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Biomedical subjects

F Kusu

Publications and source records attributed to F Kusu.

11 recordsLinked to original sources

Determination of p-hydroxymandelic acid enantiomers in urine by high-performance liquid chromatography with electrochemical detection.

High-performance liquid chromatography (HPLC) with electrochemical detection using a chiral ligand-exchange column was developed for the enantioselective determination of p-hydroxymandelic acid (HMA), a metabolite of synephrine, with high sensitivity. A good linear relationship between current ratio and amount was noted for 0.5-500 pmol HMA, with a correlation coefficient of 0.999 for each HMA enantiomer. The relative standard deviation (R.S.D.) was 1.6% at 100 pmol d-HMA and 2.2% at 100 pmol l-HMA. The detection limit of each HMA enantiomer was 0.5 pmol (signal to noise ratio, S/N = 3). By this method, HMA in Citrus unshiu and in urine following the ingestion of C. unshiu was determined. Although no HMA was found in c. unshiu, d- and l-HMA were present in urine after the ingestion of C. unshiu. The time courses of HMA and conjugated synephrine enantiomers excreted in urine following the ingestion of C. unshiu for 24 h could be monitored. This method should prove applicable to the study of synephrine metabolism.

Adult

Determination of acid values of fats and oils by flow injection analysis with electrochemical detection.

A new method using a flow injection system with electrochemical detection was developed to determine acid values of fats and oils. VK3 (2-methyl-1,4-naphthoquinone) solution, i.e., ethanol containing 3 mM VK3 and 38 mM LiClO4, was used as the carrier solution. Flow signals were monitored at -0.33 V vs. Ag/AgCl. For preparation of a sample solution, an oil sample was completely dissolved in VK3 solution, or fatty acids were extracted from the sample into this solution. Aliquots (5 microliters) of the sample solution were injected into the flow injection system. Acid values were determined based on flow signals for 14 samples and the results were found to be consistent with those by potentiometric titration. Relative standard deviation was less than 2%. Samples were processed at the rate of 60 h-1. The stability of fish and cod liver oils was followed by measuring acid values for 8 weeks. This method proved to be a simple and rapid means for acid value determination.

Acids

Selective determination of chloride and bromide ions in serum by cyclic voltammetry.

Voltammetric determination of chloride and bromide ions in serum was made using a one-body type Ag electrode, whose surface was covered with a dialysis membrane for preventing interference from proteins. This determination was based on measurement of the charge of the reduction wave of silver halide formed on the Ag electrode surface in a halide ion solution during a cathodic potential sweep. In this method, depending on the potential sweep range, the charge of the reduction wave of AgI, AgI + AgBr, or AgI + AgBr + AgCl can be measured. Here, the method was used to determine the chloride and bromide ions in artificial serum. Linear concentration ranges were 1.0 x 10(-2)-2.4 x 10(-1) M for chloride ions and 5.0 x 10(-3)-1.8 x 10(-2) M for bromide ions. The correlation coefficient was 0.999 in each case. The relative standard deviation for chloride ion (1.0 x 10(-1) M) was 1.9% (n = 10), and that for bromide ion (1.5 x 10(-2) M) was 2.5% (n = 10). Bromide ions at a relatively high concentration as the component of serum had no effect on the determination of the chloride ion in the serum. This method is thus useful for the selective determination of chloride and bromide ions in serum.

Blood Substitutes

Determination of synephrine enantiomers in food and conjugated synephrine in urine by high-performance liquid chromatography with electrochemical detection.

Determination of synephrine enantiomers was made with HPLC with electrochemical detection using a chiral ligand-exchange column (Sumichiral OA-6000). The calibration curve for each enantiomer showed good linearity (r = 0.999) at 1.0-500 pmol injected with a detection limit of 1.0 pmol (signal-to-noise ratio (S/N) = 3). Relative standard deviation (RSD) was 1.8% at 50 pmol d-synephrine and 1.6% at 50 pmol l-synephrine. The contents of synephrine enantiomer in food such as Citrus unshiu fruit, orange juice, and marmalade were determined. The present method was also used to determine conjugated synephrine enantiomers in urine following the ingestion of C. unshiu pulp. l-Synephrine in fruit was converted to the conjugated form of synephrine, and l-synephrine underwent chiral inversion to d-synephrine in vivo.

Adult

Determination of barnidipine in human serum and dog plasma by HPLC with electrochemical detection.

Barnidipine is a 1,4-dihydropyridine calcium antagonist. HPLC was conducted on a polybutadiene coated alumina column using an alkaline mobile phase and an electrochemical detector to determine the content of this drug in serum and plasma. A good linear relationship between barnidipine concentration and peak height was found in 5-500 ng/ml with a correlation coefficient of 0.998. The detection limit was 1 ng/ml. The within-day and day-to-day variations were examined for control human serum. Relative standard deviation of within-day assay for serum spiked with 10 ng/ml barnidipine.HCl was 6.9% and the recovery was 104%. A pharmacokinetic study was made in which the time course of barnidipine in dog plasma was followed.

Animals

Fluorometric determination of pseudocholinesterase activity in postmortem blood samples.

A fluorometric assay using 3-(p-hydroxyphenyl) propionic acid (HPPA) was conducted to determine the activity of pseudocholinesterase (ChE) [Enzyme Commission (EC) No. 3.1.1.8] in postmortem blood samples so as to test for organophosphate poisoning. By the enzymatic reaction of ChE, its substrate, benzoylcholine, produces choline, which is oxidized by choline oxidase to generate hydrogen peroxide. HPPA is oxidized by hydrogen peroxide and peroxidase to become the fluorogenic dimer whose concentration is measured fluorometrically at an excitation emission wavelength of 320 nm and an elimination emission wavelength of 404 nm. The selectivity and sensitivity of the present method were found to be superior to those of conventional pH and spectrophotometric methods.

Butyrylcholinesterase

Differential pulse polarographic determination of plasma menadione.

A differential pulse polarographic assay for plasma vitamin K3 (menadione) has been developed. Details of the assay are (i) lipid-soluble material is extracted from plasma into ether by the method of Bjornsson et al. [(1978) Thromb. Haemostas. 2, 466-473]; (ii) ether is evaporated under nitrogen and the residue is dissolved in the supporting electrolyte, methanol: 0.2 M borate buffer (9:1), pH 6.8; (iii) current height is measured at -0.32 V vs SCE on the differential pulse polarogram. The lower sensitivity limit of this technique after addition of standard vitamin K3 to plasma is 0.3 microM; the calibration curve is linear from 0.6 through 10 microM. Two patients treated with a single dose of menadiol sodium diphosphate, 20 mg/M2 i.m., achieved measurable plasma vitamin K3 levels at 0.5 to 1.0 h ranging between 0.5 (0.08 micrograms/ml) and 2 microM (0.3 micrograms/ml).

Humans