TIT PhD Entrance Exam Presentation

MS Thesis

PRODUCTION OF MICROALGAE IN
PHOTOBIOREACTOR AND CHARACTERIZATION OF
ALGAL FATTY ACIDS FOR BIODIESEL FEEDSTOCK
by

Hoang Anh Hoang

Thesis Advisers
Dr. Joseph Auresenia
Dr. Julius B. Maridable
Prof. Yasunori Tanji

Biofuel August, 2010

 Contents

™Introduction

™Methods, Results and Discussion

™Conclusion and Recommendations

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 Introduction

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 Energy crisis and Global warming

A period of declining non-renewable energy resources, especially petroleum

Æ biofuels especially biodiesel are the best approaches

www.careglobalwarming.com/
http://www.worldculturepictorial.com/blog/blogs/wcpwatchfuleye

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 Vegetable Oil Productivities

40-120
(dry tones/hectare.yr)
Oil productivity

5.08

Vegetable
Oil
2.3
2.0

1.02
0.78 0.82 0.9

Soy Sunflower Peanut Rapeseed Jatropha Coconut Palm oil Algae

Biofuel Source: http://oakhavenpc.org/cultivating_algae.htm 5

 Advantages of Microalgae

20-75% dry weight

composition use carbon dioxide
of neutral lipids
contributing to the
growing cycle Microalgae remediation of
is very fast wastewaters

can grow in produce other co-products or

by-products
seawater or bioreactor

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 STATEMENT OF THE PROBLEM

• What was the composition of the fatty acids

contained in algal strains researched?

+ Fatty acid profiles of many microalgae are not studied yet.

+ The fatty acid profiles vary depending on production process

• What algal strains were suitable as biodiesel feedstock in

terms of Cetane Number (CN) and Iodine Number (IN)?

• What were the optimum conditions for the operation

of photobioreactor system?

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 OBJECTIVES OF THE STUDY

• General Objective
To determine the suitability of some algal strains as
feedstock for biodiesel production based on the fatty
acid profile and to optimize the operating conditions
of photobioreactors and algal strain to obtain the
maximum amount of oil.

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 OBJECTIVES OF THE STUDY
Specific Objectives
• To evaluate the growth of different algal strains in flask
photobioreactor.

• To apply oil extraction and direct transesterification

methods from wet biomass.

• To estimate Cetane Number (CN).

• To evaluate the effect of photoperiod, light intensity,

CO2 flow rate and algal strain on efficiency of the
flask-flat plate photobioreactor system.

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 METHODS, RESULTS AND DISCUSSION

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 PHASES OF THE STUDY

Phase 1
Phase 2
3 algal strains

Flask photobioreactor Flask-flat plate

Media, CO2
photobioreactor

Direct Taguchi optimization

Transesterification

Data analysis

Conclusion

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 METHODS - PHASE 1
Flask photobioreactor
(1:Navicula sp.; 2: Amphora sp.; 3: C.sorokiniana)

Measure COD
1 2 3

Filtration

1 2 3

Direct Transesterification

Biodiesel

GC-FID

Cetane number

Analysis
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 RESULTS – PHASE 1
Flask photobioreactor
(1:Navicula sp.; 2: Amphora sp.; 3: C.sorokiniana)

COD curve, algal growth patterns

1 2 3

Filtration

1 2 3

Direct Transesterification

Biodiesel

GC-FID

FAME profile and CN

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METHODS - PHASE 2 (Taguchi Optimization)

Flask-flat plate photobioreactor

(9 Taguchi runs)

Measure COD
1 2 3

Filtration

1 2 3

Oil extraction

MINITAB

Optimum condition
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 RESULT - PHASE 2
Flask-flat plate photobioreactor
(9 Taguchi runs)

Experimental Duration
1 2 3

Filtration

1 2 3

Oil extraction

Taguchi Analysis

Optimum Condition
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 Flask
Photobioreactors

Schematic Flask Photobioreactor

Working volume (Liter) 3

Light intensity (Lux) 784

Air flow rate (L/min) 9

Photoperiod(L:D, hrs) 24 : 0

Actual Flask Photobioreactor

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 Microalgae and Cultural Media

• Conwy medium (C. sorokiniana) and F/2 medium

(Navicula sp., and Amphora sp.)

• The seawater was obtained in Naic, Philippines.

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 Z
Flask-Flat Plate
Photobioreactors

Schematic Flask-Flat Plate Photobioreactor

Factors Levels

Photoperiod (L:D, hrs) 8:16 12:12 24:0

Light Intensity (lux) 309 590 784

CO2 Flow rate (L/min) 0 0.5 1.0

Algae strain Navicula sp. Amphora sp. C. sorokiniana

Working volume, L 20

Air flow rate, L/min 36

Actual Flask-Flat Plate Photobioreactor

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 Taguchi orthogonal array of L943

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 Algal COD Measurement
algal culture
centrifuge

2.5 ml 2.5 ml
algal culture medium

Catalyst, digestion Catalyst, digestion

solutions solutions

Oven, 150oC, 2 hrs

Cool down, Centrifuge

measuring the absorbance

at 600 nm
Algal COD
Algal solution
= Total COD – Dissolved COD
= Alga + Medium
Total COD Dissolved COD

Algal COD
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Direct transesterification from wet biomass

Wet biomass
(5 grs)

7.0 ml methanol,
0.6 ml sulfuric acid,
4.0 ml chloroform.

Water bath
(90oC for 2 hrs)

Room to 2 ml distilled
water

well mix

Phase separation
(4000 rpm for 15 minutes)

Biodiesel

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FATTY ACID METHYL ESTER PROFILE AND PREDICTED VALUES

Cetane number

CN1 × a1 + CN 2 × a2 + CN 3 × a3 + ... + CN n × an
CN =
100

a1, a2, a3,…,an are the mass fractions.

CN1, CN2, CN3,…,CNn are the cetane numbers for fatty acid components with
known experimental cetane number published

For fatty acids without published experimental CN data,

CN = −14.092 + (6.704 − 2.013db + 0.236db 2 )n − 0.038n 2 (R 2 = 0.947)

n: number of carbon atoms in the original fatty acid,
(Lapuerta, M., J. et al., 2009)
db: number of double bonds

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 Oil Extraction from Wet Biomass

Wet biomass
• 12 ml chloroform (5 grs)
• 12 ml methanol
• 6 ml distilled water
• Glass beads
Ceramic chamber
(blended for 5 minutes)

Centrifuge tube
(Vortex 2 mins + centrifuge)

pre-weighed
graduated glass tube
(60oC, 100oC)

Oil
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 ANALYSIS OF TAGUCHI OPTIMIZATION

• Calculate the output parameter

The oil yield

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 COD STANDARD CURVE
900
COD = 3451.1Absorbance - 299.87
R2 = 0.9963
750

600
COD = 3451.1 (Abs.600) – 299.87
COD (mg/l)

450 R2 = 0.9963
300

150

0
0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4
Absorbance (600nm )

COD Trial 1 Trial 2 Average

0 0.083 0.085 0.084
150 0.138 0.141 0.1395
300 0.176 0.176 0.176
450 0.212 0.208 0.21
600 0.255 0.256 0.2555
750 0.302 0.306 0.304
900 0.35 0.354 0.352

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GROWTH OF MICROALGAE IN FLASK PHOTOBIOREACTOR

2000
1200
1500 1000
C O D (m g /l)

CO D (m g/l)
800
1000 600
400
500
200
0 0
0 1 2 3 4 5 6 0 1 2 3 4 5 6 7

Time (day) Time (day)

Growth Pattern of Navicula sp. Growth Pattern of Amphora sp.

800
700
600
C O D (m g /L )

500
400
300
200 Navicula sp. Amphora sp. C. sorokiniana
100
0
0 1 2 3 4 5 6 7 Algal culture
Time (day) cycle (days) 4 5 5

Growth Pattern of C. sorokiniana

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 FAME profile and Cetane Number

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 FAME profile and Cetane Number
Biodiesel produced from Amphora sp.,
Navicula sp. satisfied all of the standards.

Biodiesel produced from C. sorokiniana met

only the American Standard.

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 TAGUCHI OPTIMIZATION
• Experimental Duration of each Taguchi Run
1500 1000

750

C O D (m g /l)
CO D (m g/l)

1000
500
500 250

0 0
0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8
Day Day

Growth Pattern of Navicula sp. (Run 1) Growth Pattern of Amphora sp. (Run 2)

800
800
600

CO D (m g/L)
600
C O D (m g /L )

400
400

200 200

0 0
0 1 2 3 4 5 6 0 1 2 3 4 5 6 7
Time (day) Time (day)

Growth Pattern of C. sorokiniana (Run 3) Growth Pattern of C. sorokiniana (Run 4)

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 TAGUCHI OPTIMIZATION_Experimental Duration of each Taguchi Run
1500
800
1200
COD (mg/l)

COD (mg/L)
600
900
400
600

300
200

0 0
0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7
Tim e (day) Tim e (day)

Growth Pattern of Navicula sp. (Run 5) Growth Pattern of Amphora sp. (Run 6)
1200 800
CO D (m g/L)

900 600

COD (mg/L)
600
400
300
200
0
0 1 2 3 4 5 6 7 8 9 0
0 1 2 3 4 5 6
Time (day)
Tim e (day)

Growth Pattern of Amphora sp. (Run 7) Growth Pattern of C. sorokiniana (Run 8)

1600

1200
COD (mg/L)

800

400

0
0 1 2 3 4 5 6 7
Tim e (day)

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TAGUCHI OPTIMIZATION_Experimental Duration of each Taguchi Run

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 Input of Taguchi Analysis

where

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 ANALYSIS OF TAGUCHI OPTIMIZATION
• Optimization Based on Oil Yield (mg/L/day)

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 Optimum Condition

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 CONFIRMATION OF THE OPTIMAL TAGUCHI RUN

1600

COD (mg/L) 1200

800

400

0
0 1 2 3 4 5 6 7
Tim e (day)

Growth Pattern of Navicula sp. in the Optimal Taguchi Run

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 Optimum Condition

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 CONCLUSION AND RECOMMENDATIONS

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 CONCLUSION

• The growth patterns of three microalgae in

photobioreactors were evaluated successfully using
COD measurement.

• The main fatty acid components:

+ Amphora sp. and Navicula sp.: palmitic acid (C16:0),
oleic acid (C18:1) and linoleic acid (C18:2).
+ C. sorokiniana: linoleic acid (C18:2) and linolenic
acid(C18:3).

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 CONCLUSION

• Cetane Number (CN) of biodiesel of Amphora sp.,

Navicula sp. and C. sorokiniana were 62.65, 63.61 and
47.02, respectively.

• The best optimum conditions: 24 light hrs (photoperiod),

784 lux (light intensity), 0.5 L/min (CO2 flow rate) and
strain Navicula sp.

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 RECOMMENDATIONS

• It is very important to identify the starting point of the

stationary phase of a specified microalga.

• Explore the use of natural sunlight and larger

bioreactors in the future studies.

• Determine some more biodiesel properties other than

cetane number.

• Apply genetic engineering on algal strains.

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 Acknowledgements
• Faculty of Department of International Development
Engineering, TIT
Prof. Kiyohiko Nakasaki, Dr. Michiharu Abe,…

• AUN/SEED-Net JICA

• Faculty of Chemical Engineering Department, DLSU

• Hanoi University of Science and Technology, Vietnam.

• My family and friends…

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