pH Measurement
KO Honikel†
r 2014 Elsevier Ltd. All rights reserved.
This article is a revision of the previous edition article by KO Honikel, volume 1,
pp 238–242, © 2004, Elsevier Ltd.
Glossary
pH buffer There exist strong, medium, and weak acids,
and their counterparts strong, medium, and weak bases. The
acids dissociate in aqueous solutions into H+ (proton) and
A– (anion). Strong acids such as HCl dissociate to more
than 99%, medium acids such as lactic acid to 1–5%, and
weak acids to lesser than 0.1%. In water, the undissociated
Introduction
The pH of meat influences its color, water-holding capacity,
flavor, tenderness, and shelf life. Its measurement at different
times postmortem provides information about the forth-
coming quality characteristics. The pH value at the full devel-
opment of rigor mortis is termed as the ultimate pH. To
monitor ultimate pH in a meaningful way, a proper meas-
urement is necessary. However, because of the inherent char-
acteristics of such measurements, it is not always easy to
achieve accuracy and reliability.
What is pH?
The pH of a solution is defined as the negative base 10 loga-
rithm (log10) of the concentration of hydrogen (H+) ions or,
to be more precise, the concentration of the reaction of H+
with a water molecule to produce a hydroxonium ion (H3O+).
For example, a concentration of 10–5 mol l–1 H3O+ in an
aqueous solution has a pH of 5. The pH scale in aqueous
solutions ranges between 0 and 14. Pure water has a pH of 7.0.
In animal bodies most of the organs have similar neutral pH
values, for example, blood has a pH value of approximately
7.3–7.4, muscles are in the pH range of 6.9–7.0.
Hydrogen ions are formed when acids such as lactic acid
(CH3–CHOH–COOH) dissociate in water according to eqn [1].
CH3 –CHOH–COOH þ H2 O⇆CH3 –CHOH–COO– þ H3 Oþ
½1
The dissociation of acids can be more or less complete in
strong acids such as hydrogen chloride, or it can be in equi-
librium in medium strong acids such as lactic acid. Similar to
pH, the degree of dissociation is defined as the pK of an acid
(HA), where K is the dissociation constant of the reaction. The
relationship between pK, concentration of hydrogen ions
(CH+), concentration of anion (CA–), and concentration of
acid (CHA) is shown in eqn [2].
†
Deceased.
262
molecules of medium and weak acids will further dissociate
with increasing pH. With declining pH, more undissociated
molecules will be formed. In this way they provide more or
take up protons and the increase or decrease of pH will slow
down. These compounds buffer the pH change. The
counteractive bases will perform in a similar way but the
other way around.
 
CHþ  CA−
pK ¼ −log ½2
CHA
pK values are found in published tables. For example, for a
0.1 mol l–1 solution at 20–25 °C, the given pK value for lactic
acid is 3.1. Using the pK value, the pH value of a solution of a
weak or medium strong acid can be easily calculated. Using
lactic acid as an example, the pH of a pure 0.1 mol l–1 lactic
acid solution in water can be calculated using eqn [2], where it
can be assumed that the concentration of H+ is equal to the
concentration of A– (anion CH3–CHOH-COO–). Thus CH+ ¼
CA– , substituting the published pK value into eqn [2], results
in eqn [3], which can be solved to give a pH value of 2.04.


3:1 ¼ −log CHþ  CHþ =10–1
8:4  10–4 ¼ ðCHþ Þ2 =10–1
8:4  10–5 ¼ ðCHþ Þ2

1=2
CHþ ¼ 8:4  10–5
CHþ ¼ 9:16  10–3
pH ¼ 2:04 ½3
The pH of an aqueous solution of 0.1 mol l−1 lactic acid is
therefore approximately 2.0.
pH Changes from Muscle to Meat
During the early postmortem changes in muscles of slaugh-
tered animals, the pH declines from approximately 7.0 in the
muscle of a live animal to 5.3–5.8 as its final value, termed as
ultimate pH (pHu). The reason for the pH decline is the for-
mation of approximately 0.1 mol l−1 lactic acid from glycogen
in the anaerobic glycogenolytic pathway.
Buffering Capacity
The discrepancy between the pH of an aqueous 0.1 mol l−1
lactic acid solution of approximately pH 2 (see above) and the
pH of approximately 0.1 mol l−1 lactic acid in meat (5.3–5.8)
is due to the buffering capacity of other constituents in meat
Encyclopedia of Meat Sciences, Volume 1 doi:10.1016/B978-0-12-384731-7.00086-6
 Chemical and Physical Characteristics of Meat | pH Measurement
6.9
6.4 Normal beef
pH Chicken leg
5.9 Normal pork
Turkey breast
PSE pork
5.4
0 2 4 6
Hours postmortem
Figure 1 pH changes of individual muscles of different species. Muscles of different animals and species can differ from those shown here.
that modify the ultimate pH. Buffers are composed of either
moderately strong or weak acids or bases (pK values between 3
and 9) and their soluble salts. These buffering substances in
meat bind H+ ions from the lactic acid formed and lactate
anions prevail. It has been calculated, expressed in formed
lactate units, by the breakdown of glycogen, that the buffering
capacity in meat is between 35 and 50 mmol lactate/pH unit ×
kg of meat in the range of pH 7–5.
In a simple buffer of an acid and its corresponding salt,
eqn [2] also applies. Owing to the addition of the soluble salt
to the acid solution, the concentration of A– is increased. This
means that for the equilibrium to be maintained, CH+ must
become smaller. This means in practice that both lactic acid
and lactate mixtures have a greater pH than lactic acid solu-
tion alone. This happens with meat in a similar way, as ex-
pressed above in ‘lactate terms’ but it is much more
complicated because there are many other constituents in-
volved. Side chains of amino acids, peptides such as carnosine
and anserine, phosphate ions, etc. serve as buffering sub-
stances and lead to ultimate pH values of 5.3–5.8 – more than
3 pH units greater than that of a pure lactic acid solution. In
other words, less than 0.1% of the lactic acid formed is pre-
sent in its dissociated form.
Importance of pH Changes and Ultimate pH in Meat
The ultimate pH of meat is reached at different times post-
mortem depending on species, muscle type, and stress over the
preslaughter period and immediately preslaughter and can
range from 5.3–7.0. The elevation of ultimate pH through
stress affects tenderness and keeping quality. In pork muscles,
a condition arises where the ultimate pH (5.3–5.5) is reached
within 1–2 h postmortem termed pale, soft, exudative (PSE).
In normal pork muscles, pH values of 5.5–5.8 are reached
approximately 6–8 h postmortem. In beef muscles, the ul-
timate pH of 5.5–5.6 occurs at 18–36 h postmortem. In
chicken, ultimate pH values are usually greater, ending at pH
6.0 or more at 2–4 h postmortem under commercial chilling
263
DFD beef
8 12 16 20 24 28
conditions (Figure 1). The rate of pH decline can be reduced
by increasing the rate of carcass chilling; however, the extent of
pH decline is dictated specifically by the glycogen content in
the muscle at the time of exsanguination.
Quite often in bovine muscles, as a result of preslaughter
stress, the pH decline stops at pH values above 6.0 owing to the
lack of glycogen. The reduced glycogen content will result in
less lactic acid. The so-called dark, firm, dry (DFD) beef or dark-
cutting beef is the result (Figure 1). In such cases the pH values
affect many meat properties including color and drip loss.
A low ultimate pH of approximately 5.5 has two effects on
meat. The low pH prevents or retards microbial growth and
the lactic acid/lactate gives the meat a positive flavor com-
ponent. Also, at this pH the aging process results in tender
meat. The rate of pH change postmortem also influences meat
quality. Proteins in muscles before death are ‘native’ due to the
prevailing salt concentration, pH value, and temperature.
Falling pH values at prevailing high temperatures like those in
PSE-prone muscles lead to the denaturation of proteins. These
changes are counterbalanced by the rate of chilling of meat. If
chilling and normal pH decline take place in a controlled way,
as is achieved in many slaughterhouses, then the denaturation
of proteins is limited and there is limited inactivation or ac-
tivation of enzymes (e.g., calpains).
In PSE muscles of pork, the chilling of a carcass cannot
keep pace with the fast pH decline. The final pH value can be
reached at temperatures in the muscle well above 30 °C.
Consequently many proteins denature – the meat becomes
pale due to the denaturation of dissolved sarcoplasmic pro-
teins including the meat-color-producing myoglobin. Also,
proteins in membranes denature and intramuscular fluid can
leak out into the extracellular space. The exudate (also termed
drip loss) increases rapidly and the muscles become soft.
Time of pH Measurement
pH measurements early postmortem, at the end of the
slaughterline at 30–45 min, are very helpful. A pH value of 5.8
264 Chemical and Physical Characteristics of Meat | pH Measurement
and lower at 45 min postmortem indicates that pork muscles
will very likely become PSE meat. An ultimate pH value above
6.0, as in DFD meat, indicates that an early microbial spoilage
of the meat can be anticipated. Thus pH measurements are
important for detecting meat quality characteristics.
Measurement of pH in Meat
Principal Methods
Until the end of the 1950s, the measurement of pH was carried
out using a wide range of coloring substances (pH indicators) in
a solution or bound to paper strips that changed color at
various pH values. Although this was very helpful in an
otherwise colorless solution, in meat or meat extracts, with their
red color, colored solutions could not be used. Paper strips with
pH indicators bound to the paper could be used, but they were
still rather inaccurate with an error of approximately ±0.2 pH
units. Indicator strips with a range of different indicator sub-
stances later enhanced the accuracy to ±0.05 pH units.
Glass electrodes were developed in parallel to metal elec-
trodes in the 1950s. Glass electrodes are based on the principle
that if a glass surface is immersed in a solution, the electrical
potential developed depends on the hydrogen ion concen-
tration. This potential can then be compared with that of an-
other (a reference e.g., a metal) electrode so that, for example,
the pH of a meat slurry can be measured with a good repro-
ducibility. A glass electrode is a proton (H+)-sensitive elec-
trode. The proton sensor is a glass surface (membrane) that
consists of a swollen surface layer (depth 5–500 nm) on both
sides of the glass body of the membrane (Figure 2). On the
outside surface of the membrane, hydrogen ions are replaced
with sodium ions (Na+). On the inside, a buffer solution with
a constant pH is used. An electrical potential is built up, which
is compared with a pH-insensitive reference electrode, usually
a silver–silver chloride or calomel electrode, which is con-
nected with a wick or diaphragm to the solution to be meas-
ured. This means that two electrodes need to be inserted into
the same solution.
As this design was rather complicated, the so-called com-
bination electrode was designed with glass and reference
electrode combined in one shaft. The connection between
the outside (the solution where the pH is measured) and the
reference electrode inside is achieved via a porous ceramic
membrane (Figure 3).
Glass body (0.2−0.5 mm)
Swollen surface layer c. 100 nm
Figure 2 Structure of the glass membrane of a pH electrode.
The early glass electrodes were very fragile and could not be
inserted into a piece of meat. The membrane and the dia-
phragm became rapidly contaminated with fat and protein
particles and had to be cleaned rather frequently. Later a noble
metal wire like platinum was fused in the glass shaft instead of
a porous diaphragm to provide more reproducible results.
Between the outside and inside of the glass membrane,
there is a high resistance of approximately 1010 O. The elec-
trical potential between the inside and outside, measured
through the diaphragm or another reference junction, must
remain undisturbed. Fat or protein particles must not plug the
junction. Furthermore, all parts (measuring solution or meat
(carcass)) must be earthed to avoid false readings. As the glass
membrane exchanges H+ against Na+ ions, pH measurements
are influenced by a number of ions. Measuring in pure (dis-
tilled) water or in a high-salt solution (e.g., in brine with ap-
proximately 5% salt or more or in a salted batter) gives
misreadings. Likewise concentrated protein solutions have a
similar effect. In meat, the misreadings are not significant but
an accuracy below 0.05 pH units is not possible. Temperature
also has a greater influence on readings for the solution
or meat.
In the 1990s, the so-called ion-sensitive field effect tran-
sistor electrodes appeared in the market. With these electrodes,
the surface of a solid-state silicon chip binds H+ ions and
changes its conductivity. However, these surfaces also bind
other charged particles such as proteins and have to be cleaned
very frequently.
Reference
electrode Inner pH
in buffer constant buffer
(e.g. Ag/AgCl)
Glass membrane
Figure 3 A modern combination of glass electrode. The connecting
porous membrane or the noble metal wire is indicated by the black
spot just above the glass membrane.
 Chemical and Physical Characteristics of Meat | pH Measurement
Today, glass electrodes are mechanically robust and
stable enough to be inserted into meat. They measure pH in
meat in a reproducible manner, if calibration and cleaning is
done properly.
Conditions for Measurement
Most chemical reactions are temperature dependent, including
the dissociation of acids and bases. Internationally, pH is
recommended to be measured at ambient temperature of
20–25 °C. However, in meat, temperatures between 0 and
43 °C are common, so this must be kept in mind in inter-
preting pH measurements in meat, as in most cases calibration
is done at approximately 20 °C, yet the meat may be at a
considerably different temperature. Even a temperature ad-
justment at the glass electrode equipment itself corrects only
the temperature dependence of the glass electrode and not the
changes within the meat matrix or meat product, which may
follow a different temperature dependence. Therefore, it is
recommended to accept pH readings of meat to no more than
one decimal place, if the calibration temperature and the
temperature of the meat differ by more than 10 °C. Greater
accuracy demands a calibration at the temperature of the meat.
In these cases the temperature drift of calibration buffers must
also be taken into account (Figure 4).
Calibration
Calibration and associated maintenance are the keys to
meaningful pH measurements. A glass electrode has to be kept
wet during storage. A dry glass electrode has to be conditioned
(hydrated) for approximately 24 h. Electrodes should be kept
in neutral buffer solution or in water. After 30–50 measure-
ments a calibration must be carried out. At least once a day a
calibration must take place.
For calibration, two buffer solutions, usually pH 7.0 and
4.0 or near these values, are taken from the refrigerator (where
they should be stored to prevent microbial growth that affects
the pH value). The solution has to be warmed up to the
ambient temperature of the calibration pH solution. After
6.98
6.96
6.94
pH 6.92
6.9
6.88
6.86
6.84
0 5 10
Figure 4 The effect of temperature on a phosphate buffer.
265
initial cleaning of the combined glass electrode with distilled
water, the electrode is inserted into the standard pH 7 buffer
solution with agitation and the instrument is adjusted, taking
approximately 20 s to reach the pH of the calibration buffer
(e.g., 6.98).
The glass electrode is removed from the first buffer and
cleaned thoroughly with distilled water, then it is inserted into
the second standard pH solution, the pH 4 buffer. After 20 s of
agitation and a stable reading, the pH meter is then set to this
second standard buffer value.
The glass electrode is taken out, cleaned again with water,
and the procedure starting with the pH 7 buffer is repeated
until the values of both calibrating buffers read stable. In many
modern instruments, clear stepwise instructions are displayed
to facilitate calibration.
In most cases two buffers are absolutely necessary. A linear
calibration curve – the linearity between pH 4 and 7 can be
assumed – is defined either by two points (two calibration
buffers) or one point and a constant slope. However, a con-
stant slope recommended sometimes by the manufacturers
cannot be guaranteed with an electrode in regular use as an
electrode ages. This is why calibration with two buffers is
strongly recommended.
Electrode deterioration is noted when the electrode cannot
be calibrated by the process described above. Then it is either
too old or it should be reactivated. To do this, the electrode
surface is first cleaned with warm water (o45 °C) to dissolve
fat films, then the electrode is inserted into a protease solution
(pepsin, chymotrypsin, papain) and left there at room tem-
perature for 1 h. After cleaning with water, the calibration
procedure should be repeated. If the result is not satisfactory,
repeat the protease cleaning step. If successful calibration is
not achieved after carrying out the cleaning process three
times, the electrode should be changed.
Although modern solid-state chip electrodes are robust,
they still need cleaning and the recommendations of the
manufacturer must be followed. Calibration must also be
carried out with these electrodes.
It should be kept in mind that a pH electrode does not have
the same reliability and stability as a calibrated thermometer,
which merely has to be checked once in a while. pH electrodes
15 20 25 30 35
Temperature
266 Chemical and Physical Characteristics of Meat | pH Measurement
must be calibrated regularly as many factors may influence the
ability of the measuring glass surface to exchange H+ against
other cations.
Conclusion
The pH of meat is an important indicator of meat quality
characteristics. It can be used to detect PSE and DFD meat
and it can monitor manufacturing processes. Nowadays, pH
measurements are done with either glass electrodes or solid-
state chip electrodes, but they need frequent calibration. Owing
to temperature differences during measurement, the accuracy of
a measurement is no more than to one decimal place.
See also: Conversion of Muscle to Meat: Color and Texture
Deviations; Glycolysis. Electrical Stimulation. Preslaughter
Handling: Preslaughter Handling
Further Reading
Bendall, J.R., 1979. Relationship between muscle pH and important biochemical
parameters during the post-mortem changes in mammalian. Meat Science 3,
143–157.
Decker, E.A., 2001. The role of histidine-containing compounds on the buffering
capacity of muscle. Proceedings of the 54th Reciprocal Meat Conference, pp.
161−164. Indianapolis, IN: American Meat Science Association.
Honikel, K.O., Fischer, C., 1977. Vergleichende Messung des pH-Wertes mit
Glaselektrode und Spezialindikatorstäbchen bei Rindfleisch. Die Fleischwirtschaft
57 (12), 2175–2177.
Kylä-Puhju, M., Ruusunen, M., Kivikari, R., Puolanne, E., 2004. The buffering
capacity of porcine muscles. Meat Science 67, 587–593.
Maribo, H., Støier, S., Jörgensen, P.F., 1999. Procedure for determination of
glycolytic potential in porcine M. longissimus dorsi. Meat Science 51, 191–193.
Puolanne, E., Kivikari, R., 2000. Determination of the buffering capacity of postrigor
meat. Meat Science 56 (1), 7–13.
Puolanne, E., Reeta Pösö, A., Ruusunen, M.H., et al., 2002. Lactic acid in muscle
and its effects on meat quality. Proceedings of the 55th Annual Reciprocal Meat
Conference, pp. 57−62. Savoy, IL: American Meat Science Association.
Van Laack, R.L.J.M., 2001. Metabolic factors influencing ultimate pH. Proceedings
of the 54th Reciprocal Meat Conference, pp. 158−160. Indianapolis, IN:
American Meat Science Association.
Van Laack, R.L.J.M., Yang, J., Spencer, E., 2001. Determinants of ultimate pH of
pork. Proceedings of the Annual IFT Meeting, New Orleans, LA (Abstract).
Relevant Websites
http://en.wikipedia.org/wiki/PH
http://en.wikipedia.org/wiki/Glass_electrode