Demonstration Biorefinery for Waste Fish Oil - ccfi.ca Biorefinery for Waste Oil 2013.pdf · Demonstration Biorefinery for Waste Fish Oil Extraction and Processing of Lipids from

Demonstration Biorefinery for Waste Fish Oil Extraction and Processing of Lipids from Marine By-products (Year 1)

Prepared for:

Department of Innovation, Business and Rural Development

Department of Fisheries and Aquaculture

Canadian Centre for Fisheries Innovation

Prepared by:

Centre for Aquaculture and Seafood Development

Fisheries and Marine Institute of Memorial University of Newfoundland

P.O. Box 4920, St. John’s, NL, A1C 5R3

Date: March 27, 2013

Centre for Aquaculture and

Seafood Development

P.O. Box 4920, St. John’s, NL A1C 5R3

Tel: 709-778-0532, Fax: 709-778-0670

Project # -8086 Demonstration Biorefinery for Waste Fish Oil (Year 1)

REPORT COVER PAGE

Project Title:

Demonstration Biorefinery for Waste Fish Oil – Year 1

Project #:

8086

Report Status

√ Applicable Block

Date Progress Report #

Final Report

√

March 27, 2013

Name

Signature

Date

Written by: Deepika Dave

On File March 27, 2013

Written by: Julia Pohling On File March 27, 2013

Reviewed by: Heather Manuel On File March 31, 2013

DEMONSTRATION BIOREFINERY FOR WASTE FISH OIL – YEAR ONE REPORT

Executive Summary This report represents the efforts of the Centre for Aquaculture and Seafood Development (CASD) in

using marine processing waste to produce value-added products (marine oil, biodiesel and animal feed),

establish technology in the area of energy and waste management, enhance the competitiveness of

Newfoundland and Labrador’s seafood and aquaculture industries, and help Canada to meet its

commitment to reducing GHG (green house gas) emissions though building its renewable fuels industry.

It provides a detailed overview over the process developed by CASD in year 1 of the project, which uses

heated oil extraction and chemical transesterification for the production of marine oil and biodiesel

from salmon and seal oil. Biodiesel is a lower alkyl ester of long chain fatty acids and is chiefly made by

transesterification of vegetable oils and animal fats or by esterification of free fatty acids with lower

alcohols.

To date, biodiesel is not readily available in Newfoundland and Labrador, and there are no biodiesel

producers operating within the province. The scope of this project is the development of an

economically viable and environmentally sustainable biodiesel production system for rural

communities in Newfoundland and Labrador and to help marine processing plants to cut down their

operating costs by: (1) diminishing the problem of fish waste disposal; and (2) by providing fuel for the

operation of feed barges, marine vessels and generators located at their remote locations.

Preliminary small scale studies on purchased salmon and seal oil were carried out in terms of pre-

treatment and chemical transesterification. Implementation of a water degumming step on feedstock

oils has reduced the phospholipids content in the feedstock oils. Increasing the alcohol to oil molar

ratio from 6:1 to 12:1 has shown significant improvement in the chemical transesterification reaction

time and glycerol and biodiesel phase separation. Successful recipes from small lab scale experiments

were adopted and transferred to pilot scale biodiesel production. The pilot scale biodiesel plant is

expected to process 220 L of feedstock oil per batch with 98% theoretical yield of biodiesel. Preliminary

results obtained in terms of pilot scale feedstock pretreatment and chemical transesterification are

promising and resulted in 98% (by volume) methyl ester (biodiesel) conversion.

Since the feedstock oils used for biodiesel production is of diverse origin and quality, it was necessary to

implement a standardized test protocol to assess fuel quality and to guarantee engine performance. The

latest official fuel testing standard for B100 biodiesel fuel is ASTM D6751-07b which has been used as a

guideline to develop a simplified test protocol that allows the fuel quality testing to be done onsite with

the use of minimal equipment and in collaboration with Marine Institute.

Purchase and installation of a heat exchanger (Contherm™), a decanter centrifuge for marine oil

extraction and a pre-treatment tank for the pilot-scale biodiesel line have been initiated in year one and

will be completed in year two. Pilot scale marine oil extraction, optimization of the transesterification

process, biodiesel characterization, methanol recovery, development of work instructions and blending

system, environmental analysis of degumming wash water and by product utilization (glycerol and

animal feed) are targeted tasks to perform in year 2.


Contents

1 Introduction ______________________________________________________________ 5

1.1 Practical Importance _________________________________________________________ 5

1.2 Fish oil extraction ____________________________________________________________ 6

1.3 Biodiesel production (Chemical transesterification) ________________________________ 7

1.4 Quality of marine oil and importance of pretreatment ______________________________ 7

1.5 Oil characterization __________________________________________________________ 9

1.6 Factors affecting chemical transesterification ____________________________________ 10

1.7 Separation and Final treatment to FAME (Biodiesel) _______________________________ 12

1.8 Biodiesel storage stability ____________________________________________________ 12

1.9 Fuel properties and specification of biodiesel ____________________________________ 12

1.10 Glycerol refining, management and utilization____________________________________ 17

2 C-ASD approach for Biodiesel Production _____________________________________ 20

2.1 Project scope ______________________________________________________________ 20

2.2 Deliverables for year 1 _______________________________________________________ 20

2.3 Previous challenges _________________________________________________________ 21

2.4 Small scale experiments ______________________________________________________ 21

2.5 Proposed pilot scale fish oil and biodiesel production ______________________________ 23

2.6 Bio-Diesel Logic™ production line ______________________________________________ 30

2.7 Generator installation at Sugarloaf aquaculture site, Bay d’Espoir ____________________ 32

3 Pilot-scale Process Design __________________________________________________ 35

4 Feedstock Oil and Biodiesel Characterization __________________________________ 39

4.1 Proximate analysis of salmon waste streams _____________________________________ 39

4.2 Feedstock oil characterization _________________________________________________ 39

4.3 Biodiesel characterization ____________________________________________________ 41

5 Waste Generation and Management ________________________________________ 46

5.1 Fish oil extraction ___________________________________________________________ 46

5.2 Oil refining and pretreatment _________________________________________________ 46

5.3 Biodiesel production ________________________________________________________ 46


6 Next steps ______________________________________________________________ 49

6.1 Marine oil extraction ________________________________________________________ 49

6.2 Completion of the pilot processing line__________________________________________ 49

6.3 Small scale experiments ______________________________________________________ 49

6.4 Pilot scale experiments ______________________________________________________ 49

6.5 Biodiesel characterization ____________________________________________________ 50

6.6 Development of work instructions _____________________________________________ 50

6.7 Development of a blending system _____________________________________________ 50

6.8 Glycerol management _______________________________________________________ 50

6.9 Environmental analysis ______________________________________________________ 50

7 References ______________________________________________________________ 51


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1 Introduction

1.1 Practical Importance Canada’s commercial fishing industry is valued at approximately $2 billion a year with its aquaculture

industry worth $925 million. Of the 850,533 tons marine landings and 163, 063 tons aquaculture in

2011, 34% and 10%, respectively came from Newfoundland and Labrador (DFO, 2013). There are 187

registered fish processing facilities in Newfoundland, ranging in size from feeder plants (processing fish

to the fillet) to large year-round plants (processing fish into various fresh and frozen products including

secondary processing) (DFA, 2013). In 2011, Salmonid production accounted for 14,264 tons (93%) and

was valued at $112 million. Of the 144 licensed aquaculture plants, 81 produce salmonids (DFO, 2013).

Processing of fish generates large amounts of solid wastes, up to 30-80% of the body weight of the

processed fish. Currently, most of the fish processing waste is dumped at sea or in landfills (Murugesan

et al., 2009).

Sealing has thrived in Newfoundland and Labrador for hundreds of years. The industry grew, bringing a

great deal of foreign investment and the growth of secondary employment in such trades as ship

building, carpentry, and refining. There are approximately 12,500 licensed sealers in the province. While

the seal market price varies from year to year, the sealing industry has been worth over $55 million per

year to the province’s economy (DFA, 2013).The current population of 7.66 million seals consumes

about 1.78 million tons of fish per year compared to total fish landing of 0.96 million tons per year (DFO,

2013). Only relatively unrefined seal products (raw seal skins, seal oil and seal meat) can be identified in

Canada’s export statistics (DFO, 2006). While the focus is directed on the method used for killing seal,

much of the negativity arises from the utter waste of the resource and environmental impact of

discarding seal carcasses into the ocean.

Thus, it can be advantageous to develop by-product applications that demand large volumes of seal

carcass and fish waste, thereby making these industries viable and more environmentally friendly.

The continuously increasing demand for energy has been translated into increased cost of crude oils,

shortage of fossil fuels and intensified emission of greenhouse gases worldwide. If the utilization of fossil

fuels is continued at the present rate, local air quality will deteriorate severely and global warming will

increase beyond repairable extent (Fukuda et al., 2001; Akoh et al., 2007). Renewable energy resources

of biological origin (biofuels) have smaller net greenhouse gas emissions. Currently, biodiesel and

bioethanol production are gaining momentum all across the globe due to the shrinking supply of oil

reserves, security of source, cost of production and the impending threat of global warming (Demirbas,

2007). However, sustainable production of biofuels will require a resourceful biomass conversion

process.

Biodiesel is a biofuel that is obtained from plant and marine oils or animal fats. Biodiesel, as a diesel-

equivalent, has a potential share among biofuels of about three quarter of all refinery distillate fuel oils.

In comparison to petroleum diesel, biodiesel significantly reduces emissions of carbon dioxide (about

50-60%), sulfur dioxide and harmful air pollutants, in particular asthma-causing soots. GHG emissions


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can be reduced by 10-20% and 40-90% with the use of at least 20% (B20) and 100% (B100) biodiesel

blends, respectively (Hinton et al., 1999; Srivastava and Prasad, 2000; Subramanian et al., 2005).

One of the biggest challenges in biodiesel production is the availability of feed-stock. There is a concern

about using plant derived oils and fats since the crops used for biodiesel production are also needed for

food, feed and oleochemical industries. Biodiesel factories must compete with food, cosmetic, chemical

and livestock feed demands. There is also an environmental concern because an increased demand for

vegetable oils requires an increase in the use of fertilizers which contribute to greenhouse gas

emissions. In fact, biodiesel production from heavily fertilized crops could result in a 70% increase (from

the current value) in greenhouse gas emissions (McNeff et al., 2008; Jegannathan et al., 2008;

Ranganathan et al., 2008).

These factors have necessitated the need for the development of a bio-refinery approach for the

production of biodiesel from waste based and cheap biomass rich in oil such as seal fat and fish oils. It

is anticipated that this approach will result in sustainable biofuels, seal and fishery industries.

To date, biodiesel is not readily available in Newfoundland and Labrador, and there are no biodiesel

producers operating within the province. Newfoundland and Labrador’s seafood industry generates

average 102,850 tons (25% of Canada’s fish waste) of processing discards from which valuable oils can

be recovered. Newfoundland and Labrador has the potential to produce 2600 tons of marine oils

extracted from seal harvesting and fish processing waste as the largest potential source of biodiesel

feedstock that can be converted into approximately 2,548 tons of biodiesel (assuming a 98% yield)

(Manuel et al., 2006). Specific to Salmonid industry waste, 360 tons of fish oil can be extracted and

utilized to produce biodiesel. Converting marine oil into biodiesel would benefit the marine industry

sectors in reducing the disposal cost of theses waste to landfills and utilize biodiesel for operating feed

barges, marine vessels and generators located at their remote locations. After the fish oil has been

extracted, residual biomass can be used as a feed stock for production of biomethane and bioethanol via

fermentation or fertilizer or feed for animals.

The long-term objective of this project is the development of an economically viable and

environmentally sustainable biodiesel production system for rural communities in Newfoundland and

Labrador. An enormous capitalization potential exists for the sources of marine processing wastes for

the production of fish oil, biodiesel and fish meal. However, characterization of fish processing wastes

from different plants and development of proper handling, pretreatment and extraction procedures are

necessary for the overall process development for fish oil and biodiesel.

1.2 Fish oil extraction

Fish oil can be extracted by various methods such as physical and thermal processes, bio-chemical

processes (fermentation, digestion, ensilaging and hydrolysis) (Peterson et al., 1983; Faaij, 2006) and

supercritical fluid extraction (Manirakiza and. Covaci, 2001). Physical and thermal processes (heat

extraction) have been widely commercialized and proven their adaptability due to high yields of oil. In


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the thermal process, fat tissues are heated until fat cells burst and release the oil into solution from

where it can be further purified by a centrifugation step.

1.3 Biodiesel production (Chemical transesterification) Transesterification is the process of reaction of a fat or oil (triacylglycerol-TAG) with an alcohol to yield

biodiesel (fatty acid methyl ester-FAME) and glycerol in the presence of a catalyst (chemical-alkali or

acids; biological–enzymes or microorganisms) (Akoh et al., 2007; Ranganathan et al., 2008). It has been

shown that the transesterification seems to be the best practical option (compared to dilution, micro-

emulsification or pyrolysis) since this process can significantly reduce the high viscosity of oils and

produces a biodiesel with the same physical properties as petroleum diesel fuel (Ma and Hanna, 1999).

Either KOH or NaOH are used as an alkali catalyst together with an alcohol (methanol or ethanol) to

convert oil into biodiesel. The schematic principle of the transesterification reaction is shown in Figure 1.

The simplified form of its chemical reaction is presented in Equation 1. Where R1, R2, R3 are long-chain

hydrocarbons, sometimes called fatty acid chains. Normally, there are five main types of chains in

vegetable oils and animal oils: palmitic, stearic, oleic, linoleic, and linolenic. When the triglyceride is

converted stepwise to diglyceride, monoglyceride, and finally to glycerol, 1 mol of fatty ester is liberated

at each step (Ma and Hanna, 1999).

(1)

1.4 Quality of marine oil and importance of pretreatment The properties of marine oils are less uniform compared to fresh vegetable oils because of the physical

and chemical changes mainly due to oxidative and hydrolytic reactions that take place during handling,

stabilization, storage and the oil extraction process. High quality crude oils may be obtained by proper

handling of raw material such as minimizing damage to fish and proper chilling after landing. Following

fish oil extraction, impurities need to be identified and treated before proceeding to the fish oil derived

biodiesel production.

1.4.1 Phospholipids removal (Degumming)

Literature states that fish oil contains higher amounts of phospholipids than vegetable oils. These

“gums” have an emulsifying effect and inhibit the separation of soaps and lower the yield of neutral oil.


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Figure 1: Schematic of the transesterification reaction

Phospholipids can be removed in a pre-treatment step (Degumming) by adding 1-3% water and stirring

for 60 min and then separating the water layer by centrifugation or settling (Wanasundara et al., 1998).

1.4.2 Acid pretreatment

It has been reported that salmon fish oil contains higher amounts of free fatty acids (FFA) along with

moisture which are required to be removed to prevent soap formation as FFA of salmon fish oil are

sensitive to alkali catalyst and react to neutralize the FFA in the oil as shown in Equation 2 (Vicente et al.,

2004). The acid value of the oil should be less than 1 mgKOH/g oil, the amount of FFA should be in the

range of 0.5-2.5 wt% and the feed oil should be anhydrous (moisture content less than 0.3%) (Freedman

et al., 1984; Meher et al., 2006).

(2)

1.4.3 Oxidative stability

Lipid oxidation is a major cause of deterioration and spoilage for fish oil and biodiesel. The fundamental

substrate involved in lipid oxidation reactions is unsaturated fatty acids. Different oxidation processes

take place including: auto-oxidation, photo-oxidation and thermal oxidation (Frankel, 1991).

Characteristic changes associated with oxidative deterioration of marine oils include the development of

unpleasant smells and odors as well as changes in color, viscosity, specific gravity and solubility

(Wanasundara et al., 1998).

Free fatty acid Catalyst Soap Water


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1.5 Oil characterization It is essential to analyze extracted oil after refining and before the transesterification process to produce

biodiesel. While some procedures require extensive preparation and high investments for equipment

and a laboratory facility, some tests are quick and easy and give a good indication of the quality of the

oil. Feedstock for biodiesel production are given priority selection and pre-treatment according to the

level of free fatty acids, impurities, moisture content, unsaponifiable matter, p-anisidine value, and

peroxide value (St. Angelo, 1996; Wrolstad et al.,2005; Boran et al., 2006; O’Brien, 2009). The

procedures identified for fast testing on site are incorporated with consideration of reliability, easiness

and equipment such that conducting the test is easy and reliable and can be performed with a minimum

of equipment.

1.5.1 Free Fatty Acid content

The acid number determines the amount of free fatty acids in an oil sample. A high FFA content can lead

to increased soap formation during transesterification which decreases the yield and prevents phase

separation. Furthermore, FFA reacts with the catalyst so that the amount of catalyst added to a

transesterification reaction is adjusted depending on the FFA content. The free fatty acid content is

determined with a simple, colorimetric acid-base titration.

1.5.2 Visual appearance and smell

Visual appearance and smell is not an official test, however it gives good idea about the quality of the

oil. Oil is examined visually for residues that collect at the bottom of the barrel, particles, cloudiness,

bubbles or streaks. Then the oil is stirred and observed for streaks, color changes, cloudiness or

particles. A rancid smell of the oil is an indication of oxidation.

1.5.3 Water content

The water content in oil must be low to prevent excessive soap formation during the transesterification

reaction. Oil can contain up to 1500 ppm of water without appearing turbid, depending on the feedstock

characteristics. If the water content is above 1500 ppm, a drying step is necessary.

1.5.4 Peroxide value

The peroxide value is the measurement of the primary oxidation product hydrogen peroxide. The

oxidative process of oils and fats is one of the main causes of the deterioration of the principal

organoleptic and nutritional characteristics of foodstuffs. The most widely used chemical test for the

determination of fats and oil quality is the peroxide value test. The number of peroxides present in the

oil is an index of their primary oxidative level and its tendency to go rancid. The complex oxidation

process can be summarized into two phases: in the first phase, fatty acids react with oxygen and

determine odorless compounds as peroxides; during the second phase the peroxides degrade into many

substances such as volatile aldehydes, responsible for the rancid odor and flavor, and in a non-volatile

portion. The lower the peroxide value the better the oil quality and its state of preservation. The primary

oxidation products are normally measured with the Peroxide Value (PV) test and the secondary products

with the p-Anisidine test.


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1.5.5 P- Anisidine value

The p-anisidine value is used to measure the secondary product of oxidation and determines the

aldehyde in the lipid, primarily 2-alkene present in the fat. Aldehyde present in the oil and the p-

anisidine reagent react under acidic condition. The color obtained not only depends on the aldehyde

present, but also their structure. Further degradation of lipids generates off-flavors and off-odors. On

the contrary, other tests consider the volatile portion of aldehydes and, due to their intrinsic variable

nature, result in less reliable data.

1.5.6 Fatty acid composition

The fatty acid composition is very important as it determines the stability, viscosity and reactivity of the

oil. Furthermore, the transesterification reaction is influenced by those characteristics, as well as the

performance of the final product biodiesel. For example, very long fatty acid chains will produce

biodiesel with a high cloud point and results in poor performance in cold weather conditions. High free

fatty acid content require oil pre-treatment before chemical transesterification and high amounts of

unsaturated fatty acids require stabilization of the oil and biodiesel in the early stage of the biodiesel

process. Also the amount of phospholipids will determine the necessity of a degumming step.

While a full fatty acid analysis is not required for each batch of oil, this analysis is an integral part of the

process development and should be performed every couple of batches or when problems are

encountered to monitor potential changes in feedstock quality.

1.6 Factors affecting chemical transesterification There are several factors which affect the rate at which transesterification proceeds and the ultimate

yield of biodiesel. These include: (a) type of alcohol, (b) alcohol to oil molar ratio, (c) reaction

temperature and time, (d) catalyst type and concentration, (e) water activity and (f) mixing intensity.

Therefore, a detailed study on alkali based transesterification is very important for the fish processing

wastes available in Newfoundland and Labrador.

1.6.1 Type of alcohol

The most frequently used alcohol (acyl-acceptors) that can be used for transesterification include:

methanol, ethanol, propanol, butanol, amyl alcohol, octanol, and branched alcohols (Fukuda et al.,

2001). Methanol is most widely used for FAME production, and the reaction is known as methanolysis.

In addition, ethanol is also used but it is relatively expensive, less volatile and less reactive, renewable

and eco-friendly as it is produced from agricultural products when compared to methanol (Bacovsky et

al., 2007).

1.6.2 Alcohol to oil molar ratio

The optimum molar alcohol to oil ratio is generally based on the reaction system, feedstock, catalyst and

type of alcohol used. Theoretically, for each molecule of triglyceride, three molecules of alcohol are

needed to produce three molecules of FAME. Alcohol is used in excess to shift the equilibrium towards

the formation of esters (Robles-Medina et al., 2009). A molar ratio of 6:1 is normally used in industrial

processes because the yield of FAME is higher than 98% by weight. The molar ratio has no effect on acid,


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peroxide, saponification and iodine value of methyl esters. Depending on the feedstock oil, optimal

molar ratios of methanol to oil were reported between 3:1 and 24:1 (Matassoli et al., 2009).

1.6.3 Reaction temperature and time

Literature revealed that reaction temperature and time has a significant effect on the rate of

transesterification. However, the reaction can be conducted at room temperature if sufficient time is

provided (Srivastava and Prasad, 2000). However, a longer reaction time does not necessarily increase

the conversion but favors the backward reaction (hydrolysis of esters) which results in a reduction of

product yield (Leung and Guo, 2006). It is already established that in most cases, the reaction

temperature is kept close to the boiling point of methanol (60–70°C) at atmospheric pressure for a given

time. Such mild reaction conditions require pre-treatment (degumming or esterification) of oil for the

removal of free phospholipids and fatty acids. Simultaneous esterification and transesterification can

take place under high pressure (9000 kPa) and high temperature (240°C) so a pretreatment step can be

eliminated if the reaction is carried out under these conditions (Hwi, 2006).

1.6.4 Catalyst type and concentration

Alkali, acid, enzyme or heterogeneous catalysts are used in the transesterification reaction, among

which alkali catalysts like NaOH and KOH are more effective and used industrially due to their

availability, low cost and large continuous-flow production processes. Advantages of using an alkali

catalyst are: (a) their ability to neutralize free fatty acids and water to mediate the reaction (b) their

higher yield (c) a reduced reaction time (d) and, a lower concentration of alcohol needed for the

reaction to continue to completion (Komers et al., 2001). While using NaOH as a catalyst, it is very

essential to maintain its anhydrous form and store it in a dry place as presence of moisture leads to the

production of soap during transesterification. Concentrations ranging from 0.5 to 1.0% (w/w) have been

found to yield conversion rates of 94 to 99% (Saka and Dadan, 2001).

1.6.5 Water activity

The water/moisture content of the reactants play a vital role in the transesterification reaction as a

water content above 1500 ppm in oil significantly increases soap formation during transesterification.

This produced soap also increases the viscosity of the reaction mixture, sometimes causing gel

formation, thereby making the separation of glycerol from ester difficult (Wright, 1944).

1.6.6 Mixing intensity

Mixing is very important in the transesterification reaction, as oils or fats are immiscible with NaOH–

MeOH solution. Once the two phases are mixed and the reaction is started, stirring is no longer needed.

Methanolysis can be conducted with different rates of stirring ranging from 180-600 rpm. More gentle

mixing methods should be implemented to prevent formation of tiny glycerol bubbles (emulsion).

Reaction time is an important factor for determining the yield of methyl esters (Ma and Hanna, 1999).


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1.7 Separation and Final treatment to FAME (Biodiesel) An important step in the production of biodiesel is the final treatment of the raw fatty acid alkyl ester.

Once the transesterification reaction is completed, two major products exist: esters (biodiesel) and

glycerol. Due to the higher density of glycerol compared to the biodiesel phase it settles at the bottom

of the reaction vessel, allowing an easy separation from the biodiesel phase. Phase separation can be

observed within 10 min and can be completed within several hours of settling. The reaction mixture is

allowed to settle in the reaction vessel in order to allow the initial separation of biodiesel and glycerol,

or the mixture is pumped into a settling vessel. In some cases, a centrifuge may be used to separate the

two phases (Van Gerpen et al., 2004).

After transesterification, the obtained biodiesel and glycerol are contaminated with unreacted catalyst,

alcohol, oil and soap. Although the glycerol phase tends to contain a higher percentage of contaminants

than the biodiesel, a significant amount of contaminants is also present in the biodiesel. Therefore,

crude biodiesel needs to be purified before use (Schumacher, 2007).

Washing of biodiesel is essential to remove remnants of the catalyst, soaps, salts, residual alcohol, and

free glycerol from the crude biodiesel. Normally, neutralization and alcohol stripping are the steps

offered to crude biodiesel before the washing step. Unreacted alcohol should be removed with

distillation equipment before the washing step to prevent excess alcohol from entering the wastewater

stream (Van Gerpen et al., 2004). Generally, three main approaches are adopted for purifying biodiesel:

water washing, dry washing (Cooke et al., 2005), and membrane extraction (Gabelman and Hwang,

1999; He et al., 2005).

1.8 Biodiesel storage stability Biodiesel is safe to store and the properties of biodiesel should remain stable even after long term

storage. There are several key factors that need to be considered for the storage of biodiesel, including

exposure temperature, oxidative stability, fuel solvency, and material compatibility (Mudge et al., 1999).

Viscosity, peroxide value and more dramatically, Rancimat Induction Period have shown changes

compared to other properties of biodiesel during storage (Meher et al., 2006). Most pure biodiesel is

generally stored at 7 and 10°C to avoid the formation of crystals which can plug fuel lines and fuel filters.

An antioxidant additive is necessary to maintain the acid value and fuel viscosity and avoid formation of

gums and sediments. Additionally, biocides need to be added for prevention of biological growth in the

fuel. Biodiesel storage tanks made of aluminum, steel, Teflon®, and fluorinated polyethylene or

polypropylene should be selected (Mittelbach and Gangl, 2001).

1.9 Fuel properties and specification of biodiesel Since biodiesel is produced from feedstock of diverse origin and quality, it was necessary to install a

standardization of fuel quality to guarantee engine performance without any difficulties. The

parameters, which define the quality of biodiesel, can be divided into two groups. One group contains

general parameters, which are also used for mineral oil based fuel and the other group describes the

chemical composition and purity of fatty acid alkyl esters (Mihelbach, 1996).


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Table 1 contains the general biodiesel standards. ASTM 6751-07b is the standard specification for

Biodiesel fuel (B100) blend stock for distillate fuels and summarizes all required tests for B100 Biodiesel

fuel. The monograph consists of the following 20 tests (EMS, 2006):

1.9.1 Calcium and Magnesium

The test method covers the determination of calcium and magnesium in water by complexometric

titration and atomic absorption spectrometric procedures.

1.9.2 Flash point

The flash point is the lowest temperature at which an applied ignition source will cause the vapors of a

sample to ignite. The value of the flash point is used for the classification of flammable and combustible

materials needed for safety and shipping regulations. The flash point is determined by heating a sample

of the fuel in a stirred container and passing a flame over the surface of the liquid. If the temperature is

at or above the flash point, the vapor will ignite and an easily detectable flash can be observed.

Table 1: Biodiesel standards

Property ASTM Method Limits

Flash point (°C) D93 130 minimum

Water & Sediment (vol%) D2709 0.050 maximum

Water in Oil D6304 -

Kinematic Viscosity, 40°C (mm2/s) D445 1.9–6.0

Sulfated ash (% mass) D874 0.020 maximum

Sulfur D5453 –

S 15 Grade (ppm) – 15 max

S 500 Grade – 500 max

Copper Strip Corrosion D130 No.3 maximum

Cetane D613 47 minimum

Cloud Point (°C) D2500 Report

Carbon residue 100% sample (% mass) D4530* 0.050 maximum

Acid number (mg KOH/g) D664 0.50 maximum

Free glycerol (% mass) D6584 0.020 maximum

Total glycerol (% mass) D6584 0.240 maximum

Phosphorus content (% mass) D4951 0.001 maximum

Distillation temperature, atmospheric equivalent temperature, 90% recovered (°C)

D1160 360 maximum

Sodium/potassium (ppm) UOP391 5 max combined

Visual appearance inspection D4176 -

1.9.3 Water and sediment

This method used to measure water and sediment is only sensitive to free water. It uses a centrifuge to

separate small water droplets and particles to be sure they do not exceed 0.05% (500 ppm). This test is

particularly important when working with biodiesel that has been water-washed to remove traces of


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soap and free glycerol. Vacuum drying is usually needed to remove residual water following the washing

process.

1.9.4 Kinematic viscosity

Viscosity is a measure of a fluid’s resistance to flow. The greater the viscosity, the less readily the liquid

flows. The viscosity of petroleum oils is a strong function of temperature with the viscosity decreasing as

the temperature increases. Biodiesel is more viscous than No. 2 petroleum diesel fuel but only by a small

amount. Depending on the feedstock and amount of oxidation, biodiesel viscosity will vary between 4.0

and 6.2 mm2/s, while No. 2 diesel fuels tend to fall in the narrower range of 2.4 to 2.6 mm2/s.

1.9.5 Sulfated ash

When diesel fuel burns, it should be converted entirely to carbon dioxide and water vapor. Inorganic

materials present in the fuel may produce ash that can be abrasive and contribute to wear between the

piston and cylinder. This method measures sulfated ash, which is specified because it is more sensitive

to ash from sodium and potassium. These metals are likely to be the main sources for ash in biodiesel.

1.9.6 Sulfur

The effect of sulfur content on engine wear and deposits appears to vary considerably in importance

and depends largely on operating conditions. Fuel sulfur can also affect emissions, control system,

performance and various limits on sulfur have been imposed for environmental reasons. B100 is

essentially sulfur-free.

1.9.7 Copper corrosion

Many of the compounds in diesel fuel can be corrosive. Copper and copper compounds tend to be

particularly susceptible to chemical attack. In this method, copper strips are placed in the fuel for 3

hours at 50°C. Then the strips are washed in a solvent and compared to a standard description of tarnish

and corrosion.

1.9.8 Cetane number

One of the most important properties of a diesel fuel is its readiness to auto-ignite at the temperatures

and pressures present in the cylinder when the fuel is injected. Fuels with a high cetane number will

have short ignition delays and a small amount of premixed combustion since little time is available to

prepare the fuel for combustion.

1.9.9 Cloud point

When biodiesel starts to crystallize at cooler temperatures, it will also start to clog filters. The critical

temperature depends on the fatty acid composition of the biodiesel. In order to determine the

maximum biodiesel content in a blend for a certain climate it is important to know at which

temperature the biodiesel starts to "cloud". In the test, a sample of Biodiesel is slowly cooled while the

cloudiness is monitored. The cloud point is recorded as the temperature at which the first cloudiness is

observed. The cloud point is the temperature at which a cloud of wax crystals first appears in a fuel

sample that is cooled under conditions described by ASTM D2500.


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1.9.10 Carbon residue

Conradson Carbon Residue Apparatus is used to test petroleum products to determine the amount of

carbon residue left after evaporation and pyrolysis of an oil and to indicate relative coke-forming

propensities.

1.9.11 Acid number

This test method covers procedures for the determination of acidic constituents in petroleum products

and lubricants that are soluble in mixtures of toluene and isopropanol. In B100 (biodiesel), the acid

number is a measure of free fatty acids. The free fatty acids can lead to corrosion and are a symptom of

water in the fuel or fuel oxidation. The sample is dissolved in a mixture of toluene and isopropanol that

contains a small amount of water. The sample is titrated potentiometrically with alcoholic potassium

hydroxide. The meter readings are plotted against the respective volumes of titrating solution and the

end points are taken at well-defined inflections in the resulting curve.

1.9.12 Total and free Glycerol

Gas chromatographic (GC) analysis determines the amount of glycerol (in derivatized form), mono- and

diacylglycerols (both also in derivatized form), triacylglycerols, and methyl esters in a biodiesel sample.

The derivatized glycerol is the first material to elute (free glycerol that has not settled out of solution),

followed sequentially by the methyl esters (biodiesel) and the derivatized monoacylglycerols,

diacylglycerols, and triacylglycerols (intermediate products of the transesterification reaction). This

method gives an indication of the completeness of the reaction and of the quality of phase separation.

1.9.13 Phosphorous content

Phosphorus can damage catalytic converters used in emissions control systems and its level must be

kept low. Catalytic converters are becoming more common on diesel-powered equipment as emissions

standards are tightened, so low phosphorus levels will be of increasing importance. Biodiesel produced

from U.S. sources has been shown to have low phosphorus content (below 1 ppm) and the specification

value of 10 ppm maximum is not problematic. Biodiesel from other sources may or may not contain

higher levels of phosphorus and this specification was added to ensure that all biodiesel, regardless of

the source, has low phosphorus content.

1.9.14 Distillation temperature

The distillation curve is determined by relating the fraction of a fuel sample that is removed by heating a

fuel sample to progressively higher temperatures. However, biodiesel usually only contains 4 to 5 major

compounds that all boil at about the same temperature. In addition, the boiling temperature is so high

at atmospheric pressure that the biodiesel compounds usually decompose (crack) during the distillation

test. Distillation tests following ASTM D86 are not appropriate for biodiesel. The ASTM standard D6751

specifies a distillation test although it recommends ASTM D1160, which is conducted under vacuum.

While this test will allow the biodiesel to be distilled without decomposing, the procedure specified in

the technique for converting the distillation curve back to atmospheric pressure is only valid for

petroleum products and should be used with caution for biodiesel.


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1.9.15 Sodium/Potassium combined

Sodium and potassium may be present in biodiesel as abrasive solids or soluble metallic soaps. Abrasive

solids can contribute to injector, fuel pump, piston, and ring wear, as well as to engine deposits. Soluble

metallic soaps have little effect on wear, but they may contribute to filter plugging and engine deposits.

High levels of sodium and potassium compounds may also be collected in exhaust particulate removal

devices, are not typically removed during passive or active regeneration, and can create increased back

pressure and reduced time to service maintenance.

1.9.16 Oxidative stability

Due to its chemical structure, oxidation rates of FAME can depend on many variables such as

temperature, light, radiation intensity, presence of naturally occurring antioxidants and more. The

oxidation stability of FAME can be increased by adding additional natural and synthetic antioxidants. The

Rancimat™ method expresses the oxidation stability of the tested material in terms of an induction

period for the production of volatile organic acids, which are byproducts of fatty acid ester oxidative

degradation with heat and oxygen.

1.9.17 Cold soak filtration test

When biodiesel gets cold, flaky or greasy precipitates may fall out that can clog filters. These precipitates

are different from gelling in that most of the biodiesel stays liquid while a white flaky or greasy

substance precipitates. The precipitates usually have a high melting point and do not melt back into the

biodiesel at normal temperatures. In this test, biodiesel is chilled to 40°F (~4oC) for 16 h, and

subsequently warmed to room temperature without stirring or heating the sample. It is then passed

through a filter under vacuum. To pass, the full 300 ml sample must flow through the filter in under 360

seconds. The term Cold Soak comes from chilling the biodiesel to 40°F (~4oC) for 16 h.

1.9.18 Visual appearance inspection

Visual inspection indicates successful biodiesel reaction in 2 distinct layers. Top layer: 80–90% product,

lighter color than the bottom layer, it is biodiesel. Bottom Layer: 10–20% of the reaction, darker color, a

mixture of glycerol, catalyst (lye), alcohol & possibly solid particles. A milky middle layer in the test batch

indicates a production of soap (due to water in the oil or too much lye). Unsuccessful reactions result in

no layering or one solid batch of soap.

1.9.19 Water in oil

Moisture is considered a chemical contaminant when suspended or mixed with lubricating oils. It

presents a combination of chemical and physical problems for the lubricant and machinery, respectively.

The effects of water are insidious. Failure due to water contamination may be catastrophic, but it may

not be immediate. Many failures blamed on lubricants are truly caused by excess water. The following

are some of the effects of water on equipment:

Shorter component life due to rust and corrosion;

Water etching/erosion and vaporous cavitation;

Hydrogen embrittlement;

Oxidation of bearing Babbitt;


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Wear caused by loss of oil film or hard water deposits

1.9.20 Workmanship

Free of undissolved water, sediment & suspended matter.

1.10 Glycerol refining, management and utilization Although glycerol is the secondary product from the transesterification reaction, development of

sustainable processes for utilizing this organic raw material is imperative due to its numerous

applications in different industrial products such as moisturizers, soaps, cosmetics, medicines, and other

glycerol products (Wen et al., 2008; da Silva et al., 2009). Its good reactivity on sump oil and its

effectiveness for washing the shearing shed floor is well proven so it can be used as a heavy duty

detergent and degreaser. Table 2 lists the distribution of glycerol consumption in different products and

industries.

1.10.1 Glycerol refining

Since purified glycerol is a high-value and commercial chemical with numerous applications, the crude

glycerol presents a great raw material for glycerol production. Therefore, it is essential to pay more

attention to the utilization of crude glycerol from biodiesel production in order to defray the production

cost of biodiesel and to promote biodiesel on a large scale. Typically, glycerol produced during chemical

transesterification is about 50% glycerol or less in composition and mainly contains water, salts,

unreacted alcohol, and unused catalyst. In addition to that, crude glycerol also contains a variety of

elements such as calcium, magnesium, phosphorous, or sulfur (Thompson and He, 2006).The unused

alkali catalyst is usually neutralized by an addition of hydrochloric or sulphuric acids into the glycerol

phase during the re-neutralization step which results in sodium chloride or potassium sulphate salts

recovery and can be used as a fertilizer (Duncan, 2003). Water and alcohol can be removed later to

produce 80–88% pure glycerol that can be sold as crude glycerol. To achieve 99% glycerol or higher

purity, glycerol is further distilled and sold in different markets (NBB, 2007).

1.10.2 Management and value-added opportunities for crude glycerol

According to today’s scenarios, the global biodiesel market is estimated to reach 37 billion gallons by

2016 with 4 billion gallons of crude glycerol production (Wang et al., 2006). Therefore, it is of great

importance for scientists to find new applications for refined and crude glycerol. Following are the

possibility of utilizing glycerol in different applications.

1.10.2.1 Animal feedstuff

Since 1970s, glycerol has been used as a feed ingredient for animals due to its higher absorption rate

and higher energy source. However, glycerol’s utilization in feeds was limited by the availability of

glycerol (Fisher et al., 1973). Crude glycerol was an excellent source of calories for non-ruminants and

ruminant diets for replacing corn diet. Reported digestible energy values for 85% of the crude glycerol

samples were in the range of 14.9-15.3 MJ/kg with metabolic energy values in the range of 13.9-14.7

MJ/kg (Dasari et al., 2007). Although crude glycerol can be added to animal feed, excess glycerol in the

animal diet may affect normal physiological metabolism.


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In all, the use of crude glycerol as an animal feed component has great potential for replacing corn in

diets, and is gaining increasing attention. However, potential hazardous impurities in crude glycerol from

biodiesel have to be removed as residual levels of potassium may result in wet litter or imbalances in

dietary electrolyte balance while residual methanol is toxic (Cerrate et al., 2006; Douette et al., 2007;

Lammers et al., 2008).


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Table 2: Distribution of glycerol consumption in different products and industries (Bondolini, 2003)

Industry Percent consumption

Cosmetics, soaps, pharmaceuticals 26

Alkyd resins 6

Food and drinks 8

Tobacco 4

Cellulose films 3

Polyglycerol esters 12

Esters 11

Paper 1

Resale 17

Other uses 12

1.10.2.2 Composting

Partially purified glycerol and salts can be used as a compost accelerant and FFA’s act as a weed killer

(Robert et al., 2009). Crude glycerol cannot be composted alone since it is purely a carbon source and

requires adding nitrogen and other nutrients. Also, the compost mixture must also be bulked properly to

allow adequate oxygenation. Due to these limitations, crude glycerol is best suited to co-compost with

other materials such as biosolids, brown and green waste, and bulked manures. Robert et al. (2009)

reported that glycerol-salt mixture and FFA were effective in accelerating composting and in killing

weeds, respectively. However, it requires more vigorous management to assure aerobic conditions of

the compost pile mixed with crude glycerol due to the high BOD/energy content of the crude glycerol. In

addition, there is more risk of a high BOD leachate from the compost pile which should be addressed in

the storm water permit for the co-composting operation (Pyle, 2008).

1.10.2.3 Feed stock for chemicals

Crude glycerol has potential to produce various chemicals through different conversion processes.

Thermochemical conversion: Thermochemical conversion of the crude glycerol produces substances

such as propylene glycol, acetol, and various other products (Pyle, 2008). Crude glycerol can be

converted to acetol using a dehydration reaction and can then be further converted into propylene

glycol through a hydrogenation reaction (Johnson and Taconi, 2007). Studies have also shown that crude

glycerol can be converted to antifreeze at the biodiesel production sites, avoiding extra transportation

(Pagliaro et al., 2007).

Bioconversion: The bioconversion of glycerol is challenging and could possibly be by means of a limited

number of microorganisms (Yazdani and Gonzalez, 2007). However, there is huge potential for

conversion of glycerol to a variety of high value compounds including 1,2 propanediol, 1,3 propandiol

(propylene glycol), lactic acid, methanol, ethanol and hydrogen (Altaras and Cameron, 2000; Saint-

Amans, 2001; Whited et al., 2003; Yun et al., 2003; Ito et al., 2005; Yazdani and Gonzalez, 2007).

Microorganisms such as Escherichia coli, Clostridium butyricum, Enterococcus faecalis, Enterobacter

aerogens and their recombinant strains have shown potential to convert glycerol into 1,3 propandiol


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(propylene glycol) (Whited et al., 2003), 1,2 propanediol (Altaras and Cameron, 2000), lactic acid (Yun et

al., 2003), methanol (Saint-Amans, 2001), ethanol and hydrogen by aerobic and/or anaerobic

fermentation processes (Ito et al., 2005). However, a detailed bioprocess study would be required to

develop feasible fermentation techniques for conversion of glycerol into a multitude of high value

compounds.

Catalytic conversion: The crude glycerol can be catalytically converted into substances such as methyl

acetate, acrolein, hydrogen or syngas, monoglycerides and various other products. Methyl acetate (2,2-

dimethyl-1,3-dioxolan-4-yl) is used as a fuel additive specifically in biodiesel (García et al., 2008).

Hydrogen or syngas can be produced from crude glycerol via gasification (Yoon et al., 2010). Acrolein is

an important starting chemical for producing detergents, acrylic acid ester and super absorber polymers

(Sereshki et al., 2008).

1.10.3 Disposal

Combustion and landfilling are the proposed disposal methods for glycerol generated from biodiesel

production. However, there is very insignificant research on the land application of crude glycerol.

Studies reported that land application is the least desirable and hazardous option due to its high salt

content, methanol and BOD concentration and has no fertilizer value. Guidelines for the disposal of

grease trap wastes suggests incorporation and loading rates of about 4 tons per acre as appropriate to

avoid choking of soils. Essentially, for land application to be benign, loading rates must be low, and

runoff control is absolutely essential (Pyle, 2008). Disposal into the public sewer is also not advisable

due to very high BOD (10,000- 100,000 mg/L) levels and can easily cause organic shock in the biological

process of a sewage treatment plant. Combustion of crude glycerol is a method that has been used for

disposal. However, this method is not economical for large producers of biodiesel (Johnson and Taconi,

2007).


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2 C-ASD approach for Biodiesel Production The Centre for Aquaculture and Seafood Development (CASD) is proposing to establish a full range of

biodiesel research capabilities within the Marine Institute and Memorial University, and to develop a

proof of concept for fish oil derived biodiesel production system suitable for the unique situation in

Newfoundland and Labrador. The biodiesel processing system recently installed at the Marine

Institute’s by-products facility was purchased from Biodiesel Logic Ltd. with funding support from the

Department of Fisheries and Aquaculture. It is the only system of this kind in Atlantic Canada providing

CASD with the capability of processing oil feed stocks in batches ranging from 60 – 209 L (16 – 55

gallons).

2.1 Project scope This project was built on past projects to advance current biodiesel conversion technologies for specific

applications to marine waste oil feedstock in rural communities. Existing conversion technologies have

been developed to handle primarily homogenous oils from vegetable sources. These technologies have

had limited success when applied to marine waste oils. Mitigating the technical challenges specifically

associated with utilizing marine waste oils as biodiesel feedstock will be the focus of the proposed

project. The long term objective of the proposed research is to develop an economically viable, small to

medium scale fish oil derived biofuel/biodiesel production system for salmon aquaculture

communities located in rural areas of Newfoundland and Labrador. This will enable aquaculture growers

and processors to utilize their waste streams to produce biodiesel and operate feed barges, marine

vessels and generators located at their remote locations. The short-term objectives are: (a) compiling

information on the sources and amounts of fish processing wastes in Newfoundland and Labrador, (b)

characterization of the wastes (c) investigation of mechanical homogenization, heat extraction and

centrifugation techniques that will increase yield of oil during the lipid extraction process (d) assessment

of the waste fish oil quality for use as biodiesel (e) technical evaluation of the degree of processing

required for the waste fish oil to be used effectively in pilot scale biodiesel production (f) evaluation of

methanol recovery during the transesterification process (g) economic evaluation of the biodiesel

produced from waste fish oils as compared with biodiesel produced from plant oils, and other waste oils

of biological origin.

2.2 Deliverables for year 1 • To determine the quantity of marine oil potentially available as biodiesel feedstock in the

province.

• To assess the quality of waste fish oil for use as biodiesel feedstock extracted from various wild

capture fisheries and aquaculture sources.

o Determine the chemical composition, stability, reactivity, and partitioning of the fish waste

o Determine recoverable oil and the chemical composition and stability of the waste fish oil


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o Technically evaluate the degree of processing required for the waste fish oil to be used effectively in diesel engines and/or as heating oil.

• To evaluate the economics of biodiesel produced from waste fish oils as compared with biodiesel produced from plant oils, and other uses of the waste oil.

• To develop simple and effective methods to produce biodiesel from waste fish oils.

2.3 Previous challenges In previous projects, salmon, cod liver, and seal oil have been used to produce marine biodiesel on

small and pilot-scale. Throughout these projects, several problems have been encountered during the

chemical transesterification reaction. However, there were limited necessary personnel, material and

analytical resources available to develop a complete solution to these problems.

The following challenges were identified during previous transesterification experiments:

Low biodiesel yields

Incomplete transesterification reaction

Extremely slow phase separation (biodiesel and glycerol) after chemical transesterification

Very small glycerol droplets

Incomplete methanol recovery

Lack of information on oil quality and pretreatment requirements

Limited availability for oil and biodiesel characterization methods

To address the above mentioned challenges, small scale experiments for oil pre-treatment and biodiesel

production were carried out during year one before conducting the pilot scale biodiesel production.

2.4 Small scale experiments In order to minimize usage of feedstock oil and waste generation during biodiesel production,

optimization of pretreatment and chemical transesterification parameters were carried out on a small

laboratory scale and have been further used for pilot scale biodiesel production. A number of small

scale experiments were carried out in year one in order to investigate the challenges mentioned above

that were encountered during the transesterification of marine oils in previous projects according to the

traditional recipe for waste vegetable oils. The influence of several pre-treatment and reaction

parameters on the outcome of the chemical transesterification was investigated, in particular:

Free fatty acid reduction prior to transesterification

Degumming of marine oils prior to transesterification

Modification of the Methanol : Oil molar ratio during transesterification

Modification of the amount of catalyst used during transesterification

These parameters have been investigated on a small scale using seal and salmon oil; the corresponding

experiments and detailed results are attached in Appendix I. Salmon oil transesterification could be

improved significantly by modifying the above mentioned parameters. Based on the findings, a pilot-


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scale process could be developed which is presented in Section 2.5. The seal oil reaction could be

somewhat improved but more small scale research is necessary in order to propose a suitable pilot scale

process.

Large scale oil extraction could not be performed in year one of the project due to incomplete set-up of

pilot scale oil extraction facilities. In order to proceed with small scale experiments, seal and salmon oil

was purchased. The oil was selected according to the following criteria:

Oil extracted from Atlantic salmon or Harp seal

Atlantic salmon oil should originate from Newfoundland aquaculture

Oil should be recovered through heat extraction with an equipment set-up similar to the oil

extraction line to be installed at the bioprocessing facility

880 L industrial grade salmon oil (extracted in April 2012 and stabilized with antioxidants) and 1200 L

industrial grade seal oil were received at the bioprocessing facility on 14 May 2012. In addition, 500 kg

seal blubber were sourced and received. Complete characterization of marine oil was performed in

order to identify pretreatment steps required to facilitate the biodiesel production.

2.4.1 Oil refining

Procured oil characterization has been performed and found that the amount of phospholipids in

salmon and seal oil was higher (1.5%) than the recommended maximum concentration (0.5-1.0%).

Degumming with water effectively removed phospholipids from salmon and seal oil which has shown

significant improvement in salmon oil transesterification and certain positive effects on seal oil

transesterification. Phase separation, size of glycerol beads and reaction speed were significantly

improved during transesterification after implementation of the hydration step, in the case of salmon

oil. Next, in order to determine the percentage of FFA in the oils or fats, titration was performed and

found that the percentage of FFA was 2.25% (within the range of 0.5-2.5 wt%). Therefore, no

pretreatment is necessary to reduce the content of FFA (For more details refer section 4). It has been

reported that the high amounts of unsaturated fatty acids lead to oxidative spoilage of oil and biodiesel

and require the addition of antioxidants. Marine oil analysis has shown that both salmon and seal oil

contain high amounts of unsaturated fatty acids and therefore, an anti-oxidant designed to stabilize fish

oil and biodiesel has been sourced.

2.4.2 Chemical transesterification

The chemical transesterification process is highly sensitive to the reaction conditions, including:

pretreatment, alcohol to oil molar ratio, mixing intensity type, amount of catalyst and temperature. The

alcohol to oil molar ratio was increased from 6:1 to 12:1 and has shown significant improvement in

transesterification reaction time and glycerol and biodiesel phase separation. Further, optimization of

the alcohol to molar ratio for salmon and seal oil is required. Oil analysis suggested that the water

content in seal and salmon oil was below (500 ppm) compared to the allowable critical limit (1500 ppm).

However, drying of the oil prior to transesterification was implemented and shown to improve the

transesterification reaction time and biodiesel and glycerol phase separation.


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It was identified that the NaOH used was not stored completely sealed and thus did not maintain

anhydrous form. To avoid the soap formation during transesterification step due to moist NaOH, a new

lot of NaOH was purchased for future experiments. It was found that the variations in mixing speed and

stirrer shape did not have any significant influence on the reaction in terms of phase separation and size

of glycerol beads.

2.5 Proposed pilot scale fish oil and biodiesel production The second step in the development of CASD’s biodiesel production process is the scale up of the

laboratory process to a pilot scale. The ability to perform pilot-scale experiments in year one of the

project has been limited due to ongoing small-scale research, limitations in fresh oil supply, and a

shipping delay of the pilot scale oil extraction equipment.

Based on literature research and the findings from the small scale experiments, a process for biodiesel

production could be designed which is able to process salmon oil. In this chapter, the general approach

and the main equipment is described. Figure 2 shows the schematic diagram of the proposed biodiesel

production process including heat extracted oil recovery and pretreatment as developed specifically for

locally available marine oil feedstock. The process for salmon oil derived biodiesel production described

below has been established based on one-batch pilot-scale experiment which showed promising results.

The research plan for pilot-scale biodiesel production includes:

(a) Scale-up of the pre-treatment and transesterification process developed in the laboratory

(b) Assessment of solvent recovery and reuse

(c) Recovery of partially crude glycerol

(d) Technical evaluation of the performance of fish oil derived biodiesel for use in diesel

engines/boilers/generators etc. by using a dynometer located at MUN Engineering.

The raw materials required for oil extraction will be provided by the CASD’s industry partners (i.e. Cooke

Aquaculture, Seaward Farms Ltd., and Allen’s Fisheries Ltd.). The alcohol used in the transesterification

process in this project is methanol due to its small molecular size and fast reaction rates. However, other

primary and secondary monohydric aliphatic alcohols having 1-8 carbon atoms could be used. Sodium

Hydroxide is used as a catalyst to carry out the transesterification process.

2.5.1 Fish oil extraction

2.5.1.1 Heat extraction

Heat extraction has been selected as the method of choice for the extraction of oil from salmonid waste

streams as it is well established and equipment is readily available for small, medium and large scale

productions. Two pieces of equipment essential for heat extraction (Hobart™ meat grinder and

Westphalia centrifuge) are already installed and functional at the CASD bioprocessing facility. In

addition, a Contherm™ scraped surface heat exchanger and a decanter centrifuge have been purchased

from Alfa Laval in year one of the project.


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Figure 2: Schematic flow chart for the proposed Biodiesel production process


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Installation of this equipment will take place in April 2013 and will allow CASD to set-up an oil extraction

line with a processing capacity of 1000 kg/hour of fish waste, resulting in about 200-250 L oil per hour

for use in biodiesel pilot scale studies. Following is a description of the main pieces of equipment and

their function for heat extraction in sequence of use.

Hobart™ meat grinder: The first step in oil extraction from fish processing waste is the homogenization

of fatty tissues and bones and skin which prepares the material for the oil extraction by heat treatment

or enzymatic hydrolysis. Homogenization can be achieved at the bioprocessing facility using the

Hobart™ meat grinder (Figure 3), which reduces the particle size to 3 mm and which can process up to

1000 kg of raw material per hour. The grinder is installed and functional at the bioprocessing facility.

Contherm scraped-surface heat exchanger: The Contherm™ is a cylindrical scraped-surface heat

exchanger (Figure 4) with a rotating blade inside which distributes the feed stream evenly across the

heated surface while it passes through the cylinder. The material is heated and the fat cells burst and

release the oil into the solution. The Contherm allows continuous processing of large volumes of ground

fish meat, with a residence time of about 2 min. The equipment has been purchased and arrived at the

bioprocessing facility in March 2013. It will be installed and commissioned in April 2013.

Three-phase decanter: A decanter centrifuge (Figure 5 and Figure 6) is a cylindrical shaped centrifuge

with a rotating spiral inside. As the cylinder spins around its longitudinal axis, the solid particles are

pressed against the inside wall of the cylinder. The spiral scrapes off those solids and transports them to

the solid discharge point while the liquid phases are discharged through ports closer to the core of the

decanter. The equipment has been purchased and will be installed at the bioprocessing facility in April

2013.

Westphalia centrifuge: Oil obtained after the three phase decanter centrifuge is relatively moist; hence

it becomes essential to dry the oil before proceeding with biodiesel production. Drying of the oil can be

achieved by centrifugation at high speeds in a Westphalia centrifuge (Figure 7), which separates oil and

water effectively. The equipment is already installed at the bioprocessing facility and ready to use.

2.5.1.2 Ensilaging

Ensilaging of fish waste has been identified as a possible second option for oil extraction as waste

streams could be processed quickly, in large volumes and with minimal process control, while the oil

could be converted to biodiesel as the reactor capacity allows. In year one, no activities have been

performed in terms of silage production. The ensilaging process would involve placing the fish waste

into a large silage tank equipped with heavy duty pumps for fish waste recirculation. Conventionally, 5 %

(v/v) formic acid (or another suitable acid) is added to the fish waste in order to lower the pH for

stabilization against spoilage. Then, the mixture is incubated for 2-3 weeks with occasional mixing until

the endogenous enzymes present in the gut material of the fish can degrade the tissue and bone

material of the fish waste. The reaction speed is dependent on the surrounding temperature and

generally not controlled. The downstream processing, i.e. the isolation of the oil after release from fat

cells, can be done using the decanter centrifuge and Westphalia centrifuge as described in the previous

Section 2.5.1.1.


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Figure 3: Hobart grinder.

Figure 4: Contherm scraped-surface heat exchanger


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Figure 5: Decanter centrifuge

Figure 6: Working principle of a decanter centrifuge


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Figure 7: Westphalia centrifuge for oil polishing


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2.5.2 Pre-treatment of oil

It is very essential to characterize each batch of oil extracted from marine wastes in order to identify

optimal pre-treatment conditions and achieve a successful chemical transesterification reaction.

Generally crude marine oil contains triglycerides, free fatty acids, water, phospholipids and other

contaminants in various proportions. There are numerous pre-treatment techniques, however, only a

few are important for preparing oil for chemical transesterification. The stabilization of oil, degumming,

FFA reduction and glycerol stripping has been identified as steps very likely to be included in the

production process.

Further research in year two will establish the pre-treatment for salmon and seal oil depending on the

outcomes of pilot scale oil extraction.

2.5.2.1 Oil stabilization

“Oxidative rancidity” can occur in fish oil due to the high amounts of unsaturated fatty acids and natural

oxidation rate during storage and extraction process. It is possible to retard the development of

oxidative deterioration, by incorporating a suitable antioxidant at early stages of oil extraction which is

capable of stabilizing the substrate. Anti-oxidant is added to the extracted oil immediately after every oil

extraction experiment performed.

2.5.2.2 Degumming

Phospholipids can be hydrophobic or hydrophilic and have an emulsifying effect in the chemical

transesterification reaction. Degumming is the process of removing phospholipids from oil. There are

different degumming methods, such as membrane filtration, hydration, acid micelles degumming and

supercritical extraction. In the hydration process, hot water is added to the oil with stirring and the

water soluble portion of the phospholipids will dissolve in the water. The water can be removed from

the oil by natural settling and by evaporating the residual water. The hydration method is a simple,

easy and high yields refining method. It is recommended for small to medium scale producers.

However, non-hydratable phospholipids cannot be removed using this method. Degumming by

hydration is performed in a separate pre-treatment tank in which the oil can be heated to the optimal

reaction temperature of 55-60°C. A pump recirculates the oil to ensure mixing of water and oil. After 1

h reaction time, the recirculation pump is turned off and the water is allowed to settle out of the

mixture at the bottom of the conical tank for 12 h followed by draining.

2.5.2.3 Reduction of FFA content

Free Fatty acid contents above 2.5 wt% in oil cause an increased soap formation during

transesterification and biodiesel yields are greatly reduced. In order to determine the percentage of FFA

in the oils, titration is performed. If the percentage of FFA is over 2.5 wt.%, an additional acid

esterification step may be necessary to reduce the content of FFA. Esterification by acid-catalysis makes

the best use of the free fatty acids in the oil and transforms it into biodiesel. In year one, the results of

oil analysis and small scale experiments have indicated that an acid esterification may not be necessary

for salmon and seal oil however, these findings need to be confirmed in year two.


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2.5.2.4 Glycerol stripping

An additional pre-treatment step, glycerol stripping is applied further to degumming. The fish oil is

mixed with partially crude glycerol (which is a mixture of glycerol, unreacted methanol and sodium

hydroxide) obtained from the previous batch of chemical transesterification. The catalyst (sodium

hydroxide) reacts with the free fatty acids present in the oil and reduces the acid value before the main

chemical transesterification reaction. After 1 h reaction time at 55 to 60°C, the recirculation pump is

turned off and the mixture is allowed to settle at the bottom of the conical tank for 2 h followed by

glycerol draining. This treatment has been shown to be good practice by many biodiesel producers and

is an integral part of the biodiesel production line purchased from Bio-Diesel Logic. There is no plan to

perform extensive research on this particular step.

2.5.3 Chemical transesterification (Biodiesel production)

Chemical transesterification is carried out using an equipped two-tank system in the biodiesel

production line. In the reaction tank, the oil is pre-heated to the optimal reaction temperature of 55 to

60°C, while the methoxide is prepared in a separate methoxide preparation tank according to the

developed recipe. The methoxide is injected into the reaction tank and mixture is allowed to react for 1

h with continuous mixing. After 1 h reaction time, the mixing is stopped and the mixture is allowed to

settle for 2-12 hours in the reaction vessel in order to allow the initial separation of biodiesel (top phase)

and partially crude glycerol (bottom phase). The glycerol is drained from the tank and stored for the pre-

treatment of the next batch of pretreated fish oil.

2.5.4 Methanol recovery

Following the partially crude glycerol separation, the raw biodiesel is heated to 80°C for 1 h while

stirring continuously on order to evaporate the methanol dissolved in the fuel. The methanol vapors are

condensed and recovered for use in methoxide preparation for the next batch.

2.5.5 Biodiesel purification

While biodiesel purification on a small scale is achieved using water washing, pilot-scale processes use

dry wash systems. In this project, a built-in dry wash resin Amberlite™ is used to remove free glycerin,

soap and other impurities from the biodiesel.

In addition, several filtration steps performed before and after pre-treatment as well as after biodiesel

purification using dry washing ensure the production of highly pure biodiesel suitable for cold climate

use.

2.6 Bio-Diesel Logic™ production line The bio-diesel processing system purchased by CASD can process 220 L of oil per batch (Figure 8 (a) and

(b)). The system, manufactured by Bio-Diesel Logic, incorporates strategies to improve the efficiency of

the system through pre-treating the incoming raw oil and methanol recovery.

This system has been designed for processing waste vegetable oils and includes guidelines on optimal

transesterification conditions and oil pre-treatment. The nature of marine oils and their specific needs


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for pre-treatment and transesterification required several modifications to the line and some recipe

changes which are described in Chapter 3. The production line (as purchased) includes a tank for pre-

(a) Front view

(b) Back view

Figure 8: Demonstration biodiesel production line


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treatment for the oil (Glycerol stripping), a reaction tank, a methoxide mixing tank, a fuel storage tank, a

dry wash column and a ColdClear™ fuel filtration system.

The specifications of the whole system are attached as Appendix IV. A detailed description of the piping

and pump set-up can be found in Chapter 3.

2.6.1 Pre-processing tank

This stainless steel tank has 316 L of capacity (312L tank + 16 L cone bottom) and is equipped with an

integral sight gauge for visual indication of tank liquid level and stainless steel alloy heater assembly. The

reactor vessel is insulated with 1/4" high density ceramic fiber insulation and wrapped with a hammered

finish aluminum cover.

2.6.2 Reaction tank

The reaction tank has the same specifications as the pre-processing tank.

2.6.3 Methoxide tank

This 50 L HDPE methyl/oxide mixing tank is equipped with high speed methanol and sodium hydroxide

propeller mixer with stainless steel shaft. Based on feedstock oil type and characteristics, the desired

proportion of methanol and sodium hydroxide mixture is prepared in this tank prior for injection into

the reaction tank.

2.6.4 Fuel storage tank

This 400 L carbon steel fuel storage tank is used for storage of biodiesel produced after the chemical

transesterification process.

2.6.5 Dry wash system

The dry wash system is equipped with 88 lb Amberlite™ BD10 dry wash media (ion exchange resin) to

process a minimum of 10,000 gallon of biodiesel before introducing it to the ColdClear™ filtration

system. This system helps to bring the free glycerol level down and removes residual soaps.

2.6.6 ColdClear filtration

ColdClearTM is a three stage system with all housings mounted in series on a single skid. The first stage

serves as a pre-filter and captures solid particulates down to three microns using high efficiency

Excellement® catridges. Stages 2 and 3 utilize cartridges that combine adsorption technologies with the

proven effectiveness of Schroeder’s high efficiency Excellement® synthetic media. This multi-stage

filtration/adsorption system ensures any potential factors that would initiate crystallization or plugging

on the filter are dramatically reduced by sequentially removing certain impurities that create a higher

than normal likelihood of surface crystallization on the filter. This technology ensures that produced

biodiesel can meet the ASTM specification for cold soak filtration.

2.7 Generator installation at Sugarloaf aquaculture site, Bay d’Espoir A diesel generator was installed at the Sugarloaf aquaculture site in Bay d’Espoir (Figure 9) for the

purpose of performance testing of biodiesel produced at CASD’s bioprocessing facilities. A brand new 10

hp Kubota generator was first shipped to the bioprocessing facility in May 2010 for general maintenance


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Figure 9: Generator installation at Sugarloaf aquaculture Site, Bay d'Espoir


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and test runs before it was brought to the South coast of Newfoundland. The generator was installed on

the barge of the cabin that houses 4 workers during the summer months. Once pilot scale production

begins, biodiesel will be used in blends (B10 or B20) to provide general utilities including heat, hot water

and electricity to the cabin. After review of the electrical equipment in the cabin, it was decided to

purchase additional equipment (baseboard heaters, hot water tank) to provide an optimal load of 80%

on the generator. The generator was run with regular diesel during the summer to establish baseline

data.

For recording data on-site, forms have been developed that will be filled out by site personnel (Appendix

V). Several pieces of equipment have been procured and prepared for installation including a power

meter to record the kWh used and temperature data loggers for recording the exhaust, oil and ambient

temperatures.


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3 Pilot-scale Process Design

Due to specific characteristics of marine oils, additional pre-processing steps (degumming or acid

esterification) become necessary. Therefore, an additional tank will be added to the existing biodiesel

processing line, which will allow the inclusion of an additional pre-treatment step.

Figure 10 shows a schematic of the processing line as purchased with the proposed

modifications/additions to the processing line shown in red. Table 3Error! Reference source not found.

provides a detailed description of the processing line components. The purchase of a new tank and its

installation to the biodiesel production line is part of year two of the project.

At this point, the modifications proposed only reflect the positive results obtained for pretreatment and

chemical transesterification of salmon oil. Further modifications may be necessary in terms of the seal

oil pretreatment and biodiesel production which needs to be further established in year two.

For the design of the pilot-scale processing line, the following factors were considered:

Selection of equipment that can be easily transferred to industrial scale

Safety of equipment

Minimal manual handling of chemicals, product, by products and waste

Short reaction times

Easy operation of the process line


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Figure 10: Process design of demonstration biodiesel production line


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Table 3: Process design of demonstration biodiesel production line

System component Description and function

Oil barrel Oil storage barrel.

Pump I Hand pump. Used for pumping the oil from the barrel into Tank I.

Filter I Size 3 carbon steel WVO pre-filter canister with a 400 µm polyprolylene felt bag filter. Prevents course and fine impurities of oil from entering Tank I.

Valve I Valve controlling the inlet into Tank I.

Tank I 84 gal (320 L) insulated steel tank with interior heating element and cone bottom. Used for water degumming of oil as pre-treatment for chemical transesterification.

Heater, Tank I 2250 W, stainless steel alloy heater

Valve II Valve controlling the drain from Tank I. Used for draining off water after the degumming step.

Valve III Valve controlling the outlet of Tank I.

Pump II 1/2 hp, 1 Ph mixing pump. Used to re-circulate the oil in Tank I and to transfer the oil from Tank I to Tank II.

Switch I Switch I is used to control whether the oil is re-circulated in Tank I or whether it is pumped into Tank II.

Valve IV Valve controlling the inlet into Tank II.

Tank II 84 gal (320 L) insulated steel tank with interior heating element and cone bottom. Used for treatment of the oil with the glycerin produced during the previous batch transesterification.

Heater, Tank II 2250 W, stainless steel alloy heater

Valve V Valve controlling the outlet of Tank II.

Pump III 1/2 hp, 1 Ph mixing pump. Used to re-circulate the oil in Tank II and to transfer the oil from Tank II to Tank III.

Switch II Switch II is used to control the drain of Tank II for extracting oil samples for analysis before the transesterification reaction and for draining off the used glycerin after pre-treatment.

Switch III Used to switch between oil re-circulation in Tank II and oil transfer into Tank III.

Filter II Size 3 carbon steel WVO pre-filter canister with a 200 µm polypropylene felt bag filter. Prevents particles to enter Tank IV.

Valve VI Valve controlling the oil inlet to Tank III

Tank III 84 gal (320 L) insulated steel tank with interior heating element and cone bottom. Used for the transesterification reaction.

Heater, Tank III 2250 W, stainless steel alloy heater

Valve VII Valve controlling the outlet to Tank III.

Tank IV 15 gal (57 L) high density poly ethylene tank for Methoxide preparation.

Stirrer 1/3 hp, 1 Ph high speed prop mixer for Tank IV with stainless steel shaft.

Pump IV Pneumatic methanol/methoxide injection pump. Used to pump (inject) Methoxide from Tank IV into the oil-recirculation cycle in Tank III.

Valve VIII Valve controlling the methoxide inlet to Tank III

Condenser Methanol condenser. Cools the evaporated Methanol vapors.

Pump VI 1/3 hp pneumatic methanol transfer pump. Used to pump the condensed Methanol back into Tank IV.


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Pump V

1/2 hp, 1 Ph fuel transfer pump. Depending on the setting of switches IV and V, this pump is used to (a) transfer the raw biodiesel from Tank IV into Tank V (fuel storage), (b) transfer the glycerin to Tank II or (c) to re-circulate the transesterification reaction mixture in Tank IV.

Switch IV Switch IV controls the transfer of the glycerin to Tank II and the transfer of the fuel to the storage tank (Tank V).

Switch V Switch V controls the recirculation of the fuel/reaction mixture in Tank III.

Tank V Fuel storage tank

Pump VII 1/2 hp, 1 Ph fuel transfer pump. Used to transfer the fuel from Tank IV to Tank V.

Filter III Size 3 carbon steel WVO pre-filter canister with a 5 µm polyprolylene felt bag filter. Prevents fine particles from entering the Dry Wash column.

Dry wash column

Filled with 88 lbs Amberlite BD10 dry wash media. Used to remove soap, catalyst residues and other impurities from the fuel.

Filter IV ColdClear™ filtration system. To meet the ASTM specification for cold soak filtration biodiesel test

Note: Components in red are proposed modifications to the purchased biodiesel production line which will be installed in year two of the project.


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4 Feedstock Oil and Biodiesel Characterization A proximate analysis of the fish waste streams was performed in order to identify the theoretical

recoverable amounts of oil. Characterization of fish oil allowed making a statement about its suitability

for biodiesel production. A literature review has been performed in order to identify necessary tests that

have to be performed on feedstock oil prior to biodiesel conversion and quality of biodiesel for

standardization of fuel quality. The aim of the literature review was to define a number of tests that can

be performed with a minimally equipped on-site laboratory and provide a good indication on the quality

of the biodiesel batch to the producer before its use in a diesel engine.

Suggested test methods and results obtained in year one on fish oil and biodiesel characterization are

summarized in the following sections.

4.1 Proximate analysis of salmon waste streams A proximate composition of the salmon waste streams is shown in Table 4. It has been found that

salmon guts and frames have similar amount of lipid contents.

4.2 Feedstock oil characterization It is essential to characterize each batch of feedstock oil prior to pre-treatment and chemical

transesterification due to its varying characteristics. Feed stock oil was analyzed after refining and

before chemical transesterification. While some procedures require extensive preparation and high

investments for equipment and a laboratory facility, some tests are quick and easy and give a good

indication on the quality of the feedstock oil.

The procedures identified and incorporated for onsite fast testing have been selected with consideration

of reliability, easiness and minimum use of equipment. The procedures used for oil analysis are

described in Appendix II. Analytical methods applied for characterization of marine oils are shown in

Table 5. Analytical methods described in Section 1.5 have been applied to characterize the purchased

seal and salmon oil for small scale experiments in year one. Table 8 shows test results obtained for some

small scale experiments in year one. Further findings are described in Appendix I.

4.2.1 Appearance and smell

Generally salmon oil contains more pigment compared to seal oil and results in an orange color. Oil

should not contain any course or fine impurities and rancid smell. The salmon and seal oil procured from

Newfoundland’s producers have fulfilled all quality requirements in terms of appearance and smell. The

evaluation of appearance and smell will be of greater importance in terms of pilot scale oil extraction (in

year two).

4.2.2 Free Fatty acids

The free fatty acid contents in both seal and salmon oil were below 2.5% (Table 88). These results were

unexpected as several scientific papers on marine oil biodiesel production identified high FFA contents

in oil as the primary challenge.


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Table 4: Proximate analysis of fish waste streams

Parameters Guts Heads Frames/trimmings

Moisture 64.52 62.88 57.07

Lipids 20.32 23.59 23.73

Protein 9.59 9.59 14.52

Ash 1.14 3.90 4.65 *All values are based on wet weight

Table 5: Analytical methods applied for characterization of marine oils

Analytical methods for oil

ID Test parameter This test can be performed at:

Relevance for Biodiesel production

Performed for every batch of feedstock oil?

1 Appearance and smell MI High Yes

2 Free Fatty Acids MI High Yes

3 p-Anisidine value MI High Yes

4 Peroxide value MI High Yes

5 Water content MI High Yes

6 Fatty acid profile MUN (OSC) High No

Table 6: Fatty acid profile of procured seal and salmon oil

Seal oil Salmon oil

% Lipid Composition Sample 1 Sample 2 Average Sample 1 Sample 2 Average

Hydrocarbons 0.00 0.22 0.11 0.00 0.00 0.00

Steryl Esters/Wax Esters 0.00 0.23 0.12 0.00 0.00 0.00

Ethyl Esters 0.00 0.00 0.00 0.00 0.00 0.00

Methyl Esters 0.00 0.00 0.00 0.00 0.00 0.00

Ethyl Ketones 0.00 0.00 0.00 0.00 0.00 0.00

Methyl Ketones 0.00 0.00 0.00 0.00 0.00 0.00

Glycerol Ethers 0.00 0.00 0.00 0.00 0.00 0.00

Triacylglycerols 89.37 95.08 92.22 92.38 91.29 91.83

Free Fatty Acids 1.37 1.22 1.29 2.29 2.38 2.33

Alcohols 0.00 0.00 0.00 0.00 0.00 0.00

Sterols 3.41 1.25 2.33 3.68 3.60 3.64

Diacylglycerols 0.00 0.00 0.00 0.00 0.00 0.00

Acetone Mobile Polar Lipids 3.78 1.06 2.42 0.64 0.89 0.77

Phospholipids 2.08 0.94 1.51 1.00 1.85 1.43


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4.2.3 Peroxide and p-Anisidine value

The peroxide value is a measure of primary degradation products, while the p-Anisidine value is a

measure of secondary degradation products. The formation of primary and secondary degradation

products will be prevented by using a stabilizing agent (antioxidant), which is added to the feedstock oil

immediately after oil extraction. However, these two tests will be performed on oil before chemical

transesterification, especially if the oil has been stored for some time before it is used in the biodiesel

production process.

4.2.4 Water content

The water content can be measured using an alternative method (not according to ASTM) offered as a

test kit by Sandy Brae Inc (Appendix III). The test kit allows accurate measurement of water levels in

feedstock oil prior to transesterification. The oil should be as dry as possible to avoid soap formation and

improve phase separation and biodiesel yields. The purchased seal and salmon oil had very low water

contents (<500 ppm), nevertheless, an oil drying step prior to transesterification is always included.

4.2.5 Fatty acid profile

The fatty acid profile of the purchased seal and salmon oil was analyzed and it was found that the

amount of phospholipids in salmon and seal oil was higher (1.5%) than the recommended max

concentration (0.5-1.0%), indicating that a degumming step (i.e. removal of phospholipids) is necessary.

The free fatty acid (FFA) content was low for both oil feedstocks (Seal: 1.29% and Salmon: 2.33%),

indicating that an additional pre-treatment for removal of FFA is not necessary. Another significant

difference between the two oil feedstocks was observed in the concentration of Acetone Mobile Polar

Lipids (AMPL’s). AMPL’s was significantly higher in seal oil (2.42%) compared to salmon oil (0.77%).

Table 6Error! Reference source not found. shows the results of the fatty acid analysis. Fatty acid

profiling will not be performed on every batch of feedstock oil produced. However, seasonal changes in

fatty acid profile will be observed to make adjustments to the reaction parameters if needed.

4.3 Biodiesel characterization Unlike conventional diesel fuel, biodiesel is a 100% renewable fuel and it has to fulfill the same

specifications like conventional diesel fuel. The latest specification for B100 biodiesel fuel is ASTM

D6751-07b. ASTM is a standards group comprised of engine and fuel injection equipment

manufacturers, fuel producers, and fuel users whose standards are recognized in the U.S. by

governmental entities, including state agencies responsible for ensuring fuel quality. So-called “biofuels”

or “biodiesel fuels” that do not meet the ASTM standard outlined are not legally biodiesel fuels and

should not be used in diesel engines. Produced biodiesel must be tested in an accredited laboratory to

verify it meets specifications. This can be expensive.

Few specifications including flash point and cloud point are very critical while others including sulfur,

phosphorous residues and carbon residue are only rarely a problem with biodiesel. It was furthermore

identified that some tests can be replicated with minimum equipment in a “homebrew” laboratory,

while others can only be done using professional equipment and specialized technicians for their

operation.


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In this chapter, a testing monograph is suggested for pilot-scale biodiesel production that

(a) Minimizes the need for analysis through an accredited laboratory and

(b) Makes a qualitative statement on the biodiesel quality and its safety for use in a diesel

generator.

Table 7 presents the proposed testing monograph for biodiesel produced on a pilot scale. Table 8 shows

test results obtained for some small scale experiments in year one.


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Table 7: Suggested testing monograph for biodiesel produced on pilot scale

ID Test parameter ASTM # This test can be performed at:

Relevance for Biodiesel testing

Tests performed for every batch of biodiesel produced

1 Flash point ASTM D93 MI High

2 Water SandyBrae test (alternative test method, not ASTM approved)

MI High

3 Kinematic viscosity, 40°C

ASTM D445 MI High

4 Cloud point Modified ASTM method/homebrew method MI High

5 Acid number ASTM D664 MI High

6 Total and free glycerol

ASTM D6584 MI High

7 Cold Soak filtration test

Modified ASTM method MI High

8 Workmanship MI High

9 pH Standard laboratory test MI High

10 Reaction completion Homebrew method MI High

11 Soap value Homebrew method MI High

12 Density Standard laboratory test MI High

13 Emulsification Homebrew method MI High

Tests performed seasonally

14 Oxidative stability ASTM EN14112 accredited laboratory

High

15 Sulfated ash Modified ASTM method MI Medium

16 Sulfur ASTM D5453 MUN/ accredited laboratory

High

17 Copper corrosion Modified ASTM method MI Medium

18 Carbon residue ASTM D4530 MI Low

Tests performed in process design phase only, afterwards as annual control

19 Calcium and Magnesium

ASTM EN 14538 MUN Low

20 Cetane number ASTM D613 accredited laboratory

Low

21 Phosphorous ASTM D4951 MUN/accredited laboratory

Low

22 Distillation temperature

ASTM D1160 accredited laboratory

Low

23 Sodium/Potassium combined

ASTM EN14538 MUN/accredited laboratory

Low

*Laboratories at MI and MUN are not accredited and procedures and equipment differ slightly from the official ASTM method. However, some tests can be performed and give a good indication on fuel quality.


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Notes: Some methods are described as “homebrew” methods. This means that these methods do not require a

chemical laboratory or highly skilled personnel to perform the tests. However, by performing these tests,

the producer can get a good indication on the quality of the biodiesel.

The following test methods have been set-up and are functional at the Marine Institute as of March 2013:

2, 3, 4, 5, 9, 10, 11, 12, 13, 15, 17

The following test methods will be installed at the Marine institute in the first quarter of year two: 1, 6, 7,

18

The following tests are suitable to be performed in a minimally equipped laboratory and moderate

personnel training requirements at the production site: 2, 4, 5, 9, 10, 11, 12, 13, 17

The remaining methods that have been indicated to be performed at MUN will be analyzed in

collaboration with laboratories of the CREAIT network.


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Table 8: Exemplary values obtained during oil and biodiesel (FAME) analysis of small-scale experiments in year one

Salmon Seal Specifications

Feedstock Raw Dried

Degummed and dried

Raw Dried Degummed and dried

FFA [%] 1.4 1.4 1.4 0.65 0.65 0.65

ideal: < 2%, acceptable: < 3%

water [ppm] 500 336 660 750 280 310 < 1500 ppm

Raw FAME (after MeOH removal)

soap (ppm) 2570 2605 2351 1639 1613 1182 low = higher yield

Washed FAME

Soap (ppm) after 2x wash 1900 623 1713 486 551 494 low = higher yield

Soap (ppm) when wash water is clear 55 18 18 33 15 12

< 20ppm (ASTM)

Yield after washing (ml) 1100 1100 1150 1000 960 1000

Yield (%] 91.67 91.67 95.83 83.33 80.00 84.75 > 80%

Water [ppm) 330

580 260 140

< 500 ppm

Ph 7 7 7 7 7 7 7

Flash point (°C)

> 93°C

Acid value 0.64 0.7 0.64 0.64 0.64 0.64 < 0.8 (ASTM)

Density (kg/mm3) 892 893 891

770-900 kg/mm

3

Kinematic viscosity (centistokes) 1.81 1.63 1.73

1,0-4,1 mm2/s


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5 Waste Generation and Management The proposed marine oil derived pilot scale biodiesel process does not result in a lot of waste. However,

depending on the process and feed stock used, it can produce some wastewater, resins, solids and

glycerol. The proposed biodiesel production processes includes three major operations: oil extraction,

pre-treatment, and biodiesel production (transesterification). Figure 11 shows the specific pollutants

generated from the different operations of biodiesel production.

5.1 Fish oil extraction This operation includes homogenization, heat oil extraction, centrifugation and oil polishing and

generates a major portion of solid waste (25% of the reaction volume) from the whole process.

However, solid waste generated from this operation is rich in protein content which can be utilized in

fish meal production and used as animal feed. 60% of the reaction volume (fish waste) results in a

wastewater stream which contains small amounts of protein. The protein can be recovered with flash

evaporation and the residual wastewater stream can be discharged into sewage.

5.2 Oil refining and pretreatment Fish oil refining or pretreatment includes degumming, hydration and filtration. The wastewater stream

generated after the degumming step is 2% in terms of reaction volume and contains phospholipids

(gum). Based on the concentration of the phospholipids, it can be directly discharged into the sewage

system or requires further treatment. Filter media and filtration residue come under the category of

hazardous waste as they contain impurities, oil and phospholipids. They should be disposed of under

hazardous waste rules and regulations.

5.3 Biodiesel production Biodiesel production includes transesterification, phase separation, methanol recovery, biodiesel

purification and ColdClear™ filtration. Glycerol is the major byproduct (waste) generated from the

transesterification reaction and requires proper handling and disposal strategies. The amount of glycerol

produced from biodiesel production can be quite significant and should not be overlooked.

Theoretically, the glycerol produced from biodiesel production is 20% of the volume of feedstock

processed, or in this case, 44 L if 220 L of salmon oil is processed for pilot scale biodiesel production.

Usually, the cost of glycerol disposal through a waste recycler (who then refines the material into fuels

and oils) is approximately $0.10 per liter of biodiesel produced. However, glycerol is becoming

increasingly sought after by pharmaceutical companies. These companies usually offer to cover the

costs of handling and transportation of the crude glycerol from the production facility to their site.

Therefore, the glycerol simply needs to be stored on-site (i.e. in recycled 200 liter drums at $5 each)

until they can be shipped off to the buyer. In this scenario, the cost of the drums is $0.0051 per liter of

biodiesel produced, a significant decrease from the cost that would be incurred if the glycerol were sent

to a recycler.

Amberlite™ BD10DRY™ polymer resin used in the dry wash column for biodiesel purification has the

ability to remove traces of soap, catalyst and glycerol. The resin needs to be disposed after 900-1600 kg


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of biodiesel treated (depends on the type of biodiesel). Spent polymer resin needs to be disposed under

hazardous waste regulations. Other filtration media and filtration residue waste generated at different

operations of biodiesel production need to be disposed according to hazardous waste rules.


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Figure 11: Specific pollutants generation during biodiesel production


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6 Next steps

6.1 Marine oil extraction As soon as the heat extraction equipment is installed, test runs will be performed on oil extraction from

salmonid waste streams as well as seal blubber. The obtained oil will be analyzed for its appearance and

smell, degree of oxidation, free fatty acid content as well as its fatty acid profile. The oil will be stabilized

and used for small scale or pilot scale experiments.

6.2 Completion of the pilot processing line A pre-treatment tank for performing the degumming step has to be procured and installed with the

existing pre-treatment tank. Specifications and dimensions of the pretreatment take will remain the

same as the existing pre-treatment tank with an additional high performance heater.

Until the installation is complete, preliminary pilot-scale experiments can be conducted using the

existing pre-treatment tank for both the degumming as well as glycerol stripping steps which will result

in an extended processing time. However, it is an acceptable intermediate solution.

6.3 Small scale experiments Further to the results in year one, additional small-scale experiments have to be performed on both

salmon and seal oil. Small scale experiments are divided into the following three parts:

6.3.1 Part A

Small scale studies on seal oil to identify the factors leading to the slow phase separation after

chemical transesterification

Once the factors have been identified, the recipe for transesterification will be optimized.

Parameters under investigation are: methanol/oil molar ratio, catalyst concentration, mixing

speed and reaction temperature.

6.3.2 Part B

Salmon oil pre-treatment and transesterification have to be further optimized. The most

important factors including methanol/oil molar ratio and degumming have been identified in

year one. The reaction conditions have not been optimized until now, however this is necessary

to ensure a safe, efficient and economic pilot scale process.

6.3.3 Part C

Once salmon and seal oil are extracted using the pilot line, the recipes developed with

purchased salmon and seal oil have to be confirmed on a small scale and adjusted if necessary

before the scale up can be performed.

6.4 Pilot scale experiments Pilot scale biodiesel production will be carried out using the same small scale optimized recipes after

CASD has achieved complete operational conditions of the pilot scale processing line and a sufficient

amount of extracted and characterized feedstock oils will be produced.


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It is expected that a constant biodiesel quality will only be achieved after 4-5 batches of operation due

to the glycerol pre-treatment steps and potential residues in the processing line. Therefore, pilot scale

production will only begin once enough feedstock oil is available for processing several batches in

sequence.

6.5 Biodiesel characterization In year one, an analytical test monograph has been developed that allows a good qualitative statement

on the biofuel quality without the need to use an accredited laboratory (refer to Table 7 for details).

These newly acquired testing capabilities at CASD will allow a better evaluation of small-scale

experiments and improve the validity of the results.

6.6 Development of work instructions While performing pilot scale studies, CASD will develop detailed work and sanitation instructions as well

as test procedures and a batch record system in order to initiate a solid quality control framework which

can be transferred to the client at the commercialization stage.

6.7 Development of a blending system Once biodiesel is produced on a pilot scale, studies on optimal blending of biodiesel with regular diesel

fuel will be performed. Preliminary studies will make evident whether or not a specialized blending tank

system is required or if the simple “pour and blend” technique is sufficient. Blending work instructions

will be developed for the client as blending will be performed on site.

6.8 Glycerol management Glycerol management has to be addressed during the process design phase as considerable amounts are

produced during transesterification and the cost for its disposal or re-use are significant. Year two will

include a review of the possible re-use and disposal options available and an initial financial statement

on expected costs.

6.9 Environmental analysis Waste streams produced during oil extraction and transesterification will be collected and analyzed.

Solids (Protein/bones): The sludge obtained during oil extraction will be dried and analyzed for its

composition and nutritional value as animal feed.

Stickwater: The stick water (water phase) obtained from the decanter centrifuge will be collected and

analyzed for its composition. Possible treatment options may include flash evaporation, in which the

water is removed and the remaining solids are added to the animal feed stream.

Washwater from degumming step: Wash water (containing phospholipids) generated from the

degumming step will be characterized for its pollution potential and environmental hazards.


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APPENDIX I: Description of Experiments

Appendix I

Description of Experiments


Contents

Experiment 1: Determination of acid value of oil ..................................................................................... 1

Experiment 2: Optimization of the solvent to oil molar ratio for chemical transesterification ............... 2

Experiment 3: Methanol removal on small scale ..................................................................................... 4

Experiment 4: Pre-treatment of oil with glycerol ..................................................................................... 5

Experiment 5: Effect of water degumming on phase separation ............................................................. 7


Page 1 of 11

Experiment 1: Determination of acid value of oil

Aim

The aim was to determine the amount of Free Fatty acids (FFA) present in the feedstock oils. If they

exceed their allowable limits, it is essential to reduce them prior to chemical transesterification.

Method

See Appendix II.

Results

Both oils have low FFA contents. According to literature, acid pre-treatment is recommended for FFA

between 2 and 5% and absolutely necessary above 5% prior to chemical transesterification.

Table 1: Results of Free Fatty Acid determination

mL NaOH FFA (%) Acid Number

Seal 0.85 0.65 1.3

Salmon 2.8 2.14 4.27

Discussion

Seal oil does not need pre-treatment to remove Free Fatty Acids as amount of FFA content in seal oil is

lower compared to allowable limits. The amount of FFA content in salmon oil is just above 2% and

pretreatment is not necessarily required. Literature suggests that fish oil transesterification is often

challenged by extremely high FFA values in the range of 10-20% which could not be seen here. The

relatively low of amount of FFA in salmon oil is considered to be due to the addition of antioxidant that

was added to the oil immediately after oil extraction.


Page 2 of 11

Experiment 2: Optimization of the solvent to oil molar ratio for

transesterification

Aim

The aim of this experiment was to investigate if a higher molar ratio can improve biodiesel and glycerol

phase separation and reaction speed of chemical transesterification.

Method

Four batches of feedstock oil transesterification were prepared. Table 2 outlines the parameters

involved in the chemical transesterification reaction. To different solvent to oil molar ratios were

compared side by side (6:1 and 12:1) while all other reaction parameters remains identical (amount of

catalyst used, reaction temperature, stirrer speed, and reaction time).

Table 2: Experimental parameters for the optimization of solvent to oil molar ratio

Batch # 1 2 3 4

Species Salmon Seal

Oil [mL] 500

MeOH [mL] 100 200 100 200

NaOH [g/L] 7.8 5.85

Molar ratio 6:1 12:1 6:1 12:1

Theoretically, 1 mol of alcohol is required to convert 1 mol of triacylglyceride. However, it has been

shown, that alcohol has to be added in excess to bring the reaction towards completion. Furthermore,

the reaction can happen in both directions and will find equilibrium depending on how much alcohol

and triglyceride are present. Different optima have been identified for different oil feedstocks. For

animal fats, several publications suggested that higher ratios (meaning a larger volume of Methoxide per

volume of oil) can lead towards better biodiesel and glycerol phase separation.

Molar Ratio Calculation

Average molecular weight of TAG: 962 g/mol (290 g/mol per fatty acid)

1000 [g] / 962 [g/mol] = 1.039 moles of oil

MW of Methanol (MeOH): 32.04 g/mol

32.04 [g/mol] * 1.039 [mol] * 6 = 199.74 g MeOH = Ratio 1:6 (~20 wt %)


Page 3 of 11

Results

For the batches using a 12:1 solvent to oil ratio, both reaction speed as well as phase separation were

much faster and the FAME (biodiesel) phase was much clearer. The results showed that a higher molar

ratio is needed to achieve optimal phase separation.

Figure 1: Phase separation after 24hrs. From left to right: Seal 6:1, seal 12:1, salmon 6:1, salmon 12:1

Discussion

This experiment was just a preliminary experiment and showed that further optimization of this

parameter is necessary. However, the produced FAME has to be analyzed for free and bound glycerol

content (Gas Chromatography) in order to make a qualitative statement about the optimal ratio for

transesterification of salmon and seal oil.


Page 4 of 11

Experiment 3: Methanol removal on small scale

Aim

During previous experiments in the pilot processing line, problems with methanol removal have been

encountered which resulted in lower density of the biodiesel and a low flash point (test fail). The

purpose of the experiment is to identify the root of the problem (oil or biodiesel properties). This

experiment is aimed at testing whether or not there is a problem with the oil or biodiesel which

prevents the methanol evaporation.

Method

The separated biodiesel phase after chemical transesterification was heated to 70°C with continuous

stirring until bubbling was observed (boiling methanol). The methanol removal was continued until no

more bubbling was detected and the density of FAME was evaluated with a hydrometer.

Results

No problem with methanol removal was observed. At 65°C bubbles started to rise up from solution and

continued to bubble for a few minutes until all methanol was evaporated. The density measurement

resulted in 895 and 897 g/L which are typical for marine biodiesel.

Discussion

The result indicated that the problem with methanol removal was not related to oil properties or

transesterification parameters. It is suggested that there are problems in the processing line, like

insufficient heating stirring or a blocked condensation unit or else which could all hinder complete

evaporation of the Methanol.


Page 5 of 11

Experiment 4: Pre-treatment of oil with glycerol

Aim

The aim was to observe differences in transesterification reaction if the oil is pre-treated with the

partially crude glycerol produced from the previous batch.

Method

The glycerol of a previous jar test was recovered and stored for two days. Then it was used to pre-treat

500 mL of oil prior to the transesterification reaction. The glycerol and new batch of salmon oil were

mixed for 1h at 50°C, followed by a settling phase of 1 hour and draining of glycerol.

1 L of seal oil was dried at 100°C for 1 h. 500 mL was then mixed with the glycerol obtained from a

previous batch. After the reaction time, the phases were allowed to settle out for 2 h before the

removal of the glycerol phase. Subsequently, two transesterification reactions were performed in

parallel:

Table 3: Experimental parameters, effect of pre-treatment with glycerol

Batch # 1 2

Species Seal

Oil [mL] 500

MeOH [mL] 200

NaOH [g/L] 4.35

Molar ratio 12:01

Pre-treatment Drying at 100°C for 1 h

Drying at 100°C for 1 h, followed by pre-treatment with glycerol

Results

Phase separation was faster for the batch that had been pre-treated with glycerol (batch 2). Figure 2

shows the oil feedstock for batch 1 and 2 before transesterification. Figure 3 displays the phase

separation over time for both batches. Figure 4 shows the appearance of the wash water during the

washing steps.

Discussion

A slightly faster phase separation was observed for batch 2, which had been pre-treated with glycerol.

During washing, batch 1 appeared to have lower soap content as phase separation was much quicker

and the wash water appeared clearer. More experiments on glycerol pre-treatment are necessary once

a series of transesterification reactions can be performed on pilot scale.


Page 6 of 11

Figure 2: Seal oil after drying and pre-treatment: Left: dried & glycerol treated (batch 2), right: dried only (batch 1)

Figure 3: Phase separation after transesterification at different time periods.

Left: dried & glycerol treated (batch 2), right: dried only (batch 1)

Figure 4: Appearance of wash water during washing steps. Left beaker: glycerol pre-treated, right beaker: dried only. A) 1

st

wash, water slowly poured, no agitation; B) 2nd

wash, water poured, no agitation; C) 3rd

wash, water poured, stirred for 20 s, phase separation after 10min; D) 3

rd wash, phase separation after 20 h; E) 4

th wash, water poured, stirred, phase separation

after 10min.


Page 7 of 11

Experiment 5: Effect of water degumming on phase separation

Aim

Aim of this experiment was to investigate if degumming of salmon oil by water treatment prior to

transesterification improves phase separation.

Method

Three batches were prepared in parallel. Table 4 outlines the experimental parameters for each batch.

Table 4: Experimental parameters for the optimization of the molar ratio

Batch # 1 2 3

Species Salmon

Oil [mL] 500

MeOH [mL] 200

NaOH [g/L] 7.8

Molar ratio 12:01

Pre-treatment

No pre-treatment

Drying at 100°C for 1h

3% water degumming, 1h, followed by drying

at 100°C for 1h

Before transesterification, the oil was analyzed for Free Fatty Acid content, and water content. After the

transesterification reaction, the phases were allowed to separate overnight before the glycerol was

removed. Subsequently, the methanol was boiled off at 70°C for 30min. The biodiesel phase was

washed with water until the wash water was clear (6 times). Unwashed biodiesel was tested for soap

content and the washed and dried biodiesel was analyzed for soap, content, acid value, pH, density and

kinematic viscosity.

Results

The quality of reaction and the reaction speed was judged by observing the color change after

Methoxide addition (fast color change indicates a faster glycerol formation, hence a faster reaction) and

also by the speed of phase separation.

For salmon oil transesterification, degumming clearly improved both reaction speed (Figure 5-Left) and

biodiesel and glycerol phase separation. After stirring was stopped, large glycerol droplets formed and

settled at the bottom quickly in batch 3 (degummed). The color of the mixture became dark almost

instantaneously (Figure 5 - Right) after the addition of methoxide, while batches 1 and 2 (no degumming

of oil) changed color slowly and never obtained a color as dark as batch 3 (Figure 6).

For seal oil transesterification, the degumming did not make a big difference in reaction speed and

phase separation. All three batches (4, 5, and 6) looked the same throughout the reaction. During

washing it was observed that phase separation of wash water and biodiesel was best for the degummed

and dried oil (batch 6). Also, the biodiesel obtained from degummed and dried oil had a more brownish

color compared to a more yellow color in batches 4 and 5. A yield difference was not observed.


Page 8 of 11

The free fatty acid content in both the salmon and seal feedstock were below the critical limit of 2%.

Hence, no treatment for removal of free fatty acids prior to transesterification was required. Also, the

water content was well below the critical limit of 1500 ppm. A good yield was achieved for salmon oil

(>90%), while the yield for the three seal oil batches was significantly lower (80-84%). Water washing

removed almost all soap residues and neutralized the pH. The kinematic viscosity and density were in

the normal range for Biodiesel.

The recorded values from the analytical tests performed on batches 1 to 6 are summarized in Table 5.

Figure 5: Comparison of the pretreatment on reaction speed during transesterification of salmon oil, batches 1-3 (left to right). The left picture shows the reaction 30s after Methoxide addition. The right picture shows the reaction right: 5 min

after Methoxide addition

Figure 6: Comparison of degumming on the color of the glycerol. Left: Salmon oil, no pre-treatment (batch 1), right: salmon

oil, degummed and dried before transesterification (batch 3)

Discussion

Degumming is definitely necessary for producing biodiesel from salmon oil as phase separation was

significantly faster and the obtained glycerol much darker which indicates a more complete reaction.

The quality of reaction and its completeness has to be further investigated by gas chromatography

which will yield information on the amount of free and bound glycerol.


Page 9 of 11

Table 5: Analysis of biodiesel, comparison of un-treated, dried and degummed and dried feedstocks

SALMON SEAL SPECIFICATIONS

Feedstock raw dried

degummed and dried

raw dried degummed and dried

Ffa [%] 1.4 1.4 1.4 0.65 0.65 0.65 ideal: < 2%,

acceptable: < 3%

Water [ppm] 500 336 660 750 280 310 < 1500ppm

Raw FAME (after meoh removal)

Soap [ppm] 2570 2605 2351 1639 1613 1182 low = higher yield

Washed FAME

Soap [ppm] after 2x wash 1900 623 1713 486 551 494 low = higher yield

Soap [ppm] when wash water is clear 55 18 18 33 15 12 < 20ppm (ASTM)

Yield after washing [ml] 1100 1100 1150 1000 960 1000

Yield [%] 91.67 91.67 95.83 83.33 80.00 84.75 > 80%

Water [ppm] 330 n/a 580 260 140 n/a < 500ppm

pH 7 7 7 7 7 7 7

Acid value 0.64 0.7 0.64 0.64 0.64 0.64 < 0.8 (ASTM)

Density [kg/mm3] 892 893 891 770-900 kg/mm

3

Kinematic viscosity [centistokes] 1.81 1.63 1.73 1,0-4,1 mm

2/s

APPENDIX II: Procedures for Oil and Biodiesel Analysis ad MI

APPENDIX II

Procedures


1 Contents 1. Oil Analysis ............................................................................................................................................ 3

1.1 Determination of the Free Fatty Acid (FFA) content in Oil and Biodiesel .................................... 3

1.2 Determination of Peroxide value of oil (AOCS, 1989) ................................................................... 4

1.3 Determination of p-anisidine value of oil (AOCS, 1989) ............................................................... 6

1.4 Visual inspection of oil and Biodiesel ............................................................................................ 7

1.5 Water content in Oil and biodiesel ............................................................................................... 7

1.6 Density determination in Oil or biodiesel ..................................................................................... 7

1.7 Viscosity determination in Oil or biodiesel ................................................................................... 8

1. Analysis of Biodiesel .............................................................................................................................. 9

1.8 PH determination in Biodiesel ...................................................................................................... 9

1.9 Completion of reaction ................................................................................................................. 9

1.10 Emulsification ................................................................................................................................ 9

1.11 Determination of the Soap Value in Biodiesel ............................................................................ 10

1.12 Corrosiveness to Copper ............................................................................................................. 11

1.13 Sulfated ash ................................................................................................................................. 11

1.14 Total and free glycerin ................................................................................................................ 12


Page 3 of 12

1. Oil Analysis

1.1 Determination of the Free Fatty Acid (FFA) content in Oil and Biodiesel Specifications: Fatty acid content Interpretation <2% Ideal for transesterification, no pre-treatment necessary 2-5% Pre-treatment recommended to remove FFA prior to transesterification >5% Pre-treatment necessary Acid number 0.10 mg KOH/g Specification for Biodiesel in Canada

A) Prepare a 0.025 N NaOH solution (0.1%)

a. Dissolve 1g NaOH in 1L distilled water

B) Sample preparation

a. Transfer 10ml Isopropyl alcohol into an Erlenmeyer flask

b. Add a few drops phenolphthalein

c. Tare

d. Add 1ml of oil/FAME. Record exact weight (~0.891g).

e. Titrate to the endpoint with 0.025N NaOH

C) Determination of amount of NaOH needed for transesterification

a. ml needed for titration + 5 (or 3.5) = amount of catalyst needed per L of oil

D) Calculation of %FFA

a.

v = volume of 0.025N NaOH needed to titrate sample

b = volume of 0.025N NaOH needed to titrate blank

N = Normality of NaOH solution

W = weight of SMP oil in grams

E) Determination of Acid number

a.


Page 4 of 12

1.2 Determination of Peroxide value of oil (AOCS, 1989)

Materials

Balance

Burette

Erlenmeyer flasks

Stop watch

Reagents

Chloroform

Formic acid

Glacial acetic acid

Starch (soluble)

Sodium thiosulfate solution (0.01 M)

Potassium iodide

Solutions

Potassium Iodide solution: Freshly prepare a saturated potassium iodide solution with deionized

water. Fill a flask to about 60% with potassium iodide then fill up with deionized water. The

solution must remain saturated (undissolved crystals must be present). The solution has to be

kept protected from light.

Starch solution: Dissolve 10 g starch in 1 L of deionized water with heating. Add 3ml formic acid

to stabilize the solution

Sodium thiosulfate solution. Prepare a 0.01 M solution with deionized water. The solution has to

be kept well sealed and in the dark. Determine the titer every week or prepare freshly.

Glacial acetic acid/chloroform mixture: Prepare a 3:2 mixture

Procedure

Transfer 3 g of oil sample to a 250 ml Erlenmeyer flask.

Add 50 ml of 3:2 acetic acid-chloroform

Swirl the flask to dissolve the sample.

Add 1.0 ml saturated potassium iodide solution and let stand for exactly 1 min

Add 100 ml of distilled water.

Add 1 ml of starch indicator and swirl flask to mix.

Titrate this mixture against 0.01 M sodium thiosulfate until the yellow color disappears.

Add 0.5 ml of starch indicator solution and continue titration with constant agitation till the

blue color disappears.

Test the samples against a blank (no oil added to mixture).

The peroxide value (milliequivalents (meq) peroxide/kg sample) can be calculated by the

following formula:


Page 5 of 12

V1 Volume of standard sodium thiosulfate used for titration of sample [ml]

V0 Volume of standard sodium thiosulfate used for titration of blank [ml]

N Normality of sodium thiosulfate solution

m weight of sample [g]

Interpretation

For crude fish oil, it is recommended that the peroxide value is 3-20 meq/kg. Higher values indicate

oxidation of the oil.


Page 6 of 12

1.3 Determination of p-anisidine value of oil (AOCS, 1989) Materials

25 ml volumetric flask

Test tube, 10 ml, with glass stopper

Pipettes, 1 ml and 5 ml

Spectrophotometer

Reagents

Para-Anisidine

2,2,4 Trimethylpentane (iso-octane), 99%

Glacial acetic acid, 99%

Solutions

Anisidine reagent, 0.25% w/v: Dissolve 0.25 g para-Anisidine in 100 ml acetic acid.

Place 0.5-4 g of oil accurately into a 25 ml volumetric flask.

Add 25 ml of iso-octane and swirl flask to dissolve the sample

Measure the absorbance of the fat solution against pure iso-octane at 350 nm in a 1 cm glass cell.

Pipette 5 ml of the fat solution into a test tube A and 5 ml iso-octane into a test tube B. Add 1 ml

anisidine reagent into test tubes A and B.

Seal the tubes with the stoppers, shake vigorously and leave in the dark for exactly 10 min.

Measure the absorbance of the content of tube A against tube B at 350 nm in a 1 cm glass cell.

Calculate the p-anisidine value as follows:

Ab Net absorbance of the oil solution after reaction with the p-anisidine reagent

Aa Absorbance of the oil solution

W Grams of sample

1.2 Correction factor

Interpretation

For crude fish oil, it is recommended that the p-anisidine value is 4-60. Higher values indicate advanced

oxidation of the oil.


Page 7 of 12

1.4 Visual inspection of oil and Biodiesel Samples

500ml of oil/biodiesel in a clear glass bottle

Reference sample of oil/biodiesel

Materials

15ml glass test tube

White sheet of paper

Light source

Procedure

Let the 500ml sample stand at room temperature for 48 hours. After that period, examine the

bottle for residues at the bottom or on the top without shaking.

Transfer a 15ml sample to the test tube and close the test tube with a lid

Hold the sample up into the light next to the reference sample and examine the color and

turbidity. Check for presence of small floating particles

Slowly invert the test tube. Observe for streaks during mixing, differences in viscosity, viscosity

and flow characteristics.

Note down all observations

Interpretation

Biodiesel and oil should be clear in appearance and not contain any residues. A cloudy appearance

indicates high water content. Residues settling out at the bottom of the flask indicate insufficient

filtering during the oil refining process. Streaks or various viscosities inside the test tube give an

indication of inhomogeneous oil.

1.5 Water content in Oil and biodiesel

Materials

Sandy Brae water test kit

Procedure

The procedure given by the manufacturer was followed (Appendix V)

1.6 Density determination in Oil or biodiesel Sample

200 ml of biodiesel in a graduated cylinder

Materials

Hydrometer


Page 8 of 12

Determine the specific gravity with a hydrometer at 60°F. The density of biodiesel is usually around 850

to 895 g/L. A lower viscosity may indicate residual Methanol in the biodiesel.

1.7 Viscosity determination in Oil or biodiesel

Sample

200 ml of biodiesel in a beaker

Materials

Viscometer

Determine the viscometer of the oil/biodiesel at 60°F.


Page 9 of 12

1. Analysis of Biodiesel

1.8 PH determination in Biodiesel

Measure the pH of the Biodiesel with a pH strip or a calibrated, standard laboratory pH meter.

Interpretation

The pH of biodiesel must be neutral (pH7), otherwise, the fuel will have corrosive characteristics and

damage the motor over time. If the pH is not 7, further washing is required to remove residual catalyst.

1.9 Completion of reaction Mix 25 ml Biodiesel with 225 ml Methanol in a 500 ml Erlenmeyer flask

Shake vigorously

Let the solution stand for 5 min

Interpretation

The biodiesel should be completely dissolved. If there is undissolved material at the bottom, this

indicates an incomplete reaction. Each ml of undissolved material corresponds to 4% by volume.

1.10 Emulsification Mix 50ml Biodiesel with 50ml H2O

Shake vigorously.

Interpretation

Biodiesel and water should separate within 30 min into two clear phases. If the water phase is cloudy or

if there is a soapy intermediate phase, this indicates the presence of residual soap and further washing

is required.


Page 10 of 12

1.11 Determination of the Soap Value in Biodiesel

Specifications

Soap Value Interpretation <41ppm Within ASTM standard <200ppm Should be ok for most engines 200-500ppm Need to wash more 2500ppm Value obtained for salmon raw fame (prior to

washing) on small scale

F) Check for residual catalyst

a. Place 250ml beaker on scale

b. Add 100ml isopropyl alcohol

c. Tare

d. Add 10g FAME

e. Add a few drops phenolphthalein

f. If liquid turns magenta, titrate to endpoint with 0.01N HCl

G) Check for Soaps

a. Add 20 drops Bromophenol Blue

b. Titrate to endpoint with 0.01N HCl

H) Calculation

a.

Result x 1,000,000 = [ppm]

B = ml of 0.01N HCl added during step B

C = Catalyst factor (320.56 for KOH and 304.4 for NaOH)

W = grams of FAME in solution (~10g)

I) Use results for Dry-washing

a. Soap value can be used to calculate the amount of Magnesol dry-wash resin needed

b.


Page 11 of 12

1.12 Corrosiveness to Copper

Sample

200 ml of biodiesel

Materials

Heating plate

Pot/container for oil that can be heated on heating plate

Polished copper strip: Copper strip must be polished immediately before use with steel wool or

400 grit sandpaper.

ASTM D130 standard color palette

Procedure

Method adapted from ASTM D130 and “Biodiesel – Basics and Beyond”

Heat oil and polished copper strip (fine steel wool or 400 grit sandpaper) to 50°C and cook for 3

hours.

After the reaction time, remove the copper strip from solution and compare the color of the

copper strip to ASTM standard color palette.

1.13 Sulfated ash

Sample

80 ml of biodiesel

Materials

Large crucible

Bunsen burner

Fume hood

Muffle furnace

Reagents

Concentrated sulfuric acid

Procedure

Place a SMP of Biodiesel (<80ml) in a large crucible (record weight of empty crucible) and slowly

heat until the fuel can be ignited.

Burn the fuel until no more flames or smoke appears.

Remove from flame and let cool to RT.

Add concentrated sulfuric acid until all residues are moistened. Slowly heat to evaporate acid.

Let cool to RT.

Add 3 drops of 50% sulfuric acid to moisten all residue.

Heat slowly to evaporate acid.

Place the crucible in furnace at 775°C for 30 min +/- 5 min. Determine weight of residue.


Page 12 of 12

Interpretation

Limits: max. 0.02% or 200ppm

1.14 Total and free glycerin Determined by Gas Chromatography according to ASTM D8495 (Appendix IV)

Appendix III

Water Test Kit Instructions

Appendix IV

Pilot Line Specifications

BDL-55-PPSX

Each BDL-55 Packaged Processor System "Patent Pending". Includes

• One size 3 carbon steel WVO pre-filter canister with a 400 micron polypropylene felt bag filter.

• One 84 gallon cone bottom 14 ga 304L stainless steel pretreat/phase one reaction vessel, with removable man-way cover for routine maintenance and cleaning of reactor. The reactor is equipped with integral sight gauge for visual indication of tank liquid level. The reactor vessel is insulated with 1/4" high density ceramic fiber insulation and wrapped with a hammered finish aluminum cover.

• One internal 220VAC, 1 Ph 2250 Watt, 1 Ph 316L stainless steel alloy heater assembly.

• One size 3 carbon steel WVO pre-filter canister with a 200 micron polypropylene felt bag filter.

• One 84 gallon cone bottom 14 ga 304L stainless steel phase two reaction vessel, with removable man-way cover for routine maintenance and cleaning of reactor. The reactor is equipped with integral sight gauge for visual indication of tank liquid level. The reactor vessel is insulated with 1/4" high density ceramic fiber insulation and wrapped with a hammered finish aluminum cover.

• One internal 220VAC, 1 Ph 2250 Watt, 1 Ph 316L stainless steel alloy heater assembly.

• One 84 gallon carbon steel holding/storage tank, equipped with integral sight gauge for visual indication of tank liquid level, the tank is painted with one coat of high particulate primer and two coats of polyurethane enamel paint.

• One integrated schedule 10 TIG welded carbon steel dry wash cell, equipped with 150 lb class access flanges for filling the cells and removal of spent media. The cell is sized to have 4 times the capacity of the dry wash media provided with the unit on delivery to allow for normal expansion of the media material over its usable life-span.

• One size 3 carbon steel final filter canister with a 5 micron polypropylene felt bag filter.

• One 15 gallon HDPE methyl/oxide mixing tank equipped with a 3/16” 304L stainless steel top with one mounted 220VAC, 1/3 hp, 1 Ph, TEFC high speed methyl/oxide prop mixer with stainless steel shaft and one SS 4” 3-blade prop.

• One pneumatic methanol/ methyl/oxide transfer pump. • Two 220VAC, ½ hp, 1 Ph, TEFC mixing pumps• Two 220VAC, ½ hp, 1 Ph, TEFC glycerin removal pumps• One 220VAC, ½ hp, 1 Ph, TEFC dry wash pump. • One 220VAC, ½ hp, 1 Ph, TEFC oil transfer pump. • One 220VAC, ½ hp, 1 Ph, TEFC fuel transfer pump.• One 220VAC, 2250 Watt, 1 Ph SS alloy flash separator/heater assembly.• One 304L methanol vapor recovery column/pot.

• One water cooled methanol recovery condenser, • All piping exposed internally to oil or biodiesel is carbon steel piping with the

exception of valves, access fittings and pumps. • All piping exposed internally to methyl/oxide is either FEP, PTFE or TYGON

high temperature tubing with the exception of valves, access fittings and pumps. • Two solid state low liquid level controllers specifically designed for direct

immersion in oil. • One optical flow safety switch for the methanol recovery flash separator. • One optical pump level control switch for the transfer cycle.• One optical pump level control switch for the dry wash cycle.• One NEMA 4/12 control panel, the control panel features digital temperature

controllers with illuminated oil-tight push buttons for each process stage. • All component/device wiring is enclosed in liquid tight flexible conduit with the

exception of pump motors designed to use SVO cord. • Two spring loaded 1.5 psig vacuum breakers. • Two spring loaded 1.5 psig vent valves. • All components of the packaged processor system are mounted on a MIG welded

heavy duty steel stand consisting of a base constructed from 6CX8 channel on edge with four 1” X ¼” square tubing vertical supports to a top 1” X ¼” frame with lifting lugs capable of supporting 2000 lbs combined weight. The stand is painted with one coat of high particulate primer and two coats of polyurethane enamel paint.

• The packaged processor system comes pre-charged with 88 lb Amberlyte BD10 Dry wash media to process a minimum of 10,000 gallon of biodiesel before recharging is necessary. The unit is accompanied with MSDS for the dry wash media.

• Step-by-step directions for general operation are shipped with the unit.• Biodiesel Logic Inc provides full documentation of factory operational testing and

also provides two full days of operational training at our factory for the owners operation personell prior to shipping.

Optional cold weather kits can also be installed to help optimize production during the cold winter months for an additional fee of $1,500.00 this option is recommended for all units in the northern half of the continental United States.

• Dimensions: 98"w x 26"d x 84"h • Weight: aprox 1300 Lbs. (empty)• Capacity: 55 gallons @ 8 hrs• Electrical: 110/220 VAC, 1 Ph, 60 Hz, 30Amp Circuit

Copyright 2009 Biodiesel Logic All Rights Reserved

Appendix V

Forms for long term biodiesel performance

testing

“Biodiesel production from fish oil”

Data Collection Sheet for KUBOTA diesel generator

(Sugarloaf site)

Week from _______________ to _________________

Step Mon Tue Wed Thu Fri Sat Sun

1 Time

2 Operator

3 Hours operated

4 total kWh used

5 Fuel consumed

6 Retrieve temperature data (USB stick)

yes

7 Retrieve power meter data (USB stick)

yes

8 Re-fuelled? yes yes yes yes yes yes yes

9 Re-fuelling form attached? yes yes yes yes yes yes yes

Key contacts: Marine Institute: Julia Pohling ([email protected], 709-757-4511), Wade Murphy ([email protected], 709-737-4534) Cooke Aquaculture: Wayne Piercey ([email protected], 709-xxx-xxxx)

mailto:[email protected]

mailto:[email protected]

Project contact at the Marine Institute: Julia Pohling (709-757-4511, [email protected]) Wade Murphy (709-737-4534, [email protected])

Long-term test: B5-B20 Biodiesel produced from salmon oil

Maintenance/Repair Record Sheet

for KUBOTA

Note: Please fill out this form for every maintenance and every repair that is performed on the Generator during testing with either petrodiesel or a biodiesel blend. You can add additional observations on fuel and engine

performance, residue formation etc. to the description.

Scheduled: ☐ Unscheduled: ☐

Date of maintenance/repair: ____ / ____ / _________

Date removed from service: ____________________

Date returned to service: ____________________

Maintenance/repair performed by: ____________________

Maintenance/Repair Description (detailed):

_____________________________________________________________________________________

_____________________________________________________________________________________

_____________________________________________________________________________________

_____________________________________________________________________________________

Hours of Labor: ____________________

Parts replaced: ____________________

Parts cost: ____________________


Long-term test: B5-B20 Biodiesel produced from salmon oil

Blending Record Sheet for KUBOTA diesel generator

Batch # _____

Date: ____ / _____ / _________

Operator: ______________________

Kind of diesel used: ________________________

(car, furnace, marked or

unmarked)

Volume of diesel added: ________________________ Liter

Volume of Biodiesel added: ________________________

Name of Biodiesel added (label): ________________________


“Biodiesel production from fish oil”

Re-fuelling Record Sheet for KUBOTA diesel generator

Date: ____ / _____ / _________

Operator: ______________________

Result of tank inspection: look into tank, screen for residue, especially at the bottom of the tank (color, amount, texture), remove all residue and make

sure the tank is dry (= free of water) before refueling. Indicate if you have removed any residues or water

_____________________________________________________________________________________

_____________________________________________________________________________________

_____________________________________________________________________________________

_____________________________________________________________________________________

Volume of fuel added: ________________________ Liter

Name of fuel (label): ________________________

Documents

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