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Starch Modification
Iris Benardete
S4619714
[email protected]
22 March 2023
Abstract
In this experiment, starch was acetylated with acetic anhydride through an esterification reaction. The
product, acetylated starch, was analyzed by titration and FTIR spectroscopy. The degree of substitution
was calculated as 2.72%, and the FTIR spectrum clearly showed the formation of the carbonyl group. The
results were compared with data from other experimenters and with a fully and non-acetylated starch. A
calibration curve was constructed, and the absorbance values were calculated accordingly.
Introduction
Starch is a white, granular, organic, polymeric carbohydrate with numerous glucose units. Almost all
green plants produce this polysaccharide for energy storage [1]. It is mostly contained in wheat, potato,
corn, rice, etc. Starches can be modified in various ways to increase their stability against excessive heat,
cooling, acid, or time. It can go through physical modifications like cooking, can be treated by enzymes, or
can undergo chemical modification like an acetylation reaction. Acetylation is a chemical reaction where
an acetyl group substitutes a hydrogen atom. This experiment aimed to successfully acetylate a starch
and analyze it by FITR spectroscopy and titration to find the degree of substitution of the acetylated starch
product.
Theory

Figure 1. Acetylation of starch reaction mechanism

Figure 1 displays the reaction mechanism of starch acetylation. First, the most acidic and least sterically
hindered hydroxy group nucleophilically attacks one of the carbonyl groups of the acetic anhydride
molecule. As a result, the nucleophilic oxygen atom gets positively charged, and as a new carbonyl group
forms, an acetate ion leaves. Finally, the base, which was sodium hydroxide in this experiment,
deprotonates the positively charged hydroxy group. As a result, acetylated starch and acetate form.
Titration
A titration is a controlled chemical reaction to find the concentration of an unknown substance. The
titration of a weak acid with a strong base includes the transfer of protons from the weak acid to the
hydroxide ion. [2] In this experiment, titration was used to first determine the unknown concentrations of
the NaOH and HCl solutions that were prepared beforehand and afterward to determine the degree of
substitution of acetate.
Experimental
Day 1 - Acetylation of starch
Suspend starch (20 g) in water (100 mL) and stir until the suspension is homogenous. Measure the pH of
the solution with a calibrated pH meter. Add a few drops of 11M NaOH until the pH reaches 8.5. Slowly
add acetic anhydride (1 mL, 10.5 mmol, 1 eq) simultaneously with 11M NaOH to keep the pH between
8.5 and 9.5. Stir the solution for 10 minutes. Then, decrease the pH to 5.5 by the dropwise addition of
concentrated sulphuric acid. Filter the suspension over a Büchner funnel and wash it with water (4x100
mL). Dry the obtained white powder product in the oven at 80°C overnight.
Day 2 - Analysis of acetylated starch
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Prepare 0.1 M HCl (100 mL) and 0.1 M NaOH (100 mL) solutions. Then prepare two suspensions; a
blank reaction by suspending starch (1.0 g) in water (10 mL) and 0.1 M NaOH (25 mL), and the other one
by suspending acetylated starch (1.0 g) in water (10 mL) and 0.1 M NaOH (25 mL). Stir both reactions for
exactly 2 hours. Determine the exact molarity of the ∼0.1M NaOH solution by titrating a solution of
potassium hydrogen phthalate (300 mg) in water (10 mL) with the ∼0.1M NaOH solution. Then determine
the exact molarity of the ∼0.1M HCl solution by titration of the NaOH solution (5 mL) with a drop of
phenolphthalein. Finally, after two hours of stirring, titrate the blank and acetylated starch samples with
the HCl solution.
Results & Discussion
In this experiment the modification of starch was done by an acetylation reaction. The reaction was run in
basic conditions for the hydroxy group to be deprotonated by the base catalyst and therefore, for the
+¿¿
acetate group to substitute the H proton. Afterwards, multiple titrations were done to determine the
degree of substitution of the acetate group within the starch molecule. The first titration was done to
determine the exact concentration of the NaOH solution. The analyte was 0.3062 grams of KHP dissolved
in 10 mL water while the titrant was the NaOH solution. It took 15.1 mL of NaOH for the KHP solution to
turn pink.
306.2
KHP mols= =1.5 mol=0.0015 mmols
204.22
0.0015
NaOH Concentration= =0.099 M NaOH
0.0151
Then, the HCl solution was titrated with 5 mL NaOH to determine the exact concentration of the ∼0.1M HCl
solution. The NaOH solution went from pink to colorless after 5.3 mL of the HCl solution.
C1V1 = C2V2
(0.099 M)(5 mL) = (5.3 mL) (C2)
C2 = 0.094 M which is the HCl concentration.
Finally, two more titrations were conducted with the blank and acetylated starch samples with the HCl
solution. During the two hours of stirring with NaOH, a hydrolysis reaction occurs to isolate the acetate ion
from the starch molecule in the acetylated sample. The blank solution was also titrated to ensure that the
titration result reflects only the amount of HCl reacting with NaOH and for a controlled set-up. The blank
solution turned colorless after 20.9 mL and the acetylated starch solution turned colorless after 19.1 mL of
the HCl solution. Phenolphthalein was used as the indicator in all titrations. With the obtained results, the
degree of substitution DS AC can be calculated as:
(0.0209−0.0191) x 0.094 x 162
DS AC =¿
(1.0153)−¿ ¿
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Figure 2. FTIR spectra of starches from different data sets and from AVEBE
The FTIR spectrum of non-acetylated starch showed characteristic peaks at 3300 cm -1 and 2900 cm-1,
which correspond to the O-H and C-H stretching vibrations, respectively. The peak at 1020 cm-1 is due to
the C-O stretching vibration in the glycosidic linkage of the starch molecule. In contrast, the FTIR
spectrum of acetylated starch showed characteristic peaks at 1730 cm -1 and 1250 cm-1, which correspond
to the C=O stretching vibration and C-O-C stretching vibration in the acetyl group, respectively. The peak
at 2920 cm-1 is due to the C-H stretching vibration in the methyl group of the acetyl group. The peak at
1020 cm-1 is still present in the acetylated starch spectrum, indicating that the glycosidic linkage is still
present in the molecule. The presence of the acetyl group in the acetylated starch is evident from the
appearance of the characteristic peaks at 1730 cm-1 and 1250 cm-1. These peaks are absent in the non-
acetylated starch spectrum. The peaks in the acetylated starch spectrum are shifted slightly to higher
wavenumbers compared to the non-acetylated starch spectrum. This is likely due to the increased
electronegativity of the acetyl group, which results in a stronger bond and higher vibrational frequency.
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The calibration curve can be made according to the values of absorption and degrees of substitution of
starch 0 and starch AVEBE from the table below:
Absorbance Degree of
substitution

Starch 0 1.01595 0

Starch 85 (AVEBE) 1.02817 85

My data 1.01634 2.72

Student A 1.01656 4.16

Student C 1.01616 1.43

Table 1. Absorbance and degree of substitution values of Figure 3. Calibration curve
starch 0, starch 85 and three different data sets.
The absorbance values of starch 0 and starch 85
were calculated from their IR spectrum. When the
according absorbance and degree of substitution
values were plotted, the calibration curve above is
obtained. The curve gives the equation of
❑
y=6956 x −7067 . The degrees of substitution
of student A and C were calculated with the same
steps that can be seen at the start of the results
and discussion section. When the values are
substituted in the equation, the x value gives the
according absorption value.
The degree of acetylation refers to the number of acetyl groups that are added to the starch molecule.
The degree of acetylation is typically low because the process of acetylation is limited by the availability of
reactive sites on the starch molecule. The extent of acetylation may have been limited by the reaction
conditions such as the type of reagent used, the temperature, and the reaction time. If the reaction
conditions were not optimal, the acetylation reaction may not proceed efficiently, leading to a lower
degree of acetylation. To determine the optimal conditions that give a higher degree of acetylation, more
experiments in different conditions may have been done. Furthermore, excessive acetylation can result in
the formation of a highly cross-linked and insoluble product, which is undesirable in many applications.
Therefore, a low degree of acetylation is often preferred in starch modification experiments to maintain
the desired properties of the starch. The titration results showed that the degree of acetylation of the
experimenter’s modified starch was 2.72%, indicating that the acetylation reaction was actually
successful. This degree of acetylation is within the range commonly used for industrial starch modification
and is expected to have a significant impact on the functional properties of the starch. Student A has
reached a degree of substitution value of 4.16 which was calculated from the chemical analysis, titration
results, and is the highest value amongst the students. This calculation is verified from the IR spectra as
student A’s spectrum shows the highest C=O stretch absorbance value. Overall, the successful
acetylation of starch was confirmed by the FTIR spectrum, which showed the presence of acetyl groups in
the modified starch. The degree of acetylation was significantly higher in the modified starch compared to
the unmodified starch. The modified starch has the potential to have improved physicochemical
properties, making it more useful for various industrial and food applications.

Conclusion
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In conclusion, acetylated starch was synthesized by an esterification reaction with acetic anhydride and
sodium hydroxide as the base catalyst. The product was characterized by titration and FTIR
spectroscopy. The degree of substitution was found to be 2.72% and the according absorbance value
was calculated as 1.01634. The IR spectrum clearly showed the formation of the carbonyl group. The
results were compared with data from other students.
References
[1]“Starch.” Encyclopædia Britannica, Encyclopædia Britannica, Inc., 14 Mar. 2023, https://www.britannica.com/science/starch.
[2] Libretexts. “Titration.” Chemistry LibreTexts, Libretexts, 14 July 2020,
https://chem.libretexts.org/Ancillary_Materials/Demos_Techniques_and_Experiments/General_Lab_Techniques/Titration.
[3] Student manual practicum for Macromolecular Chemistry Practical Starch Modification. March 2023.