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PubMed2001

Lactone formation of N-acetylneuraminic acid oligomers and polymers as examined by capillary electrophoresis.

We examined lactone-formation reaction of oligomers and polymers of N-acetylneuraminic acid in diluted hydrochloric acid solution and found that the time course of the lactonization reaction was easily traced by capillary electrophoresis. The reaction proceeded more rapidly with increasing the molecular mass of oligomers because the conformation of inner units became rigid and more favorable for the formation of lactone linkage. Present results obtained using capillary electrophoresis will be useful in understanding of physical and chemical properties of oligo/polysialic acids.

Carbohydrate Sequence↗
PubMedDate not supplied

Measurement of N-acetylneuraminic acid in human serum and urine by high performance liquid chromatography with chemiluminescence detection.

A highly sensitive method for the determination of N-acetylneuraminic acid in human serum and urine is investigated. This method employs high performance liquid chromatography with chemiluminescence detection. N-Acetylneuraminic acid, released by hydrochloric acid hydrolysis of serum and urine, and N-glycolylneuraminic acid (internal standard) are converted into chemiluminescent derivatives with 4,5-diaminophthalhydrazide dihydrochloride, a chemiluminescence derivatization reagent for alpha-keto acids. The derivatives are separated within 35 min on a reversed phase column, TSKgel ODS-120T, with isocratic elution, followed by chemiluminescence detection; the chemiluminescence is produced by the reaction of the derivatives with hydrogen peroxide in the presence of potassium hexacyanoferrate(III) in alkaline solution. The detection limit for N-acetylneuraminic acid is 9 fmol (signal-to-noise ratio = 3). This sensitivity permits precise determination of N-acetylneuraminic acid in 10 nL of serum or 50 nL of urine. The method is applied to the determination of the N-acetylneuraminic acid in human sera from normal subjects and cancer patients and in normal urine.

Adult↗
PubMed1994

Elutions of metal ions from dental casting alloys and their effect on calcium phosphate precipitation and transformation.

The elution characteristics of metal ions from dental casting alloys were studied in vitro. Large amounts of nickel (0.12-4.94 mg/cm2) and chromium (< 0.01-0.63 mg/cm2) were solubilized from the surface of seven brands of nickel-chromium alloy in either 1% lactic acid or 0.05% hydrochloric acid solutions. The elution of chromium from two brands of cobalt-chromium alloys in both eluents was below the detection limit or less than 0.01 mg/cm2. The elution of tin, copper, and zinc from a gold-palladium-silver alloy in both eluents was below the detection limit or less than 0.01 and 0.04 mg/cm2. Some amounts of tin (0.19-1.92 mg/cm2) and zinc (0.56-1.73 mg/cm2) were eluted from a silver alloy in both eluents. The effects of five eluting metal ions, i.e., nickel, chromium, tin, copper, and zinc, on the conversion of amorphous calcium phosphate (ACP) to hydroxyapatite (HAP) in vitro also were studied by a pH drop method. All divalent cations except for chromium decreased the rate of HAP transformation and elongated the induction time. Nickel had an inhibitory effect comparable to 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) on the rate of HAP transformation. Tin, copper, and zinc inhibited similarly, but the inhibition was weaker than that by nickel. Chromium did not inhibit these reactions.

Calcium Phosphates↗
PubMed1993

31P NMR examination of phosphorus metabolites in the aqueous, acidic, and organic extracts of Phaseolus vulgaris seeds.

31P NMR spectroscopy was used to compare the phosphorus compound content in aqueous, acidic, and organic extracts of Phaseolus vulgaris seeds. Two chloroform-methanol and three ethanol extractions were used to isolate phospholipids from dry cotyledons and the phospholipid profiles were compared. Variations in phospholipid composition and artificially high concentrations of lysophosphatidylcholine were observed in the ethanol extracts. Other phosphorus compounds were extracted using perchloric acid, trichloroacetic acid, trichloroacetic acid in ether, hydrochloric acid, boiling water, and aqueous Hepes. The Hepes extraction was introduced in order to compare the results of a gentle procedure with those involving heat and acid treatments. Low concentrations of phospho-sugars and other phosphorus metabolites were found in all these extracts. High concentrations of phytate were found in all aqueous and acidic extractions; however, 31P NMR spectra of aqueous extractions did not show the phytate resonance. The NMR silence of the phytate resonances is attributed to complexation of phytate. The 31P NMR spectra of aqueous extracts contained a broad resonance at 0.1 ppm assigned to protein-bound RNA. Various extraction procedures on the same biological material are presented, and the comparison of these methods will facilitate future analysis and interpretation of literature reports involving these methods of tissue analysis.

Chemistry Techniques, Analytical↗
PubMed1981

[Vagal acido- and glucoreceptors in the gastro-duodenal region (author's transl)].

Vagal sensitivity to carbohydrates (mainly glucose) and to acids (hydrochloric and acetic acids) was studied in the gastro-duodenal region of anesthetized cats. Action potentials were recorded extracellularly from the nodose ganglion by means of glass microelectrodes. Receptors responding to glucose perfusion were found at this level as well as receptors stimulated by acid perfusion. It is shown that each type of receptor was activated by only one kind of stimulus (carbohydrates and acids, respectively). These receptors must thus be considered as true glucoreceptors or acido-receptors. Being silent before activation, these two types of chemoreceptors discharged at a frequency that varied from 2 to 14 imp/s for the acido-receptors, and from 2 to 30 imp/s for the glucoreceptors. The response could be irregular or regular and might last several minutes. Its latency was short (between 1 and 20 s for both types of chemoreceptors. It was therefore supposed that they were located in the mucosa or neighboring structures. From the conduction velocities (0.8-1.2 m/s), it was concluded that fibers originating from the acido- and glucoreceptors belonged to the C type. Recording of the electromyographic activity from the gastro-duodenal region demonstrated that the gluco- and acid-receptors are involved in the regulation of motility of the gastric and duodenal regions.

Action Potentials↗
PubMed1978

Glucose in vertebrate skeletal muscle proteins.

Neutral sugar was found in proteins extracted with acid chloroform/methanol from skeletal muscle of frog, normal and dystrophic chicken as well as in conventional preparations of frog tropomyosin and rabbit myosin. Whenever tested, the sugar content remained unaffected by two extractions with 5% trichloroacetic acid and by treatment with acid (0.1 N hydrochloric acid) or alkali (0.1 N sodium hydroxide, 60--90 min, 100 degrees C). These and other chemical properties favor the notion that sugar is present in covalent linkage to protein rather than as an attending contaminant. In every protein studied sugar profiles consisted of glucose only except rabbit myosin where it was accompanied by another sugar, presumably ribose.

Animals↗
PubMed1992

Astringent compounds suppress taste responses in gerbil.

Astringent tastes are generally considered those that induce long-lasting puckering and drying sensations on the tongue and membranes of the oral cavity. Electrophysiological recordings were made here from the whole chorda tympani nerve in gerbil to understand the interactive effect of astringent-tasting molecules with a broad spectrum of tastants including mono- and divalent salts, bitter compounds, acids, and sweeteners. The astringent tasting compounds were tannic acid (24 mM at pH's 2.9 and 5.5), aluminum ammonium sulfate (30 mM), aluminum potassium sulfate (10 mM) and gallic acid (30 mM). Hydrochloric acid (1 mM, pH 2.9) was also tested to control for acidity, since aqueous solutions of astringent-tasting compounds are acidic. Adaptation of the tongue to tannic acid (24 mM) at both pH 2.9 and 5.5 markedly inhibited responses elicited by salts, acids, sweeteners, and bitter-tasting compounds. The degree of the inhibition at these two pH values is about the same which suggests that tannic acid itself (as opposed to acidity) may produce this inhibition. Chorda tympani responses to sweeteners were completely suppressed by tannic acid; responses to KCl, NH4Cl, and urea were the least suppressed. The aluminum salts also inhibited the chorda tympani responses to all stimuli tested. Gallic acid, which is weakly astringent, had minimal effects on the chorda tympani responses to the test compounds. These data suggest that both tannic acid and the aluminum salts inhibit a variety of transport pathways and receptors in taste cells for a broad spectrum of tastants. The inhibition of some of these pathways may contribute to the astringent taste sensation.

Aluminum↗
PubMed1992

Studies on the methanolysis of small amounts of purified phospholipids for gas chromatographic analysis of fatty acid methyl esters.

The methanolysis of small amounts of purified phosphoglycerides and sphingomyelin was studied and a quantitative comparison of five methods for the methanolysis of standard phosphoglycerides was made. These methods were based on methanolysis with boron trifluoride-methanol, methanolic sodium methoxide (at ambient temperature and with heating) and methanolic sulphuric acid. A further method was based on saponification with methanolic sodium hydroxide and subsequent esterification with boron trifluoride-methanol. Under the experimental conditions, only the sodium methoxide-catalysed method at ambient temperature gave complete methanolysis of phosphoglycerides. For methanolysis of sphingomyelin, boron trifluoride-methanol, methanolic sulphuric acid and methanolic hydrochloric acid were used. It was found that complete methanolysis of sphingomyelin takes 15 h at 90 degrees C. Based on these results, procedures for the methanolysis of phosphoglycerides and sphingomyelin separated by high-performance liquid chromatography are presented.

Animals↗
PubMed1985

Cardiorespiratory changes following chemical applications to gut serosa.

Application of 0.2-0.3 ml of 1 M ammonium chloride, 100 mM citric acid, 100 mM hydrochloric acid, 1% potassium chloride, 1% ammonium oxalate, 1% oxalic acid, 0.5% sodium hydroxide, 200 micrograms% bradykinin or 20 mg% capsaicin to gastrointestinal serosa produced a fall in blood pressure and inhibition of respiration in hypertensive as well as normotensive rats. In 6 (8%) of the animals studied, a fall in heart rate was also seen which was blocked by atropine. The blood pressure changes were independent of changes in heart rate. After acute abdominal vagotomy, while the respiratory inhibition following chemical application on gut serosa was enhanced, the cardiovascular responses were the same as before. In animals subjected to sympathetic denervation by spinal section at T5, celiac ganglionectomy or splanchnicotomy, a similar application produced an augmented respiratory response while the cardiovascular parameters were unaffected. These results show that the above inhibitions were mediated through the sympathetic afferents and a minor stimulation of respiration, via vagal afferents. The efferent pathway for the cardiac changes seems to be via vagus and the vascular changes via the sympathetics. In chronic vagotomized animals, the same stimulation produced a response similar to that seen in animals with intact vagus. An adaptive response may be operating in chronic animals reverting the status back to normal.

Ammonium Chloride↗
PubMed1985

Determination of SQ 27,519, the active phosphinic acid-carboxylic acid of the prodrug SQ 28,555, in human serum by capillary gas chromatography with nitrogen-phosphorus detection after a two-step derivatization.

A method for the determination of SQ 27,519 (II), the active phosphinic acid-carboxylic acid of the prodrug SQ 28,555 (I), in human serum is presented. Compounds I and II are simultaneously extracted from acidified serum into ethyl acetate, and II is back-extracted into aqueous sodium bicarbonate. Compound I, in ethyl acetate, can be subsequently hydrolyzed and measured as II. The two acidic groups of II are selectively esterified, first by methylation of the carboxylic acid with methanolic hydrochloric acid and then by formation of the hexafluoroisopropyl ester of the phosphinic acid. The resulting product is measured by splitless-injection capillary gas chromatography with nitrogen-phosphorus detection. Linear standard curves were obtained for II with a detection limit of less than 10 ng/ml of serum. The method was successfully applied to the analysis of serum samples obtained from normal individuals after administration of I. In an ascending-dose study involving several human subjects the serum levels of II ranged from less than 10 to 7000 ng/ml of serum.

Chemical Phenomena↗
PubMed2005

Lead biosorption and desorption by intact and pretreated spirulina maxima biomass.

In order to search for locally available and untried biomaterials in China with high removal capacity of heavy metals from wastewater, the feasibility of Spirulina maxima as biosorbent for lead removal and recovery from aqueous solution was investigated. The lead biosorption was studied by using intact biomass and pretreated biomass of S. maxima. The effects of operational conditions (e.g. pH, contact time, biomass concentration etc.) on lead biosorption were investigated. The biosorption was solution pH dependent and the maximum adsorption was obtained at a solution pH of about 5.5. The adsorption equilibrium was reached in 60 min. The biosorption followed the Freundlich isotherm model. The maximum removal ratios of lead were about 84% in intact biomass and 92% in pretreated biomass. The lead adsorbed could be desorbed effectively by 0.1 M nitric acid, EDTA and hydrochloric acid. The results in this study indicated that pretreated biomass of S. maxima was a promising candidate for removing lead from wastewater.

Adsorption↗
PubMed2004

Liquid chromatographic method for the quantitative determination of Nepsilon-carboxymethyllysine in human plasma proteins.

The modification of the lysine moieties of proteins to Nepsilon-carboxymethyllysine (CML) is supposed to play a major role in the development of long-term complications in patients with diabetes mellitus. This paper presents an analytical method for the quantitative determination of CML in plasma proteins, which could be used for studying the development of diabetic complications. The method is based on isolating proteins from plasma by precipitation with trichloroacetic acid and hydrolysing these under acidic conditions (6M hydrochloric acid at 110 degrees C for 20 h) to the individual amino acids. After hydrolysis, CML is derivatised along with the other amino acids to 9-fluorenylmethoxycarbonyl (FMOC) derivatives, which are subsequently separated by reversed-phase column liquid chromatography using a 150 mm x 4.6 mm C8 column and a mobile phase of 25 mM potassium phosphate buffer (pH 2.0) and acetonitrile (80:20 (v/v)) and detected using fluorescence detection (excitation at 260 nm and emission at 310 nm). Quantification of the protein-bound CML content of a plasma sample is achieved using standard addition. The impact of several aspects of the sample preparation and chromatography on method performance is discussed. Method evaluation results are reported and show that this method is capable of determining CML with good accuracy and precision (below 10%) in the relevant concentration range (1-10 microg/ml), with a limit of detection of 0.2 microg/ml.

Blood Proteins↗
PubMed2005

A stability indicating LC method for rivastigmine hydrogen tartrate.

An isocratic, reversed-phase liquid chromatographic (RPLC) method was developed for the quantitative determination of Rivastigmine hydrogen tartrate, a cholinesterase inhibitor in bulk drugs and in pharmaceutical dosage forms. The developed method is also applicable for the related substance determination of Rivastigmine hydrogen tartrate in bulk drugs. The chromatographic separation was achieved on a Waters X Terra RP18 (250 mm x 4.6 mm, 5 microm) column using aqueous 0.01 M sodium-1-heptane sulphonate (pH: 3.0 with dilute phosphoric acid)-acetonitrile (72:28, v/v) as a mobile phase. The chromatographic resolution between Rivastigmine and its potential impurity, namely (S)-3-(1-dimethylaminoethyl) phenol (Imp 1) was found to be greater than four. Forced degradation studies were performed for Rivastigmine hydrogen tartrate bulk drug using acid (0.5 N hydrochloric acid), base (0.5 N sodium hydroxide), oxidation (3% hydrogen peroxide), heat (60 degrees C) and UV light (254 nm). No degradation was observed for Rivastigmine hydrogen tartrate except in base hydrolysis and the formed degradation product was found to be Imp 1. The mass balance of Rivastigmine hydrogen tartrate was close to 100 in all the stress conditions. The limit of detection (LOD) and limit of quantification (LOQ) of Imp 1 were found to be 100 and 300 ng/ml, respectively, for 10 microl injection volume. The percentage recovery of Imp 1 in bulk drug sample was ranged from 95.2 to 104.3. The active pharmaceutical ingredient was extracted from its finished dosage form (capsule) using water. The percentage recovery of Rivastigmine hydrogen tartrate was ranged from 99.2 to 101.3 and 98.6 to 101.5 in bulk and pharmaceutical formulation samples, respectively. Rivastigmine hydrogen tartrate sample solution and mobile phase were found to be stable for at least 48 h. The developed method was validated with respect to linearity, accuracy, precision, robustness and forced degradation studies prove the stability indicating power of the method.

Chromatography, Liquid↗
PubMed2006

Development and validation of a specific stability indicating high performance liquid chromatographic method for rizatriptan benzoate.

A gradient, reversed-phase liquid chromatographic (RP-LC) method was developed for the quantitative determination of rizatriptan benzoate, used to treat relieves migraine headache symptoms. The developed method can be also employed for the related substance determination in bulk samples. Forced degradation studies were performed on bulk sample of rizatriptan benzoate using acid (0.5 N hydrochloric acid), base (0.1 N sodium hydroxide), oxidation (3.0% hydrogen peroxide), water hydrolysis, heat (60 degrees C) and photolytic degradation. Mild degradation of the drug substance was observed in base hydrolysis and considerable degradation observed during oxidative stress. The chromatographic method was fine tuned using the samples generated from forced degradation studies. Good resolution between the peaks corresponds to degradation products and the analyte was achieved on Agilent Zorbax SB-CN (250 mm x 4.6 mm, 5 microm) column. The mobile phase consists of a mixture of aqueous potassium di hydrogen ortho phosphate (pH 3.4), acetonitrile and methanol. The stress sample solutions were assayed against the qualified reference standard of rizatriptan benzoate and the mass balance in each case was close to 99.7% indicating that the developed method is stability indicating. Validation of the developed method was carried out as per ICH requirements.

Chromatography, High Pressure Liquid↗
PubMed1988

Gastrosphincter of Oddi reflex.

Gastric distention is known to stimulate gallbladder contraction as well as gastric acid and pancreatic exocrine secretion by way of neural reflexes. Gallbladder distention, in turn, has been shown to affect sphincter of Oddi motility. Since gastric distention may accompany endoscopic or operative biliary manometry, we tested the hypothesis that gastric distention alters sphincter of Oddi motility. In the prairie dog model, gastric distention with acid (0.1 M hydrochloric acid, pH 1.3) and alkaline (10(-5) sodium hydroxide, pH 8.8) isotonic saline solutions both resulted in significant increases in sphincter of Oddi phasic wave frequency, amplitude, and motility index. Similarly, gallbladder pressure increased during both distention periods, thus confirming the previously described pylorocholecystic reflex. These responses were abolished by systemic pretreatment with atropine, suggesting that this reflex is cholinergically mediated. These data suggest the presence of a gastrosphincter of Oddi reflex whereby gastric distention stimulates sphincter of Oddi motility in the prairie dog. We conclude that gastric distention is an important variable to be controlled when performing endoscopic or operative sphincter of Oddi manometry.

Ampulla of Vater↗
PubMed1977

Preparative countercurrent chromatography with a slowly rotating helical tube.

The capability of a simple countercurrent chromatographic scheme for obtaining high-resolution preparative-scale separations was demonstrated with the separations of a series of dinitrophenyl (DNP) amino acids and peptides. Basic studies on stationary phase retention and partition efficiency with a low-viscosity chloroform-acetic acid-0.1 N hydrochloric acid (2:2:1) phase system together with the results previously obtained with a vascous n-butanol system suggest a general applicability of low-interfacial tension phase systems by the present method.

Amino Acids↗
PubMed1989

Improved high-speed counter-current chromatograph with three multilayer coils connected in series. I. Design of the apparatus and performance of semipreparative columns in 2,4-dinitrophenyl amino acid separation.

A compact desktop model of a high-speed counter-current chromatograph holds three identical multilayer coils in the symmetrical positions around the rotary frame to maintain perfect balance of the centrifuge system without the use of a counterweight. These multilayer coils are connected in series to make up a total capacity of 400 ml while the unique gear arrangement on the rotary frame establishes a twist-free mechanism of the flow tubes so that continuous elution can be performed without the use of rotary seal. The high performance of the present system was successfully demonstrated in separations of 10-250 mg of 2,4-dinitrophenyl amino acid mixtures in a two-phase solvent system composed of chloroform-acetic acid-0.1 M hydrochloric acid (2:2:1, v/v/v).

Amino Acids↗
PubMed1986

New angle rotor coil planet centrifuge for counter-current chromatography. II. Design of the apparatus and studies on phase retention and partition capability.

A new angle rotor coil planet centrifuge (psi = 25 degrees), which produces a Type J-L synchronous planetary motion, has been constructed to examine its capability in terms of stationary phase retention and solute partitioning. Studies on phase distribution diagrams obtained from various two-phase solvent systems indicated that the present system can be adapted to a wide variety of solvent systems by adjusting the centrifugal conditions. Excellent partition capability of the apparatus was successfully demonstrated in separations of dinitrophenyl amino acid samples with chloroform-acetic acid-0.1 N hydrochloric acid (2:2:1).

Amino Acids↗