here is based on studies using the guinea pig cochlea. Some of the postmortem changes described have also been confirmed in human cochleas. It is stressed that many of the postmortem and drug-induced effects can only reliably be studied by high resolution scanning electron microscopy coupled with appropriate preparation procedures.

Animals↗
PubMed1977

Analysis of homatropine methylbromide dosage forms.

A stability-indicating method of analysis of homatropine methylbromide in pharmaceutical formulations was developed. This method is based on the formation of a picric acid-quaternary ammonium complex, which is adsorbed on acid-washed diatomaceous earth in alkaline media followed by on-column chloroform extraction. The picrate complex is measured spectrophotometrically at 365 nm. The method was selective for homatropine methylbromide in that there was no interference from its major hydrolytic decomposition products, tropinium methylbromide and mandelic acid.

Methods↗
PubMed1984

Ferricenium complexes: a new type of water-soluble antitumor agent.

The antitumor activity of a series of iron complexes, i.e., of ferrocene [Cp2Fe], of tetrachloroferrates(III) [R4N]+[FeCl4]-, and of ferricenium complexes [Cp2Fe]+X- (X- = [FeCl4]-, 1/2 [Cl3FeOFeCl3]2-, [H5Mo7O24]- X 2H2O, [2,4,6-(NO2)3C6H2O]-, or [CCl3COO]- X 2 CCl3COOH) was investigated against EAT in CF1 mice. Whereas ferrocene and the ammonium tetrachloroferrates(III) did not show recognizable tumor-inhibiting activity, such activity was exhibited by the water-soluble, salt-like ferricenium complexes; the best antineoplastic properties, with optimum cure rates of 100%, were found for ferricenium picrate and ferricenium trichloroacetate. The ferricenium compounds are the first iron complexes for which antineoplastic activity has now been shown. They represent a new type of antitumor agent insofar as they differ fundamentally from known inorganic and organometallic antitumor agents (a) by their ionic, salt-like character, which is responsible for their high water solubility, and (b) by the absence of a cis-dihalometal moiety; this moiety has been recognized as important for the intracellular action of other known inorganic cytostatics.

Animals↗