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PubMed1998

Toroidal coil counter-current chromatography. Achievement of high resolution by optimizing flow-rate, rotation speed, sample volume and tube length.

This paper deals with optimization of a new seal-free compact toroidal coil centrifuge to achieve high resolution in analytical counter-current chromatography (CCC). Toroidal coil CCC (hydrostatic motion) has advantages compared with high-speed CCC (efficiently mixing solution with planetary motion) in the separation of protein or easily emulsified samples. A toroidal coil separation column of 0.4 mm I.D. PTFE tubing was accommodated around the periphery of the cylindrical centrifuge bowl. Using a two-phase solvent system composed of chloroform-acetic acid-0.1 M hydrochloric acid (2:2:1, v/v/v) and a set of dinitrophenyl-amino acids as test samples, a series of experiments was performed with parameters such as the column length, sample volume, flow-rate, elution mode of the mobile phase and rotation speed. The highest efficiency, over 10,000 theoretical plates, was achieved with a 100 m long coiled tube and an 11 ml total capacity at a flow-rate of 0.01 ml/min at 800 rpm.

Amino Acids↗
PubMed1982

Specimen handling and high-performance liquid chromatographic determination of furosemide.

A simple high-performance liquid chromatographic method to measure furosemide in plasma and urine is described. Furosemide fluoresces best, but is unstable, at acidic pH and is subject to photochemical degradation. These factors were analysed and the results prompted changes in previously described methods. All specimens were very carefully protected from light; extraction and acidification were done with acetic acid instead of hydrochloric acid. With these precautions no 4-chloro-5-sulphamoylanthranilic acid was found in biological specimens. The main metabolite was furosemide glucuronide (20% of furosemide excretion). Sensitivity was 0.1 and 0.5 microgram/ml for plasma and urine, respectively. The applicability of our method for furosemide studies is demonstrated.

Adult↗
PubMed2002

Pharmaceutical development of a parenteral lyophilized formulation of the antimetastatic ruthenium complex NAMI-A.

This paper describes the development of a stable pharmaceutical dosage form for NAMI-A, a novel antimetastatic ruthenium complex, for Phase I testing. NAMI-A drug substance was characterized using several spectrometric and chromatographic techniques. In preformulation studies, it was found that NAMI-A in aqueous solution was not stable enough to allow sterilization by moist heat. The effect of several excipients on the stability of the formulation solution was investigated. None of them provided sufficient stability to allow long-term storage of an aqueous solution of NAMI-A. Therefore, a lyophilized product was developed. Five different formulations were prepared and subjected to thermogravimetric (TG) analysis and stability studies at various conditions for 1 year. Minimal degradation during the production process is achieved with a formulation solution of pH 3-4. Of the acids tested, only hydrochloric acid (HCl 0.1 mM) both stabilized the formulation solution and was compatible with the lyophilized product. This product was stable for at least 1 year when stored at -20 degrees C, 25 degrees C/60% relative humidity (RH) and 40 degrees C/75% RH, and was also photostable.

Antineoplastic Agents↗
PubMed2001

Oxidative degradation study of nitrendipine using stability indicating, HPLC, HPTLC and spectrophotometric method.

The objective of this study is to develop validated stability indicating HPLC (A), HPTLC (B) and spectrophotometric (C) method for the estimation of nitrendipine. The stability indicating capability of the assays is proved using forced degradation, by exposing drug solution to sunlight, acidic and alkaline medium. The chromatogram and UV spectrum showed nitrendipine well resolved from the degradation product. Degradation of drug is found faster in acidic (0.1 N hydrochloric acid) medium as compared to alkaline (0.1 N sodium hydroxide) medium at 100 degrees C. Also, photodegradation is studied, with special emphasis on the effect of solvents like methanol (1), chloroform (2), dichloromethane (3), acetone (4) and ethyl acetate (5), on the rate of photodegradation. The degradation of title compound followed first order kinetics in all cases. Estimation of the drug is carried out by the stability indicating methods mentioned, using one point standardization within the linearity range of interest. The methods are compared in respect of performance precision and accuracy. Major route of degradation in all cases is found to be oxidation and degradation product is confirmed as dehydronitrendipine by the use of relevant UV, IR and 1H NMR spectrometry.

Chromatography, High Pressure Liquid↗
PubMed2000

Electrospray ionization tandem mass spectrometry for structural elucidation of protonated brevetoxins in red tide algae.

Brevetoxins, the toxic components of "red tide" algae, all share one of two robust polycyclic ether backbone structures, but they are distinguished by differing side-chain substituents. Electrospray ionization mass spectrometry analyses of brevetoxins have shown that the polyether structure invariably has a very high affinity for sodium cations that results in the production of abundant (M + Na)+ ions even when sodium cations are only present as impurities. Because the ionic charge tends to remain localized on the sodium atom and because at least two bonds must be broken in order to produce polycyclic backbone fragmentation, it is extremely difficult to obtain abundant product ions (other than Na+) from (M + Na)+ brevetoxin precursor ions in low-energy collision-induced dissociation (CID) MS/MS experiments. This report establishes that acid additives (oxalic acid, trifluoroacetic acid, and particularly hydrochloric acid) in aqueous methanol solutions can promote high yields of protonated brevetoxin molecules (MH+ ions) for Btx-1, -2, and -9 brevetoxins. Most importantly, unlike their (M + Na)+ counterparts, MH+ precursor ions offer readily detectable product ions in CID MS/MS experiments, even under low-energy collisions. This direct structural characterization approach has provided decomposition information from brevetoxins that was previously inaccessible, including the identification of diagnostic product ions for "type A" brevetoxins (m/z 611) and "type B" brevetoxins (m/z 779, 473, 179) and characteristic ions for Btx-1 (m/z 221, 139), Btx-2 (m/z 153), and Btx-9 (m/z 157, 85). Precursor ion scans and constant neutral loss scans are proposed to enable screening of individual type A or type B brevetoxins present in naturally occurring mixtures.

Animals↗
PubMed2002

Carbon nanotube-modified electrodes for the simultaneous determination of dopamine and ascorbic acid.

The voltammetric separation of dopamine and ascorbic acid was studied with cyclic voltammetry at two kinds of carbon nanotube-modified electrodes (coated and intercalated). The anodic peak difference reached 270 mV under the present conditions. The separation mechanism and effect factors were carefully studied. Using various types of surfactants as coating dispersants of carbon nanotubes, it was demonstrated that the charge nature of the surfactants had a strong effect on the electrochemical behavior of dopamine and ascorbic acid. When the oxidation solution of carbon nanotubes was changed from the most commonly used mixed concentrated nitric acid and sulfuric acid (1 + 3 v/v) to dilute nitric acid and to hydrochloric acid, the anodic peak separation value of dopamine and ascorbic acid increased significantly, and it was shown that carboxylic acid groups attached to the carbon nanotubes were an adverse factor for the discrimination of DA from AA. These results indicated that the resolution of DA and AA was mainly attributable to the stereo porous interfacial layer formed from aggregated pores and inner cavities of the carbon nanotubes. The modified electrodes exhibited an attractive ability to measure DA and AA simultaneously and showed good stability and reproducibility.

Ascorbic Acid↗
PubMed2002

Influence of intestinal anaerobes and organic acids on the growth of enterohaemorrhagic Escherichia coli O157:H7.

A suspension of human faeces (FS) and its anaerobic culture (FC), bacterial metabolic products and organic acids were examined for inhibitory effects on growth and verotoxin 2 (VT2) production of Escherichia coli O157:H7 in vitro. FS and FC showed a marked inhibitory activity to growth and production of VT2 by E. coli O157:H7 under anaerobic conditions. They may have both bacteriostatic and bactericidal effects on E. coli O157. The growth of E. coli O157 was markedly suppressed by acetic, propionic and butyric acids compared with hydrochloric acid and lactic acid at concentrations between 25 mM and 40 mM, being proportional to the pH values. At pH 5.5, 40 mM of short-chain fatty acids (SCFAs) almost completely inhibited the growth of E. coli O157. SCFAs markedly inhibited the growth of E. coli O157 at pH 6.0 rather than pH 7.0. Propionic acid is likely to be more suppressive to E. coli than acetic and butyric acids. The production of VT2 was approximately proportional to the growth of E. coli O157. However, incubation for 24 h in vitro showed that the growth and VT2 production of E. coli O157 decreased but were not completely inhibited at pH 6.5 and 7.0 in a mixture of acetic, propionic and butyric acids at a physiological concentration (110 mM, 60:25:25, respectively, in molar ratio). It is probable that the colonic microflora could contribute to a reduction of E. coli O157:H7 infections via the activation of intestinal fermentation by dietary manipulation or something similar to give pH 6.0 or <6.0 and that factors such as age, chemical therapy and body condition, which have effects on the balance of the intestinal microflora, would be associated with the incidence rates of E. coli O157 infections.

Acetic Acid↗
PubMed1976

Comparison of different purification procedure for extraction of staphylococcal enterotoxin A from foods.

Different procedures commonly used for extraction, purification, and concentration of staphylococcal enterotoxins from foods were investigated with 131I- and 125I-labeled staphylococcal enterotoxin A. Loss of labeled enterotoxin A was compared with loss of total nitrogen. The results showed that in most of the common procedures, such as gel filtration, ion exchange, and heat treatment, the percentage of loss of labeled enterotoxin A was greater than the loss of total nitrogen. Chloroform extraction and acid precipitation with hydrochloric acid had nearly the same effect on the purification of both labeled enterotoxin A and total nitrogen. Ammonium sulfate precipitation proved to be practical and was successfully used for purification of enterotoxin A from sausage extract. Simultaneous use of trypsin and Pseudomonas peptidase for treatment of food extracts considerably reduced food proteins capable of interfering with serological detection of enterotoxins but did not essentailly influence the loss of enterotoxin A.

Enterotoxins↗
PubMed1986

Is urea formation regulated primarily by acid-base balance in vivo?

Large quantities of ammonium and bicarbonate are produced each day from the metabolism of dietary protein. It has recently been proposed that urea synthesis is regulated by the need to remove this large load of bicarbonate. The purpose of these experiments was to test whether the primary function of ureagenesis in vivo is to remove ammonium or bicarbonate. The first series of rats were given a constant acid load as hydrochloric acid or ammonium chloride; individual rats received a constant nitrogen load at a time when their plasma acid-base status ranged from normal (pH 7.4, 28 mM HCO3) to severe metabolic acidosis (pH 6.9, 6 mM HCO3). Urea plus ammonium excretions and the blood urea, glutamine, and ammonium concentrations were monitored with time. Within the constraints of non-steady-state conditions, the rate of urea synthesis was constant and the plasma glutamine and ammonium concentrations also remained constant; thus it appears that the rate of urea synthesis was not primarily regulated by the acid-base status of the animal in vivo over a wide range of plasma ammonium concentrations. In quantitative terms, the vast bulk of the ammonium load was converted to urea over 80 min; only a small quantity of ammonium appeared as circulating glutamine or urinary ammonium. Urea synthesis was proportional to the nitrogen load. A second series of rats received sodium bicarbonate; urea synthesis was not augmented by a bicarbonate load. We conclude from these studies that the need to dispose of excess bicarbonate does not primarily determine the rate of ureagenesis in vivo. The data support the classical view that ureagenesis is controlled by the quantity of ammonium to be removed.

Acid-Base Equilibrium↗
PubMed2005

Safety pharmacology of sibutramine mesylate, an anti-obesity drug.

Sibutramine mesylate is a new anti-obesity drug. It is a crystalline salt of sibutramine developed to improve the solubility of sibutramine hydrochloride. Methanesulfonic acid was used as a salt-forming acid instead of hydrochloric acid, resulting in a greatly improved solubility of 1000 mg/mL in water. Sibutramine mesylate was administered orally to ICR mice, Sprague-Dawley rats, and beagle dogs at dose levels of 1.15, 3.45, and 11.50 mg/kg to measure its effects on the central nervous system (CNS), general behaviour, cardiovascular-respiratory system and the other organ systems. Following administration of sibutramine mesylate, spontaneous locomotor activity was significantly increased from 120 min to 24 hours at 3.45 mg/kg and from 30 min to 24 hours at 11.50 mg/kg. Furthermore, there were a decrease in hexobarbital-induced sleep time, an increase in respiratory rate at 120 min, increases in intestinal transport capacity and gastric pH at 11.50 mg/kg, and decreases in gastric volume and total acidity at 3.45 and 11.50 mg/kg. However sibutramine mesylate caused no effects on general behaviour, motor coordination, body temperature, analgesia, convulsion, blood pressure, heart rate, electrocardiogram, cardiac functions of the isolated rat heart, isolated smooth muscles and renal function. Based on the above results, it was concluded that sibutramine mesylate caused effects on the spontaneous locomotor activity, hexobarbital-induced sleep time, respiration, gastrointestinal transport, and gastric secretion at a dose level of 3.45 mg/kg or greater but caused no effects on other general pharmacological reactions.

Administration, Oral↗
PubMed2002

Synthesis of optically active homocysteine from methionine and its use in preparing four stereoisomers of cystathionine.

In order to synthesize four stereoisomers of cystathionine (CYT), D- and L-homocysteines (D- and L-Hcy) were synthesized from methionine (Met) by a facile procedure. L-Met was reacted with dichloroacetic acid in concentrated hydrochloric acid under reflux to give (4S)-1,3-thiazane-2,4-dicarboxylic acid hydrochloride [(4S)-TDC.HCl]. L-Hcy was obtained by treatment of (4S)-TDC.HCl with hydroxylamine. D-Hcy was also synthesized from D-Met via (4R)-TDC.HCl intermediate. The obtained D- and L-Hcy were condensed with (R)- and (S)-2-amino-3-chloropropanoic acid hydrochlorides under alkaline conditions to give four stereoisomers of CYT.

Cystathionine↗
PubMedDate not supplied

Retention of chromium (VI) on a macroporous char following ChemChar gasification and successive leaching with water and acids.

A granular macroporous char, triple-reverse-burn (TRB) char, was loaded with 23.40 mg Cr/g TRB char from an aqueous solution, and the retained metal was leached by water, 0.66 M nitric acid, concentrated nitric acid, and concentrated hydrochloric acid before and after treatment by a reductive thermal gasification process (ChemChar process developed by ChemChar Research, Inc., Columbia, Missouri). The chromium leachate was analyzed by flame atomic adsorption. Reverse- and forward-mode gasifications were performed on the metal-laden char. With the exception of a 10% mass loss of carbon, the reverse mode gasification process does not change the physical characteristics of the granular char, but does increase the retention of the chromium from 16.7 to 24.2%, depending on the leachant. The forward mode gasification process produces a vitrified (or glasslike) ash residue. There was an 11.6 to 13.1% increase in the retention of the chromium by the slag and ash when compared to the nongasified chromium-loaded TRB char. Chromium (VI) was effectively removed from solution by TRB char and found to be retained to a higher degree on the char after a reductive thermal treatment.

Adsorption↗
PubMed2004

Effect of different pharmacologic and chemical agents on the integrity of hydatid cyst membranes.

OBJECTIVE: We performed an in vitro investigation of the effects of widely used scolicidal and sclerosing agents, as well as some pharmacologic products, on the integrity of the membrane of hydatid cysts. MATERIALS AND METHODS: Two milliliters each of 22 agents, 2 mL of clear fluid, and one piece of hydatid cyst membrane were put into bottles. The hydatid cyst membranes were evaluated by visual observation and manual palpation. Visual examination of the bottles was performed daily for 7 days, and observations of membrane changes, including translucency, destruction, swelling, and melting, were recorded. Manual evaluation was done on the seventh day by finger examination, and membrane fragility was scored. RESULTS: The hydatid cyst membrane was completely melted in a few minutes in a 2.5% solution of sodium hypochlorite and in 1 hr by a 0.1% sodium hypochlorite solution. The integrity of the hydatid cyst membrane was preserved in alcohol, acetone, glutaraldehyde, albendazole, acetylsalicylic acid, formaldehyde, lidocaine, hydrochloric acid, ammonia, pancreatin, Betadine, methylene blue, and isotonic saline samples. The membranes in the metronidazole and hypertonic saline solutions were not damaged but showed significantly increased fragility. The membranes in levamisole and piperazine hexahydrate became translucent and showed moderate fragility. CONCLUSION: None of the agents that are used in clinical practice had important effects on the dissolution of hydatid cyst membranes. However, sodium hypochlorite solutions completely melted the hydatid cyst membranes. Because the use of this agent on living tissue is limited, further study is needed to investigate its clinical use.

Animals↗
PubMed1977

Effect of ultra-high-temperature steam injection processing on sulfur-containing amino acids in milk.

Raw skim milk was processed by a modified No-Bac Unitherm IV System (Cherry-Burrell Corp.) at 143 C for 8 s, vacuum cooled to 71 C, collected, and cooled to 4 C. Raw and ultra-high-temperature processed skim milks were oxidized with performic acid, hydrolyzed with hydrochloric acid, and analyzed for cysteine and cystine (as cysteic acid) and methionine (as methionine sulfone) on a Beckman Automatic Amino Acid Analyzer. A loss of approximately 34% of these amino acids was observed in ultra-high-temperature processed skim milk. Sulfhydryl and disulfide groups determined with Ellman's reagent [5,5'-dithiobis(2-nitrobenzoic acid)] indicated no free sulfhydryls in raw skim milk and .07 mmole per liter in ultra-high-temperature processed skim milk. Loss in total sulfhydryl and disulfide groups was approximately 16% in ultra-high-temperature processed milk. Volatile sulfur compounds were detected by odor in the vapor collected from the vacuum chamber, but they could not be identified. Amino acid analysis of milk deposit collected from the injection section revealed .09 microgram of cysteine and cystine (as cysteic acid) per gram of deposit and no detectable methionine.

Amino Acids, Sulfur↗
PubMed2005

Design and development of low cost, simple, rapid and safe, modified field kits for the visual detection and determination of arsenic in drinking water samples.

Arsenic is naturally found in surface and ground waters and the inorganic forms of arsenic are the most toxic forms. The adverse health effects of arsenic may involve the respiratory, gastrointestinal, cardiovascular, nervous, and haematopoietic systems. Arsenic contamination in drinking water is a global problem widely seen in Bangladesh and West Bengal of the Indian sub continent. As there is a great demand for field test kits due to the anticipated reduction of the US EPA arsenic standard from 50ppb to 10ppb a field kit which offers rapid, simple and safe method for precise estimation of arsenic at 10ppb in drinking water samples is developed. Field methods, based on the mercuric-bromide-stain, consist of three different major parts, which are carried out stepwise. The first part of the procedure is to remove serious interference caused by hydrogen sulphide. In commercially available kits either the sulphide is oxidized to sulphate and the excess oxidizing reagent removed prior to the hydride generation step or, the hydrogen sulphide is filtered out by passing the gas stream through a filter impregnated with lead acetate during the hydride generation step. The present method employs cupric chloride in combination with ferric chloride or Fentonis reagent for the removal of hydrogen sulphide, which is rapid, simple and more efficient. Other interferences at this step of the analyses are normally not expected for drinking water analysis. In the second step, the generation of the arsine gas involves the classical way of using zinc metal and hydrochloric acid, which produce the enascenti hydrogen, which is the actual reducing agent. Hydrochloric acid can be replaced by sulfamic acid, which is solid and avoids a major disadvantage of having to handle a corrosive liquid in the field. The arsine gas produces a yellowish spot on the reagent paper. Depending on the arsenic content, either, Yellow n H (HgBr)2 As (10-50ppb), Brown n (HgBr)3 As (50-100ppb) or Black n Hg3 As2 (>100ppb) are formed which can be precisely estimated by visual comparison with standard color chart. The results obtained by field kits agree well with the data obtained through I.C.P.AES methods. The most important characteristic for field measurement is that analytical results can be obtained on the site where the sample is taken with high precision and can be conveniently utilized for monitoring arsenic rapidly in a highly contaminated large geographical area.

Arsenic↗
PubMed1992

[Diagnosis, countermeasure and classification of acidosis].

Acidosis is the result of the net addition of hydrogen ion to the extracellular space or loss of bicarbonate from that space. Hydrogen ion may be added by the increased production of strong acids, an increase in CO2 concentration, or the addition of exogenous acids. The common approach to the differential diagnosis of metabolic acidosis is to divide the patients into two categories based on whether the anion gap in plasma is increased or not. Metabolic acidosis with normal anion gap (hyperchloremic) suggests that bicarbonate has been effectively replaced by chloride. In contrast, metabolic acidosis with an increased anion gap suggests addition to the body fluids of an acid other than hydrochloric acids or its equivalent.

Acidosis↗
PubMed1992

Evaluation of a photodiode array/HPLC-based system for the detection and quantitation of basic drugs in postmortem blood.

A systematic analytical approach has been developed for liquid chromatography determination of a number of basic drugs in postmortem blood. Using selective extraction, that is, back extraction into 0.2N sulfuric acid and 6N hydrochloric acid after the initial extraction with toluene under basic conditions (from 2 mL of blood), basic and weakly basic drugs, such as propranolol and diazepam, can be simultaneously quantitated and identified with a high degree of confidence. A microcomputer-based photodiode array detector was used to evaluate peak purity and facilitate peak identification. An automatic library search was performed at the end of each analysis using the system software. The method was validated for within-day and between-day precision for ten basic drugs at two concentrations. The coefficient of variation for the between-day precision was less than 8.7%. Accuracy of the assay was tested at four concentrations using linear regression analysis. The coefficients of determination (r2) for all ten drugs were greater than 0.99, and their slopes were close to unity. The chromatographic conditions developed are suitable for the screening of several basic, acidic, amphoteric, and neutral drugs. Retention data and ultraviolet spectral data for 119 drugs on two reversed-phase columns, using acidic mobile phases, are also presented.

Blood Chemical Analysis↗
PubMed1991

Chemical forms of selenium in corn and rice grown in a high selenium area of China.

The chemical forms of selenium were determined in maize grown near Yutangba Village and in rice grown near Haubei village, Exi Prefecture, Enshi county of Hubei Province, China. The maize sample contained 18 ppm and the rice samples an average of 3.6 ppm selenium. After they were ground to obtain a fine flour, this was acid hydrolyzed with hydrochloric acid in an inert atmosphere and the hydrolyzates chromatographed on columns of Dionex DC6A resin. The results indicate that the majority of the selenium is present as selenomethionine in both rice and corn.

China↗