CALCULATION OF HEMOGLOBIN FROM BLOOD SPECIFIC GRAVITIES

BY DONALD D. VAN SLYKE,* ROBERT A. PHILLIPS,t VINCENT P. DOLE, PAUL B. HAMILTON,1 REGINALD M. ARCHIBALD, AND JOHN PLAZIN*

(From the Hospital of The Rockefeller Institute for Medical Research, New York)

(Received for publication, July 18, 1949)

The specific gravity of the red blood cells (normally about 1.10) so greatly exceeds that of t.he plasma that the difference between the gravities of whole blood and plasma can serve as a measure of the cell content of the blood. By assuming a constant hemoglobin concentration in the cells, one can estimate also the blood hemoglobin concentration from the gravity values.

The calculation is based on two assumed constants; viz., the specific gravity of the red cells and the hemoglobin concentration of the red cells. Neither of these values is in fact constant. In some abnormal types of blood cells, hemoglobin concentration may fall 20 per cent below normal, with cell specific gravity showing a parallel decrease. However, changes in cell hemoglobin concentration and cell specific gravity parallel each other in such a way that their variations almost cancel each other in their effect on the relation of blood hemoglobin concentration to the concentra- tion calculated from the gravities of whole blood and plasma. In conse- quence the error, even in grossly abnormal blood, is estimated to be only about 0.15 gm. of hemoglobin per 100 ml. of blood greater than the error attributable to the limit of accuracy of the gravity method used.

Ashworth and Tigertt (1) first formulated the equation for calculating blood hemoglobin concentration from the specific gravities of whole blood and plasma. The present paper reports tests of the accuracy of gravity- calculated hemoglobin concentrations by comparison with hemoglobin values determined by precise gasometric methods in normal and abnormal human blood. The gravity determinations have been made by the copper sulfate method (2). A nomogram (Fig. 1) is presented for rapid calcula- tion of hemoglobin from the whole blood and plasma gravities.

Formul.ation of Equations for Hemoglobin Calculation

Let Ds, DC, and Do be the densities (D:) of whole blood, plasma, and red cells, respectively. Let G,, Gp, and Gc be the corresponding specific

* Present address, Brookhaven National Laboratory, Upton, New York. t Present address, United States Naval Medical Research Unit No. 3, care of the

American Embassy, Cairo, Egypt. $ Present address, Alfred I. du Pout Institute, Wilmington, Delaware.

349

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350 HEMOGLOBIN FBOM BLOOD SPECIFIC GRAVITIES

Line chapt forwalculating plasma proteins, hemoglobin and hematocpit from gravities of plasma and blood

34 10.0 33 32

31

1.030 !

9.5 9.0

6.5

Hemoglobin HematocFit (gm./lOOcc.)

W) (Ht.1 5

5 15

6

\

20 1

0 25

L 36

- 37 - 38 - 39 -1.040

- 41

- 42

- 43

- 44

- 45

- 46

23 6.0

L; - 62 63

64 - 65

- 66 - 67

2&65

23 i

: ii -1.070

- 71

70 I 72

73

- 74

- 75

gravities in terms of 0:. Let Vc be the volume of red cells in 1 volume of blood. Let HbB be the gm. of hemoglobin in 100 ml. of blood, and Hbc the hemoglobin in 100 ml. of red cells. The weight in gm. of 1 ml. of

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blood, which is the density, is the weight of the plasma plus the weight of the cells in 1 ml. of blood.

(1) DB = DcVC+DPO - vc)

Since Ge, Gc, and Gp all are the corresponding D values multiplied by the same factor (1.003 when t = 25’) the gravity (DE) values may be sub- stituted for densities 0: in all terms of Equation 1.

(2) GB = GcVc + CPU - vc)

whence

(3) v = GB - GP

C- Gc - GP

Blood Hb concentration is cell volume X cell Hb concentration.

GB - GP HbB=HbcVc=HbcX -

Gc - GP

Constants must be substituted for Hbc and Gc. Hbc is calculated as Hba/Vs with Hb, determined by chemical analysis and Vc by hematocrit under standard conditions. Gc is calculated by rearrangement of Equa- tion 2.

(5) GB - GP

Gc = GP + _____ VC

The V, for Equation 5 is determined by hematocrit. From data given in the experimental part, Hbc for normal human blood

cells is taken as 33.9 gm. per 100 ml., and the corresponding Gc as 1.0964. We have taken these as the constants. Substituting them for Hbc and Gc in Equation 4 gives

03 GB - GP HbI, = 33.9 X 1.0g64 _ G

P

Cause of Relative Non-Ej’ect of Cell Abnormalities on Calculation of Hemoglobin by Equation 6

As mentioned above, in blood with cells of abnormally low density deviations of Hbc from 33.9 and of Gc from 1.0964 parallel each other in such a manner that the changes nearly cancel each other, numerator and denominator of the fraction (33.9(G, - GP))/(1.0964 - GP) being changed in nearly the same proportion by parallel deviations of the two constants.

In the hypochromic bloods of Table II, in fact, the correlation between abnormally low Gc and the error in gravity-calculated Hb, is so slight that

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352 HEbfOGLOBIN FROW BLOOD SPECIFIC GRAVITIES

it is not statistically definite, although in several bloods Gc is below 1.088 and Hbc below 29 gm. per 100 ml.

TABLE I

Data from Twenty Normal Men

Subject No. G

6 602 275 47.0 15.99 971 34.0 7 592 265 48.0 15.99 946 33.3 8 585 263 46.2 15.92 960 34.5 9 595 270 48.0 15.84 949 33.0

10 597 285 47.0 15.82 949 33.7

11 590, 265 46.0 15.78 12 5881 263 47.3 15.68 13 595! 277 48.0 15.63 14 582 267 46.0 15.47 15 588 265 46.4 15.46

16 587 277 45.0 15.12 966 33.6 17 582 273 44.0 15.12 976 34.3 18 568 250 44.3 15.11 968 34.1 19 570 257 45.0 15.03 952 33.4 20 577 267 45.0 15.01 956 33.3

Mean S.U. from mean

Maximum + devia- tion from HbOz

Maximum minus deviation from HbOz

Directly measured

Hemat-

:~*T&ole CP, writ, centn-

plasma fuged gravity gravity cf@Er

blood

Dz: D;; ml.

1.0635 1.0275 51.2 625 262 48.0 618 265 49.0 605 277 48.4 615 287 48.0

-____- 1.0595 1.0269 46.9

t0.OO21f0.OO09f1.8

C

-

1

-

Z

-

HbO, blood Hp

apacity

E 2:

17.56 17.46 17.10 16.46 16.35

15.90 t1 .oo

--

3

Calculated

1.0978 34.3 1018 36.4

985 35.6 955 34.0 970 34.0

971 34.3 9501 33.2 939’ 32.5

1.0964 33.9 zO.OO18i

i

E

/

_

Whole blood

[b from GB and GP

BY lquatior

6

% E 17.66 17.53 17.12 16.18 16.42

I p. gcr 100 ml.

+o. 10

f0.07 +0.02 -0.28 +0.07

16.09 +0.10 15.86 -0.13 15.58 -0.34 15.88 +0.04 15.58 -0.24

15.76 -0.02 15.72 +0.04 15.69 +0.06 15.32 -0.15 15.66 +0.20

15.30 15.16 15.10 15.01 15.08

+0.18 +0.04 -0.01 -0.02 +0.07

-0.01 f0.15 +0.20

-0.34

15.89

D 1

eviation from HbOe

Estimation of Hemoglobin from Spetifi Gravity of Whole Blood Alone

In blood with plasma gravities within the normal range 1.027 f 0.002 (Table I) one can, without much increase in error, estimate Hb, from Gs

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TABLE II

Data from Women Rejected As Blood Donors, Apparently Healthy But with Subnormab Hemoglobin

Directly measured

Subject No.

6 528 265 38.7 12.48 944 32.2 12.75 $0.27 7 528 275 40.0 12.50 908 31.2 12.45 -0.05 8 542 290 42.7 12.28 880 28.8 12.68 f0.40 9 480 260 36.1 10.24 869 28.4 10.59 +0.25

10 504 270 41.2 11.64 838 28.2 11.43 -0.21

11 12 13 14 15

16 17

Mean s.D. from mean Maximum + deviation

from HbCO Maximum minus devi-

ation from HbCO

T

Hemat. omit,

bzid p%L Yiz- pecific specific ravity gravity pegOC

blodd

I- Whole blood

-____

ID:: 0;: ml.

.0525 1.0255 39.9 510 265 37.6 506 275 37.5 503 255 37.0 508 275 37.0

E c:

12.96 11.65 11.50 11.79 11.76

-~

0:: Et?

.0932 32.5 917 31.0 891 30.7 925 31.9 895 31.8

BY DWir- “g- tion

from 6 HbCO

-- bn.w mm. P@ 100 ml. 100 ml.

12.91 -0.05 11.88 +0.23 11.37 -0.13 11.86 +0.07 11.46 -0.30

500 245 36.5 11.65 940 31.9 12.02 +0.37 545 270 41.5 12.96 933 31.2 13.43 +0.47 522 260 43.0 12.88 869 30.0 12.62 -0.26 520 255 38.5 12.63 944 32.8 12.67 $0.04 508 250 38.4 12.54 920 32.6 12.22 -0.32

524 275 39.7 500 265 37.7

---

.0515 1.0265 39.0

12.25 11.13

904 30.8 889 29.6

--

.0906 30.9

12.25 f0.00 11.40 +0.27

12.05 12.12 +o.os ltO.26 +0.47

-

T calculated

-0.32

alone, assuming that the plasma has the mean normal gravity, GP = 1.0269. Equation 6 then simplifies to

(7) Hbr, = 480(GB - 1.0269)

The maximal error in such bloods, introduced by a deviation of f0.002 from the assumed Gp of 1.0269, is about ~tO.5 gm. of Hb per 100 ml. of blood, or 3 per cent of normal Hbe. The data in Table IV indicate that

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354 HEMOGLOBIN FROM BLOOD SPECIFIC GRAVITIES

the observed error is of this order in normal bloods, and in simple anemia, due to iron deficit or blood loss, of the type of the blood donors in Table II. When varied pathological conditions ca.use wide fluctuations in plasma protein concentration and Gp, however (last column, Table IV), greater errors can arise in Hb, estimations made from Gs alone. Each gm. per 100 ml. of plasma by which the plasma proteins deviate above or below 7.4 gm. per 100 ml. causes a plus error in the same direction of about 0.7 gm. per 100 ml. of blood in the hemoglobin estimated by Equation 7. The latter can be used to follow hemoglobin changes in subjects that have been found to have plasma gravities within the range 1.025 to 1.029, and such use permits following Hb changes with finger blood. The extent of error that could arise, however, from undetected abnormality in plasma gravity is shown by a case with 12.7 gm. of plasma protein per 100 ml. reported by Atchley, Bacon, Curran, and David (3), in which the Hb, calculated by Equation 7 was 4.7 gm. per 100 ml. of blood too high. Forty-nine other cases reported by Atchley et al. (3) from a general medical ward showed a distribution of error (estimated as the deviation of Hbs by Equation 7 from Hbe by Equation 6) similar to that of the last column of Table IV, 94 per cent of the cases showing an error of less than 1.5 gm. of Hb per 100 ml.

Calculation of Hematocrit Values from G, and Gp

In Ashworth’s (1, 4) papers, and in preliminary reports by the authors (5), Equation 3 with 1.097 as constant value of G, was given as a means for approximate calculation of Vc from Gg and Gp, although it was recog- nized that the gravity-calculated Vc showed, from hematocrit-determined Ti,, percentage deviations that were much greater, especially in patho- logical blood, than the deviations between gravity-calculated and chemi- cally determined Hb, values. The reason for such Vc deviation is obvious from comparison of Equations 3 and 6. If the cell gravity deviates from the assumed 1.0964, its effect on gravity-calculated Vc is proportional to the change in the factor l/(Gc - GP), and is not canceled, as in Equation 6, by parallel change of a factor in the numerator. For example, in the blood of lowest Gc in Table II, Subject 10, the gravity-calculated Hb, agrees with that gasometrically determined within the limits of error, while the Vc value calculated by Equation 3 is 0.339, 18 per cent below 0.412 obtained by hematocrit. In normal blood Trc can be calculated by Equation 3 within ~0.02 (2 ml. of cells per 100 ml. of blood) of the hemat- ocrit-determined value, a =t5 per cent maximal error, but the error in- creases so greatly, even in the moderately abnormal blood of subject,s of the type in Table II, that calculation of Vc from gravities cannot be recom- mended, except in blood with normal cells.

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EXPERIMENTAL

Blood was drawn with syringes from the arm vein and transferred to tubes containing 0.2 mg. of heparin per ml. of blood.

The volume of packed cells per 100 ml. (190 V,) was determined by the conventional technique for clinical hematocrit values by use of Wintrobe (6) tubes. The blood was centrifuged at 3090 R.P.M. for 60 minutes, the center of each centrifuge tube being 18 cm. from the axis. (When a cen- trifuge was tried in which the centers of the tubes were only 9 cm. from the axis, 3900 R.P.M. for an hour gave with normal blood 51 ml. of cells per 100 ml. of blood, while the centrifuge with an 18 cm. axis gave 47.) The cell column obtained by the conventional technique contains, ac- cording to Gregersen and Schiro (7), about 7 per cent of its volume of in- separable plasma. The true cell volume would therefore be somewhat lower than the measured volume, and the true normal mean cell gravity somewhat higher than the 1.0964 obtained in Table I. The cell gravities, Gc, calculated in Tables I, II, and III, are the gravities of the packed red cells plus adherent plasma. If, applying Equation 5 to the mean normal values of Table I, one decreases Vc by 7 per cent, the value obtained for average normal G, becomes 1.1013 instead of 1.0964, and the Hb concen- tration of the cells would be 36.3 instead of 33.9 gm. per 100 ml.

Blood hemoglobin concentrations were determined gasometrically. The hemoglobin values of Table I were obtained by a modification of Van Slyke and Neill’s (8) oxygen capacity method, with details to obtain max- imal accuracy. Of each blood, soon after drawing, a portion of about 10 ml. was placed in a separatory funnel of 250 ml. capacity, and aerated by rotation as described by Stadie (9). After about 20 minutes 1 ml. was transferred to the Van Slyke-Neil1 manometric apparatus and the 02 + CO content was determined. Both gases, mixed with the CO2 and Nz, were set free from the blood by ferricyanide, the CO2 was absorbed, and the residual O2 + CO + Nz was measured. From the gas thus measured the sum of O2 + Nz physically dissolved in blood saturated with air at the observed temperature and barometric pressure was estimated from the Nz and O2 solubilities in blood ((10, 11) ; see also Fig. 48 (12)) in order to obtain the O2 + CO bound by the hemoglobin. This procedure was used instead of determination of the O2 alone by absorption from the ex- tracted gases with Naz’&04 solution, because the latter procedure fails to include the small amounts of CO, in the neighborhood of 0.3 volume per cent, bound as HbCO that appear to be present in normal blood regularly, at least in New York. This normal blood CO is increased by smoking, which may raise it to as high as 1 volume per cent. Only a small part of such CO is displaced by O2 in the process of aerating. The mean oxygen

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356 HEMOGLOBIN FROM BLOOD SPECIFIC GRAVITIES

capacity yielded by our results, 21.6 volumes per cent, is higher than the 20.7 previously estimated from oxygen capacity determinations ((12) p. 662) from normal men.

The determinations were done in duplicate or triplicate. The aeration in the Stadie rotator was continued during the entire period of the analy- ses, usually 40 to 60 minutes, so that any changes that might occur, such as formation of methemoglobin or transformation of methemoglobin pres- ent in vivo to active hemoglobin in vitro, might be detected. The accepted results were those in which consecutive analyses gave oxygen capacities within 0.05 volume per cent of each other. It has been shown that during the prolonged aeration any methemoglobin present is changed to active hemoglobin to such an extent that the oxygen capacity is 99 to 100 per cent of the total hemoglobin (13) in normal blood.

The hemoglobin values in Tables II and III were obtained by the “total hemoglobin” method of Van Slyke et al. (13) ; NazSz04 is added to reduce any methemoglobin present and the carbon monoxide-binding capacity of the Hb is determined as a measure of the hemoglobin. The total hemo- globin is designated as HbCO in Tables II and III.

Specijic gravities of whole blood and plasma were determined by the cop- per sulfate method previously described (2).

The values for cell gravities and cell hemoglobin concentration are cal- culated in Table I to obtain from normal blood the constants for Equation 6. In Tables II and III they are included to indicate the nature of cell abnormalities in the bloods examined. It will be noted that the group in Table II, although composed of women apparently healthy except for mild anemia, showed more marked tendency to low blood cell gravity and cell hemoglobin than did the patients of Table III. In Table IV are sum- marized the results which have already been discussed in connection with Equation 7.

Accuracy of Calculation of Blood Hemoglobin Concentration from Gravities of Whole Blood and Plasma

The results in Tables II and III indicate that, in patients with various types of anemia, the maximal deviation of Hb, estimated from gravities measured by the copper sulfate method, from Hb gasometrically deter- mined, is about f0.6 gm. per 100 ml. of blood; this figure is indicated both by the maximal observed deviations and by estimation as twice the stand- ard deviation. If 0.1 gm. per 100 ml. is allowed as the error of the gaso- metric determination, the maximal error of the gravity-determined Hb in pathological bloods of the types studied may be estimated as about ~~0.5 gm. per 100 ml., or 3 per cent of the normal hemoglobin concentration.

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TABLE III

Blood from &iiscellaneous Hospital Patients on Medical Service

Patient NO.

1* 1.0448 2 440 3* 468 4 495 5 448

%

1.0285 273 268 290 249

24.0 24.2 28.9 31.0 28.0

%i s:

8.25 8.43 9.50

10.32 9.59

1.0964 963 960 951 960

6t 289 225 9.2 3.28 920

7$ 589 313 42.8 14.17 958 8 389 243 20.2 6.93 965 9 485 249 33.2 11.03 960

10 465 250 30.4 9.86 957

11 12 13 14

1%

520 268 36.3 12.81 962 501 255 35.0 11.85 958 485 290 29.2 9.90 958 466 228 32.8 11.48 954 543 295 37.5 12.45 956

161 640 336 48.6 16.31 961 17 623 292 49.6 16.50 959 18 545 259 40.9 13.85 958 19 565 291 41.0 13.56 959 20 585 290 44.2 14.96 957

21 425 253 24.5 7.94 955 22 531 290 36.0 12.25 959 23 482 267 31.2 10.66 960 24 470 244 32.0 11.05 950 25 532 290 36.4 12.34 955

26 515 279

2711 555 270

WI 531 247

2911 511 260 ‘al 465 273 I -

34.9 11.70 41.6 13.75 40.0 13.69 36.1 12.13 28.4 9.66

955 955 957 955 949

GB

D::

-

-

Directly measured

GP

-

< Kemato- xit, cen- trifuged cells per

%z

-_

ml.

%z3 Hb by CO capacity

,

--

-

Calculated

CdlS

GC by.

EquSstron

Du” %i?%: 5% G E 2:

34.4 8.02 -0.23 34.8 8.19 -0.24 32.9 9.74 +0.24 34.6 10.31 -0.01 34.2 9.43 -0.16

35.7 2.94 33.1 14.37 34.3 6.86 33.2 11.18 32.4 10.21

-0.34 +0.20 -0.07 +0.15 +0.35

35.3 12.28 -0.53 33.8 11.76 -0.09 33.9 9.82 -0.08 35.0 10.96 -0.52 33.2 12.57 +0.12

33.6 16.41 33.3 16.70 33.9 13.76 33.1 13.81 33.8 14.82

+0.11 -to.20 -0.11 1-0.25 -0.14

32.4 8.20 +0.26 34.0 12.12 -0.13 34.2 10.52 -0.14 34.5 10.64 -0.41 33.9 12.17 -0.17

33.5 11.68 -0.02 33.1 13.92 -to.17 34.2 13.43 -0.26 33.6 12.09 -0.04 34.0 9.42 -0.24

-

-

Whole blood

Eb from GE and GP

BY Eq”riol

-

I 0

--

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358 HEMOGLOBIN FROM BLOOD SPECIFIC GRAVITIES

TABLE III-Concluded

r Directly measured Calculated -

t

--

_-

,

CdlS

Patient No. Hemato- wit, cen- trifuged cells per loo ml. blood

E3 Ib by CO :apacity

II Hb from GE and GP

318 570 3217 585

338 572 34ll 545 35ll 554

0%

262 268 260 266 260

44.2 45.9 44.9 40.1 42.1

E E

14.77 15.29 14.97 13.41 13.64

368 571 260 44.5 14.87 37 473 297 26.6 9.00 38 428 280 22.0. 7.42 39 421 242 25.1 7.93 40 470 271 29.0 9.97

41

428 435 44 45

526 290 35.2 11.64 485 274 31.1 10.41 389 273 17.0 6.41 562 269 42.2 13.96 569 266 43.7 14.79

46 509 252 36.3 47 473 258 30.9

12.37 9.81

GB GP

0:: Pd.

GC

by.

Y? kviation

from HbCO

952 959 955 962 959

F 2: E 2; %%

33.3 14.74 -0.03 33.3 15.45 +0.16 33.2 15.02 +0.05 33.4 13.55 +0.14 32.2 14.15 $0.51

958 33.4 14.95 +0.08 959 33.8 8.95 -0.05 952 33.7 7.34 -0.08 954 31.6 8.41 +0.48 957 34.4 9.73 -0.24

960 952 956 963 960

11.87 +0.23 10.36 -0.05

5.69 -0.72 14.29 +0.33 14.71 -0.08

960 953

33.1 33.3 37.7 33.1 33.9

33.9 31.7

12.24 -0.13 10.32 t-o.51

Whole blood

Mean deviation from HbCO. . S.D. from mean deviation. . . . . . . . Maximum +deviation from HbCO. . . . . . . .

“ minus deviation from HbCO. . . . . . . . . .

* Pernicious anemia. t Advanced nephritis; deficit of both hemoglobin and plasma protein. $ Myeloma; high plasma proteins. 0 Hodgkin’s disease. 11 Cirrhosis of the liver. 7 Hepatitis.

-

-0.02 f0.26 +0.51 -0.72

The error is about the same in blood with low as in blood with high hemoglobin concentration; it is an error constant in gm. per 100 ml. rather than in percentage of the hemoglobin present.

An important part of the error in hemoglobin calculated by Equation 6 from gravities in Tables I, II, and III is attributable to the limit of error of

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359

the gravity measurements by the copper sulfate method. The greatest error in Hb occurs when errors in GB and GP occur in opposite directions. If a plus-minus error in GE exceeding the standard error1 occurs once in 3 times, a plus error equal to the standard error occurs once in 6 times. Simi- larly a minus error in Gp equal to the standard error occurs once in 6 times. Simultaneous occurrence of the plus error in GB and the minus error in Gp would occur once in 36 times, and the reverse combination also occurs once in 36, so that errors in Hb from gravity errors exceeding their standard de- viations, and in opposite directions, would occur once in 18 times. If the “maximal” error is taken as one that would be exceeded once in 20 times, the maximal error of Hb calculated by Equation 6 would therefore approx-

TABLE IV Error of Hb Calculated from Gs Alone As Hb = .@O(GB - 1.0269)

Sourceofdata.. ........................................ Type of subjects .........................................

Total No. of subjects.. . . . . . _. . . .

Subjects with error under 0.5 gm. ‘I “ ‘I “ 1.0 “ “ “ “ ‘I 1 5 “ “ “ I< “ 2 0 “

Table I Normal men

20

Table II Rejected

wonvor; blood

17

90 (Is)* 94 (16) 100 (20) 100 (17)

m$lp” mlbpy

Mean error.............................. $0.03 -0.01 s.n. of error from mean. ~1~0.27 ztO.28

Table III Hospital

patients

per cecrt

62 (29) 81 (39) 92 (‘w

100 (47)

.m~p

+0.05 9~0.76

* The figures in parentheses denote the number of subjects.

imate (slightly exceed) the summated effect of opposite errors in Gp and GLB equal to their standard deviations, which have been found to be f0.0003 and f0.0004 gravity unit respectively (2). The error in Hbe caused by errors of JrO.0004 in G, and -0.0003 in GP is approximately 0.3 gm. of Hb per 100 ml. of blood, which is about the maximal error found in gravity analyses of normal blood (Table I). It appears that nearly all of the error of HbB calculated from GB and GP in normal subjects (Table I) is attribut- able to the error of the copper sulfate gravity method, and about half the error in the abnormal bloods (Tables II and III). A gravity method free

r Standard error = standard deviation of gravity by CuSO4 method from gravity by pycnometer. By the probability equation, 0.317, or l/3.15, of the gravities by CuSOd may be expected to deviate from pycnometric gravities by more than the standard error.

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360 HEMOGLOBIN FROM BLOOD SPECIFIC GRAVITIES

of error could be expected to reduce the “maximal” Hb error in normal blood to nearly zero, and in pathological blood to 0.2 or 0.3 gm. per 100 ml.

In Fig. 1, for convenience in rapid calculation, is a nomogram prepared by standard methods (14) for calculation of plasma protein and blood hemoglobin concentrations from gravities. The plasma values are calcu- lated by Equation 3 of the preceding paper (15), the hematocrit and hemo- globin values by Equations 3 and 6 respectively of this paper.

SUMMARY

In twenty normal and 64 abnormal, mostly anemic, heparinized human bloods, specific gravities were determined by the copper sulfate method. From specific gravities of the whole blood and plasma, blood hemoglobin concentrations were calculated by the formula of Ashworth and Tigertt, and were compared with hemoglobin concentrations determined by pre- cise gasometric methods.

The standard deviation of the gravity-determined from gasometric hemoglobin was f0.15 gm. per 100 ml. of blood in the normal bloods, and 10.26 gm. in the abnormal.

A portion of the deviation equal to nearly all that observed in the normal bloods is estimated to be attributable to the limit of accuracy of the copper sulfate gravity method.

A nomogram is presented for rapid calculation of plasma protein and blood hemoglobin concentrations from specific gravities.

BIBLIOGRAPHY

1. Ashworth, C. T., and Tigertt, W. D., J. Lab. and Clin. Med., 26, 1545 (194041). 2. Phillips, It. A., Van Slyke, D. D., Hamilton, P. B., Dole, V. P., Emerson, K., Jr.,

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M. Archibald and John PlazinVincent P. Dole, Paul B. Hamilton, Reginald

Donald D. Van Slyke, Robert A. Phillips,FROM BLOOD SPECIFIC GRAVITIES

CALCULATION OF HEMOGLOBIN

1950, 183:349-360.J. Biol. Chem. 

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