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Biomedical subjects

H G Fouda

Publications and source records attributed to H G Fouda.

30 records · Page 2Linked to original sources

Robotics in biomedical chromatography and electrophoresis.

The ideal laboratory robot can be viewed as "an indefatigable assistant capable of working continuously for 24 h a day with constant efficiency". The development of a system approaching that promise requires considerable skill and time commitment, a thorough understanding of the capabilities and limitations of the robot and its specialized modules and an intimate knowledge of the functions to be automated. The robot need not emulate every manual step. Effective substitutes for difficult steps must be devised. The future of laboratory robots depends not only on technological advances in other fields, but also on the skill and creativity of chromatographers and other scientists. The robot has been applied to automate numerous biomedical chromatography and electrophoresis methods. The quality of its data can approach, and in some cases exceed, that of manual methods. Maintaining high data quality during continuous operation requires frequent maintenance and validation. Well designed robotic systems can yield substantial increase in the laboratory productivity without a corresponding increase in manpower. They can free skilled personnel from mundane tasks and can enhance the safety of the laboratory environment. The integration of robotics, chromatography systems and laboratory information management systems permits full automation and affords opportunities for unattended method development and for future incorporation of artificial intelligence techniques and the evolution of expert systems. Finally, humanoid attributes aside, robotic utilization in the laboratory should not be an end in itself. The robot is a useful tool that should be utilized only when it is prudent and cost-effective to do so.

Chemistry, Clinical↗

Liquid chromatographic analysis of doxazosin in human serum with manual and robotic sample preparation.

A specific method for the determination of the antihypertensive drug doxazosin in human serum is described. The method utilizes the related drug prazosin as an internal standard and is based on a simple extraction scheme followed by analysis by reversed-phase ion suppression high-performance liquid chromatography (HPLC) on an alumina-based column with fluorescence detection. The method is completely automated with a flexible robotic system for the analysis of drugs in biological fluids. The robotic automation of the method allows a 20% increase in the sample throughput and the savings of about 7 man-hours a day. Both the manual and robotic procedures yield precise quantitative results over the therapeutically relevant concentration range of 0.5 to 20 ng/mL of serum.

Chemical Phenomena↗

Selected ion monitoring analysis of CP-55,940, a cannabinoid derived analgetic agent.

A method is described for the quantification in plasma of CP-55,940, a new non-opiate oral analgetic agent, using capillary gas chromatography/mass spectrometry. After extraction, derivation of the drug and a structural analog internal standard is carried out with trifluoracetyl imidazole. The derivative exhibited excellent gas chromatographic properties. Electron impact yielded an abundant high mass ion whose monitoring permitted the sensitive and specific determination of CP-55,940 down to 0.5 ng ml-1.

Analgesics↗

A gas chromatographic/mass spectrometric assay for flutroline, a gamma-carboline antipsychotic agent, with direct derivatization on a moving needle injector.

A method has been developed for the determination of flutroline in plasma using capillary gas chromatography and selected ion monitoring detection of the TMS derivative. The method is linear over the concentration range of 3-60 ng ml-1 and was used to define the drug pharmacokinetics and bioavailability in animals and man. A novel method for direct derivatization on the tip of a moving needle injector is described.

Antipsychotic Agents↗

Sensitive assay for determination of hydroxyzine in plasma and its human pharmacokinetics.

An assay suitable for hydroxyzine determination in human plasma following therapeutic doses was developed. The method involves GLC and chemical-ionization mass spectrometry of the acetate derivatives of hydroxyzine and of a pentadeuterated analog internal standard. Following administration of 100-mg single oral doses to normal male volunteers, peak plasma concentrations of approximately 80 ng/ml were observed; the half-life of drug removal was approximately 3 hr.

Half-Life↗

Gas chromatography chemical ionization mass spectrometric analysis of diminazene in plasma.

A quantitative and sensitive method was developed for the determination of diminazene in plasma. The assay involves the reduction of diminazene to 4-aminobenzamidine and 4-hydrazinobenzamidine. The latter is further reduced to give an additional mole of 4-aminobenzamidine which is extracted, acetylated and condensed with hexafluoroacetylacetone to form a volatile derivative that is subsequently analyzed by gas chromatography chemical ionization mass spectrometry. 4-Aminobenzylamidine was synthesized and used as an internal standard. The method is reproducible and its sensitivity limit using 1 ml of plasma is 0.1 microgram diminazene ml-1. This sensitivity limit is sufficient to detect plasma levels in cattle following therapeutic doses of the drug.

Amidines↗

Selected ion monitoring assay for meclizine in human plasma.

A method has been developed for the quantification of meclizine in human plasma using gas chromatography mass spectrometry. Pentadeuterated meclizine was synthesized and used as an internal standard. The method gave linear results over a concentration range of 5-200 ng ml-1 and was used to determine plasma levels of meclizine following administration to a male volunteer.

Chromatography, Gas↗

Determination of diminazene in plasma by high-performance liquid chromatography.

An analytical method is described for determining diminazene levels in plasma of cattle. Diminazene is reduced to 4-aminobenzamidine which is extracted and assayed by high-performance ion pair reverse phase partition chromatography. 3,4-Dimethoxybenzamidine served as an internal standard. Diminazene levels as low as 0.1 microgram per ml were detected.

Amidines↗

Metabolic studies on haloperidol and its tetrahydropyridine analog in C57BL/6 mice.

The neuroleptic agent haloperidol (HP) is biotransformed in humans to a pyridinium metabolite, HPP+, that displays neurotoxic properties resembling those of the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-derived neurotoxic pyridinium metabolite MPP+. We report here that HP and its tetrahydropyridine dehydration product 4-(4-chlorophenyl)-1-[4-(4-fluorophenyl)-4-oxobutyl]-1,2,3,6- tetrahydropyridine (HPTP) are metabolized in vivo by the MPTP-susceptible C57BL/6 mouse to several pyridinium metabolites including HPP+ and the 4-(4-chlorophenyl)-1-[4-(4-fluorophenyl)-4-hydroxybutyl]pyridinium species RHPP+, the pyridinium species corresponding to reduced haloperidol (RHP), a major circulating metabolite of HP. Atmospheric pressure ion-spray (API) mass spectral data also suggest the formation of fluorophenyl ring-hydroxylated derivatives of these two pyridinium metabolites. Furthermore, HPLC tracings reveal the presence of HPP+, RHPP+, and two phenolic pyridinium metabolites in brain tissue extracts of HPTP, but not HP, treated mice. The neurotoxic potential of MPTP-type pyridinium species suggests that these metabolites may contribute to some of the neurological disorders observed in humans undergoing chronic HP treatment.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Determination of carbadox-related residues in swine liver by gas chromatography/mass spectrometry with ion trap detection.

The ion trap detector (ITD), in combination with a capillary gas chromatograph and under chemical ionization conditions, offers sufficient sensitivity to determine carbadox-related residues as the methyl ester derivative of quinoxaline-2-carboxylic acid at 3 micrograms/kg or higher in porcine liver. A tetradeuterated internal standard of QME effectively compensates for losses incurred during sample preparation. The method produced mean levels of 3.3 (+/- 0.5), 5.5 (+/- 0.8), and 10.1 (+/- 0.9) micrograms/kg for liver fortified at 3, 5, and 10 micrograms/kg. When applied to analysis of samples containing incurred residues of 14C-carbadox at the low microgram/kg level, results were comparable to those obtained by reverse isotope dilution analysis.

Animals↗