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Comparison between the percentage acidity of whole milk and lactose-free

milk through acid-base titration with sodium hydroxide and phenolphthalein as
indicator.

Angela Rodriguez[a], Juan Castañeda[a], Rafael Moreno[a], Juan Romero[a].

Abstract: In the following practice, the percentage of

lactic acid contained in a dilution of whole milk and a
dilution of lactose-free milk is determined in order to
compare the results obtained and establish a difference
between them; the aim is also to relate the percentage of
acidity with the difference between these two types of milk
since although both have this acid in their composition
when decomposing it, they use different methods.

For this practice we used the acid-base titration method

using phenolphthalein as an indicator, since this method
allows us to know the only equivalence point that milk Figure 1: Molecular structure of lactic acid
has, and although this method is quite simple, it allows us
to accurately reach the necessary data to obtain the This is due to the nature of the chemical structure in
expected results. which a carboxyl group (COOH) is present, which favors
the ionization of the hydrogen accompanying this group.
For its part the other hydrogen present in milk is not
potentially ionizable because being a weak acid, it is only
The results obtained are compared with the real value partially deprotonated in solution [2], this causes a less
reported by the food industry, which for milk in good aggressive acidity compared to strong acids; in the case
condition range from 0.12% to 0.16%, si la concentración of milk its capacity is limited by lactic acid, and because of
hallada sobrepasa estos valores, serai indicative que ya this, it has only one ionizable proton.
leche manipulada no cumple la norma y podria estar
generando posibles afectaciones a la salud humana. Another component of milk is lactose (see Figure 2), a
sugar that belongs to the group of chemical compounds
known as carbohydrates, a particularity of this type of
compound is that it can be split into energy-rich
Introduction components [3]. Its presence determines the commercial
nature of milk, which can be whole, and lactose-free,
among others.

Milk is composed of a different number of

macromolecules that contribute to its nutritional value;
however, the composition depends on the animal's diet as
well as the species and type of breed. The components
are: Carbohydrates (lactose), proteins (casein, lacto
albumin), fat (capric, palmitic, oleic acid) [1]. But one of its
main components is lactic acid, which provides flavor,
color and texture. This acid has a weak nature, since it
has only one ionizable proton (see figure 1), which means
that it has only one equivalence point because not all the
protons of its chemical structure are ionized.

Figure 2: Lactose structure

Whole milk is the integral, unaltered, unadulterated, and

unadulterated product without colostrum from hygienic,
regular, complete, and uninterrupted milking of healthy
and well-fed domestic females [4]. On the other hand,
lactose-free milk has added lactase, since it was created

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for those organisms that do not produce this enzyme in Once the calculation has been made, a titration of the
the intestine; by adding this lactase, it specifically splits biphthalate solution is carried out by adding
milk lactose into its component monosaccharides, phenolphthalein. NaOH is added until the solution turns a
glucose and galactose, which are consequently absorbed faint pink color, and the volume value added must be
in the jejunum [5]. taken into account to perform the subsequent calculation
following the equation:
Knowing the above, it is proposed to test the quality of
both lactose-free and whole milk by determining the where: m KC8H5O4 is the amount of solute used in the
degree of acidity of each one in the laboratory; what is solution, n/PM is the molecular weight of the bisphthalate,
expected to be obtained is a lower degree of acidity in CE are the stoichiometric coefficients of the chemical
lactose-free milk, since having less lactose, it is less equation and finally Vt is the volume spent in the NaOH
prone to suffer changes in its acidity due to bacterial titration expressed in liters.
activity compared to whole milk; the presence of this
enzyme in this type of milk can provide more substrate for Titration of both whole and lactose-free milk.
the bacteria to produce a greater amount of lactic acid,
and consequently, it would have a higher percentage of
The acidity of milk is generally determined by volumetric
acidity.
titration analysis using standardized standard solutions of
a base as titrant [8], in this case NaOH, for which the
Experimental Section following chemical reaction must be followed:

General notions: As in the previous section, a dilution of milk will be titrated

using NaOH. This process will be carried out carefully
A technique known as titration was used to obtain the until the dissolution changes color, the data that is
acidity data. Titrations correspond to a group of obtained of NaOH added to the dissolution will be used
techniques that are used for the quantitative for the determination of the acidity taking into account the
determination of multiple measurements. Nowadays, it is following equation:
recognized that the advances and applications of these
techniques have contributed greatly to the development of Where V NaOH is the amount of volume spent in the
the chemical industry and therefore, to the development titration expressed in mL, N is the Normality of NaOH
of chemistry itself [6]. An indicator will be used that expressed in m-eq/mL, A is the correlation constant of
highlights the equivalence point that corresponds to the lactic acid expressed in m-eq, and finally, Vt is the titrated
exact moment where the proton of the substance to be volume of the sample in mL.
titrated is ionized, additionally, as it works at low
concentrations, it does not suppose a noticeable change
Results and Discussion
to the concentration of the solution.

Table 1. Standardization of NaOH for the titration of

lactose-free milk.

Biphthalate mass 0,1031 0,1033

± 0,0001 (g)

Volume of NaOH ± 5,5 5,2

0,05 (mL)

NaOH 0,091 0,077

concentration ±
8,19^-5
Figure 3: Phenolphthalein structure
Average
En la figura 3 se evidencia la naturaleza del indicador a concentration 0,084
usar, a la izquierda de la figura se muestra la forma acida ±0,0001
incolora y la derecha la forma basica color rosa. La
fenolftaleina es un acido debil que pierde cationes H + en
solución.La molécula de fenolftaleina es incolora, en Table 2: NaOH standardization for lactic acid titration in
cambio le anion derivado de la fenolftaleina es de color whole milk.
rosa. Cuando se agrega base, la fenolftaleina (siendo
esta inicialmente incolora) pierde cationes H+formandose
el anion y hacienda que tome coloración rosa. [7]. Biphthalate mass ± 0,0001 (g) 0,1021

Standardization of NaOH for milk titration Volume of NaOH ± 0,05 (mL) 5,25

Concentration NaOH ± 0,0001 (M) 0,095

Initially, the amount of NaOH to be used in the titration
must be known, and the following chemical equation must
be taken into account to calculate the necessary amount
of potassium biphthalate: Table 3: Lactic acid titration data in whole milk.

KC8H5O4 + NaOH NaKC8H4O4 + H2O

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volume dilution with
acidity of each of the samples, the other possible factor
5 mL of milk ± 0,02 25 25 25 as already mentioned may be the very nature of the
(mL) producer, a hypothesis that could be raised is that the
treated milk is not properly "milk" by definition based on
Spent NaOH ± 0,05 the standard, This low level of acidity may be because the
(mL) 0,75 0,7 0,65 "milk" may be diluted with water or may simply have other
types of additives that are associated in some chemical
way with the lactic acid, generating a chemical reaction
Average volume± 0,7 and therefore reducing the presence of lactic acid in the
0,086 (mL) sample, otra causa prar explicar la cercania de los datos
de acidez puede ser fruto de la variabilidad de la
% acidity ± 0,0129 0,1197 estandarización del NaOH, ya que se usaron dos
soluciones diferentes en terminus de concentración, las
cuales difieren en tan solo 0,011 M, este vakor ouede
Table 4: Lactic acid titration data in lactose-free milk. representar un cambio minimo entre los porcentajes de
acidez obtenido puesto que ambos experimentos no se
reealizaron bajo las mismas condiciones . It could be
Volume ± 0,02 (mL) stated that this misreporting is not necessarily serious
25 25 25 since this deviation concerning the range does not
represent any health risk, which could be different if the
Spent NaOH ± 0,05 deviation were higher than the range established by the
(mL) 0,75 0,7 0,65standard. So, what do these below-range values mean?
These results may indicate two things: initially, we have
that the composition of the milk is irregular, specifically
Average volume± 0,7 talking about the presence of lactic acid, The other
0,086 (mL) postulate that can be derived from the analysis is that in
terms of acidity levels, both the whole milk and the alpine
% acidity ± 0,0129 0,1197 lactose-free milk have acidity percentages that are quite
close to each other despite the nature of the milk (see
Figure 4) en esta grafica se puede observer el porcentaje
de acidez de la leche calculado para cada uno de los
volumenes titulados experimentalmente, adiconalmente
se muestra el calculo de la acidez para el promedio de
Figure 4: Comparison of the percentage of acidity volumenes para ambas especies lacteas, with a
between whole milk and lactose-free milk. difference of only 0.0033% of acidity; and although this
difference is minimal, it confirms the initial hypothesis,
which states that a lower percentage of acidity would be
obtained in the lactose-free milk because this type of milk
is less prone to bacterial activity causing changes in its
acidity compared to the lactose-free milk, so it can be
stated that this difference could be a product of the
bacterial activity present in the whole milk.

Conclusions

· The difference between the degree of acidity of

the milk, although minimal, shows that lactose-free
milk is less prone to changes caused by bacterial
activity compared to whole milk.

· The degree of acidity of both types of milk is not

drastically affected despite having different methods
of lactase breakdown, differing by only 0.0033%.
According to the regulations imposed by the Ministry of
Health, the degree of acidity of milk should be between
0.12% and 0.16% [9]. It can be observed that the · The percentages of acidity obtained for both
experimental data are far from this range, specifically the types of milk are below the percentage of acidity
results are located between 0.1164% for lactose-free milk reported by the Ministry of Health, this could occur
and 0.1197% for whole milk, this deviation may be due to due to different natural factors, however, this
multiple factors associated both to the experimental difference does not affect health, since in addition to
practice in the laboratory and to the nature of the milk being very minimal (0.0003% in whole milk and
production plant that was treated (Alpina and Alquería, 0.0036 in lactose-free milk), it is below the reported
respectively). value.

Naturally, as one of the main influencing factors, human · Se puede afirmar que la cantidad de NaOH
error at the time of data collection should be highlighted, gastado en la titulación en directamente proporcional
as well as the possible error in making the calculations of a su concentración, por esta razon al ser la

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concentración de la leche muy baja los moles
agregados reaccionaron mas rapido, al presentarse
en una mayor cantidad, gastando solo 0,77 mL en
promedio en el caso de la leche deslactosada y en el
caso de leche entera 0,7 mL en promedio.

Bibliography

[1] Calderón Z, (2016), Determinación de acidez, p 2.

[2] Atkins J, (2012), Principios de química 5a edición, en:

Ácidos y bases, Editorial Médica Panamericana.

[3] Fabian M, (2009), Determinación de la calidad de la

leche cruda (acidez, densidad, grasa, reductasa, sólidos
totales), aplicando un programa de capacitación en 4
comunidades de la parroquia Pintag, Cantón, Quito. Tesis
ingeniero Zootecnista, p. 31.

[4] Ministerio de salud de España. (1967). Código

alimentario español, Capítulo XV, sección 1a.

[5] Carlos C, (2020) Intolerancia a la lactosa, Editorial

Acta Medica Costarricense.

[6] Morales E, González L, et al. (2019). Técnicas de

titulación ácido-base: consideraciones metrológicas.
Revista Colombiana de química, p.27

[7]. Campos G, Alvarenga E. (2009). Aplicaciones de la

fenolftaleina en algunos campos de la ingenieria civil.
Unidad de investigación y Desarrollo vial. El Salvador.

[8] Chacón A, (2006). Comparación de la titulación de la

acidez de la leche caprina y bovina con hidróxido de
sodio y cal común saturada. Agronomía mesoamericana.
p. 56

[9] Ministerio de Salud de Colombia (1986).

RESOLUCIÓN NÚMERO 02310 DE 1986. Artículo

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