Effect of agitation on the hydrolysis
of Beta vulgaris L.
When evaluating the agitation, the
percentage of hydrolysis indicated that
there are significant differences
(p <0.05) between the treatments. The
comparison of means was made in which
it was found that between 0, 100 and 150
rpm there is no effect of the treatment;
after 150 rpm, 200 and 250 rpm there are
differences. Due to the fact that there are
two profiles in the agitation, the
inflection point was sought and it was
determined that the agitation of 150 rpm
or less had a better result in obtaining
total sugars (Fig. 4). Chamy et al. (2004)
reported the best agitation speed for the
release of sugars at 400 rpm.
Hydrolysis %
Fig 4. Agitation effect on the hydrolysis of
Beta vulgaris L (▪witness ▪ with acid).
In the tests carried out at shaking speeds
between 250 and 400 rpm (data not
included), no such effect was found and
the energy inversion due to the increase
in agitation did not benefit the overall
hydrolysis process. In the treatments
where the effect of the agitation was
evaluated, an effect was found in the
liberation of sugars on each one of the
controls. However, during the hydrolysis
of the treatments in the that agitation was
evaluated in an acid medium at 0, 100
and 150 rpm, no effects were observed,
for this reason, it was proposed to
evaluate systems with agitation and
without agitation (Fig. 5). In the
experiments carried out with a reaction
time of 0.5 h, there was a difference
between agitated and non-agitated
systems. But with the increase in
reaction time the difference between the
treatments is reduced to almost nil.
Based on this last experiment, you can do
without the agitation and save energy for
this concept. In this experiment (with
and without agitation, Fig. 5), a higher
content of dry mass was quantified,
which increased hydrolysis in all
treatments about 2 times more the initial
content found in the experiments shown
in figure 4, but without affecting the
comparative because it was performed
with the same batch for each test.
Hydrolysis %
Fig 5. Treatments with agitation and without
agitation during the hydrolysis of Beta vulgaris L.
Effect of temperature
With the experiments carried out at
different temperatures (90 ºC and 50 ºC,
in the absence of acid), yields of 18.85
and 8.4% were obtained, respectively.
Between both temperatures there was a
difference of 10.45%, which shows the
effect of temperature on hydrolysis. By
evaluating the combined effect of the
same temperatures and the incorporation
of acid in the reaction, it was observed
that there is an effect on the hydrolysis
gain. In the experiments carried out at 50
° C, in which acid was incorporated in
the reaction medium, the percentage of
hydrolysis was increased from 8.4 to
19.5%, which corresponds to 57% with
respect to the control (Fig. 6).
 Hydrolysis %
Fig 6. Effect of temperature on the
percentage of hydrolysis.
This result was used to determine that 50
ºC would be used as the hydrolysis
temperature suitable for this substrate,
although 3 g / L of total sugars is reduced
in the process, justifying the energy
inversion. Romero et al. (2010) found
during the evaluation of the effect of the
temperature that is at 90 ºC when there is
a better hydrolysis when compared to
that obtained at lower temperatures (60,
70 and 80 ºC). In this case the tendency
is the same, however, handling the
treatments at a lower temperature in
combination with the factors time, mass
and concentration of the acid can result
in better hydrolysis. From the design of
experiments, the effect of the
temperature, the reaction time and the
acid concentration on the amount of
sugars released. With the increase in
temperature from 30 to 40 ºC, the
percentage of hydrolysis went from 22 to
26.5% (Fig. 7).
Hydrolysis %
Fig 7. Hydrolysis of Beta vulgaris L at 30
and 40 ºC (response surface in black and
gray respectively), 150 rpm and 10 g of
mass.
For the tests carried out at 30 ° C, the best
conditions for hydrolysis were
considered: concentration of 0.1 N acid
and reaction time of 4 and 6 h. For the
tests carried out at 40 ºC, the best
hydrolysis conditions were obtained at 6
h and a sulfuric acid concentration of 0.5
N, higher acidic conditions than at 30 ºC
but with a higher hydrolysis yield.
From the design of experiments, the
effect of the temperature, the reaction
time and the concentration of the acid on
the amount of sugars released were
evaluated. With the increase in
temperature from 30 to 40 ºC, the
percentage of hydrolysis went from 22 to
26.5% (Fig. 7). For the tests carried out
at 30 ° C, the best conditions for
hydrolysis were considered:
concentration of 0.1 N acid and reaction
time of 4 and 6 h. For the tests carried out
at 40 ºC, the best hydrolysis conditions
were obtained at 6 h and a sulfuric acid
concentration of 0.5 N, higher acidic
conditions than at 30 ºC but with a higher
hydrolysis yield.
The statistical analysis showed that at 30
° C and reaction time of 2 and 4 h there
is a significant effect of the treatment;
and with 6 h of reaction, the effect is
similar to that of 4 h. The analysis
between groups showed that the acid-
time-temperature interactions have a
greater significant effect in the
treatments carried out at 30ºC over those
generated at 40 and 50ºC (Figs 7 and 8).
It is probable that the above is due to the
short reaction time and the scarce surface
area for the acid attack. In the results
shown in figure 8, the interaction of the
factors (curvature) acid concentration
and reaction time can be appreciated,
however the temperature of 50 ° C had
similar concentration of sugar with
respect to 40 ° C. A minimum of 16
(0.1 0.15 N) and a maximum of 24%
(0.47-0.5 N) is observed. In this
treatment it can be observed that the
parameters of agitation, time,
concentration of the acid and
concentration of the substrate have an
effect on the net yield of the hydrolysis
and can be compared with the individual
treatments the previous experiences to
the establishment of the hydrolysis
conditions (Except to those of figure 5,
where the concentration of the mass was
higher).

Time (h)

Fig 8. Hydrolysis of Beta vulgaris L at

50ºC, 150 rpm and 10 g of mass. The
different levels of sugars released are
shown as curvature of the graph.

The proposed hydrolysis conditions seek

to reduce energy consumption by
agitation and temperature, in addition to
reducing the inhibitory compounds
generated to extreme hydrolysis
conditions, together with the amount of
reagent to neutralize before fermenting.
On the other hand, Beta vulgaris L. is a
suitable candidate to be considered as an
alternative substrate to lignocellulosic
materials such as straw and bagasse,
where its structural complexity hinders
hydrolysis processes and decreases the
yield of simple sugars and therefore the
net yield of ethanol.