$0pa g 14, This document c o n t a i n s. This i s copy. August DETJBMINATION OF PLUTONIUM IN INWIN URINE. Work Gone By:
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1 c 9of 123c o p i e s - This i s copy August 14, 1947 This document c o n t a i n s DETJBMNATON OF PLUTONUM N NWN URNE Work Gone By: ReDort W r i t t e n Bv: Biochemistry S e c t i o n o f t h e Health Division Wright H Langham $0pa g - 86
2 Abstract A d e t a i l e d account i s given o f t h e c o l l e c t i o n, a s h i n g and analysis o f a 24-hour u r i n e sample Cor plutonium This i s a n account of t h e method c u r r e n t l y i n u s e a t the Los Alamos L a b o r a t o r i e s for t h e d i a g n o s i s ' of, exposure o f personnel_4_q_p7utonium 4 ' s t a t i s t i c a l method f o r e v a l u a t i n g r e s u l t s is a l s o given! DSCLAMER This report was prepared as an account of work sponsored by an agency of the United States Government Neither the United States Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owned rights Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not necessarily constitute or imply its endorsement, recommendation, or favoring by the United States Government or any agency thereof The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States Government or any agency thereof F, *,!'-,,
3 -3- C o l l e o t i o n o f Sample A 2 b h o U r u r i n e sample i s c o l l e c t e d i n a mepr flask \ Extreme, care is t a k e n t o a v o i d contamination of t h e sample during oollection r e t a i n e d in 3 l i t e r Pyrex Grlen- 8: The funnel and f l a s k f o r c o l l e c t i n g t h e sanple a r e s p e c i a l plywood box w i t h a hinged l i d The i n d i v i d u a l on whom t h e t e s t i s t o b e m d e i s i n s t r u c t e d t o remain a t home o r in t h e h o s p i t a l d u r i n g t h e 24rhour p e r i o d over which he is t o c o l l e c t the S a ~ ~ ~ p l e i s i n s t r u c t e d ' t o b a t h e oarefully a n d?enmi0 i n pajamas throughout t h e day He is r e q u e s t e d t o put on w h i t e c o t t o n gloves each time he voida t o minimize t h e p o s s i b i l i t y o f contaminating the sample b y e p i t h e l i a l c e l l s from t h e hands A t each v o i c i n g, t h e l i d t o t h e s p e c i a l sample box i s r a i s e d and t h e u r i n e c o l l e c t e d in such a way t h a t t h e funnel and c o l l e c t i o n M a s k are never touched b y t h e i n d i v i d u a l A f t e r urination t h e l i d o f t h e box i s carefulzy c l o s e d t o p r o t e c t t h e sample from d u s t and contamination s A t the end o f t h e 2 b h o u r o o l l e c t i o n period, t h e sample i s t r a n s p o r t e d - t o t h e l a b o r a t o r y b e f o r e it i s removed from t h e s p e c i a l plyrroqd box 11 B n a l y s i s of a 24-hour Urine Sample A Evaporation and Ashing o f Sample 1 Add d i r e o t l y t o t h e urine samplein t h e o o l l e c t i o n flask about 200 ml oon,ce HNO?, drops n-octyl a l c o h o l and grains of 20 mesh carborundum and p l a c e on a high temperature hot p l a t e c o n t r o l l e d b y a variac
4 2 Begin e v a p o r a t i o n slowly a t low temperature t u r e p r o g r e s s i v e l y a t %lo Raise tempera- minute i n t e r v a l s until danger of bumping and f r o t h i n g i s over and sample i s b o i l i n g vigorously 3 To f a c i l i t a t e e v a p o r a t i o n blow a g e n t l e s t e a d y stream o f f i l t e r e d a i r d i r e c t l y i n t o t h e f l a s k a s soon 4 -Do n o t approach d r y n e s s f T r a n s f e r from f l a s k t o 500 ml p o r c e l a i n c a s s e r o l e and quanti- t a t i v e l y r i n s e f l a s k t w i c e w i t h h o t conc HNO3 cas ser ol e b o i l i n g begins evaporate^ sample u n t i l t h e 1 eve1 of l i q u i d i n f l a s k i s a b o u t 1/2 inch (about 150 ml) 5 at3 Transfer washings t o 6 Cover c a s s e r o l e w i t h speedy-vap c o v e r g l a s s and e v a p o r a t e down a t medium h e a t u n t i l sample b e g i n s t o puff up f u l l y that it does n o t foam and f r o t h over &itch sample c a r e - ' 7 Continue t o e v a p o r a t e sample t o d r y n e s s b y c a r e f u l heating f r e a k t i o n t e n d s t o become t o o v i o l e n t, alternate p e r i o d s o f h e a t i n g and cooling f t h e sample i s heated t o o r a p i d l y it m y i g n i t e a t t h i s p o i n t 8 Take c a s s e r o l e o f f of h o t p l a t e and w h i l e s t i l l hot c a r e f u l l y wash d m s a l t s w i t h conc HNO 9 3 adding t h e a c i d dropwise from a dropper Reheat c a s s e r o l e t o e v a p o r a t e t h e HN03 and r e p e a t s t e p 8 u n t i l s a l t s a r e almost completely w h i t e w i t h t h e exception o f a few aggreg a t e s o f carbon 10 When 8altS over a F i s h e r burner a r e w h i t e remove speedy-vap and i g n i t e c a s s e r o l e, Heat t h e edges o f o a s s e r o l e f i r s t by i n o l i n i n g it on a r i n g and r o t a t i n g it i n t h e fiame o f carbon a r e gone Finally i g n i t e until a l l traoas
5 -5-11 Cool c a s s e r o l e a b o u t 10 minutes and wash walls w i t h a f i n e stream o f 2 N HC The HC1 t r e a t m e n t should b e g i n b e f o r e t h e f u s i o n mass has completely cooled f t h e fused mass is allowed t o s o l i d i f y completely c h i p p i n g of t h e glazed s u r f a c e o f the, c a s s e r o l e may result Algo the sdts go i n t o s o l u t i o n 1 ess r e a d i l y 32 Continue t o wash down walls and speedy-vap w i t h 2 N HC1 u n t i l t o t a l volume o f acid i s a b o u t ml Scrub c a s s e r o l e an2 speedy- uap w i t h policeman and p l a c e c a s s e r o l e back on hot p l a t e a t medium h e a t and r e f l u x u n t i l most o f t h e s a l t s a r e i n s o l u t i o n f s a l t s do n o t go i n t o s o l u t i o n s c r u b s i d e s of c a s s e r o l e w i t h policeman, add more a c i d and The t o t a l volume of s o l u t i o n should n o t exceed 100 m l reheat B \ E x t r a c t i o n and Analyeis 1 Transfer h o t s o l u t i o n and any undissolved s a l t s from c a s s e r o l e t o 240 ml c e n t r i f u g e b o t t l e and c e n t r i f u g e a t 3500 rpm 2 f o r 1 4 minutes C a r e f u l l y d e c a n t s u p e r n a t a n t i n t o second c e n t r i f u g e b o t t l e Wash t h e p r e c i p i t a t e i n t h e f i r s t c e n t r i f u g e b o t t l e w i t h ml o f h o t c o n c e n t r a a d HCl t h a t was heated i n t h e o r i g i n a l c a s s e r o l e S t i r up t h e p r e c i p i t a t e and h e a t c e n t r i f u g e b o t t l e on hot p l a t e f o r few minutes Centri- fuge f o r 10 minutes and t r a n s f e r s u p e r n a t a n t t o second c e n t r i f u g e b o t t l e 'Nash o r i g i n a l c a s s e r o l e and p r e c i p i t a t e i n f i r s t c e n t r i f u g e b o t t l e w i t h m l o f h o t d i s t i l l e d water after c e n t r i f u g i n g f o r 10 minutes 3 T r a n s f e r wash t o second c e n t r i f u g e b o t t l e Discard any remaining p r e c i p i t a t e To t h e a c i d s o l u t i o n (about 125 ml t o t a l volume) add con- c e n t r a t e d NHLOH u n t i l a p e r m n e n t p r e c i p i t a t e forms Add m l o f con-
6 j card t h e supernatant - Dissolvs p r e c i p i t a t e i n nil of hn HC1 Wash d m t h e walls of t h e o e n t r i f u g e B o t t l e d u r i n g t h e a d d i t i o n of t h e a c i d Transfer s o l u t i o n q u a n t i t a t i v e l y t o 2'50 m l p e a r shaped, s e p a r a t o r y funnel Rinse c e n t r i f u g e b o t t l e twioe w i t h 5 m l p o r t i o n s of 4-2N HCl To t h e s o l u t i o n in s e p a r a t o r y funnel add a b o u t 1 mg of f e r r i c i r o n as a s o l u t i o n of FeC13 6 M e u t r a l i e e w i t h c o n c e n t r a t e d NH4OH u s i n g methyl v i o l e t as an indicator ' The end p o i n t should be t h e p o i n t where t h e i n d i c a t o r i s an emerald g r e e n ( b r i g h t green) f a p r e c i p i t a t e should form a d d 2B HC until p r e h i p i t a t e j u s t dissolves 7 Add 2 ml of f r e s h l y prepared aqueous 6% c u p f e r r o n s o l u t i o n Shake s e p a r a t o r y funnel f o r a t l e a s t 1 minute 8 Add 5/4 ml o f CFC13 and e x t r a c t f o r about 1 minute nvert funnel and r e l i e v e p r e s s u r e b e i n g c a r e f u l n o t t o l o s e any of t h e materid Draw o f f CHC13 i n t o 40 ml graduated c o n i c a l c e n t r i f u g e cone 9 Continue e x t r a c t i o n w i t h 3 nil portions o f CHCl3 until ohloroform l a y e r is c o l o r l e s s (about G e x t r a c t i o n s ) 10 Evaporate o f f CHC13 i n a n o i l o r water b a t h a t 6 ~ ~ (avoid superheating) 11 To t h e r e s i d u e i n c e n t r i f u g e cone add 3/4 d o f conc HN03 and 1 ml 72% H C l O 4 Wash down t h e vralls of cone d u r i n g a d d i t i o n Nix w e l l by r o t a t i n g cone 12 P l a c e oone i n a c o n t r o l l e d wax b a t h and g r a d u a l l y i n a r e a s e temperature t o 15OOC 150% Swirl t h e cones o o c a s i o n a l l y u n t i l foaming has subsided continue t o heat a t
7 13 A f t e r foaming has subsided continue t o elevate temperature " to 19b200 A l l o w p e r c h l o r i c a c i d s o l u t i o n to r e f l u x a t t h i s temperature Clean walls of cone by i n c l i n i n g and r o t a t i n g care- f o r a b o u t 1 1/2 hours The volume of t h e a c i d should have been evaporated down t o 1/2 m l fully o r 168s 14 The a c i d s o l u t i o n should b e c o l o r l e s s when cooled down Dilute t o exactly 4 ml d t h d i s t i l l e d water Ad6 2 drops of 2% hydro- xylamine hydrochloride s o l u t i o n and let stand f o r about 1 hour' Add 200 micrograms of hi+++in a b o u t 01 ml of L a ( N 0 ) 15 solution 3 3 Mix Add 1 ml o f conc FiF' and s w i r l t h e cone f o r a few minutes Centrifuge for 10 minutes a t 3800 rpm Decant and Oiscard t h e supernatant n v e r t cone on p i e c e o f f i l t e r paper t o drain of 01 8 HF swirling Wash p r e c i p i t a t e a i t h 2 ml Break up t h e p r e c i p i t a t e and wash dovm walls of cone by C e n t r i f u g e as before Discard s u p e r n a t a n t and invert cone t o dra in 16 T r a n s f e r p r e c i p i t a t e q u a n t i t a t i v e l y t o s t a i n l e s s steel p l a t e T r a n s f e r i s accomplished by s l u r r y i n g t h e p r e c i p i t a t e i n a b o u t 2 drops o f d i s t i l l e d water and drawing t h e slurry up into a c a p i l l a r y s y r i n g e p i p e t t e 17 Evaporate water on h o t p l a t e a t s u f f i o i e n t l y l o w temperature t o avoid s p a t t e r i n g * gnite t h e s t a i n l e s s steel p l a t e l i g h t l y f o r a b o u t 1 minute i n t h e flame o f a F i s h e r burner t o burn o f f o r g a n i c mtter counter 18 Count f o r two one-hour p e r i o d s i n a low baokground a l p h a *
8 9 111 General C o n s i d e r a t i o n s and n t e r p r e t a t i o n of R e s u l t s The a n a l y s i s of hour u r i n e &mples exposed t o plutonium gave a n a v e r a g e b l a n k va7ue o f f r o m persons never c/m There i s a g r e a t p r o b a b i l i t y that t h i s p o s i t i v e b l a n k a c t u a l l y r e p r e s e n t s a naturally occurring c u p f e r r o n e x t r a c t a b l e a l p h a a o t i v i t y o f t h e body The standard p r a c t i c e a t t h i s l a b o r a t o r y i s t o s u b t r a c t a b l a n k v a l u e of 1 c/m from each r e s u l t following formula : The f i n a l r e s u l t -is i n t e r p r e t e d according t o t h e E where PE = probable e r r o r T = 30 minutes - counting time B =mean background of counter (035 c/m) N s mean d e v i a t i o n o f b l a n k ( c/m --an e a r l y value) c/m i n sample ( c o r r for c o u n t e r background and ave blank)
9 -9- T a b l e 1 ; STATSTCAL SGNFCANCE OF RESULTS Probable E r r o r (c/mj with m o b a b i l i t y of Occurrence of a S p e c i f i c Error 1 in 2 1 i n 10 5 i n in eo *54 1 * ! 1 in Z81 3 *93
10 -10- r background and a b l a n k v a l u e of 1 c/m) t h e r e i s one chance in two t h a t it i s an e r r o r b y a s much a s 099 c/m and one chance i n one hundrec t h a t it i s an e r r o r by a s much as 34 c/m A t t h i s ' l a b o r a t o r y 1 microgram o f p l u t o n i m i s c o n s i d e r e d t h e maximum safe body t o l e r a n c e f t h e u r i n a r y e x c r e t i o n r a t e i s t a k e n as 001% of t h e body c o n t e n t per 24-hour period and t h e a c t i v i t y of plutonium t a k e n a s 70,000 c/m/y t i o n of 1 x 10-4 x t h e n a t o l e r a n c e d o s e i s considered t o be t h e exore- 7 x 10-4 = 7 c/m/2bhour sample of u r i n e m e n deter- mined b y t h e c u p f e r r o n procedure and i n t e r p r e t e d b y t h e above formula t h e r e i s one chance i n two that t h e t o l e r a n c e v a l u e i s i n e r r o r by a s much as 096 c/m, as 36 c/m! and one chance i n one hundred t h a t t h e e r r o r i s as g r e a t
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