THE EFFECT OF PHOSPHATE ON THE SOLUBILITY OF

CALCIUM CARBONATE AND OF BICARBONATE ON

THE SOLUBILITY OF CALCIUM AND MAGNESIUM
PHOSPHATES
BY ISIDOR GREENWALD
(From the Department of Chemistry, New York University College of Medicine
New York)
(Received for publication, August 30, 1945)

In 1925, Holt, La Mer, and Chown (1) reported that the apparent solu-
bility product [Ca++13[P04=J2was greater in a solution of inorganic salts
approximating the composition of serum than it was in solutions of sodium

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chloride of the same ionic strength. Very shortly thereafter, Sendroy and
Hastings (2) recalculated values for pK’, Ca,(PO& from the data of Holt,
La Mer, and Chown and also reported extensive data of their own. Ac-
cording to Sendroy and Hastings, the solubility of calcium phosphate was
the same in “serum salt” solutions as in sodium chloride solutians and was
between those values calculated from the data of Holt, La Mer, and Chown
for the solubility in sodium chloride and in “serum salt” solutions. Sen-
droy and Hastings suggested (p. 821) that the latter difference might be
due to the “use of what may be uncertain values for the first or the third
stoichiometric dissociation constant of phosphoric acid at such low pcllH+,”
but they seem to have made no attempt to calculate whether or not any
conceivable error in these dissociation constants could have accounted for
the observed lOO-fold change in the apparent solubility product. We have
made such calculations and do not find that any conceivable error in any,
or all, of the values for the dissociation constants of phosphoric acid could
produce any significantly different effect upon the ratio of POh’ to total
phosphorus at pH 5.2 or 7.7, which represent the extreme values in the two
sets of experiments by Holt, La Mer, and Chown.
The “serum salt” solutions employed by Holt, La Mer, and Chown con-
tained, besidescalcium hydroxide and phosphoric acid, sodium, potassium,
and magnesium chlorides and sodium bicarbonate. We have since
reported evidence for the existence, in solution, of undissociated calcium
and magnesium carbonate, bicarbonate, and phosphate (3, 4). However,
the amounts of such complexes that could have been present could account
for only a small part of the observed greater solubility in “serum salt”
compared with that in sodium chloride solutions.
The composition of the “serum salt” solutions used by Sendroy and Hast-
ings is not given. They do not seem to have contained magnesium.
697
698 SOLUBILITY OF CA AND MG PHOSPHATES

Moreover, it is stated ((2) p. 799) that, in the “sodium chloride” series, the
ionic strength up to 0.03 M was furnished by sodium bicarbonate and only
above that by sodium chloride. This suggested that the greater solubility
in “serum salt” solutions observed by Holt, La Mer, and Chown might
have been due to the bicarbonate. If this were the case, no difference in
solubility was to be expected in the two series of experiments by Sendroy
and Hastings, since both contained bicarbonate.
It was obviously desirable to repeat the work, with sodium chloride-
and bicarbonate-containing solutions at approximately the same pH.

TABLE I
Solubility of Calcium Phosphate in Sodium Chloride Solutions (p * 0.155)

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-
Exper;mt -Log [Ca++13[HPOhlZ
PH Ca P CaHPO,
[aH+]z

moles x 106 moles x 106 nzoles x 106

a34* 6.80 710 780 8 2.67

836* 7.05 477 622 6 2.68
837* 7.18 423 597 6 2.53
838* 7.33 292 523 4 2.75
839* 8.02 106 432 1 2.68
931t 6.95 730 345 5 2.90
932t 7.02 770 330 5 2.68
9331 6.80 784 430 5 3.05
934t 6.70 1135 525 10 2.80
935t 6.68 1300 670 11 2.38

Average, calculated from values for [Ca++]3[HP04=]2/[aH+]2. . . . . 2.678 f 0.067

*Equilibrated for 29 days.
t Equilibrated for 60 days.

EXPERIMENTAL

The general conduct of the experiments and the methods of analysis

mere those reported in previous work (3-5). The bicarbonate content was
determined by adding an excess of standard acid, boiling to expel COZ,
titrating the excess acid, and correcting for the phosphate present and, in
the more alkaline solutions, for the amount of carbonate. Total carbon
dioxide was then calculated.
The mixtures were kept at room temperature, about 22”, for the periods
indicated in Tables I to IV, and were shaken several times almost every
day.
The following constants were used: for H&OS, pKi = 6.34 and pKi =
10.10; for HsP04, pKz’ = 6.78; for -log[Ca++][HCO3]/[CaHC03+] =
0.81; for -log[Ca++][CO,=]/[CaC03] = 3.00; for -log [Ca++][HPO*=]/
[CaHPOd] = 1.50; and 0.77, 2.31, and 1.50 for the negative logarithms of
 I. GREENWALD 699

the instability constants of the corresponding magnesium complexes.

For the calculations from the data of Sendroy and Hastings and for those
calculated by them from the data of Halt, La Mer, and Chown, we em-
ployed the constants used by Sendroy and Hastings for H.jZOs and H3P04,
the instability constants given above, and -log[Ca++]3[HPO~=]2/[aH+]2
= 4.11 (5).

TABLE II
Solubility of Calcium Phosphate and Calcium Carbonate in Solutions Containing Both
HPO,= and HCO,- (M = 0.155); Equilibrated 20 to 30 days

Total Calcium as
- - - -

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j

PH
CC+ P co2

8
“,
em
_ u” 6
moles moles moles moles moles moles moles moles
x 106 x 100 x 104 x 10” x 106 x 100 x 106 x lob
6.90 1485 1790 258 51 4 9 20 490 1.43
7.00 1250 1390 260 51 5 7 19 384 1.31
7.15 1180 1360 255 40 5 6 28 292 1.37
7.32 848 1190 251 31 7 5 28 267 1.25
7.50 676 1090 251 24 7 4 33 207 1.20
7.85 504 1010 248 11 9 2 54 150 1.33
8.30 274 857 245 4 10 1 58 58 1.37
8.63 166 824 256 2 10 0 48 23 1.31
8.90 126 800 263 1 10 0 44 11 1.27
9.01 120 826 273 1 10 0 44 9 1.27
2.- -
-Log of average of K........................................... 1.33

Initial [Cal = 0.001 M; [P] = 0.00125 RI; [CO,] = 0.025 M. Solid phase as in experi-
ments of Table I. [Ca++] in Series A calculated from -log[Ca++]3[HP04=]2/[Ca-
(CaP04)2][aH+]2 = 2.678 (Table I). [Ca ++] in Series B calculated from pK,,CaCO,
= 8.007 (3). -Log [aH+] [Ca2~PO~~C03]/[Ca2~H.PO~~COz] = 8.3.

The initial concentrations of Ca and P were 1.00 mM in Experiments

834 to 839, 0 in Experiments 931 to 935 (Table I), and 1.25 rniM in all the
experiments of Table II. The solid phase, in the experiments of Tables I
and II, was provided by the addition of 250 mg. per liter of Preparation 147.
This was obtained by the slow addition (several days) of a liter of 0.02 M
Na2HP04 to about 3 liters of a solution containing 0.18 mole of NaHC03
700 SOLUBILITY OF CA AND MG PHOSPHATES

and 0.033 mole of CaCL. This material was one of several prepared by
various methods in an attempt to obtain a solid phase which might repre-
sent the actual solid phase in equilibrium with bicarbonate-phosphate solu-
tions. The attempt was unsuccessful, and no further report will be made.
The various preparations were crystalline, resembling CaHP04 in form,
but had varying compositions. The particular material used in all the
experiments reported in this paper contained, per 0.250 gm., 2.06 mM of
Ca, 1.19 mM of P, and 0.32 mM of COz. It did not remain crystalline, but
became amorphous and, as may be calculated from the data in Tables I
and II, the final composition of the solid phase varied, the ratio of calcium
to phosphorus increasing with the pH of the mixture.
It will be noted that, in Experiments Aq, Aa, and AZ, the final concen-

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trations of calcium and phosphorus were greater than the initial concen-
trations. The apparent greater solubility cannot, therefore, be due to
delayed precipitation.
DISCUSSION

It was quite apparent that the solubility of calcium phosphate was in-
creased by the presence of bicarbonate and that the solubility of calcium
carbonate was increased by the presence of phosphate.
An attempt was made to interpret the results in terms of complex forma-
tion. Inasmuch as it has already been shown (3, 4) that such solutions
seem to contain CaHC03+ and CaHPOd, it seemed possible that the latter
should dissociate H+ and that the remaining Cap04 might combine with
CaHCOs+ to form a complex, Caz.H.POd.COa. It was postulated that
the mixtures were actually saturated with either Ca(CaP04)2 (or some sub-
stance sufficiently like this to permit the use of the product [Ca++]3[HP0,=]2
/[aH+]” as the limiting factor) or with CaC03. Accordingly, the [Ca++]
was calculated from the previously determined values of [Ca++]z[HP04=]2/
[aH+12 (Table I), or of [Ca++][CO,=] (3) in sodium chloride solutions. The
lower value for [Ca++] was adopted as a first approximation. With the
previously established values for the instability constants of CaHP04,
CaHCOs+, and CaC03, first approximations to the concentrations of each
of these were calculated. The difference between the concentration of
total calcium and that of the sum of the ionic calcium and the three pre-
viously postulated complexes was regarded as being due to the “new com-
plex.” After the concentration of bound phosphorus and bound carbon
dioxide had been subtracted from the respective totals, the calculation
of the concentration of the different forms of calcium was repeated until
there was no appreciable change. The values for -log[Ca*]2[HP04=]
[HC03-]/[Caz.H.P04.COB] [aH+] thus obtained were quite constant over
a considerable range of pH, but showed a tendency to rise above pH 7.85.
 I. GREENWALD 701

This change seemed to be greater than could be accounted for by any

error in analytical technique or in the choice of constants. It was recalled
that CaHCOs+ dissociates H+ and that -log[CaCOd[H+]/[CaHCOz-] =
7.9 (3). It was supposed that Caz.H.P04.COs might similarly dissociate
Hf. Calculations, the results of which are summarized in the last three
columns of Table II, indicate that this hypothesis may be correct and that,
if so, the mid-point of the dissociation is at pH 8.3. If this a.ssumption is
made, the values for -log[Ca++]2[HP04=][HC0~]/[Ca~~H~P0~~CO~][aH+]
are reasonably constant over the whole range of pH, 6.90 to 9.01.
This method of calculation was applied to the data of Sendroy and Hast-
ings obtained at 38“. For - log[Ca]3[HPOh]2/[aH+]2, the value 4.11 pre-
viously (5) calculated from the data of Holt, La Mer, and Chown, as re-

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calculated by Sendroy and Hastings, was employed. The values for in-
stability constants of CaHP04, CaHC03+, and CaC03 were those obtained
at room temperatures. This may have introduced a slight error. Be-
cause the values for the instability constant showed no tendency to change
as the pH varied between 7.18 and 7.64 and because, even at the latter pH,
the concentration of (Ca2.POJ*COa)- would still be expected to be small,
the formation of the latter was neglected. Although the values for the
concentration of the “new complex” are obtained by difference and reflect
all the errors involved in the determination of the concentrations of three
different constituents and the calculation of the concentrations of three
others, the values for the instability constants of this hypothetical new com-
plex show, for ten of the eleven experiments accepted by Sendroy and
Hastings,’ rather less variability than do those for the product [Ca++13
[Poe’].” The values for the means are 5.75 k 0.165 X lo-? for the former,
and 3.69 f 0.153 X 1O-27 for the latter.
In the foregoing calculations, we have assumed that the “serum salt”
solutions employed by Sendroy and Hastings did not contain magnesium.
It is obvious from a comparison of their Table VIII with their Table I
that this was certainly true of Experiments 7’ and 7A’, which appear in
both the sodium chloride and “serum salt” series. Even if magnesium
had originally been added, it is possible, even probable, that a considerable
portion would have been precipitated, or adsorbed, onto the large amounts
of solid calcium phosphate and carbonate t.hat were added (1 gm. of each
per 100 ml.). The mixtures employed by Holt, La Mer, and Chown did
contain magnesium and, since, at the most, only 8.2 mM of phosphate were
precipitated per liter, and no solid phase was added as such, probably very
little magnesium was precipitated with the calcium phosphate. Therefore,
1 Experiment 15 was omitted because an accurate value for the instability con-
stant cannot be calculated because the concentration of phosphorus is so low that
five-sixths of it is bound as complex, leaving only 1OF M .present as free phosphate.
702 SOLUBILITY OF CA AND MG PHOSPHATES

there must have been present, in solution, MgHCOs+, MgC03, and Mg-
HPOI. There was also the possibility of the formation of a complex similar
to that postulated for calcium and of mixed complexes as well.
The former of these possibilities was investigated by the determination
of the solubility of magnesium phosphate in the presence and absence of
bicarbonate. The apparent solubility product of MgHP04 had previously
been determined by Tabor and Hastings (6) and the negative logarithm
found to be 4.46 f 0.03,2 at CL * 0.16 and at 38”, over a rather narrow
range of pH and of phosphate concentration. As may be seen from Tables
III and IV, our values over a far greater range of pH, phosphate, and mag-
nesium concentrations are similar to that of Tabor and Hastings, and are
almost identical in the presence and in the absence of bicarbonate, 4.225 GZ

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0.067 and 4.12 f 0.03, respectively. The failure to find any such differ-
ence as was observed with calcium phosphate may appear rather surpris-
ing, but it must be remembered that magnesium carbonate and phosphate
differ in other respects from the corresponding calcium compounds. Mag-
nesium carbonate dissociates in water to form basic salts and even the hy-
droxide, while calcium carbonate does not. On the other hand, the ap-
parent solubility product for MgHP04 is constant even up to pH 7.8
(Table III) or pH 8.0 (Table IV), whereas with calcium the corresponding
product fails at least 2 pH units below this and the precipitate obtained
contains calcium and phosphorus in the atomic ratio of 1.5, or more, to 1.
In calculations from the data of Holt, La Mer, and Chown, allowance
was made for the formation of MgHCOs+, MgC03, and MgHPO+ but
not for the existence of mixed complexes containing both magnesium and
calcium. The apparent instability constants thus found varied between
2.1 X lo+ and 4.7 X 10-2, with an average of 2.9 X 10-2. These are
somewhat smaller than in the case of the experiments of Sendroy and
Hastings. This difference may be the result of a systematic error due to
differences in technique in the two laboratories or may, indeed, indicate
the real existence of mixed complexes.
It may occur to the reader that the hypothesis developed in this paper
requires the assumption of t.he existence of five new substances and of five
instability constants. In a sense that is true. However, the existence of
three of these complexes and the validity of the corresponding constants
have been established in previous publications (3, 4), and the existence of
MgHP04 and the value of its instability constant have been verified by
Tabor and Hastings (6). Therefore, it is now necessary to assume only
the existence of a new pair of complexes, related to each other’as acid and
anion.
2 The calculation of the standard deviation is ours.
 I. GREENWALD 703

TABLE III
Xolubility of Magnesium Phosphate in Absence of Bicarbonate
T

Equilibrated PH Mg P pK’,MgHPOh

day.? moles x 103 moles x 103

1 6 7.77 9.88 8.42 4.26 151
2 6 7.81 10.65 9.10 4.20 148
3 6 7.40 20.7 9.24 3.97 163
4 10 7.43 13.1 11.1 4.08 128
5 11 7.11 14.5 12.8 4.04 124
6 11 6.90 14.8 12.7 4.09 129
7 11 6.91 18.5 12.25 4.01 136
8 11 6.82 19.4 11.55 4.05 138

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41 6 7.10 4.64 35.85 4.19 149
42 6 7.10 4.26 35.45 4.23 149
43 6 7.03 3.84 36.5 4.28 155
44 6 6.90 4.26 33.8 4.30 149
109B 15 6.33 51.0 12.25 3.90 149
109c 15 6.33 56.3 6.13 4.14 165

Average, calculated from average for ion product.. 4.12 f 0.03

TABLE IV
Solubility of Magnesium Phosphate in Presence of Bicarbonate
T
Experm Equili- P cot
brated PH Mg pK’,MgHPOa B x 103
.- I-
days de.7 x 10, :oles x 10: Jffl 01‘3s x 10:
1 28 6.78 57.0 2.43 23.2 4.41 157
2 28 6.30 52.9 3.60 33.2 4.42 156
4 32 6.40 40.8 2.85 T.31 4.54 121
5 32 6.88 42.0 2.00 17.25 4.47 125
6 35 6.80 41.3 2.18 14.0 4.47 122
11 11 s.00 9.38 7.94 51.6 4.39 146
12 11 7.80 14.0 7.90 38.8 4.21 147
13 20 7.69 10.1 9.10 30.3 4.28 135
14 20 7.40 12.3 10.8 10.4 4.16 125
21 19 7.30 8.86 8.34 52.0 4.44 141
22 14 7.22 10.1 9.17 41.3 4.45 136
23 14 7.12 10.75 9.86 31.7 4.41 130
24 14 6.80 14.6 11.9 14.4 4.18 125
51 14 7.32 10.1 40.0 50.7 3.77 166
52 14 7.20 7.93 38.5 51.5 3.99 157
53 14 7.22 6.13 35.2 50.5 4.11 158
54 14 7.20 5.92 33.7 63.5 4.05 157
-
Average, calculated from average for ion product. 4.225 f 0.067
704 SOLUBILITY OF CA AND MG PHOSPHATES

SUMMARY

The solubility of calcium carbonate at p * 0.155 is increased by the

presence of phosphate, and that of calcium phosphate is increased by the
presence of bicarbonate. Data are presented that indicate the existence
of the complexes (Caz.H.POa.COJ and (Caz.PO1.COs)-, with -log
[aH+][(Caz~PO~~COJ)-]/[Ca~~H~PO~~COa] = 8.3.
Over a considerable range of calcium, phosphorus, and hydrogen ion
concentration, the value for -log[Ca++]2[HP04=][HC0S-]/[Ca2.H.P04.
CO&aH+] = 1.33. The hypothesis is shown to be consistent with data
previously reported by others.
The apparent solubility product for MgHP04 has been determined over
a wide range of concentrations of magnesium, phosphate, and hydrogen

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ions and found to be the same in the presence, as in the absence, of bicar-
bonate.

BIBLIOGRAPHY

1. Holt, L. E., Jr., La Mer, V. K., and Chown, H. B., J. Biol. Chem., 64, 509 (1923).
2. Sendroy, J., Jr., and Hastings, A. B., J. Biol. Chem., 71, 797 (1926-27).
3. Greenwald, I., J. Biol. Chem., 141, 789 (1941).
4. Greenwald, I., Redish, J., and Kibrick, A. C., J. Biot. Chem., 136, 65 (1940).
5. Greenwald, I., J. Biol. Chem., 143, 703 (1942).
6. Tabor, H., and Hastings, A. B., J. BioZ. Chem., 148, 627 (1943).