THE DETERMINATION OF POTASSIUM IN BLOOD

SERUM.*
BY F. H. L. TAYLOR.
WITH THE TECHNICAL ASSISTANCE OF ELEANOR R. SHEA.
(From the Thorndike Memorial Laboratory, Boston City Hospital, and the
Department of Medicine, Harvard Medical School, Boston.)
(Received for publication, March 7, 1930.)

INTRODUCTION.

The existing methods (l-3) for the determination of potas-

sium by its precipitation as the cobalti-nitrite, and subsequent use
of the Ilosvay (4) modification of the familiar diazotization reac-
tion of Greiss, have been modified to increase the speed and
accuracy.
This object has been accomplished by t,he introduction of certain
changes in the Briggs-Doisy-Bell procedure. These changes may
be briefly stated as follows: (1) the introduction of the more
efficient protein precipitant tungstic acid; (2) the avoiding of trans-
fers and filtration, and the use of a new method of evaporation;
(3) washing the precipitate of potassium cobalti-nitrite with 30
per cent alcohol, in which it is lesssoluble than in sodium chloride;
(4) the use of 10 per cent acetic acid to prevent flocculation during
diazotization; (5) the subjection of the standard potassium solution
to the sametreatment as the unknown, as suggested by Domogalla
(5), with its attendant accuracy as previously shown by the
author (6).
Reagents.
Xtandard Potassium Solutions (S).-Dissolve 382 mg. of purest
potassium chloride in water and make up to 1 liter. 100 ml. of this
solution contain 20 mg. of potassium. This is approximately the
potassium concent,ration of normal blood serum.
* The expense for this work was defrayed, in part, by a gift from the
Smith, Kline and French Laboratories, Philadelphia.
27

This is an Open Access article under the CC BY license.

28 Determination of Potassium in Serum
Doisy-Bell Cobalti-Nitrite Reagent (S).-Dissolve 220 gm. of
potassium-free sodium nitrite in 400 ml. of distilled water. Dis-
solve 113 gm. of cobalt acetate in 300 ml. of distilled water. Mix
thoroughly and add 100 ml. of glacial acetic acid. Pass a current
of air through the solution for several hours, until the evolution of
nitric oxide fumes ceases. Filter, stopper, and place in the ice
box. It is suggested that the stock bottle remain in the ice box,
small quantities being removed as required.
Sulfanilic Acid Solution.-A 0.5 per cent solution in 30 per cent
acetic acid.
a-Naphthylamine Solution.-A 0.5 per cent solution in 30 per
cent acetic acid.
Alcohol.-95 per cent; 30 per cent.
Sodium Hydroxide.-0.1 N.
Acetic Acid.-10 per cent.

Procedure.
Preparation of Blood Filtrate.-To 2 ml. of fresh blood serum add
14 ml. of distilled water, 2 ml. of 5 per cent sodium tungstate solu-
tion (introduced accurately with a 2 ml. pipette), and 2 ml. of 0.3
N sulfuric acid. Treat 2 ml. of the standard potassium solution
in a like manner. The precipitation is carried out in small Erlen-
meyer flasks. Shake the contents of the flasks and allow to stand
for 10 minutes. Filter in dry, ashless (Whatman No. 50) filter
papers; 5 ml. of the filtrate are used for each analysis.
Evaporation of Filtrate.-Transfer 5 ml. of each protein-free
filtrate to separate, 15 ml. matched, graduated centrifuge tubes.
Arrange each tube as shown in Fig. 1. The capillary tube is so
arranged that the opening is 1 cm. above the level of the liquid in
the tube. It is drawn out so that, by gentle suction, a partial
vacuum is maintained, and at the same time the current of air
enters with sufficient force to agitate the contents of the tube.
The current of air is adjusted by regulating the vacuum so that
no bumping occurs. The tubes are hung in a large beaker of
distilled water, kept at the boiling point by means of an electric
hot plate. The use of distilled water keeps the contents of the
water bath visible at all times. This type of water bath enables
one to observe the process of evaporation and to adjust the vacuum
as required.
 F. H. L. Taylor 29

Evaporation is continued to dryness, which requires about 30

minutes.
Precipitation.-To the residue obtained from the above evapora-
tion add 1 ml. of distilled water, dissolving the residue with heat-
ing, if necessary. Add 1 ml. of 95 per cent redistilled alcohol and
mix thoroughly by shaking. The precipitation is carried out by
the addition of 1 ml. of Doisy-Bell cobalti-nitrite reagent. After

FIG. 1. Evaporation apparatus. P. C. = screw clamp; R. T. = rubber

tubing; R. S. = rubber stopper; C. T. = centrifuge tube; A. = absorbent
cotton; C. = capillary tube.

thoroughly mixing the contents, the tubes are set aside for 2 hours,
after which they are centrifuged at 1800 to 2000 revolutions per
minute for 10 minutes.
Washing.-The supernatant liquid above the precipitate is
rapidly decanted. Add 3 ml. of 30 per cent alcohol, a small
amount at a time, thoroughly suspending the precipitate on each
addition. Centrifuge rapidly for 5 minutes and decant off the
30 Determination of Potassium in Serum

alcohol. Five such washings are required. The use of alcohol

solutions greater than 30 per cent must be avoided, since they
cause precipitation of some sodium cobalti-nitrite.
After draining from the final washing with alcohol dissolve the
precipitate in 0.1 N sodium hydroxide and transfer to 100 ml.
volumetric flasks, using 20 ml. of the sodium hydroxide for the
combined operations of washing and transferring. The sodium
hydroxide is added in four 5 ml. portions. The actual procedure
for this step has been found to be so important that it is given here
in detail. A few drops of the first 5 ml. portion of 0.1 N sodium
hydroxide are added to the precipitate which is completely sus-
pended by tapping the tubes against the palm of the hand. The
remainder of the first portion is added, 1 ml. at a time, the pre-
cipitate being kept suspended. Most of the precipitate is dis-
solved in the course of this treatment. This portion is then
rapidly decanted into 100 ml. volumetric flasks. The second
5 ml. of sodium hydroxide are added in the same manner. At the
conclusion of the transfer no trace of precipitate should be present
in the tubes. The flasks are then thoroughly shaken and the
tubes washed into them by a stream of water from a wash bottle,
with 50 ml. of water. After being shaken thoroughly again, so
that all the precipitate is dissolved, the contents of the flasks are
diluted to the 100 ml. mark. The flasks should be kept tightly
stoppered with ground glass stoppers to prevent oxidation of the
dissolved nitrite. The flasks are then set asidefor 1 hour.
Diasotixation.-Transfer 5 ml. of the sodium hydroxide solution
from each of the 100 ml. flasks, by pipettes, to 50 ml. volumetric
flasks and dilute to 4 times their volume with 10 per cent acetic
acid. Add 2 ml. of sulfanilic acid and 1 ml. of oc-naphthylamine
solution and again shake thoroughly. Dilute to the mark with
10 per cent acetic acid and mix by inverting the flasks several
times. Allow the color to develop for 10 minutes and compare
in the calorimeter. The standard solution is set at 20, and with
a potassium solution containing originally 20 mg. per cent of po-
tassium the calculation is:
Standard
~ X 20 = mg. per cent potassium
Reading
 F. H. L. Taylor 31

TABLE I.
Potassium Values of Serum of Some Normal and Pathological Individuals.

Patient. Date. Diagnosis. EL per 100 cc. blood.

_ -
19dQ mg.

E. S. Nov. 22 Normal. 22.0

I‘ “ 25 “ 20.0
“ I‘ 25 “ 19.9
“ 24.0
A. L. “ 24
‘I
A. E. C. “ 26 22.2
1930
“ 22.2
I. N. Jan. 4
“ “
M. B. 4 21.9
19B9
A. S. Nov. 25 Infectious arthritis. 13.6
“ L‘ 25 “ “ 13.4
M. R. “ 26 “ ‘I 17.7
“ ‘I 17.0
s. w. “ 30
F. L. Dec. 2 Pernicious anemia. 25.0
Fo. “ 2 “ ‘I 21.6
I‘ 2 “ “ 28.5
Sul.
“ 2
zu. Myxedema. 17.4
cu. “ 2 Pernicious anemia. 25.2
Wa. Nov. 30 ‘I “ 26.6
“ 2 “ “ 23.6
Wi.
“ “ 17.0
Jo. “ 30
- --

TABLE II.
Potassium in Standard Solutions.

K present. K found.* EWX-.

mg. per 100 ml. mg. per 100 ml. per cmt
30 30 zto.0
30 30.7 +2.3
20 20.3 $1.5
20 19.7 -1.5
10 10.2 +2
10 10.0 zko.0

* Standard, 20 mg. per 100 ml.

32 Determination of Potassium in Serum

Experimental Results.
Experimental results showing the range of potassium values in
normal and certain pathological conditions are given in Table I.
Table II shows the amount of potassium obtained from various
concentrations of potassium in aqueous solution. The accuracy
and duplication have been found to be high. The recoveries ob-
tained from standard solutions were within 3 per cent of the true
value.
The method is equally applicable to whole blood and blood
plasma. In the first case ashing must precede the determination
and evaporation be omitted. In the case of plasma the procedure
may be carried through in the same way as for serum potassium,
with heparin as the anticoagulant.
It is essential in serum potassium determination that hemolysis
be prevented and the cells centrifuged out as rapidly as possible
after the blood is drawn. The presence of hemolyzed red cells
invariably causes high results.
SUMMARY.

1. The method for determination of potassium in blood sera by

the indirect method is modified as given above, with an improve-
ment in the speed and accuracy.
2. Data are presented showing the limits of accuracy and the
extent of duplication of the method.
3. The potassium values obtained in a few normal and patho-
logical conditions are given.
The author acknowledges the assistance afforded him by his
colleague, Dr. A. G. Young of this laboratory, in providing tech-
nical assistance.
BIBLIOGRAPHY.

1. Kramer, B., andTisdal1, F. F., J. Biol. Chem., 46,339 (1921).

2. Clausen, S. W., J. Biol. Chem., 36,479 (1918).
3. Briggs, A. P., J. Biol. Chem., 67,351 (1923).
4. Ilosvay, N. I., cited by Treadwell, F. P., Analytical chemistry, trans-
lated by Hall, W. T., New York, 6th edition, 306 (1924).
5. Personal communication, quoted by permission of Dr. Bernhard
Domogalla.
6. Taylor, F. H. L., unpublished data, Dissertations, University of Wis-
consin (1926).