PROXIMATE ANALYSIS AND LIGNIN

CONTENT OF HARDCROSS HARDWOOD

SAWDUST
(CUPRESSUS NOOTKATENSIS)

By

JOSHUA, JESSE AKOGU

(G2013/MENG/CHE/FT/261)

A THESIS SUBMITTED TO

THE DEPARTMENT OF CHEMICAL ENGINEERING

THE GRADUATE SCHOOL OF ENGINEERING
COLLEGE OF GRADUATE STUDIES
UNIVERSITY OF PORT HARCOURT

IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE AWARD OF THE

DEGREE OF MASTER OF ENGINEERING
IN CHEMICAL ENGINEERING

JANUARY 2016

i
 CERTIFICATION

UNIVERSITY OF PORT HARCOURT

COLLEGE OF GRADUATE STUDIES

PROXIMATE ANALYSIS AND KLASON LIGNIN CONTENT OF HARDCROSS

HARDWOOD (CUPRESSUS NOOTKATENSIS)

BY

JOSHUA JESSE AKOGU

G2013/M.ENG./CHE/FT/261
DEPARTMENT OF CHEMICAL ENGINEERING
FACULTY OF PROCESS AND ENERGY SYSTEM ENGINEERING

THE BOARD OF EXAMINERS DECLARED AS FOLLOWS:

THAT THIS IS THE ORIGINAL WORK OF THE CANDIDATE.THAT THIS THESIS IS
ACCEPTED IN PARTIAL FULFILLMENT OF THE REQUIREMENT FOR THE AWARD
OF MASTER OF ENGINEERING (M.ENG.) DEGREE IN CHEMICAL ENGINEERING.

NAME SIGNATURE DATE

PROF. AYOADE KUYE ……………………….. ………………………
SUPERVISOR

DR. ENVBUOWAN BENSON ……………………….. ……………………. ..

HEAD OF DEPARTMENT

PROF. O. D. ONUKWULI ……………………….. ……………………...

EXTERNAL EXAMINER

PROF. ADEWALE DOSUNMU ……………………….. ………………………

CHAIRMAN, BOARD OF EXAMINERS

ii
DEDICATION

To
My Parents

iii
 ACKNOWLEDGEMENT

I thank God for the strength, poise and resilience he blessed me with through this work.

I also wish to extend my gratitude to my supervisor, Prof. Ayoade Kuye, for his patience,

guidance, accessibility, simplicity and encouragement. These virtues have all helped me greatly.

Also worthy of note, is his drive for exceptional excellence that has spurred me on all through

this research and even in my dealings.

I also wish to extend my gratitude to the Petroleum Trust Development Fund (PTDF) for funding

this research work.

My gratitude also goes to my father, Mr. Joshua Akogu Akpochi and my mother, Mrs. Olive

Ojodunwene Akpochi, for their support and prayers. I also appreciate the concerns of my

siblings, Dr. Mrs. Ahuva Eleojo Nwokeoji, Esther O. Joshua, Joel E. Joshua, Joshua O. Joshua

and Zadok A. Joshua. I also wish to extend my gratitude to Dr. Obinna A. Nwokeoji for his

constant concern and advise.

I also commend the efforts made, advises rendered and concerns shared by my colleagues and

research mates; Olalekan Ogunwole and Tariah Awolaye. I am also very grateful to Julius

Ahiekpor for his tremendous contributions and considerations.

I also extend my gratitude to the Head of Department, Dr. Envbuowan Benson, Prof.

Chukwuma, Dr. Ipeghan Otaraku, Dr. Uyigue Lucky, Dr. Koyejo Oduola and all staff of the

Department of Chemical engineering, University of Port Harcourt, for their immeasurable

contributions. Also worthy of gratitude is the Late. Prof. C.C. Opara, for his advises that have

brought me this far.

iv
ABSTRACT

The lignin content in woody biomass, the proximate components, its porosity and particle
density are important factors that could influence certain properties of the bio-oil harnessed
through pyrolysis of the biomass or through any gasification process for combustible volatiles.
The extent of lignin content influenced by the particle sizes of the biomass was studied. The
research work studied the proximate components of ash content, moisture content, volatile
matter, fixed carbon and organic content. The experimental data obtained were presented on a
percent basis. The experiment for Percent Ash Content (PAC), Percent Moisture Content (PMC)
and Percent Volatile Matter (PVM) were conducted at 550±5 oC for 4 hours, 105±5 oC for 60
minutes and 550±5 oC for 10 minutes respectively. The experimental data obtained showed a
PAC of 0.47±0.10wt%, a PMC of 9.65±0.13wt% and a PVM of 99.34±0.11wt%. The proximate
components of Percent Fixed Carbon (PFC) and Percent Organic Matter (POC) were obtained as
0.18±0.03wt% and 0.136±0.03wt% respectively. The research work also studied the porosity and
particle density of the biomass. It was found that the porosity of the biomass (presented here as
an average mean of the porosity over the varying particle sizes tested) revealed a value of
0.96808% and the particle density (presented here as an average mean of the particle density
over varying particle sizes tested) obtained at 0.2425 g/ml. A study was ultimately conducted for
lignin content and the experimental data obtained was according to a particle size range. A total
mean was obtained at 19.02wt%. It was observed that there was a decrease in lignin content as
the particle size decreased.

v
 TABLE OF CONTENTS

TITLE PAGE - - - - - - - - - i

CERTIFICATION - - - - - - - - - ii

DEDICATION - - - - - - - - - iii

ACKNOWLEDGEMENT - - - - - - - - iv

ABSTRACT - - - - - - - - - v

TABLE OF CONTENTS - - - - - - - - vi

LIST OF ABBREVIATION - - - - - - - - xi

LIST OF FIGURES - - - - - - - - - xii

LIST OF TABLES - - - - - - - - - xiii

CHAPTER ONE

INTRODUCTION - - - - - - - - - 1

1.1 Background of study - - - - - - - 1

1.2 Statement of problem - - - - - - - 3

1.3 Aim and Objectives of the study - - - - - - 3

1.4 Significance of the study - - - - - - - 4

1.5 Scope of study - - - - - - - - 5

CHAPTER TWO

LITERATURE REVIEW - - - - - - - 6

2.1 Introduction - - - - - - - - 6

vi
2.2 Major components of wood - - - - - - 7

2.2.1 Cellulose - - - - - - - - 7

2.2.2 Hemi-cellulose - - - - - - - 8

2.2.3 Lignin - - - - - - - - 8

2.2.4 Elemental composition - - - - - - 9

2.3 Physical properties of Woody Biomass - - - - - 10

2.3.1 Moisture content - - - - - - - 10

2.3.2 Particle size - - - - - - - - 11

2.3.3 Porosity - - - - - - - - 12

2.3.4 Density - - - - - - - - 12

2.3.5 Ash content - - - - - - - - 13

2.3.6 Heating Value - - - - - - - 13

2.3.7 Volatile Matter - - - - - - - 14

2.4 Acid Insoluble Residue (Klason Lignin) Determination - - - 15

2.4.1 Acid insoluble residue Method - - - - - 15

2.5 Condensation of Lignin - - - - - - - 16

2.6 Proximate Analysis Methods - - - - - - 17

2.6.1 Percent Ash Content (PAC) - - - - - 18

2.6.2 Heating value - - - - - - - 18

2.6.3 Percent Volatile Matter (PVM) - - - - - 18

2.6.4 Percent Moisture Content (PMC) - - - - - 19

2.6.5 Percent Fixed carbon (PFC) - - - - - - 19

2.6.6 Percent Organic Content (POC) - - - - - 19

vii
 2.6.7 Porosity - - - - - - - - 19

2.6.8 Density - - - - - - - - 20

CHAPTER THREE

MATERIAL AND METHODS - - - - - - - 21

3.1 Collection of Biomass sample - - - - - - 21

3.2 Equipments Used - - - - - - - - 21

3.2.1 Soxhlet Extraction apparatus - - - - - 21

3.2.2 Muffle Furnace - - - - - - - 23

3.2.3 Precision balances - - - - - - - 25

3.2.4 Electromagnetic Sieve Shaker - - - - - 25

3.3 Proximate Analysis Components - - - - - - 27

3.3.1 Percent Ash content - - - - - - - 27

3.3.2 Percent Moisture content - - - - - - 27

3.3.3 Percent Volatile matter - - - - - - 28

3.3.4 Percent Fixed carbon - - - - - - 28

3.3.5 Percent Organic Content - - - - - - 29

3.3.6 Porosity - - - - - - - - 29

3.3.7 Density - - - - - - - - 30

3.4 Determination of Extractives - - - - - - 31

3.4.1 Determination of Water Soluble Extractives - - - 31

3.4.2 Determination of Ethanol Soluble Extractives - - - 31

3.5 Oven Dry Weight (ODW) - - - - - - - 32

3.6 Sample Preparation, Sieve analysis and

viii
Acid Insoluble residue extraction - - - - - - - 33

3.6.1 Sample preparation - - - - - - 33

3.6.2 Sieve analysis - - - - - - - 34

3.6.3 Extraction of Klason lignin - - - - - 34

3.7 Histochemical Tests for Lignin - - - - - - 35

3.7.1 Toluidine Blue Test - - - - - - - 35

3.7.2 Phloroglucinol - HCl Test - - - - - - 35

CHAPTER FOUR

RESULTS AND DISCUSSION - - - - - - - 36

4.1 Results - - - - - - - - - 36

4.1.1 Proximate Analysis of Hardcross hardwood sawdust - - 36

4.1.2 Determination of Extractives - - - - - 43

4.2 Oven Dry weight - - - - - - - - 45

4.3 Sieve Analysis and Lignin quantification - - - - - 45

4.3.1 Sieve analysis of pre-treated sawdust sample - - - 45

4.3.2 Lignin content quantification - - - - - 47

4.4 Acid Insoluble residue (Klason lignin) histochemical test - - - 49

4.4.1 Toluidine Blue test - - - - - - - 49

4.4.2 Phloroglucino-HCl test - - - - - - 51

4.5 Discussion - - - - - - - - - 53

4.5.1 Sieve Analysis of Pre-Treated Sawdust sample - - - 53

4.5.2 Influence of Oven-Dried Weight on proximate components - - 55

4.5.3 Effects of Acid Hydrolysis on biomass - - - - 55

ix
 4.5.4 Influence of Particle size on Lignin content yield - - - 57

4.5.5 Influence of Extractives - - - - - - 57

4.5.6 Colour change during Histochemical tests for Lignin content - 57

CHAPTER FIVE

CONCLUSION AND RECOMMENDATION - - - - - 59

5.1 Conclusion - - - - - - - - - 59

5.2 Recommendation - - - - - - - - 59

5.3 Contribution to knowledge - - - - - - - 60

REFERENCES - - - - - - - - - 61

APPENDIX - - - - - - - - - 66

x
 LIST OF ABBREVIATIONS

ASTM: American Standard for Testing and Materials

g: Grams

HCl: Hydrochloric acid

HHV: High Heating Value

LHV: Low Heating Value

ml: Millilitre

NREL: National Renewable Energy Laboratory

PAC: Percent Ash Content

PD: Particle Density

PFC: Percent Fixed Carbon

PMC: Percent Moisture Content

POC: Percent Organic Content

PVM: Percent Volatile Matter

µm: Micrometer

xi
 LIST OF FIGURE

Figure 2.1 - Structural formula of a Cellulose unit

Figure 2.2 - Most common molecular motif of hemicellulose

Figure 2.3 - A small piece of Lignin polymer

Figure 2.4 - Effect of moisture content in the heating value

Figure 2.5 - Condensation of Lignin during Sulphuric acid hydrolysis

Figure 3.1 - Soxhlet extraction apparatus

Figure 3.2 - Muffle furnace

Figure 3.3 - Electromagnetic sieve shaker

Figure 4.1 - Porosity versus particle size

Figure 4.2 - Effect of particle size on particl density

Figure 4.3 - Showing the weight composition of the water and ethanol soluble

extractives in relation to the sample weight

Figure 4.4 - Particle size distribution of the pre-treated sawdust

Figure 4.5 - Weight of Acid Insoluble residue obtained versus particle sizes

Figure 4.6 - Colour change after toluidine blue histochemical test

Figure 4.7 - Red colour formation during the phloroglucinol-HCl histochemical test

Figure 4.8 - Showing the gradation inside the dilute acid during Acid hydrolysis

xii
 LIST OF TABLES

Table 2.1 - Elemental composition of various wood types

Table 3.1 - Precision balances

Table 4.1 - Experimental data for Proximate components

Table 4.2 - Experimental data for the Acid insoluble residue, porosity and particle

density

xiii
14
 CHAPTER ONE

INTRODUCTION

1.0 Background of study

According to the World Bank Group (2013), it is estimated that about 1.2 bilion people do not

have access to power. This figure accounts for about 20% of the World population. This

deficiency in energy has been termed as ‘Energy poverty’ by the group. It also reported that

remote populations, unhealthy cookstoves, inefficient utilities, perceived investment risk, wasted

energy and untapped renewable energy could be possible factors that encourage this energy

deficiency. Amongst these factors, untapped renewable energy presents itself as the most

predominant factor. Renewable energy encompasses the harnessing of the solar , tidal, wind and

biomass energy sources for energy generation purposes. Behind only coal and oil, biomass

stands as the third-largest energy resource in the world (Jaya et al., 2011).

The use of biomass for energy generation offers many advantages, the most important being a

renewable and sustainable energy feedstock. It can significantly reduce net carbon emissions,

making it a clean development mechanism for reducing greenhouse gas emissions that have

plagued Nigeria and the world since the discovery, exploration, production and use of fossil

fuels. Presently, around the globe, there is a significant interest in using biomass for energy

generation as energy generation from fossil fuel continues to raise environmental concerns.

Using biomass for energy generation can bring a lot of environmental benefits (EUROPA,

European Commission).

The cheapest biomass sources are the waste products from wood or agro-processing operations

(Jaya et al., 2011). Wood or ligno-cellulose is a complex polymer and contains different

1
components with different properties. These components include Cellulose, Hemicellulose and

Lignin. These components are also reported to be the potential energy compounds found in

biomass (Kittiphop and Tharapong, 2012). It has been reported, that biomass can be used as a

fuel as such or can be converted to other energy matter like bio-oil, bio-char and bio-gas after

undergoing conversion processes like pyrolysis or gasification (Dinesh et al., 2006).

Bio-oil is a renewable liquid fuel. It offers the an opportunity of versatility in applications.

According to Dinesh et al., (2006), bio-oil chemically contains major group of compounds and

has a complex mixture of formic acid, isoeugenol, guaiacols, catecols, vanilins,

furancarboxaldehydes, syringols, pyrones, acetic acid, water, and other caboxylic acids.

The exact chemical nature of bio-oil is dependent on the feedstock. The review by Dinesh et al.,

(2006), had classified compounds found in bio-oil into five; hydroxyaldehydes, hydroxyketones,

sugars, carboxylic acids and phenolic compounds. The phenolic compounds are present as

monomeric units and oligomers derived from the coniferyl and syringyl builing blocks of lignin.

Lignin which is the second largest natural occuring polymer after cellulose contributes to the

extent of bio-oil yield expected of varying woody biomass as the lignin content varies for

different woody biomass (Dinesh et al., 2006). As reported by Dinesh et al., (2006), bio-oil

yields from wood pyrolysis are in the range of 72 – 80 wt%, depending on the relative amount of

lignin in the biomass. Various analytical methods have been developed over the years to estimate

lignin content in biomass. These methods for the quantitative determination of lignin fall into

two basic groups. First, there exist analytical procedures that are used to remove all constituents

except lignin. Secondly, procedures have also been developed that oxidize the lignin polymer out

of the cell-wall matrix of the biomass. The most commonly used analytical procedures that are

used to remove all constituents except lignin result in an insoluble lignin residue after utilizing

2
sulphuric acid for hydrolysis of the cell-wall polysaccharides after the removal of potential

interfering substances. The Klason lignin (KL) or Acid Insoluble Residue (AIR) procedure is one

of such sulphuric acid-utilizing hydrolytic method that was developed for use with wood. An

advantage of the Klason Lignin procedure is that it can be easily adapted to hydrolysis schemes

to obtain values for total carbohydrates, total uronic acids, and neutral sugar composition of

forage samples, as well as estimating lignin content.

1.1 Statement of problem

The quality and yield of bio-oil and other biomass generated energy products like the biogas and

bio-char is dependent on the type of feedstock used. Recently, there has been a growing concern

on how this relationship affects the whole harnessing process, from costing, plant set-up and to

retailing (Dinesh et al., 2006). Consequently, the need to know the composition of the

lignocellulosic components in woody biomass is necessary.

1.2 Aim and Objectives of the study

This study is a response to the call on research into the weight composition of lignin content in

biomass like the hardcross hardwood specie and how some of its proximate components,

extractive components, porosity and particle density affect its fuel properties.

Therefore, the main aim was to perform an analysis and obtain results for some proximate

components, determine its porosity and its particle density. Furthermore, extracting and

quantifying the lignin content in the hardcross hardwood biomass.

The objectives were to;

I. Determine the percent ash content, percent moisture content, percent volatile

matter, percent fixed carbon and percent organic content of the hardcross

hardwood.

3
 II. Determine the particle density and porosity of the Hardwood specie.

III. Perform an extractives analysis of the biomass

IV. Analyze its particle size distribution.

V. Experimentally estimate the lignin content of the Hardcross hardwood specie.

1.3 Significance of study

This study is significant considering the role of energy. Aside fossil fuels, alternative sources of

energy that are without the ills of the fossils fuels have become more and more appreciated. They

are clean and renewable sources of energy.

As a nation develops, the demand for energy asymptotically increases likewise. This demand

arises from the increased use of energy by the populace for domestic purposes and the industry

for their manufacturing processes. Ironically, not all of these demands are met. This is largely

caused by the ills of fossil energy and the remote location of some users. However, there exist an

abundance of biomass matter over the earth waiting to be harnessed irrespective of location. And

that has proved to be a very efficient alternative energy source over the years. Additionally, the

knowledge of what type of biomass is to be harnessed for energy or a composite of more than

one biomass is a very necessary knowledge, as not all biomasses present the same lignocellulosic

composition.

This study was aimed at analyzing the Hardcross hardwood biomass for its lignocellulosic

content of lignin, which is amongst the major precursors for energy derivation in any biomass

whether as a burning fuel or for energy matter derivation.

1.4 Scope of study

1. The study is limited to the experimental analysis of Hardcross hardwood

4
2. The experiment was performed for the determination of lignin content in Hardcross

hardwood.

3. For the proximate analysis, the research is limited to only the determination of its Percent

Ash Content (PAC), Percent Moisture Content (PMC), Percent Volatile Matter (PVM),

Percent Fixed Carbon (PFC) and Percent Organic Content (POC).

4. The lignin content quantification is limited to Very coarse( ≥1mm),Coarse (≥710µm),

Fairly coarse (≥500µm) and Fine (≥300 µm) particle sizes.

5
 CHAPTER TWO

LITERATURE REVIEW

2.0 Introduction

Bio-oil refers to oil sourced from wood. It has a characteristic dark brown colour. It is organic

and comprise of highly oxygenated compounds. It can also be referred to as pyrolysis oils,

pyrolysis liquids, Bio crude oil, wood liquids, woody biomass distillates, and pyroligneous acid.

It contains many reactive species that are largely responsible for some uncommon attributes that

it possesses. Chemically, it is a complex mixture of water, acids, and alcohols amongst many

others.

Biomass is a renewable resource and it is evenly distributed over the surface of the Earth than

other finite energy sources. Agricultural products, in particular residues from paper mills, are the

most common biomass resources used for generating electricity and power, including industrial

heat and steam, as well as for a variety of bio-based products. Energy generation from biomass is

receiving great attention due to environmental considerations and the increasing demands of

energy worldwide (Nwakaire and Ohagwu, 2009).

Chemically, the nature of bio-oil is dependent on the feedstock. This nature encompasses the

varying constituents of the biomass which include; extractives, cellulose, lignin and hemi-

cellulose. The constituents to a very large extent determine the quality and quantity of bio-oil

harnessed. Cellulose, hemicellulose, and lignin, the principal components of biomass, have

different reaction kinetics. Hemicellulose is the most reactive constituent, followed by cellulose

and lignin the least. Each component contributes to the behavior to an extent proportional to its

weight percent contribution to the composition of the raw biomass[ CITATION Sad10 \l 1033 ].

6
2.1 Major components of wood

The composition of wood is an important parameter in determining to a very large extent its

energy value. Wood has been reported to consist of three major basic polymers, namely cellulose

(C6H10O5), lignin (C6H10O3) (O.CH3) and hemicelluloses (Tillman, 1978). Wood and other plant

biomass are essentially composites constructed from oxygen-containing organic polymers. Minor

extraneous materials which are mostly organic extractives (Starch, coloring matter etc.) and

inorganic minerals (Gums, Tannins etc.) are present in wood (Sam, 2003).

2.1.1 Cellulose

Cellulose has a chemical formula (C6H10O5) n. At room temperature and atmospheric pressure, it

3
has a white powdery appearance. It has a density of 1.5 g/cm . It decomposes at higher

temperatures. Cellulose is hydrophilic, has no taste and odorless. It is water-insoluble and also

insoluble in most organic solvents, and it is biodegradable. It is a combustible component of the

biomass.

Figure 2.1: Structural formula of a Cellulose unit

2.1.2 Hemicellulose

7
Hemicellulose has an amorphous, random structure and possesses very little strength.

Hemicelluloses are the most reactive major component of wood decomposing in the temperature

range 200 - 260oC. Due to their lack in form crystals, it possesses a very instable thermal

property. Hemicellulose decomposition under pyrolytic conditions is postulated to occur in two

steps. First, its polymer breaks down into water soluble fragments followed by its conversion to

monomeric units, and finally it decomposes to volatiles. It produces more gases and less tar

compared to cellulose, and no levoglucosan (Soltes and Elder, 1981).

OH O

OH O
O O

O O
CH OH

CH OH
Figure 2.2: Most common molecular motif of hemicellulose

2.1.3 Lignin

Lignin is a highly linked, amorphous and high molecular weight phenolic compound. Lignin

serves as a bond between the wood fibers and as a stiffening agent within them. It is the least

reactive component of biomass. It is most commonly derived from wood, and is an integral part

of the secondary cell walls of plants. It is insoluble in organic solvent like water and alcohol but

soluble in weak alkaline solutions. Furthermore, it is tasteless and fibrous. It is one of the most

abundant organic polymers on Earth, second only to cellulose. It has been known to constitute

30% of non-fossil organic carbon and a quarter to a third of the dry mass of wood [ CITATION

enw15 \l 1033 ].

8
Its composition varies from one wood specie to another. An example of its composition from an

aspen sample is 63.4% carbon, 5.9% hydrogen, 0.7% ash, and 30% oxygen, corresponding

approximately to the formula (C31H34O11)n. It also lacks a defined primary structure. Its most

commonly noted function is the support through strengthening of wood (xylem cells) in trees

(Davin and Lewis, 2005).

H2COH

CH2

CH2
CH2
OH
CH2O CH

OH

Figure 2.3: A small piece of lignin polymer[ CITATION enw151 \l 1033 ]

2.1.4 Elemental Composition

The elemental composition of wood varies from species to species, but it is approximately 50%

carbon, 42% oxygen, 6% hydrogen, 1% nitrogen, and 1% other elements (mainly calcium,

potassium, sodium, magnesium, iron, and manganese) by weight (Jean, 1996).

Table 2.1: Elemental composition of various wood types (Samuel, 2014)

Sawdust Carbon, C Hydrogen, Nitrogen, N Oxygen, O Sulphur, S

type H
Radiata Pine 44.8 5.90 0.10 46.2 -
Poplar 45.5 6.26 1.04 47.2 -

9
 Pine-spruce 48.7 6.20 - 45.1 -

Bamboo 46.9 5.85 0.21 47.2 0.02

Japanese 50.8 6.80 0.10 42.3 -

Larch
White Oak 50.3 6.40 0.30 43.0 -
Beech 44.3 6.27 0.11 48.75 -

Pine 50.5 6.40 0.09 43.0 -

2.2 Physical Properties of Woody biomass

2.2.1 Moisture Content

This property of woody biomass plays a large role in determining the most suitable energy

conversion process. The moisture content of any biomass is an important efficiency parameter

when biomass combustion is the main consideration. Its determination is also of essence in most

carbohydrate analytical procedures. Water is held in biomass in two ways - either as a free liquid

and vapour that is contained in the cell cavities, or as a molecule that is bound within the cell

walls. It has been reported, that the moisture content tends to vary widely with biomass species,

its age, genetic differences and geographical location[ CITATION Woo15 \l 1033 ].

Furthermore, the moisture content in wood depends on several climatic conditions. These

conditions include, time of year when harvesting takes place, the duration and method of storage.

The moisture content of biomass can either be measured on a wet or dry basis. The wet-basis

expresses the ratio of moisture mass to the total mass of the substance while the dry basis

expresses the ratio of the moisture mass to the mass of dry matter[ CITATION Woo15 \l 1033 ].

(TotalWeightOfWetWood −OvenDryWeight )
MoistureContent= ×100
OvenDryWeight (2.1)

10
Moisture content influences the net calorific heating value of the woody biomass. Figure 1.4

illustrates the effect of moisture content on heating value.

Figure 2.4: Effect of moisture content on the heating value

2.2.2 Particle Size

Certain research findings have reported the effects of particle size to bio-oil yields (Bridgwater,

2003; Kersten et al., 2005; Jung et al., 2008; Kuye et al., 2012). The particle size distribution is

one of the widely used methods of characterizing woody biomasses. According to Bridgewater,

the heating rate potential of sawdust is an important function of its sizes which in turn controls

the rates of primary pyrolysis reaction (Bridgwater and Peacocke, 2000). The Smaller the

particle size of sawdust, the better advantages of shorter diffusion path, faster heating rates and

larger surface area it proposes, which further enhance the potency of the pyrolytic reaction in

producing bio-oil. Consequently, larger particles generally require more time to heat by intra-

particle conduction. The particle size can be reduced by pulverizing or grinding. However, the

11
drying and grinding of the sawdust feedstock induces a corresponding increase in the production

costs of the bio-oil (Solantausta and Oasmaa, 2003) (Kuye and Okorie, 2014)

2.2.3 Porosity

The porosity of sawdust can be defined as a measure of the void volume of its grains, composed

majorly of inter-spaces among and intra-spaces within the particles. In other words, porosity is

the percentage of sawdust volume occupied by air and water that fills the voids in the particle

(Boman et al., 2004). For proper modelling of bio-oil yield, the porosity of the wood biomass is

needful. According to Horisawa, the porosity can also be determined by following the method

(Horisawa, 1999):

Sawdust with apparent volume of 100cm 3 and known weight (Ws in grams) was at first placed in

a volumetric cylinder. Tap water was then poured gently into it until the surface of water reached

a marked line at 100cm3 level. A meshed top as a stopper was equipped at the 100cm 3 level, so

that the sawdust mostly floating on water would not go beyond its surface.

2.2.4 Density

This property depends largely on moisture content and porosity. It has been estimated that

cellulose has the highest density amongst the woody biomass constituents ρcellulose =1550 kg/m3,

but very soft wood (balsa wood) only has 50 kg/m3 (Isidoro, 2014). Tropical woods vary more

widely with a dry density as high as 1040 kg/m 3. The density of the wood cell wall is 1450 to

1550 kg/m3 (Weatherwax and Tarkow, 1987).

2.2.5 Ash Content

12
Ash is generally accepted to be the residue left after a material has been combusted. It has no

energy value and it is made up of the inorganic elements in the biomass. High ash-contents can

cause problems in many thermo-chemical processes[ CITATION Cel15 \l 1033 ]. The ash content of

plant species vary greatly, and is generally higher in agricultural residues. The ash present in

plants will depend on their stage of growth, the time of year, and their location.

The ash content tells us the amount of solid wastes present after complete burning process of the

fuel. It can be expressed as weight percent of the dry base or as a weight percent of the material.

It is reported that hardwoods have a higher ash content compared to softwoods. The higher the

ash content the lesser the heating value of the wood. Amongst wood biomass, this property

ranges between 0.08 to 2.3 %.

2.2.6 Heating Value

The heating value of any biomass is divided into two categories; the High Heating Value (HHV)

which is the heat released by combustion of a fuel at 25°C and returned to 25°C and the Low

heating value which is the heat released by combustion of a fuel at 25°C and returned to 150°C.

It is mathematically represented as;

LHV =HHV −LAtentHeatOfVaporisation

(2.2)

The higher heating value (HHV) of woody biomass is independent of moisture content and relies

on the chemical composition of the material. Graphically, a linear relationship exists between the

carbon content and its heat of combustion. Oxygen, nitrogen, and inorganic elements tend to

reduce this property. The Lower Heating Value (LHV) is perhaps more relevant than the HHV in

13
practical operations. It considers the energy required to vaporise the moisture of the biomass and

also that necessary to vaporise the water generated when the hydrogen and oxygen elements of

the biomass combine[ CITATION Cel15 \l 1033 ].

In addition, the Indian institute of Technology in Bombay, collected data on over 200 species of

biomass and fitted the following equation to the data[ CITATION Woo07 \l 1033 ]:

( KJ )
HHV =0 . 3491C +1. 1783 H−0 . 1034 O−0 . 0211 A +0 . 1005 S−0 . 015 N
(g ) (2.3)

Where C, H, O, A, S, N are the elemental composition of the elements Carbon, Hydrogen,

Oxygen, Sulphur and Nitrogen present in the wood specie.

2.2.7 Volatile Matter

The combustibles of solid fuels can be shared into two groups; Volatile matters and components

combusting as solid carbon. The share of volatile matter wood is typically high, 80 % of the

energy originating in the combustion of volatile matters.

2.3 Acid Insoluble Residue (Klason Lignin) determination

2.3.1 Acid-Insoluble residue method

Lignin is mostly insoluble in mineral acids, unlike the other cell wall components of biomass.

For this reason, lignin can be analyzed gravimetrically after hydrolyzing the cellulose and hemi-

cellulose fractions with sulfuric acid.

The Acid-Insoluble residue method was originally developed by Klason in the 1900s (Ronald

and Romualdo, 2005). It involves the use of 64 – 72% sulphuric acid to dissolve all the

carbohydrates in the wood sample leaving lignin as the insoluble residue.

14
The acid-insoluble residue content is used in conjunction with other assays to determine the total

composition of biomass samples. The results of acid-insoluble residue analysis are affected by

the incomplete hydrolysis of biomass. The results will be highly biased unless the sample is

hydrolyzed completely. Take care to mix the acid-biomass slurry thoroughly during the

concentrated acid hydrolysis. The results of acid-insoluble residue analysis are affected by the

timing of the acid digestion steps. The insoluble residue will dissolve slowly into solution in an

irreproducible fashion. The timing within this test method must be followed closely [ CITATION

AST09 \l 1033 ].

The Acid-Insoluble residue method involves; The weighing of 1-g test specimens in tared glass-

stoppered weighing bottles. Thereafter, they are dried in an oven for 2 hours at 100 to 105°C,

replace and stopper, and cool in a desiccator. Loosen the stopper to equalize the pressure and

weigh. Continue the drying for 1-hour periods until the weight is constant. Calculate the

percentage of moisture-free wood.

Air-dried wood sawdust is then placed in a glass stoppered weighing bottle or a small beaker

with a glass cover and 15 ml of cold (12 to 15°C) H 2 SO4 (72 %) is added. Mix the specimen

well with the acid by stirring constantly for at least 1 min. Allow to stand for 2 h, with frequent

stirring, at a temperature of 18 to 20°C. Wash the material into a 1-litre beaker or Erlenmeyer

flask, dilute to a 3 % concentration of H 2 SO4 by adding 560 ml of distilled water, and boil for 4

hours, either under a reflux condenser or in the nearly constant volume condition maintained by

the occasional addition of hot water to the flask.

After allowing the insoluble material to settle, filter into a filtering crucible that has been dried at

100 to 105°C and weighed in a glass-stoppered weighing bottle. Wash the residue free of acid

with 500 ml of hot water and dry the crucible and contents in an oven for 2 hours at 100 to

15
105°C. Place in the weighing bottle, cool, loosen the stopper of the bottle, and weigh the contents

of the crucible as lignin. Repeat the drying and weighing until the weight is constant.

2.4 Condensation of Lignin

The condensation of Lignin in the presence of Sulphuric acid is believed to involve the

condensation of reactive groups which are at least partly of benzyl alcoholic character. During

this condensation, the carbon atoms of some of the elements become attached to two aromatic

nuclei with the loss of water.

R’

R’
R’
H2SO4
OCH3 H-C
H – C – OH H OCH3 + H2O

OH
H (OCH3)

CH3O H (OCH3)

O_ _ _

O_ _ _

Figure 2.5: Condensation of Lignin during Sulphuric acid hydrolysis

Furthermore, the carbohydrates been hydrolyzed during Acid hydrolysis are classified as

polysaccharides and can be grouped into three fractions; Cellulose which is the most resistant to

chemical disruption, Hemicellulose which can be extracted by relatively strong alkali solution or

mild acid hydrolysis (Dey and Brinson, 1984).

2.5 Proximate Analysis Methods

These methods will determine the composition of the biomass fuel in terms of gross components.

The value of proximate analysis is that it identifies the fuel level of the as-received biomass

16
material. In addition, Proximate analysis is a standardized procedure that gives an idea of the

bulk components that make up a fuel, it is done to determine the Percent Volatile Matter (PVM)

content, Percentage Ash Content (PAC), Percent Moisture Content (PMC), Percent Fixed Carbon

(PFC), Percent Organic Content (POC), Porosity and Particle Density (PD) of the biomass.

These properties were determined in accordance with ASTM analytical methods.

2.5.1 Percent Ash content (PAC)

The method seeks to estimate the ash content of the biomass sample. The amount of inorganic

material in biomass, either structural or extractable, should be measured as part of the total

composition. Structural ash is inorganic material that is bound in the physical structure of the

biomass, while extractable ash is inorganic material that can be removed by washing or

extracting the material. Extractable ash can be the result of soil remaining in the biomass (Sluiter

et al., 2008). The ash content is an index that measures the mineral content and other inorganic

matter in biomass and is used in conjunction with other procedures to determine to total

composition of biomass samples.

The Percentage Ash Content (PAC) is determined by heating 2 g of the 425 microns sawdust

particle size in the furnace at a temperature of 550°C for 4 h and weighed after cooling to obtain

the weight of ash.

2.5.2 Heating Value

The Higher Heating Value (HHV) of a sample is reliant on the chemical composition of the

material. A linear relationship exists between the heat of combustion and the Carbon content of

the substrate while Oxygen, Nitrogen and Ash tend to reduce the value. This is the most

important fuel property.

17
2.5.3 Percent Volatile Matter (PVM)

The Percentage Volatile Matter (PVM) is determined by measuring 2 g of the 425 microns

particle size sawdust in a crucible and placing it in an oven until a constant weight is obtained.

The sawdust is then kept in a furnace at a temperature of 550°C for 10 min and weighed after

cooling in a dessicator. This component helps us know how much of the sawdust is combustible.

2.5.4 Percent Moisture Content (PMC)

The Percentage Moisture Content (PMC) is found by weighing 2 g of the sawdust (E) and oven

drying it at 105°C until the mass of the sample is constant. The change in weight (D) after 60 

min is then used to determine the PMC of the sawdust sample.

2.5.5 Percent Fixed Carbon (PFC)

The percentage fixed carbon (PFC) is computed by subtracting the sum of PVM, PAC and PMC

from 100;

¿ carbon=100 %−(PAC + PVM + PMC ) (2.4)

2.5.6 Percent Organic Content (POC)

Organic matter influences many of the physical, chemical and biological properties of biomass.

Some of the properties influenced by organic matter include biomass structure and shear

strength. In addition, this proximate property also affects the water holding capacity, biological

activity, and water and air infiltration rates.

2.5.7 Porosity

Porosity is calculated by the following formula;

Va
Porosity ( % )=( )Vo
×100 (2.5)

18
Where Va and Vo are the volume of poured water (cm 3) together with the water in the sawdust,

and the volume of sawdust respectively. The volume of poured water with the water in the

sawdust could be calculated by the formulae;

V a ( cm3 )=W comb−W s−W vs (2.6)

Where;

Wcomb = Combined weight (grams) of volumetric cylinder, sawdust particle and poured water

(grams)

Ws = Weight of sawdust particles (oven dry weight equivalent)

Wvs = Weight of volumetric cylinder

2.5.8 Density

There exist three categories of evaluating biomass density (John, 2009). These are;

True density, ρTrue ;

Total mass of biomass

ρTrue= (2.7)
Solid volume of biomass

The True density is also referred to as the Particle Density (PD).

Apparent density, ρ Apparent ;

Total mass of biomass

ρ Apparent = (2.8)
Volume of solids an d internal pores

Bulk density, ρ Bulk;

Total mass of biomass

ρ Bulk= (2.9)
Bulk volume occupied by particles

19
 CHAPTER THREE

MATERIALS AND METHODS

3.1 Collection of Sawdust sample

20
The sawdust sample was obtained from Dagogo timber sawmill located by the water bank of

Illoabuchi Street, Mile one, Diobu, Port Harcourt Nigeria. The sawdust sample used for analysis

was Hardcross hardwood (Cupressus nootkatensis). The sample was collected as described by

Okorie (2014) using the standard procedure ASTM D346/D346M-11 (2011). The study was

conducted at the Petroleum Trust Development Fund (PTDF) Bio-oil research laboratory, located

at the ETF Gas engineering building, Abuja campus, University of Port Harcourt, Port Harcourt,

Rivers state and stored under room conditions until its use.

3.2 Equipment used

3.2.1 Soxhlet Extraction Apparatus

The Soxhlet extractor (Pyrex Quickfit UK, EX5/63, BS2071, 24/29 fit) was 250ml in capacity.

Coupled with the extractor was a reflux condenser (Pyrex Quickfit UK, CX7/06/SC, 40/38 fit).

The apparatus also consisted of a Metallic stand and a Heating element (J.P. Selecta S.A,

3003145, S/N 0548400, 230V, 50/60 Hz, Spain). Figure 3.1 shows a complete set-up of the

Soxhlet extraction apparatus.

21
 Figure 3.1: Soxhlet Extraction apparatus

3.2.2 Muffle Furnace

22
Figure 3.2 represents a pictorial display of the Muffle furnace employed for the analysis. Drying

of sawdust sample before and after extraction procedures was achieved using a muffle furnace

(J.P. Selecta, S.A, 582543 S/N, 230 VAC, 00-C/2000367, 50/60 Hz, 3500W, Spain).

23
 Figure 3.2: Muffle Furnace

3.2.3 Precision balances

24
Two different scales with varying degree of precision were used for this study. Table 3.1

illustrates these balance types and their varying specifications.

Table 3.1: Precision balances

Precision Balance 1 Precision Balance 2

Manufacturer RADWAG Electronics RADWAG Electronics
Model WTB 200 WLC 6/A2
Serial Number 316283/11 379969/13
Maximum Load 200 grams 6 Kilograms
Minimum Load - 5 grams
Precision 0.001 grams 0.1 grams
Power:

Input 11 V/50 Hz 11 V AC

Output 230 V/50 Hz 230 V/50 Hz

Year of Manufacture 2011 2013
Country of Manufacture Poland Poland

3.2.4 Electromagnetic Sieve Shaker

To evaluate the Particle sizes of the sawdust sample before Lignin quantification, an

Electromagnetic Sieve shaker (BA 200N, 01006, 230 V, 50/60 Hz, 450 VA, Spain) was used.

Figure 3.3 depicts the Electromagnetic Sieve shaker used.

25
 Figure 3.3: Electromagnetic sieve shaker

3.3 Proximate Analysis Components

The Proximate Components of the sawdust sample were studied using the methods as described

by Joseph and others (2012).

3.3.1 Percent Ash Content (PAC)

26
To obtain the Ash content of the sample, a porcelain crucible was weighted and oven dried until

its weight was constant. A sample weight of 2g was then measured and placed in the crucible.

The crucible and content were then weighted to the nearest 0.001g. The sawdust sample was then

ashed at 550 degree Centigrade. Ashing was conducted for 4 hours at a regulated temperature

range of 550±5oC. After 4 hours, the crucible was carefully withdrawn, and cooled in a

dessicator. After cooling, it was weighted to the nearest 0.001g. This procedure was conducted in

triplicate. The Ash content was found on percentage basis. This was achieved using the

mathematical formulae;

Z
PAC= ×100
A (3.1)

Where,

Z = Weight of ash, in grams (g)

A= Weight of the oven dried sample at 105oC, in grams (g)

3.3.2 Percent Moisture Content (PMC)

To obtain the Percent Moisture content, two grams of the sawdust sample was placed in a

porcelain crucible. The crucible was weighted. It was then dried at 105 ± 5 degree Centigrade for

60 minutes. After 60 minutes, the crucible was withdrawn and cooled. The crucible was then

weighted to the nearest 0.001g. This procedure was conducted in triplicate. The moisture content

was found on a percentage basis using the formula as described in ASTM D1348−94 (2008);

D
PMC= ×100
E (3.2)

Where; D = Weight change, in grams (g)

27
 E = Original weight of sawdust, in grams (g)

3.3.3 Percent Volatile Matter (PVC)

A weight of 2g of sawdust sample was measured into the crucible and weighted to the nearest

0.001g. It was then ashed at a temperature range of 550 ±5oC for 10 minutes. Tthe crucible was

then withdrawn and cooled. After cooling, the crucible was then weighted to the nearest 0.001g.

This procedure was conducted in triplicate. This parameter was then calculated on a percent

basis using a formula as recommended by Joseph and Others (2012);

A−B
PVM= ×100
A

(3.3)

Where; A = Weight of the oven-dried sample at 105oC, in grams (g)

B = Weight of the sample after 10 min in the furnace at 550°C, in grams (g)

3.3.4 Percent Fixed Carbon

This proximate component was determined on a percentage basis using the formulae;

FixedCarbon=100 %−(PAC +PVM ) (3.4)

.3.5 Percent Organic Content

To determine the Percent Organic Content a standard as recommended by ASTM D2974 (2002)

was replicated. The Mass of an empty and dry porcelain dish was determined and recorded. An

oven dried sample of 1.982 grams was then placed insided the porcelain dish and the Mass was

recorded. The dish was then placed in a muffle furnace and allowed to heat for 4 hours at 550±5

degrees centigrade. The porcelain dish was then withdrawan from the furnace and allowed to

cool to room temperature. The mass of the dish containing the ashed sample is determined and

28
recorded. The Percent Organic Content was then determined using the following mathematical

expressions;

M O=M −M Ash
Sample

(3.5)

Where;

Msample is the mass of the Oven dried sample in grams

Mash is the mass of the ash in grams

Mo is the mass of the organic matter in grams

The Percent Organic Content (POC) was then determined using the mathematical expression;

Mo
OC= ×100
M Sample

(3.6)

3.3.6 Porosity

To determine the Porosity, then a sample with apparent volume of 100 cm 3 and known weight

(Ws) was placed in a volumetric cylinder. Water was then poured into it until the surface of

water reached a marked line at the 100 cm3 level (Rizki et al., 2010). Porosity was obtained

using the formula:

Va
Porosity(%)=
( )
Vo
×100
(3.7)

29
Where Va and Vo are the volume of poured water (cm 3) together with the water in the sawdust,

and the volume of sawdust respectively. The volume of poured water with the water in the

sawdust could be calculated by the formulae;

(3.8)

Where:

W comb = Combined weight of volumetric cylinder, sawdust particle and poured water, in grams

W s= Weight of sawdust particles (oven dry weight equivalent), in grams (g)

W vs= Weight of volumetric cylinder, in grams (g)

3.3.7 Particle Density

The Particle density was calculated by adding the sawdust (on the 100 cm 3 mark) into a

graduated volumetric cylinder to reach the marked 100 cm3 volume (Vo), and its weight (g) was

then known by subtracting the combined weight of sawdust and volumetric cylinder (Wsv) with

the weight of empty volumetric cylinder (Wv) (Rizki et al., 2010). The Particle density was then

calculated using the formulae:

( W sv −W v )
PD=
Vo (3.9)

Where;

W svis the combined weight of the sawdust and volumetric cylinder, in grams (g)

W v is the weight of the volumetric cylinder alone, in grams (g)

V o is the cubic basis tested usually around 100 cm3

3.4 Determination of Extractives

A weight percent calculation on the extractive content in the sawdust sample was performed on

the sawdust sample according to the standard method described in ASTM E1690 (2003) and

30
NREL 42619 (2008). The extraction was done and the extractives were dried at a temperature of

105 ± 5 degree Centigrade until a constant weight was observed.

3.4.1 Determination of Water soluble Extractives

The quantification for the water soluble extractives contained in sawdust sample was performed

according to the method described by NREL 42619 (2008). A paper tray was wiped and

weighted to the nearest 0.001g. It was then dried at 105 ± 5 degree Centigrade (o C) until it

assumed a constant weight. 50g of Hardcross sawdust was then placed in the paper tray and

weighted to the nearest 0.001g. The combined weight of the tray and the sawdust sample was

then recorded. The sample was charged into a porous thimble and then placed inside the Soxhlet

extractor. 500 ml of water was then dispensed into a 500 ± 50 ml round bottom flask, which was

then placed on a heating mantle. For controlled boiling, four (4) boiling chips of 0.03g weight

each were placed in the flask. The Soxhlet extraction apparatus was then coupled to the 500 ml

flask. And extraction was conducted for four hours. After the extraction, the supernatant was

collected. It was evaporated until it reached a volume of about 15 ± 5 ml. Then it was dried at

constant heating (105 ± 5 degree Centigrade) until a constant weight was reached.

3.4.2 Determination of Ethanol Soluble Extractives

To conduct this Compositional analysis, the sample was dried until there was no variance in

weight[ CITATION AST03 \l 1033 ]. The dried sample was then weighted and charged into a thimble

and then placed inside the Soxhlet extractor. 360 ml of water was then measured using a Pyrex

volumetric cylinder and dispensed into a 500 ml round bottom flask, which was then placed on

the heating mantle. For controlled boiling, four (4) boiling chips of 0.03g weight each were

placed in the round bottom flask. The Soxhlet extraction mechanism was then coupled to the

flask. After the extraction was conducted for four hours using ethanol, the supernatant was

31
collected. It was evaporated until it reached a volume of about 15 ± 5 ml. It was then dried at

constant heating (105 ± 5 degree Centigrade) until a constant weight was reached.

3.5 Oven Dry Weight (ODW)

For the proximate analysis, a basis of 2g of the sample was used. A porcelain crucible was wiped

and dried for 30 minutes. It was then weighted to the nearest 0.001g. The sample was then placed

in the crucible and the combined weight recorded. The crucible was then dried at a temperature

of 105 ± 5 degree Centigrade. The crucible was weighted at interval of 20 minutes until it was

observed that the weight was constant. The crucible and its contents were then weighted to the

nearest 0.001g. And then the Oven Dry Weight (ODW) was calculated using the expression as

described by Boone and Wengert (1998);

( W TS −W T )
ODW = ×% TotalSolids
100

(3.10)

Where;

W TS is the weight of crucible and sample, in grams (g)

W T is the weight of the crucible used, in grams (g)

%Total solids is calculated using the formulae;

( W TS −W T )
%TotalSolids= ×100
W Sr (3.11)

Where;

W TS is the weight of crucible and sample, in grams (g)

W T is the weight of the crucible used, in grams (g)

W S is the weight of the sample as received, in grams (g)

r

32
The Oven Dry Weight (ODW) can also be calculated using the method as recommended by

Boone R.S. and Wengert E.M (1998);

100×M Original
ODW=
MC+100 (3.12)

Where;

MOriginal is the Original mass of the sample in grams

MC is the Moisture Content expressed as a percent

3.6 Sample Preparation, Sieve Analysis and Acid Insoluble Residue (AIR) Extraction

3.6.1 Sample Preparation

All plant materials contain phenolic compounds that can potentially interfere with the Acid

Insoluble residue (Klason lignin) analysis. No single procedure exists for removing such

compounds prior to the lignin extraction. Even though the foregone procedures as described in

Section 3.4 represent the most widely used and accepted methods for removing extractives from

plant tissues prior to lignin analysis, it is worth noting that acetone-water (9:1) can be equally

effective as ethanol. However, Acetone-Water extraction can remove up to 10% of the lignin

from some woody tissues, while subsequent washing with hot water can remove a further 5-10%

[ CITATION Jef97 \l 1033 ]. It was for this reason that only the mild procedure of Water and Ethanol

extraction were adopted prior to Lignin extraction to minimize error.

33
3.6.2 Sieve Analysis

Sieve analysis was further conducted on the extractives – free sample. After extractives removal,

the sample was then sieved using the electromagnetic sieve shaker and the particle sizes ≥1 mm,

≥710 µm, ≥500 µm and ≥300 µm were then selected for the Lignin analysis.

3.6.3 Extraction of Klason Lignin

The Klason Lignin was extracted using a standard procedure as described by ASTM D1106-96

(2007). The extraction was done in triplicates for 310 micrometer, 500 micrometer, 700

micrometer and 1 millimeter particle sizes. One gram (1g) of the sample was the basis for each

extraction. 72 ml of concentrated Sulphuric acid (H2SO4) was contacted with 28 ml of water.

This was done to achieve a 72% mixture of hydro-sulphuric acid. The mixture was allowed to

cool to about 18 – 25oC. For each 1g of sample, 15 ml of 72% hydro-sulphuric acid was

contacted. It was then stirred vigorously for the next 1 – 2 minutes to avoid lumping. Thereafter,

it was left for 2 hours with intermittent agitation. After the 2 hours elapsed, the mixture was then

diluted to a 3% Sulphuric acid concentration. This was achieved by the addition of 560 ml of

distilled water. The mixture was then boiled for the next 4 hours with intermittent stirring. After

4 hours, it was observed that, aside from the insoluble solid precipitate at the bottom of the flask,

there was also an insoluble colloidal formation. The mixture was then left to settle. The acid-

water solvent was then decanted. The residue was then washed with 500 ml of hot water to

remove any other contacted acid. It was then emptied into a filter paper to drain. Thereafter, it

was dried at 105 ± 5 degree Centigrade for 2 hours. After drying, it was then emptied into a

porcelain crucible. The crucible was then weighted and the difference in weight between the

crucible and its contents was then found out as the weight of the Acid Insoluble Lignin. The

percent weight of Lignin was found using the formulae;

34
 PercentWeightOfLignin= ( W CL −W C )×100

(3.13)

Where; W CL is the weight of crucible and Lignin, in grams (g)

W C is the weight of the crucible used to measure, in grams (g)

This percent weight of Lignin was found in relation to the 1g of sample used for each analysis.

3.7 Histochemical Tests for Lignin

Analytical tests were carried out on the Acid-insoluble residue assumed to be Lignin. These tests

are chemical methods that helped to indicate the presence of lignin in the residue by staining.

They were colour based reactions and are widely used diagnostics test for Lignin.

3.7.1 Toluidine Blue Test

For the Toluidine blue test, 0.5 g of Toluidine blue was dissolved in 100ml of distilled water. 0.1

± 0.01g of the sample to be tested was soaked in ethanol for 5 – 10 minutes and then placed on a

white background paper. Using a dropper, 3 drops of the dye was introduced on the sample. A

colour change from blue to pink was then observed.

3.7.2 Phloroglucinol - HCl Test

To test for Lignin using the Phloroglucinol-HCl procedure, a 2% (w/v) solution of

Phloroglucinol was prepared in 95% ethanol, and stored in a sealed bottle. Immediately prior to

use, the residue was soaked in ethanol for 5 – 10 minutes and then placed on a white background

paper. Two parts Phloroglucinol solution was mixed with one part concentrated hydrochloric

acid (HCl), and applied to the residue. A red colour was observed which then fades almost

immediately.

35
 CHAPTER FOUR

RESULTS AND DISCUSSION

4.1 Results

The results obtained from this study are presented in Table 4.1 and Table 4.2. The proximate

components analysis results are presented in Table 4.1. The results for the acid insoluble lignin

quantification, extractive analysis, particle density and sieve analysis are presented in Table 4.2

4.1.1 Proximate Analysis of Hardcross hardwood sawdust

The proximate analysis for the sawdust sample used in this study is given in the Table 4.1. The

results indicated a Mean of 0.47±0.10, 9.65±0.13, 99.34±0.11, 0.18±0.03 and 0.136±0.03 for the

Percent Ash Content (PAC), Percent Moisture Content (PMC), Percent Volatile Matter (PVM),

Percent Fixed Carbon (PFC) and Percent Organic Content (POC) respectively. These results

represent the total mean of Nine (9) experimental runs for each proximate analysis component.

36
37
 Run, wt%
Proximate Mean, wt%
Components
1 2 3
PAC 0.32 0.38 0.35 0.58 0.55 0.56 0.50 0.49 0.52 0.47±0.10

PMC 9.82 9.48 9.77 9.75 9.53 9.56 9.78 9.64 9.56 9.65±0.13

PVM 99.53 99.43 99.47 99.19 99.25 99.27 99.35 99.31 99.32 99.34±0.11

PFC 0.15 0.19 0.18 0.23 0.20 0.17 0.15 0.20 0.16 0.18±0.03

POC 0.09 0.11 0.10 0.16 0.16 0.16 0.15 0.14 0.15 0.136±0.03

Table 4.1: Experimental data for Proximate components

38
 Table 4.2: Experimental data for the Acid insoluble residue, Porosity and Particle Density

S/N Particle Size Experiments Percent weight of Acid Porosity, % Particle Density, grams
Insoluble residue, %wt per per ml (g/ml)
gram

1 Very coarse 1 0.236 0.9020 0.2505

(≥1mm)
2 0.221 0.8948 0.2510
3 0.223 0.8913 0.2497
Mean 0.2267 0.8960 0.2504
2 Coarse 1 0.181 0.8598 0.2420
(≥710 µm)
2 0.183 0.8600 0.2486
3 0.183 0.8538 0.2470
Mean 0.1823 0.8579 0.2459
3 Fairly coarse 1 0.183 0.8490 0.2426
(≥500µm)
2 0.181 0.8468 0.2374
3 0.180 0.8513 0.2372
Mean 0.1813 0.8490 0.2391
4 Fine 1 0.171 0.8350 0.2362
(≥300 µm)
2 0.169 0.8400 0.2315
3 0.171 0.8420 0.2355
Mean 0.1703 0.8385 0.2344

39
40
The percent porosity of the sawdust sample is shown in Figure 4.1 as it relates to the size

distribution. The results indicated a percent porosity of 98.02±1.21, 97.73±0.82, 97.62±0.52 and

93.88±0.41 for the varying particle sizes of Very coarse (≥1 mm), Coarse (≥710 µm), Fairly

coarse (≥500 µm) and Fine (≥300 µm) respectively. As a result, a mean percent porosity of

86.04±0.03 was determined.

41
 0.92

0.9

0.88
Porosity

0.86

0.84

0.82

0.8
Very coarse Coarse Fairly coarse Fine

Particle size

Figure 4.1: Porosity versus Particle size

42
The sawdust Particle density in grams per cubic metre was determined as 0.2344±0.0055,

0.2391±0.0031, 0.2459±0.0034 and 0.2504±0.0007 for the Very coarse, Coarse, Fairly coarse

and Fine particle sizes respectively as depicted in Figure 4.2. The mean Particle density was

determined at 0.2425±0.0071.

43
 0.26

0.25

0.25
Particle density

0.24

0.24

0.23

0.23
Very coarse Coarse Fairly coarse Fine
Particle size

Figure 4.2: Effect of Particle size on particle density

44
4.1.2 Determination of Extractives

Water Soluble Extractives

The result for the determination of Water soluble extractives in the sawdust sample reveals a

weight of 1.705 grams. Its weight percent was found to be 5.442% of the 31 grams initial sample

weight as depicted in Figure 4.3.

Ethanol Soluble Extractives

Figure 4.3 shows the result for the Ethanol soluble Extractives. These Extractives account for

1.585 grams of the sawdust sample weight of 31 grams. It was found to occupy a weight percent

of 5.113%.

45
 30

25

20
Weight in grams

15

10

0
Water Soluble Extractives Ethanol Soluble Extractives Extractives-free Sample

Sawdust Constituents

Figure 4.3: Showing the weight composition of the water and ethanol soluble
extractives in relation to the sample weight

46
4.2 Oven Dry Weight

The Oven Dry Weight (ODW) for the proximate components was found to be 1.984 grams,

1.981 grams and 1.982 grams with a Total mean ODW of 1.982±0.001 grams. The percent Total

solid parameter was found as 99.35, 99.65 and 99.5 for the varying Oven dry weights

represented.

4.3 Sieve Analysis and Lignin Content Quantification

4.3.1 Sieve Analysis of Pre-treated Sawdust sample

Particle size distribution was performed after the pre-treatment (Removal of the Water soluble

extractives and Ethanol soluble extractives) of the sawdust sample. It was found to be 6.430,

5.045, 5.495, 1.900, 3.025, and 5.260 for ≥1mm, ≥710µm, ≥500µm, ≥425µm, ≥300µm and >300

µm particle sizes respectively. Figure 4.4 depicts the particle size distribution for the extractive-

free sample used for the acid insoluble residue determination

47
 7

5
Sawdust weight, g

0
1mm 710µm 500µm 425µm 300µm Fine dust

Figure 4.4: Particle Size Distribution of the Pre-treated sawdust

48
4.3.2 Lignin Content

Figure 4.5 show the effect of particle size on the extent of acid insoluble residue (Klason Lignin)

obtained. The optimum acid insoluble residue was obtained for the ≥1 mm particle size of

sawdust biomass. The results obtained were 0.2267±0.0080, 0.1813±0.0015, 0.1823±0.0012 and

0.1703±0.0012 for the Very coarse, Coarse, Fairly coarse and Fine particle sizes respectively.

49
 0.25

0.2
Percent weight of lignin, wt%

0.15

0.1

0.05

0
Very coarse Coarse Fairly coarse Fine

Particle size

Figure 4.5: Weight of Acid insoluble residue obtained versus Particle sizes

50
4.4 Acid Insoluble Residue (Klason Lignin) Histochemical tests

4.4.1 Toluidine Blue Test

Figure 4.6 shows a positive result for the Toluidine Blue test. As depicted, the dye had changed

colour from blue to pink after contacting it with 4 drops of the dye.

51
Figure 4.6: Colour change after toluidine Blue histochemical test

52
4.4.2 Phloroglucinol – HCL test

Figure 4.7 depicts a positive Phloroglucinol – Hydrochloric acid test. As was observed, a red

colouration soon appeared after the introduction of 2 drops of 2% (w/v) Phloroglucinol – HCL

solution. Figure 4.7 (a) shows the Acid Insoluble Residue just 3 – 5 seconds after introducing

the Phloroglucinol – HCl solution. It can be observed that a red colouration builds around the

sample as the solution disperses. Figure 4.7 (b) shows the red colour spread 10 seconds after the

staining and Figure 4.7 (c) shows the sample 20 – 25 seconds after the staining. It was observed

that the red colouration fades with time.

53
 Figure 4.7(a) Figure 4.7(b)

Figure 4.7(c)

Figure 4.7: Showing the red colour formation during the Phloroglucinol – HCl
histochemical test

54
4.5 Discussion

4.5.1 Effects of Proximate Components

The Ash which is the non-combustible constituent of the sawdust sample was found to be 0.01

grams (0.47 percent of the 2g used for the analysis). Ash is an impurity that will not burn, so

biomass with low Ash content are better suited for any kind of pyrolytic action than biomasses

with higher Ash content. According to Joseph et al., (2003), higher ash content in a fuel usually

leads to higher dust emissions and affects the combustion volume and efficiency. According to

Kim et al., (2001), the ash content of any biomass has a significant influence on the heat transfer

to the surface of the biomass as well as the diffusion of oxygen to the biomass surface during

combustion. Loo and Koppejan (2008) also reported, that the higher ash content in a fuel, the

lower is its calorific value which determines the energy content of the fuel. Since Lignin is richer

in carbon compared to cellulose and hemicellulose (Markus, 2006), then this implies that the

more Lignin content obtained, the higher the calorific value and the lesser the ash content of the

sample.

From Table 4.1, it can be deduced that the percent moisture content of the sample was

9.65±0.13. The Moisture content determines the total energy that is needed to bring the biomass

up to its pyrolytic temperature during pyrolysis. According to Joseph et al., (2003), it affects the

internal temperature within the biomass. The higher the moisture content of the biomass, the

more prolonged the pyrolytic process due to endothermic evaporation within the biomass, which

will consequently prolong its residence time within the reactor.

The Percent Volatile Matter (PVM) was obtained at 99.35±0.11. According to Joseph et al.,

(2003), biomass generally contains a high volatile matter content. This makes the biomass

sample, a highly reactive fuel. Biomass with high PVM give a faster combustion rate in the

55
devolatisation phase during pyrolysis. Biomasses with low PVM, have been reported to result in

smouldering which leads to a significant amount of smoke and release of toxic gases. However,

for the biomass sample at a PVM of 99.35±0.11, it is an indication of ease in ignition during

pyrolysis. It also indicates that during combustion, most of the sawdust biomass will volatise and

burn as gas.

It can be deduced from Figure 4.1, that the Percent Fixed Carbon (PFC) is 0.18±0.03. Biomasses

are rich in Carbon and Hydrogen. These elements represent the main heat producing elements.

The PFC represents the percent amount of carbon available for char combustion during

pyrolysis.

According to Table 4.1, the Percent Organic Content (POC) was obtained as 0.136±0.03. The

organic content of the biomass influences many of its physical, chemical and biological

properties. It affects its moisture holding capacity, shear strength, water and air infiltration rates

(Joseph et al., 2003). The higher the organic content of the biomass, the higher its moisture

retention.

The Percent Porosity was obtained as 86.04±0.03. Figure 4.2 shows the percent Porosity of the

sawdust sample as it relates to size distribution. It was observed that the percent porosity of the

biomass sample increases as particle size increases. The porosity of the sample is the measure of

the void spaces within the sawdust grains. It can also be inferred that the larger its porosity, the

better the diffusion rates of extractive fluids. Consequently, it was observed that the Lignin

content yield was more in the sample with a very coarse particle size compared to others. And

this yield decreased with a decrease in particle size.

56
The Particle Density (PD) was reported as 0.24±0.01 grams per milliliter (g/mL). It can be

inferred from Figure 4.3 that the Particle Density of the sample increases with decrease in

particle size.

4.5.2 Influence of Oven dry weight on proximate components of biomass

To get accurate PMC values, considerable care was taken in proper weighing of sections, in

proper operation of the oven, and in careful attention to calculating the ODW. The ODW was

used to determine the PMC of 2 grams each for nine experimental runs. It was observed that the

higher the ODW of the woody biomass the lesser the moisture content. Consequently, this affects

the Heating value of the wood specie as this has a direct relationship with the moisture content

prevalent in the wood. As lower moisture content will foster a higher heating value.

4.5.3 Effects of Acid Hydrolysis on biomass

Figure 4.8 shows the gradation formed after the Acid hydrolysis of the sample. The biomass

sample was hydrolysed using 72% sulphuric acid and 28% water for 6 hours. The hydrolysis

with the acid aimed at dissolving all polysaccharides within the sample leaving just lignin, as the

lignin condenses to become practically water insoluble. The acid cleaves to the glycosidic

linkages in the carbohydrates forming individual sugars. Consequently, the sugars dissolve in

Water after its dilution with 560 ml of water as the lignin does not and settles. It was also

observed that fragments of lignin were visibly seen afloat. As reported by Anilio et al.,(2000),

the hydrolysis with dilute sulphuric acid produced acid hydrolyzates with lower contents of

inhibiting substances like Furfural, Acetic acid etc.

It was observed that the Wood-Acid hydrolysis produced a liquid fraction and a solid precipitate

mainly composed of Lignin and cellulose. The liquid fraction as suggested by Anilio et al.,

57
(2000), contains sugars (Xylose, Glucose, Mannose, Galactose, Arabinose and Ramnose) and

other products (Furfural, Hydroxymethylfurfural and Phenolics).

Dilute sulphuric acid

solution

Acid Insoluble contents

Figure 4.8: Showing the gradation inside the dilute acid during Acid hydrolysis

58
4.5.4 Influence of Particle size on Lignin content yield

It can be inferred from Figure 4.8, that the larger the sample particle sizes the higher the lignin

content.

4.5.5 Influence of Extractives

From Figure 4.5, it can be inferred that there exist a direct relationship between the extractives

composition and the total sawdust weight. It was observed that an increase in extractives weight

would mean a decrease in sawdust weight. Invariably, this implies a decrease in Lignin content.

As suggested by ASTM E1690 (2003), these extractives if not removed could interfer with the

Lignin quantification resulting in error readings.

Additionally, the extractives content of biomasses are combustible matter. According to ASTM

E1690 (2003), these extractives consist of Starch, Colouring matter, Gums, Tannins, Sugars etc.

Consequently, the more extractives a biomass contains the higher its heating value.

4.5.6 Colour change during Histochemical Test for Lignin

Figure 4.6 indicates the Toluidine Blue test for Lignin in the Acid Insoluble Residue obtained

from the biomass acid hydrolysis. The illustration clearly depicts the colour change from blue to

pink. This colour change is as a result of certain functional groups that make up the lignin within

the test sample.

Figure 4.7 depicts the colour change during Phloroglucinol – HCl test. It clearly indicates a red

colouration formation round about the test sample. A very sharp violet colouration was also

observed on the certain parts of the particulate test sample. According to Jeffrey (1997), highly

purified coniferyl and syringyl alcohols both yield red chromophores. The chromophore

responsible for this colour change has been widely reported to arise from the acid-catalysed

59
condensation of phloroglucinol with the coniferaldehyde units in Lignin. The syringaldehyde

units have also been reported to yield a red chromophore. The intensity of the red colour gives an

indication of the amount of Lignin present.

60
 CHAPTER FIVE

CONCLUSION AND RECOMMENDATION

5.1 Conclusion

Based on the results of this study, the following conclusions can be drawn;

The proximate components for ash content, moisture content, volatile matter, fixed carbon and

organic content for hardcross hardwood were obtained as 0.47±0.10, 9.65±0.13, 99.34±0.11,

0.18±0.03 and 0.136±0.03 respectively.

The biomass porosity and particle density was obtained as 86.04±0.03 and 0.24±0.01

respectively. The extent of extractives either water soluble or ethanol soluble affects ultimately

the weight of the biomass and consequently, the estimation of its lignocellulosic constituents.

The lignin content of the hardcross hardwood is estimated at a mean value of 0.1902±0.0250

across the lignin content of its particle sizes of 1 mm, 0.71 mm, 0.50 mm and 0.30 mm.

There also seem to be a correlation between the porosity, particle density and the extent of lignin

yield. As reported by Ronald et al. (1994), there is an existent relation between the void spaces,

particle structure and lignin content yield. Nonetheless, it can be inferred from this study that the

extent of lignin yield is dependent on the porosity, particle density and ultimately, the type of

extraction procedure employed.

5.2 Recommendation

Since, lignin content varies over the extraction technique employed. It is recommended that

further research be done on lignin content estimation using other techniques. Consequently, a

comparison could be made and an argument raised.

61
Further research should also be conducted on other lignocellulosic constituents to see if this

relationship is existent.

1.5 Contribution to knowledge

This study has greatly advanced the need to understand the compositional analysis of woody

biomass, as the trend to harness renewable sources like woody biomass for energy matter is

growing.

62
 REFERENCES

A. Sluiter, B. H. (2008, 3 21). Determination of Ash in Biomass.

Anchukaitis K. J., E. M. (2008). Consequences of a rapid cellulose extraction technique for
oxygen isotope and radiocarbon analyses. Analytical Chemistry , 80 (6), 2035–41.
Anilio C., J. M. (2000). Wood Hydrolysis and Hydrolysate Detoxification for Subsequent Xylitol
Production. Journal of Chemical Engineering Technology , 23, 1013-1020.
Anu M. (1997). Introduction to modelling and Simulation. U.S.A: Department of Systems
Science and Industrial Engineering Binghamton, NY.
ASTM D1106-96. (2007). Standard Test Method forAcid-Insoluble Lignin in Wood. West
Conshohocken, PA, USA.
ASTM D1348−94. (2008). Standard Test Methods for Moisture in Cellulose. 100 Barr Harbor
Drive, West Conshohocken, PO Box C700, PA, USA: ASTM International.
ASTM D2974. (2002). Standard Test Methods for Moisture, Ash, and Organic Matter of Peat
and Organic Soils. ASTM Journal of Standards .
ASTM D346 D346M-11. (2011). Standard Practice for Collection and Preparation of Coke
Samples for Laboratory Analysis. West Conshohocken, PA, USA, www.astm.org: ASTM
International.
ASTM E1690. (2003). Determination of Ethanol Extractives in Biomass. Annual book of ASTM
Standards , 11.05 . Philadelphia, PA, USA: American Society for Testing and Materials.
ASTM. (2009). Standard Test Method for Determination of Acid-Insoluble Residue in Biomass.
ASTM E1721 − 01 .
Boerjan W., R. B. (2003). Lignin biosynthesis. Ann. Rev. Plant Biol. , 54 (1), 519–549.
Boman C., N. A. (2004). Energy and Fuels , 18, 338-348.
Boone R.S. and Wengert E.M. (1998). Guide for Using the Oven-Dry Method for. Forestry
Facts . USA: College of Agriculture and Life Sciences, University of Wiscousin-
Madison.
Bridgwater A.V. (2003). Chem. Eng. J. , 91 (87).
Celignis Limited. (2015). Relevant Chemistry. Retrieved from chemistry.php.htm
Chaney J. (2010). Combustion characteristics of biomass briquettes. University of Nottingham.
Chaplin M. (2014). Cellulose - Wikipedia,the free encyclopedia. Retrieved December 14, 2014,
from www.wikipedia.com/cellulose.
Darnoko D. and Cheryan M. Kinetics of palm oil transesterification in a batch reactor. Journal of
the American Oil Chemists Society , 77 (12), 1263-1267.

63
Davin L.B. and Lewis N.G. (2005). Lignin primary structures and dirigent sites. Current
Opinion in Biotechnology , 16 (4), 407–415.
Dey P.M. and Brinson, K. (1984). Advances in Carbohydrate Chemistry and Biochemistry.
Dinesh M., C. U. (2006). Pyrolysis of Wood/Biomass for Bio-oil: A Critical Review. Energy
and fuels , 20, 848-889.
EUROPA European Commission. (n.d.). Retrieved from
http://europa.eu.int/comm/research/energy/nn/nn_rt_bm/article_1111_en
Fiebach K. and Grimm D. (2000). Resins, Natural. Ullmann's Encyclopedia of Industrial
Chemistry.
Forest products laboratory. (1987). Wood as an engineering material. In Agicultural Handbook
(pp. 4-15,3-12). Washington DC: US department of agriculture.
Gellerstedt G., H. M. (2009). Wood chemistry and biotechnology . Göttingen: Hubert & Co.
Group, W. B. (Director). (2013). What is Energy Poverty? [Motion Picture].
Holt-Gimenez and Eric M. (2007). Biofuels: Myths of the Agrofuels Transition. Oakland,
Carlifornia, United States of America: Institute for Food and Development Policy.
Horisawa P. (1999). Determination of wood biomass porosity.
Hu G., H. J. (2008). Feadstock pretreatment strategies for producing ethanol from wood, bark,
and forest residues. Bioresources , 3 (1), 270-294.
Jaya S. T., S. S. (2011). A Review on Biomass classification and composition, co-firing issues
and pretreatment methods. 2011 ASABE Annual International Presentation.
Jean-Pierre B. (1996). Mémotech Bois et Matériaux Associés. Paris, France.
Jeffrey D. (1997). Lignin Analysis. Mississippi, USA: ResearchGate.
John M. H. (1966). Lignin production and detection in wood. U.S. Forest Service Research.
Joseph O. A., F. K. (2003). Physico-chemical characteristics and market potential of sawdust
charcoal briquette. Journal of Science and Technology .
Jung, S. K. (2008). Production of bio-oil from rice straw and bamboo sawdust under various
reaction conditions in a fast pyrolysis plant equipped with a fluidized bed and a char
separation system. J. Anal. Appl. Pyrolysis , 82, 240-245.
Kersten, S. W. (2005). Biomass pyrolysis in a fluidized bed reactor: Part 1: Literature review and
model simulations. Ind. Eng. Chem. Res., 44, p. 8773.
Kim H. J., L. G. (2001). Modelling Combustion Characteristics of Bio-coal briquettes. Journal
of Energy resources Technology , 123, 27-31.

64
Kittiphop P. and Tharapong V. (2012). Received Oil yields and Chemical compounds of Mixed
grass synthesized by pyrolysis process. IOSR Journal of Applied Chemistry , 3 (2), 14-20.
Klemm D., H. B.-P. (2005). Cellulose: Fascinating Biopolymer and Sustainable Raw Material.
Angew. Chem. Int. Ed. , 44 (22).
Komers K. (2005). Kinetics and mechanism of the KOH - catalyzed methanolysis of rapeseed oil
for biodiesel production. European Journal of Lipid Science and Technology , 104 (11),
728-737.
Kuye A. and Okorie O. (2014). Pretreatment of rubber tapping and achi mkpuru hardwood
sawdust for bio-oil production: drying and particle size characteristics at different
temperatures.
Kuye A., I. I. (2012). Pre-treatment of Danta and Hard cross hardwood sawdust for Bio-oil
production: Drying and particle size characteristics at different temperatures.
Lavarack B. P., G. G. (2002). The acid hydrolysis of sugarcane bagasse hemicellulose to produce
xylose, arabinose, glucose and other products. Biomass and Bioenergy , 23, 367-380.
Loo S. V., a. K. (2008). The Handbook of Biomass combustion and Co-firing. London:
Earthscan.
Markus H. (2006). Wood biomass as a fuel. Intelligent - Energy Europe Programme .
Mohammed I. J., M. G. (2012). Biofuels Production through Biomass Pyrolysis-A Technological
review. Energies , 4952-5001.
Mojmír N., L. W. (2010). Alternative methods for cellulose preparation for AMS measurement.
Proceedings of the 20th International Radiocarbon Conference,. 52, pp. 1358–1370.
Arizona: University of Arizona.
Mosier N., W. C. (2005). Features of promising technologies for pretreatment of lignocellulosic
biomass. Bioresource Technology , 96, 673-686.
NREL 42619. (2008). Standard Method for the Determination of Extractives in Biomass.
National Renewable Energy Laboratory. NREL BAT Team Laboratory Analytical
Procedure.
Nwakaire J. N., O. O. (2011). Determination of Physio-chemical Properties of Oleander Seed Oil
for Biodiesel Production. Renewable Energy For National Industrial Growth, (pp. 32-
35).
Nwakaire J.N and Ohagwu, C. (2009). Design, Construction and Test-running of a biodiesel
pilot plant. CTA/ATPS/AGRA/FARA/NEPAD/RUFORUM.
Okorie O. (2014). Pre-Treatment of Rubber Tapping and Achi Mpuru Hardwood sawdust
forBio-Oil Production: Drying and Particle size Characteristics at Different
Temperatures. Port Harcourt, Nigeria: PTDF.

65
Peterson C.L. and Wilkerson D.F. (2002). Continuous Flow Biodiesel Production. Applied
Engineering in Agriculture , 18 (1), 1-5.
Petura R. C. (1979). Thermodynamic data for waste incineration. American society of
mechanical engineers.
Ragland K. W. and Aerts D.J. (1991). Properties of Wood for Combustion Analysis.
Bioresource Technology , 37, 161-168.
Rizki M., T. Y. (2010). Scrutiny of Physical Properties of sawdust from Tropical Commercial
Wood species: Effects of different Mills and sawdust particle size. Journal of Forestry
research , 7 (1), 20-32.
Ronald D. H. (1994). A comparison of the Insoluble residues produced by the Klason lignin and
Acid Detergent lignin procedures. Journal of Science, Food and Agriculture , 65, 51-58.
Ronald H. and Romualdo S. F. (2005). Can Lignin be accurately measured?
Sadaka, S. (2010). Pyrolysis. Nevada, Iowa, United States of America: Iowa State university.
Samuel P. H. (2014). Lignocellulosic feedstocks: research progress and challenges in optimising
biomass quality and yield. Frontiers E-books.
Sandra, D. (2010). Production and characterization of bio oil from wood. Pahang, Malaysia.
Soltes E. and Elder T. (1981). Pyrolysis. In Organic Chemicals from Biomass. Florida: Goldstein
IS. (ed.) CRC Press.
Tillman D.A, R. A. (1981). In Wood combustion: Principles, processes and economics (p. 43).
Academic press.
Turner, T. L. (2005). Modeling and Simulation of Reaction Kinetics for Biodiesel Production.
Updegraff D.M. (1969). Semi-micro determination of cellulose in biological materials.
Analytical Biochemistry , 32 (3), 420–424.
Weatherwax R. H and Tarkow H. (1987). Density of wood substance-Importance of penetration
and absorption compression of displacement fluid. Forest products journal , 18 (7), 44-6.
Woodgas. (2007). www.woodgas.com/proximat.htm. Retrieved from Woodgas.
Woody Biomass Properties. (2015). Retrieved from
http://www.extension.org/pages/26517/woody-biomass-properties#.VMkqUWdsxQA
Xiaoli G., X. M. (2013). Pyrolysis of poplar wood sawdust by TG-FTIR and Py–GC/MS.
Journal of Analytical pyrolysis , 16–23.
Yang B. and Wyman C.E. (2008). Pretreatment: the key to unlocking low-cost cellulosic ethanol.
Biofuels Bioprod. Biorefining , 2, 26–40.

66
Yulia G. (2011). Extraction of Hemicelluloses by Acid Catalyst Hydrolysis. Imatra: Saimaa
University of Applied Sciences.
Zhang M. M., K. V. (2005). Thermodynamic properties of 1-butyl-3-methylimidazolium
chloride (C4mim[CI]) ionic liquid. Journal of Phase Equilibria and Diffusion , 26 (2),
124–30.

67
 APPENDIX A

BUFFERS

A.1 Phloroglucinol-HCl solution

A buffer solution of 2 w/v% phloroglucinol solution represents a mixture of 0.05 grams of

phloroglucinol in 25 ml of ethanol. Two (2) drops of HCl was then dissolved in five (5) drops of

the phloroglucinol-ethanol solution.

68
 APPENDIX B

STATISTICAL RESULTS

B1: Proximate Components

Proximate Component Statistical data

Experiment Standard Average wt%
I deviation
PAC 0.32 0.03 0.35
0.38
0.35
Experiment II
0.58 0.02 0.56
0.55
0.56
Experiment III
0.50 0.02 0.50
0.49
0.52
PMC Experiment I
9.82 0.18 9.69
9.48
9.77
Experiment II
9.75 0.12 9.61
9.53
9.56
Experiment III
9.78 0.11 9.66
9.64
9.56
PVM Experiment I
99.53 0.05 99.48
99.43
99.47
Experiment II
99.19 0.04 99.24
99.25
99.27

69
 Experiment III
99.35 0.02 99.33
99.31
99.32
PFC Experiment I
0.15 0.02 0.17
0.19
0.18
Experiment II
0.23 0.03 0.20
0.20
0.17
Experiment III
0.15 0.03 0.17
0.20
0.16
POC Experiment I
0.09 0.01 0.10
0.11
0.10
Experiment II
0.16 0.00 0.16
0.16
0.16
Experiment III
0.15 0.01 0.15
0.14
0.15

B2: Lignin content, Porosity and Particle density

S/N Particle Experiments Percent weight Porosity, % Particle Density,

Size of Acid Insoluble grams per ml
residue, %wt (g/ml)
per gram
1 ≥1 mm 1 0.236 0.9712 0.2505
2 0.221 0.9756 0.2510
3 0.223 0.9939 0.2497
Mean: 0.2267 0.9802 0.2504

Standard Dev: 0.0081 0.0120 0.0007

70
2 ≥710 µm 1 0.181 0.9865 0.2420
2 0.183 0.9740 0.2486
3 0.183 0.9712 0.2470
Mean: 0.1823 0.9772 0.2459

Standard Dev: 0.0012 0.0081 0.0034

3 ≥500 µm 1 0.183 0.9736 0.2426
2 0.181 0.9726 0.2374
3 0.180 0.9822 0.2372
Mean: 0.1813 0.9761 0.2391

Standard Dev: 0.0015 0.0053 0.0031

4 ≥300 µm 1 0.171 0.9430 0.2362
2 0.169 0.9385 0.2315
3 0.171 0.9348 0.2355
Mean: 0.1703 0.9388 0.2344

Standard Dev: 0.0012 0.0041 0.0025

71