U.S. Army Medical Department, Office of Medical History
Skip Navigation, go to content

HISTORY OF THE OFFICE OF MEDICAL HISTORY

AMEDD BIOGRAPHIES

AMEDD CORPS HISTORY

BOOKS AND DOCUMENTS

HISTORICAL ART WORK & IMAGES

MEDICAL MEMOIRS

AMEDD MEDAL OF HONOR RECIPIENTS External Link, Opens in New Window

ORGANIZATIONAL HISTORIES

THE SURGEONS GENERAL

ANNUAL REPORTS OF THE SURGEON GENERAL

AMEDD UNIT PATCHES AND LINEAGE

THE AMEDD HISTORIAN NEWSLETTER

Chapter 10

Chapter 10

In Vitro Studies of Whole Blood During Shipment and Storage

            Major Edward C. Knoblock, MSC, USA
            First Lieutenant John M. Olney, MC, USAR
            Lieutenant Colonel William H. Crosby, MC, USA
            Captain John M. Howard, MC, USAR
            First Lieutenant Harry R. Pilhorn, MSC, USAR
            First Lieutenant George R. Haynes, MSC, USAR

This study was designed for the following purposes:

1. To describe the changes of plasma hemoglobin, plasma potassium, whole blood glucose, and red cell osmotic fragility in bank blood collected and transported through the regular blood program channels.

2. To compare the in vitro quality of whole blood at varying periods of time following collection.

3. To compare whole blood collected, shipped and stored in bottles, and that in plastic bags.

Materials and Methods

This study was undertaken as a cooperative effort between personnel of the Department of Chemistry, 406th Medical General Laboratory, in Japan, and the Surgical Research Team in Korea.

Blood samples for examination of bottled blood were selected at random from stocks shipped from the United States. Collection of blood in plastic bags was done at the Blood Processing Laboratory, Travis Air Force Base, California, and these bags were shipped simultaneously with the regular quotas of bottled blood. For control studies of the effect of handling of blood for long shipments, collections of blood were made by the Blood Bank Laboratory Detachment, 406th Medical General Laboratory. All bloods were stored at a temperature of 2 to 6 C until examined and were transported through normal blood shipment channels.

Comparative studies of plasma hemoglobin, potassium and red cell osmotic fragility are reported herein. A series of whole blood glucose levels on preliminary shipments of blood is not recorded since it was found during the study that anticoagulant solutions of both NIH formula A and B were used in making collections. It was not possible to determine the ACD solution used in any instance. Only in


142

the study of bottled blood and plastic bags, received as one shipment, are the glucose values reported.

Sampling

Bottles of blood were selected at random from stocks received and maintained by the local blood bank. Blood selected was that which would normally have been dispensed to the surgeon. The bottles were mixed by rolling from side to side for a period of 5 minutes. Bags were mixed by inversion for a similar period. Shaking was avoided. The selected unit of blood was sampled using a dry syringe and size 17 needle. Plasma samples for the studies were separated from the cells as soon as possible after collection.

Plasma potassium was determined by flame photometry using lithium as an internal standard. For convenience, the analyses done in Korea were on plasma samples stored under refrigeration for 3 or 4 weeks after they were drawn from the bottle of blood. Control studies demonstrated no change in potassium concentration during storage.

Plasma hemoglobin was determined by the method of Bing and Baker as modified at the 406th Medical General Laboratory.

Into a suitable clorimeter cuvette 0.6 ml. of 2 per cent benzidine base in glacial acetic acid, 0.2 ml. of suitably diluted plasma sample, and 0.2 ml. of 0.6 per cent hydrogen peroxide were placed in sequence named. After 1 hour 6 ml. of 20 per cent acetic acid was added and the transmission compared at wave length 640 m to standards of 5.0, 7.5 and 10.0 mg. per 100 ml. hemoglobin which were prepared by diluting whole blood. The unknown was diluted with normal saline to approximate the range of the standards. A Coleman Junior Spectrophotometer was used for these studies.

Red cell osmotic fragility was determined in phosphate-buffered saline solutions as described by Parpart et al.1 Reagents for this examination were prepared as a single lot at the 406th Medical General Laboratory and used by both laboratories. A special shipment of Cutter and Fenwal plastic bags required preparation of a second lot of reagents.

Whole blood hematocrit was used to evaluate mixing of the blood samples. In any instances where the hematocrit varied by more than 5 per cent from day to day samplings, the determination was invalidated. Culture for bacteria was used to check the continued sterility of samples. Any bags or bottles which were found to be contaminated were eliminated from the study. One bag and two bottles were eliminated for this reason.


143

Results

The study was divided into three phases which are reported as follows:

1. Comparison in Japan and Korea of bank blood collected in the United States to evaluate the effect of shipping and storage.

2. Comparison of different types of plastic bags used for collection, transportation and storage of whole blood.

3. Comparison of whole blood collected and shipped in plastic bags and blood which was similarly processed in standard blood bank bottles.

Studies of Blood from the United States. This group consists of blood collected in the United States in standard bottles using ACD solutions. Samples were obtained from regular shipments to the Far East Command.

Plasma potassium and hemoglobin values are presented in Table 1. There is a regular, almost linear, rise of plasma potassium with time which may be extrapolated to 4.0 mEq./liter at time of drawing. The rise continues to approximately 20.0 mEq./liter on the twentieth day and thereafter rises more slowly to reach 24.0 mEq./liter on the thirtieth day. Individual specimens showed a relatively small scatter about the mean and the standard deviation appeared independent of the age of the blood.

Plasma hemoglobin levels showed a much wider scatter between individual observations; there was an average rise from 10 to 15 mg. per 100 ml. on the fifth day to 100 mg. on the twentieth day. When an attempt was made to compare plasma hemoglobin and potassium values, only an approximate correlation with occasional marked difference was noted on individual specimens. The two often appeared to vary independently.

Osmotic fragility of red cells showed a considerable variation between samples and could not be accurately correlated with the age of the blood. The mean of the examination of 10 units of blood in each sampling at the saline concentration of 0.6 per cent as suggested by Sack et al.2 indicates increased fragility of red cells with time in the Tokyo-examined blood and also in that blood examined in Korea (Table 1).

Comparison of Different Types of Plastic Bags. Three separate shipments of plastic bags were received and examined. These included two types of plastic bags and different ACD concentrations as follows:
 

Identification

Description

Anticoagulant Solution

Group 1

Cutter Bag

120 ml. ACD, NIH Formula "B"

Group 2

Fenwal Bag

75 ml. ACD, NIH Formula "A"

Group 3

Fenwal Bag

120 ml. ACD, NIH Formula "B"


144

Table 1. Plasma Potassium and Hemoglobin and Osmotic Fragility of Whole Blood Shipped From the United States in Bottles
 

 

Plasma K (mEq./1.)

Plasma Hb. (mg. per 100 ml.)

Osmotic Fragility % Hemolysis in 0.6% Saline

Age (Days)

No. of Samples

Mean

Std. Deviation

Mean

Std. Deviation

Studied in Tokyo

5

10

9.1

1.5

16

7

8

6

10

10.9

2.5

17

6

3

8

10

9.5

1.8

38

 

4

10

10

11.9

3.5

30

20

10

12

10

13.0

1.3

23

13

11

14

10

15.5

3.0

62

39

11

16

10

19.4

6.6

134

104

10

18

10

18.3

2.7

83

58

17

20

10

19.4

4.0

143

67

23

22

10

18.3

3.5

79

74

28

24

10

20.6

 

79

 

26

26

8

19.2

2.3

79

69

40

28

8

19.7

 

108

 

23

30

10

23.4

3.4

194

70

38

Studied in Korea

*7 5

9.9

 

5

52

 

3

9 2

11.2

 

2

76

   

11 9

14.0

2.0

13

61

60

7

13 10

14.5

2.3

13

68

21

8

15 10

16.5

1.4

10

66

50

6

17 11

16.6

2.9

13

43

 

8

19 9

18.2

1.6

15

87

 

17

21 9

19.0

2.3

11

113

 

7

25 3

20.3

 

11

142

 

12

30

   

10

209

   

*Blood collected in Japan.

Results of these examinations are reported in Table 2. On gross examination small clots were noted in three specimens of Group 3 (two of them 6 days old and one 10 days old) and small coagulated masses, thought to be fibrin, which disappeared on mixing were seen in most specimens of Group 3 examined in Japan. Examinations in Korea did not note any such phenomena.


145

Table 2. Blood Stored in Plastic Bags
 

 

Plasma K

Plasma Hb.

Blood Glucose

Osmotic Fragility % Hemolysis in 0.6% NaC1

Group

Age (Days)

No. Samples

(mEq./1.)

mg./100 ml.

mg./100 ml.

A. Studied in Tokyo

1

6

2

9.4

10

316

8

1

8

2

11.2

22

321

6

1

10

2

12.2

22

287

2

1

12

2

14.0

20

333

2

1

14

2

14.4

32

260

4

1

16

2

18.4

94

285

2

2

6

2

14.4

96

390

17

2

8

2

14.5

46

372

7

2

10

2

23.5

64

380

8

2

12

2

17.3

57

332

17

2

14

2

18.7

86

327

9

2

16

2

24.5

97

313

18

3

6

2

14.4

5

605

 

3

8

2

19.2

16

558

 

3

10

2

23.4

40

543

 

3

12

2

23.4

39

572

 

3

14

2

27.0

43

553

 

3

16

2

29.3

56

456

 

B. Studied in Korea

1

4

4

8.8

15

346

2

1

11

4

15.2

15

358

4

1

20

4

21.6

35

310

4

2

5

4

15.0

18

332

8

2

11

4

20.9

52

248

 

2

21

4

31.5

145

309

17

3

10

12

21.9

12

615

 

Estimated Standard Deviation 10%

50%

   

In Group 1 the plasma hemoglobin rose from approximately 9.0 mEq./l. on the fifth day to 21.5 mEq./1. on the twentieth day, which is similar to the rise noted in bottled blood. Groups 2 and 3 show a more rapid rise in plasma potassium. Group 2 rose from 12.0 mEq./1. on the fifth day to 24.0 mEq./1. on the sixteenth day and Group 3 rose from 15.0 mEq./1. on the sixth day to 30.0 mEq./1. on the


146

sixteenth day. The control bottled blood rose from 9.5 mEq./l. on the sixth day to 16.5 mEq./l. on the sixteenth day.

The relationship of plasma hemoglobin to time was not as sharply defined as plasma potassium. The increase in plasma hemoglobin was generally slower in the plastic bags than in the bottled blood.

Red cell osmotic fragility examinations are probably not adequately sampled to demonstrate a mean difference. Before the fifteenth day all groups are similar to bottled blood and after this time Group 1 appears lower. Group 2 and Group 3 were not examined after 16 days.

Comparison of Whole Blood Collected and Shipped in Plastic Bags and Blood Similarly Processed in Standard Blood Bank Bottles. A shipment of 24 Fenwal plastic bags and 24 Baxter bottles of group O whole blood, preserved with 120 ml. NIH formula 1 ACD solution, was received for examination. Each of these units of blood was stored at normal blood-bank temperatures and sampled on alternate days. The bloods were collected in the Zone of Interior and handled by the normal channels for blood shipment. The plastic bags were 5 days old when received and the bottles 6 days.

Plasma potassium (Fig. 1) was higher in the bags on initial examination and increased more rapidly for the entire period of examination when compared to the bottled whole blood. The rate of change in both bags and bottles was a steady one with the bags showing 13.4 mEq./l. plasma potassium on the fifth day and 36.9 mEq./1. on the thirtieth day. The bottled whole blood showed 11.2 mEq./1. on the sixth day and 24.4 mEq./1. on the thirty-first day of examination. Standard deviation of each lot was relatively constant for the entire period of examination, with more than 90 per cent of each sampling falling within one standard deviation from the mean.

Plasma hemoglobin values (Fig. 2) were slightly higher in the bottled blood until 16 days, after which time the blood in the plastic bags showed accelerated hemolysis. The blood in plastic bags showed a gradual increase from 5.3 to 29.2 mg. per 100 ml. hemoglobin between the fifth and fifteenth day and then climbed rapidly to 231 mg. on the thirtieth day. The bottled blood showed 10.7 mg. per 100 ml. hemoglobin on the sixth day and 32.6 mg. on the sixteenth day. Rate of increase was not as rapid in the bottles as in the bags after 16 days and at 31 days the plasma hemoglobin of the bottled blood was 112 mg. per 100 ml. Standard deviation in both samplings increased with age of blood. Most of the values fell within one standard deviation from the mean.

Osmotic fragility (Fig. 1) of the red cells in the plastic bags was, as a rule, lower than that of the bottled blood. After 15 days the blood in the bags showed decidedly less hemolysis in 0.6 per cent saline than that preserved in the bottles.


147

FIGURE 1.

Whole blood glucose (Fig. 2) in the bottles was lower at all times than in the bags. Apparently, a different lot of ACD solution had been used in the two types of containers. The glucose in the bottled blood decreased at a steady rate through 22 days and then its consumption was apparently accelerated for some reason. This did not


148

FIGURE 2.

occur in the bag blood, which continued glycolysis at approximately the same rate for the entire period of testing.

Summary and Conclusions

A regular, almost linear rise in mean plasma potassium was noted with time. The rise continued to approximately 20.0 mEq./l. on the twentieth day and thereafter rose more gradually to reach 24.0 mEq./l. on the thirtieth day. The plasma potassium could be


149

extrapolated to 4.0 mEq./l. at the time of bleeding. The individual specimens showed a relatively small scatter around the mean with the standard deviation apparently independent of the age of the blood.

Plasma hemoglobin levels in bottled blood showed a much wider scatter between individual observations with the standard deviation apparently more dependent on individual sampling than on time of storage. A rise in mean plasma hemoglobin was noted from 10 to 15 mg. per 100 ml. on the fifth day to 100 mg. on the twentieth day. Only approximate correlation was noted between the rise of potassium and hemoglobin in the plasma. These two values often appeared to vary independently.

In bottled blood (ACD solution, NIH formula A) the whole blood glucose fell at a steady rate from a mean of 312 mg. per 100 ml. at 6 days to a mean of 248 mg. at 22 days. Thereafter the rate appeared to accelerate slightly until at 31 days the mean glucose level was 174 mg. per 100 ml. The blood in bags showed a similar rate of decrease, but the slope at 25 days had a tendency to level off.

Comparison of bottled whole blood and that collected in plastic bags revealed the following information:

1. The rate of diffusion of potassium was more rapid in the whole blood collected and shipped in plastic bags for the entire period of comparison.

2. The mean osmotic fragility of red cells, using 0.6 per cent saline as a point for comparison, was more stable in the whole blood contained in plastic bags after 12 days. Up to this point the results were similar.

3. Plasma hemoglobin values in bags and bottles were comparable until 18 days of storage. After this time the plasma hemoglobin in the plastic bags increased more rapidly than in the bottles.

References

1. Parpart, A. K., Lorenz, P. B., Parpart, E. R., Gregg, J. R., and Chase, A. M.: The Osmotic Resistance (Fragility) of Human Red Cells. J. Clin. Invest. 26: 636-640, 1947.

2. Sack, T., Gibson, J. G., and Buckley, E. S.: The Preservation of Whole ACD Blood Collected, Stored, and Transfused in Plastic Equipment. Surg., Gynec. & Obst. 95: 113-119, 1952.

RETURN TO TABLE OF CONTENTS