Masaharu Horiguchi

Determination of prednisone and prednisolone in human serum by high-performance liquid chromatography —especially on impaired conversion of corticosteroids in patients with chronic liver disease

A reliable and rapid method is described for the determination of prednisone and prednisolone in human serum by high-performance liquid chromatography, using a Zorbax-SIL column with dichloromethane-ethanol (92.5:7.5) as eluent, with UV detection at 254 nm. Metabolites and endogenous hydrocortisone did not interfere with the determination of prednisone and prednisolone. The alteration of corticosteroid concentrations in serum from patients with chronic liver diseases was studied following a single oral administration of prednisone or prednisolone (30 mg). The proposed method showed good separation of several corticosteroids and was time-saving, suitable and reliable for the routine analysis of corticosteroids in human serum.

Lactate Dehydrogenase Isozyme Patterns of Mammalian Livers

Liver lactate dehydrogenase (LDH) isozyme compositions were studied in mammals of 17 species (newly studied 12 species and previously studied 5 species, by new samples obtained). Results are summarized as follows:1. “Carnivora type, ” in which LDH-5 is the most dominant, and “Artiodactyla type, ” in which LDH-1 is the most dominant, are observed.2. Among mammals of Artiodactyla, those of a group of deer (Cervidae) is different from those of a group of ox (Bovidae). Proper liver LDH isozyme composition is observed in not only order but also in family.3. Proper pattern is also oderved in the livers of Ass and Malay tapir, belonging to Perissodactyla, and in Japanese monkey, belonging to Primates.4. That of Damma wallaby is also studied.

Studies on the heat stability of alkaline phosphatase in serum

It has been actively studied in many laboratory to differentiate alkaline phosphatase isoenzymes in serum by using electrophoretic techniques, while it also became interesting problem to separate the several types of alkaline phosphatase by heating and chemical inhibitors. Eight cases (such as 3 ofhepatoma, 1 of gastric cancer, 1 &pancreatic cancer, 1 of uterus cancer, 1 of congestive heart failure and 1 of cholangiolitic hepatitis) with relatively heat-stable alkaline phosphatase which had more than 40~ of activity after heating at 56~ for 15min. are observed in 250 cases. Seven cases out of them were died within 2 months and it seems that presence of these relatively heat-stable alkaline phosphatase is a valuable fact for determining prognosis of the patients with severe disease. Serum from 22 cases who had malignancy and died showed remarkable changes of nature of alkaline phosphatase against several inhibition techniques, compared with normal serum. These serum alkaline phosphatase were quite sensitive against inhibition by EDTA, while were relatively stable against heating. Therefore, remarkable difference in ratio of EDTA stable activity/heatstable activity (56~ 15 rain.) was observed and it was 0.47 in these cases, compared with 1.10 in normal subjects and 1.57 in liver cirrhosis. In eight cases which were mentioned above, showed quite low value (0.19) in this ratio. It seems that this ratio is also useful tool for determining the prognosis of patients with malignancy. It is quite difficult question why these characteristic alkaline phosphatase appear in serum of the patients with malignancy or severe disease. More studies are required to dissolve this problem. It was quite interesting data which bile in gallbladder had heat-stable alkaline phosphatase, although liver enzyme was sensitive against heating.