The hypothesis of the existence of a phase "oxygen" of offbalance D led us to study the appearance in urine of products resulting from abnormal oxidation. We were especially interested in the existence of substances having peroxide properties. We found that addition of sulfuric acid to urine of certain subjects induced appearance of indigotin and indigo rubin. In order to investigate the reaction, we have utilized the solubility of indigotin and indigo rubin in neutral solvents. Through their extraction it appears possible first to prevent their transformation in colorless isatin and, second, to evaluate the relative amounts when they appear during the reaction. To 4 cc. of urine, one centimeter of toluene was added. After shaking the mixture, 1 cc. of pure sulfuric acid was added and the mixture was immediately shaken again. When the mixture was allowed to stand, the toluene separated and its color, blue or violet, indicated the presence and also the relative amounts of indigotin and indigo rubin.

Another method used to detect peroxides was the acidification of urine followed by addition of potassium iodide. For iodometric evalution, starch solution was added. The amount of iodine liberated could be determined titrimetrically.

Fig. 232. The urinary oxidoreduction values in a case of cancer of the colon with abdominal metastases. The values remain the whole time above 100 seconds, corresponding to the pattern present in the offbalance type D.

The form in which peroxides are present in the urine is not clearly established. Although the distillation of the urine gives peroxides in the first distillate, these are not in the form of hydrogen peroxide, since catalase does not induce their disappearance. The values obtained with both methods, sulfuric acid and iodometric, are relatively parallel. The sulfuric acid method, however, produced a higher percentage of positive results.

The presence of slight amounts of peroxide in the urine has been found in about 3% of normal subjects. In contrast, we found peroxide in the urine of 87% of a group of 27 schizophrenics studied through daily analyses over a period of three years. In some of the subjects, throughout the entire three year period with more than 1,000 analyses, not a single negative reaction was seen. (Fig. 233) (221, 222)

We have also found positive reactions during streptococcic infection, erysipelas or tonsillitis. In radiation sickness the reaction is positive especially when tissue lesions are manifest such as mucositis or epidermitis. In general, treatment with selenium has given a relatively high proportion of positive results, especially at the beginning of treatment. While positive reaction appeared to be consistent with a favorable evolution of tumors, an extremely intensive reaction appeared related to a bad prognosis. The following observation is characteristic.

Fig. 233. The reaction for peroxides remains consistently positive in the urine of a schizophrenic in daily analyses during a period of 3 years. (Part of the curve.).

Mrs. N. C, 28 years old, with Hodgkin's disease, had received three treatments of teleradiotherapy, with the general condition completely unchanged. The patient presented an extremely intensive urine peroxide reaction. Dilution of the urine to 1 /50 still showed a marked blue color after treatment with sulfuric acid and toluene. We informed the attending physician about the finding and advised the discontinuance, at least for the moment, of the treatment. The perfect general condition of the patient induced the radiologist to disregard our advice. A new treatment of teleradiotherapy was administered. Three hours later the patient, who only that morning had been shopping, went into a shock and died during the night.

We want to emphasize, however, the correlation between the lack of peroxides in the urine and a poor prognosis during radiotherapy. The cases in which a positive reaction disappeared, were always followed by a change for the worse in the general condition. The persistence of this lack of peroxides was seen in the cases with a rapid lethal termination.