A highly sensitive, specific method for the determination of serum copper was described, for which water-soluble porphyrin as chelating reagent firstly has been used. The molor absorptivity of porphyrin (TMPyP; εeffective=2.3×10⁵) is twenty fold higher than that of bathocuproine. Reducing reagent (CySH) was required not only for the formation of TCA deproteinization system but also for the promotion of the chelating of TMPyP-Cu.
The serum sample (0.2ml) was deproteinized with CySH-HCI-TCA reagent and the supe-rnatant was mixed TMPyP buffer at pH 4.7 for 5 min, then add H2SO4 and mix. Measure the absorbance of the Blank (B), Test (T), and 200μg Cu/100ml as calculated on followed formula standard (S) at 447nm.
(B-T)/(B-S)×200=Serum copper (μg/100ml).
The other serum components (ex. iron, zinc and ascorbic acid) were not interfered.
Reproducibility in the serum for which coPPer of 129.3μg/100ml was±2.865 and 2.22% of C. V. The correlation coefficent of the present method to bathocuproine method was 0.996.

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A new method of radioimmunoassay for canine myoglobin was devised. Chan- ges in myoglobin levels in serum and myocardium of dogs were followed with this technique after acute myocardial infarction was produced by ligation of left anterior descending branch of the coronary artery at a proximal site. The amount of in farction was determined by the method of Hackel after autopsy and the myocardium was devided into 3 parts: intact, border, and infarction zones.
Serum myoglobin levels in normal dogs ranged from 18 to 97 ng/ml (mean 48.6±35.1 ng/ml). The serum level elevated in 15 minutes, reached a peak 5 to 8 hours after ligation and then started to decrease gradually. The maximum levels of serum myoglobin determined in 8 dogs showed a good correlation to the weights of infarction (r=0.731). The myoglobin conentration in the infarction zone (2.74±0.58mg/g wet weight) was significantly (p<0.01) lower than that in the intact zone (3.63±0.75mg/g wet weight). The myoglobin content in the border zone was also reduced, though not so significantly.
It was concluded that serum myoglobin level appears to be a useful indicator in deter- mining the severity of acute myocardial infarction.

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It is well known that the concentration of blood ammonia is elevated with standing after venipuncture, and some of it's mechanism are studied in this paper. Increased ammonia with whole blood samples is observed by about 20% per 60min at 4°C and 50% per 60min at 37°C although the concentration of ammonia in separated plasma does not change with time. The production of ammonia with three different samples, such as, whole red cell, contents of red cell without membrane and crude red cell membrane obtaind from freezing-thawing red cells, was examined and it is found that the ammonia production arised in the red cells. Rappid ammonia production are observed when AMP, ADP and ATP are added to the solution of freezing-thawing red cells, and the concentrations of AMP, ADP and ATP in blood also changes togather with the rappid ammonia production in blood. As AMP deaminase (EC 3.5. 4.6) exists in high activity (175U/L) in the red cell, AMP deaminase is considered to be widely concerned in the ammonia production in the whole blood samples.

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In order to obtain the information of individual differences in drug-metabolism, oxidation and conjugation of aminopyrine in normal volunteers and patients were studied in our preceding paper. This paper describes studies on metabolisms of sulpyrine, isoniazid, barbital and acetaminophen. In the group of low acetylation of aminopyrine, concentrations of acetylated metaboltes of sulpyrine and isoniazid in urine were similarly low, and other conjugated metabolites, glucuronides and sulphates, were excreted in normal ranges.
In normal volunteers, concurrent administration of sulpyrine, isoniazid, barbital and acetaminophen, 100mg respectively, had no effect on metabolism of each other. In patients, acetylation and other conjugations were depressed. The depression was more marked in patients administered these 4 drugs with therapeutic drugs than in patients without therapeutic drugs for 24 hours. The depression was accompanied with increased excretion of the metabolites before conjugation. Metabolism and excretion of drugs were affected by concurrent administration of many drugs and medication for a long time. These results must be considered in efficacy and toxicity of therapeutic drugs to patients.
The gas chromatographic methods modified by us allowed determination of each metabolite of the above 4 drugs administered concurrently, and it is applicable to a test for the drug-metabolism of patients.

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An enzymic method is described for determination of the ratios of esterified cholesterol to total cholesterol in serum lipoproteins.
Human serum lipoproteins were separated by electrophoresis using an agarose gel plate with two sample troughs. After electrophoresis, the gel plate was cut in two parts, and these were separately stained by overlaying commercially available enzymic reagents for determination of total cholesterol and free cholesterol respectively, in order to recognize the positions of lipoproteins in the gels. Thereafter, each lipoprotein area was cut out of the gels and stained in tubeswhich contained the corresponding enzymic reagents.
After staining, the solutions in tubes were analyzed by photometer and the distribution ratios of these cholesterols to each lipoprotein were calculated from the ratios Chol of each lipoprotein to total lipoproteins in optical density. Then, the amount of T. Chol, F. and also the ratios of esterified cholesterol to total cholesterol in each lipoprotein were calculated by multiplying of these distribution ratios with these cholesterol concentrations in serum.
This method needs no special instrument and technique except those in conventional examination, so that it is simple to perform. It has sufficient reproducibility and sensitivity.

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A fundamental study on determinations of minute nicotine and its metabolites has been performed with gas chromatography equipped with high sensitive nitrogen detector.
Hitachi flame thermionic detector (FTD) is very convenient and has a much more specific sensitivity to nicotine when a hydrogen flow rate is reduced. For a stationary liquid phase of column packings, 10% Apiezon L+10% KOH is recommended. When hydrogen flow rate was reduced under 2ml min. a hydrocarbon peak and solvent peak were almost disappeared and a base line became very stable and less noisy. Programing analysis at the temperature from 190°C to 240°C was possible even with a single column.
The coefficient of variation (C. V.) was under 5% in the internal standard method, and the detection limit of this method was about 100pg. as for nicotine.

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A case of macroamylasemia due to the complex of salivary amylase and IgA was studied. Macroamylase and salivary amylase from the patient were inactivated similarly by heat treatment at 50°C. However, serum amylase of normal size and urinary amylase from the patient, both of which were determined as pancreatic isozyme by electrophoresis, were stable at 50°C for 20min. Normal size serum amylase and urinary amylase from the patient was more resistant to EDTA than macroamylase and salivary amylase from the patient.
Heat treatment of amylase after separation by gel filtration can clearly distinguish human amylase isozyme.

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It is well known that the aromatic amines, such as sulfone amide, circulate in the body fluid as free and bound forms. In this report, we investigated the metabolic change of p-amino hippuric acid (PAH) after intravenous injection. Among the patients examined, no significant difference of the ratio of free to bound forms could be found in each blood and urine of the examinees. Comparing PAH clearance value of before and after hydrolyzation the regression line, y=0.87x+29.(r=0.97) was obtained A slightly lower value of PAH clearance was obtained after hydrolyzation. According to the result of thin layer chromatography, it is strongly sugestegd that the bound form of PAH is acetylPAH. From the present study it is suggested that when calculating PAH clearance, one should have an attention on the presence of bound form PAH.

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