Sie sind auf Seite 1von 11

THE EFFECT OF HEAT ON THE SOLUBILITY OF THE

CALCIUM AND PHOSPHORUS COMPOUNDS


IN MILK.
BY RAYMOND W. BELL.
(From the Department of Dairy Industry, Cornell University, Ithaca, N. Y.)

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


(Received for publication, April 6, 1925.)

INTRODUCTION.

In reviewing the literature on .the effect of heat on milk, it


appears that considerable difference of opinion still exists relative
to the probable effect of heat on t,he chemical and physical proper-
ties of the mineral salts of milk.
Some observers claim that the heating of milk causes part of the
soluble compounds containing calcium and phosphorus to change
to an insoluble condition and be precipitated, while others claim
that there is no noticeable change in the solubility of the com-
pounds. Not only is this found in the strictly chemical investiga-
tions of milk, but in the nutrition experiments conducted by Lane-
Claypon (1916), I>aniels and Loughlin (1920), and others on the
nutritive value of raw and boiled milks.
If a soluble calcium or phosphorus deficiency exists in heated
milk, the logical way to prove it is by chemical analysis, but on turn-
ing to the investigations along chemical lines, we are confronted
with marked differences in data and opinions with regard to the
influence of heat on the composition of the milk. The probable
reason for this undoubtedly lies in a lack of uniformity of methods
and milks used. Some used whole milk, others skim milk. The
age of the milk is of especial importance, the older milk having the
higher acid content, due to bacterial action. In filtration experi-
ments carried on in connection with this investigation, it wasfound
that as the acidity of the milk in the filtering apparatus increased,
due to natural souring, the amount of CaO and P,Ob in the filtrate
gradually approached a value close to that of the total CaO and
P,Os content of the fresh skim milk before starting to filter.
391
392 Cs and P Compounds in Milk

Methods.
Treatment of Milk.
Standardization of methods and elimination, as far as possible,
of all factors which might tend t,o produce a change in the milk,
ot,her than heat, arc of prime import.ance, so as to have the samples
on a uniform basis for comparison. With t,his in mind and to
overcome the objections previously discussed, t(he following general
methods were carried out,.

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


In all of the experiments performed, only fresh, raw, skim milk
was used. As soon as obtained, it was thoroughly mixed and
divided into two portions, one part going to the cooler to prevent
chemical change and the other taken to the laboratory for the
beginning of the experiment. Skim milk was used because the
fat of whole milk would clog the pores of the Pasteur-Chamber-
land filt,ers used and also, in the second part of the work, because
of the construction of the supercentrifuge.
In the work with the Pasteur-Chamberland filters, the milk was
heat,ed in a water bath in an open container. No account was
taken of the loss in volume due to evaporation. In the work with
the supercentrifuge, the milk was heated under a reflux condenser,
thus preventing a loss due to evaporation. In all cases, as soon
as t,he desired temperature for holding the milk had been reached,
t,he temperature of the water in the water bath was brought down
t.o that of the milk by the addition of cold water. All samples
were mainbained at the holding temperat.ure for 30 minutes. At
the end of that time, t,he samples heated in an open container
were cooled as soon as possible under the cold water tap. Those
samples in the second part of the work were cooled in the same
way except that t,he container was closed. In all cases, the milk
container in the wat,er bath rested on a wire support, thereby pro-
tecting the milk at the bottom of the container from the more
direct, heat near the source of application. Up to the time of
cooling, the milk was agitated only enough to insure an even dis-
tribution of heat. On cooling, however, the milk container was
continuously agitated while under the tap to hasten the cooling
process. This continued until t,he milk in t,he cont,ainer had
attained a temperat,ure of 20°C.
Raymond W. Bell 393
It can readily be seen that the time of heating to bring the milk
to the holding temperature increased as the t,emperature of the
holding became higher. The intensity of the heat applied was
regulated as much as possible so that t>he increased 1engt.h of time
t,o attain the higher temperature would have a definite ratio. For
instance, it would take 20 rninut’es to heat t,he milk to 180”F.,
25 minutes to at,tain 19O”F., 30 minutes to reach 200”F., etc.
It also holds that the higher t.he holding t,emperature, the longer
the t)ime necessary for cooling.

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


Filtration Methods.
Many of our ideas concerning the state in which the various
constituents of milk occur, are the result of filtration studies in
which the milk was filtered through earthenware filters. The
variety of individual filters used, influenced the results. Among
the more import,ant’ studies of this t,ype, that. of Rupp (1913) is
of special interest. He obtained a serum by filtering milk through
“a porous clay cell of a galvanic element,” using suction. He con-
cluded that pasteurizatjion for 30 minutes at 68°C. did not affect
the content of soluble calcium, magnesium, and phosphorus com-
pounds in the serum.
Probably the most extensive studies of the condition of the
various compounds in milk are those of Van Slykc and Rosworth
(1914). Others who have worked on the effect of heat on milk
are Palmer (1921-22), Palmer and Scott (1919), Jackson and
Rothera (1914), and Flohil (1911). Grosser (1919) hasshown that
raw milk gave an ultrafiltratc much richer in lime than did boiled
milk. In boiling, the Ca is bound to the milk colloid and remains
with the latt,cr on the ultrafilter.
The methods of study most, frequent,ly resorted to are dialysis
and. filtration. Dialysis studies at best are unsatisfactory when
working wit,h such a complex material as milk because of t,hc
danger of shifting the original state of equilibrium. On t,his
account, filtration met,hods seem more desirable. Much of the
work already report,ed is difficult to interpret because of the
apparent unknown variations in the porosity of the filt,ers used.
It therefore seemed desirable to attempt filt,rat.ion studies, using
filters the relative porosity of which was known.
394 Ca and P Compounds in Milk

Briefly stated, t,he process consisted in putting the milk into a


t,ubular chamber surrounding a Past.eur-Chamberland filtering
tube.. Pressure amount.ing to 50 t’o 60 pounds per square inch
was then applied by means of a pump, which forced air into the
chamber containing the milk and caused the soluble portion of the
milk to pass through the walls of the filter from the outside to the
inside of the filter t,ube. Any pressure greater than that mcn-
tioned did not seem to increase the speed of filtrat,ion.
The insoluble residue accumulated on the outside surface of the

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


filter tube, from which it was removed by light scraping with a
stiff brush and the use of running water forced from the inside to
t,he outside of t,he tube. Since the filter has the power of adsorb-
ing some of the constituent’s of the serum until a volume of 50
to 75 cc. has passed through, this amount should be discarded.
After this, the filtered serum was constant in composition. This
was the opinion of Rupp (1913), and it was verified in this work by
t,rials, the results of which are not given in the data here reported.
Before being placed in t,he apparatus for filtration, the milk
was cooled to 4OV. to prevent souring during the process of filtra-
Gon. No antiseptic for preserving was ever used. The filtering
apparatus was then placed in the cooler where a constant tempera-
ture of approximately 40”E’. was maintained. IXew candles were
used t)wo or three times before using them for experimental data.
In cleaning, the direct,ions as given by the company manufac-
turing tire candles were followed carefully.

Centrifugal Methods.

Milk is an aqueous solution of crystalloids (salts and milk sugar)


which contains the colloids, casein and albumin, and also an
,emulsion of fat,. If, according to Grosser (1919), the lime is
attached more firmly to the milk colloids in boiled milk than in
unboiled milk, then, provided a suitable means of separating the
milk colloids from the aqueous solution of crystalloids is found,
the lime and phosphorus cont,ent of the aqueous solution of crys-
talloids should be less in boiled milk than in raw milk from t,he
same source.
Therefore, in addition t,o the experimental met,hods already
described, it was decided to make this separation by means of a
high speed cent’rifuge. The supercent,rifuge was so constructed
Raymond W. Bell 395
that the milk could be passed cont,inuously through the rotating
member, where it was subjected to a force approximately 40,000’
times that of the force of gravity and was then continuously
discharged.
The samples of milk were obtained and prepared as described
previously. Blthough 120°F. is the temperature of minimum
viscosity for milk, it was t,hought, best to make the centrifuging
temperature for the raw and heated skim milk samples 70’F. which
is normal room temperature. 500 cc. of each kind of milk were

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


centrifuged each time. After about 300 cc. of the liquid had
entered the bowl, the bowl was full enough to overflow from the
discharge holes in the top into the cover. The liquid then flowed
from the cover into a suitable dry container.
At the end of the operation, the machine was allowed to stop,
the bowl was removed, and the deposited solid matter obtained by
scraping the contents into a t’ared porcelain dish. The solid
matter was dried and incinerated and the weight of the ash
obtained in each case. The ash was in turn analyzed for the
weight of lime (CaO) and phosphorus (P205) which it contained.
The liquid discharged from the cover (which will hereafter be
called the filtrate) was measured for the volume and 100 gm. of
it were ashed and analyzed for the weight of CaO and PzOa which
it contained. The volume of the filt.rate was in every case approxi-
mately 190 cc.
Analytical Methods.
The phosphorus analyses were made as recommended by the
Association of Official Agricultural Chemists.
The calcium analyses were made according to McCrudden
(1911-12).

RESULTS.

Filtration Experiments.
In order to test the variability of individual candles, a number of
tests were made in which duplicate samples of raw skim milk
were passed through different filters. From the results, as shown
in Table I, it was computed that there was a possible variation
between the candles of 0.0042 per cent in the determination of Ca
396 Ca and P Compounds in Milk
in the filtrate, and of 0.0038 per cent in the determination of
Pz05 from the same source.
Experiments were then carried on to see if the difference in
the percentage of Ca and PZO, in t,he filtrate from the raw and
heated samples from the same source would exceed this value.
The candles were used interchangeably, no means being taken to
distinguish one from the other in succeeding experiments. The

TABLE I.
Comparison Candles on Duplicute

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


of Runs of Ruw Skiln Milk.
- -
Run No. Chdle. Ca PP,OG Dii%er; in

per cent per cent per cent per cent


1 0.0265 0 0627
2 0.0264 -0.0001 0.0606 -0.0021

1 0.0253 0.0674
2 0.0249 -0.0004 0.0656 -0.0018

3 0.0280 0.0660
4 0.0318 JrO.0038 0.0686 +0.0017

5 0.0215 0.0646
6 0.0215 0.0000 0.0633 -0.0013

5 0.0254 0.0612
6 0.0254 0.0000 0.0609 -0.0003
- - -
Avera,qe......................... $0.0007 -0.0007
- - I

average difference between the percentage of Ca and P205 in the


filtrate from the raw and heated milk was obt,ained by determin-
ing the sum of the positive and the negative differences and
dividing t,hat difference by the total number of determinations
made at that particular temperature. For example, at 140°F.
the loss in the Ca content, of the filtrate from the skim milk, as a
result, of heating, exceeded the gain by an average of 0.0032 per
cent for six determinations. The loss in the Pn05 content of t,he
filtrates from t,he skim milk, due to heat,ing, exceeded the gain
by an average of 0.0018 per cent for six det.erminations.
While the results obtained with the samplesheated to tempera-
Raymond W. Bell 397
t,ures of 140-160°F. yielded results which perhaps fall within the
experimental error, the experiment,s with the higher t.emperatures
gave more marked differences. The great,est. variation between
any two of the candles used, when duplicates of raw skim milk
were filt,ered, was 0.0041 per cent. in the case of Ca and 0.0038
per cent in the case of P,Os. It appears t,hat the difference in the
percentage composition of Ca in the filt.rate from raw and heated
skim milk, and of P,O, from the same source and under the same
experimental conditions, does not, exceed the possible experimen-

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


tal error in the candles unless the milk has been heated for 30
minutes to 170°F. or above. Even at t,hese temperatures, the
difference is small.

TABLE II.

SummaryojLosses of Soluble Ca and PnOs Dueto Heating (Filtration Method).

Temperature to which Average loss in PzOa i o


heated for 30 min. Average loss in Ca in filtrate. filtrate.

“F. per cent per cent


140 0.0032 0.0018
150 0.0011 0.0024
160 0.0020 0.0016
170 0.0053 0.0059
180 0.0039 0.0042

It should be taken into consideration, however, that for the


temperatures ranging from 140-160°F. the majority of the results,
from which the averages in Table II were t.aken, showed a small
loss in percentage of soluble Ca and Pz05 over that found in the
raw skim milk. For the t,emperatures of 170°F. and 180°F. the
results in every caseshowed a small loss of both compounds in the
fihrate, due to the heating of the skim milk. The difference at
the higher temperatures persists in the results in spite of the fact
that the concentrat,ion of the soluble salts in the serum would
tend to increase, due to evaporation of the moisture in the heating
process.
The conclusions that must be drawn from these results are that
bhe changesfrom soluble to insoluble salts containing calcium and
phosphorus are small and that, even for the higher temperatures
employed, it but slightiy exceeds the possible experimental error.
398 Ca and P Compounds in Milk
In the present study, the average percentage of Ca of samples of
the filt,rat,e from raw skim milk was 0.0272. In terms of CaO, this
is an average of 0.0381 per cent. Since the average CaO content
of the skim milk used was 0.16 per cent, this means that an aver-
age of 23.81 per cent of the CaO content of the skim milk remained
in the filtrate or was in true solution. The lowest analysis showed
0.0206 per cent Ca (0.0298 per cent CaO) and the highest analysis
yielded 0.0333 per cent Ca (0.0539 per cent, CaO).

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


Centrifugal Experiments.
The average gain (+) or loss (-) in CaO content of the filtrate
in terms of percentage was obtained by getting the average loss

TABLE III.
Summaryof Results (Centrifugal Method).
I
Temperature to which I’
heated for 30 min.
Filtrate. Platte Filtrate. Precipi-
tate.
I I
---
“r’. per cent om. per cent om.
Raw (checks) +o.m -0.0126 +1.36 -KOO7 -g8moO75-0.0571
150 to.19 -0.0033 -0.78 -0.0009 -0.0035 -0.0389
160 -0.35 +0.0075 -2.47 +0.0131 -0.0030 +0.0203
170 -1.59 $0.0072, -2.56 $0.0085 -0.0119 +0.0025
180 -4.34 $0.0510 -5.68 +0.0033 -0.0240 +0.1149
190 -'7.58 +0.0903 -6.92 +0.0145 -0.0310 +0.2164
200 -7.04 f0.1003 -8.77 +0.0201 -0.0394 +0.2688
212 -9.75 +0.1246 -9.53 +0.0218 -0.0419 t-O.2954

or gain in gm. of CaO in the filtrates from the raw and heated
skim milk and then calculating what percentage this figure was
of 0.16. The average CaO content of the skim milk analyzed was
0.16 per cent. The average gain (+) or loss (-) in PZ06 in the
filtrate in t*erms of percentage was determined in like manner,
0.2117 being the average percentage of P,Os content of the skim
milk analyzed. TheAe results are summarized in Table III.
The amount of calcium, phosphorus, and ash in the filtrates and
precipitates varied considerably in the different runs. This was
due in part to the speed of the centrifuge. Some runs were made
Raymond W. Bell
at a speed of 38,000 R.P.M., others at 40,000 R.P.X., and also at
speeds between t’hese. To get the R.P.M. of t’hc bowl, a stop-
watch was always used. If it was found that the heated portion
of the skim milk was being centrifuged at a speed less than 40,000
R.P.M., that speed was duplicated as nearly as possible for t,he raw
skim milk from the same source.
Inspection of Table III reveals the fact that with an increase
in t,emperat.ure, t,he amount of soluble CaO and Pz05 in the filtrate
decreases gradually from the amount found in the filtrate from the

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


raw skim milk from t,he same source. The precipitate from these
samples showed a steady gain in the weight of these substances
from the same source. Furthermore, as the temperature of
heating increased, t.he weight of the ash from 100 gm. of t,he filtrate
from t,he heated milk decreased. On t,he other hand, with an
increase in the heat,ing tempcrat.ure, the amount of solids thrown
out by the centrifuge gradually increased over the amount
deposited by the centrifuge from the raw skim milk.

SUMMARY.

Fresh skim milk was heated to various temperatures in order


to study the effect, of heat, on the solubility of the calcium and
phosphorus content. These st,udies were made with the aid of
Pasteur-Chamberland filters and a high speed supercentrifuge.
It appears from the results that there is a loss in t.he soluble
calcium and phosphorus contents of the skim milk due to heat,
and that the amount of the loss depends upon the temperature to
which the milk has been heated.
The result,s from the methods employed indicate that definitely
measurable amount’s of these substances are removed from solu-
tion in milks heated to 170°F. or above.

RIBLIOGRAPHY.

Daniels, A. L., and Laughlin, R., A deficiency in heat-treated milks, J. Biol.


Chem., 1920, xliv, 381.
Flohil, J. T., The electrical conductivity of cow’s milk, Weekhlad, 1911,
viii, 605; Chem. A!utr., 1911, v, 35%.
Grosser, O., cited from Bechhold, H., Colloids in biology and medicine,
translated from the 2nd German edition, wit,h notes and emendations
by Bullowa, J. G. M., New York, 1919, 174.
400 Ca and P Compounds in Milk
Jackson, 1,. C., and Rothera, A. C. H., Milk-its milk sugar, conductivity
and depression of freezing point, Riochem. .J., 1914, viii, 1.
Lane-Claypon, J. E:., Milk and its hygienic relations, New York, 1916.
McCrudden, F. H., The determination of calcium in the presence of mag-
nesium and phosphates: the determination of calcium in uriue, J. Biol.
Chem., 1911-12, x, 187.
Palmer, L. S., The effect of heat on the calcium salts and rennet coagula-
bility of cows’ milk, Proc. Sot. Exp. Biol. rind :lled., 1921-22, xix, 137.
Palmer, 1,. S., and Scott, R. G., The phgsicochemical state of the proteins
in cows’ milk, .J. Biol. (‘hem., 1919, xsxvii, 271.
Rupp, P., Chemical changes produced in cow milk by pasteurization,

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


U. 5’. Dept. Agric., Bureau of Animal Indwstru, Bull. h’o. 166, 1913.
‘,Yan Slyke, L. L., and Boswort!r, A. W., Condition of casein and salts in
milk, New York -4 yric. Exp. Station. T’echn. Bull. 39, 1914.
THE EFFECT OF HEAT ON THE
SOLUBILITY OF THE CALCIUM AND
PHOSPHORUS COMPOUNDS IN MILK
Raymond W. Bell
J. Biol. Chem. 1925, 64:391-400.

Access the most updated version of this article at

Downloaded from http://www.jbc.org/ by guest on September 17, 2016


http://www.jbc.org/content/64/2/391.citation

Alerts:
• When this article is cited
• When a correction for this article is posted

Click here to choose from all of JBC's e-mail


alerts

This article cites 0 references, 0 of which can be


accessed free at
http://www.jbc.org/content/64/2/391.citation.full.ht
ml#ref-list-1