ATTACHMENT 13 Contract No. W-7405 , Director edited by J.J. July 23, 1945 Report received: July 25, 1945 VII Excretion Studies Wright Langham The primary interest of our health department is the immediate development of a method of monitoring personnel for internal body contamination with plutonium. The obvious purpose of a monitoring plan is to enable us to retire individuals form further contact with the material before they have absorbed harmful amounts. The execution of such a plan depends on the establishment of a number of factors among which are the following: 1) The development of a method of determining exceedingly small amounts of plutonium in some body fluid or excretment; 2) The establishment of the relationship between the body fluid or excrement and the amount of plutonium contained in the human body; 3) The development of a sampling system which excludes the possibility of external contamination of the sample. This report summarizes our attempts to establish some of the above factors. The urine has been chosen as the source of the sample for study. Method of Sampling and Analysis: Because of the extreme difficulty of detecting small amounts of internal contamination with plutonium, and because of the great possibility of external contamination of the sample, the practice has been to collect 24-hour samples under very rigorous conditions. The subject is directed to stay away from work and preferably away from the site for a 48-hour period preceding the period of collection of the sample to be analyzed. All persons are asked to wear freshly laundered clothing during this preliminary period and to bathe and wash their hands frequently. The subject is asked to report to the hospital at eight o'clock in the morning at the close of the 48-hour preliminary period. He is given hospital clothing and after taking a shower, is admitted to a special room provided for collecting the 24-hour urine sample. He is asked to remain in this room for the entire 24-hour period. It is requested that the subject restrict his fluid intake to one cup or glass of fluid per meal to avoid an abnormally large sample. A hand counter is available in the room and a note is made as to whether or not the individual has a hand count. The subject is instructed to wash his hands each time before he voids and to wear white cotton gloves during voiding, thus preventing epithelial scales of the hands from falling into the flask and contaminating the sample. The voidings are collected in a 2 liter flask which is placed at such a height that it is not necessary for the person to touch the flask or the flask while urinating. When the collection is completed, the subject dresses and leaves the hospital leaving his specimen where it was collected. The sample is picked up and delivered to the laboratory by a member of the group doing the analyses. 2 leave the hospital leaving his specimen where it was collected. The sample is picked up and delivered to the laboratory by a member of the group doing the analyses. The effectiveness of the above method is indicated by the data in Table XIII which gives a comparison of the analyses of samples consisting of two overnight specimens collected in the individuals' house with 24-hour samples collected from the same individuals by the above hospital method. The average counts per minute obtained in the samples collected at home was 20 as compared to 2.2 counts per minute per sample when collected under hospital conditions. The most probable explanation of this great differences is that external contamination was avoided in the latter case. The samples collected in the hospital are analyzed by the following method: The entire 24-hour specimen is evaporated almost to dryness and the ILLEGIBLE using one addition of ILLEGIBLE. HCl and repeated additions of [ILLEGIBLE} and H2O2. The is continued until a almost completely free of organic matter is all that remains. The is taken up in 2 and a complete precipitation carried . The precipitate is dissolved in 2 % HCl, the solution is to a pH 0.3-0.5 and the Pa, plns 1 mg. of ferris iron as a carrier, is extracted into chloroform using . The chloroform is evaporated off and the off with and perchloric acids. The is then carried out of the acid solution with fluoride. The precipitate is transferred to a and counted for 30 minutes in an alpha counter. The data reported in Table XIV give some idea as to the performance of this method when applied to spiked urine samples and to urine solutions. Blank determinations were made on 24 samples of urine from persons never having worked with Pu. These samples ranged in size from 800 to 1200 ml. The average of all blank determinations was 0.5 c/m per sample with a spread of 0-1.2 c/m. Results of Personnel Monitoring: Thirty-six members of the staff were chosen for the first test of the above monitoring method. These people were chosen to represent high, moderate, and low or no exposure groups. The number in each group was too few to give any definite significance to the classification. The results are indicative, however, and are summarized in Table IV. It may be significant that all individuals showing a positive count in the urine had had one or more high nose counts on record since joining the project. A high nose count is recorded against an individual when a moist filter-paper swab inserted into the nostril and rotated shows 50 c/m or greater when counted in an alpha counter. 3 Urinary Excretion of Plutonium by the Human: If urinary excretion values are to be used to establish the actual amount of internal body contamination it is essential to know the relation between the amount of Pu in the human system and that in the urine per 24 hours. On April 10, 1945, an attempt was made to establish this relationship by injecting a human subject intravenously with 4.7% of +4 Pu which was with sodium citrate (0.3% solution) and adjusted to a [ILLEGIBLE] of 6.0.. The subject was an elderly male whose age and general health was such that there is little or no possibility that the injection can have any effect on the normal course of his life. The patient might not have been an ideal subject in that his kidney function may not have been completely normal at the time of injection as indicated by slight and a low urine specific gravity. The [ILLEGIBLE) citrate complex was used in order to produce the maximum [ILLEGIBLE] in the . This presumably would produce an excretion rate comparable to that of a worker having absorbed the material at a slow rate thereby depositing a maximum amount in the where it is probably the most damaging. The results obtained for the first 18 days after injection are presented graphically in Figure 1 by blocking in the per cent of the total injected dose excreted per day. These data show the excretion during the first day was surprisingly low and that the leveling off of the excretion rate was much slower than with rats. The most probable explanation of these observations is that they represent some metabolic abnormality of the subject. It is possible, however, that the stability of the +4 citrate complex is a factor. A blood sample taken 4 hours after the injection showed that about 50 per cent of the injected dose was still in the circulating blood. The calculation, however, was based on the assumption that there had been a complete mixing of the material throughout the total blood volume. A rather favorable excretion rate is indicated by the observation that the leveling off point seems to be about 0.02 per cent instead of 0.01 per cent as observed for rats. The Effect of Size of Dose on Urinary Excretion of 49: A number of fundamental assumptions must be made in regard to the metabolism of Pu if a limited amount of human tracer data are to form the basis of a method of diagnosing internal body contamination. (1) It is necessary to assume that, once absorbed, all valence states and all compounds of Pu are metabolized by the animal organism in essentially the same way. (2) It is necessary to assume that Pu is metabolized in the same way regardless of the route of absorption or administration. (3) It is also necessary to assume that the fraction positive and therefore the fraction excreted is independent of the size of the dose administered or absorbed. Hamilton (CM-2383) has reported a limited amount of information in support of the validity of the first two assumptions. The following experiment was performed to test the validity of the third. 4 Five groups of mature male rats were injected with [ILLEGIBLE] respectively. The material was administered as [ILLEGIBLE] citrate complex in a solution 0.5 per cent with sodium citrate. The pH of the solution was 6.0. The urine and feces were collected daily for five days from each group and analyzed for 49. The results of the urine analyses are given in Table XV1. These data show rather conclusively that the per cent of the total injected dose excreted in the urine of the rat under the above conditions is independent of the size of the dose administered. 5 Table XIII Effect of Method of Collecting Sample on Counts Found in the Urine FOR REFERENCE SEE (11bb02.gif) 6 Table IV Results of Monitoring Site Personnel FOR REFERENCE SEE (11bb03.gif) 7 Fig III - FOR REFERENCE SEE (11bb04.gif) 8