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Biomedical subjects

M T Kelly

Publications and source records attributed to M T Kelly.

At least 37 records · Page 2Linked to original sources

Comparison of two extraction methods for determination of propranolol and furosemide in human plasma by mixed-mode chromatography.

An isocratic high performance liquid chromatographic method is described for the determination of the beta-adrenergic blocking drug, propranolol, and the diuretic, furosemide, in human plasma. The two compounds and the internal standard were extracted from plasma using a two-step extraction technique. Propranolol and pindolol (internal standard) were first extracted from alkaline plasma into diethyl ether; this was followed by extraction of furosemide into acidified ether: hexane (65:35). The two extracts were then combined and evaporated under nitrogen, and the reconstituted residues were analysed on a C18/SCX reversed-phase/cation exchange column with a mobile phase of acetonitrile: 0.1 M sodium acetate pH 4 (33:67). The drugs and the internal standard were detected by UV absorption at 230 nm. The drugs were also extracted from plasma by a column-switching technique utilizing a ten-port valve. The drug compounds were retained on a C18 pre-column. A comparison of RSD for within-batch (intra-assay) and between-batch (inter-assay) runs for both methods was carried out, the liquid/liquid extraction method giving better recovery values. The calibration graphs were linear from 25-300 ng ml-1 for furosemide and 50-400 ng ml-1 for propranolol. Recovery values were > 90.0% by liquid/liquid extraction and > 76.0% by column switching.

Adrenergic beta-Antagonists↗

High-performance liquid chromatographic determination of taurine in human plasma using pre-column extraction and derivatization.

Plasma samples (100 microliters) were treated with 150 microliters of acetonitrile and centrifuged at 5800 g for 10 min and 50 microliters of 10 mM borate buffer (pH 9.2) were added to the supernatant solution. This was followed by the addition of a 50 microliters aliquot of 5 mM fluorescamine in acetonitrile and immediate vortex mixing. A 20 microliters sample was injected on to a reversed-phase HPLC system using a Bondclone C-18 10 microns analytical column (300 mm x 3.9 mm). The mobile phase was tetrahydrofuran-acetonitrile-phosphate buffer (15 mM, pH 3.5) (4:24:72, v/v/v). The taurine derivative was detected by measuring the UV absorbance of 385 nm. Platelet-poor plasma samples were spiked with known amounts of taurine and inter- and intra-assay calibration curves were obtained. The method was applied to the determination of taurine in platelet-rich plasma.

Chromatography, High Pressure Liquid↗

Determination of cimetidine in human plasma by high-performance liquid chromatography following liquid-liquid extraction.

A new method is described for the determination of cimetidine in human plasma. The drug and internal standard (ranitidine) were separated on a Nucleosil C18 5 microns (25 x 4.6 mm I.D.) column using a mobile phase of acetonitrile-phosphate buffer, pH 6.2 (25:75, v/v) containing 2.5 g/l heptane sulphonic acid. The mobile phase was delivered at a flow-rate of 0.9 ml/min, detection was by ultraviolet absorption at 228 nm and concentrations were calculated on the basis of peak areas. The drugs were extracted from alkaline plasma into ethyl acetate using a salting out procedure which involved the addition of 100 ml of a saturated solution of K2CO3 to each 250-microliters plasma aliquot. The method was validated over the concentration ranges 50-3000 ng/ml and 100-7000 ng/ml for two separate studies. Mean coefficients of variation were less than 6% for both intra- and inter-assay in both studies and recoveries varied between 71 and 81%. The method was successfully applied to the determination of cimetidine in plasma for a pharmacokinetic study.

Adolescent↗

Absorbed dose in the skin from beta emitters in medical and laboratory containers.

The International Commission on Radiation Protection have recently recommended an annual dose limit for the skin of radiation workers of 500 mSv at a depth of 20-100 microns averaged over any 1 cm2 regardless of the area exposed. It has previously been shown by the authors that beta dose rates on the outer surfaces of typical laboratory containers (vials, test tubes) or on medical syringes can exceed 100 mSv h-1 for radionuclide concentrations of the order of 1 MBq g-1, depending on container diameter, wall thickness and material and the beta particle energy spectrum. Since the fingers are frequently in contact with such containers it is of some importance to extend these dose calculations to depths below the skin surface, taking into consideration the anatomy of skin on the fingers. Using an extension of a Monte Carlo method previously described, dose rates have been calculated for the clinically useful radionuclides 90Y, 32P, 198Au, 153Sm and 131I. For polypropylene syringes the beta dose rates at a depth of 270 microns (a typical basal cell depth in the fingers) range from 77 to 135 mGy h-1 per MBq g-1 for 90Y (maximum energy 2.27 MeV) and approximately zero to 0.62 mGy h-1 per MBq g-1 for 131I (maximum energy 0.61 MeV). These results emphasize the importance of adequate finger protection when using high energy beta emitters and especially for clinicians who typically inject specific activities of the order of 100 MBq g-1 of 32P; in such cases annual permissible dose rates are exceeded in a matter of minutes. It is recommended that a minimum of 5 mm perspex finger protection be used for 90Y and 32P.

Beta Particles↗

Determination of ivermectin in bovine plasma by column-switching LC using on-line solid-phase extraction and trace enrichment.

Ivermectin is a potent avermectin anthelmintic, derived from the soil organism Streptomyces avermitilis. Because of its potency, it is administered in very low doses (200 micrograms kg-1), giving rise to plasma levels in the ppb region, and hence sensitive methods are required for its determination in biological samples. A method is described for the determination of ivermectin in bovine plasma, based on a commercially available C18 column employing a mobile phase composed of acetonitrile-water-isopropanol (40:40:35, v/v/v). Extraction from plasma is carried out on-line using coupled-column technology; switching between the extraction and analytical columns is achieved using a six-port column-switching valve. The method is linear over the range 2-100 ng ml-1 (r = 0.9994) and has a limit of detection of 0.8 ng ml-1 (signal-to-noise ratio = 3). Reproducibility of the method, expressed as the overall mean relative standard deviation, is 6.7% (intra-assay) and 3.8% (inter-assay). The mean recovery of the drug from plasma is 76.4%. The method is simple and economical in its execution and is sufficiently reproducible to obviate the need for an internal standard.

1-Propanol↗

Analysis of terbutaline in human plasma by high-performance liquid chromatography with electrochemical detection using a micro-electrochemical flow cell.

A high-performance liquid chromatographic method is described for the determination of terbutaline in human plasma in the range 1-35 ng/ml. Detection was achieved using a carbon fibre micro-electrochemical detector and a column-switching system. The microelectrode cell has advantages over conventional glassy carbon electrode-based detection systems in that it is easy to prepare, flexible in its operation and suffers less trouble from problems such as air bubbles and leaks. Furthermore, it has a better detection limit for terbutaline (0.8 ng/ml) to that obtained using a conventional glassy carbon electrode flow detector (2 ng/ml). Sample clean-up was by on-line solid-phase extraction with column switching, providing a method which was sensitive and reproducible, where the mean overall coefficient of variation was 5.60% and drug recovery in excess of 86% at the concentration levels studied.

Calibration↗

Sodium hypochlorite disinfection of irreversible hydrocolloid impression material.

Alginate impression material is one of the most frequently used in dentistry. However, this material is susceptible to dimensional distortion during disinfection because of its hydrophilic nature. This study examined the effects of alginate disinfection using a sodium hypochlorite spray or impression immersion. Spray disinfection of an alginate impression did not cause dimensional differences of the poured stone casts when compared with casts from water-rinsed controls. Immersion disinfection created dimensional distortion of the anterior, posterior, and interarch model segments. Both the spray and immersion treatments equally decreased the surface detail reproducibility. The antimicrobial effects of the spray treatment were similar to those of the immersion treatment, while mere water rinsing resulted in inadequate disinfection.

Aerosols↗

Thermal properties of a methyl methacrylate-based orthodontic bonding adhesive.

Methyl methacrylate-based (MMA-based) bonding resins have been used in orthodontics because they offer easy removal of both the bonded bracket and the residual adhesive at case completion. However, these materials are not cross-linked, and the brackets bonded with this type of product may undergo drifting when subjected to temperatures slightly higher than those in the mouth. This research investigated the influence of heat on the debonding characteristics of a MMA-based bonding resin compared with those of a BIS-GMA-based system. The temperature of initial bracket movement, as well as of final bracket displacement, was noted for a variety of applied loads (141, 226, 425, 934, and 1727 gm) with stainless steel brackets bonded to etched bovine enamel. The results showed that the MMA-based material underwent a glass transition near 47 degrees C in which the initiation of bracket drift resulted on the tooth surface. This temperature proved independent of the applied load. Further heating resulted in the release of heat from the resin as a result of further curing. The extent of bracket drift associated with this secondary heat release was dependent on the applied load. The debonding temperature of the BIS-GMA-based system was three to six times greater than that of the MMA-based product. Practitioners should be aware that brackets bonded with MMA-based resins have the potential for drifting when subjected to temperatures within the normal range of hot fluids consumed by their patients. Bracket drift could result in deactivation of orthodontic force and could prolong the treatment time.

Adhesives↗

Comparison of a direct fecal Shiga-like toxin assay and sorbitol-MacConkey agar culture for laboratory diagnosis of enterohemorrhagic Escherichia coli infection.

A direct fecal Shiga-like toxin assay (DSLTA) was used to prospectively screen 9,449 unselected stool samples, received at the British Columbia Provincial Health Laboratories and the Metropolitan Laboratories of Vancouver, for Shiga-like toxin I and Shiga-like toxin II. The results were compared with results of routine stool culture on sorbitol-MacConkey agar (SMAC) for Escherichia coli O157:H7. Of 80 specimens positive by either method, 59 (74%) and 74 (93%) were positive by SMAC and DSLTA, respectively; 53 (66%) were positive by both methods, 21 (26%) were positive by DSLTA only, and 6 (7%) were positive by SMAC only. On further screening, Shiga-like toxin-producing E. coli were detected in 8 (38%) of the 21 stools positive by DSLTA only, including serotypes O157:H7 (1 stool), O26:K60 (5 stools), O128:K67 (1 stool), and O103:H2 (1 stool). For the remaining 13 stools in which no SLTEC was found but DSLTA was positive, clinical information revealed that 11 of 12 patients had diarrheal illnesses, and 4 of these 11 had bloody diarrhea or hemolytic-uremic syndrome. Stools positive only by SMAC were collected earlier in the illness than stools positive by DSLTA, suggesting that free fecal toxin levels may be too low to detect at this time. Overall we found that DSLTA detected 19% more positive specimens than SMAC and that Shiga-like toxin-producing E. coli serotypes other than E. coli O157:H7 are causing disease in the province of British Columbia, Canada.

Bacterial Toxins↗

Evaluation of the Autoscan Walkaway system for rapid identification and susceptibility testing of gram-negative bacilli.

We evaluated the performance of the Autoscan Walkaway (W/A) system (MicroScan, Sacramento, Calif.) in conjunction with the fluorometric Neg Combo panels for rapid identification and susceptibility testing of gram-negative bacilli. Fermentative and nonfermentative gram-negative bacilli were tested in parallel with the W/A system and the Cathra Repliscan replicator (C/R) system (Cathra, St. Paul, Minn.). Conventional biochemical testing and agar dilution testing were used to resolve the identification and susceptibility testing discrepancies. Of 495 clinical isolates tested, 445 (90%) were correctly identified by the W/A system and 483 (98%) were correctly identified by the C/R system. Repeat testing by using updated versions of the W/A system's computer identification software failed to demonstrate improved identification accuracy. For susceptibility testing, the W/A system demonstrated 5.6% total interpretative category errors, including only 0.9% major and very major errors. The comparative C/R system produced only 1% errors overall, including 0.2% major and very major errors. Although the W/A system is highly automated and is capable of producing results rapidly, our findings suggest that additional identification and susceptibility testing refinements are needed before the system will be suitable for routine use.

Anti-Bacterial Agents↗

Characterization of a DNA probe for detection of Mycobacterium tuberculosis complex in clinical samples by polymerase chain reaction.

We cloned and sequenced a DNA fragment from Mycobacterium tuberculosis for use in the identification of members of the M. tuberculosis complex. The DNA probe for culture confirmation had a sensitivity and a specificity of 100%. By using primers developed from this probe, the polymerase chain reaction detected 20 mycobacteria by ethidium bromide staining. This polymerase chain reaction system demonstrated 98% sensitivity and 100% specificity for detection of the M. tuberculosis complex in 200 sputum specimens.

Bacteriological Techniques↗

Biochemical characteristics and plasmids of clinical and environmental Plesiomonas shigelloides.

The biochemical characteristics of 68 clinical and 5 environmental Plesiomonas shigelloides isolates were determined. The reactions for ONPG, phenylalanine deaminase, motility, lactose fermentation and salicin fermentation were different from previously published findings. The reactions of the clinical and environmental isolates were very similar. The isolates were also tested for the presence of plasmids, and 40% of the strains were found to harbor between one and seven plasmids. The findings of this study will be useful for the identification and characterization of this recently recognized enteric pathogen.

Animals↗

High-performance liquid chromatographic determination of nifedipine, nicardipine and pindolol using a carbon fibre flow-through amperometric detector.

The electrochemical properties of the calcium-channel blockers, nifedipine and nicardipine, and the beta-blocking agent, pindolol, have been exploited for the determination of their concentrations in plasma samples. High-performance liquid chromatography (HPLC) separation was carried out on a cyanopropyl modified column and the drugs were detected in a flow-through carbon fibre microelectrode cell. The chromatographic system was coupled to a column-switching arrangement in order to perform on-line solid-phase extraction of the drugs from spiked human plasma. Preliminary investigations showed the response of the method to be linear over a range of 20-500 ng ml-1 in plasma with a limit of detection of approximately 15 ng ml-1 for each compound.

Carbon↗

Etiology of childhood diarrhea in Beijing, China.

To determine the role of recently recognized enteropathogens in childhood diarrhea in China, 221 children with diarrhea and 108 controls seen at the Beijing Children's Hospital were studied during April and May 1989. Stools were examined for ova, parasites, and rotavirus, cultured for bacterial pathogens, and probed for enterotoxigenic Escherichia coli (ETEC), enteroinvasive E. coli (EIEC), enterohemorrhagic E. coli (EHEC), and enteropathogenic adherence factor-positive (EAF+) E. coli. Pathogens were identified in 56.5% of children with diarrhea and 43.5% of controls (P = 0.04). Detection of enteropathogens was significantly greater in patients examined within 1 week of symptom onset (65%) than in patients examined later (39%; P = 0.01). ETEC was the most frequently detected pathogen in children with diarrhea, accounting for 20% of the cases. Other agents identified in patients included the following: salmonellae, 12%; rotavirus, 7%; EIEC, 7%; EHEC, 7%; members of the Aeromonas hydrophila group, 6%; EAF+ E. coli, 5%; Ascaris lumbricoides, 3%; shigellae, 3%; campylobacters, 2%; and Vibrio spp., 0.5%. The isolation rates of salmonellae (P = 0.02), EAF+ E. coli (P = 0.04), and mixed pathogens (P = 0.05) were significantly greater for diarrhea patients than for controls. Resistance to multiple antimicrobial agents occurred in 39% of the Salmonella isolates, 22% of the Aeromonas isolates, and 17% of the Shigella isolates. Multiresistant salmonellae (P = 0.05) and shigellae were recovered from diarrheal stools only. Ciprofloxacin, cefotaxime, and imipenem were the only agents tested to which all bacterial isolates were susceptible in vitro. These results suggest that both traditional and newly recognized agents are important causes of childhood diarrhea in Beijing and that therapy may be complicated by indigenous antimicrobial resistance.

Child, Preschool↗

High-performance liquid chromatographic method for the comparison of tanning capacity of tannic acid batches used in the manufacture of pregnancy testing kits.

A high-performance liquid chromatographic (HPLC) method was developed for the quantitative comparison of various batches of tannic acid from the same manufacturer used to aid the binding of human chorionic gonadotropin to sheep erythrocytes in the manufacture of pregnancy testing kits. The tannic acids were separated by reversed-phase HPLC on a C18 column using gradient elution with aqueous methanolic eluents at low pH. A portion of the chromatogram corresponding to the compounds involved in the tanninisation process was integrated and a linear relationship was established between this peak area and tannic acid concentration. The correlation coefficient was greater than 0.993 even in the absence of an internal standard. Tanning capacity was evaluated on the basis of the amount of tannic acid which remained following incubation with a known quantity of erythrocytes. The application of this procedure to three batches of tannic acid is demonstrated.

Chromatography, High Pressure Liquid↗

Evaluation of drug-free plasma profiles by high-performance liquid chromatography following on-line solid-phase extraction.

The effect of varying the type of column and eluent composition on drug-free plasma profiles was investigated. The study was based on a C18 and a CN column; methanol and acetonitrile were the organic modifiers used. The plasma profiles were evaluated quantitatively by measuring the number of interfering peaks greater than 8 . 10(-4) absorbance units in the area of interest along the chromatogram. Results were subjected to statistical treatment using a three-factor analysis of variance design. The three factors were the column, the type of organic modifier and either the percentage organic modifier, the pH or the ionic strength. Analysis of the data revealed that significant effects were seen with changing eluent composition, particularly with regard to the percentage of organic modifier, and that the observed effects were strongly dependent on the type of column and the type of organic modifier under consideration.

Acetonitriles↗