THE ZINC CONTENT OF NORMAL HUMAN WHOLE BLOOD, PLASMA, LEUCOCYTES, AND ERYTHROCYTES*

BY BERT L. VALLEE AND JOHN G. GIBSON, 2ND

(From the Medical Clinic of the Peter Bent Brigham Hospital, the Department of Medicine, Harvard Medical School, Boston, and the Department of Physics,

Massachusetts Institute of Technology, Cambridge)

(Received for publication, July 11, 1948)

The literature contains few references to the quantitative determination of zinc in human blood. It is difficult to compare the scant data which are available because of the divergence of the methods used and of the units in which different authors chose to express their results (l-5).

It became apparent in the pursuit of our work on zinc in leucocytes and red blood cells, preliminary reports of which have appeared elsewhere (6-8), that the data available in the literature did not offer an adequate point of reference for our studies. A technique was developed (9) for the microanalysis of zinc in biologic materials. This method is sensitive for quantities as small as 1 y (microgram) and its limits of precision are de- fined by a standard deviation of flO.O per cent. This technique was employed throughout our work.

Arc spectrography’ of leucocytes and erythrocytes had demonstrated that, for equal numbers of cells, zinc was present in higher concentration in leucocytes than in erythrocytes. We therefore decided to examine whole blood, plasma, washed erythrocytes, and washed leucocytes sepa- rately for their zinc content.

Method

Venous blood samples were drawn into a 20 cc. zinc-free syringe (9) which had previously been wetted with heparin. Red and white blood cell counts were done in standard hemocytometers, in duplicate. Only counts agreeing within 5 per cent were accepted. Hematocrits were carried out in Wintrobe tubes, spinning 30 minutes at 3000 R.P.M. Hemo- globin was determined by the method of Evelyn (10). Differential counts of leucocytes were made from smears stained with Wright’s stain, 300 cells being counted.

A separation of red cells, white cells, and plasma was performed by the

* This study was supported by a grant-in-aid from the National Institute of Health.

1 Through the courtesy of Dean George R. Harrison and Mr. Rockwell Kent, 3rd, Massachusetts Institute of Technology.

445

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446 ZINC CONTENT OF BLOOD

flotation technique (11). All of the separated leucocytes were removed from the flotation tube and washed in a 0.02 per cent saponin solution in 0.85 per cent NaCl, until all entrained red cells had been removed, as evidenced by the absence of hemoglobin in the supernatant washing. Zinc analysis was carried out on the entire mass of washed white cells.

A portion of the red blood cells was transferred from the flotation tube to a centrifuge tube and washed in 0.85 N saline by centrifugation. The washing was discarded, and an aliquot of the packed cells was transferred to a 4 cc. hematocrit tube and made up to volume with 0.85 per cent NaCl solution. The quantity of cells was such that the final hematocrit was about 30. Two further washings were performed, in the 4 cc. tube, and from the third and final dilution a red blood count (in duplicate), hemato- crit reading, and hemoglobin determination were obtained. The duplicate red blood counts were required to check within ~5 per cent. These 4 cc. samples of red blood cell suspensions were the final aliquots used for analysis.

The measurement of the zinc content of whole blood, plasma, leucocyte, and erythrocyte samples was performed as previously described (9). Routinely 2 cc. of whole blood and 3 cc. of plasma were analyzed.

Calculation of Results-Zinc concentrations in whole blood and plasma were expressed in micrograms per cc. analyzed. Since volumetric measure- ment of white cells (by hematocrit) was impractical, due to the limited mass of white cells obtainable and the large variation in dimensions of the various classes of white cells, unit zinc concentrations for leucocytes were calculated in micrograms per million cells. Unit zinc concentrations for red cells also were calculated in micrograms per million cells for comparison with unit white cell zinc concentrations, and in micrograms per cc. of packed cells. Examples of calculations of results for leucocytes and erythrocytes follow.

The total number of cells in the sample of leucocytes was determined by multiplying the white cell count per c.mm. by the number of c.mm. of whole blood floated. For example, the white blood cell count = 1 X lo4 cells per c.mm. ; whole blood floated - 15 cc.; total cells in 15 cc. of whole blood = 1 X lo4 X 1 X lo3 X 1.5 X 10’ = 150 X lo6 cells; total Zn in sample = 3.0 y; Zn per 1 X lo6 cells = 2 X 10e2 y.

The total number of red cells in the sample was determined by multiply- ing the red count of the final dilution of cells by the final volume of that suspension. For example, the red blood cell count on final red cell dilution = 3.5 X lo6 per c.mm.; total cell suspension analyzed = 4.0 cc.; thus 3.5 X 10” X 1 X lo3 X 4 = 14,000 X lo6 cells; total Zn in sample = 17.5 y; Zn per 1 X 10” cells = 1.25 X 10m3 y.

The zinc in cc. of packed cells was calculated on the basis of micrograms

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B. L. VALLEE AND J. G. GIBSON, 2ND 447

of zinc per million cells, red blood cell count, and hematocrit of whole blood, as follows: the red blood cell count on whole blood (undiluted) = 5 X lo6 per c.mm.; hematocrit of whole blood = 45.0 per cent; thus (1.25 X lo-+ X 5 X lo6 X 1 X 103)/(4.5 X 10-l) = 13.9 y per ml. of packed cells.

As a check on the accuracy of the individual measurements of whole blood, and the three blood components analyzed, an internal check was calculated for each blood sample. The amount of zinc in 1 cc. of whole blood due to the zinc content of white cells, red cells, and of plasma was calculated as follows: (total Zn in all white cells analyzed)/(l5 cc.) = Zn in 1 cc. of whole blood due to leucocytes; ((Zn)/(million red blood cells)) X ((red blood cells)/( cc. of whole blood)) = Zn in 1 cc. of whole blood due to erythrocytes; Zn in 1 cc. of plasma X (1 - (hematocrit)/lOO) = Zn in 1 cc. of whole blood due to plasma; = total Zn in 1 cc. of whole blood.

The deviation of the total whole blood zinc so calculated and the directly determined zinc content of whole blood was taken to be a measure of the over-all accuracy of the hematological procedures and of the preliminary processing, including dry ashing, as well as the chemical analyses for zinc.

Material Studied-Thirty-one normal individuals, fifteen males and six- teen females, ranging in age from 18 to 45 years, were studied. Repeated determinations were carried out in six of these subjects, bringing the total of single analyses to thirty-eight. All blood samples were drawn in the morning with the subject fasting but not otherwise under basal conditions. Each sample was analyzed for all of the data described above under “Method.” In a few instances one of the components was lost in proc- essing.

In addition, five patients, with no apparent blood dyscrasias, on the medical wards of the Peter Bent Brigham Hospital, were studied. One had a peptic ulcer, one had recovered from bronchopneumonia, one had milkman’s disease, one was compensating from congestive heart failure, and one was suspected of having a brain tumor.

Fifteen consecutive blood samples taken at 1 to 4 day intervals from the same individual, Case 53, a 22 year-old normal male, were also analyzed.

Results

Unit zinc content, as described above, for whole blood and its three components for the group of normals and the patients is given in Table I, and for the series in one normal person, in Table II. Also shown in Tables I to IV are the means and standard deviations (s.D;) for all unit zinc concentrations.

The values for the amount of zinc contained in 1 cc. of whole blood due to plasma, leucocytes, and erythrocytes, calculated as described above, are shown for the normals and patients in Table III, and for Case 53 in

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448 ZINC CONTENT OF BLOOD

TABLE I Unit Values of Zinc Content of Whole Blood, Plasma, Leucocytes, and Erythrocytes

/

Unit content by ditbizone extraction Experiment No.

D&z 1 Whole blood , Plasma 1 Leucocytes 1 Erythrocytes

Normal males

23-l May 9 7.7 2.6 23-2 “ 12 9.2 2.2 29-l “ 16 7.6 1.2 34-l “ 26 10.5 4.3 36-l “ 28 7.5 3.2 73-l Sept. 11 13.7 11.4 80-l “ 29 5.9 2.0 80-2 Oct. 8 8.1 3.7 84-1 “ 2 6.3 1.9 85-1 (’ 2 7.0 3.0 88-l “ 6 9.5 4.1 88-2 “ 20 7.9 1.7 89-l “ 6 8.3 3.2 90-l “ 7 8.6 4.1 99-l “ 16 9.1 5.5 108-l “ 24 9.2 3.8 109-l “ 24 14.8 9.1 122-1 Nov. 19 9.3 3.7

Mean ...... S.D .........

1947

24-l 25-2 25-3 26-l 28-l 28-2 31-1 76-l 77-1 82-l 86-l 86-2 86-3 87-l 93-l

-~

May 12 June 26 Sept. 26 May 14

“ 14 Oct. 3 May 19 Sept. 18

“ 18 Oct. 1

‘I 3 “ 9

Nov. 5 Oct. 3

“ 9

-

-- -

T-

y per cc.

8.9 f2.0

y per cc.

3.9 12.5

Normal females

9.1 8.5 7.9 8.5 9.1 6.7 8.7 5.2 7.6

8.1 13.2

11.5

3.0 3.4 2.2 1.6 2.0 1.3 2.0 1.5 1.2 4.4 4.8 3.2

10.1 4.5 4.6

-

--

-

rX10-2 m&g 106

1.2 4.2 1.6 2.9 4.6 2.8 2.2 6.5 2.0 1.8 1.9 2.1 4.7 2.3 1.4 5.5 2.3

2.9 f1.5

-

--

-

YXIO- per 1 x 100

cdl2

1.21 1.35 1.25 1.41 1.20 1.20 0.97 1.20 1.30 1.15 1.71 1.19 1.44 1.20 1.28 1.59 1.83 1.29

1.31 14.2 ho.25 12.3

Zmy per cc. W$$Cd

13.2 14.7 16.4 15.7 12.6 11.9

9.1 12.3 12.7 12.2 17.0 15.2 16.6 14.1 13.6 17.1 18.3 18.4

4.8 3.8 2.6 3.5 2.9 6.2 4.2

1.7 3.6 4.4 2.1 3.1 2.9 3.1

1.26 13.8 1.32 13.6 1.36 15.0 1.06 11.3 1.15 13.5 1.30 13.6 1.27 13.8 1.09 16.1 1.21 12.5 1.60 16.7 1.63 17.1 1.38 13.9 1.54 16.5 1.42 12.8 1.54 17.0

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B. L. VALLEE AND J. cf. GIBSON, %D

TABLE I-Concluded

449

Experiment No. Date Unit content by dithizone extraction

Wholeblood [ Plasma 1 Leucocytes 1 Erythrocytes

100-l Oct. 17 101-l ‘I 17 115-1 Nov. 4 116-1 “ 5 123-1 “ 19

I 1947

Mea*. ’ ...... S.D ..........

Normal females-continued

y per CC. y per cc.

7.2 2.3 8.6 3.0 9.4 4.2

10.4 4.1 8.4 1.2

8.6 3.1 Al.8 f2.0

y x 10-p P”‘c’elx 106

3.5 4.2 1.9 4.6

-

-

3.5 f1.2

Y x 10-1 P”‘c’cls 10s

1.24 1.32 1.17 1.85 1.52

1.37 fO.195

_.

-

Zn y per CG. packed red

C&S*

11.9 14.2 12.9 18.4 15.5

14.5 Al.9

Total normals

Mean.. .I S.D..........'

___--

8.8 f2.0

-- - 30-l M. 48-l “ 40-l F. 50-l “ 61-1 “

Mean. . . . . . _-

3.0 / 3.2 1.34 14.4 f1.6 / f1.3 f0.20 f2.7

Patients with no blood dyscrasias

May 16 8.6 2.7 6.5 1.16 June 17 8.1 4.4 4.3 1.28

lL 4 8.5 4.2 2.0 1.43 July 21 12.3 8.7 2.4 1.33 Aug. 14 9.6 8.2 1.6 1.41

9.0 5.6 3.2 1.32

* Corrected for mean corpuscular volume.

13.8 13.3 14.9 14.6 16.0

14.5

Table IV. The total of the three values constitutes the “reconstructed” total blood zinc content.

DISCUSSION

In the evaluation of the data presented above, the limitations of the various hematological techniques employed must be taken into considera- tion. The errors inherent in the chemical analyses for zinc have been previously discussed (9).

Since Ostwald pipettes were used for the whole blood and plasma sam- ples, little additional error was introduced in the preparation of these samples for chemical analysis.

The limited quantity of leucocytes obtained from a routine sample of 15 cc. of blood precluded making a volumetric measurement on these

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450 ZINC CONTENT OF BLOOD

cells. It was not possible to make a final count on the leucocytes separated from heparinized blood because of clumping of cells, which persisted even after repeated washing with the saponin-saline solution. The “million cell unit” obviously reflects the errors inherent in leucocyte counts carried out in standard hemocytometers, although this error was reduced by dupli- cate counting. However, this unit allows conclusions as to the zinc con- tent of individual white cells, and is therefore of value in the study of

TABLE II Unit Values of Zinc Content of Whole Blood, Plasma, Leucocytes, and Erythrocytes in

Normal Individual (Case 63) Fifteen consecutive samples.

Experiment No. Date

1 2 3 4 5 6 7 9

10 11 12 13 14 17

July 31 Aug. 4

‘( 4 “ 5 “ 6 “ 7 “ 8 ‘( 12 “ 13 “ 14 “ 15 “ 18 “ 19 I’ 22

19 I ‘( 27

1947

Mean. ......... 9.1 3.9 2.5 1.47 15.5 S.D ............. f0.9 f1.3 fl.1 zkO.18 zkO.6

- Unit content by dithizone extraction

Plasma

y per cc. r per GG.

8.4 3.3 8.3 5.2 9.3 6.0 8.7 4.4 7.7 2.7

11.1 4.2 9.1 3.6 9.9 4.8 8.5 2.8 8.4 2.8

10.2 5.6 9.4 3.6 9.9 2.6 7.6 1.5 9.5 5.0

.eucocyte

-

s

Y x lo-, ‘C, 1 x 10

cells

1.2 2.2 1.7 2.0 4.2 2.0 5.1 2.0 2.0

1.9 1.5 3.5 1.9 3.7

Erythrocytes

TX/C- Zn y CC. ““‘;& 10% W$$Cd

1.48 15.7 1.37 14.5 1.63 17.9 1.43 15.0 1.20 11.7 1.77 18.7 1.51 15.9 1.40 15.1 1.69 16.4 1.66 16.5 1.66 17.2 1.14 12.5 1.44 16.5 1.19 12.5 1.59 17.1

* Corrected for mean corpuscular volume.

clinical conditions affecting leucopoiesis, and for comparison with cor- responding values for erythrocytes. The development of better techniques for handling leucocytes is now in progress.

The use of the million cell unit for erythrocytes also involves the same sources of error in about the same degree. However, the calculation of unit zinc content of red cells on the basis of hematocrit and red count permits comparison of unit zinc concentrations in anemias and other disturbances of erythropoiesis with the normal.

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B. L. VALLEE AND J. G. QIBSON, 2ND 451

Inspection of the data in Tables I and II shows that thevaluesfor whole blood, plasma, leucocytes, and erythrocytes in males, females, and in the totals vary over a fairly wide range. The measurements performed on one individual (Case 53) are of the same order of magnitude as found in the series of normals, though both their range and standard deviation are much narrower. They occupy the central region of a hypothetical distribution curve plotted for the data obtained from the series.

The variations and standard deviations of the white blood cell measure- ments are greater than those observed in the other categories. This could be predicted from the technical limitations imposed by the processing of leucocytes, as described above. Furthermore, the total zinc content of a leucocyte sample is of necessity small, because of the small number of white cells in circulation.

The larger aliquots of whole blood, plasma, and red cells obtainable for routine analysis contain about 5 times as much zinc as is contained in all the white cells from 15 cc. of whole blood. Since the percentage error of zinc extraction tends to be constant, the absolute error will of necessity be greater for white cells than in the case of whole blood, plasma, or red cells.

The distribution curves shown in Fig. 1 graphically express these facts, and are plotted from data obtained in normals given in Table III. The incidence is plotted for males, females, and total cases. Fig. 1, A is the curve for the directly measured whole blood zinc content. The curve is symmetrical and the mode closely approximates the arithmetic mean.

The distribution curves for plasma, Fig. 1, B, for leucocytes, Fig. 1, C, and for erythrocytes, Fig. 1, D, are plotted, not from the directly meas- ured unit values, but from the “reconstructed” value of zinc in 1 cc. of whole blood due to the particular component given in Table III. It was felt that the latter values more closely represented a fractional analysis of blood than did the direct determinations.

The curve for plasma, though nearly symmetrical, appears to be slightly skewed to the left. It will be noted that three of the cases fall well to the positive side of the mean. There was a reasonable probability that these values may have been high due to accidental contamination with zinc but, since the evidence is not conclusive, they are included in the series. The inclusion of these three cases raises the mean. If these are omitted, the mean falls to 1.53 y per ml., whereas the mode is approximated at 1.25. Thus biological significance cannot be attributed to the skewness of the curve.

The curve for leucocytes is grouped around a mean value of 0.21 y per ml., with a mode of 0.170. Here again the mean is slightly higher due to the inclusion of two cases with values much greater than the mean value plus twice the standard deviation. It should be stated that the absolute quan-

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452 ZINC CONTENT OF BLO.OD

tities of zinc measured in the samples are extremely small, so that losses in processing, which would not show up in samples of whole blood or of the other components, could be sufficient to account for the predominantly low values found in the majority of cases.

WHOLE BLOOD 201

a w tici 5.

3 z

o- 4 5 6 7 6 9lO II 12131415;r6

mictoqramr pet I ml.

LEUCOCYTES

0 A5 .I .I5 .2 925 .3 .35 .4 .45 .5 55 micrograms per ml. of wholo blood

due to lsucocyter

PLASMA

012345678 microqroms oer ml. of whole blood

due to plosmo

ERYTHROCYTES

207

15-

IO-

5

O- -r

34 5 6t i9lb

micrograms per ml. of whole blood

due to erythrocytes

FIG. 1. Distribution curves of zinc content of whole blood, plasma, leucocytes, and erythrocytes in normals.

The distribution curve for red cells, Fig. 1, D, is symmetrical and shows little skewness.

It is apparent that no significant difference exists between sexes in this series.

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B. L. VALLEE AND J. G. QIBSON, SD 453

TABLE III Zinc Content of 1 Cc. of Normal Whole Blood Calcutated from Unit Values of Plasma,

Leucocytes, and Erythrocytes All values in micrograms.

Normal males

23-l 1.4 23-2 1.2 29-l 0.7 34-l 2.4 36-l 1.8 73-l 6.2 80-l 1.2 80-2 2.3 84-l 1.0 85-l 1.5 88-l 2.0 88-2 0.8 89-l 1.6 90-l 2.3 99-l 3.1 108-l 2.1 109-l 4.9 122-1 1.4

Mean.... S.D... . . .

--

-

-

-

2.10 0.20 6.4 8.8 8.9 zkl.40 f0.095 11.25 zt2.03 f2.0

24-l 25-2 25-3 26-l 28-l 28-2 31-1 76-l 77-l 82-l 86-l 86-2 86-3 87-1 93-l

1.8 0.32 5.6 7.7 1.8 0.32 6.0 8.1 1.2 0.18 6.6 8.0 0.9 0.24 4.7 5.8 1.1 0.19 6.2 7.5 0.6 0.47 6.0 7.1 1.1 0.23 6.0 7.3 1.2 0.13 6.4 7.7 0.7 0.13 5.7 6.5 2.5 0.22 7.4 10.1 3.1 0.22 6.1 9.4 1.9 0.14 5.6 7.6 6.0 0.17 6.8 13.0 2.5 0.14 5.3 7.9 2.5 0.26 7.8 10.6

- 0.08 0.33 0.11 0.20 0.29 0.22 0.08 0.20 0.15 0.14 0.17 0.19 0.25 0.25 0.12 0.47 0.15

6.2 6.8 5.8 6.9 5.6 5.5 4.0 4.6 5.7 5.9 8.9 7.5 7.6 6.4 5.9 7.7 8.4 6.1

7.7 8.4 6.6 9.5 7.7

12.0 5.3 7.1 6.9 7.5

11.1 8.5 9.4 9.0 9.1

10.3 13.5

Normal females

7.7 9.2 7.6

10.5 7.5

13.7 5.9 8.1 6.3 7.0 9.5 7.9 8.3 8.6 9.1 9.2

14.8 9.3

9.1 8.5 7.9 8.5 9.1 6.7 8.7 5.2 7.6

8.1 13.2

11.5

--

-

-

-

0 -8.7

-13.2 -9.5 +2.7

-12.4 -10.3 -12.3 +9.5 +7.2

+16.9 +7.6

+13.3 +4.7

0 +12.0

-8.9

-0.1 ztlO.0

-15.4 -4.7 +1.3

-31.8 -17.6 +6.0

-16.1 +48.2 -14.5

-6.2 -1.5

-7.8

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454 7iRG CONTENT OF BLOOD

TABLE III-Concluded

100-l 1.4 0.17 4.9 101-l 1.7 0.30 5.6 115-1 2.4 0.13 5.7 116-l 2.5 0.30 7.9 123-1 0.6 7.0

Mean..... 1.80 0.22 6.2 8.2 8.6 -5.7 S.D... . . . . . AA.10 f0.09 hO.83 f1.65 f1.8 f16.4

6.5 7.6 8.2

10.7

7.2 8.6 9.4

10.4 8.4

-9.7 -11.7 -12.8

+2.9

Total normals

E?::::/ *:::: j *8:i 1 *:::5 / *% 1 *Ei 1 f124::

Patients with no blood dyscrasias

30-l M. 1.6 0.43 5.9 7.9 48-l “ 2.2 0.44 6.5 9.1 40-l F. 2.5 0.17 6.1 8.8 50-l “ 4.6 0.21 7.0 11.8 61-l “ 1.9 0.13 6.5 8.5

Mean.. . . . 2.56 0.28 6.4 9.3

8.6 -8.1 8.1 +12.4 8.5 +3.5

12.3 -4.1 9.6 -11.5

9.4 -1.7

Column 1, (Zn per cc. of plasma X 100) - (hematocrit of whole blood); Column 2, (total zinc in sample) f (cc. of whole blood from which leucocytes were obtained); Column3, Zn per million red cells X red cell count of whole blood X 1 X 103; Column 4, sum of Columns 1, 2, and 3; Column 5, zinc content of 1 cc. of whole blood di- rectly measured; Column 6, percentage difference between Columns 5 and 4.

The shapes of the curves for whole blood and the three components thereof indicate that the data presented are a measure of a biological norm, variable within physiological limits.

This fact is affirmed by the determinations carried out on the individual (Case 53) shown in Table IV. The mean values of the fractions and total blood zinc were close to those of the normal series but varied within much narrower limits, as indicated by the standard deviation.

Data on five patients who were suffering from illnesses other than blood dyscrasias are included, with their mean values. No standard deviation was derived because of the small number of patients involved. It will be noted that the unit zinc values lie within the range of values in the normal series.

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B. L. VALLEE AND J. G. GIBSON, 2ND 455

The percentage deviation in the reconstructed total ranged from -l-48.2 to -31.8, averaging -2.9, S.D. f14.3 for the normal series. The range for the five patients was +12.4 to -11.5, averaging -1.7. The range in Case 53 was from +20.0 to -17.0, averaging i-2.2, S.D. f11.0.

For statistical evaluation of the data, we have assumed that the direct determination on whole blood represents 100 per cent recovery of zinc,

TABLE IV Zinc Content of 1 Cc. of Whole Blood Calculated from Unit Values of Plasma, Leucocytes,

and Erythrocytes in Normal Individual Case 53, normal 22 year-old male; fifteen consecutive samples; all values in micro-

grams.

Experiment No.

1 1.9 2 2.7 3 3.3 4 2.4 5 1.6 6 2.4 7 2.0 9 2.7

10 1.6 11 1.5 12 3.1 13 1.9 14 1.4 17 0.9 19 2.7

Mean.. . . 2.14 0.20 6.9 9.2 9.1 +2.2 S.D... . . zkO.56 f0.06 zkO.88 Al.34 zko.92 fll .o

Plasma Leucocytes

- E

.-

:rytllrocytes -

.- Total Zn i;,t,c,c. of whole

-

- Per cent

deviations

0) (2) I (3) I (4) 6) 6)

0.10 0.19 0.15 0.19 0.29 0.19 0.35 0.25 0.16

6.7 7.0 7.8 6.7 5.2 8.1 7.0 6.6 7.3 7.4 7.6 5.8 7.2 5.5 7.8

8.7 9.9

11.2 9.3 7.1

10.7 9.4 9.5 9.1

8.4 8.3 9.3 8.7 7.7

11.1 9.1 9.9 8.5 8.4

10.2 9.4 9.9 7.6 9.5

+3.6 +17.0 +20.0 +6.9 -7.8 -3.5 +4.4 -4.1 +7.1

0.15 0.14 0.23 0.17 0.25

10.8 7.8 8.8 6.6

10.8

+5.9 -17.0 -11.1 -13.2 +13.7

Explanation of columns the same as in Table III.

The mean of the reconstructed total (Column 4, Table III) is slightly lower than the mean of the direct measurements.

The mean of the reconstructed whole blood zinc in Case 53 is slightly higher than the mean of the direct measurements.

The percentage deviation of Column 4 from Column 5, listed in Column 6 (Tables III and IV), brings these facts out more clearly. The mean percentage deviation of the series is -2.9 f 14.3, and for Case 53 (Table IV), the mean is +2.2 f 11.0. Therefore the reconstructed whole blood zinc level may be stated to fall within about f15 per cent of the zinc concentration found for whole blood by direct measurement.

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456 ZINC CONTENT OF BLOOD

It should be stated that the large range of percentage deviations actually represents minute differences in the absolute quantity of zinc present in the materials measured. Thus a loss or gain of 0.5 y from a total of 5 y is ~10 per cent, whereas the same quantity would constitute only f5 per cent of the total of 10 y. This statistical treatment of the data was carried through to show the inherent limitations of the technique. In our opinion, however, the method appears to be useful in clinical investigation.

Normally, red blood cell zinc constitutes 75 per cent, white blood cells 3 per cent, and plasma 22 per cent of total whole blood zinc. The greater portion of whole blood zinc is red cell zinc. However, the individual leucocyte contains about 25 times as much zinc as the individual erythro- cyte.

The data available at present do not disclose the possible differences in the zinc content of the various classes of leucocytes. Such studies must await the development of techniques for obtaining pure specimens of the lymphoid and myelogenous series of white cells.

The findings presented suggest that zinc is a physiological constituent of blood, and its individual variations in concentration follow the mathemati- cal pattern of commonly observed biological distribution phenomena.

The normal values here reported will serve as a base for comparison of the results of studies of blood zinc levels in the blood dyscrasias, now in progress.

Inasmuch as these studies employ the combined use of the chemical analyses and a radioactive tracer, Zn ‘j5, the data herein reported will serve as a basis for the study of the utilization of zinc in hematopoiesis, and the transport and distribution of zinc in the normal state and in disease.

SUMMARY

1. The zinc content of normal human whole blood, plasma, leucocytes, and erythrocytes in males and females has been determined.

2. Red blood cell zinc constitutes 75 per cent, plasma zinc 22 per cent, and white cell zinc 3 per cent of whole blood zinc in normal human blood. The individual leucocytes contain about 25 times the amount of zinc found in erythrocytes.

3. Zinc concentration in blood and its components follows the distri- bution pattern of physiological norms.

We wish to acknowledge the technical assistance of Miss Mary L. Roney, Miss M. Elizabeth Hickey, and Miss Barbara M. Clapp.

BIBLIOGRAPHY

1. Delezenne, C., Ann. Inst. Pasteur, 33,68 (1919). 2. Lutz, R. E., J.Ind. Hyg., 8,177 (1926).

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Bert L. Vallee and John G. Gibson, 2ndLEUCOCYTES, AND ERYTHROCYTES

HUMAN WHOLE BLOOD, PLASMA, THE ZINC CONTENT OF NORMAL

1948, 176:445-457.J. Biol. Chem. 

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