INTERNATIONAL UNIVERSITY – VNU HCMC

SCHOOL OF BIOTECHNOLOGY
DEPARTMENT OF FOOD TECHNOLOGY

BSc THESIS PROJECT PROPOSAL

Instructor: A.Prof. Pham Van Hung

Presenter: Mai Nguyen Tram Anh
Student’s ID: BTFTIU13006
Pomelo (Citrus maxima) belongs to citrus group and widely planted in South and
Southeast Asia with different varieties.

The total area under pomelo cultivation in Vietnam is 8 170 ha, of which 5 000 ha
produce fruit with annual production of 50 000 tonnes.

Main production areas in Vietnam are Long Vinh, Ben Tre, Dong Nai, Binh Duong, Hue,
Phu Tho, and Ha Tinh provinces and Hanoi
The spongy white peel of pomelo, accounted for up
to 30% of the total fruit weight, is a good source of
pectin (Quoc et al., 2015).

However, pomelo peels are still being considered as

a by-product and there have not been much
research investigate the extraction of pectin from
them
 EXTRACTION AND CHARACTERIZATION
OF PECTIN FROM POMELO (CITRUS
MAXIMA) PEELS USING VARIOUS ORGANIC
ACIDS
Objectives
• To investigate effects of different organic solvents on yield and structure of pectin
from pomelo peels and evaluate the physicochemical characteristics of extracted
pectin.

Research contents
• Extract pectin from pomelo peel using three different organic acids which are
citric acid, acetic acid and lactic acid.
• Determine yield and structure of extracted pectin.
• Characterize physicochemical characteristics of extracted pectin such as degree of
esterification, galacturonic acid content, antioxidant capacity, FTIR
 Sample
Yield
preparation

Degree of
Freeze Drying Drying at 50 C
o
esterification

Extracted with organic Galacturonic acid

acids at pH=3, solid/liquid Blending Analysis content
ratio of 1/30 (w/v)
Antioxidant
Heat at 90 C for
0
capacity
90 min

FTIR
Filtration

Mix with pure ethanol,

ratio 2:1 (v/v)

Centrifugation PROCEDURE
PECTIN CHARACTERIZATION

1. Yield

The yield of pectin extraction was calculated as a function of the pectin mass
obtained from the raw material (dry basis) used, according to following equation:

where, mpectin is the pectin mass obtained and mraw material is the raw material
utilized for extraction
PECTIN CHARACTERIZATION

2. Degree of esterification
Method: based on the methodology proposed by Bochek et al. (2001)

Procedure:

1. Dried pectin (0.05 g) were dissolved in 50 mL of distilled water for 12–15 h

in a drying cabinet at 50 °C in closed flasks.

2. The solution was titrated with 0.05 mol L −1 NaOH until a pH of 8.5±0.2 was
reached. → The used volume of NaOH was recorded as V1

3. The saponification process was carried out by adding 10 mL of 0.5 mol L −1

NaOH for 30 min at 30ºC in a drying cabinet.

4. The solution was then neutralized by the addition of the same volume of 0.5
mol L−1 HCl. The excess of HCl was titrated with 0.05 mol L−1 NaOH → V2
 PECTIN CHARACTERIZATION

3. Galacturonic acid content

Method: based on classical methodology outlined by Blumenkrantz and Asboe-Hansen
(1973)
1. Monohydrated D-galacturonic acid (Sigma, USA) is used as a standard.
2. Adding 1.2 ml of sulfuric acid/tetraborate to 0.2 ml of the sample containing from 0.5 to
20 µg uronic acids.
3. The tubes were refrigerated in crushed ice before shaken in a Vortex mixer and heated in
a water bath at 100°C for 5 min.
4. 20 µL of the m-hydroxydiphenyl reagent was added
5. The tubes were shaken and measured the absorbance at 520 nm by spectrophotometer,
within 5 min.
PECTIN CHARACTERIZATION

4. Evaluation of the antioxidant activity of pectin samples

Method: based on methodology described by Mensor et al. (2001)
1. Six different concentrations (0.250, 0.500, 0.750, 1.0, 1.2 and 1.5 mg mL −1
in 0.1
mol L−1 NaCl) of the extract were used to perform the DPPH assay
2. A 0.075 mmol L−1 DPPH ethanolic solution (1 mL) was added to 2.5 mL of the
sample and the mixture was vortexed at room temperature.
3. After 30 min, the absorbance values were measured at 518 nm
 PECTIN CHARACTERIZATION

5. Fourier transform infrared spectroscopy (FTIR)

The FTIR spectra of pectins were recorded on a Shimadzu, FTIR – 8300

spectrophotometer in the 4,000–500 cm−1 region using potassium bromide (KBr)
pellets (Jiang et al., 2012). The sample was incorporated with KBr and pressed into a
pellet prior to analysis using an infrared spectrometer. The spectrum was recorded
in the transparent mode from 4000 to 400 cm−1, with a resolution of 4 cm.
 STATISTICAL ANALYSIS

All of the analysis tests were done in triplicate except for the antioxidant activity
analysis, which was done in duplicate. The data was analyzed using analysis of
variance (ANOVA) and expressed as mean value and standard deviation,
compared using Tukey’s test at a 5 % confidence level.
PROGRESS AND EXPECTED RESULTS
Content Expected results Time
Part I: Collecting sample Collect at least 3 kg fresh pomelo peel 2 weeks
Part II: Preparing sample 2 weeks
Freeze drying sample Well grind with appropriate particle size
Grinding into powder
Part II: Pectin extraction 1 months
Extract pectin using lactic acid Investigate the convenient method for pectin
Extract pectin using citric acid extraction
Extract pectin using acetic acid
Part III: Characterize extracted pectin 1 months
Characteristic of extracted pectin from different
Characterize physicochemical properties of extraction methods are analyze
extracted pectin
Part IV: Report Analyze obtained data and report the results 1 months
 REFERENCES
Axelos, M., & Thibault, J. (1991). The chemistry of low-methoxyl pectin gelation. The chemistry and technology of pectin, 109-118.
Blumenkrantz, N., & Asboe-Hansen, G. (1973). New method for quantitative determination of uronic acids. Analytical biochemistry, 54(2), 484-489.
Bochek, A., Zabivalova, N., & Petropavlovskii, G. (2001). Determination of the esterification degree of polygalacturonic acid. Russian Journal of Applied Chemistry, 74(5),
796-799.
Jarvis, M. C. (1984). Structure and properties of pectin gels in plant cell walls. Plant, Cell & Environment, 7(3), 153-164.
Jiang, Y., Du, Y., Zhu, X., Xiong, H., Woo, M. W., & Hu, J. (2012). Physicochemical and comparative properties of pectins extracted from Akebia trifoliata var. australis
peel. Carbohydrate Polymers, 87(2), 1663-1669.
Kohn, R. (1982). Binding of toxic cations to pectin, its oligomeric fragments and plant tissues. Carbohydrate Polymers, 2(4), 273-275.
May, C. D. (1990). Industrial pectins: sources, production and applications. Carbohydrate Polymers, 12(1), 79-99.
Mensor, L. L., Menezes, F. S., Leitão, G. G., Reis, A. S., Santos, T. C. d., Coube, C. S., & Leitão, S. G. (2001). Screening of Brazilian plant extracts for antioxidant activity
by the use of DPPH free radical method. Phytotherapy research, 15(2), 127-130.
Mualikrishna, G., & Tharanathan, R. N. (1994). Characterization of pectic polysaccharides from pulse husks. Food chemistry, 50(1), 87-89.
Novosel'skaya, I., Voropaeva, N., Semenova, L., & Rashidova, S. S. (2000). Trends in the science and applications of pectins. Chemistry of Natural Compounds, 36(1), 1-10.
Owens, H. S. (1952). The peptic substances: By ZI Kertesz. Interscience Publishers, Inc., New York, 1951. 640 pp. Price $13.50: Academic Press.
Powell, D., Morris, E., Gidley, M., & Rees, D. (1982). Conformations and interactions of pectins: II. Influence of residue sequence on chain association in calcium pectate
gels. Journal of molecular biology, 155(4), 517-531.
Quoc, L., Huyen, V., Hue, L., Hue, N., Thuan, N., Tam, N., . . . Duy, T. (2015). Extraction of pectin from pomelo (Citrus maxima) peels with the assistance of microwave
and tartaric acid. Int. Food Res. J, 22, 1637-1641.
Sakai, T., Sakamoto, T., Hallaert, J., & Vandamme, E. J. (1993). ⌈ Pectin, Pectinase, and Protopectinase: Production,⌈ Properties, and Applications. Advances in applied
microbiology, 39, 213-294.
Sriamornsak, P. (2003). Chemistry of pectin and its pharmaceutical uses: A review. Silpakorn University International Journal, 3(1-2), 206-228.
Taylor, S., & Walter, R. H. (2012). The chemistry and technology of pectin: Academic Press.
Thakur, B. R., Singh, R. K., Handa, A. K., & Rao, M. (1997). Chemistry and uses of pectin—a review. Critical Reviews in Food Science & Nutrition, 37(1), 47-73.
Venzon, S. S., Canteri, M. H. G., Granato, D., Junior, B. D., Maciel, G. M., Stafussa, A. P., & Haminiuk, C. W. I. (2015). Physicochemical properties of modified citrus
pectins extracted from orange pomace. Journal of food science and technology, 52(7), 4102-4112.
Wilson, L., & Fry, J. (1986). Extensin—a major cell wall glycoprotein. Plant, Cell & Environment, 9(4), 239-260.
THANK YOU VERY MUCH
FOR YOUR ATTENTION