Dissolution of biological materials combines the simplicity of oxygen-flask combustion with the reproducibility and purity of the final product, and convenience of direct in-vial collection of the sample by the sealed-tube method. It assures quantitative and reproducible recoveries

Huebner, L. G.

Kisieleski, W. E.

English

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Glass To Covar 
Seal - 
Silver Soldered 
Covar To Steel 
Seal 
Stainless Steel 
Adapter 
)-Ring 
tillation Vial 
Trap —78°C 
2OO°C)
p 
Liquid N2 
September 1969	 Brief 69-10412 
•1	 AEC-NASA TECH BRIEF 
AEC-NASA Tech Briefs describe innovations resulting from the research and development program 
of the U.S. AEC or from AEC-NASA interagency efforts. They are issued to encourage commercial 
application. Tech Briefs are published by NASA and may be purchased, at 15 cents each, from 
the Clearinghouse for Federal Scientific and Technical Information, Springfield, Virginia 22151. 
Direct In-Vial Collection for Liquid-Scintillation

Assay of Carbon-14 and Tritium 
To Vacuum Pump 
A
	
B 
Fig. 1. Diagram of the apparatus: A, combustion flask, cold trap, and adapter 
for the scintillation-vial assembly; B, sublimation assembly. 
The problem: 
To develop a better technique for dissolution of 
biological materials for liquid-scintillation studies. 
Earlier methods include dissolution of the materials 
in an organic base such as hyamine hydroxide or 
Nuclear Chicago solvent, the oxygen-flask combus-
tion method, the vacuum-Fine or sealed-tube combus-
tion method, and the in-vial combustion method; all
have certain limitations and have to be modified to 
fit particular requirements. 
The solution: 
A technique for dissolution of biological materials 
that combines the simplicity and facility of the 
oxygen-flask combustion with the reproducibility, 
efficiency, purity of the final product, and con-
venienceof direct in-vial collection of the sample by 
(continued overleaf) 
This document was prepared under the sponsorship of the Atomic	 liability resulting from the use of the information contained in this 
Energy Commission and/or the National Aeronautics and Space document, or warrants that the use of any information, apparatus, 
Administration. Neither the United States Government nor any per-	 method, or process disclosed in this document may not infringe pri-
son acting on behalf of the United States Government assumes any	 vately owned rights.
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the sealed-tube method. The method has these ad-
vantages: 
1. The final product of combustion (H 20 for tritium, 
and CO 2 for 14 C is collected directly into the scin-
tillation vial used for counting, with elimination of 
dilution of the sample and of losses due to transferral. 
2. The final combustion products are pure enough 
for direct gravimetric or volumetric assay. 
3. It eliminates quenching effects due to impuri-
ties or formation of color. 
4. It assures quantitative and reproducible re-
coveries. 
5. Larger samples, weighing 10 to 100 mg dry, 
can be combusted. 
6. Double tracers can be separated with slight 
modification of the apparatus. 
7. It lends itself to rapid routine handling so that 
a reasonable number of samples can be processed in 
1 day. 
With this method various tissues, including blood, 
liver, and eyes, can be combusted quantitatively. 
Samples of nonvolatile organic compounds, such as 
amino acids, carbohydrates, and phosphate deriva-
tives, also have been successfully determined. 
How it's done: 
The apparatus (Fig. I)consists of a 500-ml flask 
fitted with a needle valve, a cold trap with a side arm 
to which a scintillation, vial is attached by way of a 
specially constructed connector and Covar seal, and 
a vacuum pump. Approximately 20 to 30 mg of dried 
tissue is weighed into a gelatin capsule, wrapped in 
black filter paper, and burned in an atmosphere of 
oxygen. The flask is attached to the cold trap, im-
mersed in a mixture of dry ice and alcohol, and ex-
hausted slowly through the trap. Water vapor is 
separated from other gases and condensed in the trap. 
After 5 minutes stopcocks are closed, the flask is 
disconnected, and the trap is removed from the bath
of dry ice and alcohol. The bottom of the scintillation 
vial is immersed in liquid nitrogen, and water is sub-
limed into the scintillation vial. In order to facilitate 
the transfer of water from the trap into the vial, a 
heater is placed around the trap. Sublimation takes 
no longer than 10 minutes before the vial is removed, 
proper scintillation solution is added, and the sample 
is counted in a liquid-scintillation counter. 
By use of four to five trap-vial combinations, the 
process is made continuous, and between 40 and 50 
samples can be processed in 8 hours. The memory 
effect in the trap is less than 0.01%; in the trap-
flask combination: 0.03%. Thus a flask can be reused 
for several samples. 
Reference: 
1. L. G. Huebner and W. E. Kisieleski, in ANL-7278 
(Argonne National Laboratory, Dec. 1968), p. 245. 
Notes: 
1. This information may interest medical or bio-
logical laboratories, or drug or food companies. 
2. Inquiries may be directed to: 
Office of Industrial Cooperation 
Argonne National Laboratory 
9700 South.Cass Avenue 
Argonne, Illinois 60439 
Reference: B69-10412 
Source: L. G. Huebner and W. E. Kisieleski 
Biological & Medical Research Division

(ARG- 10424) 
Patent status: 
Inquiries concerning rights for commercial use of 
this innovation may be made to: 
Mr. George H. Lee, Chief 
Chicago Patent Group 
U.S. Atomic Energy Commission 
Chicago Operations Office 
9800 South Cass Avenue 
Argonne, Illinois 60439
. 
Brief 69-10412
	 Category 03


Direct in-vial collection for liquid-scintillation assay of carbon-14 and tritium

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