STUDIES IN BLOOD PRESERVATION The Serum Potassium of Cadaver Blood JOHN SCUDDER, M.D., Med. Sc.D., F.A.C.S., DOROTHY R. CORCORAN, M.A., and CHARLES R. DREW, M.D., C.M., New York, New York N observed increase in plasma potassium of a patient dying from an automobile accident (9) suggested a further in- vestigation of this base in cadaver blood, especially as this source has been used extensively for transfusions (12). The concentration of potassium within the intact red blood cells is twenty times that of the plasma. In this respect, the erythrocyte resembles the marine alga, valonia macro- physa. In the latter, Osterhout (5) explains the higher internal concentration of potassium as due not to the formation of insoluble com- pounds, but due chiefly to diffusion constants and concentration gradients, the latter de- pending on partition coefficients (7, 8). Osterhout’s (6) quantitative studies on cell sap of valonia have revealed a loss of po- tassium to be associated with injury. An- other factor which causes these cells to lose potassium is an increase in ammonia. Jacques and Osterbout have shown that if the concentration of ammonia in sea water were raised by 0.coor molar, there occurs a rapid exit of potassium from these cells. Methods. Under sterile precautions the blood was collected from the heart on a num- ber of unselected cases; one portion was cul- tured, and the remainder was introduced into a Sanfort-Magath hematocrit tube and cen- trifuged for one hour.- The average time of separation of serum from the cells was 35 hours after death. Potassium was determined by the argenticobaltinitrite method (10, 11), the final color being read on an Evelyn photo- electric colorimeter (2). Results. The average concentration of the serum potassium in these 27 cases was 101 milligrams per cent. These results contrast From the Department of Surgical Pathology, College of Physi- cians and Surgeons, Columbia University. A study made possible by a grant from the Blood Transfusion Betterment Association, with our average of 17.2 milligrams per cent in the plasma of normal venous blood and the average of 29.7 milligrams per cent in the plasma of cardiac blood taken at death (9) (Table II). HIGH SERUM VALUES IN CADAVER BLOOD This high serum potassium approximately 3.5 times the concentration of that found at death and nearly 6 times greater than normal illustrates the rapid diffusion of potassium in ‘cadaver blood. It takes refrigerated fresh 48 blood 110 hours to reach approximately this - same concentration. Factors affecting this rapid diffusion are many. Among them tem- perature is one, for preserved blood is usually placed in the icebox immediately, whereas many hours may elapse between death and storage of the cadaver. A second factor may be the ammonia concentration. There has been much difficulty in determin- ing the free ammonia in circulating blood. Folin and Dennis were the first to find values under o.1o milligram ammonia-N per 100 milliliters. Subsequently a value of 30 gamma (0.03 mg.) per 100 milliliters was considered to represent the free afamonia-N in circulating human blood. Conway with his isothermal distillation method has. established the true value to be “less than one-tenth this amount, or less than one part in thirty million.” Our interest in ammonia resulted from our curiosity about the high serum potassium val- ues in cadaver blood. With the cobaltinitrite method it has long been known that if am- monia is present, there is a quantitative pre- cipitation of ammonium cobaltinitrite along with the potassium salt. 3 NH, + Co(NO2). aaneeal (NH,)sCo(NOz). 3 K + Co(NOz)¢ _ K;Co(NOs). Subsequent analysis for ammonia on several cadaver bloods gave the highest value to be SCUDDER, CORCORAN, DREW: STUDIES IN BLOOD PRESERVATION TABLE I.—SERUM POTASSIUM OF CADAVER 49 TABLE Il.—PLASMA POTASSIUM OF HUMAN CARDIAC BLOOD OBTAINED AT AUTOPSY BLOOD* Hos- Serum Plasma potassium expressed as pital | Initials potas- a sex, and Principal autopsy findings mai Number Milligrams Millimols of age grams 3 ane les Source of sample per cent per liter death per cent analyzed 4998] J.M., | Rheumatic endocarditis . Aver- Aver- 7338 M.,27 | - 547 age Range age Range 531679| T.G., | Carcinoma of stomach; duodenojejunos- 50.2 73 | Venous blood from 17.2 |13.5- 21.5] 4.4] 3.5- 5.5 3-31-38] M., 45 omy normal young adults 37890 | J.G., Tuberculous leptomeningitis 60.9 13 | Cardiac blood remov- | 29.7 | 24.0- 38.0/ 7.6 | 6.1- 9.7 5-10-38) F., 3x ed at death . 37862 | C.V., Glioblastoma of cerebral hemisphere 64.5 27 Cardiac blood remov- | 101.0 | 54.7-180.0 25.8 | 14.0~46.0 8-18-38] M., 57 ed at autopsy MH, ryperplasi: id with hyper id- *Centrifuged at 2,000 r.'p, m. for 1 hour after which the plasma or serum ares F., 50 Hype lasia of thyroid with b thyroid-| 70.0 was immediately separated from the cells. The average time of separating the serum from the cells after death L. i j iac-| was 31.5 hours. ss B 56 cer eeet tanh formation of b fart, cardiac-| 70.4 The gure for each determination represents the mean of 2 aliquot samples. 11647 | H.LaC. | Multiple fractures of skull : 4-6-38| M., 60 Pi 72-0 7 ADDING 0 37736 | C.O°K., | Abscesses of cerebellam a TABLE UI.—THE EFFECT OF ADDING AMMONIA 4-25-38] F., 35 TO BLOOD ON POTASSIUM DIFFUSION o 487166/ A.R., | Cardiac hypertrophy and dilatation, clin- | 76.6 TEMPERATURE 38 C. 7-22-38] M., 53 | ical diagnosis: hypertension, uremia ' 22385] L.H., | Chronic myeloid leucemia . Potassium as milligrams per cent in sre 38 9 4I j 77-4 ee plasma of citrated blood ein 289643| M.F., Carcinoma of female gland 77.7 Date hours from 4-19-38) F., 35 with multiple metastases to lymph nodes phlebotomy iated | Difference - - | Control oe between 548449} L.B., Placenta previa, cesarean section 80.8 specimens 4-27-38] F., 27 —18—; 20.9* 345832|G.H., | Carcinoma of stomach, partial gastrec- 87.2 sre 39 a 8 4-11-38] M., 66 tomy . 2 19.6 30.6 IL.o 3652| R.R., Subacute glomerul hritis, ia, . . . . 3-2-38| M., 62 | “clinical dingnosia: Brora mnie, | 97.0 4 nee “8° a 6 . . II. 552683) B.T., Acute gangrenous appendicitis with peri- 98.0 “s cad 4 6-21-38] M., 56 tonitis 3-19-39 8 alg 33-2 12.3 gse7asg ee Carcinoma of sigmoid, resection of colon 98.3 15 23.0 40.0 17.0 8 +2 . 6. 451845| HLR., Lymphosarcoma of lymph nodes, second- 98.9 I 27 43.9 16.7 4-30-38) F., 48 ary phosarcoma of dura mater, multi- 23 30.9 52.8 22.9 ple abscesses, inectomy, resection of or 3-20-39 3r 40.4 55-7 15.3 250743| S.F., Arteriolar nephrosclerosis; clinical diag- 110.0 39 79.9 98.1 18.2 4-27-38] F., 64 nosis: uremia 3-21-39 Es 121.0 120.0 1.0 284144! W.EL., i iti. p-tead M., 13 Rheumatic endocarditis 718.0 *Plasma potassium of undiluted blood. Each Tepresents the mean S of two aliquots. Control: 225 c. em. of blood 25 c. cm. of 2.5 per cent 3762, 5.5 | Meningi If groovi ion: sodium citrate. . 59-38 F., 52 bilateral cetvical syznbathertore ee sid . oniated blood: 250 c. cm. of blood, 25 c. cm. of 2.5 per cent sodium citrate, pnd gM. Subacute bacterial eudocendttig due to 118.6 “31-3 +2 40 gram positive coccus, splenectomy eae eayeqs 543474 ae Carcinoma of colon, colostomy; resection 128.0 1.3 milligr: ams per 100 ters. . We con 474-38) M.,'67_| “of carcinoma of eclon cluded, therefore, that the apparent high po- 852258] V-E., | Carcinoma of stomach r4r.0 tassium values were not due to ammonia but 3 * . ° . . ; — were genuine. This concentration of ammonia, a UR. i locarditi . > : 37-38] Fy gx_| Shenmatic endocarditis “ee however, is far in excess of that which Jacques 549302] K.S., Acute splenic tumor, clinical cause of 66, i 225-38] Mobs_| Agute,splenic tumor, clinical cause 166.0 and Osterhout have shown causes potassium Sa6r47 KB, Venous angioma, temporal lobe 175.0 to leave valonia. we 48-38) F., 63 - . ; To ascertain whether the addition of am- 545415] R.M., | Carcinoma of kidney, pneumonia 180.0 : oe . . * 4717-381 M,, 58 monia causes a similar diffusion of potassium 50 SURGERY, GYNECOLOGY AND OBSTETRICS from red cells, sufficient ammonium chloride was added to one of two similar portions of citrated blood to bring its concentration to approximately o.o1 molar. These specimens were stored in identically shaped bottles in a water bath kept at 38 degrees centigrade throughout the experiment (Table ITI). The results indicate that with the addition of ammonium chloride, a prompt rise in the plasma potassium takes place and the rate of diffusion of potassium is more rapid in the ammonia flask than in the control during the first seventy-one hours of the experiment. SUMMARY The increase in serum potassium is more rapid in cadaver than in fresh blood stored in a refrigerator at 4 degrees centigrade. BIBLIOGRAPHY 1. Conway, E. J. Apparatus for the micro-determination of certain volatile substances. IV. The blood am- to. qi. I2. monia, with observations on normal human blood. Biochem. J., 1935, 29: 2755-2772. Evetyn, K. A. A stabilized photoelectric colorimeter with light filters. J. Biol. Chem., 1936, 115: 63-75. Fotty, O., and Dennis, W. Protein metabolism from the standpoint of blood and tissue analysis. J. Biol. Chem., 1912, 11: 161-167. Jacgurs, A. G., and OsrERHouT, W. J. V. The ac- cumulation of electrolytes. III. Behavior of sodium, potassium and ammonium in valonia. J. Gen. Physiol., 1930, 14: 301~324. . OstErHouT, W. J. V. Some aspects of selective ab- sorption. J. Gen. Physiol., 1922, 5: 225-230. . Idem. Some Fundamental Problems of Cellular Physi- ology. New Haven: Yale University Press, 1927. Idem. Permeability in large plant cells and in models. Ergebn. d. Physiol. u. exper. Pharmak., 1933, 35: 967~1021. Idem. The absorption of electrolytes in large plant cells. Botanical Review, 1936, 2: 283-315. . ScuDvER, J., SmirH, M.E.,and Drew, C. R. Potas- sium content of cardiac blood at death. Am. J. Physiol., 1939, 126: 337-340. TruszKowski, R., and ZWEMER, R. L. Corticoadrenal insufficiency and potassium metabolism. Biochem. J., 1936, 30: 1345-1353. . . Idem. Determination of blood potassium. Biochem. J-1 1937, 31: 229-233. unin, S. S. Transfusion of stored cadaver blood. Lancet, 1937, 2: 361-366. .