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DETERMINATION OF MOLECULAR WEIGHT OF SUCROSE
THROUGH BOILING POINT ELEVATION

Jamorabon, Jasmine Mary A., Labastida Kaye Ann B., Liwag, Danvel C.,
Luteria, Marie Imma B., Malapitan, Dony Lean P.
Department of Biochemistry, Faculty of Pharmacy, University of Santo Tomas
Abstract
Amino acids constitute the primary structure of proteins. Their behavior at different condition generally affects protein function.
Neutralization titration is a common technique used in the laboratory to study amino acid through its acid-base behavior. And to further
understand this concept, this experiment was done. The experiment also aims to plot the titration curve and determine the effect of
formaldehyde to the graph. The analysis was performed first by titrating glycine against a strong base, sodium hydroxide, and that with
the presence of formaldehyde. With the pH values obtained using the calibrated pH meter, a titration curve was plotted. From the data
gathered, the graph of Glycine gives two plateau and an inflection point. It was observe as well that graph of the titration with
formaldehyde is lower than the one without formaldehyde.
based on the result, glycine contains two ionizable group which can
Introduction also be seen in its structure [2]. These are the α -amino and α -
carboxyl group.
Neutralization Titration can be used to monitor progress of reactions On the other hand, it was observed that the graph of the titration with
that produce or consume hydrogen ions [5]. In titration, a solution of formaldehyde is lower than the glycine alone. This is because of the
known concentration is gradually added to another solution of effect of formaldehyde in amino acids. Formaldehyde is composed
unknown concentration until it neutralizes and reaches its molecularly with one carbon and two aldehyde groups. Considering
equivalence point [3]. This analytical technique can be used to the two available oxygen which is partially negative, it will then bond
understand and investigate the properties of amino acids, the to the alpha amino group of the amino acid forming methylol
building blocks of protein. derivative, giving off two methanol side chain. [1] Therefore, the
formaldehyde will mask off the amino group and form alcohol which
Every amino acid has an alpha-carbon chiral center connected to
makes the amino acid more acidic, giving off a lower pH as shown in
four different groups—an amino group, a carboxyl group, a hydrogen
the graph.
atom, and a side chain [4]. Because of this, amino acids are
considered amphoteric substances and react with both acids and An inflexion point was also seen on both graphs. This is the point
bases. Glycine is the simplest amino acid, and is known for its non- where the amino acid glycine behaves as neutral salt and carries no
chirality. This amino acid is essential for many different muscle, net chargev[4], and this point is known as the isoelectric point (pI).
cognitive, and metabolic function [4]. Mathematically, the pI can be determined by taking the average of
the pka values of the buffering region located before and after the
The objectives of this experiment is to be able to construct a curve
said point. One way to obtain the isoelectric point on a titration curve
for glycine titrated with NaOH solution, and analyze the effect of
is to plot the first derivative of the data. The highest peak or the
excess formaldehyde to the titration curve.
maximum slope on the graph of the first derivative represents the
estimated value of the pI. The theoretical pI of glycine is 6.07 while
Methodology the titration curve gave a pI value of 5.09.
Insert your text here

Conclusion
By titrating the neutral amino acid glycine with the standardized
NaOH solution using a calibrated pH meter, the analysts were able
to determine its behavior at different pH. The titration of glycine
revealed two plateaus, which is due to the presence of its two
ionizable group. This means that glycine loses a H+ from acidic
carboxyl group at low pH and loses another H+ from the basic amino
group at high pH. An inflexion point was also observe on the curve
and this was identified as the point at which glycine has a net electric
charge of zero. From other literature, the mechanism of
formaldehyde reaction with amino acid was identified, which is the
formation of the methylol derivative. The effect of this derivative was
observe in the graph as it acidify glycine. These structural changes
of amino acids at different pH could provide a better understanding
of the acid-base properties of protein, since protein plays an integral
Results role in most of the biochemical and physiological event within the
cell.
A prepared NaOH solution was standardized with potassium
hydrogen phthalate and its concentration was calculated to be
0.1943 M. It was used to titrate 0.1 M glycine solution using a References
calibrated pH meter. The pH of glycine was then monitored in every
addition of 2mL of the titrant and the result was plotted below. [1] French, Dexter, and John T. Edsall. "The reactions
of formaldehyde with amino acids and proteins." Advances in protein
15
chemistry 2 (1945): 277-335.
10 [2] Bodner, George M. "Assigning the pKa's of
without polyprotic acids." J. Chem. Educ63.3 (1986): 246.
formaldehyde [3] Acid-Base Titration Curves. (2017). Retrieved from
pH of glycine
5
https://www.khanacademy.org
0 with [4] Voet, D., & Voet, J. G. (2011). Biochemistry.
0 4 8 12 16 20 Formaldehyde Hoboken, NJ: J. Wiley & Sons.
Volume of NaOH
Figure 1. Titration Curve of Glycine with 0.1943 M NaOH with and
without Formaldehyde

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DETERMINATION OF MOLECULAR WEIGHT OF SUCROSE

THROUGH BOILING POINT ELEVATION

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