BKB4493 BIOSEPARATION

ENGINEERING
2013/14-II
CASE STUDY
PROCESS OF PRODUCING THE
BIO-PRODUCTS:CITRIC ACID

GROUP MEMBERS


O RAFIDAH BT MOHAMED ALI
HANAPIAH KE11019
O AHMAD IZZAT BIN ZULKEFLK
KE11036
O OOI CHEE BENG KE11047
O CHONG SOO LING KE11060
O NURUL AIN BINTI IBRAHIM KE12004
Introduction
O one of weak organic acids (a
tricarboxylic acid) widely used as
preservative and conservative in
food.
O additional flavor to increase sour
taste to foods and drinks. Other
uses are for cleaning and chelating
agent, for cosmetics and
pharmaceuticals, as well as
industrial and construction.
O universal intermediate product of
metabolism and its traces are
found in virtually all plants and
animals.


Figure 1.1 shows 3D structure of citric acid. (www.myorganicchemistry.wikispaces.com)
Citric acid can be produced by
fermentation process by
Aspergillus niger, a common
microorganism that is efficient to
produce this acid.
Prior to fermentation process,
citric acid was isolated from
citrus fruits such as ORANGE
and LEMONS where the juices
were treated with calcium
hydroxide to precipitate calcium
citrate, which was isolated and
converted back to the acid using
sulfuric acid (Verhoff, 2005)
Chemical properties
Also known as: Citro, citrate, 77-92-9, Aciletten, Citretten, Anhydrous
citric acid, Chemfill, Hydrocerol A (www.chemicalbook.com)
Molecular formula C6H8O7
Molecular weight 192.12352
Melting point 307.4 F
Specific Gravity/Density: 1.665 g/cm3
pH 1.7 (10% soln)
Density 1.542
Refractive index 1.493~1.509
Storage temp. Store at room temperature
Solubility H
2
O: 1 M at 20 C, clear, colorless
Form grit
Water Solubility 750 g/L (20 C)
Sensitivity Hygroscopic
Stability
Stable. Incompatible with bases, strong
oxidizing agents, reducing agents, metal
nitrate
Physical properties
usually can be found in white crystalline powder
and is described as colorless, odorless crystals with
an acid taste.
denser than water and water soluble.
The pure material is moisture sensitive as it will
undergoes slow hydrolysis.
reacts with oxidizing agents, bases, reducing
agents and metal nitrates. Reactions with metal
nitrates are potentially explosive. Heating to the
point of decomposition causes emission of acrid
smoke and fumes.
Certain measures must be implemented while
handling citric acid in laboratory. Inhalation of dust
irritates nose and throat and contact with eyes
causes irritation. Whilst exposed with fire it will
melts and decomposes. Although, the reaction is
not hazardous
Applications in industry

Citric acid can be found abundantly and its uses are dominantly
in food manufacturing
The largest industrial application of citric acid is for making
detergents.
In liquid detergents, sodium citrate (salts from citric acid) is used
as a builder, to increase the effectiveness of the surfactants, due
to its high solubility and bio-degradability.
While in the form of powdered detergents, sodium citrate is used
as a co-builder and processing aid. Sodium citrate also
contributes alkalinity to enhance surfactant performance. The
environmentally friendly nature of sodium citrate is a major
factor in the use of citrates in the detergent industry.

Applications in industry

Used in plating operations, where it is used as a
chelating agent to control the deposition rate of
metals in both electroplating and electrolysis
plating.
The pharmaceutical industry - as a flavoring and
stabilizing agent in multiple pharmaceutical
preparations. The largest use of citric acid in the
pharmaceutical industry though is for the
effervescent effect it produces when combined
with bicarbonates or carbonates in antacids and
dentifrices
10 m3 of
water per
tonne of
acid
milk of lime containing
calcium oxide (180
250 kg/m3)
120150 minutes.
For 15 minutes
If crystallization is performed at
temperatures below 36.5C, the
citric acid mono-hydrate is
formed and above this transition
temperature citric acid anhydrate
may be obtained.
temperature
below 40C (to
avoid
caramelization),
concentrated
sulphuric
acid (6070 per
cent)
MICROORGANISM
Aspergillus Niger (A. niger) is the microorganism of choice
for citric acid production.

Advantages compared to other microorganism of bacteria,
yeast and fungi:

ease of handling

ability to ferment a variety of cheap raw materials

high yields
(Schuster et al., 2002; Grewal et al., 1995)






NUTRIENT MEDIA
O Carbon source: sweet potato starch hydrolyzate (SPSH) - 153.77
g/L

O Diammonium hydrogen phosphate [(NH
4
)
2
HPO
4
, DAHP] -
3.55 g/L

O Potassium dihydrogen phosphate [KH
2
PO
4
, PDHP] - 2.58
g/L

O MgSO
4
.7H
2
O (0.10 g/L)



(Sankpal et al., 2000)

FERMENTATION PROCEDURE
1. 50 mL of SPSH was measured into 250 mL Duran flasks.

2. Nutrients were added appropriately.

3. The pH of the medium was adjusted using 36.5 g/L HCl and 80 g/L
NaOH buffer solutions.

4. 5% volume fraction of inoculum size was added aseptically to the
flask, which was placed on a clean table for surface fermentation.

5. Fermentation duration and condition - 8 days and pH of 6.






(Betiku et al., 2013)

FILTRATION
O Recovery of citric acid from the surface
process starts with filtration to separate
the mycelium from the culture liquid and
washing of mycelial cake, which may still
contain a significant fraction of the citric
acid produced.
O Filtration of the mycelium from the
submerged process may require the use
of filter aids due to formation of slimy
polysaccharides as by-products

PRECIPITATION
CaOH is
added.
Vacuum
filtration
Dissolve with
dilute H
2
SO
4
.
Filtration
(Pazouki and Panda, 1998)
Objective: To purify the citric acid.
Inlet: Citric Acid and Lime (CaOH)
Outlet: Citric Acid
SOLVENT EXTRACTION
O Amine extraction =method of separation of
carboxylic or hydroxy carboxylic acid from
aqueous solution.
O CA is readily extracted into a number of
organic solvents, such as high-molecular-
weight aliphatic amines (Pazouki and
Panda 1998).
O Wenneresten (1980, 1983) found that
tertiary amines were effective extractants
for CA.

O ADVANTAGE= consuming negligible
amounts of mineral acids and bases and
production of salt by-products.
O Solvent extraction method with tertiary
amines can be further refined for the
technically feasible extraction of CA at pilot
scale
CRYSTALLIZATION
Figure : A forced circulating crystallizer . ( Dyke, 2004)
Objective: To remove the impurities (salt) and ease of storage
Inlet = Citric Acid
Outlet= Citric Acid
EQUIPMENTS
HOLLOW FIBER TUBULAR
MEMBRANE
SPIRAL WOUND MEMBRANE FILTER

O When your ultrafiltration
involve suspended solids,
bacteria, or high
concentrations of
macromolecules, hollow fiber
membranes is used to clarify,
concentrate, and purify
process streams.
O These high-tech membranes
are widely used in submerged
membrane bioreactor
systems, industrial and
municipal water treatment,
and a variety of industrial
applications.
HOLLOW
FIBER
TUBULAR
MEMBRANE
O This design is with a shell and tube
arrangement with polymeric
membranes cast on the inside of
plastic or porous paper components
with lengths from 0.6 - 6.4 m
(Tamime, 2013).
O The feed of the module is passed
through the tubes, accommodating
radial transfer of permeate to the
shell side.
O A single module can consist of 50 to
thousands of hollow fibres. The
diameter of each fibre ranges from
0.6 3 mm with the feed flowing in
the tube and the product permeate
collected radially on the outside
(Munir, 1998).
Hollow Fiber Tubular Membranes
Source : http://www.kochmembrane.com/Learning-Center/Configurations/What-are-Hollow-Fiber-Membranes.aspx
O The advantage of having self-supporting
membranes is easier to clean to its ability
to be back-flushed.
O The disadvantage - Replacement costs
however are high, as one faulty fibre will
require the whole bundle to be replaced.
O Considering the tubes are of small diameter,
using this design also makes the system
prone to blockage.

SPIRAL WOUND MODULE

O The designs of a spiral wound membrane
consist of membrane envelopes (leaves) and
feed spacers which wound around a perforated
central collection tube.

O Feed solution passes axially down the module
across the membrane envelope.

O A portion of the feed solution permeates into the
membrane envelope, where it spirals toward the
center and exits through the collection tube

(Scott et al., 1996).

O - These modules were designed in an effort to pack as
much membrane surface as possible into a given
volume (Senthilmurugan et al., 2005).

O Small scale spiral wound modules consist of a single
membrane leaf wrapped around the collection tube.

O In the large membrane area module, using single
membrane leaf might generate large pressure drop due
to the longer path taken by the permeate to reach the
central collection tube.

O Multiple short leaves have been utilized to keep the
pressure in the module in a manageable level (Van der
Meer and van Dijk, 1997).

Spiral Wound Membrane Module (Scott et al., 1996)
REFERENCES
1. Cheryan, Munir (1998). Ultrafiltration and Microfiltration Handbook. CRC Press.

2. Dyke, S. (2004). Enhanced Forced Circulation: The Continued Solution to High Solids Black Liquor
Crystallization. Il USA: HPD, Plainfield.

3. E. Betiku, O. A. Adesina. (2013). Statistical approach to the optimization of citric acid production using
filamentous fungus Aspergillus niger grown on sweet potato starch hydrolyzate. Biomass and Bioenergy,
55, 350-354.

4. E. Schuster, N. Dunn-Coleman, J. C. Frisvad, P. W. V. Dijck. (2002). On the safety of Aspergillus niger - a
review. Appl. Microbiol. Biotechnol., 59(4-5), 426-435.

5. Frank H. Verhoff (2005), "Citric Acid", Ullmann's Encyclopedia of Industrial Chemistry, Weinheim: Wiley-
VCH

6. H. S. Grewal, K. L. Kalra. (1995). Fungal production of citric acid. Biotechnol. Adv., 13(2), 209-234.

7. Membrane Processing: Dairy and Beverage Applications [Hardcover] by A. Y. Tamime (2013)

8. N. V. Sankpal, A. P. Joshi, B. D. Kulkarni. (2000). Nitrogen-dependent regulation of gluconic and/or citric
acid production by Aspergillus niger. J. Microbiol. Biotechnol., 10(1), 51-55.

9. Pazouki, M., & Panda, T. (1998). Recovery of citric acid - a review. Bioprocess Engineeing 19, 435-439.
O