Toyokazu Ohki

Gas chromatographic—mass spectrometric analysis of polyols in urine and serum of uremic patients : Identification of new deoxyalditols and inositol isomers

A gas chromatographic--mass spectrometric method was applied to a study of polyols in urine and serum of normal subjects and uremic patients. 4-Deoxythreitol, 4-deoxyerythritol, 5-deoxyxylitol, 5-deoxyarabitol, 2-deoxyribitol, 6-deoxymannitol, 6-deoxygalactitol, neoinositol and chiroinositol were identified in normal urine as well as in uremic urine for the first time. In uremia the urinary excretion of myoinositol and chiroinositol was significantly increased. The serum levels of myoinositol, chiroinositol and scylloinositol were increased in the uremic patients, whereas the serum level of 1-deoxyglucose (1,5-anhydroglucitol) was significantly decreased in the uremic patients as compared with the normal subjects. These findings suggest the altered metabolism of chiroinositol and 1-deoxyglucose in the uremic state.

Pattern of aliphatic dicarboxylic acids in uremic serum including a new organic acid, 2,4-dimethyladipic acid

(1) 2,4-Dimethyladipic acid was first identified in normal human urine using gas chromatography-mass spectrometry. Urinary excretion of 2,4-dimethyladipic acid in 7 healthy adults ranged from 4.9 mumol to 14 mumol per 24 h. (2) Succinic acid, adipic acid, 3-methyladipic acid, 2,4-dimethyladipic acid, pimelic acid and azelaic acid were identified in the ultrafiltrate of the blood obtained from a chronic uremic patient using a hemodialyzer. (3) Levels of succinic acid, adipic acid, 3-methyladipic acid, 2,4-dimethyladipic acid, pimelic acid and azelaic acid in uremic serum were determined using a mass fragmentographic technique. Concentration of succinic acid in uremic serum was comparable to that in normal serum, whereas concentrations of adipic acid, 3-methyladipic acid, 2,4-dimethyladipic acid, pimelic acid and azelaic acid were highly elevated in uremic serum.

Gas chromatographic-mass spectrometric profile of organic acids in urine and serum of diabetic ketotic patients.

The organic acids in the urine and serum of diabetic patients with ketoacidosis and disturbance of consciousness were studied using acidification, extraction, evaporation, methoxime formation and trimethylsilylation, gas chromatographic separation and mass spectrometric identification procedures. The organic acid profile of 1 ml of serum ultrafiltrate was obtained with good separation using a gas chromatograph equipped with a glass capillary column and a splitless injector. 5-Hydroxyhexanoic acid and 3-hydroxyvaleric acid were identified for the first time in the urine of diabetic patients with ketoacidosis. Urinary excretion and serum concentrations of 2,3-dideoxypentonic acid were increased in diabetic patients.

Profiling of uremic ultrafiltrate using high resolution gas chromatography-mass spectrometry - identification of 6 polyphenols.

An analytical method for separation and identification of compounds in uremic ultrafiltrate has been developed using a glass capillary column gas chromatography-mass spectrometry-computer system. The ultrafiltrate samples of blood were obtained during hemodialysis treatment of chronic uremic patients and non-uremic patients using the extracorporeal ultrafiltration method. Sample preparation consisted of acidification, extraction, evaporation, and trimethylsilylation. Many compounds were newly identified in the uremic ultrafiltrate by electron impact ionization, chemical ionization, and high resolution mass spectrometry. Six toxic polyphenols, namely, catechol, resorcinol, hydroquinone, 2-methoxyresorcinol, 3-methoxycatechol, and methoxyhydroquinone were first detected in the uremic ultrafiltrate.

Gas chromatographic-mass spectrometric analysis of organic acids in renal tissue biopsy: identification of 4-hydroxybutyric acid and 4-hydroxy-2-butenoic acid.

The organic acids in renal tissue biopsy (0.5-1 mg) obtained from chronic glomerulonephritis patients were analyzed capillary column gas chromatography--mass spectrometry. Some twenty compounds were identified in the renal tissue. The organic acid profile of renal tissue showed a marked difference from those of urine and serum. In particular, 4-hydroxybutyric acid and 4-hydroxy-2-butenoic acid, which are usually undetectable in urine and serum, were detected for the first time in renal tissue in considerably large amounts.

Profiling of organic acids and polyols in nerves of uraemic and non-uraemic patients.

Organic acids, polyols and lipid-bound polyols in the cauda equina nerves of uraemic patients and non-uraemic patients were analysed with high-resolution gas chromatography-mass spectrometry. In the uraemic nervous tissue, the concentrations of myoinositol and 4-hydroxyphenylacetic acid were increased. Levulinic acid was first detected in the nervous tissue as a normal component. 1-Deoxyglucose and free and lipid phosphatide scylloinositol were detected in the nervous tissue as normal components.

Amine metabolite profile of normal and uremic urine using gas chromatography--mass spectrometry.

A method for the simultaneous analysis of phenolic amines and aliphatic amines in human urine is described. The amine metabolites in urine were extracted using Dowex 50W-X8 cationic resin, derivatized and analyzed by a gas chromatographic--mass spectrometric--computer system. The amine metabolites profile of 5 ml of urine was obtained with good gas chromatographic separation. The gas chromatographic method described here separates urinary phenolic amines, di- and polyamines and methylguanidine in a single chromatographic separation. The urinary levels of methylguanidine, putrescine, cadaverine, spermidine, p-tyramine, dopamine, and 3-methoxytyramine were quantitated by using a mass spectrometric technique. In uremic patients, only the urinary excretion of methylguanidine was increased in comparison with normal subjects, although the urinary excretion of other amines was decreased in uremic patients.

Identification of some abnormal metabolites in psoriatic nail using gas chromatography—mass spectrometry

A gas chromatographic--mass spectrometric analysis was used to separate and identify abnormal compounds in the nail of psoriatic patients. The nail was extracted with heated ethanol, and the extract was analyzed with and without trimethylsilylation. Tetradecanoic acid octadecyl ester, hexadecanoic acid octadecyl ester and octadecanoic acid octadecyl ester were first identified in the psoriatic nail, but were not detected in normal nail.

Ischemic change of organic acids in kidney.

Organic acids in rabbit renal tissue biopsy were analyzed by capillary column gas chromatography--mas s spectrometry. The change of these organic acids under ischemic conditions was determined over 60 min after clamping the renal artery and vein. The results showed that lactic acid, glycolic acid, 2-hydroxybutyric acid, 3-hydroxypropionic acid, 2-methyl-glyceric acid, glyceric acid and malic acid increased at 4 and 6 min after clamping, but then decreased at 15 min. Glycerol increased 2 min after clamping and then decreased. However, 3-deoxyaldonic acids of 3-deoxytetronic acid, 3-deoxy-2-C-hydroxymethyltetronic acid and 3-deoxypentonic acid decreased in the renal tissue biopsy from 2 min after clamping.

Gas chromatographic-mass spectrometric analysis of amine metabolites in urine.

An analytical method for the separation and identification of amine metabolites in human urine has been developed using a gas chromatograph-mass spectrometer-computer system. The sample preparation consists of organic solvent extraction, adsorption on a cation-exchange resin, evaporation, and acylation. N-3-Hydroxypropyl-1, 4-diaminobutane was identified for the first time in urine. Methylguanidine, guanidine, putrescine, cadaverine, p-tyramine, 3-methoxytyramine, spermidine, and spermine were detected in normal urine with good gas chromatographic separation.