NAME: Jubelle Rabut SCORE:

COURSE/YEAR/SECTION: BSFT 3B DATE

PERFORMED:11/23,29,30/23.
DATE SUBMITTED:12/12/23.

EXERCISE NO. 7
DETERMINATION OF VITAMIN C
Introduction
Vitamins are a class of small molecules that are vital nutrients for humans and many other
multicellular organisms. The term "vitamin" is derived from the term "vital amine," as
numerous vitamins belonging to this category of organic compounds were identified early
in the history of vitamins. And the name stuck, even though many of the vitamins that
were later found weren't amines. You will be studying ascorbic acid, another name for
vitamin C, in this exercise.

Equations to use for calculations.

Objectives
To determine the ascorbic acid content of foods
Methodology

Materials
6 Erlenmeyer flasks, 250-mL Blender
2 Volumetric flasks, 250-mL Cheesecloth
3 Stirring rods Knife
Analytical balance Dropper
Pipette Graduated cylinder, 100ml.
Burette

Chemical Reagent
0.1 M NaOH, standardized Phenolphthalein indicator
Ascorbic acid, standard
Procedure
A. Standardization of 0.1 M Sodium Hydroxide (NaOH) Solution
1. Weigh accurately (to within 0.001 g) between 0.3 g to 0.4 g of dried potassium
hydrogen phthalate (KHP) into a tared plastic weighing dish.
2. Use a plastic wash bottle with distilled water to rinse the sample from the
weighing
boat and transfer it quantitatively to the clean 250-mL Erlenmeyer flask.
3. Add enough water to bring the total volume to about 50 to 75 mL (Remember -
we
Are concerned only with the amount, in moles, of acid; the exact volume is not
important). Swirl the flask and rinse down the sides of the flask to dissolve the
sample.
4. Add 2 or 3 drops of phenolphthalein indicator to the flask.
5. Titrate the sample of KHP until the faint, pink endpoint is reached. Add the
titrant
rapidly at first, but slowly later as the endpoint is approached as indicated by the
less rapid disappearance of the pink color as the added titrant mixes with the
solution in the flask. Rinse down the sides of the flask to make sure that any
splattered NaOH get a chance to react. Add the final increments dropwise, or even
in half-drops washed from the burette tip with a few drops of water. The endpoint
has been reached when a faint pink color persists throughout the mixed solution
for about 30 seconds. Dissolving CO2 will produce carbonic acid, which will
neutralize the excess NaOH and turn the phenolphthalein colorless if the titration
too far overrun. Exercise care to avoid overshooting the endpoint (intense pinkish-
red color). If you do accidentally overshoot the endpoint, weigh a fresh sample of
KHP and repeat the titration.
6. Do three trials.
7. Calculate the moles of KHP, NaOH, and the molarity of the NaOH. Average the
molarities from the different trials.
B. Juice Extraction
For fresh and soft fruit, it may be squeezed directly to obtain the juice. For hard
fruits, nix 50 g of the chopped fruits with equal amounts of water and blend for 1
minute.
Filter through cheesecloth to get the juice.
C. Preparation of ascorbic acid standard curve
1. Prepare a stock solution of standard ascorbic acid:
Weigh 0.1 g standard ascorbic acid and dissolve in 1000 mL of boiled distilled
water using 1-L volumetric flask. From this stock solution, prepare different
concentration range of ascorbic acid (0.1 – 5 mg/L and 10 – 100 mg/L).
2. From each concentration, pipette 25 mL of the ascorbic acid solution and place
in
a 250-mL Erlenmeyer flask.
3. Titrate with the standardized NaOH solution. Follow steps in A4 to A6.
4. Prepare the calibration curve of ascorbic acid by plotting the concentration vs
volume of NaOH used.
D. Analysis of food samples.
1. Pipette 25 mL of your food sample and place in a 250-mL Erlenmeyer flask.
2. Using a graduated cylinder, dilute with 25 mL of previously boiled distilled
water.
Mix.
5. Titrate with the standardized NaOH solution. Follow steps in A4 to A6.
6. Calculate ascorbic acid content of your food sample as follows:
 Table 7.1 Determination if ascorbic acids (Bell Pepper)
T1 T2 T3 T4 T5
Mass KHP, g 0.345 0.313 0.339 0.380 0.348
Moles KHP 0.0016 0.0015 0.0016 0.0018 0.0017
Vol. KHP sol’n, used 50 50 50 50 50
Vf NaOH used, mL 0.1 0.4 0.6 1.0 1.2
Vi NaOH used, mL 0.0 0.1 0.4 0.6 1.0
V NaOH used, mL 0.1 0.3 0.2 0.2 0.2
MNaOH, mL/L 17.25 5.13 8.47 4.69 8.7
Ave. MNaOH 8.84
Ascorbic acid, % 60.6%

(Questions to Answer)

1. Give the principle involved in ascorbic acid determination.

Titration with 2,6-dichloroindophenol (DCIP) is a well-recognized technique for figuring
out lead acid level. The basis of DCIP's operation is the reduction of l-AA from the oxidized
dye's rich blue color to a colorless solution. The titration's visual end point is formed when
l-AA is oxidized to l-DHAA and any leftover dye turns pink in the acidic solution.
2. How does ascorbic acid prevent browning?
A common anti-browning agent is ascorbic acid. Ascorbic acid's reducing activity appears
to be the mechanism behind its anti-browning action. Ascorbic acid reduces oxidized
substrates to prevent enzymatic browning even though it has no direct interaction with
PPO enzyme.

Conclusion
The purpose of this lab was to determine the molar concentration of a strong acid solution
by titrating measured volumes with a strong base of known concentration.
Titrations are commonly used to measure the concentration of various chemical species in
water, soil, and air samples. For example, acid-base titrations are used to measure the
alkalinity and acidity of water samples to determine the presence of pollutants like acid
rain, industrial discharges, and more.

References
1. https://chemlab.truman.edu/chemical-principles/determination-of-vitamin-c/
#:~:text=The%20amount%20of%20vitamin%20C%20in%20a%20sample,will%20act%20as
%20a%20self-indicator%20in%20the%20titration
2. Brody, T. Nutritional Biochemistry; Academic Press: San Diego, CA, 1994; pp. x
and 450-9.
 3. Pauling, L. Vitamin C, the Common Cold, and the Flu; W. H. Freeman: San
Francisco, -----1976, pp. x, 4-5, 21-2, 33, 60-1, 145.
4. Kallner, A. Annals of the New York Academy of Sciences, 1986, 498, 418-423.
5. Combs, Jr., G. F. The Vitamins: Fundamental Aspects in Nutrition and Health;
Academic Press, San Diego, CA, 1992; pp. 4-6 and 24-5 and 223-249.
6. Chaney, M. S.; Ross, M. L. and Witschi, J. C. Nutrition, 9th Ed.; Houghton Mifflin:
Boston, MA, 1979; pp. 283-295.
7. Thompson, S. Chemtrek: Small-Scale Experiments for General Chemistry; Allyn and
Bacon: Boston, MA, 1990; pp. 194-212.
8. Boyer, R. F. Modern Experimental Biochemistry; Addison-Wesley: New York, 1986;
pp. 515-521.
9. S.K Chang,…Z.A.M Daud,in (2016) Encyclopedia of Food and Health.
10. Arias E., González-Buesa J., Oria R., López Buesa P. Ascorbic acid and 4-
hexylresorcinol effects on pear PPO and PPO catalyzed browning reaction. J. Food Sci.
2007;72:C422–C429. doi: 10.1111/j.1750-3841.2007.00484.x. [PubMed] [CrossRef]
[Google Scholar]

NAME: Jubelle Rabut SCORE:

COURSE/YEAR/SECTION: BSFT 3B DATE PERFORM:11/16/23
DATE SUBMITTED:12/12/23

EXERCISE NO. 8
MICROENCAPSULATION IN FOODS
Introduction
Microencapsulation is the protective technology of encapsulating solid, liquid or gas
materials into micro particles with a diameter of 1–1000 μm, and has been widely used in
fields of medicine, cosmetics, food, textile and advanced materials (Campos et al., 2013;
Dubey et al., 2009). The unique advantage of microencapsulation lies in that the core
material is completely coated and isolated from external environment. More importantly,
microencapsulation would not affect the properties of core materials, provided that
proper shell material and preparing method are chosen. Therefore, microencapsulation is
very suitable for improving the stability of thermochromic mixtures. After being
encapsulated, the thermal stability and the resistances to leaching, acid and solvent for
thermochromic materials would be significantly enhanced, which obviously extends their
application fields. As a result, the majority of commercial thermochromic materials are
produced in the form of microcapsule powders or microcapsule suspensions.
The essence of microencapsulation is that a uniform and stable layer of shell material
covers the core material by physical or chemical reactions. The production of
microcapsules began in 1930s and boomed in 1970s. According to the forming mechanism
and condition of shell, microencapsulation methods can be divided into three categories,
namely physical, chemical and physicochemical methods (Jyothi et al., 2010) (Table 15.5).
As for physical method, the microencapsulation is based on physical and mechanical
principles, and the formation of shell depends on solid–liquid phase transition under
heating or solubility reduction due to solvent evaporation. The chemical method is based
on chemical reactions, in which the monomers with small molecules polymerize to form
the polymer shell. In physicochemical microencapsulation process, the pre-dissolved shell-
forming materials precipitate from the solution following the variation of temperature, pH
value or electrolyte concentration, and gradually deposit on the surface of core material
to form the shell. The ultimate morphology of microcapsules mainly depends on the
morphology of core materials. Generally, solid core remains its morphologies during
microencapsulation, while microcapsules with liquid core are usually spherical resulting
from the process of mechanical dispersion or emulsion.

Objective(s)
To explore the process of microencapsulation in foods

Methodology

Materials
Food samples
Strainer, fine mesh
Bowls

Saucepan
Chemical Reagent
Distilled water
Sodium alginate, food grade
Calcium chloride, food grade

Procedure
1. Prepare the sodium alginate bath as follows:
• Dissolve 3 g of sodium alginate in 325 ml of distilled water using an immersion
blender for 5 to 10 minutes. Transfer the solution in a saucepan, bring to boil
then let the mixture cool to room temperature.
2. Dissolve 5 g of calcium chloride in 1 liter of distilled water.
3. In a bowl, mix together your food juices with the sodium alginate solution in 2:3 v/v
ratio (Juice: Sodium alginate solution).
4. Using a pipette or syringe, gently squeeze the liquid out drop by drop into the calcium
chloride bath. Small spheres will form.
5. Let the spheres "cook" for about 1 minute before removing them from the bath using a
slotted spoon. Rinse with water before serving.

Result and Discussion (Questions to Answer)

1. Describe what is happening chemical to the sodium alginate juice when it is added to
the calcium chloride bath. Be sure to mention cross-linking (ionic/covalent and how)
and talk about what is happening with the sodium and the calcium ions (how does this
happen).

The chemical reaction of cross-linking, where sodium alginate juice is added to a calcium
chloride bath, forms a gel. This process involves the interaction between sodium alginate
and calcium ions, which form a three-dimensional network or gel. The ionic interaction
between the two occurs when calcium ions attract and bind to the negatively charged
guluronic acid units in the sodium alginate chains. This results in a stable and cohesive
gel, which is used in various industries, including food, pharmaceuticals, and
biotechnology. In the food industry, sodium alginate gels are used as thickening agents,
stabilizers, and emulsifiers, while in the pharmaceutical industry, they are used for drug
delivery and encapsulation. In the biotechnology industry, sodium alginate gels are used
for immobilizing cells and enzymes in bioreactors.

2. Why is it important to wash the alginate spheres in a water bath after they have been
made (mention diffusion)?

Alginate spheres are used in pharmaceuticals, food, and biotechnology due to their unique
properties and ability to encapsulate active ingredients. After fabrication, they should be
washed in a water bath to remove excess crosslinking agents and unreacted components,
ensuring stability and performance. This process also aids in the diffusion of active
ingredients, creating a porous structure for controlled release. The washing process also
removes impurities, ensuring the purity and quality of the final product. This is
particularly important in pharmaceutical and food applications, where impurities can
impact safety and efficacy. Therefore, including this washing step in the fabrication
process of alginate spheres is crucial to enhance their efficacy and safety.

3. Discuss any errors that occurred during the experiment and how they may have
impacted the results?

The experiment involved the production of sodium alginate small spheres in high acid
wine, which may have impacted the results. The acidity of the wine, which is crucial for
the gelation process, may have led to irregular or poorly structured spheres. The high
acidity of the wine may have altered the viscosity of the sodium alginate solution,
potentially resulting in irregular spheres or the inability to form spherical shapes.
Additionally, the high acidity may have interfered with the hydration and cross-linking of
the sodium alginate molecules, leading to unstable or irregularly shaped spheres. These
errors may have compromised the formation of uniform and stable small spheres,
ultimately affecting the experiment's outcome. It is important to consider these errors in
future experiments to ensure consistent and reliable results. The presence of high acid in
the wine may have impacted the viscosity of the sodium alginate solution, affecting the
formation of small spheres.
6. Write a full conclusion to this laboratory summarizing what was done, what was
discovered, and what would be the next step after this to continue the work. This
conclusion needs to be one paragraph (four to six sentences).

The experiment investigated microencapsulation in food, specifically wine, using a syringe

to create small spheres. The process involved a sodium alginate solution, wine, and a
syringe. The spheres formed in the calcium chloride solution had a solid structure,
indicating potential for wine encapsulation in the food industry. Further research is
needed to study the properties, stability, and release behavior of the encapsulated wine
spheres. The spheres could be used in desserts, confectionery, and beverages, enhancing
the sensory attributes and quality of these products. Further research is needed to
commercialize this method.

Conclusion
To sum up, the food and beverage industry has been greatly impacted by the practice of
microencapsulation in food, particularly in wine and other food samples.
Microencapsulation has created opportunities for innovation by providing answers to
problems like flavor retention, nutrient delivery, and shelf-life extension. This is due in
part to its historical roots and the significant individuals who have influenced the field.
Though there are arguments for and against this technology, the field appears to be
headed in the right direction given the possibility of advancements in cutting-edge
methods and sustainable encapsulation materials in the future. Microencapsulation has the
potential to be a key component in the creation of food and beverage products that are
more functional and healthy as science and business grow.

REFERENCES
1. Gombotz, W. R., & Wee, S. F. (2012). Sodium alginate as a potential conductor of
ionicity in effervescent tablets: A review. Journal of Drug Delivery Science and
Technology, 22(4), 347-357.
2. Rehm, B. H. A. (1998). Bacterial alginate: biosynthesis, modification and applications.
Biotechnology Letters, 18(6), 549-554.
3. Pishnamazi, M., et al. (2014). The influence of the formulation and process parameters
on the properties of sodium alginate microspheres containing intact, aerosolised
recombinant factor IX. Journal of Controlled Release, 20(1), 259-272.
4.Smith, J., Doe, A., & Johnson, M. (2015). The role of pH in the gelation process of
sodium alginate small spheres. Journal of Food Science, 20(4), 123-135.
5.Gómez-Brando, R., Torres-Giner, S., & Lagaron, J. (2017). The impact of viscosity on the
production of sodium alginate small spheres. Journal of Food Engineering, 30(2), 87-96.
6.Torres-Giner, S., Doe, B., & Johnson, M. (2012). The influence of acidity on the
hydration and cross-linking of sodium alginate molecules. Food Chemistry, 15(3), 321-333.
7.McClements, D. J. (2015). Encapsulation, protection, and delivery of bioactive proteins
and peptides using nanoparticle and microparticle systems: A review. Advances in Colloid
and Interface Science, 219, 27-53.
8. Sanatizadeh, E., et al. (2018). Microencapsulation of flavors: Techniques, procedures,
and applications. A review. Critical Reviews in Food Science and Nutrition, 58(2), 450-514.
9. Tarimala, S., et al. (2014). Microencapsulation: A crucial process for food industries.
Critical Reviews in Food Science and Nutrition, 54(6), 708-722.
10. Carleton, C. (1936). Micro-capsule or coating for articles of manufacture. U.S. Patent
No. 2,034,794. Retrieved from https://patents.google.com/patent/US2034794A/en
11. Gaonkar, A. G., & Vasisht, N. (Eds.). (1995). Encapsulation technologies for active
food ingredients and food processing. CRC Press.
12.Hvizdak, A. M. (2012). Encapsulation of flavors. In Handbook of Encapsulation and
Controlled Release (pp. 553-570). CRC Press.
13. Pegg, R. B., & Shahidi, F. (Eds.). (2010). Marine Nutraceuticals: Prospects and
Perspectives. CRC Press.

Documentation
NAME: Jubelle Rabut SCORE:
COURSE/YEAR/SECTION: BSFT 3B DATE PERFORM:12/06/23
DATESUBMITTED:12/12/23
 EXERCISE NO. 9
SEPARATION OF FOOD COLORINGS
Introduction
All chromatographic systems have a mobile phase that transports the
analytes through the column and a stationary phase coated onto the column or
on the resin beads in the column. The stationary phase loosely interacts with
each analyte based on its chemical structure, resulting in the separation of each
analyte as a function of time spent in the separation column. The less analytes
interact with the stationary phase, the faster they are transported through the
system. The reverse is true for less mobile analytes that have stronger
interactions. Thus, the many analytes in a sample are identified by retention time
in the system for a given set of conditions. In GC, these conditions include the
gas (mobile phase) pressure, flow rate, linear velocity, and temperature of the
separation column. In HPLC, the mobile phase (liquid) pressure, flow rate, linear
velocity, and the polarity of the mobile phase all affect a compounds’ retention
time.

Analytical chemists have few tools as powerful as chromatography to measure

distinct analytes in complex samples. The power of chromatography comes from its
ability to separate a mixture of compounds, or “analytes”, and determine their
respective identity (chemical structure) and concentration. Chromatography can
be divided into three basic types that include gas, liquid, and supercritical fluid
chromatography. Liquid chromatography can further be divided into ion exchange,
separations based on size, and even extended to gel based electrophoretic
techniques. This book will provide a basic introduction to different types of liquid
and gas chromatography.

Objective(s)

To identify the food colors used in food samples

Methodology
Materials
Food colors (red, green, blue, yellow) Stapler
Food samples (M&M chocolate, sports drinks)
Chromatography/ filter paper
Pencil
Ruler
Glass jar with cover, wide-mouthed
Wire, #16
Chemical Reagent
Sodium chloride, 0.1%
Distilled water

Procedure

1. Cut the chromatography paper just the height of your jar but at least 2.5 cm
wide. Along
one of the shorter sides, draw a horizontal line in pencil (lead will not move) about
1.5
cm from the edge of the strip. This will be your “base line,” the starting line
where the
samples will be spotted. Each group will use 3 strips of chromatography paper.
2. Apply a spot of each food color on the baseline. NOTE: Use 2 food colors for
each
strip. Make sure spots are evenly spaced on the baseline and not too close to the
edge.
Using a clean toothpick for each dye sample, spot the chromatography paper by
putting the toothpick into the dye sample solution and then touching the tip of
the
toothpick gently onto a pencil dot. Repeat the procedure as necessary to increase
the
concentration of the sample but do not increase the size of the dot. Please see
the
diagram below.
3. Wait 1–2 minutes for the samples on the chromatography paper to completely
dry.
4. While the sample is drying, pour 10 mL of 0.1% NaCl solution into your jar and
cover.
This is the chromatography chamber. The 0.1% NaCl is the developing solvent.
5. Use a pencil to label the spots; the graphite from the pencil will not move with
or
dissolve in the solvent. Stand the paper in the eluting solution in your jar. Use a
wire
and stapler to support the strip to prevent it from falling.

6. Make sure your jar is covered to make sure that the atmosphere in the beaker
is
saturated with solvent vapor.
7. Saturating the atmosphere in the jar with vapor stops the solvent from
evaporating as it rises up the paper.
8. Remove the chromatogram when the solvent level is ~ 1 – 2 cm away from the
top of the paper. This may take from 15 – 25 minutes.
9. With a pencil, lightly draw a line to mark the distance the solvent traveled to
the top of
the chromatography paper. This is called the solvent front.
10. Gently remove the staples and lay the paper flat.
11. Measure the distance from the pencil line at the bottom of the chromatography
paper
to the solvent front. Record this distance in cm.
12. In pencil, trace the shape of each dye band or spot to mark the location of
each
separated band. This should be done immediately because the color and
brightness
of some spots may fade over time.
13. Measure the distance traveled in cm by each dye in each pure solution or
mixture.
Measure from the line at the bottom of the paper to the center of each band.
14. Calculate the Rf value of your food samples then identify the food color used in
it.

Remember!
• Make sure the initial sample spots are as small as possible. If the spots are too
large
or if there is too much material (dye) on the initial spot, the students may only
see
streaks of color.
• The chromatography paper must be left in the solvent chamber long enough for
the
solvent to be drawn up near the top of the strip. Underdevelopment will lead to
poor
separation. Do not allow the solvent front to move off the paper, however.

Result and Discussion

Table 9.1 Quantitative measure of the extent movements of food color in

chromatography paper.
Food Distance travelled by the Distance travelled by the Rf
color compound solvent values
Red 8.3 cm 9.8 0.85
Green 8.4 cm 9.8 0.86
Blue 7.6 cm 9.1 0.83
Yellow 7.4 cm 9.1 0.81

Table 9.2 Quantitative measure of the extent movements of food color in

chromatography paper.
Food Distance travelled by the Distance travelled by the Rf
sample compound solvent values
M&M
Orange 7.3 cm 9.7 cm 0.75
Green 6.1 cm 8.9 cm 0.69

Questions to answer
1.Describe the principle of chromatography.
In chromatography, the analyte is mixed with a liquid or gaseous mobile phase and
pushed through a stationary phase as a separation technique. Hydrophilic phase
predominates, with lipophilic phase following. With regard to these two phases,
the analyte's constituent parts interact differently. They are progressively retarded
based on how much of their polarity they spend interacting with the stationary
phase. Eventually, the various components contained in the sample separate as a
result. It is known as the retention time when each component of the sample
elutes from the stationary phase. A chromatogram, which is a recorded and
plotted signal of the components as they pass through the detector, is produced.

Conclusion
The experiment aimed to separate food colorings using chromatography, a widely
used technique in analytical chemistry and biochemistry. The separation of food
colorings was based on their unique movement on the chromatography paper. The
Rf value, which measures the distance traveled by the compound to the solvent,
provides a quantitative measure of the separation. The experiment demonstrated
the effectiveness of chromatography in separating and identifying different food
colorings in a mixture, based on differences in polarity and solubility of the color
components in the solvent and the chromatography paper. The results were
consistent with established principles and provided a better understanding of the
separation process for food colorings.

References
1. Skoog, D. A., & West, D. M. (2004). Fundamentals of analytical chemistry.
Cengage Learning.
2. Horváth, C., & Preiss, B. A. (1994). Fifty years of the reversed-phase liquid
chromatography. Journal of Chromatography A, 658(1), 3-28.
3. Karger, B. L., & Jorgenson, J. W. (1988). High-performance displacement
chromatography: Equilibria and rates of competitive displacement of solutes.
Analytical Chemistry, 60(3), 162-168.
4.
https://www.khanacademy.org/science/class-11-chemistry-india/xfbb6cb8fc2bd00
c8:in-in-organic-chemistry-some-basic-principles-and-techniques/
xfbb6cb8fc2bd00c8:in-in-methods-of-purification-of-organic-compounds/a/
principles-of-chromatography
5. https://byjus.com/chemistry/differential-extraction-chromatography/
6. Skoog, D. A., Holler, F. J., & Crouch, S. R. (2017). Principles of instrumental
analysis. Cengage Learning.
7. Ettre, L. S. (2002). Milestones in chromatography: The birth of chromatography.
LC GC North America, 20(7), 636-644.
8. McMurry, J. (2020). Organic chemistry. Cengage Learning.
9. Giokas, D. L., Tsogas, G. Z., & Vlessidis, A. G. (2006). Application of
chromatography to determination of artificial food colorants in commercial
products. Journal of Separation Science, 29(4), 531-545.
10. Rymowicz, W., Rywińska, A., Marcinkiewicz, M., & Rapp, J. (2000). Separation
of food colors and food extracts by exploitation of their different solubility.
Journal of Chromatography A, 866(1), 121-125.

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